onthe

*d
CAUSES
 onthe
*d
CAUSES
CONTROLoT
ACTI\ATED
SLUDGE
BULKING,
FOAMING,
*d OTHERSOTIDS
SEPARATION
PROBLEMS
Michael
DavidJenkins. G.Richard.
GlenT.Daigger

'l||
LEWISPUBLISHERS
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 Library of CongressCataloging-in-Publication Data

Manualon rhe.auss md conbl oI actimredsludgebulting,bming,6d otherlolids

sptratior prcblems/ by David Je.tins, Michael G. Ricndd, ed CI.n T. Daigger lrd ed

Rer ed. or: Manual on the causesand conlrol of &dvated sludsebulkine md foming.

lncludes bibliogiaphicol €feEnces od index.

ISBN 1-566t0-64?-5(alt. paper)
I Sewas€-Punicalion Activatedslu+e press. 2. Sludgebulkins. 3 Fl@cnlation.L
Richdd. Mich&l C. tI Daigger,Glen T. nI. Ientins, David, 1935' Manual or the causs
and control of activateddudge bulling dd foaning. rV T e.

TDt56.J,l6 2003
628.3'54-dc2l 2003051956

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 Dedication

To Samanthaand Maggie Jenkins,Jack Muren, Joan Richard,and Patty Daigger

t-
Preface
...But if I could st.rt all ovcf, gucssl'd still do what I do.
Wishire all dDsc old thingt ve.e new.
Merle Haggard,1937-
Activatedsludgeis the most $,idcl] uscdsecond:rrywaste-
rvater neatment proccss in thc world. Il consistsoi trvo
treatmentunits- thc acntnnr basin nnd the solids sepa
ration dcvicc - usuxlly r gruity sedinentation basin
called thc sccondar) .larilier Until about 20 years ago.
nrcs! invenigations of aclivxted dudge rvere concerned
with factorsaffectingpoll trtxnrremo\ al eticiency by pro-
cessesthat oc.ur in tho xcrationbasir even though mary
irlestigatorsand pr.tctitioncrsacknolvledgedthat the great
majority of cfllucnt qurlity problems were related !o rhe
in$ility of thc sccondrry clarilier Io eflicientty remove
the a.iivated sltrdgcbiomassfiom the treatedsastewater
T{) undcrst.tndthc bchnviorof aciivatedsludge in sol
ids separationproccsscs,il is necessaryto characterizethe
complcx commuDitythat makes up the actlvatedsludge
biomassruthcr than rcgard ir simply as susperdedsoljds
or volatilc suspcDdcdsolids.It is also important io under
stnnd that wrstcwalcr characterislicsard aerationbasin
environmcnrrlcondilions and coniigurationinfluencetle
way in $4richlhe aclivrlcd sludgesusperdedsoiidsbehave
nr a sccondarl clariflcr The design and operation ol an
a.tivated sludge slstcn to produce high quality effuen!
requiresthc intcgration of ieradon basin and secondary
This manualdcals with nll aspectsof activatedsludge
solidsscparationproblcns. ChapterI describesthe Daturc
of activatcdsludgc and its solids separationproblems: it
tr'lccsthcir o gins to tundamentalphysical.chemical.and
microbn)logical propcrtics of the acti\ated sludge. The
cuncnt undcrsrandingof activatedsludgebiollocculation
pn)ccsscsand thc cifccts of lilnmentousorganismson no.
structurcarc discusscd.
Chxptcr2 prcscntsmicroscopic,physical.andchemical
mcthdls ior activatcd sludge characterization and lirr the
diagnosis of solids scpffadon problems and iher causes.
This sccti(nris Ubcnlly illustrated\riih photomicrographs.
Dctailcd dcscriFn s and al1identification key fbr filanren
tous organismsarc prcscnled.The use of nole.uLar biolog
ical mcthodsis discusscdbnefly and sunmarJ lformation
on thc cuncnt status of lilamentous organism iderrification
i, f n\ c nr nl. V rrl -.J . n " b i n p l ),rn e ra n i l ),rs Jre ci \en.
Techniqucsfor differcntiatnlgbelween"biological-related'
and "processrelNled" separatior activated sludgc sotids
separationproblcmsarc discusscd.
Chaptcr 3 prcscntsan cxrcndcddiscussionof how a
microscopicclxluation oiactivlled sludgecan be usedto
heb evaluatcsdids separrrionproblclns.Thc usesof both
floc and lilamentousoreanismobscrvarbnsare described.
Chapter,l is { discussionon how to rcmcdy and pre-
vent a.tivated sludgesolidsscparalbn problems.The the-
ory of rctivatcd sludgc bchaviorin secondaryclarjiiers is
discusscdrnd is uscd10dclclop methodsof clarifief and
acralion basin mrnxgcmenr lbr combatting bulking
sludgc.SccoDdxr)clarificr opcraringdiagramsfbr severai
scttUngtcslmcthodsarc given.The useofpolyners. chlo
rinc. hydrogcn peroxidc. .tnd ozone lbr bulking sludge
conlrol is prcscnled.logerherwith casehistoriesfrom full
scalc lrcxtnenl planrs.The resolutionof specificbulking
problens due lo macro- ard micro-rutrient defciency.
sullidc. low dissoh€d oxygen concentation. and readily
metabolizeablesubshies is given in detail: the effecr of
rcralion basin conJigurationon activatedsludge settl"rg
xnd the design and use of selectorsfbr bulkirg control is
colcrcd in dcpth; illustralive examples from full scale
lrcatmcnt plants arc presented.Methods for dealing with
dispcrscdg$wth and viscousbulking are discussed.The
usc of licld and laborabry testingmethodsibf dilTerenti
xting belween "biological" and "process- problems is
illuslrated wiih case histodes from full s.ale trcatmcnt
Chapter5 discussesihe causesand control ofacti\ated
sludgefbaning. The roles of sudactants.
'nicutrgamsms
suchas nocardiofbrms.andM,'rothri:t parrk ella.re ort-
lined: the eftbctsofphysical detajl! of thc acfutionbasin.
secondaryclailier, and in-plant recycleson n)rrn reten-
tion and fbam recycling are discussed.Tbc control of
nocardiolbnn organisns by using lo\ MCRT and various
types of selectorsystemsis discnssedxnd illustratedwith
caschistories.Nocardiotbrmfoaming.ontrol by calionic
polymer addition.surfacechlorine mist sprayapplicaiion.
seleclive surface sludge $asting. and automaric MCRI
control is presented. The impacl of nocardtufbrm organ'
ism fbaming on anaerobic digestion is discussed.The
chaptercloses a discussionof thc causesand contlol
'vrrh
merhods ibr bulking and foaming causcd tJy Mictothrix
The manualcontainsan exrensivcbibliographyon all
aspec$of activatedsludge solids separllion problems.
This manual is designed to be usciul to the wide
vadcty of prolessionalsNho design.managc,monltor.and
operatetheactivatedsludg€proccss.Materialin this text designandop€.ation.Its pr€viow €ditionshavebecnused
is usefulfor labomtorypersonnel,
alesign
engine€rs, and widely in labontory couses on activatedsludgecharac-
treatrnen!plantoperaton,Th€manualservesasa usefrrl lerization and solids seDarationDroblerns.
texton activatedsludgemicmbiologyandtreatnentplant

L-

t-
 Acknowledgments
This rnanual was rnade possible by the work and slpport
of many individuals and organizations. Our sinc€re than*s
and gralitude are due 1() them. The iirst ediiion of the
manual was rnade possible by grants liom the Water
ResearchCommission of the Republic of South Ahica
and the U.S. Environmentalhotection Agency.
Much of the research that laid the foundalion for this
mrnual was conducted over the lasi three and a half
decadesby a dedicatedand imaginativegroup of graduate
students and post doctoral researchen at ihe Unive$ity of
Califomia at Berkeley Although their names appear in the
bibliography,they, of all people,deserveindividual men-
tion. The] are Mesul Sezgin, Denny Parke! Jonathan
Palm, Tony Lau. Peter Stron, Sang Eun Lee, Ben Koop-
t_ man, Oliver Hao, Michael Richard,J.B. Neethling, Greg
Shimizu, Kay Johnson, Andrd van Niekerk, Benardo
Vega-Rodriguez,H3ro Bode, Y-J. Sbao,Paul Pitt, Daniel
Cha, Daniel Mamais. Linda Blackall, Valier Tandoi,Mark
Hemandez, Cho Fei Ho, Jean Weber Krishna Pagilla,
Andy Schule! Willie Harpe! Abe Fainsod,Trina McMa-
hon, SuzanYlmaz, B- Narayanan,and Carlos De L€6n.
We owe a special thank you to the wasiewater treat-
ment plant personnel who have tried out our ideas in
practice.Specialrecognirionis due to thc peoplewho took
special risks in "doing it first." Thesc are Russ Edwards,
Albany, Georgia;Bob Beebe,San Jose/SantaClara, Cal-
ifomia; RandyGray,StrohBrewingCo., Longview.Texasi
Bill Keaney,North Sltn Mateo Sanitation Distrjct. Cali
tbmia; MiLe Wheelcr,Hamilton, OHi Millard H. Robbins,
Upper Occoquan Scwxgc Authority. Cenfeville. VAi
Michael Read and Richard Stillwell. ClackamasService
Districl, Oregon; Gary Vaughn and Billy Ammons, Fay-
elteville, Arkansas: Dale Richwine and Carlo Spani.
Unned SeweragcAgency.OregoniJohn Reid. StoneCon,
tainff Corporalion.Ontonagon,Michigani ToddCockburn
and JonathonLoiacono, San Francisco.Califomia; and
Wendell Kido, Sacrarncnto.Califomia.
We would also likc to acknowledgethe contributions
of our colleagueswho frecly and openly discussedour
ideas and often providcd us with fi€sh ;nsiehts.Thesc
include Onie Albertson,Denny Pxrker.Tom Wilson. Wcs
Eckenfblder,David Stensel,Bob Okcy. John NovJ<, Jay
Keasling, Merv Goronsczy, Alex Ekslet and Joh Kang.
The photomicrographswere tal€n by Michael Rich-
ard. SuzanYilmaz and SarahMuren producedthe ngures.
Sarah Muren and Joan Jenkins prepared the manuscripl-
JoanJenkinsvedfied the cilations.The palience.skill and
hard work of all thcsc people arc ackno$ledged.
David Jenkins, Berkeley, CA
Michael Richard,Fo Couins,CO
Glen Daigger Denver, CO
Authors
David Jenkins is a professorin
the graduateschooland profes
sor emeritusof envi.onmental
engineering at theUniversityof
Califomia arBerkeley.He cane
to the United Statesfrom Great
Britainaftereaminga B.Sc.ill
applied biochemistryfrom Bir-
mingham Universiryand a
Ph.D.in publichealthengineer
ing from the Universityof
Durham.King'sCollege.Professor Jerktus'researchand
professional practiceis in the generalareasof biological
wastervatertreatmentaDdwaterandlvaste$aterchemistry.
He is ihe authorof over 250 publications. His research
workandprotessional contributions havebeenrecognized
by arvardsfrcln theWaterEnvironment Federation (Eddy,
Gasgoine. Camp,andFairMedals),theAmericanSociety
of Civil Engineers(FreezeMedal).lhe Associationof
Environmental sngineefiDg andScienceProfessors (Out-
srandingPublicatjon Alvard).andthe Inremational Watel
Associalion(San JenkinsMedal and the Ardern and
t-ockett Awafd). He is an honorary life memberof the
WaterUnvironmentFederationard the lntemationalWater
Association and a memberof the NationalAcademyof
Engineering.The resuhsof his researchon activated
iludge solids separationproblem! have been applied
worldrvideto impro\'ethe designandoperationof biolog-
ical wastewarertreatmentplants.
MichaelG. Richard is a senior
ervironmental scientist with
SedrBro\\nin FonCollins.Col-
orado.His educationincludesan
A.B. in field biologyandecol-
ogy and an M.S. and Ph.D.in
environmentalhealthsciences
fiom the Univenity of CalifoF
nia at Be*eley. Prior to his cur'
rcnt positlon.he lvas a rcsearch
specialistin wastelvaterteat-
mert microbiology at Berkeleyand an assistantprofessor
of environmentalhealth at ColoradoStateUniversity.He
is the authorof aboui i00 papersandte.hnical reponsand
theauthoror coau*nr of live bookson wastewater aeahent
microbioiog}' He has given more than 150 Fesentaiions
at meetingsnaiionwide. He is lhe recipient of the Eddy
Medal liom the Wxtcr Environment Federation, the Russell
BlosserAward horn thc TcchnicalAssociationof the Pulp
and Paper lndustll,. and the Douglas Jones Award from
the Pulp and PaperTechnicalAssociationof Camda. He
was named the Environmental Educator of the Year in
1999by the NationalEnvirormentalTrainins Association.
Dr Richard has bccn involved nr the diagnosisand cor-
rection of wastewxtcrtreatmenl microbiology problems
lbr more than 300 cilies and 200 mdustrie!.Hehas ofiered
wastewaternicrobidogy trainnrg .ourses at over 125
iocalions tuoughout rhc U.S. and Canada.
{}l€n T. Daigger is a rcgistered
profcssn)nalengineerand is cur
rently sc.n)r vi€ presidentwith
CH2M HILL. wherc he reNes
as chief lechnology oflicer for
the lirm's rlatcr businesses. He
is also a tcchnical fellow in
waslewalcr lrcalment technol
ogy and as such he seNe! as
scnior consultanl and process
cnginccr on r wide variety of
municiFl and indusdxl wastcwatcr trcatmcnr dd reclama
tion projects.Dr Daiggeris the aulhoror coauthorof over
I 00 technicalpublicanons,sevemlmanualsthrl rrc widely
used in the wastewater professjon, and four b(x)ks. Hc wits
educatedar PurdueUniversitywhcrc hc camcd a B.S.C.D..
M.S.C.E..and Ph.D. in environmentalengineenng.He i! a
member of the Wa€r Environmenl Fcderalbn (wEFl.
American water works Associadon,American SNiety of
Civil Engineers, Iniemational Water Asnxiation. md the
Association of Environmenlal Engineerlng and Scienc€
Professors,and he is a diplomate of rhe American Academy
of Environmental Engineers (d{EE). He is a member of
the Nationai Academy of Engineering. Hc scrvcd as thc
chan of the WEF rask tbrce that preparcd the currcnl edition
of Manual of Practice 8, DesiKn ol Muntiry WastNdter
Trcatment PIu ts, ard chair of lhe board of ediloriai revie\v
ot Watu| Entionmtnt Resedf.n. He cunendy is chairof the
re.hnical Factice committee and research symposium of
the WEFTEC Prognm Commitee. He has received the
Gascoigne and Morgan medals liom WER and is the only
back-lo-back $inner of the Harrison Prcscoa Edd! a\\,:rd.
He has served as ihe Kappe lecturer for the AAEE, and he
is cunently a member of thc Walcr Environment Rcscffch
Foundation Resenrch Council.
 Listof lllustrations
ftcuRE1.1 Proposed
rolesof polrsaccharider
ard prcrcins(biopolyne6)anddilalcnt crtons i| biofloccutdLion

FIGURE 1.2 Phaseconlmsttnicroscopic appearance\ ol slrongactiEtedsludgelocs... ,.........................

...........................,..,..... l
FICURE,I.] E tc c t o f s h e .rr.1 eo n 1 4 0 0p n a . Li vared sl udgcfl ocs.... ...................... . . . . . . . . . . . . . . . . . . . . . 1
FIGURE 1.4 Phas.conlraslmicrognphsolelfectsof fillmentousore.nismson floc struclurcs...........................................,..,..4
FIGURE 1.5 P h a s e c o n tra s rmi c ro s c o p i .r ppearan.eol di spe.scdefow l h..................................... . . . . . . . . . . . . . . . . . . . . . . . . . . . .
FICURE 1.6 P h a s c o n tra smt i c ro Sra pohfsre l e sestxi ni ng of actnaadsl udge. ..........................
.. .............
.........
.. ...... . . . . . . . . . . 5
fIGURE1.7 E fti c to t c x @ c l l u l dl o l y s a c c h rri de
conl enl
on theS V I ol acti vared
sl udge........................
.. .. ............. . . . . . . . . . . . . . . . i
ftcuRE1.8
ftcuRf 1.9 Phasecontra\thicrcg.aphsot cilcctsof nlanenrds organism\on foc morphologyandserLleabiliry ... . .. .. ..6

FICURE 2.I S c h e m a ti c o u tl i n e o i n o c a rd i of(rmofgani smcouti ngnerhod..................................... . . . . . . . . . . . . . . . . . . . . . . . . . . . .

FIGURE 2.2 M o d e rn d n o c u l a rp h a s e c o n rr asl mi croscopccqui pped$i $di gnal crmc.a.................... . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
FIGURF 2.3 PhaseconlraslmicroFaphso
FIGURE 2.4
FIGURE 2.5
FIGURE 2.6 Pha\ccontrasrmicrogralhso
ftcuRE2.7 Ph a \ec o n t.a smt ' c ,o g rrl ho f fl o cwl tl poordi re(i ty..... ....................... ...... . . . . . . .22.....
ftcuRE2.8 Pha\econt.a\tmicDgraphsof nlamertabundrnce categorie\usingsubjecnve scorlrgstsren.................... .........2.,1
ftcuRE2.9 Phasecontn\t hic.osraphsof bmnchirgofnlamentousorgaiisnsiI acnvatedsludge.... .. .. .. ............ ..............27
rlcuRE2.10 Ph a s e c o l rn !tn i .ro g .a p h s o f1 j l !mcntshapes' ' ' ' ' ' ' ' .' ' ' ' ' ' ' ' ' ' .' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ,' ' ,' ' ,' ' ,' ' .' ' .,' ' ,' 2
flcuRE 2.11 P h a s e c o n tra s L mi .ro g ra p h s show i ngl ocrl ti otrsofnl amertousorgari sms................... . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
FIGURE 2.I2 Phasecontrastmiciotmphsshowingatt&hcdgrcrvthof b.cteriaon nhmentousorglnnms...............................29
FICURE 2.I3 P h a s ceo n tn snl i c ro € :ra pshhso w i ng appeaurccs of shcl l hs.... .....................,..,......,.............. .. .. .......................30
.
FIGURE2.I4 P h .s e c o n tra s l m i c ro 8 ra p h s ofnhmentousorgani smcc11shapes............................. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
FIGURE2.I5 P h a s c o n l ra smi sul furgranul c
l c ro g n p hosfi n ra cel l ul ar ...................... . . . . . . . . . . . . . . . . . . . . 32
FICURE2.16 D i rc c l i l l u m i n a ti o n d i c rc e r.p hsofP H A stai ni ng.P l l A sranul csaredarkbl ue/b1ack......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
FIGURE2.17 D i rc c ti l l u mi n a ti o n mi c ro g ra phsofGrxosLai nni gofl i l amcntousorgani sms................. . . . . . . . . . . . , . . , . . , . . . . . . . . . . .
ftcuRE2.18 D i re c ti l l u mi n a ti o n n i c fo e ra p hsofN el ssersl ai ni i gol nl anrentousorel ni sms................ . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
fICURE2.19 Phasecontc\t microgruphs of rcseltesand9onidia........... .. .....................................,. 15
ftcuRE2.20 Di.hotonousle! for idcnnficatio!ofilamertous orgxni$nsnr &dutcd sludge.................................................. l6
rlcuRE2.21 Phaseconh'asL fticsgnphs ot l^) Sphaerotilus kdidns.lb)'ltpe t7tJ1,(c) Haliscanenobddet bd^is... .. .....10
EICURE2.22 PhasecontiasL icmFrphs of: (a) Trpc 021N,(b) Type021Nlnh sulfu.granulcs,(c) Bessnttu.sp..

FIGURE ot: la) Thi.rhrLrt. (b) ThiothrirI ivith sulfurgiinules,(c) Thioth,& tt, afi
2.23 Ph.secontraslmicroeraphs

FICURE
2.24 Phasccont?srmicroSraphs ol (r) Type0914 (b) Tlpe 0914with sufur grdules.....
fIGURE
2.25 Phasecontrastnricroerallisof: (^) NolloLaiduliniola t, (b) Nosto.oi.ldlihiolall-(c) Nonooila

ftcuRE2.26 Phd\econt.a\tmicrcgraphs of: (a) Type0411.(b) Type0961,(c) Tytc 0092,.nd (d) Tlpe 0581 .......................42
FICURE2.27 P h a s e c o n tn \tmi c .o g ra p h s o r :(!)Tyl e0041,(b)Type067s.(.)Trpcl 85!.and(d)Tl l c0803.. . . . . . . . . . . . . . . . . . . . . . . 43
rlcuRE2.28 Phasecontnd mi.rog.aphsol: (.^)Mitothrix pdrti.dla. (b) Nocardn)tnm\.(c) Typc lE6l, and

FI6URE
2.29 Phasecontrastdicrogiaphsott (i) Fkribatter sp..h) Bd.iUt6 sp..t..) OyiUuhria V. (q,anophyccac),

FIGURE
2.]O 1, si/! idenlllicationof.irifying bacreriain adnaleddudgc:(a) slmultaneous
i, rir" hybndiraLnrr
wnh
Cyl labcllcdprcbeNmV.nd FLUOSlabelledpn'beNEU and(b) simultaneous d jt, identncarionoi
Nitosocaccrs habilis Nirlosld/a like bacreriaairer i' rn, hlbidiz.tion wirh FLUOS iabelledp.obe
N mV a n dC y 3l a b c l l c^td
d rc tES + N rspr1026a 18...........................................................
p .. ...... . ........................47
....
ftcuRE2.31 Phd\econtrastmicrcgraphs of conmonflaeellates and.nebae foundin activatcdsludgc..................... ............ad
fICURE2.32 Phasecontrastmicrugraphs of ilcc ud stalkedcillarcs..... .........................................
49
ftcuRt 2.33 Phase.ontm\tmic.og.aphs o
HCURE2.34
F I G URE2. 35 Phasecontnst microgcphs o
FI CURE2. 36
Ft c uRt 2. 37 Scheftaticdlagrumof highsolidslbamingtestappdaus 54
Ft c uRt 2. 38 Schematic diagramof Kemmerersamplerin use................ 55

FI CURE3. I
fI G URE] . 2 of SVI with nhmetrtlcnsthslor L4full scaleCalilorniaweltewalertreatmcDt
Varianons plants .. ..5E
FI G URE3. 3 ol subjective
RelanonshiD scoringol ilament abundancclid SVI asneasuredby indcpendenr obseryets. .. .5li
F I CURE3. 4 Comparisonof nlamenlcountlnd SVI ai the SanJosc/SanLa Claraplantin Calitbmia. . . . .. .. .59
T I G URE3. 5 of filamentcount
Rclarionships lo SVI for n61!!d second stageactivatedsludgeplantsal lhe Sar Jose/Santa

F I G URE3. 6 h lsn o n mi crobi!ani

Ph a s ce o n tra smt i c rc g ra p o ai cl es....... .. ...... 62
F I G URE3. 7 Phasecontrastmicrcgrathsof nonmicrobial padcles....... . . . .. .. .. . . 63
F I G URT3. 8 PhA* conlrastmicrcgrlphsof nonnicrobial pafticles......
F I CURE3. 9 P h a s e c o fl l ra s rmj c ro g ra p h s shoti rS morphobgi cal di versi l yofacl i vl tedsl udgefl @ s...... . .. 04.
sl udS f@
FtcuRt3.10 N e i s s epro s i ti vcce l l c l u m phs a c l i vatcd e s...... .......... . . . 0t )
f I G URE3. 11 PhasccontranmicoSraphsoil (a) yeist, (b) drProni./o biun P . l.) SpiriLluftsp and(d) spirochaeres . .. . 6?
f I G URE3. 12 Phasccontr$L microSlaphs of (r) lnicelhlar Chlorallasp dnd(b) lilanentous
U/o".ud sp . . . . . . 611

F I CURE4. 1 Schcmaric sludgeprcccs.........

diagramof the aciivarcd . . . . . . .. .. ?
FtcuRt4.2 Z o n cs c r1 l i i o8 f a c ti v a tesdl u d s e{ s tded so1i ds........... . . ... ..... 80
FrcuRt4.3
FlcuRt4.4
FI6URE4.5
FICURE4.6 t i x a mp l oe f s c c o n d acryl a ri fl el rh i c tcnl ngmal ysi s.... ..... ... ... 84
F I CURE4. 7
f I G URE4. 8
FtcunE
4.9 E ffe c to fu c u ti o n i c p o l y m€ ro n thcsvl ofabul ki ngsl udrg---.-----.-- ..... . 81i
FrcunE
4,10
HCURE 4,11 E x a mp l c s o i g o o d c h l o ri n c d o scpoi nr$forbul ki ngconl ro| .............. . 91
FtcuR[4,12 ChlorincresiduAlin RAS rs a iirnclionof initirl chlorinesPecies, reactiontimc.anil thc presence
oi

treatmcntpllnt anr'l(b) longaemtiontimc

4.I3 Chkrinationpafamcicfsii)r (d) lypicalmunjciPalwasLewd€r
FICURE

HCURE4.14 Ph a s ceo n trn smi r c ro g ra p sh hs o w i n proS

c rcssietfects
ve of chl ofi Mti onon l rntl ri r 1... . . ..... 95
FICURE GA w rstcw el cr
atthcA l bany,
4.I5 C o n rrool f b u l k i n gb y R A Sc h l o ri n rl nnl tredl ment pl atrr' . . ......... 96
-
FICURE
4.16 Useof tafgerSVI 10controlRAS chlorination dosefof bulkinScontroll|t theAlbnny,CA w6rtewaler

FrcuRt
4,r7 of RAS chlo.in0lioncftlctilsne$ whendosinechknnc to lhe mixcdliquorchunnelnnd
Comparison

F I CURE4. 18 SVI lnd chk ne doseto RAS at the SaoJose/S.nEClalawnterpoUrrioncontol llanl dunng$e l98l I)cdk

F I CURE4. I 9 WccklyalerngeSVI tiom 1979lo 1984ar $c StrohBrewingCo. actilatcdslDdgeplAnt,Longvicw,TX .. . 99

F I G URE4. 20 SVI andchlo.inedo* a1a wdnewarerrclivltcd $udgeplantar a plxsticslilctoryin March1984 . . .. .. . . 100
fIGURE 4.2'I EflluenlISS concenration andchlorincdoscat a waslewater
acrivalcd sludgeplanlnt r plasricstact(rv rn

.nd chlorjnedoserr a waslcrateraclivatedsludgeplanl!t d Pldsrics

T I G URE4. 22 EllluenlsolubleCOD concenfations

ftcuRt 4.23 Fled sysEmfor dosinghydr

ftcuRE4.24 SVI responsc 10hydrogenperoxidctrcdmentai the Petalunawslerpollulioncortrol Plantin Californir ' .. 102
HCURT 4.2s E fe c l o f o z o ser i ng..............
n a ti oonn a c ti v a tcsdl udge ...... .. , . . . 102
F I CURE4. 26 Monad model growtb cunes tbr ammonia-limitedgrcwth of Type 02lN rld a floc limet nohled from

F I CURE4. 27 Barchculiureammoniauplakeof rype02lN andan acdvlredsludSefl@ lbrmerpreliousiySrownunder

ammoni,limil ion andslbiectedto spitesin ammoniaconcentration. . .. .. .
........... 105
F I G URE4. 28 AmdoniAlimitedgowth conperilionbetween Type02lN (.) ald atractivared sludgefl( formcr(r) ii
chemosrat cullue Nith illemitrent (I) andcontinuods(C) NHr N lccdlng.C/N = 15with int rmiltent
feedingof 7591'of totalannonia ai 6 hourjnterlah....... I 05
fICURE 4.29 Effe.r of inillal mrerobic zoneon liminng intuenr COD/P ratio tbr a labontory scaleSBR actiured
106
HCURE4. 30 Efact of microNrrientaddltionon COD removalcfficicncyat. perochemicat
wastcwatcr
aclilaled\tuosc

F I CURE4. 31 Combinations of F/M andlemtion bash dissolved oxlgen conccnxrtions at $hi.h bulkingaDdnonbrlkn,s
s l u d g edsp p e ai rn c o m p l clty m i x e d,contntuousl
fedy aefurn'bl
n si ns.....................................
............. . . . . . .108
......
f I CURE4. 32 'len'poralvdnarionof F/M andSVI ar lention brsh dissolledorlgcD concentations ol {).t 100.5 m8/L

rlcuRt 4.33 Effectofdissolredorlgen uprdrcratcon SVI ofa pulp ald paperwlslcsarcfactlvatedsludgelrealDcDr

F I CURE4. 34 R c l .ti o n s i rlbpe tw e eSS

r Vl i .a n da er i onbl si nmi xhg cha..cteri sti .s.
.................................... . . . . . .I. .t .0. . . . . . . . . . . .
F I G URE4. 35
f I G URE4. 36 SolublcCOD uptaked{rntg batchsubsrrate uprde expcrimcnts virh CSTRandrrcbic tctcctorlctivared

f I CURE4. 37 Rcspirlrionutcs duing batchsrbstmteuplakeerperihcntswith CSTRlnd aenbir selcclorlctivared

f I CURE4. 38 M a x n n (na c c tl tcu p ra k ra

c tc so l Z. r.!ri g(ru \l 2l N i n barchtests.......
.. ....... .........
.....,.
....... . . . . . .I l3
HCURT4. 39 Prcdicled relatn)iship ol stady stltc grcwthmtc ^td'
atrdl )pe
acetate conoenlration ibr Type021NxndZ ru,ri3.rd,..,..,,.114
FtcuR[4.40 Dual culture.hemostaL sro*th ofZ. fo,ris.rd rnd Type02lN with iilemitrcnr fccdingofcarbon

F I CURE4. 4I Sv l a sa fu n c l h ro f rn o x l . s e l e c rdei l l uci L$l bl c C OD concentrari or.... ...........................

.. ........ . .. . . . . . . .I t 5
F I CURE4. 42 S VI a sa fu n c l i o n o f a n o x i cs e l e c l or
el l hcnt ctdtco! gl ucose concerl nti (r1..........
..,...,,..,..,...
.. ....... . . . . . . . I t 5
F I CURE4. 4: J Ag o b i cs c l c c to f-i c ri v asre
h d Ses ubstrall pl e xke$d oti l i /uri on
nechani sms......................,..,. . , . . . .t.16 ..........
FtcuRt4.44 A n o x i cs c l c c to ra c ti v astel udd g es u bstrate uptuke
andurl l i Tl ti on
mcchani snh........................... . . . . . . .t.16 ..........
FICURE 4.45 Anacrrbicsclccro.lctivatcdslLrdge $bstrareuprakernd utilizatid ncchanisnsinwl!in8 pollpho\drarc

fIGURE
4.46 Anaerobi.sclecl(rsrbsltutclpilkc rnd L,tilization
nrechanisnrinvolvjnSrltcogcn terrnentarior
lnd

FICURE
4.47 R e l n ti o n s hoi fpS Vl :1 l orc l a ti l cs i zcof i nhi 0l rcfi ti onbasi co' npMtrcnt................................ . . . . . . . . . . . 1t ll
FICURE
4.48 Etlbctof inhialcontuc(zonel-iM on SVI lor 0cturcdiiirid contncrroncs .. . .. .. .. ... .. . I t9
FICURE
4.49 M o d i l i c a ti oonf i fu e r a rd l tA S i n trdducti ul poi nrs k, ri f ri oi rbA si ns,
H l nri l roi O11.............
. ...... . . . . . . l2l
.
FICURE
4.50 Mo n th l y v c rn g cS Vl .H n rn i l l (nOH r. . l l )82l hnnrgh19119..............-....-...-.-... ...... . . . . . . I. 2l
.
FtcuRt
4.sr Efitd of irllucnt fow on 12
FICURE
4.52
FtcuRt4.s3 Q)riiSrfurionof oroxic sclcctorsyslern thc lli-Cir) | drl. ClrckrmN Cbunry,OR.......... .. .. .. .. .. .. .. ......123
FtcuRt4.54 Pcrii)mMccof lnoxic schctofrI rltcTri,Cir) pl:rnr,Cl(knmns Counly,OR........... . .. . .. .. ... t23
FtcuR[4,s5 Q rrl i g u h ti (l ro fl n l c rc b i cs c l c e toH (i vai ed
sl rdgcs]sLenr
ul nryci tc!i l l c,
A R ..... ........
........ .. ... . .. .. . . . . l?4
TIGURE
4.56 Eilicr oi tlnuftNr)bicsclcchronrhcSVIof r|lcHypr.io0orygcnircrivltcdslid8cpltuI nr Lo! An!clc\.( A..... t25
F I G URE4. 57 Modillcrti ol derdrir)rhasirto !n No{ic sclc.iorcoiliguratinrir rhc 2lklAvcnuc

F I CURE4. 58 Eltcc(of lelecor \ysrernon SVI, Ml,SS.RAS SS.rnd rir uscar tu 2-ld Avcnuc\r

F I CURE4. 59 Variriionof $pcnra(anrsu\pended vith mcu vclocir! gr lienLt(ir l.r (liltu\cd dil
s)lid conccnxation

FtcuRt4.60 E fttc lo f D rc acnc l lrc s i d e n crietn eoi di \per\ed!$p€ndcdsol i dsi r adi varcd
sl udge.
....................... . . . . . . . . 130
FtcuRt4.61 t j i l a c l o fMg S O re l d i ti o n o ' r2 h ou.acri vrredsl udgcserrl cdvol umcduri ngncl dri ah.......... . . . . . . . . . . . . . . . . . . . . . . . . . . t 3

F I CURE5. 1 Ph a s ceo n tft\tmi c ro e ra p oh f\ n o c r i di ol brm

rnd M. //nn?11dftxur\...................................... . . . . . . ll_r
,.,....,,.,,
f I G URE5. 2
F I CURE5. 3
F I CURE5. 4 Eflectof lilhing on lotal \olidsconr.iningIeepalC 620 rr Vidor vallc! RAS.................... ........ .. ............. l16
F t c uRt 5. 5 E ffe c o t ffo a n i n go n k rtl l \o l i d sc o ntl i ni ng
l gcp.l C 620arE B MU DR A S ................................. .. .. .............l-
...16
HCUR[ 5. 6 D e l a i l so f a e ra ri ob. a s i no u rl e ts
a rd \ccondl rycki ti cr ol el noN s...........................................
.. .. ........... .l.. .ll. . . .
FtcuR[5.7 Etcct of fotunlrappingon nocardidinmo,!!nism popul.tionsin bench{caleacLilaLed dudsc unii\.......... .. . 1.18
F I CURE5. 8 F,ftbclof foad mpping oullet!o. surfue &cunrulatiorof lbam on pasesofrn aefuLidb ir..... .. ..1.18
F I G URE5. 9 Mclhodsof dlsposalof foamremoledfron lreathenlb!si,rs.Mcthodsrcprcsenred bt shaded drer\ crn

F I G URE5. 10 C h e n ro s tac ot n l i g u ra ti oannsdd e i r effechon G d,,a,zzgrcw 1h,,..,...................................... . . . . . . . . . . . . . . . . . . . . . . . . . . . i

f I CURE5. 11 Rehliveamountsof lbm produced on fte surfrceofa 2,loordeepfoan tcst columnard a 15 tuor deep
a e rd ti ob. a s i nw i l h th cs a m cn o c a rdi ofbnnorgani snc.rccnl rJLi u\ ...............
......................... . . . . . .1.
. . .10
.....
f I CURE5. 12 Predicted rclationship of groivlhratcsto acetare co..enbalionfor (;. adr4. srains,Typc021N.and
FrcuRt5.13 Compdison of nocardiofom organhm populatioN at a rarS€ of MCRTSlnd tem?enturesjn bench
sludgeunitsof the city of SanFtucisco andcountyof Sacmento in Califomia.. .
scaleactjvated . l42
HCURT5.14 Schematicdiagramof scumnow! and recyclestreus (a) Southeastplant. San Francisco.CA and
... .... .
(b) rcgional{aslewatertrcatnenrplantin Sacrmenio.CA................................-.......... . . 1,42
FICURE5.I5 Relationshipof nocardioibm organismmdimum net growth rate in activatedsludgeto temperatDre - 143
5.15 Eftcct of pH on nocddioforn organisn populationsin bench scale-activatedsludgeunits . . . . - 144
'IGURE5.17 Conpanson of C. aro.@ and Z. /dnifeld tansient (ate acelateupt ke lates ove. a rangeof diludo.
fIGURE

FtcuRt5.18 Efect of aercbic selecroron nocddioforn organjsmpopulationa! a 5-day MCRT . . .. .. . . .. .. . 145

FrcuRt5.19 tstrecrof acrcbic selecloror nocddiotom orgmism populationat a l0-duy MCRT .. .. . .. .. .. . . 146
-
ftcuRE5.20 l f o o x i c s e l e c roorn n o c d di ol omorgani sm
E fi e c o popul ati ons...... ............ . - 146
FICURE5.21 Transientstateacelateuplakeby G. arozz ASF3underaorobjc.anoric,andanaerobic conditions.... - 147
IIGURE5.22 Effect of lcmperaiureon lhe wshodt MCRT for enhancedbiologicdl phosphateremovaLin activated
. l u d g e.. .. .. .. ............
.......
fIGURE5.23 L i m i ri n g M C R T v a l u e s fo rn i l ri l i cal i on,E B P R .andnocatdi ofom8rew thoverarangcoften per ar ur es- . . 149
FICURE5.24 Rcsull*of full scaleanac.obicselectorexperiments conducted at SanFrancisco in 1994and 1995 . . . . .. .I50
FICURE5.25 Re*ullsofaU full scaleanaerobic selector ei(perimenh at SanFrancisco from 1987through1995 ...... 15
FICURE5.26 MCRT and tempcraturercgiors for EBPR, nilrillcalion and nocardiofom lrowth in anaerobicselcctor

5.27 variation of alcrage nocardioformcountsand elnuent solubleonhoP concentrationswith MCRT fof full
FICURE
at S anFranchco.
s c n l ea n a c ro bsi ce l e c l oerx p e ri mcnts C A . .. . . .. ..... ....... . . . . 152
flcuREs.28 Planvicw of aerationbasinswith seleciiv.foamwasting,UtoyCreek,GA.... . . . . .. . . . .... . 153
flcuREs.29 Nocardiotarmfoarn contrcl delice for surfacechlorination !i the 23rd Avenuewastewaierrcdment Planl in

5.30 Effcctof cationicpolymcrAdditionon nocardiotbmconcentadonandaemtionb$in foaD coverage,

FICURE
T e m i n a l h l a n dPl a n t,L o sA n g e l es. .. .
C A . ..................... .... ... ..... 155
5.31 Effecton oerationbssinfoamcoverage
FIGURE ofdosing I mg/L cationicpolymo at activaleddudgeplanrin

5.32 Conpnrisonof manualconkol (1966)andautomaticcontfol(1997)of MCRT at the SanJoseisanta

FIGURE Clara
...... . . . . . . 156
on the scltlingofactivaledsludgcdominated
ftcuRl s.33 Effecrof unelatedzonesondacrutionba$inconfigufatjon

FICURE 5,34 . .......

A e n ti o n b a s i n a n d s e l e c to .s y stemal l heU OS A ,V A p14n1.......... ...... .. . . 158
-
HCURE 5,35 Bilccrof selccloroperation on SVI at theUOSA,VA pran . .. . . . . .. .. . 159
FICURE 5,36 N o n h s i dp€l a n ta e ra ti obns s i nw i thaerobisel
c cctor OK pl ant ....
at tbcTul sa, . ..... ..... 16{)
FlcuRt 5.37 Nocardioformfoam otlcct$ on percenttotal solids and ternperatureproliles in an anacrobicdigesi,er.

HCURE asa irrction ofmixing l'tchnique.. . . .

ral. in exp€rimentdan4robicdigesters
5.38 Foan accumulation .... . 16l
Listof Tables
T A B LT1. I CauscsandEffectsol Microbially'Related AcrivaledSludgeSolidsSeparatior
Prob1ens........................................2
T A B LE1. 2 C o n rp a c ti o i a n d S e ti l i n g l n te r ferenceC ansedbyFi l amentousOrgani sns.................... . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

T A B T E2. 1 T o ta E f,i l a n e nLt e n g rh(T E FL)Measurement

l i te n d e d Method............................................. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
TABLE2,2
TABLE2.3 N o c a rd h fo mO rS a n i s Fn i l a me nCounnng
l Techni 9ue.... ....................... . . . . . . . . . . . . . . . 1l
TAELE2.4 M i c tu s .o p iO
c b s e rv a ti T
o ro
n u b l e shootiGui
ng de.....
....... . .. ....... .........
....... .. . . . . . .14
.
TAELE2.5
TABLT2.6
TABLE2.7
TABLE2.8
TABLT2.9
T A E t t 2. 10
T A E LE2. I 1
T A B LE2. I 2
TAEtE2.r3
TABLE2.14
TABLE2,15
TABLE2,16 F l o c C h l ra c tc l i z d ti o i /F i l u m c nn)usOfgani sml denti l l .ul i onw (trkshcci ........................ . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
TAEIE2.'t7 FlocChlruclcrizatjor/filnm.nt(rs Organismldenlillcalionworksheet(Page2)......... ....... ............... . .. .. .. . .26
TABTE2.I8 SummAryor'TypicalMorphologicnl a d Slnini g Chancteristics
oi I.ilamenbusOrgnnirnsCom m l!

TABLE2.I9 s rl l u s o fA d i v a r€ d Sl rr| g e F i l a m ertousOrgani smTnxoromy....................................... . . . . . . . . . . . . . . . . . . . . . . . . . . . .

TABLT2.20 P rc d o mi n aHnitg h eLr i l e f_ o n nO s bscfl cdrtV afi ous
A oti vntcd
S l LrdCOrgnni
c Londi
c ng
Lcvcl s.......... . . . . . . . . . . . 5l
TABLE2.2I
TABLE2,22 S ti rrc d S[rd C e Vo l u m c Iftl c x ri S S C onccD t0ri oD of3.5A /1.(S svl ,{)............................ . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
TABLI2,23
TABLE2,24 Fo0mir8Tcstib. HiShSolidsSlmplcs(Mixcd l-jquorar Anac()bicDi.scsloQnlcrlt ........ ........... . .. .. ....54
TABLE2.25 Alk!-ScltzcftrFoamingTcsriix Inllucnland EfnucntStlca ......... ..-.-.. . ..-.-.. .54
TABLE2.26
IABLE2.27
TABTE2.28

TABLT3,1 l-ilamentAbuidAnceii U.S.BulkineandFoamineAciiv.tedSlLdecPl.nts .. .. ................ ................ . .. .. .. . an)

TABtt 3,2 Comparisoiof DomntanrFihmenlousOr8.nismsin BDlkirSandFoanrine Adivxi.d Sludecircm
S e !e ra l c o u n rrl e s ................ . . . . . . . 61
TAELE
3.3 Occurrcncc.ndRaDkofFilanrcntTypcsCausingBulkirrgin Pu\r and Pal)crWaslewaler ActilatedSlldge

TABLT3.4 and Rankoi Filrnent Type!Csusingtsullnrgii PulpandPaperwaslewarer

Occuncnce AclivaredSludge

TAELI3.5 of r,ilaftentous
Compari$r of Causes Bulkingid PDlpaodPaperWaltewaler
ActivrredSludgeSystems,

TABTE
3.6 rielationship
oi MCRT andFAI ro SpecincFilamenlous Orguismsin Activ.tedSlL'dec.... 12
.................................
TABTE
3.7 Condnions ii Which.1.tr.rzrr W.s theDomiranlFil.mcntousOrganlsmln L.boratoryScalcAclivated

TAEIE3.8 Associltcdwilh Filament()us

Surm.ry of Conditions organismCrowthin Acti!atcdSlt|dge..........................
....75

TAEIE4.1
TAELE4.2 AnnualUsagcandcost of Rulling ControlChenicalsdLLheA.tivaledsludgePlanrof sroh BrewincCo..

TAELE
4.3 U s c o fH y d ro e e n Pe ro x i d e fo rFi l anenrousB ul ki ngC onrrol ........................................ . . . . . . . . . . . . . . . . . . . . . . . . . . . .
TABTE
4.4
TASLE4.5 Compeison of Decay Ratesfor Actil"ted Sludgefton CompletelyMir@dard Ano c Sel@torSystem

TABt[ 4.6 Efiectivene$of Sele€tonin Contro]lin8Filamentous

Organisfrs ..........................................................................
I 18
IABLE 4.7 D€sign,OperatingPa.ameten,and Resulh of SelectorAciiv.led SludSeP.ocessesat th€ t-eopoldsdorfSuSd

rA8[E 4.8 Denitrification R es fof l]p. 021N, C. lruz. Strairs. Z rdiS?a, od an Aciivated SludgeDominated

TABLE4.9 DSs. FSs.andESSConcentrations b€foft andaft.r Clarin€rModiicationsat Greeley, CO ...............................

129
TABLE4.10 ClarifierT$t Result!ftom the CentralMadn Sanitadon Asencvin Califomia...............................
......................129

TAELE5.1 St€adyStateDaiafor G. arnara,Strains,

Typc021N,andZ. rumi8eru.................................................................
140
TAEIE 5.2 Wash-outMCRI (MCRTUsedlbr Nocardioform OrganisnControl)arActivaredSludgePlanr!....................... 143
TABIE 5.3 A€dvatedSludgePlant,with SurfoceWasringin Opcratjo ........................................
153
 Listof Abbreviations
a OxygentnnsfercocfllcicnlBNR mL Milliliter
APE Alkyl phenolelhoxylate pl, Micrclrter
BNR Biologicalnutrientrcmovai ML Mired iiquof
BoD5 5-daybiochemical oxygcndcmand MLSS Mixedliquor susp€nded solids
'C Degrees Celsius MLVSS Mixed liquor volatilesuspended solids
cBoD Carbonaceous BOD Millimeler
Centimeter mM Millimolar
CoD Chemicaloxygendemrnd MMF Ma{imum monthfiow
CSTR Completely stired tanft reacbr pM Micromotar
DAF Dissolvedair flotatiolr irg Microgram
DO Dissolvedoxygen pm Micron(micrometer)
DSS Dispersed suspended solids Ir,,,". Maximumgrowthrate
DSVI Diluledsludgevolumeindex N Nitrogen
EBPR Enhanced biologicalphosphorus rcmoval Na ometer
ESS Eflluentsuspended rclids NPE Nonyl phenolethoxylak
FC Fil:rmentcounr ORP Oxidadonreduclionpotenlial
FA4 Foodio-microorganism rAlb ortho-P OrlhophosphaF
(FA4)r Inirialcontacizonef,7M P Phosphorus, phosphale
FISH Fluorcscent l,1ri/& hybfidization PAO Polyphosphate accumulatiog organism
FSS Fk)cculated suspended solids PHA Polyalkanoic acid
fl F(nl, fccl PHB Poly-p-hydroxybutyric acid
F-IU li)rmrriDc turbidiryunits PVC R)lyvinylchloride
G Rootmcansquarevelocitygrudi€nt R Rccyclcratio
g Gn'm RAS RclunrActivdlcdsludgc
Spd Gallonspcr day RBC Rotrtingbiologicalconlactor
h Hour rpm Rcvoluliorspcr miuole
hp Hors€powcr s Sccond
HRT Hydraulicrcsidcncctime SBR Scqucncing batchrcaclor
i.d. Irsidc dinmctcr sp..spp. Spccic.\l-ycic\
lcz lnitial contrcrzonc SRT Solidsrcsidcncctimc
ln. lnch SS Suspcndcd solids
kg Kilogram SSVI Spccificsludgcvolumcindex
K" Half saturation cocllicienl SV,o Sclllcdvolumcal 30 min
K,,' Half saturation cocfncicnrlbr DO SVI Conlcntionalsludgcvolumeindex
Kr. Half saturution coclllcicnifbr aceiate SSVIT5 Sludgcvolunc indcxat 3.5 g SS/L
kW Kilowatt Svl" MallorySVI
L Liter TEFL Totalextended filamcntlcnglh
- lb Pound TF/SC Trioklinefihcr/solidscon|acl
m Meier TKN TotalKjeldahlnitrogcn

Cubicmeter TSS Totalsuspendedsolids

M Molar UV Ultra vrolet
MCRT Meancell residence dne VSS Volatilesuspendedsolids
mg Milligram WAS Wasteactiyatedsludee
MG Million gallons w/v Concentationin lveightper volume
MGD Miltion gallonsper day Y Yield
rnin Minute
Tableof Contents

IV SolidsSeparation
Problems
in Termsof FlocStructure
............................ ....-.........-.....3

V DifferentialionofMicrobialandProcess-RelatedSoli&SeparationProb1emi.......................................

Chspf€r2
ll.

t2

,.'''',''',,'',,35
IIL Rapid,Nonspecific
BulkingControlMethods,. ....................78
a. UpperOccoquanliewageAudroity(LJOSA).vA................................................................
{ter'1ierrncntPhn1.111sr,
b. No hsideWastet! OK.......................................
............................159
 So l i d sS e p a ra ti o nP ro b l e ms

Y()u can obserle d loL bl $xtching prcscntsa summaryoflhe causesand eftectslbr the Inost
conrmonlyidentified micfobially relatedsolids separation
Yogi tlawrence Peterl Ber.a, 1925-

I. INTRODUCTION I II, ACTIVATED

SLUDCEFLOC
Theactivated sludgeprolessalwNysconsists ofiwo liquid
Mosr aclivalcd sluclgcs)lids scprrltion t()blems can be
streamunit pro.esses a biologicrlconversn)n ofpollut (o thc naturcofthe acti,,aledsludgc fio.. A typic,rl
relalcd
antsjn a biologicalreactu andsolidsseparatntr. Lrsully
aclivrtcd sludgc has r widc rangeofprrticle sizes from
in a gravityclarifier.The functionsof biokrgicllconvcr-
singlc brclcr;n with dime sions in the afproximrle range
ijon and solidslepnratirnrcln hc combincdin r sinslc (8ocs) that can rcach
of0.5 k) 5 $ln upto largeaggregates
unit in batchopernted prolesses
strchasscqrcncingb,rtch
sizesgrente.than I lnm (l0il0 Um).
reactor!(SBR9.The secondrryclariliercanbc rcPlnccd
by a flotalion separ,rto(linrited use) or .r mcmbr{nc Activated shrdgeltocs are made up of biologicil ard
oonbiological components. The biological conrponent
A prinrnrycl4rifreris oftcn in\'olvcdin thc lk)w corsists ofa wide v.riel) ol bncleria.lungi. protozoaand
rcheme.The aerationhasinpro!idcsAncnvironmcn( Ior lome metazDa.The nonbiologicalcomponcn(is m. c up
the removalandnansfofrnarion ol hothsolublcandprr- oi inorginic and organicp.rrticu[rlcs.Thc h.Nisoi lhc l]oc
ticularepolluirntshy a nlixcd.mdvariAblcconso(iumo{' tuppcarsto bc hctcrotfophic brctcrir. Gcncft such rs
micro-nndmacro{rgrnismIcrllcd .tcti!.tlcd$ludgc.Thc Pft udoturklt, Achn kibader, l laroha(t?riLur. Akili-
secondruy clari6eftur s€ttlingphAsc in.rnSBR)p()!idcs X? ?t, Atrhnbattu4 Chtono@s, ,ir(l 7trn\k).d \Di^s atld
a quiescenienvironment thrr rllows thc rctivlrlcdsludgc Bhri, 1964rl'ike, 1972;Tabor,1976)h.r!e beensuglested
solidsrc floccul0teand then scpffnrcliun lhc trcatcd usi[8 chssicol iriolatio[ and colture methods.
wasiewlterby grn!ity sedinr€nhtion. Thcobicclivcol lhis Frr l ) .uf E el ri on(th" l c (rngl ( d,N -thrnl ngcr ganr r m .
b0sinis to prcvide! clarified(low susFndcdsdid (SS), Z'()t!lou tutrigtftt. wis lhc xolc basil ol lhc irclivatcd
low urfbidity)overdowcrlled th€ secondrryclllucntrnd sludgc Uoc htvc bccn discountcd.although/oogk)crs orc
athickened(high SS)underflowcalledlhcrcturnrctivdcd obscrvcd olrcn ir activntcd sludgc (Willi,'rrs and Unr.
sludge(RAS),Fof efficienttreunnenl.holh thc {cr tion lg83) ard rcprcscrl thc \olc sign that lhc so-callcd"rclcc-
b$in andihe secordaryclariier musttunctionsali\laclo- tff oficct" exists (van Nickcrk er nl.. 1987) or lh.rt thc
rily. Factors
afiectingbothbnnogicaloxidalionandsolids organi cIoadi ngi r hi gh. R eoentw ork uti l i ri nS nr oder n
separadon are inrporiantir detcminingovcr,rllproccss molecular biological tools such as gene probes (wagncr
etliciency. er al ., 1994rB ooder nl ., 19951S nai dr
ei al .. 1997)is vr sr ly
The purposeof this nrlrnunlis to dcscribcactivatcd expandnrgrnd ch.nging lhis listbecauseil is now known
.olid' .enrrJIi,'n
tf'hl(nF. Ji.ru* rh<ir(ru**. thai classical microbiological isolition orethodslail to
'ludge
presenta rationalapproachfrn prohlemdiagnosis.rnd deiecra grert majoriryofrnicroorgrnisnrsin environrnental
discussmeihodsfor preventing thesefroblemsby propcr sanrplcsand bixs lhc typc\ ofmicRnrginisms cnumerated.
designandoperatiororfor solvingthcnrwhcnLhcyoccur. In thc abscnccol chcmicrl rddition. rctilatcd sludge
The rs rllunrredhhctull)wll (J\e hi{ r'i... SS devclopedon municip.rlwastcwntcrcont.rinfr(nn 60 to
'nrnuJl
90% volatile matter Besides micfoorg.rnisns, activated
sludgeflocs containorganioand inoBanio pafticles.6bers
II. SOLIDSSEPARATIONPROBLEMS
lionl the irconing wastewater.and exocellular "poly-
Thereareseveraltypesof aciivatedsludgesolidssep.ra mers that play a role in biol'tocculation (Unz and Faffah.
lion problems.TheseproblenNare typioally namedin 1976r Tago and Aiba. 1977i Brown and L€sler, 1980t
rermsof theellbcis rhattheycausein thetrcatmentprocess N ol ak rnd H rugan.1981;B runsetal .,l 992;U rb aineral. ,
andbecaxse of lhis, thet definiiionsarenot alwaysprc 1993i Jorand ct a1.. 1995i Higgins .tnd Novak. 1997c1.
cise.Thephysicatandnicrobiologicalcauses for eachof These bn)polymers.ue conrposedmostly of petcins .rnd
theseproblemshavebeenfairly \ ell defined.Tablel.l
 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
P r oblem s

TABTE1.1
ActivatedSludgeSolidsSeparationProblems
Causesand Effectsof Microbially-Related
Problem Cause of Problem Effect of Problem

Dispersed growth Microorganisms aredispersed,forming only small clumps Turbid effluent; no zone settling of activated sludge
or single cells

Slime,viscous or zoogleal Microorganisms uuepresentin large amountsof exocellular Reduced settling and compaction rates; virtually no solids
bulking; nonfilamentous material; in severecases,exocellular material imparts a separationin severecases,resulting in overflow of sludge
bulking slimy orjelly-like consistencyto activated sludge blanket from secondaryclarifier; viscous foam may appear
in poor sludge dewatering

Pin floc Small, compact, weak, roughly spherical flocs are formed; Low sludge volume index (SVI) and turbid, often high SS
larger flocs settle rapidly; smaller flocs settle slowly effluent

Filamentous bulking Large amounts of filamentous microorganisms present High SVI with very clear supernatant;low RAS and WAS
bridge between the flocs or create diffuse flocs, solids concentrations; in severecases,the sludge blanket
interfering with compaction, settling, and thickening overflows the secondary clarifier; solids handling
processesbecome hydraulically overloaded

Blanket rising Denitrification in the secondary clarifier releasespoorly A scum of activated sludge forms on surface of the
soluble N2 gas which attachesto activated sludge flocs secondary clarifier and on aeration basin anoxic zones
and floats them to the secondary clarifier surface

Foam/scum formation
Caused by (i) undegraded surfactants and by
(ii) nocardioforms, M. pamicella, or type 1863
Foams (scums) can float large amounts of SS to surfaces
of treatment units; nocardioforms andM. panicellafoams
are persistent and difficult to break mechanically; foams
accumulate and can putrefy; secondaryeffluent SS can be
elevated; foams can overflow tank freeboards

The basis of activatedsludge floc formation lies in the

abilities of microorganisms to stick to each other and to
nonbiological particles.Microbial adhesionmechanisms
have been studied widely but still are not fully understood.
It appears that exocellular biopolymers form bridges
between microorganisms; these biopolymers typically
contribute 15 to 2O7oby weight of the MLSS. (Urbain et
al., 1993). At the approximately neutral pH values typical
of activated sludge, these polymers carry net negative
charges. It is thought that divalent cations such as Ca2+
,, and Mg2+ interact with the negatively charged polymers
Filament
to form bridges that allow the cells to adhereto each other
(Figure 1.1).
When built up by biopolymer bridging of relatively
spherical microorganisms, the flocs themselves will be
roughly spherical in shape (Figure l.2a). To form the
@C Polysaccharide
irregularly-shaped flocs often seen in activated sludge,
Y Protein other ingredients - filamentous organisms - are
/
!! Divalent metal
required. They provide networks or "backbones" for the
flocs. The networks direct floc growth into shapesother
FIGURE 1.1 Proposed roles of polysaccharidesand proteins than spherical (Figure 1.2b) and allow the flocs to grow
(biopolymers) and divalent cations in bioflocculation. Filamen- larger (Sezgin, l9'/1; Sezgin et al., 1978). Becausefloc
tous organisms are shown also. They bond to the floc-forming strengthdependson the integrity of the biopolymer bridg-
organisms with biopolymers and provide floc "backbones." ing, it is possible for strong and weak flocs to exist both
(From Higgins, M.J. and Novak, J.T. (1977c), Characterization
with and without filamentous organisms in them.
ofexocellular protein and its role in bioflocculation, J. Environ.
Eng., 723, 4979. With permission.)
Parker (1911) showed how filamentous organisms
could preservethe integrity of activatedsludge flocs under
S olidsS epar ati o n
Pro b l e m s

FIGU RE 1 .2 Phasecontrastmicroscopicappearances of strong activatedsludgeflocs: (a) sphericalflocs containingno filamentous

organisms;(b) inegularly shapedfloc containing filamentousorganisms.(Original magnification 100x.)

FICURE 1.3 ElTectof shearrate on 1400-pm activatedsludgeflocs: (a) G = 10.5 sr; (b) G = 79 s-r. (From Parker,D.S. (1970),
Characteristics
of Flocs in TurbulentRegimes,Ph.D. dissertation,Universityof California,Berkeley.With permission.)

conditions of increasingshear.Figure L3 shows that as polymer bridging and filamentousorganismnetworkideas

the mean velocity gradient (related to degree of turbu-
lence) is increased,the floc-forming bacteria can be
strippedaway from their filamentousorganismbackbones.
Supportfor the effect of filamentousorganismson the
structureand shapeof activatedsludgeflocs was provided
by Lau et al. (1984a)who grew a floc-forming bacterium
isolated from activatedsludge (Citromonas sp.) in dual
axenic culture with a filamentousstrain of Sphaerotilus
natans, also isolatedfrom activatedsludge.When either
organism was grown singly in chemostat culture, the
aggregatesformed were compact and roughly spherical
(Figure 1.4a).When grown togetherin approximatelybal-
ancedgrowth,irregularlyshapedflocsreminiscentof those
observedin activatedsludgewere formed (Figure l.4b). A
filamentous microorganismnetwork can be seen inside
theseirregularly-shapedfl ocs.
discussedabove.
A. DrsprnsroGnowrH
Dispersedgrowth is causedby the absenceor disruption
of exopolymer bridging so that microorganismsdo not
stick to each other (Figure 1.5). There are many (and
poorly understood)causesfor dispersedgrowth, including
selectionof nonflocculatingbacteria(single cells and/or
filaments) at high growth rates (Bisogni and Lawrence,
1971;Parker,1983;Choa and Keinath,1979),high con-
centrationsof monovalentcations (e.g., K* and Na*) rel-
ative to the concentrationsof divalent cations (e.9., Ca2*
and Mg2*) (Higgins and Novak, 1997b1'Murthy and
Novak, 1998; Novak et al., 1998; Murthy et al., 1998);
and deflocculation by poorly biodegradable surfactants
and toxic materials (Bott and Love,2002).

IV. SOLIDSSEPARATION
PROBLEMS B. Vrscous BulrrNc
IN TERMSOF FLOCSTRUCTURE
Viscous bulking (Hale and Garver, 1983) or zoogloeal
The activatedsludge solids separationproblems described bulking (Eikelboom and van Buijsen, l98l; Eikelboom,
in Thble L I can be interpreted in terms of the exocellular 2000) is causedbv excessiveamountsof exocellularmaterial

M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
FIGURE 1.4 Phasecontrastmicrographsof effects of filamentousorganismson floc structure:(a) aggregatesin a pure culture of
an activated sludge floc former; (b) aggregatesin a dual culture of a single floc former and a single filamentous organism. (Original
magnification 100x.) (From Water EnvironmentFederation.With permission.)
FIGURE1.5 Phasecontrastmicroscopic of dis-
appearance
persedgrowth.(Originalmagnification
100x.)
that may or may not be associatedwith zoogloealgrowths.
Dispersedand flocculent microbial cells are surrounded
by large amountsof water-retentive,exocellularbiopoly-
mers. Since this condition may producea viscous,poorly
settlingand poorly compactingactivatedsludge,it is pos-
sible that it constitutesone of the conditions previously
referredto as nonfilamentousbulking (Pipes, 1979).
This condition can be visualized microscopicallyby
reversestainingwith India ink (SeeTable2.9).Figure 1.6a
shows an unstainedwet mount of an extremely viscous
activatedsludgeviewed at a magnificationof 100x using
phasecontrastillumination. Notice that the flocs seemto
contain massesof dispersed cells that have smooth
rounded margins.
When a drop of India ink is addedto a wet mount of a
normal activatedsludge floc, the carbon black particles in
the ink penetratedeeplyinto the floc, largely obscuringmost
of its internal structure(Figure l.6b). When large amounts
ofexocellular materialarepresent,the carbonblack particles
do not peneffatethe floc (Figure 1.6c and Figure 1.6d). In
Figure 1.6d, individual rod-shapedbacteria surroundedby
large amountsof exocellularmaterialcan be seen.The pres-
Pr oblem s
ence of excessiveamount of exocellular material can be
confirmedby analyzingthe activatedsludgefor polysaccha-
ride and protein(SeeThble2.12 andThble2. I 3).
Activated sludge treating domestic wastewatertypi-
cally containsapproximately 15 to 2O% carbohydratein
its VSS. Activatedsludgessufferingfrom viscousbulking
can have carbohydratelevels of up to 90Voof the VSS.
Jobbagyet al. (2001) showedthat in a nutrient-deficient
activated sludge that did not contain any filamentous
organisms,the SVI increasedwith increasingexocellular
polysaccharide content.(Figure 1.7).
C. PrN Floc
When only floc-forming bacteria are present,activated
sludgeflocsareusuallysmall(up to about7-5pm), spherical
and compact(Figure l.8a and Figure l.8b). When biof-
locculation is not well developed,these flocs can be
shearedin the turbulentenvironmentof an aerationbasin
(especiallyif it is aeratedmechanicallyor with coarsebub-
ble diffusers)and by the turbulenceinducedby pumping.
by structuressuch as free-fall weirs, and tortuouspiping.
The formation is called pin floc.
As indicatedin Table l.l, this type of activatedsludge
settlesrapidly but can produce a turbid supernatant.The
larger, more compact flocs settle rapidly; the smaller
aggregatesthat may have been sheared off from larger
flocs settle slowly and createthe turbid supernatant.
Bulrcuc
D. FrmrrarNrous
Filamentousbulking is causedby an overabundanceof
filamentousorganisms.The organismsinterfere with the
settling and compaction of the activatedsludge by pro-
ducing a diffuse (or stretched-out) floc structure or by
growing in profusionbeyondthe confinesofthe flocs into
the bulk solution and bridging betweenthem. Figure 1.9a
and Figure 1.9b illustrate a bridged floc. Figure l.9c and
Fisure 1.9d show a stretched-out.diffuse floc.
SolidsSeparationProblems

FIGURE1.6 Phasecontrastmicrographsof reversestainingof activatediludge: (a) viscousflocs in an unstainedpreparation;(b)

normalflocs stainedwith India ink; (c) and (d) viscousflocs stainedwith India ink. (Originalmagnification100x,(a), (b), and (c);
1000x,(d).)(Seecolorinsertfollowingpage70.)

900 organisms observed in activated sludge is shown in

Table 1.2. Note that some filamentous organisms can
800
causeboth types of interference.When a filamentousbulk-
700 ing activated sludge settles, it produces a very clear, low
600 turbidity supernatant because the filamentous organism
network filters out the small particles (dispersedbacteria)
IJ 500 that can cause turbidity.
: 4oo
@ ooo E. Fonm/Scum
200 Filamentous organism-relatedfoaming problems in acti-
100 vated sludge are associatedlargely with the presenceof
nocardioforms and M. parvicella and, to a far lesser
degree,with type 1863. Other foams (scums)causedby
10 15 20 25 30 35 40 45 microorganisms in activated sludge arise from nutrient
deficiency and denitrifi cation.
Exocellularpolysaccharide
content,
% of biomass Nocardiofoms and M. parvicella possesspoorly wet-
table (hydrophobic) cell surfaces. When these micro-
FIGURE1.7 Etrect of exocellularpolysaccharide contenton organismsgrow in sufficient numbers in activated sludge,
the SVI of activatedsludge.(From Jobbagy,A. et al. (2002),
they render the flocs hydrophobic and amenableto attach-
WaterSci.Technol.,46:l-2,185.With permission.)
ment to air bubbles. The air bubble-floc aggregateis less
densethan water and therefore floats to the surfacewhere
The types of compaction and settling interferences it accumulatesand drains to form a thick, chocolate-brown
depend on the causativefilamentous organisms.The type colored float or scum. A similarlooking float can be pro-
of settling interference caused by various filamentous duced by denitrification in a secondary clarifier. The
 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

FIGURE 1.8 Phasecontrastmicrographsof pin flocs. (Original magnification 100x, (a); 1000x, (b).)

FIGURE 1.9 Phasecontrast micrographs of effects of fllamentous organisms on floc morphology and settleability: (a) and (b) inter-
floc bridging; (c) and (d) diffuse floc structure. (Original magnification 100x')

nitrate produced by nitrification in the aeration basin
servesas an oxygen sourcefor facultative microorganisms
in the sludge blanket at the bottom of the secondaryclar-
ifier. The nitrate is converted to nitrogen gas (Nr) which,
once it has saturatedthe solution, is releasedas small gas
bubbles. The N, gas bubbles are extremely efficient at
floating the activated sludge solids becausethey are very
small, are produced inside the flocs, and therefore firmly
attach to them. The gas-solid aggregateis lighter than
water, rises to the surface,and can createa floating sludge
problem. This can be exacerbatedwhen the denitrifying
sludge also has a significant filamentous organism content
becausethe filaments also help to trap the Nr gas bubbles'
Denitrification scum and nocardioform- or M. panicella'
derived scum can be differentiated. NocardiofotmlM. par-
vicella scum is associatedwith:
. Large, strong bubbles on the aeration basin
. Higher levels of filamentous organisms in the
scum compared to the mixed liquor
. A greasy-looking surfacefilm that forms during
a settlins test
SolidsSeparationProblems

Nutrient deficiency can be accompaniedby a sticky,

TABTE1.2 high SS scum likely causedby exocellular polymers pro-
Compactionand SettlingInterferenceCaused duced by activatedsludgebacteria under nutrient deficient
by FilamentousOrganisms conditions. This type of foam (scum) dewaters poorly
compared to the types discussedearlier.
FilamentousOrganism Bridging Open Floc Structure

Hali scomenobacter hydrossi s Yes Yes

V. DIFFERENTIATION
OF MICROBIAL
Microthrixparvicella No Yes
NostocoidalimicolaI andII No Yes AND PROCESS.RELATEDSOTIDS
Nostocoidalimicola lll Yes No SEPARATION
PROBLEMS
Sphaerotilusnatans Yes No
ThiothrixI andIl Yes No One of the most common results of the microbially-related
Type021N Yes No activated sludge solids separation problems outlined in
Type0041 Yes Yes Table 1.1 is that secondaryeffluent SS concentrations
I}pe 0092 No Yes increase.Dispersedgrowth, deflocculation of flocs by sur-
Tlpe 0581 No Yes
factants or toxicants, viscous and filamentous bulking,
'Ilpe 0675 No Yes
denitrification and nocardioform or M. parvicella foaming
Type0803 Yes No
can all produce elevated secondaryeffluent SS levels. In
Type0914 Yes Yes
'Ilpe 0961 Yes No
addition to these causes,process- and equipment-related
Tlpe 1701 Yes Yes
factors also can contribute to increases in effluent TSS
Type1851 Yes Yes concentrations.Theseinclude:

Hydraulic short circulating in secondaryclari-

Denitrification scum is associatedwith:
. Small gas bubbles in the secondaryclarifier
. No difference in filamentous organism abun-
dance between scum and mixed liquor
. Separation of settled sludge into floating and
settled layers during or after a settling test.
fiers that can decreaseclarifier HRT and scour
particles of activated sludge from the settled
sludge blanket
Turbulent conditions induced by aeration
devices,weirs, drop structures,tortuous piping,
high velocities and mixed liquor pumping that
shear activated sludge flocs

Type 1863 foam is white-grey and collapseseasily. It
appearsin aerationbasinsand in secondaryeffluents when
a plant is operatedat mean cell residencetimes (MCRT)
of approximately 2 d or less. Tlrpe 1863 organisms are
presentin foam at much higher levels than in mixed liquor.
Wahlberg et al. (2001) devisedtesting proceduresthat
allow one to differentiate between microbial and process-
related causesfor increasedeffluent SS levels (see Table
2.26, Table 2.27, and Table 2.28). The use of these pro-
ceduresfor problem diagnosis is presentedin Chapter 3.
 Methods
2
You know my methods,Watson.
Sir Arthur ConanDoyle,1859-1930,TheCrookedMan
I. INTRODUCTION
This chapterpresentsthe specializedmethodsrequired for
the diagnosisand resolution of activatedsludge solids sep-
aration problems. Microscopic evaluation of activated
sludge characteristics,filamentous organism levels, and
floc and filamentous organism types are discussedin great
detail. Chemical analytical methods for carbohydrateand
protein are presented,together with a method for differen-
tiating betweenbound and dissolvedbiopolymers.Lastly,
physical techniquesfor determining settling and foaming
characteristicsof activatedsludgeand field testing meth-
ods for differentiating between the effects of microbial
and process-relatedfactors on activated sludge solids sep-
aration are discussed.For more "routine" methods, the
readeris referred to standardtexts such as StandardMeth-
odsfor the Examination of Water and Wastewater(1998).
EXAMINATIONMETHODS
II. MICROSCOPIC
This section contains two parts. The first is devoted to
filament counting methods;the seconddescribesmethods
for the microscopic examination of activated sludge for
floc and filamentousorganismcharacterization.
A. Frmiurrur
CourunncMrrnoos
This sectionpresentstwo filament counting methodsfor
filamentousorganismsthat causebulking and a method
for counting nocardiofbrms in activated sludge.
1. Total Extended Filament Length
This methodis detailedin Table2.1. As the nameimplies,
the method determines the total length of filamentous
organismsextendingbeyondthe confinesofthe flocs into
the bulk solution. The analysis is performed on a wet
mount.The methodwas developedby Sezginet al. (1978).
2. Filament Count
This is a simplified method developed by the laboratory
staff of the San Jose/SantaClara Water Pollution Control
Plant in California to estimate the quantity of filaments
extending from flocs. This method employs an eyepiece
containinga graticulewith a hairlinedrawn on it. A known
volume of sample is placed on a slide and coveredwith
a cover slip. The entire width of the cover slip is observed
in a seriesof consecutivefields and the number of times
that fllaments intersect the hairline in each fleld (the "fi1-
ament count") is observedand totaled for the entire width
of the slide. The method is detailed inTable 2.2.
3. Nocardioform Filament Organism Counting
The rapid assessmentof nocardioform organism popula-
tions in activated sludge can be made by a nocardioform
filamentcountingtechniquedevelopedby Vega-Rodriguez
(1983)and Pitt and Jenkins(1990).SeeTable2.3. Other
methods for determining nocardioform levels in activated
sludge are either laborious and insensitive(e.g., plating
on selectivemedia and counting colonies; Wheeler and
Rule, 1980) or inappropriate(e.g., foaming tests influ-
encedby physical or chemical factors,equipmentdesign
and operation,and nocardioformorganismlevels).
Pitt and Jenkins'techniqueconsistsof microscopically
countingthe numberof branchedGram-positivefilaments
of length 2l pm in a Gram-stained,diluted, mixed liquor
sample.Filament numbers are assessedby counting the
number of intersectionsthey make with three equally-
spacedlines drawn acrossthe microscopeslide holding
the Gram-stainedpreparation(Figure 2.1). Nocardioform
counts are expressedas the number of intersectionsper
gram of VSS. The method is detailedin Table 2.3.
A similar techniquewas usedby Mamais et al. (1998)
to count M. parvicella in activated sludge. The method is
identicalto that usedfor nocardioformswith the exception
of Step l0(c) in which all intersectionsof Gram-positive
nonbranchedfilaments)3 um in lensth and <1.5 um in
width are counted.
B. Floc nruo FtLrutrurous Mtcnoonclutsitr
Cnnnncrrnrznrron
1. Introduction
Microscopic examination of activated sludge is useful for
determining the physical nature of the activated sludge
floc and the types and abundanceof filamentous organ-
isms. It generally yields information related to activated
sludge behavior in solids separationprocessesbecausethe
physical propertiesof the activatedsludge revealedduring
10 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
P r oblem s

TABTE2.1
Total ExtendedFilamentLength(TEFL)MeasurementMethod
1. Transfer 2 rnL of a well mixed activated sludge sample of known TSS concentration using a wide-mouth pipette (0.8 mm diameter tip) to
I L of distilled water in a 1.5-L beaker and stir at 95 rpm on a jar test apparatus (G = 85 s-') for 1 min.
2. Using the same pipette, transfer 1.0 mL diluted sample to a microscopic counting chamber calibrated to contain 1.0 mL and cover with a
glass cover slip.
3. Using a binocular microscope at l00x magnilication with an ocular micrometer scale, count the number of filaments present in the whole
chamber, or a known portion of it, and place these in the following size classifications: 0 to l0 pm, 10 to 25 1tm,25 to 50 pm, 50 to 100 pm,
100 to 200 pm, 200 to 400 pm, 400 to 800 pm, and greater than 800 pm. Measure filaments of greater than 800 pm in length individually.
4. Express results as total micrometers of filament length (TEFL) per gram or milliliter of MLSS:

Total extended filament length (pm) in the 1.0 mL diluted sample x dilution factor
TEFL, pm/g MLSS =
MLSS concentration, g/L

TEFL, prm/ml- MLSS = Total extended filament length (pm) in the 1.0 mL diluted sample x dilution factor

Source:From Sezgin,M. et al. (1978), J. Water Pollut. Control Fed., 50,362. With permission.

TABLE2.2
SimplifiedFilamentCountingTechnique
L Transfer 50 pL mixed liquor sample to a glass microscope slide.
2. Cover completely with a 22 x 30 mm glass cover slip.
3. Using l00x total magnification and starting at the edge of the cover slip, observe consecutive fields across the entire 30-mm length of the
cover slip.
4. Using an eyepiece ruled with a single hairline, count the number of times that any filamentous organism intersectswith the hairline for each
field.
5. Sum the number of intersections for the entire length of the cover slip. This is the filament count (FC). To express the FC as unit filament
count, perform the following calculation:

FC x numberof fields in width of slide

Unit filament count, number/prl -

Source: From Beebe, R.D. et al. (1982), paper presented at 53rd Annual Conference of Water Pollution Control Federation, St. Louis, MO. With
permission.)

microscopic examination determine its settling and com-
paction characteristics.
Only a limited amount of information can be obtained
on biological or biochemical activity from a microscopic
examination. It is difficult, if not impossible, to determine
whether an activatedsludge is healthy or unhealthy,active
or inactive, young or old. For rational diagnosis and rec-
tification of activated sludge solids separationproblems,
microscopic examination and filamentous organism char-
acterization are necessaryand invaluable.
This section presents an outline of the microscopic
examination procedure, the type of information that can
be obtained from each phase of this examination, the^
details of techniques used, and the characteization of
filamentous organism types'
2, sampling points
Take mixed liquor samplesat points of good mixing either
from the effluent end of an aerationbasin or from the mixed
liquor channel between the aerationbasin and the second-
ary clarifier. Take mixed liquor samples from below the
surface to exclude any foam or other floating material. If
foam or scum is present,take a separatesampleof the foam
or sum from one of the following points: (l) the surface
of the effluent end of the aerationbasin, (2) the surfaceof
the mixed liquor channel,or (3) the surfaceof the second-
ary clarifier. Exclude subsurfaceliquid from foam or scum
samples.Although foam and scum can be thick, viscous,
and diffrcult to transfer to sample bottles, do not dilute
them becauseone observationthat may be important is the
relative abundanceof filamentous organisms in the foam
or scum compared to the underlying mixed liquor.
Some activated sludge process modifications such as
s*p feed and contact stabilization involve more than one
aeration basin. In such systems,the activatedsludge in all
basinsusually has similar floc and filamentousorganism
characteristicsso it is not necessaryto sample all of the
basins - the effluent end of the main aeration basin will
suffice. Some systems have aeration basins in which the
conditions are different. For example, for plants with
Methods 11

TABTE2.3
NocardioformOrganismFilamentCountingTechnique"
1. Blend 400 mL of mixed liquor (or diluted mixed liquor) of known VSS concentration for 2 to 3 min at low power in a blender.
2. Prepare 5 to 10 scrupulously cleaned and degreasedfrosted microscope slides by marking the edges with a glass scribe at three equally
spaced points along their lengths.
3. On each slide place 80 pL of blended mixed liquor using a micropipette and spreadit evenly over the entire nonfrosted area of the slide.
4. Air dry the slides on a perfectly level surface.
5. Microscopically examine the slides at 100x magnification using phase contrast to check for even solids distribution over the slide.
Discard slides showing uneven distributions such as clumping, bare spots, or accumulation of solids along the edges.
6. If fewer than five slides remain, repeat Steps I through 5 to obtain five satisfactory slides.
7. Gram stain using the Hiicker modification (Table 2.6).
8. Count five slides at 1000x using oil immersion and direct illumination.
9. Use a microscope eyepiece graticule ruled with a hairline.
10. (a) Locate a scribe mark on the edge of the slide.
(b) Line up the eyepiece hairline with the scribe mark.
(c) Count any intersection with the eyepiece hairline of Gram-positive branched filaments > 1pm in length.
(d) Move across the slide to the opposite edge, counting a1l intersections of Gram-positive filaments >l pm in length with the hairline.
(e) Repeat steps 10(a) through 10(d) at the two other scribe marks on the slide.
(f) Average the number obtained for these three counts and express the results as number of intersections/g VSS.
(g) Repeat steps 10(a) through 10(f) for four more slides.
11. Averagethe resultsof 10(0 and 10(g).
12. The calculationis:

Averagenumber of intersectionsx 10" (pL/L)

Nocardiform organism count (number of intersections/g VS S) =
80 pL MLVSS(s/L)

" See Figure 2. I .

Souce: From Pitt, PA. and Jenkins,D. (1990), Res.J. Water Pollut. Control Fed., 62, 143. With permission

sequences ofanaerobicand aerobiczones,somefloc char-
acteristics(e.g.,the presenceof Neisser-positivestaining
cell clumps) may be different in the two zones.
Where an activated sludge plant consists of separate
parallel systems,floc characteristicsand filamentousorgan-
ism abundancesand types may differ betweenthe systems,
especially if the return activatedsludge (RAS) streamsdo
not mix completely. In this situation, take samples from
each system.For two-stageactivatedsludge systems(e.9.,
a carbonaceousfirst-stagefollowed by a nitrification sec-
ond-stage),take a separatesample from the aerationbasin
of each stage.Some activatedsludge systemstreat waste-
water that has been pretreatedin anothertype of biological
treatmentsystem,e.9., a lagoon,biofilter, or anaerobicpro-
cess.The pretreatmentunit may seedthe activatedsludge
with filamentous organisms. To determine the extent of
seedingtake a sample of the wastewaterentering the acti-
vatedsludgesystemand a mixed liquor sample.Also sample
any side streamsenteringthe aerationbasin (e.g.,anaerobic
digestersupematant,aerobicdigesterdecant,centrate,etc.)
to determinewhether they are sourcesof filaments or other
materials.
3. Sampling Frequency
Sampling and microscopic examination frequency will be
dictated by the circumstances.Use daily examination for
on-siteanalysisduring critical periods(e.g.,when bulking
sludgeoccurs; during RAS chlorination for bulking con-
trol; and during periods of experimentaloperation).Use
a frequency of about once every MCRT for routine char-
acterization. Use sampling frequencies of weekly to
monthly or seasonallyfor off-site examination(e.g.,by a
contractlaboratory).Match the sampling frequencywith
the severityof problemsencountered, the needto establish
an operating history, and the budget available.
4. Sample Transport and Storage
Examine samplesas soon as possible.When examination
is on-site,a delay of severalhours is inconsequential;for
more lengthy intervals, store samples at 4"C. Transport
samplesfor off-site analysisin small plastic (no glass!)
containers. Do not fill the containers more than half full
to prevent samplesfrom becoming septic.A 5-mL dispos-
able plastic transfer pipette is a suitable and economical
container. Fill the pipette bulb less than half full and heat
sealits tip. Do not freezeor chemically preservethe sample
becausethese procedurescan alter both floc and filamen-
tous organismcharacteristics.
Sample examination and data interpretation become
increasingly diffrcult as the time between sampling and
examination increases.Samplesfrom low organic loading
(F/IvI) (high MCRT) plants maintain their characteristics
longer than samples from plants with high F/M (low
MCRT). Analyze high MCRT sampleswithin 7 to 10 days
of sampling;analyzelow MCRT sampleswithin 3 to 4 days.
12 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
Mixed liquor
#
<..-f.-::a-
rF tt'l
Microscope /
FIGURE 2.1 Schematicoutline of nocardioformorganismcounting method.(From Pitt, P.A. and Jenkins,D. (1990), Res.J. Water
Pollut Control Fed.,62, 143. With permission.)
With the availability of expressmail and overnight delivery
seryices,thesetransporttimes should be easily achievable.
Avoid the following cornmon mistakes when sending
samples for analysis:
1. Container is not properly sealed.Air transpor-
tation involves pressurechangesthat encourage
sample leakage. It is very difficult to analyzea
half-dry sample that must be scrapedfrom the
inside of an envelope (delivery services do not
appreciateleaking samples either!).
2. Sample container is completely full. Besides
encouraging leakage caused by pressure
changes,septicity that influencessome filamen-
tous organismand floc characteristicscan occur.
3. Sample size is too large. When our laboratory
receives a completely-filled 5-gal container
dripping with septic activated sludge, we won-
der whether the sender wants an activated
sludge microscopic analysis or thinks he dis-
covered a new (though not very cost effective)
method of sludge disposal.
4. Samplesare sent in ice chestswith ice or cool-
ing agents.This is an unnecessaryexpense.
( "tmeasurement
Sample
Y
oitute (ifneeded) and blend
\ /
- tf
I
0
Spread80 pL evenly
on microscopeslide
{
Gramstain
1(
^'
Number of intersections= 4
Filamentous microorganism staining reactions can
change during prolonged sample storage.If long periods
are anticipated between sampling and examination, pre-
pare two air-dried smearson microscope slides at the time
of sampling and send them with the samples. This will
preserve the original Gram and Neisser staining charac-
teristics. Mark the slides with sample identification, date,
and a G or N (to indicate Gram or Neisser stain) on the
same side of the slide where the smear was made. Follow
the same procedure when samples cannot be analyzed
immediately upon receipt.
5. Microscope
Use a research-grade,phasecontrast microscope with lOx
and 100x (oil immersion) phase contrast objectives that
yield magnifications of approximately 100x and 1000x,
respectively.Phasecontrast objectives yielding 200x and
400x magnification are useful but not essential.Use phase
contrast becausebiological materials have very low con-
trast when viewed with direct illumination. Phasecontrast
illumination reveals much more detail in such low contrast
materials.
 M et hods 13

use a color slide fllm such as Kodak KR-64 for

daylight-balancedlight sources and Kodak
KPA for tungsten-balanced light sources
. Digital camera from which images can be
printed out or read directly into an electronic
file

Use the trouble-shootingguide (Table2.4)and list of "do's

and don'ts" (Table 2.5) to minimize and diagnoseprob-
lems with microscopes.

7. Staining Procedures

The Gram stain (Table 2.6) and Neisserstain (Table 2.7)

are usedroutinely in floc and filamentousorganismchar-
acterization.Additional procedureswith more specialized
uses include a test for sulfide oxidation to intracellular
sulfur granules (S test, Table 2.8); an India ink reverse
stain (Table 2.9) to detect large amounts of exocellular
polymeric materials (slime) in the floc and on {ilament
t_ surfaces;a stainingprocedureto detectintracellularstor-
age products such as poly-p-hydroxyalkanoates(PHAs,
Table 2.10); and a sheathstain using an 0.17oaqueous
Crystal Violet solution (Table 2.1l). These tables also
includea descriptionof the Anthronetestfor determining
the carbohydratecontentof activatedsludge(Table2. I 2)
FIGURE2.2 Moderntrinocularphasecontrastmicroscope
and the Lowry methodfor determiningits proteincontent
cquipped with digitalcamera. (Courtesy of OlympusAmerica
(Tabl e2. l 3).
Inc.Withpermission.)
Methodsfor measuringthe free (detachable) and cell-
A mechanical stage is essential fbr controlled scan- bound exocellular polymer in the activatedsludge arealso
ning.Usea microscope lvith a binocular head (or trinocular included (Table 2.14). These testscan be used to provide
headif photography is desired)and a built-in light source. additional confirmation of the presence and the natureof
Insertan ocularscale into one of the eyepieces and cali- large amounts of exocellular polymer observed micro-
brate it at each magnifrcationwith a stage micrometer. scopi cal l yby l ndi a i nk reversestai ni ng.
Microscopes with the features described above cunently
costabout$5000USD or more.An exampleof an excellent B. Sample Preparation
microscopefbr activatedsludgeexaminationis the Olym-
pus Model BX 4l (Figure2.2). Upon samplereceiptor within severalhours befbreexam-
Since many observationsrequiredapproachthe reso- ination, spreadI drop (approximately0.05 mL) of well
lution limits of a light microscopeand becausemany mixed sample evenly over approximately50% of the area
featuresare difficult to detect,maintainthe microscopein of each of two 25-mm x 75-mm microscopeslides.Use
excellentcondition.Regularlyadjustthephaseringsusing slides with frosted ends.Do not heatfix the slides.Allow
a centcringtclescope;keep the objectivesclean; and pro- them to air-dry at room temperature.Mark the slideswith
vide a dust-freeenvironment.Use professionalservicing a sample identification code and a G or N (to indicate
about every 5 years. Gram or Neisser) on the side on which the sample was
placed.Performthe Gram and Neisserstainingprocedures
6, Cameras outlined in Table 2.6 andTable 2.1.
Withdraw one drop (approximately0.05 mL) of well
A microscopewith a trinocr.rlar headis requiredfor micro- mixed samplewith a clean,disposablePasteurpipetteand
photography.Cameratypesinclude (in orderof increasing place on a 25-mm x 75-mm microscope slide. Place a
cost, satisfaction,and convenience): 22-mmx22-mm (No. l) glasscover slip on the drop and
pressdown gently on the cover slip with a blunt object.
. Polaroid instant film camera Remove the liquid expelled from the sides of the cover
. 35-mm camerawith a built-in light meter and slip with a tissue.Alternatively,placethe preparedsample
replaceable,fine ground-glassfocusing screenl on a paper towel. Fold the paper towel over the slide. Turn
14 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABTE2.4
MicroscopicObservationTroubleshootingGuide
Problem Possible Solutions

Image is blurred at l00x magnification. Focus condenser.

Image of phase rings is visible in field of view at 100x.
Microscope can focus at 100x but not at 1000x.
Microscope can focus with 10x objective. When 100x objective is
rotated in, it touches the slide immediately or touches the slide before
the user can focus.
User cannot seemuch detail at l00x magnification; slide looks "washed
out."
Phase contrast image shows dark background and bright objects.
Colors are distorted on stainedpreparations(assumingslides are stained
properly).
User cannot see anything.
Center condenser.
Check phase rings with centering telescope.
Oil on 10x objective: unscrew objective and look through it in bright
light. If oily, clean with liquid lens cleaner and lens tissue.
Oil on condenserlens: remove and clean with liquid lens cleaner and
lens tissue.
Oil on objective or condenserlens. Clean as detailed above.
Slide is upside down (cover slip on bottom). Make a new slide. Clean
oil off stage and condenser lens.
Cover slip is too thick. Use a No.l thickness cover slip.
More than one cover slip on slide.
Phase condenseron direct illumination. Change to correct phase
contfast setting.
Phase condenser is on wrong setting. Correct setting.
Phase condenseris on phase contrast setting. Change to direct
illumination.
Tum on light and/or increase light intensity.
Bulb is bumed out. Check bulb housing. Replace bulb. Hold it with a
paper towel. Never touch a bulb with your fingers (if you do, you will
see fi ngerprints forever).

TABTE2.5
MicroscopicObservationDo's and Don'ts
Use a minimum of immersion oil with l00x objective only.
Clean objectives only with lens tissue.
Do not get oil on the 10x objective, condenser,or stage.
Do not pull slide off the stage before you wind down the stage and/or rotate out the objective
Do not rub your eyes before washing hands.
Prepare a thin wet mount by pressing down gently on wet mount with a paper towel.
Remember that microscopic examination can be highly subjective.
Do not make up your mind about what you will see before you look at a sample.
Do not solicit prior information about what someone else thinks is in the sample.

the slide and towel over once so that the cover slip now
faces downward, with a paper towel layer over and under
it. Pressdown on the paper towel layer covering the back
of the slide. This expels excess liquid and blot dries the
expelled liquid in a single operation, as well as producing
a thin preparation.Avoid using thick preparationsbecause
long filamentsat high power can passin and out of focus
along their length. In addition to the annoyance,the focus
change is very hard on the eyes.
When processing down a slide, physical resistance,
the inability to produce a thin preparation, uneven expul-
sion of liquid around the perimeter of the cover slip, or
cracking of the cover slip is an indication that the sample
contains fairly large solid (nonbiological) particles, for
example, activated carbon, sand grains, resin beads, and
pieces of ceramic.
9. Floc Characteristics and Overall
Filament Abundance
Examine the wet mount under phasecontrast illumination
at 100x (using the lOx objective) magnification for the
following characteristics:
 Methods 15

TABTE2.6
Gram Stain,Modified Hiicker Method
Reagents Prepare fresh every 3 to 6 months.
Solution I Prepare Solutions A and B separately,then combine them.
Solution A = 2 E Crystal Violet + 20 mL ethanol 95Vo.
Solution B = 0.8 g ammonium oxalate + 80 mL distilled water.
Solution 2 | g iodine + 2 g potassiumiodide + 300 mLdistilledwater.
Decolorizingsolution Ethanol95Vo.
Solution 3 l0 mL Safranin O (2.5Vowlv in 95Voethanol) + 100 mL distilled water

Procedures:
1. Prepare thin smears on microscope slides and thoroughly air dry. Do not heat fix.
2. Stain I min with Solution 1; rinse I s with water.
3. Stain 1 min with Solution 2; rinse well with water.
4. Hold slide at an angle and decolorize with 95Vo ethanol added drop by drop (not in a
continuous stream) to the smear for exactly 25 s. Do not over-decolorize. Blot dry using
bibulous paper.
5. Stain with Solution 3 for I min; rinse well with water; blot dry using bibulous paper.
6. Examine under oil immersion at 1000x magnification with direct illumination (not phase
contrast). Blue-violet is positive; red is negative (Figure 2.17).
7. Test each batch ofGram stains on cultures with known Gram staining reactions.

TABLE2.7
NeisserStain
Reagents Prepare fresh every 3 to 6 months.
Solution 1 Prepare Solutions A and B separately.
Solution A: 0.1 g Methylene Blue + 5 mL ethanol 95Vo+ 5 mL glacial acetic acid + 100 mL distilled water.
Solution B: 3.3 mL Crystal Violet(llvo wlv in95vo ethanol) + 6.7 mL ethanol9lVo + 100 mL distilled water.
Mix two parts by volume of A with one part by volume of B.
Solution 2 33.3 mL Bismark Brown ( 17o w/v aqueous) + 66;7 mL distilled water.

Procedures:
1. Prepare thin smears on microscope slides and thoroughly air dry. Do not heat fix.
2. Stain 30 s with Solution 1; rinse 1 s with water.
3. Stain I min with Solution 2; rinse well with water; blot dry.
4. Examine under oil immersion at 1000x magnification with direct illumination (not phase contrast). Blue-violet is positive
(either entire cell or intracellular granules); yellow-brown is negative (Figure 2.18).

a. Floc Size c. Protozoa and Other Macroorganisms

Measure 10 to 20 flocs and place them in the following Determine the presenceand types of protozoa and other
size categories based on their minimum dimensions or macroorganisms(e.9.,rotifers, nematodes);seeFigure 2.31
diameters if they ale approximately spherical: through Figure 2.35.

d. Nonbiological Organic and lnorganic Particles

small <150pm Confirm that a particle is nonbiological by observing it
Medium 150-500pm under direct illumination. Much of the biological material
Large )500pm will not be visible while nonbiological (especially inor-
ganic) material is readily observed (Figure 2.4).
b. Floc Characteristics e. Bacterial Colonies
Determine whether flocs are round or irregular and com- Determine the presenceof bacterial colonies that appear
pact or diffuse; determine whether texture is firm or weak to be composedof one type of microorganism.These may
(Figure 2.3). be fingered zoogloeas (Figure 2.5a and Figure 2.5b),

l\ _
16 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABTE2.8
Sulfur OxidationTest(STest)
Test A
Reagent: Sodium sulfide solution (NarS.9H2O), 1.0 g/L; prepare fresh weekly.

Procedures:
1. On a microscope slide, mix 1 drop of activated sludge sample and 1 drop sodium sulfide solution.
2. Allow to stand open to the air for 10 to 20 min.
3. Place a cover slip on the preparation and gently pressto exclude excesssolution; remove expelled solution with a tissue.
4. Observe at 1000x using phase contrast. A positive S test is the observation of highly refractive, yellow-colored
intracellular sulfur granules inside the filaments (Figure 2.22b, Figure 2.23b, and Figure 2.23d).

Note: If results are inconsistent, use the sulfur oxidation test (Test B below).

Soarce.'Modified from Eikelboom, D.H. and van Buijsen, H.J.J. (1981), Microscopic Sludge lwestigation Manual,TNO
Research Institute for Environmental Hygiene, Delft, The Netherlands. With permission.

TestB
Reagent: Sodium thiosulfate solution (NarSrO..5HrO), I g/100 mL; prepare fresh weekly.

Procedures:
1. Allow activated sludge sample to settle; transfer 20 mL of clear supematant to a 100 mL Erlenmeyer flask.
2. Add I to 2 mL of mixed activated sludge sample to the flask.
3. Add I mL of sodium thiosulfate solution to the flask (final sodium thiosulfate concentration is 2mM).
4. Shake the flask overnight at room temperature.
4. Observe at 1000x using phase contrast, as in Test A.

.loarce; Modified from Nielsen, P.H. (1985), Water Sci.Technol.,1'7:2-3, 167.With permission.

TABTE 2.9
lndialnk Reverse
Stain
Reagent:India ink (water-basedblack drawing ink; aqueoussuspensionof carbon black particles; also known as China ink).

Procedures:
l. Mix I drop of India ink and I drop of activated sludge sample on a microscope slide," depending on the ink used,
the sample volume may need to be reduced.
2. Place on the cover slip and observe at l00x using phase contrast.
3. In normal activated sludge, the India ink particles penetrate the flocs almost completely, at most leaving clear
centers (Figure l.6b). In activated sludge containing large amounts of exocellular polymeric material, large, clear
areaswith low cell density will be observed(Figure 1.6c and Figure 1.6d).

" Alternatively, apply 1 drop of India ink to the edge of a cover slip under which a thin prcparation of the sample has
been made. Place a piece of absorbent paper at the opposite edge of the cover slip. This causesthe India ink particles to
be pulled through the sample. The observer can see the India ink particles spreading across the slide.

amorphouszoogloeas(Figure 2.5c and Figure 2.5d), nilri-
fying bacteria (Figure 2.6), or organisms that accumulate
polyphosphate (Figure 3.10c). In some cases, an entire
floc may seem to consist of one morphological type of
microorganism.Thistypeoffloc,mostcommonlyencoun-
tered in activated sludge treating industrial wastewaters,
is said to have poor diversity (Figure 2.7).
bacteria are motile. This can be difficult becauseconvec-
tive liquid movement under the cover slip caused by the
heat of the microscopic lamp causesthem to drift. Do not
interpret this as motility. Judge motility as motion that is
different in speed and/or direction from a general drift.
For some dispersed microorganisms (e.9., spirochaetes
andSpirillum spp.;Figure 3.1lc and Figure 3.11d),motil-

r Ceus
Dispersed
inButk
sotution flJilH:i;Hilf #1fi}j"j::-::H*:i"T:i'J;
Determine the presenceof dispersedcells in the bulk The supernatants of samples containing significant
solution (Figure 1.5). Determine whether the dispersed amounts of dispersedcells will appear turbid.
Methods 17

TABLE2.10
(PHA) Stain
Poly-p-hydroxyalkanoate
Reagents:
Solution 1: SudanBlack B, 0.3Vowlv in 607oethanol.
Solution 2: SafraninO,0.59o w/v aqueoussolution.

Procedures:
1. Prepare thin smears on a microscope slide and thoroughly air dry.
2. Stain 10 min with Solution l; add more stain if the slide starts to dry out.
3. Rinse 1 s with water.
4. Stain l0 s with Solution 2; rinse well with water; blot dry.
5. Examine under oil immersion at 1000x magniflcation with direct illumination (not phase contrast).
Intracellular PHA granules appear as blue-black granules; cytoplasm will be pink or clear.

Source: From Manual of Methods for General Bacteriology (1981), American Society of Microbiology,
Washington, D.C. With permission.

problems) if they are scored "common" or less. This

TABLE2.11
CrystalViolet SheathStain
Reagent: Crystal Violet, 0.|Vo w/v aqueoussolution.
Procedure: Mix I drop (0.05 mL) of activated sludge sample and
I drop ofCrystal Violet solution on a microscopeslide; place on a
cover slip and examine at I 000x magnification using phasecontrast,
The cells stain deep violet; sheathsare clear to pink.
method is rapid and suitable for establishing whether a
filamentousorganism is dominant or secondaryand for
determining filament responseto remedial actions.Repro-
ducibility betweenobserversis generallyto within t one
abundancecategory.With practice, it is possibleto achieve
a consistentrelationshipbetween the subjectivescoring
of filamentabundanceand measuresof sludgesettleability
such as SVI (Chapter3).

g. Effects of Filamentous Organisms 10. FilamentousOrganism Characteristics

on Floc Structure
Classify the effect of filamentousorganismson floc struc-
tures as fbllows:
. None.
. Bridging: filaments extend from the floc surface
into the bulk solution and bridge between the
flocs (Figure 1.9aand Figure 1.9b).
. Open floc structure: floc population attachesto
and grows around the filamentous organisms
leading to large, irregularly shaped flocs with
substantialinternal voids (Figure l.9c and Fig-
ure 1.9d).
h. Filamentous Organism Abundance
Use a subjectivescoring systemto determinefilament abun-
dance.Observefilamentousorganismsfirst at 100x and then
at 1000x. On the basis of these observations,subjectively
rate overall filament abundanceon a scalefrom 0 (none) to
6 (excessive).See Table 2.15 and Figure 2.8. Determine a
sampleabundancerating for all filament types togetherand
an abundancerating for each filamentous organism type.
Consider individual filamentous organisms dominant and
most likely responsible for solids separationproblems if
they are scored "very common" or greater.
Consider individual organismssecondary(presentbut
not in sufficient abundanceto accountfor solids separation
Next, determine the types and abundancesof the filamen-
tous organisms.Use 1000x phase contrast to carefully
characterizeeach filamentous organism. Look at several
filamentsof each type and expressthe observationsas an
average.Use filamentous organism worksheetssuch as the
onespresentedin Table 2.16 andTable2.1'7to record and
summarizeindividual filamentousorganismobservations.
Simplify this task by acceptingonly a limited number of
descriptions and learning to recognize features that pro-
vide clues to specific filamentous organism types. The
characteristicsare discussedbelow.
a. Branching
Report whether branching is present or absent and, if
present,whethertrue or false.True branchingshowscontig-
uous cytoplasm between branched filaments (Figure 2.9a
and Figure 2.9b). ln activated sludge, the only filament-
forming organismsthat commonly have true branchesare
fungi and nocardioform organisms. False branching
occurs when no contiguous cytoplasm is presentbetween
filaments. In a false branch, two filaments have merely
stuck together and grown outward (Figure 2.9c). In acti-
vated sludge,false branching is only observedfor sheathed
filaments. This is most commonly observed for Sphaero-
tilus natans. False branchesare observedrarely for Halis-
comenobacterhydrossis,Type 1701 andThiothrix spp.
1B P roblem s
M anua lo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on

TABTE2.12
AnthroneTestfor Total Carbohydrate
When boiled with concentratedsulfuric acid, pentoseand hexose-basedcarbohydratesform furfurals that react with aromatic amines to form colored
products whose concentrations can be measured spectrophotometrically.

Reagents:
Sulfuric acid solution: 75Va vlv reagent grade. Slowly and carefully add 750 mL concentrated sulfuric acid to 250 mL distilled water (be sure to
add the acid to the water!). Cool to room temperature before using.
Anthrone reagent: Add 200 mg Anthrone reagent; (CAS. No. 90-44-8) to 5 mL absolute ethanol; make up to 100 mL with'l\Vo sulfuric acid.
Store at 5oC and replace monthly or sooner if brown color develops.
Standard glucose solution: Add 100 mg glucose to 100 mL distilled water and add 150 mg benzoic acid (preservative).Store at 5"C. Dilute l:10
with distilled water for daily use (1mL = 100 pg glucose).
Sodium chloride solution 0.857o w/v: Dissolve 8.5 e NaCl in 1 L distilled water.

Equipment:
Thick-walled Pyrex boiling tubes (15 cm x 2.5 cm)
Water bath for boiling tubes; must be large enough to continue boiling after chilled samples and standardshave been placed in it
Ice water bath
Spectrophotometerand cuvettes

Procedures:
1. Centrifuge an activated sludge mixed liquor sample of known MLSS concentration to separatesolids. Discard the solution (centrate).Wash
the centrifuged solids by resuspending them in 0.857o NaCl solution. Recentrifuge then resuspendto original volume with distilled water.
Omit this step if the results from washed and unwashed samples are the same.
2. Pipette a range of sample volumes from 0.1 to 1.0 mL (containing 10 to 100 pg carbohydrate as glucose) into a series of boiling tubes;
adjust all final volumes to 1.0 mL with distilled water.
3. Using the 100 pg/ml glucose standard and distilled water, prepare 3 to 4 glucose standardsin the range of l0 to 100 pg glucose, each with
a final volume of 1.0 mL. Include a 1.0 mL distilled water blank.
4. Chill all tubes in the ice water bath.
5. Add 5.0 mL chilled Anthrone reagent to each tube; mix thoroughly and keep these in the ice water bath.
6. Transfer all tubes to the boiling water bath for exactly 10 min. The boiling water bath must continue to boil throughout the transfer of the
boiling tubes to it. Most problems (with variable results) encountered in this procedure are caused by an inadequately sized boiling water
bath that cools too much during transfer of the cold boiling tubes.
7. Retum the tubes to the ice water bath.
8. Measure absorbanceof all tubes at 625 nm using the distilled water tube as a blank.
9. Prepare a standard curve for the glucose standardby plotting absorbanceversus glucose concentration. This should be a straight line. Read
the sample glucose content from this standard curve. Use only sample values between <907o transmittance and the percent transmittance
produced by the highest glucose standard that lies on the straight line portion of the standard curve. Express results as pg/ml glucose.
Convert this to mg total carbohydrate per gram dry weight activated sludge, or report as percent of dry weight as glucose.

Note; After some experience with this test, the proper dilutions of the activated sludge can be determined, allowing the number of tests to be reduced.

Source: From Manual of Methods for General Bacteriology (1981), American Society of Microbiology, Washington, D.C. With permission.

b. Motility
Report whether the organisms are motile or nonmotile. If
they are motile, describe the type of motility. Only a few
filamentous organisms in activated sludge are motile.
Beggiatoa spp., Flexibacter spp., and some blue-green
Cyanophyceae bacteia exhibit gliding motility; Thiothrix
spp. and Type 021N may display limited twitching or
swaying motions.
c. Filament Shape
Determine whether organisms are straight, smoothly
curved, bent, irregularly shapedchains of cells, coiled or
mycelial (Figure 2. 10).
d. Location
Determine whether filaments extend from the floc surface,
lie mostly within the floc, or are free in the liquid between
flocs (Figure2.ll).
e. Attached Bacteria
Report whether attachedbacteria are presentor absent.If
present, determine whether growth is substantial or inci-
dental (Figure 2.12).The presenceof attachedgrowth can
indicate that a filamentous orsanism has a sheath.
f. Sheath
Report whether a sheathis presentor absent.A true sheath
is difficult to seebecauseit is a clear structureoutside the
 Methods 19

TABTE 2.13
ModifiedLowryMethodfor Protein
Reagents:
S o l u t i o n A : D i s so lve 2 g p o ta ssiu m so d iu m ta r tr a te CoH oOuK N aandl 00gN arC O3i n500mLl MN aOH anddi l utetol Lw i thdi sti l l edw ater.
Store in a plastic bottle. Discard when turbid.
S o l u t i o n B : D i s so lve 2 g p o ta ssiu m so d iu m ta r tr a te a ndl gC uS Oo.5H rOi n90mLdi sti l l edw aterthenaddl 0mLl MN aOH .S torei nap l as ti c
bottle. Discard when turbid.
Solution C: Prepare fresh daily. Dilute I volume of Folin-Ciocalteu reagent to 15 volumes with distilled water. The solution should have an
acidity of 0.15 to 0.18 N when titrated with I N NaOH to pH 10.
Standard protein solution: Dissolve 5 mg crystalline bovine serum albumin in 100 mL distilled water. Store at 4oC. Prepare fresh monthly.

Equipment:
Spectrophotometer(650 nm) and lens curvettes
Medium-weight Pyrex test tubes (15 cm x 2.5 cm) matched for wall thickness
Water bath at 50€

Procedures:
1. Centrifuge an activated sludge mixed liquor sample of known MLSS concentration to separatethe solids. Discard the solution (centrate).
Wash the centrifuged solids by resuspendingthem in 0.857o NaCl solution. Recentrifuge and resuspendto the original volume with distilled
water. Omit this step if the results from washed and unwashed samples are the same.
2. To a test tube add I mL sample containing 15 to 100 pg protein; add 0.9 mL Solution A.
3. Place test tube in the 50oC water bath for 10 min. Cool to room temperature (21 to 25.C) and add 0.1 mL Solution B.
l.- 4. Allow to stand at 2l to 25oC for at least 10 min. Add 3 mL Solution C with sufficient force to mix the test tube contents thoroushlv.
5. Place the tubes in the 50oC water bath for 10 min. Remove and cool to room temperature.
6. Measure absorbancein 1-cm cuvettes at 650 nm.
7. Concunent to analyzing samples, analyze a set of bovine serum albumin standardsand a distilled water blank.

Note: The absorbanceat 650 nm produced by the modified Lowry method is linear, ranging from 15 to 1l0 pg protein as bovine serum albumin.

Source: From Hartree, E.F. (1972), Anal. Biochem.,48,422. With permission.

cell wall. In unstainedpreparations,sheathsare most eas-
ily seen when they are empty (contain no cells) or when
some cells in the filament are missing. In the latter case,
the outline of the sheath can be seen to continue along
both sides of the empty space (Figure 2.13a and Figure
2.13b). Several features of filamentous organisms can be
confused with sheaths.
Do not mistake a yellowish "halo" around filaments
viewed under phase contrast illumination for a sheath.It
is an artifact of phasecontrastillumination. The difference
between a halo and a true sheath can be seen clearly in
Figure 2.13c. Notice that the bright halo is wider than the
sheath.Do not assumeshort, empty spacesin a filament
or at the filament apex indicate the presenceof a sheath.
The cell walls of some filamentous organisms (e.g., Tlpe
02lN) may remain after cell lysis (Figure 2.13d). Distin-
guish these from sheaths by looking for remains of
cross-walls. Note that the empty cells usually have the
same diameters as normal cells.
Use the presence of substantial attached bacterial
growth to indicate a sheath.Visualize sheathsby mixing
equal volumes of activated sludge and a 1:1000 dilution
of household bleach (57o sodium hypochlorite solution)
and allowing the mixture stand for several hours. This
treatment lyses the cells inside the sheaths,leaving empty
sheathsthat can be seenunder phasecontrast illumination
at 1000x.
g. Cross-Walls (Cell Septa)
Report presenceor absence.This feature can be variable
for certain filamentous organisms and detection depends
on the quality and adjustment of the microscope.
h. Filament Width
Using an ocular micrometer, measureboth averagewidth
and its range (pm). Determine whether the averagewidth
is >l pm (thick filament) or <1 pm (thin filament).
i. Filament Length
Report the averagelength and range of lengths in pm.
j. Cell Shape
Determine whether cell shapeis square,rectangular,oval,
barrel, discoid (resembling a stack of hockey pucks),
sausage-shaped, or irregular (Figure 2.14). Note whether
indentations are present at the cell septa (Figure 2.14c
through Figure 2.141 or whether the filament walls are
straightat the cell junctions (Figure 2.14a andFigure2.14b).
k. Cell Size
Report the averagelengths and widths of the cells in pm.
20 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE2.14
ExocellularBiopolymerExtractionMethods
Dissolved Biopolymer
Equipment:
High speed centrifuge (to 10,000 rpm).
High speed centrifuge tubes.

Procedures:
l. Pour 40 mL well mixed sample into a high speed centrifuge tube.
2. Centrifuge at 10,000 rpm for 15 min.
3. Decant supematant containing the soluble biopolymer fraction into a test tube. The supematant can
be analyzed for carbohydrate(Table 2. l2) and protein (Table 2.13), omitting Step I in each procedure.

Bound Biopolymer
Equipment:
Waring blender, 100 mL capacity.
High speed centrifuge (to 10,000 rpm).
Jar test apparatusor paddle stirrer.

Procedures:
l. Transfer pellet obtained from Step 3 of the dissolved biopolymer extraction to a Waring blender
using 40 mL l0r M NaOH.
2. Mix 3 s on low speed to resuspendpellet.
3. Transfer to a jar test apparatusand mix 15 min at 90 rpm.
4. Transfer to a high speed centrifuge tube; centrifuge at 10,000 rpm for 15 min.
5. Decant the supernatantcontaining the bound biopolymer fraction. The supematant can be analyzed
for carbohydrate (Table 2.12) and protein (Table 2.14), omitting Step 1 in each procedure.

Source: From Higgins, M.J. and Novak, J.T. (1997a), Water Environ. Res., 69,225. With permission.

FIGURE 2.3 Phasecontrast micrographs of floc characteristics:(a) compact, (b) diffuse, (c) firm, (d) weak. (Original magnifications
100x, (a) and (b); 1000x, (c) and (d).)
Methods 21

FIGURE2.4 Micrographsof nonbiologicalparticlesin floc: (a) phasecontrast,(b) directillumination.(Originalmagnification100x.)

(Seecolor insertfollowing page70.)

FICURE2.5 Phasecontrastmicrographs of zoogloeas:(a) and(b) fingered,(c) and(d) amorphous

(globular).(Originalmagnifications
200x, (a); 1000x,(b); 100x,(c); and 1000x(d).) (Seecolor insertfollowing page70.)

L SulfurDeposits m. Other Granules

Determinepresenceor absencein situ and presenceor
absenceafterperformingtheS test(Thble2.8).Underphase
contrastobservation,sulfur granulesappearas bright,
refractive,yellow-coloredcell inclusions,either spherical
(Thiothrix spp.,Beggiatoaspp.,and Tlpe 021N; Figure
2.15athroughFigure2.15c),or square-rectangular (Type
0914;Figure2.L5d).Whensulfurgranulesarevery small,
they can appearblack ratherthanyellow underphasecon-
trastillumination.Type0914doesnot respondto the S test.
Report whether other granules are present or,absett.
Commonly observedgranules are polyphosphate(Neisser
positive granules; Figure 3.10c) and PHAs (confirmed by
PHA staining;Table 2.10 and Figure 2.16).
n. Staining Reactions
Evaluateeach filamentous organism for Gram and Neisser
staining reactions by obsewing stained, air dried smears
at 1000x using direct illumination (not phase contrast).
22 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FIGURE2.6 Phasecontrastmicrographsof nitrifying
bacteria.(Originalmagnification100x,(a); 1000x,(b) and
(c).)
FIGURE2.7 Phasecontrastmicrographof floc with poordiver-
sity. (Originalmagnification1000x.)
Carefully note the general locations and lengths of filamen-
tous organismsin the wet mount and the presenceor absence
of attachedgrowth so that the filament types observedin the
wet mount can be matched to those examined in the stained
smears.Take care in making this observation becausesome
filamentous organisms can change size upon drying and
staining. For example, Type 0092 appearsmuch wider when
Neisser stainedthan in wet mounts.Ignore the Gram stain-
ing reactionsinside large denseflocs becausethey do not
decolorize properly (Figure 2.17a).
Score the Gram reaction as strongly positive, weakly
positive, variable, or negative.Most filamentousorganisms
in activated sludge are Gram-negative (Figure 2.17b).
N.limicola I and II and Tlpes 0041,0675, and 0914 most
often are Gram-positive but can be Gram-variable
(Figure2.l7c and Figure2.l7d) or Gram-negative.Tlpe
1851 stainsweakly Gram-positiveand generallyappearsas
a chain of Gram-positive"beads" (Figure 2.17d). Thiothrix
I and II, Beggiatoa spp., and Tlpes 021N and 0914 gener-
ally stain Gram-negative,but may stain Gram-positive when
containing substantial intracellular sulfur deposits.
M. parvicella and nocardioform organisms are generally
strongly Gram-positive (Figure 2.17e and Figtxe 2.17f).
Score the Neisser stain as negative, positive (entire
trichome is stained), or negative with Neisser-positive
granul es (Fi gure 2.18). Type 0092 (l i ght pur ple;
Figure 2.18d) and N. limicola I, II, and III (dark purple;
Figure 2.18e) usually stain entirely Neisser-positive.
M. parvicella and nocardioform organisms usually stain
Neisser-negative but generally contain Neisser-positive
intracellular granules (Figure 2.18b and Figure 2.18c).
Most of the other filaments may contain Neisser-positive
intracellular granules at times. In addition, H. hydrossis and
Tlpes 0675 and 0041 may have Neisser-positivefilament
"coverings" (Figure2.18f) which constitute a false reac-
tion causedby chemical precipitation of the Neisser stain
on the outside of the filament. This is observed only in
industrial wastewateractivated sludge systems.
o. AdditionalObservations
Thiothrix spp. and Type 021N may display rosettes and
gonidia. A rosette develops when filaments radiate outward
Methods 23

TABTE2.15
SubjectiveScoringof FilamentAbundanceu,b
NumericalValue Abundance
0 None
I Few
2 Some
3 Common
/ Very common
5 Abundant
6 Excessive
Explanation
No filaments observed
Filaments present, but only observed in an occasional floc
Filaments commonly observed, but not present in all flocs
Filaments observed in all flocs, but at low density (1 to 5 filaments per floc)
Filaments observed in all flocs at medium density (5 to 20 per floc)
Filaments observed in all flocs at high density (>20 per floc)
Filaments observed in all flocs (more filaments than floc and/or filaments
growing in high abundancein bulk solution)

" This 0 to 6 scale representsa 100- to 1000-fold range of total extended filament length.
bSee Figure 2.8 for examples of filament abundancescores.

from a common origin (Figure 2.19a and Figure 2.19b).
Gonidia are oval- or rod-shapedcells present at the fila-
ment tip that are distinctively different in appearancefrom
the rest of the vegetative cells further down the filament
(Figure 2.19c and Figure 2.19d). Rosettesand gonidia
indicate that the organisms are growing rapidly.
11. FilamentousOrganism ldentification
a. Using the Dichotomous Key
Characterize the filamentous organisms to genus or to a
numbered "type" using the dichotomous key shown in
Figure 2.20. This procedure is simplified in two ways.
First, only a limited numberof characteristicsare usedto
describethe organisms(Figure 2.20 andTable 2.18). Sec-
ond, only the 23 filamentous bacteria most commonly
observed in activated sludge are listed. The infrequently
observedTypes 1702 and 1852 are omitted.
To further simplify the key, severalfilamentous organ-
isms having readily identifiable specific characteristicsare
not included and are describedseparately.These include
fung| Cyanophyceae,Flexibacter spp.,Bacillus spp., and
actinomycetes.
This dichotomouskey is a modification of the fila-
mentous organism identification key of Eikelboom and
van Buijsen(1981).Changeshavebeenmadeto de-empha-
sizethe needfor the observationof cell septa(cross-walls),
which can depend on the quality and adjustment of the
microscope used, and to include some filamentous organ-
isms in the key twice where an important characteristicis
variable, e.g., the Gram stain reaction for N. limicola II
and the observation of intracellular sulfur sranules for
Types0914 and 021N.
Using this key is not without risk becausesome fila-
mentous organism characteristicsvary and the key cannot
always addressall variables. Carefully check all the fila-
ment types found by using the key (Figure 2.20) against
the typical organism characteristicslisted in Table 2.18
and presentedin the descriptionsand photographsof each
organism. If the characteristicscited in Table 2. 18 or
shown in the photographs do not correspond to the fila-
ment type determined by using the key, carefully reexam-
ine the characteristicsused in the key. For example, Type
0041 usually producesa weak Gram-positiveor Gram-
variable reaction and may be correctly identified from the
dichotomous key. However, if strongly Gram-positive, it
would be keyed as N. limicola IL Table 2.18 shows that
N.limicola II is coiled and free of attachedgrowth, while
Type 0041 is straightor smoothly curved and most often
has substantialattachedsrowth. The Gram stainedslide
should be re-examined.
Occasionally,a filamentousorganismobservedcannot
be placed in a type or genus designated in the key.
Describethe organismcharacteristicsand report it as "not
identified." Do not try to "force fit" an organism into
existing filament types.
b. Building Your Skills
One of the hardestparts of filamentous organism identifi-
cationis building the confidenceto make an identification.
To develop this confidence(l) characterizethe filamen-
tous organismsin a sampleand sendthe same sampleto
someoneskilled in the technique(some treatmentplants
in a particularareaestablishedround-robinfilament iden-
tification proceduresto assisteach other); and (2) take a
courseon microscopic characterizationof activatedsludge
and filamentousorganismidentiflcation.
The task is not as daunting as it might at first appear
to someoneassignedto a particulartreatmentplant. It is
doubtful that someonein sucha situationwould encounter
all the filamentousorganismtypes listed in Table 2.18. It
is common to deal with about half a dozenthat eventually
can be recognizedeasily.
12. FilamentousOrganism Descriptions
The individual fi lamentousorganismdescriptionspresented
here are based on our experiencewith characteristicsof
filamentousorqanismsfound in activatedsludsesall over
24 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

FIGURE 2.8 Phase contrast micrographs of filament abundancecategories using the subjective scoring system: (a) few, (b) some,
(c) common, (d) very common, (e) abundant, and (f) excessive.(Original magnification 100x.)

the world. Data from Eikelboom and van Buijsen (1981)
and Eikelboom (2000) on samples from northem Europe
are also incorporated.
a. Sphaerotilus natans (Figures2.9c, 2.13b,
2. 14f , an d 2 .2 1 a )
Filament Description - Straight or smoothly curved
filament that extends from the floc surface.Filaments are
generally 100 to >500 pm in length. Cells are sausage-
shaped with indentations at the cell septa and typically
1.6 x 2.5 pm in dimension. Cell shapecan be rectangular
when cells are packed tightly within the sheath.A sheath
is present; false branching is usually present. Attached
growth is generally absent, but may be present if the
filament is not growing. Not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics - Large size, false branching,
and sheath.
b. Type 1701 (Figure 2.21b)
Filament Description - Straight, smoothly curved, or
bent filament commonly found within the floc or can
extend from the floc surface.Filaments are generally 10 to
150 pm in length. Cells are sausage-shaped and typically
 Methods 25

TABTE2.16
OrganismldentificationWorksheet
FlocCharacterization/Filamentous

Sample number Sample location

Sample date | | Observation dato I I

Comments:

Observatlons of:
Protozoa
Metazoa
Wet mount observation: 1000xphasecontrastfor filament characteristics
1000xdirect illumination for stains

Filament number I 2 3 4 J

Branching

Motility

Filament shape

Filament location

Attachedg:rowth

Sheath

Crosswalls

Filament diameter,pm

Filament length, pm

Cell shape

Cell size,pm

Sulfur deposits

Other sranules

Gram stain

Neisserstain

Commonness

Rank

Identification

i--
I
 t

26 Manualon Causesand Control of ActivatedSludgeBulking,Foamingand Other SolidsSeparationProblems

TABLE2.17
OrganismldentificationWorksheet(Page2)
FlocCharacterization/Filamentous

Sample number Sample locatlon

Sample date Observadon dsb I I

FilamentAbundance:
n0
nI
n 2
t-l nn n 6
None Few Some Common Very Common Abundant Excessive

FilamentEffect on Floc Structure: n i'*:- f-l

t-l
Bridging
n
I
OpenFtoc
I Structure

Morphology of Floc:
nFirm
nWeak
n
Round
ntr
Irregular Compact
n
Diftuse

Freecells in suspension Neisserpositive cell clumps

Zoogloeas India ink test
Spirochaetes Chlorine damageto filaments
'Inorganic/organicparticles

FilamentousMicroorganism Summary:

Rank Abundance Rank Abundance

Nocardioforms M. parvicella

Typ€1701 Type0581

S. tutans Tlpe 0092

Typ€02lN Tlpe 0803

Thiothrix spp. Tlpe 1851

Type0041 T}pe 0691

H. hydrossis Tlpe 0675

N. lirnicola Other

Fungus Other

Remarks:
Methods 27

FIGURE 2.9 Phase contrast micrographs of branching of

filamentous organisms in activated sludge: (a) and (b) true
branching (fungus and nocardioform organism), (c) false
branching (Sphaerotilus natans). (Original magnification
1000x.)

0.8 pm to 1.0x 1.5pm in dimensionwith indentations at
the cell septa.A sheathis presentandattachedgrowthgen-
erally is present.Falsebranchingoccursrarely.Not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics- Small size,usuallysausage-
shapedcells;usuallyinsideflocswith flocsformedaround
filaments.
c. Haliscomenobacterhydrossis(Figure2.21c)
Filament Description - A rigidly straightor bent flla-
ment that extendsfrom the floc surface;may be found
within the floc or free in suspension.
Filamentsare 10 to
100pm in lengthand0.5 pm in diameter.Cell septaare
not present;however,empty spacesin the filamentmay
appearas cells.A sheathis presentand attachedgrowth
canbe present.Falsebranchingoccursrarely.Not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative. e. Thiothrix I (Figures2.14b, 2.15a, 2.19c,
Key Characteristics - A very thin, small filament
and 2.23a)
that canbe overlookedif not examinedat magnifications
greaterthan 100x;has"pins in a pincushion"appearance. Filament Description - A straight or smoothly curved
d. Type021N(Figures 2.11b,2.13d,2.14d,
2.14e, 2.15c, 2.1Ba,2.19b, and 2.22a)
Filament Description - A smoothlycurvedfilamentthat
extends from the floc surface. Filaments are 50 to
2500 pm long and I.6 to 2.5 pm in diameter.Indentations
are piesent at the cell septa. Cell shape is quite variable
(rectangular, oval, barrel-shaped).Individual cell dimen-
sions are typically 2.0 x |.5-2.0 pm. Filaments often taper
from a thicker basalregion with an inconspicuousholdfast
cell to a thinner apical region, often terminating in loosely
attached gonidia (sausage-shaped)cells. Rosettes (many
filaments radiating outward from a common floc) may
occur. A sheath is absent and attached growth does not
occur. No branching and not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Positive but variable. Intracel-
lular spherically shapedsulfur granules may be presentln
situ or after performing the S test.
Key Characteristics - One of the largestand longest
filaments. Indentationsat cell septa.Cell shapevariesfrom
rectangular to oval or barrel-shaped.Rosettesand gonidia
may occur. If growing on sulfide, intracellular sulfur gran-
ules are usually present and the S test will be positive.
filament that extends from the floc surface.Filaments are
100 to 500 pm in length and 1.6 to 2.5 pm in diameter.
The cell shapeis rectangularand cells are I.6 to 2.5 pm x
2.0 to 4.0 pm in dimension. There are no indentations at
the cell septa.Filaments often taper from a thicker basal
region with an inconspicuous holdfast cell to a thinner
28 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

FIGURE2.10 Phasecontrastmicrographs of filamentshapes:(a) straight,(b) smoothlycurved,(c) bent,(d) irregularlyshaped

of cells,(e) coiled,and (f) mycelial.(Originalmagnification1000x.)

apical region, often terminating in loosely attached
gonidia (sausage-shaped)cells. Rosettes(many fllaments
radiating outward from a common floc) may occur. A
relatively thick sheath is present generally without
attachedgrowth. Attached growth only occurs if this fila-
ment is not growing. No branching and not motile.
Staining Reactions - Generally Gram-negativeand
Neisser-negative;may stain Gram-positive when growing
on sulfide.
Sulfide Oxidation - Positive but variable. Intracel-
lular spherically shaped sulfur granules may be present in
situ or after performing the S test.
Key Characteristics- Oneof thethickestfilaments,
typically 2.0 to 2.5 pm, but sometimesas largeas 4 pm
in diameter.Intracellularsulfur granulesare usually
presentand the S testis positiveif growingon sulfide.
f. ThiothrixII (Figures2.19d, 2.23c, and 2.23d)
Filament Description - A straightor smoothly curved
filament that extendsfrom the floc surface.Filamentsare
typically 50 to 200 pm in length and 0.8 to 1.4 pm in
diameter.The cell shapeis rectangularandcellsare0.8 to
1.4pm x 1.5to 3.0 pm in dimension.Thereareno inden-
tationsat thecell septa.Filamentsoftentaperfrom a thicker
 Methods 29

FIGURE2.11 Phasecontrastmicrographsshowingloca-
tionsoffilamentousorganisms:(a) freefloating(dispersed),
(b) extendingfrom floc surface,(c) within floc. (Original
magnifications1000x,(a); 1000x,(b) and (c).)

FIGURE 2.12 Phaseconffast micrographs showing attachedgrowth of bacteria on fllamentous organisms: (a) heavy, (b) incidental.
(Original magnifi cation 1000x.)

basal region with an inconspicuousholdfast cell to a thin-
ner apical region, often terminating in loosely attached
gonidia (sausage-shaped) cells. Rosettes(many filaments
radiating outward from a common floc) may occur. A rel-
atively thin (hard to observe) sheath is present, generally
without attached growth. Attached growth may occur when
the filament is not growing. No branching and not motile.
Staining Reactions - Generally Gram-negativeand
Neisser-negative;may stain Gram-positive when growing
on sulfide.
Sulfide Oxidation - Positive but variable. Intracel-
lular spherically shapedsulfur granulesmay be presentin
situ or after performing the S test.
Key Characteristics - Thiothrixll is generally about
1.0 pm in diameter and can be confusedwith severalother
filaments of the same diameter. It differs from other sim-
ilarly sized filaments in that it is straight or smoothly
curved and extends away from the floc surface. Intracel-
lular sulfur granules are usually present and the S test is
positive if growing on sulfide.
g. Type 0914 (Figures2.15d, 2.24a, and 2.24b)
Filament Description - A straight or smoothly curved
filament that extends from the floc surface, is inside the
floc or is dispersedin suspension.Filaments are typically
50 to 200 pm in length and 1.0 to 1.2 pm in diameter.

I
I
30 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FICURE2.13 Phasecontrastmicrographsshowingappearances
of sheaths:
emptycellsin Tlpe 021N.(Originalmagniflcation1000x.)
The cell shapeis squarewithout indentations at the septa
and cells are 0.8 to 1.2 pm x 1.0 pm in dimension.A
relatively thin (hard to observe)sheathis present.Attached
growth is common but may be absent,particularly when
growing dispersed.No branching and not motile.
Staining Reactions - Generally Gram-negativeand
Neisser-negative;may stain Gram-positive when growing
in the presenceof sulfide.
Sulfide Oxidation - Positive but variable. Square-
shapedintracellular sulfur granulesmay be presentin situ.
This filament does not respond to the S test.
Key Characteristics - Square-shapedirregular sul-
fur granules. Type 0914 can be confused with several
filaments of similar size if attachedgrowth is present and
sulfur granules are absent.Its sheathis diffrcult to observe
but is indicated by the presenceof attachedgrowth. Eikel-
boom (2000) noted that [pes 0914 and 0803 are often
complementary, i.e., when Type 0914 disappears, type
0803 appears,and vice versa. Based on this observation,
he suggeststhat Types 0914 and 0803 may be two forms
of the same organism.
h. Beggiatoa sp. (Figures2.15b, 2.22c, and 2.22d)
Filament Description - A straight or smoothly curved
filament that extends from the floc surfaceor is dispersed
(a)and(b)Sphaerotilus
(c)Tlpe0041,and(d)
natonE,
in suspension. Filamentsaretypically 100to >500pm in
length and 2.0 to 4.0 pm in diameter.The cell shapeis
rectangular withoutindentationsat the septa;however,the
cell shapeis maskedwhen intracellularsulfur granules
are present.The cells are2.0 to 4.0 pm x 6.0 to 8.0 pm
in dimension.Sheath,attachedgrowth,andbranchingare
absent.Beggiatoaspp.areusuallymotileandexertflexing
and gliding motions.
Staining Reactions- GenerallyGram-negative and
Neisser-negative; may stainGram-positivewhencontain-
ing manyintracellularsulfur granules.
SulfideOxidation - Generallypositive.Intracellular
sphericallyshapedsulfur granulesmay be presentin situ
or after performingthe S test.
Key Characteristics- Gliding,flexingmotility and
largesize.
i. Nostocoida limicola I (Figure2.25a)
Filament Description - An irregularlycurvedfilament
intertwinedwithin the flocs.Filamentsaretypically40 to
100pm in lengthand0.8 to 1.0pm in diameter.The cell
shapeis oval and cells are 0.8 to 1.0 pm x 0.8 pm in
dimension.Indentations occurat the cell septa.No sheath
is presentand no attachedgrowth occurs.No branching
and not motile.
 Methods 31

FICURE 2.14 Phase contrast micrographs of filamentous

organism cell shapes: (a) square, (b) rectangular, (c) oval,
(d) banel, (e) discoid, (0 sausage,and (g) inegular. (Orig-
inal magnification 1000x.)
32 M anualo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,
FIGURE 2.15 Phasecontrastmicrographsof intracellularsulfur granules:
(d) Type 0914. (Original magnification1000x.) (Seecolor insert followingpage70.)
FICURE2.16 Directilluminationmicrograph of PHA staining.
PHA granulesare dark blue/black.(Original magnification
1000x.)(Seecolorinsertfollowingpage70.)
Staining Reactions - Gram-positive and Neisser-
positive; may be Gram-negative and Neisser-negativein
activated sludge treating some industrial wastewaters.
Sulfide Oxidation - Negative.
Key Characteristics - Typically intertwined within
the flocs; readily identified by its Neisser-positivestaining
reaction and the presenceof individual cells. This filament
can be confusedwith Type 0092 becauseboth are Neisser-
positive andM. p arv iceI Ia becalse both are Gram-positive,
and Other S ol i dsS eparati on
P r oblem s
(a) Thiothrixsp.,(b) Beggiatoasp.,(c) Type02IN, and
but N. limicola I has individual cells that are not observed
for these other filaments.
j. Nostocoida limicola Il (Figures2.10e,2.11c,
2.17b, 2.18e, and 2.25b)
Filament Description - An irregularly curved filament
that extends from the floc surface and is present within
the floc. Filaments are typically 50 to 200 pm in length
and 1.4 pm in diameter. The cell shape is oval and cells
are 1.4 x 1.0 pm to 1.5 pm in dimension. Indentations
occur at the cell septa.No sheathis presentand no attached
growth occurs. No branching and not motile.
Staining Reactions - Gram-positiveor -negativeand
Neisser-positive;generally Gram-positive in municipal
wastewater activated sludge but often Gram-negative in
industrial wastewateractivated sludge (especially in pulp
and paper wastewater activated sludge); is occasionally
Neisser-negativein industrial wastewateractivatedsludge.
Sulfide Oxidation - Negative.
Key Characteristics - Neisser-positive staining
reaction and the presenceof individual cells. If Neisser-
negative, its cell structure is usually sufficient for identi-
fication. In stainedpreparations,the structuresappearlike
"runs" in nylon stockings. Branching does not occur. An
actinomycete may be confused with this filament, but the
branching of the actinomycete separatesthese filaments.
M et hods 33

FIGURE 2,17 Direct illumination micrographsof Gram stainingof lilamentousorganisms:(a) improperlydecolorizedfloc retaining
Crystal Violet, (b) Gram-negaliveNostoctsidalimicola II, (c) Gram-variableType 0041, (d) Gram-variableType 1851. (e) Gram-
positive nocardioform organism, and (f) Gram-positiveMicrothrix parvicella. (Original magnilication 1000x.) (See color insert
following page70.)

k. Nostocoida limicola lll (Figure 2.25c)
Filament Description - An irregularly curved filament
that extendsfrom the floc surface.Filamentsare typically
100 to 300 p.rmin length and 2.0 pm in diameter.Cell
shapeis oval to discoid and cells are 2.0 pm x 1.5 pm in
dimension.Indentationsoccur at the cell septa.No sheath
is presentand attachedgrowth doesnot occur.No branch-
ing and not motile.
Staining Reactions - Gram-positiveor -negativeand
Neisser-positive;is generally Gram-positive in municipal
wastewateractivated sludge and generally Gram-negative
in industrial wastewateractivated sludge (especially pulp
and paper wastewateractivatedsludge); is occasionally
Neisser-negativein industrial wastewateractivatedsludge.
Sulfide Oxidation - Negative.
Key Characteristics - Neisser-positive staining
reaction,Iargesize and individual discoid-shapedcells. If
Neisser-negative, its cell structureis usually sufficientfor
identification. Eikelboom (2000) no longer recognizes
N. limicola Il as separatefrom N. limicola IIl.
34 Ma nu al on Caus es and Cont r ol of Ac t iva t e d S l u d g e B u l k i n g , F o a m i n g , a n d O t h e r S o l i d s S e p a r a t i o n P r o b l e m s

"rtw

''' ti::::.::.
::::t?:i':r

, r*:',:
r-
I
"sy ,i

,*
I

tr

FICURE 2.18 Direct illuminationmicrographsof Neisserstainingof filamentousorganisms.(a) Neisser-negative Type 021N, (b)
with positivegranules(nocardioformorganismsandMicrothrix purviceLla,respcctively),(d) and (e) Neisser-
and (c) Neisser-negative
positive(Type 0092 andNostocoidalimit'olu II, respectively),(f) false Neisscr-positivestain covering lilament,Type 0041. (Original
magnification1000x.)(Seecolor insertfbllowing page70.)

l. Type 0411 (Figure2.26a) Sulfide Oxidation - Negative.

Filament Description - An irregularly curved filament
that extendsfrom the floc surface.Filamentsare typically
50 to 150 pm in lengthand 1.0pm in diameter.The cells
are shapedlike elongatedrods and are 0.8 to l.2pm x
2.0 to 5.0 um in dimension.Indentationsoccur at the cell
septa.No sheathis presentand attachedgrowth does not
occur. No branchingand not motile.
Staining Reactions - Gram-negativeand Neisser-
nesative.
Key Characteristics - Long sausage-shaped
cells;
irregular lilament shapes.
m. Type 0961 (Figure 2.26b)
Filament Description - A straightor smoothly-curved
filament that extendsfrom the floc surface.Filamentsare
typically 40 to 150 pm in length and 1.0 to 1.4 pm in
diameter.The cell shapeis rectangularand cells are 1.0 to
1.4 ;rm x 2.0 to 4.0 pm in dimension. No indentations
Methods
FICURE2.19 Phasecontrastmicrographsof rosettesand gonidia:(a) rosette,(b) Tlpe 021N rosette,(c)Thiothrix I gonidia,(d)
ThiothrixII gonidia.(Originalmagnifications100x,(a); 1000x,(b), (c), and (d).)
appearat the cell septa.No sheathis presentbut a slime
coveringon the filamentthat looks like an empty"cuff'
may be presentat the filamenttip. No attachedgrowth;
no branching;not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics - Long rectangularcells;
"transparent"appearance; no cell inclusions.
n. Type0092 (Figures2.18d and 2.26c)
Filament Description - A straight or bent filament
alwaysinsidethefloc. Filamentsaretypically 10to 80 pm
in lengthand0.8 to 1.0 pm in diameter.Individualcells
usuallycannotbe seenwithin the filament.No sheathis
presentandno attachedgrowthoccurs.No branchingand
not motile.
Staining Reactions - Gram-negativeand Neisser-
positive.
Sulfide Oxidation - Negative.
Key Characteristics - Location inside the floc and
Neisser-positivestainingreaction.Filamentis often over-
lookedin a microscopicexaminationof a wet mountbut
becomesrecognizable whena Neisser-stained preparation
is examined.Filamentsappearwider (1.0 to 1.2 pm) in
35
preparations
dried Gram-andNeisser-stained thanin wet
mounts.
o. Type 0581 (Figure2.26d)
Filament Description- Curvedor coiledfilamentinside
the floc. Filamentsaretypically 100to 200 pm in length
and0.5 to 0.8 pm in width.Individualcellscannotbe seen
within the filament.No sheathis presentand attached
growthdoesnot occur.No branchingand not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics- Locationwithin the floc and
Gram-negative stainingreaction.This filamentcanbecon-
fusedwith M. parvicellabut hasa Gram-negative staining
reaction;M. parvicellais Gram-positive.
p. Type0041(Figures 2.10a,2.12a,2.13c,2.14a,
2.17c, 2.18f, and 2.27a)
Filament Description - A straight filament usually
insidethe floc. Filamentsare typically 100to 500 pm in
length and 1.8 to 2.0 pm in width. Individual cells are
square-shaped and 1.8 to 2.0 pm x 2.0 to 3.0 pm in
dimension.A heavysheathis present;branchingis absent.
Large amounts of attachedgrowth are present. The
 3O Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

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 Methods
absenceof attached growth suggestsa rapid growth rate.
No branching and not motile.
Staining Reactions - Gram-variable (usually
weakly Gram-positive) and Neisser-negative,sometimes
with rows of round Neisser-positivegranules.Occasion-
ally stainslightly Neisser-positivein industrial wastewater
activatedsludge - a false reaction causedby precipitation
of Neisser stain on the surface of the filament.
Sulfide Oxidation - Negative.
Key Characteristics - Large size, Gram-variable
(weakly Gram-positive) staining reaction, attached
growth, and location within the floc.
q. Type 0675 (Figure 2.27b)
Filament Description - A straight filament usually
inside the floc. Filaments are typically 50 to 150 pm in
length and 1.0 pm in width. Individual cells havea square
shapeand are 1.0pm x L0 pm in dimension.A thin sheath
is present.Branchingis absent;heavyamountsofattached
v growth are usually present.No branchingand not motile.
,+
S t aining R e a c ti o n s - G ra m-v a ri a b l e (usual l y
weakly Gram-positive)and Neisser-negative.
Sulfide Oxidation - Negative.
Key Characteristics - Gram-variable(usually weakly
Gram-positive)staining reaction,attachedgrowth and loca-
tion inside flocs. The filament is similar to but smaller than
Type 004 I . Eikelboom (2000) no longer differentiatesTypes
0041 and 0675 and combinesthem as Type 0041/0675.
r. Type 1851 (Figures2.17d and 2.27c)
Filament Description - A straight or smoothly curved
filament usually inside the floc. Filaments are typically
50 to 200 pm in length and 0.8 pm in width. They may
intertwine to form twisted "ropes" or "bundles". Individ-
ual cells are rectangularand 0.8 pm x 1.5 to 2.0 pm in
dimension.A thin sheathis presentand extensiveattached
growth is usuallypresent.The attachedgrowth is typically
orientated perpendicularly to the cell surfacesof the fila-
ment. No branching and not motile.
Staining Reactions - Gram-variable (usually
weakly Gram-positive) and Neisser-negative.
Sulfide Oxidation - Negative.
Key Characteristics - Formation of twisted filament
"ropes" or "bundles" by intertwinedfilaments;perpendic-
ular attachedgrowth and Gram-variable staining reaction.
s. Type 0803 (Figure 2.27d)
Filament Description - A straight filament that may
extend from the floc surface and be dispersedin the bulk
solution. Filamentsare typically 50 to 150 pm in length
and 0.8 pm in width. Individual cells are squarewith no
indentationsat the septaand 0.8 x 1.0 pm in dimension.
No sheath;no attachedgrowth; no branching; not motile.
Staining Reactions - Gram-negative and Neisser-
nesative.
37
Sulfide Oxidation - Negative.
Key Characteristics - Dispersed growth. This fila-
ment resembles Type 0675 but has neither sheath nor
attached growth. It stains Gram-negative,while 0675
stainsweakly Gram-positive.Eikelboom (2000) suggests
that Types 0914 and 0803 may be the sameorganism.
t. Microthrix parvicella (Figures2.17e, 2.18c,
2.28a, 5.1e, and 5.1f)
Filament Description - An irregularly coiled filament
found inside the floc, surroundingthe floc, and dispersed.
Filamentsare typically 50 to 200 pm in lengthand 0.8 pm
in width. Individual cells cannot be seen. Sheath and
attachedgrowth are absent.No branchingand not motile.
Staining Reactions - Strongly Gram-positiveand
Neisser-negative.Neisser-positiveintracellular granules
are commonly observed.
Sulfide Oxidation - Negative.
Key Characteristics - Strongly Gram-positive
stai ni ng reacti on; N ei sser-posi ti vegranul es; coiled
growth; inability to see individual cells; "beaded" effect
causedby intracellular granules.No branching and not
motile.
u. Nocardioforms (Figures2.9b, 2.10f, 2.149,
2.17f, 2.18b, 2.28b, 5.1a, 5.1b, 5.1c and 5. 1d)
Filament Description - Inegularly shapedtrue-branch-
ing filaments occurring inside the floc and dispersedin
the bulk solution. Filamentsare typically -5to 30 pm in
lengthand 1.0 pm in width. Individualcells are present
and shape is irregular. Sheath and attachedgrowth are
absent.Not motile.
Staining Reactions - Strongly Gram-positiveand
Neisser-negative.Neisser-positiveintracellular granules
are commonlyobservec.
Sulfide Oxidation - Negative.
Key Characteristics - True branchingand strongly
Gram-positive staining reaction. The true abundancein
activatedsludge can only be assessedafter examining a
Gram-stainedpreparation.Becausenocardioformscom-
prise many genera (e.g., Nocardia, Gordona and Skerma-
nia), characteristicsdescribedcan vary considerably.One
type, Skermania pinensis (the pine-tree-like-organism or
PTLO) is distinguishableby its characteristicbranching
(see Figure 5.lc and Figure 5.1d). Many nocardioforms
can grow in a fragmented or single cell form.
v. Type 1863 (Figures2.10c,2.14c, and 2.28c)
Filament Description - An irregularly shapedfilament
dispersedin the bulk solution. Filaments are typically
10to 50 pm in length and 0.8 to 1.0 pm in width. Indi-
vidual cells are present,and the cell shapeis oval,0.8 to
1.0 pm x 1.0 to 1.5 pm in dimension.No sheath;no
attached growth. No branching and not motile.
 I

33 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

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 Methods 39

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40 Ma nu al on Caus es and Cont r ol of Ac t i v a t e d S l u d g e B u l k i n g , F o a m i n g , a n d O t h e r S o l i d s S e p a r a t i o n P r o b l e m s

FfGURE 2.21 Phasccontlastmicrographsof: (it) Sphucro-

tiItr,stttrtttrt.v, (b)'lypc I70l, (c) Huliscomuutltu<tcr lrydnts-
, r l , r .( O r i g i r r a l m a g n i l i c a t i o n l ( X X ) x . ) ( S c c c o l o r i n scr t
firllorvingpage70.)

FlCURE2.2 2 Ph as ec ont r as t m ic r o- { r aphs(of

a): Ty p c 0 2 l N , ( b ) T y p e 0 2 l N r . v i t h s u l f l r g r a n u l e s , ( . c ) B e g g , i a t o a s p . , ( d) Be g g i a to tt
(Originalrnagnilication1000x.)(Seecolol insertfbllorvingpagc70.)
sp. rvith sullirrgranLrles.
Methods 41

FIGURE2.23Phasecontrastmicrographsof (a)Thiothrixl,(b)Thiothrixlwithsulfurgranules,(c)Thiothrixll,and(d)Thiothrixll
with sulfur granules.(Originalmagnification1000x.)(Seecolor insertfollowing page70.)

FIGURE2.24 Phase contrast of: (a)Tlpe 0914,(b)Tlpe 0914 with sulfur granules.(Originalmagnification1000x.)
micrographs
(Seecolorinsertfollowingpage70.)

Staining Reactions - Gram-negativeand Neisser-
negativewith Neisser-positiveintracellulargranules.
Sulfide Oxidation - Negative.
Key Characteristics - A flexible chain of irregular-
shapedcells often containingNeisser-positive granules.
w. Type0211 (Figure2.28d)
Filament Description - An irregularly shapedfila-
Filamentsaretypically l0 to
mentthat occursdispersed.
50 pm in length and 0.4 pm in width. Individual cells are
present, and the cell shape is oval, 0.4 pm x 0.6 pm in
dimension. Sheath and attached growth are absent. No
branching and not motile.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics - A very thin flexible chain of
cells; the thinnest filament found in activated sludge.
42 Man ua l on Caus es and Cont r ol of Ac t i v a t e d S l u d g e B u l k i n g , F o a m i n g , a n d O t h e r S o l i d s S e p a r a t i o n Pr o b l e m s

FIGURE 2.25 Phasecontrastmicrographsof: (a) No,rto-

coida limiolu l, (b) No,stocoitla
limicoltt ll, (c) Nostot'oida
limicolu lll. (Originalmagnification1000x.)

a):Ty p e 0 4 l l , ( b ) T y p e 0 9 6 l , ( c ) T y p e 0 0 9 2 , a n d ( d ) T y p c 0 5 8 1 . ( O r i g i nm
FICURE 2 .26 Phas ec ont r as t m ic r ogr aphs(of a la gn i fi ca ti o n
10 00 x.1
Methods
FIGURE2.27 Phasecontrastmicrographsof:(a)T1pe0041,(b)Type0675,(c)Tlpe1851,and(d)T1pe0803.(Originalmagnificati
1000x.)
x. Flexibactersp. (Figure2.29a)
Filament Description - An elongated,rod-shaped bac-
terium ratherthan a true filament.Straightor smoothly
curvedand 1.0pm x 20 to 40 pm in dimension.It occurs
dispersed.No sheath;no attachedgrowth;no branching.
Motile with a flexing andgliding motion.
Staining Reactions - Gram-negative and Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics- Flexingandgliding motility.
y. Bacillussp. (Figure2.29b)
Filament Description- A longchainof rod-shaped cells
ratherthan a true filament.Straightor inegularly curved
and 1.0pm x 20 to 50 pm in dimension.Occursdispersed
or attachedto the floc. A sheathmay be presentbut
attachedgrowthis absent.No branchingand not motile.
Staining Reactions - Gram-positiveand Neisser-
negative.
Sulfide Oxidation - Negative.
Key Characteristics- Gram-positivestainingreac-
tion; "chain of cells" appearance.
43
z. Cyanophyceae (Figure 2.29c)
Filament Description - Large, straight filamentous
blue-green photosynthetic bacteria (often called blue-
greenalgae),2.0 to 5.0 pm wide and 100 to 500 pm long.
They occur dispersed.No sheath;no attachedgrowth; no
branching. May have slow gliding motility.
Staining Reactions - Gram-positive or -negative
and Neisser-negative.
Sulfide Oxidation - Negative.
Key Characteristics - Large size and a distinct
green color when viewed under direct illumination.
aa. Fungi (Figures2.9a and 2.29d)
Filament Description - Large, truly branchedfilaments,
5.0 to 10 pm wide and 100 to 1000 pm long, usually
inside the flocs. A sheath is absent; however, a heavy
cellulose cell wall is usually present.Attached growth is
absent. True branching occurs. Intracellular vacuoles,
organellesand granules are present and internal cytoplas-
mic streaming occurs. Not motile.
Staining Reactions - Gram-positive or -negative
and Neisser-negative.
Sulfide Oxidation - Neeative.
44 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FIGURE2.28 Phasecontrastmicrographsof: (a) Miuothrix parvicella, (b) Nocardioforms,(c) Type 1863,and (d) Tlpe 0211.
(Originalmagnification1000x.)(Seecolor insertfollowing page70.)
Key Characteristics - Very large, true branching
filaments containing vacuoles, organelles, and granules
and exhibiting cytoplasmic streaming.
C. Pnocnrss rN IDENnFYTNG
Fuurrurous Oncnrutsrtts
Table 2.19 is a summary of the status of identifying and
classifying the filamentous organisms found in activated
sludge. More information about the identities of filaments
and types of organisms in activated sludge continuously
becomes available through the application of genetic
methods. Indeed, it is not beyond the realm of possibility
that by the time the next edition of this manual is written,
rapid test kits for identifying filamentous organisms in
activated sludge will replace the microscopic methods
described this chapter.
An especially promising genetic method for detecting
filamentous (and other) microorganisms in activated
sludge is fluorescent in situhybidrzation (FISH). A gene
probe (a fragment of genetic material approximately 15 to
30 basesin length) complementary to (binds specifically
with) a unique portion of one of the RNA molecules in a
cell's ribosomes (usually the l6SrRNA molecule) is
prepared. The probe is then reacted with a fluorescent mol-
ecule that, when illuminated with ultraviolet (UV) light,
can be visualized. When the probe is added to a sample,
it will bind only to the l6SrRNA to which it is comple-
mentary.The extent and location of the probe binding can
be detectedby observing the samplethrough a microscope
with UV illumination. FISH probes have been used exten-
sively in the characterizationand identification of activated
sludge filamentous organisms.Other examplesof their use
in activated sludge are described below.
Bouchez et al. (2000) showed that bacteria used to
"bioaugment" a laboratory sequencingbatch reactor(SBR)
activated sludge system (with the objective of increasing
its denitrifying capacity) was very rapidly eatenby stalked
ciliated protozoa.
For many years,Nitrosomon&s(ammonia oxidized to
nitrite) and Nitrobacter (nitdte oxidized to nitrate) micro-
organisms were credited with nitrification in activated
sludge. Severalworkers recently called these assumptions
into question. For example, Wagner et al. (1998) found
that in an activated sludge treating an industrial waste-
water with very high ammonia content, the predominant
ammonia oxidizer was an organism resembling Nitroso-
coccus mobilis. They were unable to find significant num-
bers of Nitrobacter spp., leading them to believe that
nitrite oxidation was carried out by another organism.
FISH studies by Juretschko et al. (1998) and Mobarry
et al. (1996) demonstratedthat the nitrifiers did not grow
Methods
FIGURE2.29 Phasecontrastmicrographsof: (a) Flexibactersp., (b) Bacillus sp., (c) Oscillatoriasp. (cyanophyceae),
fungus.(Originalmagnification1000x.)(Seecolor insertfollowing page70.)
in dispersedform and were not disfibuted evenly throughout
the activated sludge floc. Rather they grew as dense col-
onies (Figure 2.30a) and the ammonia oxidizer colonies
were surrounded by the nitrite oxidizer colonies
(Figure 2.30b). These findings have significant implica-
tions, both for bioaugmentation and for the modeling of
nitrification in activated sludge.
The discrepancy between these findings and those from
previous work in which Nitrosomonas andNitrobacter spp.
were identified as the principal nitrifying organisms in
activated sludge results from the bias introduced by cul-
turing microorganisms from environmental samples.
Because many environmental microorganisms will not
grow on laboratory culture media, the only way to be
certain whether an organism is important in an environment
such as activated sludge is to demonstrate its presence by
in situ methods that do not involve culturing and isolation.
An excellent example of mistaken identity causedby
the bias introduced by culture and isolation methods was
the conclusion that organismsof the Acinetobacter genus
were largely responsible for the phenomenon of enhanced
biological phosphorusremoval (EBPR) in activatedsludge
systems with initial anaerobic zones. Recent work has
clearly shown that this is not so. Microorganisms that are
not closely related to Acinetobqcter spp. are important in
this role (Hesselmannet al., 1999; Crocetti et al., 2000).
45
and (d)
D. Pnorozol Alro Mrrlzon
1. General
Microscopic observationof protozoa and other higher life
forms in activated sludge is a common and widespread
practice. In a very general way, the types of theseorganisms
present can be related to plant performance and effluent
quality. They are useful for toxicity assessmentbut they
are of little or no value for determining the properties of
activated sludge that influence its behavior in solids sep-
aration processes.
From a morphological view, activated sludge is a rel-
atively simple microbial community consisting of free and
flocculated bacteria (and at times fllamentous bacteria),
protozoa, rotifers, nematodes, and a few other inverte-
brates. Protozoa and other higher life forms are usually
aerobic and bacteriovorous (they eat bacteria). A few anaer-
obic flagellates and a number of saprophytic flagellates
occur in activated sludge. The saprophytic flagellates use
soluble organic matter for growth. Carnivorous free ciliates
and attached ciliates (suctorians) feed on other protozoa.
Chlorophyll-bearing flagellates are incidentally observed
and are usually derived from aeration basin walls.
Protozoa and other higher life forms may constitute
approximately 5%obyweight of the activatedsludgebiomass
and are represented by about 200 species (Curds, 1973;
 46 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE2.19
Statusof ActivatedSludgeFilamentousOrganismTaxonomy
Name in Key FISH Current Name and
(Iable 2.18) Pure Culture Probe Taxonomic Position References

S. natans Yes Yes Sphaerotilus natans Richard et al. (1982); Williams and Unz (1985a, 1985b); Ziegler et al.
(1990); Corstjensand Muyzer (1993); Wagner etal. (19941
Type l70l Ye s No Sphaerotilus spp. Richardet al. (1982);Williams and Unz (1985a);Kiimpfer et al. (1995);
Howarth et al. (1998)
H. hydrossis Yes Yes Haliscomenobacter van Veen (1973); van Veen et al. (1982); ZieEler et al. (1 990); Wagner
hydrossis et al. (1994)
Type 021N Yes Yes SeveralThiothrix spp. Howarth et al. (1999); Wagner et al. (1994); Ziegler et al. (1990);
Kanagawa et al. (2000); Aruga et al. (2002)
Thiothrix I and II Yes Yes Thiothrix spp. Howarth et al. (1999); Richard et al. (1982); Williams and Unz (1985a,
1985b, 1989); Tandoi et al. ( 1994); Kanagawa et al. (2000); Aruga et al.
(2002)
Type 0914 No No Unknown
,!
Beggiutoa Yes Beggiatoa spp. Williams and Unz (1985a, 1985b)
N. Iimicola I Yes Yes? Tricococcus spp. Liu et al. (2000,2002); Scheff et al. (1984)
N. Iimicola Il Yes Yes New genus, high GC van Veen(1973);Eikelboom (1975);Nowak and Brown (1990);Blackall
bacterium et al. (2000)
N. Iimicola II Yes Yes New genus, Snaidr et al. (2002\
Proteobacteria,
Alisphaeira europa
N. Iimicola II Yes Yes New genus,greensulfur Schade et al. (2002)
bacteria
N. Iimi<'ola lll Yes Yes? Isosphaera spp. Seviour.et al. (2002)
Type 0961 No No Unknown Richard et al. (1982\
Type 0092 No No Unknown Bradford et al. (1996)
Type 0581 No No Unknown
Type 0041 No Yes Member of TM7 group; Thomsen et al. (2002); Hugenholtz et al. (2001); Richard et al. (1982);
Bacterial domain Williams and Unz (1985a, 1985b)
Type 0675 No No New genus Hugenholtzet al. (2001)
Type l85l Ye s
,| New genus; Beer et al. (2002); Kohno et al. (2002); Bjomsson et al. (2002)
Chloroflexus group
Type 0803 Yes Yes New genus,p Blackall et al. (1996); Bradford et al. (1996);Williams and Unz (1985a)
Droteobacteria
M. parvicello Yes Yes New genus,Microthrix Slijkhuis (1983a, 1983b);Slijkhuis and Deinema(1982, 1988);Slijkhuis
parvicella et al. (1984); Blackall et al. (1994b, 1996);Rossettiet al. (1997a)
Nocardioforms Yes Yesfor Many genera Goodfellow et al. (1998); Soddell et al. (1998)
some
Type 1863 Yes Yes Acinetobacter spp., Rosetti et al. (1997'l; Seviour et al. (1997)
Moraxella spp.

Cur ds , l9' 15) . T o ta l n u mb e rs ra n g e fro m 100 to 2. MicroscopicEvaluation

>100,000/mL.Protozoaaregenerallydominant,with 500 to
To observethese organisms,place one drop (0.05 mL) of
severalthousand/ml observedcommonly.Theseorganisms
activated sludge on a microscope slide, add a cover slip,
perform severalimportant functions in activatedsludge,the
and examine it at l00x using phasecontrast illumination.
most important of which is the removal of nonflocculated
Count all protozoa and other higher life forms presentby
and loosely flocculatedbacteriafrom wastewaterto produce
scanning the entire cover slip area using the mechanical
a clarified effluent(Curdset al., 1968;Curdsand Fey, 1969).
stage of the microscope. Average the results of 4 to 5
Additionally, these organisms may contribute to biomass
separatepreparations.
flocculation through production of fecal pellets and mucus
(Curds, 1975) and may function to break up large floc Total number of organismspresent
massesand encourage a more active biomass through their per mL activated sludge culture =
motility (Javornicky and Prokesova, 1963). averagecount per cover glass areax 20
 Methods
FfGURE2.29 Phasecontrastmicrographsof:.(a) Flexibactersp., (b) Bacillus sp., (c) Oscillatoriasp. (cyanophyceae),
fungus.(Originalmagnification1000x.)(Seecolor insertfollowing page70.)
in dispersedform and were not distributed evenly throughout
the activated sludge floc. Rather they grew as dense col-
onies (Figure 2.30a) and the ammonia oxidizer colonies
were surrounded by the nitrite oxidizer colonies
(Figure 2.30b). These findings have significant implica-
tions, both for bioaugmentation and for the modeling of
nitrification in activated sludge.
The discrepancybetweenthesefindings and thosefrom
previous work in whichNitrosomonas andNitrobacter spp.
were identified as the principal nitrifying organisms in
activated sludge results from the bias introduced by cul-
turing microorganisms from environmental samples.
Because many environmental microorganisms will not
grow on laboratory culture media, the only way to be
certain whether an organism is important in an environment
such as activated sludge is to demonstrate its presence by
in situ methods that do not involve culturing and isolation.
An excellent example of mistaken identity causedby
the bias introduced by culture and isolation methods was
the conclusion that organisms of the Acinetobacter genus
were largely responsiblefor the phenomenonof enhanced
biological phosphorusremoval (EBPR) in activatedsludge
systems with initial anaerobic zones. Recent work has
clearly shown that this is not so. Microorganisms that are
not closely related to Acinetobacter spp. are important in
t-
this role (Hesselmannet al.. 1999: Crocetti et al.. 2000).
45
and (d)
D. Pnorozon Ano Mrrnzon
1. General
Microscopic observationof protozoa and other higher life
forms in activated sludge is a common and widespread
practice. In a very general way, the types of theseorganisms
present can be related to plant performance and efifluent
quality. They are useful for toxicity assessmentbut they
are of little or no value for determining the properties of
activated sludge that influence its behavior in solids sep-
aration processes.
From a morphological view, activated sludge is a rel-
atively simple microbial community consisting of free and
flocculated bacteria (and at times filamentous bacteria),
protozoa, rotifers, nematodes, and a few other inverte-
brates. Protozoa and other higher life forms are usually
aerobic and bacteriovorous (they eat bacteria). A few anaer-
obic flagellates and a number of saprophytic flagellates
occur in activated sludge. The saprophytic flagellates use
soluble organic matter for growth. Carnivorous free ciliates
and attached ciliates (suctorians) feed on other protozoa.
Chlorophyll-bearing flagellates are incidentally observed
and are usually derived from aeration basin walls.
Protozoa and other higher life forms may constitute
approximately 5%obyweightof the activatedsludgebiomass
and are represented by about 200 species (Curds, 1973;
Methods
FIGURE2.3O In silr,lidentificationof nitrifying bacteriain activatedsludge:(a) simultaneousin situhybidizationwith Cy3-labelled
probeNmV and FlUOS-labelledprobeNEU. Nitrosococcus
Nitrosococcusmobilis and Nitrosporalikebacteriaafterin situ hybidization with FlUOS-labelledprobeNmV (green)and Cy3-
labetledprobeS-*-Ntspa-1026-a-A18 (red).(FromJuretschko,
(Seecolor insertfollowing page70.)
If a large number of organisms are present (at times
observedfor flagellates),count the number presentin each
field of view (at 100x) for 10 to 20 fields of view, calculate
an average number per field of view, and multiply this
number by the number of flelds of view for the cover slip
area (this is typically about 300 at 100x magnification for
a 22 mm x 22 mm cover slip). Follow this procedure
frequently (several times a week or even daily) because
protozoan populations in activatedsludge can changerap-
idly in certain circumstances (e.g., an upset caused by
toxicity).
TaxonomicClassification
3.
Taxonomic classification of these organismsis basedpri-
marily on motility. The six basic groups observedin acti-
vated sludge are flagellates,amoebae,free-swimming cil-
iates, attached (stalked) ciliates, rotifers, and a few other
invertebrates.Identification to speciesis not necessary,but
recognition of the major groups of protozoaand higher life
forms can be useful in activatedsludgeoperation.The most
common protozoan and higher life forms observedin acti-
vated sludgeare shown in Figure 2.31 through Figure 2.33.
Thesephotos can be used as simplified identification keys.
More detailed taxonomic identification kevs can be
found as follows:
Protozoa Curds(1969and1975);Jahnetal. (1980);
MudrackandKunst(1986)
(1968);Doohan(1975);Gerardi(1987a)
Rotifers Calaway
Nematodes Calaway(1963);Schiemer(1975);Tarjan(1977);
Gerardi(1987b)
Annelids De L. G. Solbe(1975)
47
mobiliscells appearyellow; (b) simultaneous in situ identiflcationof
S. et al. (1998),Appl.Ewiron. Miuobiol.,64,3042.With permission.)
The major groups of protozoa and higher life forms
found in activated sludge are described below.
a. Flagellates
These are small (5 to 20 pm), oval or elongated forms
actively motile via one or more long, whip-like flagellae.
Many species found in activated sludge feed on soluble
organic matter and their presencecan indicate significant
soluble biochemical oxygen demand (BOD) levels. Many
of these occur at low dissolved oxygen (DO) and high
organic load.
b. Amoebae
Thesevary in shapeand size (10 to 200 pm) and are motile
via pseudopodia("false feet"). Some specieshave a hard,
ornate shell called a test (and the organisms are known as
testate amoebae, e.g., Arcella). Amoebae grow well on
particulate organic matter and are able to tolerate low DO
environments. A bloom of amoebaecan indicate a high
amount of particulate organic matter such as starch (pulp
and paper wastewater), yeast (brewery wastewater), and
septage(municipal wastewater).
c. Free-SwimmingCiliates
These are round to oval in shape (20 to 400 pm) and are
actively motile via rows of short, hair-like cilia. Some
species("crawlers") have cilia fused into spikes that aid
them in crawling on the surfacesof activated sludge flocs.
Ciliates are usually found under conditions of good floc
formation and generally indicate satisfactory activated
sludge operation. Ciliates are sensitiveand their presence
or absencecan indicate toxicity.
48 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FIGURE 2.31 Phase contrast micrographs of common flagellates and amoebaefound in activated sludge: (a) Monas sp., (b) Bodo
sp., (c) Polychaos sp., and (d) Arcella sp. (Original magnifications 1000x, (a) and (b); 400x, (c), and (d).) (See color insert following
page 70.)
d. Aftached Ciliates
These appearunder conditions similar to those producing
free-swimming ciliates but are found attachedto flocs by
stalks that may be either rigid or contractile. Some species
have one organism per stalk (e.g., Vorticel/c spp.) while
others are colonial (e.g., Epistylls spp. and Opercularia
spp.). Stalked ciliates generally occur at low organic load
(high MCRT). Individual speciescan be used as indicators
of approximateMCRT. The colonial forms occur at higher
MCRTs with more "heads" per colony as the MCRT
increases.
e. Rotifers
These have a variety of shapesand are much larger (50 to
500 pm) and have more complex structuresthan protozoa
(Figure 2.34). Most are motile and attach to activated
sludge flocs with contractile "feet." Theseorganismsoccur
over a wide range of MCRT; some speciesare indicative
of high MCRT.
f. Higher Invertebrates
These include nematodes(Figure 2.35d), tardigradessuch
as Macrobiotus sp. (Figure 2.35b), gasterotrichs
(Figure 2.35c), and annelids such as Nais sp. and Aeleo-
soma sp. (Figure 2.35a) which can impart a reddish color
to activated sludge due to their red- or orange-colored
"eyespots." Becauseof their low growth rates, nematodes
are generally observed only at higher MCRTs. Tardi-
grades,gasterotrichsand annelids appearto occur only in
nitrifying activated sludge systems,probably due to their
susceptibility to ammonia toxicity.
4. Use of Protozoa and Metazoa
as Indicator Organisms
Various protozoan and invertebrategroups develop in acti-
vated sludge according to growth conditions. Protozoa
have maximum growth rates of >l d-l at 20oC (Curds,
1975) and rotifers have growth rates of I to 2 d-l at20"C
(Doohan, 1975). Thus, the activated sludge growth rate
(or MCRT) rarely limits the development of these organ-
isms in most activated sludge systems.Nematodeshave a
lower maximum growth rate and generally develop only
in high MCRT systems (Water Pollution Control Federa-
tion, 1990). Food availability principally through freely
dispersedbacteria or turbidity is the primary determinant
of which group predominates.
Flagellates, amoebae,and small, free-swimming cili-
ates require high prey densities (>106to 107 bacteria/L)
becausetheir chase and capture feeding mechanisms are
Methods
FIGURE 2.32 Phaseconffast micrographs of free and stalked ciliates: (a) Aspidisca sp., (b) Paramecium sp., (c) Tokophyra sp., and
(d) Podophyra sp. (Original magnifications 400x, (a); 200x, (b), (c), and (d).) (See color insert following page 70.)
inefficient. These groups appear during plant startup and
at low MCRT (high organic load) conditions. Attached
ciliates, rotifers, and other invertebratesdevelop at lower
prey densitiesbecauseof their attachmentto the activated
sludge floc and their ability to feed by ciliary action (filter
feeding). These organism groups are selectedfor at high
MCRT (low organic load). These factors lead to marked
differences in the populations of various protozoa and
other higher life forms as activated sludge process oper-
ating parameters change (Table 2.20).
Satisfactory activated sludge performance occurs
when there is a balance among free-swimming and
attachedciliates and rotifers. An overabundanceof flagel-
lates, amoebae,or free-swimming ciliates is an indication
of high F iI\4 (low MCRT) while an overabundanceof
attached ciliates, rotifers, and other higher life forms,
especially nematodes,is an indication of low FnvI (high
MCRT). See Table 2.20. Because sludge settling often
deteriorates at organic loading extremes, many plants
attempt to adjust process parametersbased on the types
of protozoa and other higher life forms observed in the
activatedsludge.This is not a very sophisticatedapproach
to activated sludge settleability control, because many
other factors besides organic loading contribute to the
growth of the filamentous organisms that cause deteriora-
tion of activated sludse settlins.
49
One of the most valuable uses of microscopic observa-
tion of these organisms is toxicity assessment.These organ-
isms, particularly the ciliates and rotifers, are generally the
first to be impacted by toxic materials and can serve as ln
sira biomonitoring indicators for toxicants and other adverse
stresseson the activated sludge process.The first noticeable
sign of toxicity or stressis usually the slowing or cessation
of cilia movement in the ciliates. Next, the predominant
protozoan goups shift toward flagellates and small, free-
swimming ciliates that often "bloom" to high numbers
(>10,000/mL). This is an indication of activated sludge floc
breakup and the production of large numbers of dispersed
bacteria (turbidity) utilized as a food sourceby the flagellates
and free-swimming ciliates. Finally, in severecases,these
protozoa die, lyse, and releasetheir cell contents,sometimes
producing a white foam that contains dead protozoans and
protozoan fragments. Stressesother than toxicity that induce
these responsesinclude low DO, pH outside the range of
6.5 to 8.5, and high temperatures.Protozoa and other higher
life forms are generally absentfrom activatedsludge systems
operated at temperaturesabove 37 to 40"C.
E. Pnvsrcnl AND CHEMTCAT
METHoDs
In this section, we present specialized methods for mea-
suring the physical properties of activated sludge related
to solids separation.
50 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS IudgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Pr oblem s

FICURE 2.33 Phase contrast micrographsof stalked ciliates: (a) Opercularla sp., (b) Vaginicola sp., (c) Vorticella sp., and (d)
Epistylissp. (Original magnifications100x, (a); 200x, (b), (c), and (d).) (See color insert following page 70.)

FIGURE 2.34 Phasecontrastmicrographsof rotifers. (Original magnification100x.) (See color insert following page 70.)

1. SettlingTests 2. Foaming Tests

Three activated sludge settling tests are presented:(l) the
standard unstirred sludge volume index (SVI), (2) the
stined SVI at a standard initial SS concentration (SSVI
or SVIrr)(in this case,3.5 g SS/L), and (3) the diluted
SVI (DSVI). The relative merits of these methods are
discussed in Chapter 3. The SVI method is detailed in
Table 2.21, the SSVI method is found in Table 2.22, and
the DSVI techniqueis shown inTable 2.23.
Two foaming testsare presented.The first test (Table 2.24)
is suitable for activated sludge mixed liquor samplesand
aerobic and anaerobicdigestercontents.The Alka-SeltzeP
(Bayer Corporation, Morristown, NJ) test (Table 2.25) is
designed to determine the foaming potential of influent
and effluent streams.It is not suitablefor use with samples
containing high SS levels (such as mixed liquor and
digester contents).
Methods 51

FIGURE 2.35 Phasecontrast micrographs of invertebrates:(a) bristle worm(Aeleosoma sp.), (b) tardigrade (water bear, Macrobiotus
sp.), (c) gasterotrich(Chaetonotussp.), and (d) hydrachnidnematode.(Original magnifications100x, (a); 200x, (b) and (d); 400x,
(c).) (See color insert following page 70.)

TABTE2.20
Predominant
HigherLifeFormsObserved
at VariousActivatedSludgeOrganic
LoadingLevels
Condition PredominantGroups
Organic loading MCRT

High Low Flagellates, amoebae and small free-swimming ciliates

Moderate Moderate High diversity of organisms, dominated by free-swimming ciliates
Low High Stalked ciliates, rotifers, and higher invertebrates,especially nematodes

So u r ce s:F r o m Re yn o ld so n ,T .B.( 1 9 42),N ature,l 49,608;B ai nes,S .etal .(1953),

S ew agel ndustr.Wastes,
25,1923; Curds,C.R. and Cockbum,A. (1970a),Water Res.,4,225;Curds,C.R. and Cockbum,A. (1970b),
Water Res.,4, 237; Curds, C.R. (1975), in Ecological Aspects of UsedWater Treatment: The Organisms and
Their Ecology, Academic Press, NewYork, Chap.5; and Mudrack, K. and Kunst, S. (1986), Biology of
Sewage Treatment and Water Pollution Control, John Wiley & Sons, New York. With permission.

3. Methodsfor DifferentiatingMicrobiological 30 min in the container used to take the sample. This
and Process-Related
SolidsSeparationProblems avoids any changesin floc structure that could be caused
by sample transfers. This test indicates the state of floc-
The methodsdescribedin the following section were culation existing at the location in the treatmenttrain from
developedby Wahlberget al. (2001). which the sample is taken (Table 2.26).

a. Dispersed55 (DSS) b. Flocculated SS (F55)

DSS are the SS remainingin the supernatant
of an acti- FSS are the SS remaining in the supernatantof an activated
vatedsludgesamplethat hasbeensettledquiescentlyfor sludge sample that has been stirred with rotating paddles
52 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE2.21
StandardSludgeVolumeIndex(SVl)
Apparatus:
Graduatecylinder, I L.
Clock or stopwatchreadingin min.

Procedures:
1. Pour I L freshly-sampledmixed liquor into the graduatecylinder.
2. Allow to settlequiescentlyout of direct sunlight for 30 min.
3. After 30 min, record volume occupiedby the settledsludge.
4. Analyzea separatealiquot of the mixed liquor for total SS.

Calculation:

settledsludgevolume (mL/L) x t000

svr(-Lic)=ffi
Source:Ftom StandardMethodsfor the Exatninationof Waterand Waste'
water,?.Othed.(1998)AmericanPublic HealthAssociation,WaterEnviron-
ment Federation,and AmericanWater Worts Association,New York. With
permission.

TABLE2.22
of 3.5 g/t (SSVl3.s)
StirredSludgeVolumeIndexat SSConcentration
Settling apparatus:
Clear plastic (e.g.,Lucite@)cylinder, 100mm extcmal diameter,with a vertical scaleof 0 to 100mm, fitted
with a l2-volt, l-rpm DC motorand stirringdevice(Figure2'36).

Procedures:
1. DetermineTSS of mixedliquor sample.
2. If necessary,adjustmixedliquor TSS to 3.5 g/L by dilutionwith effluent(for samples>3.5 g TSS/L)
or by presettlingor mixing with return activatedsludge(for samples<3.5 g TSS/L).
3. Pour the 3.5 g TSS/L mixed liquor into the settling apparatusand immediatelystart t}le stirrer.
4. Allow to settle quiescentlyout of direct sunlight for 30 min.
5. After 30 min, record the volume occupiedby the settledsludge.

Calculation:

settledsludgevotume(m!L x 1OOO)
SS\1rs=ff

U.K. With permission.

Soarce..FromWhite, M.J.D. TechnicalReportTRl 1,WaterResearchCentre,Stevenage,

at 50 rpm for 30 min, thensettledquiescentlyfor 30 min c. SecondaryEffluentSS (ESS)

in the vesselin which it was stirred. FSS representsthe This is the SS concentrationin the secondaryclarifier -
secondaryeffluentSS that could ideally be achievedby effluent(Table2.2S).
secondarysettling of a well flocculatedmixed liquor -;
(Table2.27).
Methods 53

12 volt DC
Motor (1 rpm)

Removablelid

Drive shaft

Lucite@Tube

5 mm dia.
stirring rod

Basebearing

Base
-
FIGURE2.36 Apparatusfor SSVIr.r.(FromWhite, M.J.D. (1976),WaterPollut.Control,75, 459.With permission.)

TABTE 2.23
DilutedSVI(DSVI)Method
1. Set up severall-L graduatecylinders (the numberdependson prior knowledgeof the sludgesettleability).
2. Usingwellclarifiedsecondaryeffluent,prepareaseriesoftwo-folddilutionsofthemixedliquor(i.e.,nodilution, 1:ldilution,l:3dilution).
3. Stir the graduatecylindersindividually for 30 to 60 s, using a plunger to resuspendand uniformly distributethe SS.
4. Allow the activatedsludgeto settlefor 30 min underquiescentconditions.
5. Recordthe settledsludgevolume (SVr) in the first dilution where it is equal to or lessthan 200 mL (SV30<200 mL).
6. Calculate:DSVI (mug) = SV:o(mL/L)x 2"/TSS(g/L)

svr(d/L)x 2"
DSvr(mlls)
\ e/ =
' TS S (g/L)

wheren is the numberof two-fold dilutions requiredto obtain SV3o<20OmL andTSS is the total suspendedsolidsconcentrationof the
undiluted mixed liquor.

Source:From Stdbbe,G. (1964),Vertiffentlichungen

desInstitutesflir Siedlungwasserwirthschaft
der Technischen
HochschuleHannover,18.
With permission.
54 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE2.24
FoamingTestfor High SolidsSamples(Mixed Liquor and AnaerobicDigesterContents)
Reagents:
N, gas.
Ethanol957o w/v.

Foaming test apparatus:

Construct apparatusconsisting of a l-L graduate cylinder fitted at the center of its bottom with an air stone (ASTM 1745). Control the gas flow
to the air stone at 1600 cm3/min using an inlet valve, pressure gauge, and rotometer. Provide containment for the graduate cylinder to capture
material that can overflow it during the foaming test (Figure 2.37).

Procedures:
1. Adjust the temperature to 25"C if testing activated sludge and 37"C if testing anaerobically digesting sludge.
2. Adjust the TSS concentration to 2 g/L if testing activated sludge and to l.5Vo if testing anaerobically digesting sludge. Use distilled water
to a just TSS levels downward and settling to adjust TSS levels upward. Foaming tests can be conducted without adjustment of TSS
concentrations.
3. Gently pour 200 mL of sludge into a previously ethanol-washed and dried graduate cylinder. Aerate the sludge with N, gas at a rate of
1600 cm3/min for 90 s. When the foam layer reaches its maximum height, record the volume of the foam layer.

Source: From Hernandez, M.T. (1994), Ph.D. dissertation, University of Califomia, Berkeley. With permission.

Wooden
frame

Inlet

From N, gas
cylln o e r

FfCURE 2.37 Schematicdiagram of high solids foaming test apparatus.(From Hemandez,M.D. (1994), Ph.D. dissertation,Uni-
versity of California, Berkeley. With permission.)

TABTE2.25
FoamingTestfor Influent and EffluentStreams
Alka-Seltzer@
Reagent: Alka Seltzer@tablets (original unflavored, Bayer Corp., Morristown, NJ).

Equipment:
Graduate cylinder, 1 L (10 mL divisions).
Stopwatch reading in s.

Procedures:
1. Add 25 mL sample to a 500 mL graduate cylinder.
2. Drop in 2 Alka-Seltzer tablets.
3. Observe maximum foam volume.
4. When maximum foam volume is reached, start the stopwatch and record the time taken until the foam volume collapses so that
looking down the graduate cylinder from its top, 5OVoof the clear water surface is visible.
5. RepeatSteps I through 4 for tap water (control) and subtractthe control foam volume and collapse time values from the sample values.

Source: From Ho, C.F. and Jenkins,D. (1991), Water Sci. Technol.,23:44,879. With permission.
Methods JJ

TABTE2.26
DispersedSS(DSS)Method
1. Collect samplesusing a 4.2-L acrylic Kemmerer sampler (Model
1540-C20,Wildlife Supply Co., Saginaw, MI, Figure 2.38). The
sampler is a clear tube, 105 mm (4.1 in.) in diameter and 600
mm (24 in.) tall with upper and lower closures.Lock the closures
in the open position.
2. Lower the sampler to the desired depth with a rope. Close the
closures by dropping a weighted messengerdown the rope.
3. Quickly raise the sampler and immediately open the bottom drain
valve to lower the liquid level in the samplerjust below its upper
internal support.
4. Secure the sampler in a vertical position, open the upper closure,
and start a stopwatch.
5. After approximately 10 min of settling, gently twist the upper
closure to dislodge any SS that may have adhered to its straps.
6. After 20 min of settling, immerse one end of a primed siphon
into the sampler to a depth that will allow the withdrawal of
slightly more than 500 mL of supematant.
7. After 30 min of settling, open the siphon and allow approximately
50 mL of supernatantto flow to waste.
8. After the siphon has cleared, collect a 500-mL sample. Use low
supematantwithdrawal rates (approximately 150 to 200 ml/min)
to prevent disruption of the sludge blanket. To prevent the inclu-
sion of floating solids, stop supernatant withdrawal when the
water surface falls within approximately 6 mrn (0.25 in.) of the
siphon inlet. Analyze the supernatantfor TSS.
TABLE2.27
FlocculatedSS(FSS)Method
1. Use a 6-place paddle stirrer (Phipps and Bird, Richmond, VA).
2. From the location in the flow scheme to be investigated, collect
at least 1.5 L of mixed liquor.
3. Place the flocculation jar on the paddle stirrer and mix 30 min
at 50 rpm.
4. Allow the sample to settle for 30 min.
5. Take a supematant sample using a siphon device or a tube
attached to a vacuum source.
6. Carefully avoid introducing settled SS or floating SS into the
supernatantsample.
7. Analyze the supernatantsample for TSS.
Source: From Wahlberg, E.J. and Keinath, T.M. (1995), Water Environ
Res.,67,872. With permission.
TABLE2.28
EffluentSS(ESS)Method
l. Collect an effluent sample from the secondary clarifier at the
same time the DSS and FSS tests are conducted.
2. Analyze secondary effluent sample for TSS.

Source: From Wahlberg, E.J. and Keinath, T.M. (1995), Water Environ. Soarce.'From Wahlberg,E.J. and Keinath,T.M. (1995), WaterEnviron.
Res.,6'1,872. With permission. Res.,6i ,872. With permission.

Upper internal Upper closure seal

Water level before
supernatantsampling
Water level after
Sampling port

Clear supernatant

Settled sludge

Lower internal supporl

Lower closure seal

Bottom drain valve

FIGURE 2.38 Schematicdiagram of Kemmerer sampler in use. (From Wahl-

berg, E.J. (2001), Final Report for Project 00-CTS-1, Water Environment
ResearchFoundation, Alexandria, VA. With permission.)
 ? Applicationsand Results
\-'
of MicroscopicExamination
of ActivatedSludg"
Where the telescopeends,the microscopebegins.Which
has the sranderview?
Victor Hugo (1802-1855),St.Denis,Book 1
I. INTRODUCTION
This chapterdiscussesthe type of information that can be
obtained from a microscopic examination of activated
sludge.Applications of this information to the diagnosis
and resolutionof activatedsludgesolids separationprob-
lems are presented.A discussionof the use of protozoa
and higher life form characterizationin activated sludge
was presentedin Chapter2. The methodsused to differ-
entiatebetweenmicrobial and process-related solids sep-
arationproblemsare presentedin Chapter4. A discussion
of the useof microscopicmethodsand testingprocedures
relatedto foaming is reservedfor Chapter5.
II. FITA ME NT
COUNTING
The counting of the number of filamentsextendingfrom
activated sludge flocs or the measurementof the total
extendedfilamentousorganismlength was used by early
workers to demonstratethat the settling properties of an
activatedsludgecould be relatedto its filamentousorgan-
ism level.
Finstein and Heukelekian (1967) showed that SVI
could be relatedto the total Iilament lengthper floc. Pipes
(1979) counted the number of filamentous organisms
extending from activated sludge flocs and found that, at
low filament numbers(<102filaments/mgVSS), the SVI
was below 100 ml-/g, while at high filament numbers
(>102to 103filaments/mgVSS), the SVI increasedsignif-
icantly.
Sezginet al. (1978),Palm et al. (1980),and Lee et al.
(1982) all used the Sezgin method (Table 2.1) for mea-
suring the total extended filament length (TEFL) in acti-
vated sludge. All these investigators found correlations
between activated sludge settling measurements(such as
SVI and zone settling velocity) and TEFL for laboratory-
scale activated sludge grown on domestic wastewater.In
general,the SVI increasedrapidly above 150 mllg when
TEFL values increasedabove 107 pm/ml (Figure 3.1).
Sezgin et al. (1980) found that these relationshipswere
valid for activated sludse taken from several full-scale
plants (Figure 3.2).
It is soberingto think about the filamentousorganism
level neededto causean activatedsludgesettlingproblem.
SVI values increaseabove 150 ml-/g when TEFL levels
exceed about 107 intersections/ml. This means that it
takes more than 10 m (about 33 ft) of filamentsin each
mL of mixed liquor to causea settlingproblem.Lee et al.
(1983) determined the relationship between TEFL and
sludge settleabilitymeasuredby severaltechniques:SVI
and dilutedSVI or DSVI (St6bbe,1964),SVI at SS con-
centrationsof 1.5, 2.5, and 3.5 glL, and stirredSVI at 2.5
g SS/L (similar to the SSVI.,rof White, 1976).Lee and
coworkersreasonedthat becauseTEFL was a quantitative
index of sludge settleability,the settling test to which it
best correlatedwould be the best one for judging sludge
settleability.Becausethe closestcorrelationwas obtained
between TEFL and DSVI, they proposedreplacing the
standardSVI with the DSVI.
Similar conclusionswere reachedby Matsui and
Yamamoto(1983),who useda video imaging systemto
aid in the microscopiccountingof filamentlength.Further
justification for this proposalwas provided by Koopman
and Cadee (1983), Pitman (1985), and Rachwal et al.
(1982) who showedthat the parametersrelating activated
sludge settling velocity and activatedsludge SS concen-
tration correlatedwell with the DSVI. Thus, the DSVI test
is valuable for predicting activated sludge separation
behavior in process units and correlateswell with the
activatedsludgefilamentousorganismlevel.
Simplified methods for determining filamentous
organismlevels in activatedsludge yield results that are
well correlatedwith settling properties.Thus, the subjec-
tive scoring systemfor filamentousorganismabundance
ffable 2.15) was usedin an industrialwastewateractivated
sludge plant and the results correlated well with SVI val-
ues measuredby an independentlaboratory(Figure 3.3).
Data obtainedfrom the SanJose/Santa Clara (Califor-
nia) Water Pollution Control Plant (SJ/SCWPCP) using
the simplified filament count method (Table2.2) to assess
the filamentousorganismlevelsin activatedsludgeillustrate
J/
5B Pr oblem s
M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on

800 6 Excessive

700 5 Abundant
2N
il

) ooo n Yery
{9 1s0 common
f .no ,i
E
t.:" 3 Common
F-i
9 400 ; 100 2 Some
I roo 50
il I Few
E zoo
(h
0 0 None
1 00 Junl I Jul 31 S epl 9 N ov8 D ec28 Febl 6
Date.1991 - 1992
0
lo5 106 107 108
ExtendedFilamentLensthulrl/ml-
FIGURE3.3 Relationshipof subjectivescoringof filament
abundanceand SVI as measuredby independentobservers.
FIGURE 3.1 Effect of extendedfilament length on SVI. (From (FromBeebe,R.D. et al. (1982),paperpresented
at 55thannual
Palm, J.C. et al. ( I 980), J. WaterPollut. Contol Fed., 52, 2484. of theWaterPollutionControlFederation.
conference St. Louis.
With permission.) MO. With permission.)

which a given number of filamentous organisms affect

1400
settleability.
The observationof Lee et al. (1983) that TEFL and
DSVI were well correlated and the finding of Beebe et al.
1200
(1982) that an increasein filament count did not precede
Curve determined from
laboratory studies
a decreaseof settleability means that filament counting
1000
of activated sludge is not an efficient process control
method for sludge settling. It is far easierto do a DSVI
{ 8oo
testthanto count filaments.Sinceboth testsyield the same
j information at the sameinstantin time, it makessenseto
e 600 do the easier test. However, filament identification is an
extremely useful tool for diagnosing and rectifying
activated sludge solids separation problems.

200
I I I . F I L A ME NT O US
O RCA NI S M
0 IDENTIFICATION IN ACTIVATED STUDGE
1 05 100 107 l 0o
pm/mL A. Rrsulrs oF FTLAMENTouS SuRVEys
ORcANTsM

FIGURE3.2 Variationof SVI with filamentlengthfor 14 full- The filamentous organism morphotypes observed in acti-
scaleCalifomiawastewater treatment plants.(FromSezgin,M. et vated sludgewere investigatedin a systematicfashionby
al.(1980),"/.WaterPollut.ControlFed.,12,171.Withpermission.) many workers including Cyrus and Sladka (1970), Hi.iner-
berg et al. (1970), Farquhar and Boyle (1971a, l97lb),
two important points (Figure 3.4). First, because changes Sladkaand Ottova(1973),Eikelboom(1975,1977),Elkel-
in SVI and filament count occur at the same time, the boom and van Buijsen (1981),Richardet al. (1982),Wag-
filament count cannot be used as an early warning for an ner (1982), Strom and Jenkins (1984), Blackbeardet al.
increasein SVI (Beebe et al., 1982).Second,when these (1986), Richard (1989), Seviouret al. (1994), Kristensen
measurements were made,the plant consistedof two acti- et al. (1994), Rossetti et al. (1994), and Wanner et al.
vated sludgeplants in series.The first (or secondary plant) (1998).
was for carbonaceousBOD removal and the second (or Their studies establishedthat 20 to 25 or more differ-
tertiary plant) was for nitrification. The filamentous organ- ent morphological types of filamentousmicroorganisms
isms found in theseplants were different and, as Figure 3.5 can be found in activated sludges treating municipal
shows,SVI and filament count had a different relationship. wastewaters.Eikelboom and Geurkink (2001) suggested
This suggests that the growth form of the filamentous that activated sludgestreating industrial wastewatersmay
organism in activated sludge and therefore the type of contain considerably more morphological types of fila-
filamentous organismspresentcan influence the degreeto mentousorsanisms.
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge 59

200

-! 180
160
(,3
tr
k 120
a=
F roo
E
d80
b!

?6n

U)

10 20 30 10 20 30 10 20
Mar Apr May
Date.1982

FIGURE 3.4 Comparisonof filament count and SVI at San Jose/SantaClara plant in California. (From Beebe,R.D. et al. (1982),
paper presentedat 55th annual conference of the Water Pollution Control Federation, St. Louis, MO. With permission.)

varies significantly. The differences can be explained by

variationsin wastewatertype and strength;plant operating
conditions (particularly FA{, MCRT, DO concentration,
120 and temperature);and aeration basin design (especially
initial mixing conditionsbetweenthe influent wastewater
and the RAS and the presenceof initial anaerobic,anoxic,
or otherwiseunaeratedzonesin the aerationbasin).
100
I

Fi
B. Drncnosrs oF CAUsEsoF SouDs SrpnnnrroN
u) 80 PnosLrlisrHRoucH MrcnoscoprcExniurNnrroru

Second stage
20
0 20 40 60 80
"Filament
Count"
FIGU RE3.5 Relationshipsof filamentcountto SVI for firstand
secondstageactivatedsludgeplantsat the SanJose/Santa Clara
plantin California.
Surveys of the appearancesof various filamentous
organism types in bulking and foaming activated sludge
were conductedin the U.S. (Richard et al., 1982; Strom
and Jenkins, 1984; Richard, 1986), Northern Europe
(Eikelboom, 1977 ; W agner,1982; Kristensenet al., 1994),
South Africa (Blackbeard et al., 1986a,b),Australia
(Seviouret al.,1994); Italy (Rossettiet a1.,1994),and the
Czech Republic (Wanneret al., 1998),SeeTable 3.1 and
Table 3.2. These surveys demonstratedthat the same fil-
amentous organism types are observed ubiquitously in
activated sludge and that approximately l0 to 12 types
account for the great majority of bulking and foaming
episodes.The relative frequency of occurrence of domi-
nant individual filamentous organisms in bulking sludge
1. C eneral
Much can be learnedfrom a careful microscopicexami-
nation about how an activatedsludgewill behaveduring
solids separationprocesses,especiallyif the observeris
alert and keeps an open mind - some strangethings are
100 found in activated sludge. A microscopic examination
should always be conductedwhen addressingsolids sep-
arationproblems.The data producedfrom a microscopic
examinationarecompletelyindependentof other analyses
done with an aim to resolving such problems. Further-
more, microscopic observation of the flocs and filamen-
tous organisms provides a measurementthat reflects not
only the current conditions, but to some degreeconditions
that have existed over at least the previous one or two
MCRTs.
Microscopic observation of activated sludge has its
greatest value when used in conjunction with data from
chemical analyses,operating records, and plant design.
Becausemicroscopicobservationstend to be subjective,
it is not a good idea to learn a lot about an unknown sample
before looking at it. If someonetells you what he or she
thinks the problem is, it is easy to convince yourself that
you seewhat you think you should see.
This section details information that can be gleaned
from determining the types of filamentous organisms
60 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE3.1
in U.S.Bulkingand FoamingActivated
FilamentAbundance
SludgePlants
Percentageof Treatment
Plantswith Bulkingor
Foamingin Which
FilamentsWere:
Rank FilamentousOrganism Dominant Secondary

I Nocardioformorganisms 31 17
2 Type 11O1 29 24
3 Type021N 19 15
4 Type0041 16 4'7
5 Thiothrixspp. 12 20
6 Sphaerotilusnatans 12 19
'l Microthrixpanicella 10 3
8 Type0092 9 4
9 Haliscomenobacter hydrossis 9 45
l0 Type0675 '7 16
I1 Tlpe 0803 6 9
12 Nostocoidalimicola (typesI, II and III) 6 18
13 Tlpe 1851 6 2
14 Type0961 4 6
15 'Ilpe 0581 3 1
16 Beggiatoaspp. | 4
17 Fungi | 2
l8 Type0914 I I
All others I -

Note:Combined resultsofRichardet al. (1982)andStromandJenkins(1984);

525 samplesfrom 270 treatmentplants.

present.While this is very useful in problem diagnosis, it
is not the only type of thing that can be learned from
microscopic examination of activatedsludge.Much useful
information can be obtained from observing floc charac-
teristics and the presenceof other types of organisms or
nonbiological particles.
2. Nonmicrobial partictes
The nonmicrobial particles present in activated sludge
flocs can be informative. Most activated sludge contains
paper fibers, most likely from toilet tissue. Pulp and paper
wastewateractivatedsludgealways containsfibers derived
from the pulped material. The observation of many paper
fibers (Figure 3.6a) in municipal wastewater activated
sludge suggeststhat the plant does not have primary clar-
ifiers becausemost paper fibers settle out in primary clar-
ifiers. Also seen in the activated sludge from this type of
plant are hairs (Figure 3.6b), plant ducts (Figure 3.6c),
and greaseparticles (Figure 3.6d). The presenceof many
paper fibers in the activated sludge from a plant with
primary clarifiers is an indication that the primary clarifi-
ers function inefficiently.
Finding large numbersof amorphousorganic particles
(Figure 3.7a) in municipal wastewater activated sludge
suggeststhe presenceof recycled digested sludge solids
somewherein the solids handling processes.Examples are
digester supernatant,thickener return flows, centrate, fil-
trate, etc. In severecasesof solids recycling, the odor of
digested sludge will be apparent in the activated sludge
sample. Often this observation is accompaniedby the
finding of low percentVSS in the mixed liquor SS, turbid
secondaryeffluent, and a pumice-like foam on the aeration
basin and secondaryclarifiers.
Hydrogen sulfide reactswith iron saltspresentin most
wastewatersto form a black precipitate of iron sulfide that
accumulatesin the floc (Figure 3.7b). The finding of sig-
nificant iron sulfide is an indication of septic wastewater
or high levels of hydrogen sulfide somewherein the sys-
tem. Often, this is a sign of septic return flows to the
aeration basin, usually from a solids handling process.
When iron salts are added to the primary effluent or
to the mixed liquor (for example, for phosphateremoval
by simultaneousprecipitation), it is usually possible to see
yellow amorphous areas in the flocs consisting of ferric
phosphate and hydroxide precipitates (Figure 3.7c).
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge 61

TABTE3.2
Comparisonof Dominant FilamentousOrganismsin Bulkingand FoamingActivatedSludge
from SeveralCountries
Rankingin Order of Prevalence
The
Filamentous Organism U.S." Netherlandsb," Germanyc," South Africad Australiar Denmarke Italyh CzechRepublici

Nocardioforms and I
,| 3 26
actlnomycetes
Type 1701 2 588 t2 9 13
Type 02lN 3 2l I 3 67
Type 0041 636 2 2 35
Thiothrix spp. 199 l3 78
S. natans 6 74 T4
M. panicella 7 t 22 I I 1t
Type 0092 8 4- 1 4 3 +)
H. hydrossis 9 369 5 1l
Type 0675 l0 A 2 3 11
Type 0803 ll 9107 o 4
N. limicola I, II, and III l2 11 7 8 5 52
Type 1851 l3 123 8 ;
Type 0961 l4 109- 10 89
Type 0581 l5 8- 9
Beggiatoa spp. l6 l8 l)

Fungi t7 15;
Type 0914 t8 , ; l0 l0

" Richard et al. (1982) and Strom and Jenkins(1984):525 samplesfrom 270 plants.
b Eikelboom (1977): 1100 samplesfrom 200 plants.

" Wagner (1982): 3500 samplesfrom 315 plants.

d Blackbeardet al. (1986a and 1986b): I I I plants including 26 BNR plants; bulking and nonbulking sludge.

" Nocardioforms not included.

r Seviour et al. (1994): 65 samplesfrom 65 plants.
c Kristenesenet al. (1994): 38 EBPR plants.
h Rossettiet al. (1994): 40 plants.
i Wanner et al. (1998): 86 samplesfrom 86 plants.

Because phase contrast illumination distorts color, it is
necessaryto confirm the true color ofthese areasby view-
ing them under direct illumination. Inorganic precipitates
in activated sludge also reduce its volatile fraction.
In the powdered activated carbon treatment (PACT)
process (Hutton and Robertaccio, 1975), powdered acti-
vated carbon is added to the activated sludge. It is easy to
see the black angular particles of powdered activatedcar-
bon microscopically in PACT-activatedsludge even at a
magnification of 100x (Figure 3.7d). The carbon particles
are thick and strong enoughto prevent one from preparing
a very thin preparation by pressing on a cover slip; at
1000x magnification, it is difficult to focus properly. The
presenceof isolated carbon particles in activated sludge
treating industrial wastewatersusually meansthat the water
used for quenching or scrubbing flue gases is being dis-
charged into the wastewatertreatment plant.
Many other nonmicrobial particles are found in acti-
vated sludge treating industrial wastewaters. These
include dye particles, resin beads, diatomaceous earth
(Figure 3.8a), catalyst support particles, oil and grease
droplets (Figure 3.8b), fibers from clothing manufacture
(Figure 3.8c), and starchgranules(Figure 3.8d).
3. Other Microbiological Features
a. General
Microscopic observation of the condition of activated
sludge flocs and the occurrence of microorganisms other
than filaments can help judge the nature of the growth
environment in the aeration basin. Important floc charac-
teristics are size, strength, apparentbacterial morpholog-
ical diversity, specific morphological types of bacteria
indicative of growth conditions, dispersedgrowth, and the
presenceof algae, protozoa, and metazoa.
b. Limited Diversity
In a typical activated sludge floc developedon municipal
wastewater, one usually observes bacteria with a wide
variety of morphological forms, suggesting the presence
62 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FIGURE3.5 Phasecontrastmicrographs of nonmicrobialparticles:(a) pap'erfibers,(b) hair,(c) plantduct,and(d) grease.(Original
100x,(a) and(b);200x,(c) and(d).)
magnifications
of a number of bacterial types growing on a mixture of
substrates(Figure 3.9a). Activated sludge flocs developed
on industrial wastewatersoften exhibit limited diversity
of morphological types of bacteria, indicative of growth
on a wastewaterwith a simpler composition than munic-
ipal wastewater (Figure 3.9b and Figure 2.7). Such acti-
vated sludges may not be as resilient to changesin envi-
ronmental or wastewaterconditions as those developedon
a more diverse substrate.
Often industrial wastewatersystemsexperience short
periods of dispersed bacterial growth and high effluent
turbidity when the wastewatercomposition or temperature
changes. This may be due to a change in predominant
floc-forming species.
c. Dispersed Growth
For an activated sludge to produce a well clarified, low
TSS effluent. it must contain low levels of individual bac-
terial cells and small cell aggregates(both termed dis-
persed growth). Large amounts of dispersed growth
(Figure 1.5) in the aeration basin will produce a turbid
effluent. Dispersed growth is encounteredunder a variety
of conditions. The diagnostic methods of Wahlberg et al.
(2001) are very useful in determining which of these sev-
eral conditions is causing the dispersedgrowth.
An activated sludge sample stored in a full bottle,
especially if it has become warm and septic, will develop
dispersed growth. This is an artifact of improper sample
collection and storage; it occurs more rapidly with low
MCRT (poorly stabilized) activatedsludge than with high
MCRT (well stabilized) activatedsludge.A fresh, properly
collected and stored sample should always be examined.
High F/TvI(low MCRT) increasesthe amount of dis-
persed bacteria in activated sludge. In municipal waste-
water activated sludge, dispersedgrowth can start to exert
a noticeableimpact on effluent quality at MCRTs less than
2to3d.
For activated sludge developed on high organic
strength, readily biodegradable,soluble industrial waste-
waters, dispersed growth can occur at much lower F/IVI
values. Spills and shock loads in industrial wastewater
activated sludge systems typically result in dispersed
growth and turbid effluents. Dispersedgrowth is common
in trickling filter and RBC effluents, especially from high
rate units and causes the turbidiry often seen in trickling
filter effluents. Dispersed growth arisesbecauseof inade-
quate bioflocculation time afforded by high rate filters. The
dispersed microorganisms can be removed by biofloccu-
lating the filter effluent using processessuch as the trickling
fi lter/solids contact (TF/SC) process(Nonis et al., 1982).
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge 63

FICURE 3.7 Phase contrast micrographs of nonmicrobial particles: (a) a.rnorphousorganic matter, (b) iron sulfide, (c) ferric iron
precipitate,and (d) activatedcarbon.(Original magnifications200x, (a) and (b); 1000x, (c); 100x, (d).) (See color insert following
page 70.)

Novak and coworkers ( Higgins and Novak, 1997a,
1997b; Novak, 2001; Murthy and Novak, 1998, 2001)
have shown that, under some circumstances,dispersed
growth in activated sludge can be caused by what they
term a cation imbalalance - the relative concentrations
in the wastewater of monovalent (Na*, K*, NHo*) and
divalent (Ca2* and Mg2*) cations. When the ratio of
monovalent cations to divalent cations is high, dispersed
growth can occur.Novak and his group suggestthat micro-
organismsin activatedsludge attach to each other through
the bridging of their exocellular polymers (polysaccha-
rides and proteins)(seeFigure 1.1). Divalent cationspar-
ticipate in this bridging while monovalent cations do not.
Besides influencing the amount of dispersed growth in
activated sludge, Higgins and Novak (1997a) showed that
wastewater cation levels also affect settling rates, resis-
tance to filtration, and thickness of cake produced by
various solids' thickening processes.
The two causesof dispersedgrowth discussedabove
result from inadequatebioflocculation and poor floc for-
mation. Dispersedgrowth can also result from the breakup
or deflocculation of existing flocs. Three generic causes
for this type of dispersed growth have been identified:
mechanical shearing,the presenceof surfactants,and the
action of toxicants. The mechanical shearing or breakup
of flocs occurs in activated sludge systemswith vigorous
methods of aeration (e.g., mechanical aeratorsand coarse
bubble diffused air). This type of floc is referred to as pin
or pinpoint floc. In such sludges,the majority of the bio-
mass,with the exceptionof the small dispersedfloc, usu-
ally settles rapidly. Floc shearing to produce dispersed
growth can result from other types of rough treatment of
the activated sludge, such as falling over weirs and into
drop structures; pumping, especially with a centrifugal
pump; and flow through long and tortuous piping systems.
Pin floc can be rectified by reflocculating small flocs
with larger ones. This can be done by ensuring gentle
transfer of mixed liquor between the aeration basin and
the secondary clarifier and by providing a gently mixed
flocculation zone in the secondaryclarifier. To achievethis
in a circular secondaryclarifier, one should provide alarge
tangentially fed center well with sufficient hydraulic
detention time to provide for reflocculation of the small
flocs (Wahlberg et al., 1994).
Surfactants can cause dispersed growth through floc
dispersion. For this effect to be manifested,the surfactant
must be present in the aeration basin and must be slowly
biodesradable or nonbiodesradable. The most common
64 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

FIGURE 3.8 Phasecontrastmicrographsof nonmicrobial particles:(a) diatomaceousearth, (b) oil droplets,(c) clothing fiber, and
(d) starchgrain. (Original magnifications200x, (a) and (d); 100x (c);400x (b).)

FIGURE3.9 Phasecontrastmicrographs diversity of activatedsludgeflocs: (a) high diversity,(b) low

showingmorphological
diversity.(Originalmagnification1000x.)

biodegradable surfactants encountered in U.S. wastewa-
ters are the branchedchain, alkyl phenol ethoxylate (APE)
or nonyl phenol ethoxylate (NPE) nonionic surfactants.
The floc dispersion effect is also noticeable in the
behavior of the secondaryclarifier sludge blanket which,
insteadof possessinga distinct interfacebetweenthe set-
tled sludge and the supernatantliquid, becomes diffuse,
showing a very gradual transition from the sludge layer
to the supernatantover a distance up to several feet. An
analysisfor surfactants(anionic and nonionic) and surface
tension should be made for confirmation. Surface tension
values below 60 dynes/cm2in the aeration basin liquid or
in the secondaryeffluent are indicative of such a problem.
The presenceof white, frothy foam on the surface of the
treatment unit should also raise suspicionsthat surfactants
may be present.We have seenthis type of surfactant floc
dispersion problem in activated sludge treating surfactant
wastewater,pharmaceutical wastewater,textile waste-
water, commercial laundry wastewater,and occasionally
in wastewaters from industries that engage in periodic
equipment cleaning, e.g., dairies, cheese manufacturers,
and canneries.This type of floc dispersion problem may
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge
be seasonal.It tends to be worse when the wastewateris
colder because the biodegradation rates of some of the
causative surfactants decrease at low temperatures
(Kravetzet al.,1984).
The presence of toxicants (especially heavy metals)
in influent wastewatercan result in dispersedgrowth due
to deflocculation. Filamentous organisms,if presentin the
activated sludge, are often the first microorganisms to be
affected by toxic metals.The SVI decreasesrather rapidly
and freely dispersed,damagedfilaments can be observed
microscopically in the supernatant above the settled
sludge.Ifthe toxicity is severeor frequentenough,defloc-
culation will occur. Bott and Love Q002\ and Wimmer
et al. (2001) have shown that deflocculationdue to elec-
trophilic toxicants (e.g., heavy metals) can be preceded
by a releaseof potassiumions to yield K* concentrations
of 2 to 5 g/L inside the flocs.
Since Murthy and Novak (1998) estimatethat K+ lev-
els of approximately0.7 g/l- will causefloc dispersion,it
is possible that the dispersion associatedwith toxicity
could in fact be due to the K* ion releasethat the toxicant
elicits. Following deflocculation,protozoa,usually flagel-
lates,often "bloom" due to the suddenavailabilityof food
sources(deflocculatedbacteria).If the toxicity event is
severeenough,these protozoaare killed and their lysed
cell contentscan producea foam. Microscopic examina-
tion of this foam will reveal the presenceof dead proto-
zoansandprotozoanfragments.The activatedsludgeBOD
removal usually declinesor ceasesfollowing this type of
event. Activated sludge affected by severeheavy metal
toxicity will have a much higher than normal heavy metal
contentin the MLSS. This can be determinedby chemical
analysis.
A similar sequenceof eventscan occur during RAS
chlorination for bulking control (see Chapter 4). If the
initial mixing of chlorine and RAS is poor or if the chlo-
rine dose is too high, the floc breaks up, forming small
floc fragments. Dispersed growth and turbid effluents
result when chlorine dosing is very high.
Aeration basin temperaturesabove 35 to 40'C can
often causedispersedgrowth of floc-forming and filamen-
tous organisms(Norris et al., 2000; Parkset al., 2000)This
is an increasingproblem in many industrial wastewater
treatment plants in which water conservation practices
reduce effluent volume without reducing process heat
losses, thereby increasing wastewater temperatures.A
common observationis the occurrenceof episodesof dis-
persedgrowth of single bacteriaand dispersedfilaments,
high effluent turbidity, and loss of floc strength as the
aeration basin temperatureincreasesfrom below 35oC to
above this value. The dispersed growth and effluent tur-
bidity often subsideafter a few days as new thermotolerant
floc-forming bacteria develop. Dispersedgrowth episodes
occur also as the temperaturedecreasesthrough this range,
perhapsbecausethe thermotolerant floc formers wash out
65
of the system as they are replaced by mesophilic floc
formers. For this reason,in activatedsludge systemsoper-
ated at high temperatures(>35oC), it is important to limit
temperaturevariations as much as possible.
Dispersed growth and turbid effluents can be caused
by the growth of severaltypes of microorganisms free in
suspensionand not attachedto the flocs.Theseorganisms
can be filamentous (e.g., H. hydrossis, Type 0914,
N. limicola, Type 1863, and nocardioformorganisms)or
one of several types of single-celled bacteria of unique
appearance.One of the more common of theseis a Gram-
negativeand Neisser-positivestaininggroup offour cells,
termed the Neisser-positive tetrad (Figure 3.10a and
Figure 3.10b). This organismhas been observedin asso-
ciation with nutrient deficiency and the presenceof low
molecular weight organic acids in the wastewater.
d. Neisser-PositiveCell Clumps
Neisser-positive staining bacteria that grow in clumps of
often grape-likeclustersinside the activatedsludge flocs
appearin systemswith initial anoxic or anaerobiczones
(selectors)in the aeration basin (Figure 3.10c). The
Neisser-positivestainingreactionis thought to be associ-
ated with internally storedinorganicpolyphosphategran-
ules that accumulatewhen these organismsare cycled
through an anaerobic/aerobicenvironment.These types of
Neisser-positivestaining cells are found in significant
numbers in activated sludge systems exhibiting EBPR
(e.g., processessuch as Bardenpho,A/O, Phostrip, and
UCT). Whife originally thought to be Acinetobacterspp.,
it hasnow beenshownthat someof thesemicroorganisms
are membersof the genus RfutdocycLus(Crocetti et al.,
2000; Hesselmanet al., 1999).
In addition to clumps of microorganismsthat stain
evenly Neisser-positive,Neisserstaining will sometimes
also reveal clumps of cells that stain strongly positive
(deep blue-purple)in and near their cell walls and less
strongly positive (light blue) in their interiors (Figure
3.10d). Theseare refened to as G bacteria(Cech and Hart-
man, 1993)and are associatedwith activatedsludgesystems
that have initial anaerobic/anoxiczones in which EBPR
does not occur. These organismsapparentlycan take up
and store soluble substrateanaerobically/anoxicallywith-
out the accompanying cycling of phosphate and storage
of inorganic polyphosphate.They are sometimesfound in
EBPR-activated sludge systems in which EBPR has
ceased to function or functions poorly. They were first
observedby Cech and Hartmann (1993) in laboratory-
scale anaerobic/aerobic activated sludge units fed with
glucose (hence the name). We have seen G bacteria in
large numbers in a full-scale anaerobic/aerobicactivated
sludge unit treating wastewater from a sugar refinery.
Mazemannet al. (1991, 1999) assignedsome G bacteria
to the new generaAmaricoccus and Tesssaracoccus.
66 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

FIGURE 3.10 Neisser-positivecell clumps in activated sludge flocs: (a)'tetrad, phase contrast of wet mount; (b) tetrad, direct
illumination Neisser stain; (c) phosphate-accumulatingorganisms (PAOs), direct illumination Neisser stain; and (d) G bacteria, direct
illumination Neisserstain. (Original magnification1000x.) (See color insert following page 70.)

e. Yeast ifiers often accompanies the observation of high yeast

The presenceof large amountsof yeastcells (Figure 3. 1la)
in activated sludge is unusual and suggestsone of four
conditions:
L Yeastin the influent wastewater(e.g., wastewa-
ters from bakery, brewery, or other industrial
fermentation processes)
2. Fermentative conditions somewherein the col-
lection system or wastewatertreatment plant
3. Low pH (<6.0) levelssimilar to thosethat cause
fungi (to which yeasts are related)
4. SevereP deficiency
We have seen yeast proliferate in activated sludge in
a systemaeratedwith fine pore diffusers when the diffusers
became clogged and the oxygen supply was insufficient.
Another yeast overgrowth incident was associatedwith
the fermentation of a sugar-containing wastewater and
domestic wastewater mixture in the primary clarifiers of
the plant. Septic conditions in raw sludge gravity thick-
enerscan produce conditions that favor yeast growth. The
yeast is recycled back into the activated sludge in the
gravity thickener overflow. "Gassing" of the primary clar-
concentrationsin wastewaterinfl uent.
f. Zoogloeas
The presenceofzoogloeal colonies(amorphousor finger-
shaped,Figure 2.5) canbe indicativeof severalconditions.
Both types of zoogloeasare most often observedin high
F/I\4 systems,especially if the waste contains readily bio-
degradable,soluble organic compoundsand the pH is low.
For these reasons,amorphous zoogloea often develop in
activated sludge with selector systems. Zoogloeas are
common in activated sludge treating wastewatercontain-
ing high volatile fatty acid concentrations (e.g., septic
municipal wastewater). In coupled fixed-film/activated
sludgeunits that have no intermediateclarifiers, zoogloeas
often wash into the activated sludge mixed liquor from
the fixed-film reactor. Zoogloeas are not causedby nutri-
ent deficiency.
g. Selector Flocs
Flocs from activatedsludge systemsthat incorporateprop-
erly functioning selectors often have unique features.
Besidesthe presenceof zoogloeas(usually the amorphous
type), these flocs often appearto be tightly structured and
strong. In anoxic and anaerobic selector sludge, the pre-
viously discussed Neisser-positive staining cell clusters
 Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge
i
)-
FfGURE 3.11 Phase contrast micrographs of: (a) yeast, (b) Hyphomicrobium sp., (c) Spirillum sp., and (d) spirochaetes.(Original
magnification 1000x.)
(Figure 3.10c and Figure 3.10d) and individual Neisser-
positive staining cells are seen. Selector sludge is not
always devoid of filamentous organisms,but those present
usually are quite short in length and often have substantial
attachedgrowth - a sign that the fllamentous organisms
are growing slowly. Often their staining reactions are not
typical; all in all, they appear stressed.
h. Nitrifying Bacteria
When nitrifying bacteriaarepresentin significant amounts
in activated sludge, they can be observedmicroscopically
at 1000x magnification as dense, rounded microcolonies
ofbacteria, generally found at floc edges(Figure 2.6). This
was confirmed by Wagner et al. (1998) using fluorescent
in situ hybidization (FISH) methods. The nitrifying bac-
teria were presentin the flocs as sphericalaggregateswith
averagediameters in the range of 6 to 9 pm.
i. DenitrifyingBacteria
Using a phase contrast microscope, it is not possible to
distinguish most denitrifying bacteria from other bacteria
in an activated sludge floc. The exception is the denitri-
fying Hyphomicrobium spp. (Buchanan and Gibbons,
1974) that is identified easily becauseof a unique "bean
on a stalk" morphology (Figure 3.llb). Observation of
this microorganism has proven useful in judging when
ammonia as a nutrient is overdosed in industrial waste-
67
water systems and in preventing the accompanying nitri-
fication-denitrification problems (e.g., floating sludge).
Hyphomicrobium spp.also grow in activated sludge treat-
ing wastewatercontaining methanol and other C, organic
compounds.
j. Spirochaetes, Spirillum, and Flexibacter
Influent wastewater or aeration basin septicity can be
detectedby the presenceof high amountsof Spirillum spp.
(Figure 3. I I c), spirochaetes(Figure 3. 11d), or Flexibacter
spp. (Figure 2.29a) found free in the fluid between acti-
vated sludge flocs. These bacteria grow preferentially at
high organic acid concentrationsand low dissolvedoxygen
conditions associatedwith septicity.
k. ExocellularMaterial
The presence of large amounts of exocellular slime in
activatedsludgecan be detectedby two methods.A micro-
scopic examination of the activated sludge stained with
India ink will reveal large areasofthe floc that are inpen-
etrable to the ink particles (Figure 1.6) in which cells
surrounded by exocellular material can be seen. These
exocellular slimes usually contain polysaccharides and
proteins and their presenceand amounts can be verified
by the Anthrone test for total sludge carbohydrates
(Table 2.12) and the modified Lowry method for proteins
(Table2.13).
6B Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
FICURE3.12 Phasecontrastmicrographsof (a) unicellular Chlorella spp. and (b) filamentots Uronema sp. (Original
magnification1000x.)(Seecolor insertfollowing page70.)
Activated sludge from domestic wastewater treatment
plants normally contains 10 to 20Vo total carbohydrate
(expressedas glucose on a dry weight basis) and usually
abott 50Voprotein on a dry weight basis.Industrial waste-
water treatment activated sludge may contain low amounts
(<l|Vo of dry weight) of polysaccharideif sffessedor con-
taining large amounts of inorganic particulates. High
amountsof carbohydrate(>25V0of dry weight) occur when
the activatedsludge is nutrient limited. In severecasesof
nutrient deficiency, carbohydrate levels up to 90Vo have
beenobserved.Viscousbulking usually occurswhen waste-
waters high in readily metabolizable,soluble organics are
treated under nutrient (N and/or P)-deficient conditions.
Apparently the exocellular polysaccharidesare productsof
a shunt metabolism or unbalanced growth. They appear to
be formed by activated sludge microorganisms (both floc
formers and filaments) when they cannot produce N- and
P-containingcell material due to the lack of nuffients.
Overproduction of polysaccaride has also been
observed in domestic wastewater activated sludge plants
that are not nutrient deficient. The polysaccharideproduc-
tion is associatedwith very high DO uptake rates (often
>100 mg Orlg VSS/h). These high metabolism rates prob-
ably lead to unbalanced growth becausethe bacteria are
unable to obtain nutrients as rapidly as the organic mate-
rials are processed.In severecasesof viscous bulking, the
activated sludge is slimy to the touch and will hang in
viscous "stringers" from the end of a sampling pipette.
Often, the aeration basin or final clarifier will be covered
with a viscous, sticky, solids-rich foam several feet thick.
In one severe case, the slime "stuck" to the secondary
clarifier sludge removal mechanism and would not flow
into the RAS withdrawal line.
In addition to viscous sludges containing large
amounts of exocellular slime, other microscopic signs of
nutrient deficiency include:
. Large clumps of Neisser-positivestaining cells
in the flocs in the absenceof an anaerobic/aero-
bic sequencein the aerationbasin, often present
as tetrads
. Large amounts of intracellular PHB granulesin
the floc bacteria and filaments
. Unusual Neisser staining reactionsof some fila-
mentousorganisms(e.g.,Type 0041, Tlpe 0675,
'and 1L hydrossis often stain lightly Neisser-
positive when nutrient deficient)
. Extensive gonidia and rosette formation by
Thiothrix spp. and Type 021N
l. Algae
The finding of algae (Figure 3.12a and Figure 3.12b) in
activated sludge is rare. These organisms do not grow in
activated sludge becauselight does not penetrate signifi-
cantly into the mixed liquor. Algae in activated sludge
usually originate from another treatment unit in the pro-
cess stream and are fed or recycled into the activated
sludge process. For example, algae can grow on the top
layers ofbiofilters and be flushedthrough the biofllter into
an activated sludge unit in a coupled biofilter-activated
sludge system.
Algae can grow in large amountsin lagoons.If there is
a supematant recycle stream from a sludge lagoon back to
the activated sludge process, algae can enter the activated
sludge system in this way. The washing down of algal
growth on secondaryclarifier weirs can introduce algae into
activated sludge when the wash water is recycled. If the
activated sludge system is followed by secondaryeffluent
filters, algae growing on or removed by the filters can return
to the activated sludge in recycled filter backwash. In one
instance, large amounts of algae in activated sludge resulted
from discharge of water treaffnent plant filter backwash to
the collection system of the wastewater treatment plant.
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge
4. FilamentousOrganisms
a. Relationship to Causes of Bulking
Microscopic examinationof the types,abundance,condi-
tion and growth forms of filamentous organismsprovides
a wealth of information on the nature and causesof solids
separation problems in activated sludge. The ability to
draw conclusionsfrom microscopicexaminationof fila-
mentous organismsis possible becauseof studies that
(1) correlated filamentous organism types observed in
activatedsludgewith wastewatercharacteristics,operating
conditions, and aerationbasin configurations;(2) deter-
mined the growth characteristics and metabolism of
filamentous organismsisolated in pure culture; and (3)
investigatedactivatedsludgefilamentousorganismsin situ
using molecularbiological and physiologicalmethods.
It is rare that an activated sludge sample fiom a plant
with bulking or foaming contains only one filamentous
organismtype. More than one type is usually presentin
significantor dominantquantities(an abundanceof "very
common" or greater)and severalothers may be present
in smaller or secondaryamounts ("common" or less).
Correlation of the causeof bulking or foaming is made
with the dominant filamentousorganismtypes.The find-
ing of severaldominant filament types at one time often
indicates unstableor varying conditions in the aeration
basin.This often happensin activatedsludgefrom indus-
t r ial was t ewa te r tre a tme n t p l a n ts (Ei k e l b oom and
Geurkink, 2002). The filamentous organisms present in
smaller amountsshould not be ignored. Sometimesthey
can provide additional information such as the presence
of a particularwastewatercharacteristicor treatmentprocess
or "left-overs" from a previouscondition. Some filamen-
tous organismsare specific indicatorsof a particular set
of conditions,while othersare presentunder a variety of
conditions.In the latter case,it is important to pay close
attentionto the growth conditions of the filamentousorgan-
isms becausethey may provide cluesabout which of several
possible factors may cause the growth. The factors influ-
encing filamentousorganismgrowth in activatedsludgeare
the same as those influencing the growth of all other types
of microorganisms in activatedsludge:(1) wastewater(sub-
strate)characteristics,(2) processdesignparameters,and
(3) treatmentplant operatingconditions.
Wastewater characteristics that influence filamen-
tous organism growth in activated sludge are:
. Nutrient balance(e.g.,N and P concentrations)
. Readily metabolizablesoluble organics (e.g.,
organic acids, especially the low molecular
weight fatty or volatile acids and soluble simple
sugar carbohydrates)
. Dissolved sulfide
69
. Lipids (especially those containing unsaturated
fatty acids such as oleic acid)
. Particulate substratessuch as starch
Process design parameters important in determining
the levels and types of filamentous organismsin activated
sludgeare:
. Net growth rate (MCRT, SRT, FA4)
. Aeration basin configurationand redox condi-
tions
. Wastewaterf'eedingregime,especiallywhether
continuousor intermittent(e.g., SBRs)
. Foam trapping features
. Upstreambiological treatmentunits, sewersur-
faces,and in-plant surfaces
Plant operating conditions related to filamentous
organismgrowth in activatedsludgeare:
. DO concentration
' pH
. Temperature
b. Nutrient Balance
N and P deficienciescan encouragethe growth of several
filamentousorganismtypes.Type 02l N and Thiothrix spp.
may be encounteredat low N conditions.S. natans,
H. hydrcssis,and N. limicola III may grow in conditions
of P deficiency.When growing under nutrient-deficient
conditions, Type 02lN and Thiothrix spp. ofien exhibit
rosettesand gonidia (Figure 2.19) and usually contain
intracellularPHA granules.
Other signs of nutrient deficiency include the detec-
tion by India ink reversestaining of significant amounts
of exocellularmaterialin the activatedsludgefloc and on
dispersedbacteria.In sludgeswith significantamountsof
India ink-impenetrableexocellularmaterial,the Anthrone
analysis will usually show higher than normal carbohy-
drate levels.
c. Readily Metabolizable Soluble Organics
Most of the filamentousorganismson which pure culture
experimentshave beenconductedcan grow well on fairly
simple, soluble, readily metabolizableorganic substrates
such as low molecularweight fatty acids and simple sug-
ars.This doesnot necessarilymean that thesefilamentous
organismsgrow on thesesubstratesin activatedsludge.
This information can be used to associatesome fila-
mentous organismswith this general type of substrate.
Thus, the filamentous organismsthat appearto be favored
by soluble,readily metabolizablesubstratesareS. natans,
Type 1701, Type 021N, Thiothrix spp., 1L hydrossis,
N.limicolaII, N. limicola III, and Type 1851.S. natans,
70
M anua lo n C a u s e sa n d C o n tro lo f A c ti v atedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Type 1701,N. limicola II, and Type 1851 are often seen
when simple sugars and soluble starches are present in
wastewater.Type 021N, Type 0411, Thiothrix spp.,Type
0914, N. limicola II, and N. limicola III grow on waste-
waters containing volatile fatty acids such as acetic, pro-
pionic and butyric acids. These can be generatedby the
fermentation that takes place in the sewersystemin equal-
ization basins or in primary clarifiers to produce septic
wastewater.
d. Sulfide
Five filamentousorganismscan utilize hydrogen sulfide
as a source of energy (oxidizing it to sulfur and then
depositingthe sulfur as intracellulargranules).Theseare
Thiothrix spp., Type O2lN, Beggiatoa spp., and Type
0914. When growing on sulfide, theseorganismsexhibit
intracellularsulfur granulesin situ or in the S test. Some
of these organisms(Thiothrix spp. and Type 021N) use
both sulfide and the low molecular weight organic acids
produced in septic sewage and, thus, their growth is
strongly encouragedby septicity (Richard et a1., 1983,
1985). These organisms appear to be unable to utilize
poorly soluble sulfidessuch as iron sulfide.
Beggiatoa spp. are found in fixed film reactors and
can enter activatedsludge by washing through from the
first stageof a coupledfixed-film activatedsludgeprocess.
Beggiatoa spp. can be seen in the heavy white tuft-like
growthsseenin the first stagesof oxygen-deficient(organ-
ically overloaded)RBC plants. The white color of the
growth is due to the intracellular sulfur granules in the
Beggiatoaspp. Scrapingthe white growth below the sur-
face will reveal a black septic layer. Thus Beggiatoo grows
on the aerobicsurfaceof a septicbiofilm, obtainingsulfide
from below and dissolved oxygen from the solution to
oxidize the sulfide to sulfur.
e. Lipids
Recent work (Nielsen et al., 2002; Tandoi et al., 1998;
Mamais et al.. 1998:Andreasenet al., 1998)indicatesthat
M. parvicella has the ability to take up and store oleic
acid, a long chain fatty acid (and to someextent palmitic
acid), under anaerobicconditions.Lipids containingoleic
acid can be hydrolyzed by M. parvicella using a cell
surface-associatedlipase enzyme. Anecdotal evidence
indicates that growth of nocardioforms in activatedsludge
is associatedwith the presenceof lipid materialsin the
wastewaterfeed (Chapter5).
f. Other Particulate Substrates
Type 0041, Type 0675, and Type 0092 filamentousorgan-
isms apparently can grow on slowly biodegradeable(or
perhapsevenparticulate)substrates. Theseorganisms(and
M. parvicella) were the most common in South African
BNR-activated sludge plants with initial anaerobic and/or
anoxic zones (Blackbeard et al., 1986b). Ekama and
Marais (1986) postulatedthat slowly metabolizableand
P roblem s
particulate organic matter degradationrates are very low
in anaerobicand anoxic zonescomparedto the ratesunder
aerobic conditions.Becauseof this, the particulatesand
slowly metabolizable substratesare transportedto the aer-
obic zones of the BNR plants where they are hydrolyzed
and produce low concentrations of soluble organics that
favor the growth of these filamentous organisms.
Richard(1986) studiedthe fatesof filamentousorgan-
isms in aerobic digestersat ten domestic wastewateracti-
vated sludgeplants in Colorado.While most filamentous
organismspresentin the mixed liquor rapidly degraded
and disappearedin the aerobicdigesters,Type 0041, Type
0675, Type 0092, M. parvicella, and nocardioformsper-
sisted.M. panicella and the nocardioforms grew to prob-
lem levels and causedfoaming in some aerobicdigesters.
The hydrolysis of particulate materials to produce low
levels of soluble organics may be responsiblefor the
growth of thesefilamentsin aerobicdigesters.
Eikelboom et al. (1998) found that Type 0041 and
Type 0675 were favored in BNR-activatedsludge plants
in which raw wastewater and RAS were mixed prior to
enteringthe aerationbasin. On the basis of this observa-
tion, theseworkers suggestthat theseorganismsgrow on
particulate substrates.
g. Case Study
An example of how the nature of a wastewatersubstrate
affects filamentous organism populations in activated
sludgeis providedby the work of Richard (1997) on pulp
and paper-activatedsludge systems.From 1982 through
1990,29pulp and paper-activated sludgeplantswith bulk-
ing sludgewere examined;resultsare shown in Table 3.3.
A similar surveyof 80 plantsin 1996producedthe results
shown in Table 3.4. The most striking difference in the
resultsofthese surveysis the largedecreasein occurrence
of Type 0675 and the large increasein the occurrence of
Thiothrix spp., N. limicola II and III, and Type 0914 in
1996.
Based on these filamentous organism analyses,the
causefor bulking in theseplantswas diagnosedas shown
in Table 3.5. A large increasein the incidenceof septic
wastewaterwith the presenceof sulfide and readily metab-
olizable, low molecular weight organic acids occurred.
Low F/M filamentousorganisms,especiallyType 0675,
decreasedsignificantlyover this time period.The reasons
for these changes can be traced to changes both in the
pulping processand in the raw materials used. The major
pulping processchangewas the reduction or elimination
of chlorine for pulp bleaching (to prevent the formation
of chlorinated organics) and its replacementby oxidants
such as hydrogen peroxide. These bleaching agentsintro-
duce more biodegradablesoluble organics into the waste-
water, leading to a more rapid exertion of BOD in the
plant sewers and primary treatment units, and increasing
the propensity of the wastewater to turn septic prior to
Color Figure1.6
Phasecontrastmicrographs of reversestainingof activatedsludge:(a) viscousflocsin anunstained (b) normalflocsstained
preparation;
with Indiaink; (c) and(d) viscousflocsstainedwith Indiaink. (Originalmagnifications 100x, (a),(b),and(c); 1000x,(d).)
ColorFigure2.4
particlesin floc: (a) phasecontrast,
Microgriphsof nonbiological (Originalmagnification
(b) directillumination. 100x.)

Color Figure2.5
200X, (a);
Phasecontrastmicrographsof zoogloeas:(a) and(b) fingered,(c) and(d) amorphous(globular).(Originalmagnifications
1000x,(b); 1@x, (c);and1000x,(d).)
 ColorFigure2.15
Phase micrographs
contrast of intracellular sp.,(c)Tlpe 02lN, and(d)Type0914.
(a)Thiothrixsp.,(b)Beggiatoa
sulfurgranules:
(Originalmagnification
1000x.)

Color Figure2.16
Direct illumination micrographof PHA staining. PHA granulesare dark blue/black.(Original magnification 1000x.)

I
Color Figure2.17
Direct illumination micrographs of Gram staining of filamentous organisms: (a) improperly decolorized floc retaining Crystal Violet,
(b) Gram-negativeNostocoida limicola II, (c) Gram-variableType 0041, (d) Gram-variableType 1851, (e) Gram-positivenocardioform,
and (f) Gram-positive Microthrix parvicella organism. (Original magnification 1000x.)
 tr ru4; .tF#
+.5

4tr,

,H
'uqu
I
itl

Wllri ,

."#'
p

Colo r Fig ure2 .18

Direct illuminationmicrographsof Neisserstainingof filamentousorganisms.(a) Neisser-negative Type 021N, (b) and (c) Neisser-
negativewith positive granules(nocardiofbrmorganismsandMicrothrir parvicellu, respectively),(d) and (e) Neisser-positive(Type
0092 and Nostoutida linticrila II, respectively),(f) talse Neisser-positivestaincovering filament,Type 0041. (Original magnitication
I 00 0x. )
 C ol orFi gure2.21
Phasecontrastmicrographs<:f: ('ttlSlthucnttiltt,\n(tuns. (b)
(Original
Typc 170|. (c) Huli,scotrtotobutttr/rlr/rur.r,rl,r.
r r a g n i l i c a t i o nI ( X X ) x ).

tr tr

Color Figure2.22
Phasecontrast of: (a)Type021N.(b) Type02lN with sulfurgranules(c) Beggiatoasp. (d) Be,qgiaroasp. with sulflr
rnicrographs
(Original
granules. magnification1000x.)
ColorFigure2.23
Phaseconfrast
micrographs (c) ThiothrixII, and(d) ThiothrixII with sulfur
of: (a)Thiothrixl,(b) ThiothrkI with sulturgranules,
(Originalmagnification
granules. 1000x.)
 H, tr
"-i
*{
,4{
"c
*--+ r
.t
"
i .{..}' -4-f
' >-
l'rrrt
"ttL'.J.|t'

Color Figure2.24
P has cc ont r asrn (Ori gi nalmagni fi cati on
t i c ro g ra p hosl ' : (a ) T y p e0 9l 4 and (b) Type09l 4 w i th sul furgranul es. 1000X. )

[.i.

,".'-h,q.f l

*'s.i{fu.*,*,',
il
lr , '

tl r

Color Figure2.28
Phase
contrast of: (a)Microrltrixparvicella,(b) Nocardioforms,
micrographs (c)TypeI 863,and(d)Type0211. (Originalmagnification
1000x . )
 E I {,

l,

-*

Color Figure2.29
Phasccontrast rnicrographs /('r sp.,(b) lJacrl1rr,r
of: (a) Flc.ribtl( and (d) fungus.(Original
sp. (c) 0:;cillutoriasp. (cyanophyceac),
I ( X X)
m agnif ic at ion X .)

Color Figure 2.30

14 ,iirl identiflcationof nitrifying bacteriain activatedsludge:(a) simultaneous in .tilu hybridizationwith Cy3-labelledprobeNmV
and FLUOS-labelledprobe NEU. Nl/ro.vrcoc't:tr,s mobilis cells appearyellow; (b) simultaneousin si/u identificationof Nitrosot'ttcctrs
mobili.s anclNitntsportt-likebacteria 'afterin situ hybridization with FlUOS-labelled probe NmV (green)and Cy3-labelledprobe S-
*-Ntspa-1026-a-A-18 (red).(From Juretschko. S. et al. (1998),Appl. Environ.Microbiol.,64,3042.With permission.)
Color Figure2.31
Phasecontrastmicrographsof commonflagellatesandamoebae foundin activatedsludge:(a) Monassp.,(b) Bodo sp.,(c) Polychaos
sp.,and(d) Arcella sp. (Originalmagnifications1000x, (a) and(b);400x, (c) and(d).)
 Color Figure2.32
Phasecontrastmicrographsof free andstalkedciliates:.(a) Aspidiscasp.,(b) Parameciumsp.,(c) Tokophyrasp.,and(d) Podophyra
sp.(Originalmagnifications
400x, (a); 200x, (b), (c), and(d).)

t_
Color Figure2.33
Phasecontrastmicrographsofstalkedciliates:(a) Operculariasp.,(b) Vaginicola sp., (c) Vorticella sp., and (d) Epistylis sp. (Original
100x, (a);200x, (b),(c),and(d).)
magnifications

Color Figure2.34
Phasecontrastmicrographsof rotifers. (Originalmagnification200x.)

l
 ColorFigure2.35
Phasecontrastmicrographs (a) bristleworm(Aeleosoma
of invertebrates: sp.),(b) tardigrade
(waterbear,Macrobiotussp.),
(c)gasterotrich
(Chaetonotus
sp.),and(d)hydrachnidnematode.(Originalmagnifications
100X,(a);200X,(b)and(d);400x,(c).)

t-

ft .

I
Color Figure3.7
Phasecontrastmicrographsof nonmicrobialparticles:(a) amorphousorganicmatter,(b) iron sulfide,(c) fenic iron precipitate,
200x, (a) and(b); 1000x(c); 100x, (d).)
and(d) activatedcarbon.(Originalmagnifications
Color Figure 3.'l 0
Neisser-positivecell clumps in activatedsludgeflocs: (a) tetrad,phasecontrastof wet mount; (b) tetrad,direct illumination Neisser
stain; (c) phosphate-accumulatingorganisms(PAOs),direct illumination Neisserstain; and (d) G bacteria,direct illuminarion Neisser
stain.(Original magnification1000X.)

Color Figure3.12
of (a) unicellularChlorellaspp.and (b) filamentousUronemasp. (Originalmagnification1000X.)
Phasecontrastmicrographs
ColorFigure5.1
Phasecontrastmicrographs andM. pamicell.afoansi (a)and(b) nocardioforms
of nocardioform (possiblyG. amarae),(c) and(d)
S.pinensis, (e)
and and (f) M. (Original
pamicella. magnifications
100x, (a),(c),and(e); (b),
1000x, (d),and(f).)
 Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge 71

TABTE3.3 TABTE3.5
Occurrenceand Rankof FilamentTypesCausing Comparisonof Causesof Filamentous
Bulking in Pulp and PaperWastewaterActivated Bulkingin Pulpand PaperWastewater
SludgeSystems, 1982 through1990 ActivatedSludgeSystems,1982-1990
Rank FilamentType Number of Plants Percenl
and 1996
I Type0675 16 55 Percentof Plants
2 Type1701 8 28
Cause of Bulking 1982-1990 1996
2 Type1851 8 28
3 ThiothrixIl 7 24 Septicity 21 68
3 Type0041 7 24 Low DO 22 9
4 N. limicola II 5 17 Low F/M 49 l0
4 Type021N 5 17 Low N/P 8 13
4 H. hydrossis 5 17
5 S. natans 2 7 Source: From Richard, M.C. (1997), Proc. 1997

5 ThiothrixI 2 TAPPI Environ. Conf., p.553. With permission.

7
5 Tlpe 0803 2 7
6 N. limicola lll I 3 plant sewers and primary treatment units to form low
Source: From Richard, M.G. (1997), Proc. 1997 TAPPI Environ. Conf., molecular weight organic acids. These low molecular

L- p. 553. With permission, weight organic acids are ideal substrates for Thiothrix
spp.,N. limicola II and III, and Type 0914.
h. Net crowth Rate (MCRT, F/M)
Table 3.6 shows the approximaterange of MCRT (and
TABTE3.4 F/M) over which various filamentous organisms are
Occurrenceand Rankof FilamentTypesCausing observed.The data were compiled from Richard (1989)
Bulking in Pulp and PaperWastewaterActivated for domesticwastewatertreatmentplants in Coloradoand
SludgeSystems, 1996 Eikelboom (2000) for domestic wastewater treatment
plants in The Netherlands.
Rank FilamentType Number of Plants Percent
Many filamentousorganismsoccur over a fairly broad
range of MCRT. These include Type 1701, S. natans,
I ThiothrixII 44 ))
H. hydrossis,Thiothrix spp., Type 021N, nocardioforms,
z ThiothrixI 36 45
3 N. limicola II 20 25
and N. limicola II. A second group of filamentous organ-
4 Type0914 l9 )4 isms appearonly at high MCRT (low F/M) levels.These
5 H. hydrossis l8 ZJ include M. parvicella, Type 0041, Type 0675, Type 0092,
6 N. limicola III l0 IJ Type 1851,Type 0914, Type 0803, and Type 0581. Two
7 Type1851 8 t0 filamentousorganisms(Type 0411 and Type 1863) seem
a1 Type1701 6 8 to grow well at MCRT <2 d (high F/M). Type 041 I appears
9 Type02lN 6 to be favored by high wastewaterconcentrationsof low
l0 Type0092 4 5 molecular weight organic acids while Type 1863 appears
10 Type041I 4 5 to grow on hydrophobicsubstrates(oil and grease).
10 Type0675 A
5
l1 S, natans 2 3 i. Aeration Basin Configuration and Redox
1t Type0041 2 3 Conditions
1l Type0581 2 3 The growth of many filamentousorganisms(Type 02lN,
12 Type0803 I I
Thiothrix spp., S. natans, N. limicola II, Type 1701,
l2 M. panicella I l
H. hydrossis,Type 1851,and nocardioforms)in activated
t2 Type021I 1 I
l2 Actinomycetes I I
sludge is generally encouragedby the use of uniformly
aerated, completely mixed, continuously fed aeration
Source: From Richard, M.G. (1997), Proc. 1997 TAPPI Environ. Conf., basins.The growth of theseorganismsis suppressed when
p. 553. With permission.
the aeration basin includes an initial high F/M feed zone
(selector),and especially(1) when the selectorand aera-
entering the aeration basin. The raw material change is tion basin are compartmentalized and (2) when the selec-
the increasedand widespreadreuse ofrecycled fiber. This tor is anoxicor anaerobic.
material contains starch, which is readily biodegradable These filamentous organisms are also suppressed
and decomposes anaerobically (by fermentation) in the when an SBR with an unaeratedfeed period and an initial
72 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

TABLE3.6
Relationshipof SpecificFilamentousOrganismsto MCRTand F/M in ActivatedSludge
MCRT,d 1. 9 2. 2 2.5 3.0 4.0 -s.0 8.0 20 50
FA{" 0. 8 0; 7 0.6 0.5 0.4 0.3 0.2 0.1 0.05

Typel70l
S. natans
H. hydrossis
Thiothrixspp.
Type02lN
Nocardioforms
Type0411
N. limicola ll
Type 1863
Type0041
Type0675
M. pamicella
Twna OOO?

Type I 85I
Type0914
Type0803
Tlpe 058I

"F/\4 as kg BOD./kg MLSS, d.

Sources: From Richard, M.G. ( 1989),Activated Sludge Microbiology, Waer Polution Control Federation,Alexandria
VA and Eikelboom, D.H. (2O00), Process Control of Activated Sludge Plants by Microscopic Investigation, IWA
P ub lish in eL. o n d o n .

unaeratedreact period is used.They are not suppressedin
SBRs in which the wastewateris fed over an extended
period and the feed and initial react stages are aerated.
Thesemodesof SBR operationare prone to the develop-
ment of filamentousactivatedsludgejust like continuous
flow, completely mixed systems.
A certain group of high MCRT (low FAvI) filamentous
organisms grow in aeration basins with initial unaerated
(anoxic or anaerobic)zones.These include M. parvicella,
Type 0092, Type 0041, and Type 0675. This meansthat
anoxic and anaerobic selectors may not be effective for
controlling thesefilamentousorganisms.Lind and Lemmer
(1998)reporteda significantshift in filamentousorganism
population to thesetypes in German wastewatertreatment
plants from 1989 to 1996. The shift coincided with the
widespread introduction of activated sludge systems
designed to perform nitrification/denitrification and
enhancedbiological P removal.
j. Wastewater Feeding Regime
Wastewater feeding regimes can be classified as either
continuous or intermittent (discontinuous). Continuous
feeding regimes exist in continuous flow activated sludge
plants; intermittent feeding regimes exist in SBR activated
sludge systems.A discontinuous feeding regime provides
similar biolosical conditions and effects as a selector in a
continuous flow plant. In both systems, the activated
sludge organisms are intermittently exposed to feed con-
ditions. In a continuousflow plant, exposureis through
distanceor locationin the aerationbasin;in an SBR plant,
it occurs over time.
As previously noted, Type 021N, Thiothrix spp.,
S. natans, N. limicola I, N. limicolc II, Tlpe 1701,
H. hydrossis,and Type l85l can grow in continuouslyfed
activated sludge, but not in intermittently fed units (espe-
cially when the feed zone or feeding period is unaerated).
Intermittent feeding does not control the growth of
M panicella,Type 0092, Type 0041, and Type 0675.
k. Foam Trapping Features
This topic will be dealt with in greater detail in Chapter
5. Surface trapping, caused by features such as surface-
intercepting baffles or walls and subsurfaceliquid with-
drawal will favor the retention and increasethe population
of filamentous organisms that float, e.g., nocardioforms
and M. parvicella. Conversely, systems with free water
surfaces will disfavor the development of large popula-
tions of filamentous organisms that float. The effects of
foam trapping often are exacerbatedby foam removal and
recycle to a point upstreamofthe activatedsludgeaeration
basin. These actions reseedthe activated sludge with the
foam-producing filamentous organisms and leading to an
even greater foaming problem.
Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge
l. Upstream Biological Treatment Units, Sewer
Surfaces, and In-Plant Surfaces
When a biological treatment unit is sited upstream of an
activatedsludgeplant, filamentousorganismsthat originated
in the upstreambiological treatmentmay be presentin the
activatedsludge.Common examplesin activatedsludgeare
fungi and Beggiatoa spp. from an upsffeam biofilter in a
coupled biofilter-activatedsludgeplant. This type of "seed-
ing" is especially noticeableif no intermediateclarifier is
situatedbetweenthe biofilter and the activatedsludge.Thus,
the mixed liquor in a TF/SC processoften contains many
filamentousorganismsthat originate in the upstreambiofil-
ter. RBCs upstreamof an activatedsludge systemwill also
contribute such organisms,especiallyBeggiatoa spp.
The same type of seedingand occurrenceof filamen-
tous organismscan appearin two-stageactivatedsludge
processesin which organisms from the first stage unit
appearin the mixed liquor of the secondstagesystem.It
is not necessary for the upstream biological treatment
processto be at the samelocation as the activatedsludge
plant. Thus, filaments grown in biological pretreatment
units at industrialfacilitiesthat dischargeto a sewersystem
can reachan activatedsludgeplant treating this wastewater.
Kappeller and Gujer (1994) suggestedthat filamentous
organisms such as S. natans growing on sewer walls can
seedactivatedsludge systemstreating the wastewater.The
seeding of activated sludge by recycling trapped foam-
containingnocardioformsor M. parvicella was mentioned
earlier and will be discussedin detail in Chapter5.
Seeding of activated sludge units with filamentous
organismscan be a seriousproblem in laboratoryand pilot
plant units becausethe surface area-to-volumeratios in
the pilot plant unit, feed lines, and storagevesselsare
much higher than for full-scale activatedsludge systems.
Unless specialcare is taken, filamentousorganismssuch
as S. natans,Type 1701, and Thiothrix spp. will grow on
the surfacesof the feed storage vessel and the feed lines
and will seedthe activatedsludge and causebulking. This
was first noticed by Gabb et al. (1989) when S. natans
caused bulking in laboratory-scalenitrifying and BNR
activatedsludgereactors,whereasthis organismwas never
observed in full-scale systems operated under the same
conditions and on the samewastewaters.
A surveyofthe literatureon pilot scaleactivatedsludge
systems utilized for bulking control studies revealed the
widespreadoccunence of low DO filamentous organisms
under a wide range of operating conditions (Table 3.7).
Seedingfrom growth in the feed system is suspected.The
appearanceofthis artifact can lead to erroneousconclusions
on the effectivenessof bulking control measures.Thus, if
activatedsludge settlespoorly becauseof S. natans growth
(an experimental artifact) and measures are taken that
improve the sludge settling, one will be led to the conclu-
sion that these measureswill provide an effective bulking
73
control in a full-scale systemoperatedunder the samecon-
ditions as the laboratory unit. This may not be the case
becauseall that the laboratory experiment achieved was
controlling an artifact of the experimentalsystem.
These seeding problems can be prevented by regular
cleaning of feed storagevessels,including scrubbingthe
walls with a sodium hypochlorite solution and regular
cleaning of feed lines by immersion in or flushing with
sodium hypochlorite solution. It is good practice to have
a duplicate set of feed lines with one set in service and
the other set cleanedand ready to be placed in service.
The activatedsludge in these systemsshould be micro-
scopically observed daily for the types of filamentous
organisms that grow on surfaces.
m. DO Concentration
Type 1701, S. natans, H. hydrossis, and M. parvicella are
associatedwith low DO. Type 1701, S. natans, and
H. hydrossis are found at low to moderate MCRTs (2 to
l0 d) while M. pamicella appearsat high MCRT (higher
than approximately12 d). It will be seenin Chapter4 that
low DO is a relative term becausethe DO concentration
associatedwith low DO filamentousorganismgrowth is
a function of the F/M of the activatedsludse.
n. pH
The presenceof large numbers of fungi in an activated
sludgeindicateslow pH (<6.0) and suggeststhat the influ-
ent containsstrongacid, indicatingan industrialdischarge
of acid. Some nitrifying activatedsludgesystemstreating
low alkalinity wastewatercan exhibit low pH, as can oxy-
gen-activatedsludge systems treating low alkalinity
wastewater.Fungalbulking hasneverbeenobservedunder
these conditions. As previously noted, fungi can be
observedincidentallyin the mixed liquor of coupledfixed-
film activatedsludge systems.Fungal bulking has been
observedwhen fungi occur in high numbersin wastewater
from an upstreamindustrial pretreatmentprocess.
o. Temperature
Within the normal ambient temperature range found in
activated sludge aeration basins (8 to 25'C), it can be
generally stated that filamentous organisms grow more
rapidly as the temperatureincreaseswithin this range. One
exceptionis M. parvicellawhich in many instancesseems
to predominate in high MCRT-activated sludge at low
temperatures(10 to 15"C). A common observation in
colder climates is that foaming in the winter is caused
largely by M. parvicella but in the summer the major
filamentous organisms causing foam are nocardioforms.
Eikelboom (2000) suggests that M. parvicella and
Type 0092 also show a similar switch in dominance in
very high MCRT oxidation ditches. M. parvicella is
dominant at temperaturesbelow 15'C and Type 0092 is
 74 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
P r oblem s

TABLE3.7
ConditionsunderWhich S. nafansWasa Dominant FilamentousOrganismin LaboratoryScaleActivatedSludge
Units
Aerator FlM,g COD/g FlM,gBODr/g Aerator
Author BOD'/N COD/N DO, mg OrlL BOD'/P COD/P MCRT,d HRI h vss,d vss,d MLSS,g/t

Clesceri (1963) 8 0.6.

Hattingh (1963) >22 > 168 23
Chudobaet al. 8 l.t
(1913a)
Houtmeyerset al. 6 'l 1 4
(1980)
P a l me t a l . ( 1 9 8 0 ; d 0.1-5.5 1.9-11 0.38-1.72
Verachtert et al. I 0.4
( 1980)
Ch i e s aa n d l r v i n e 8 0.56"
( 1e85) -
Le e e t a l . i 1 9 8 2 ; 6 .8 16.3 4.5-16" 15 19 0.31-1.00 0.134.42 3-3.5
Rensink et al. 10 58 r5.9 92.7 0.025-2.00f 2
( l 982)
van den Eynde et - 83 I
al. (1982a)
Wu et al. (1983) 20 and 69 95 and28 7-8 24 0.2-0.4 0.1-0.3 l-3
G a b be t a l . ( 1 9 8 5 ) 5-8 t2-14 2.5 0.1 z-3
van Niekerk 1 5 .1a n d 2 2 .9 6l and 89 3.54.5 20_30 0.2-0.3
( l 985)
Burkeet al. t2 .6 24 24 3.8 1.26 -
( l 986).
Still et al. (1986)" <^ t-'7 20 24 0.35 11

Srill et al. i1986)h 5.4 2-3 20 16 0.31 -

' Completely aerobicconditions.
h 4 h cycle with I h anoxic, 3 h aerobic.

' High rate Phoredox. When onethird of the feed was bypassedto the aerobic reactor, abundant S. natans occurredi with all feed to the anaerobic zone
S. natans abundancedeclined sharply.
d S. natans grew at high DO with high F/l{ (low MCRT), or at low Flvl (high MCRT) when the DO was at the lower end of the range.

" F/M is g COD/ g MLSS, d.

I F/M is g BOD./ g MLSS, d

Source: From Gabb, D.M.D. et al. (1989), Water Sci. Technol.,2l:1,29. With permission.

dominant at temperaturesabove 15'C. Recent experience p. Summary

with high temperature growth of filamentous organisms
in activated sludge treating pulp and paper and pharma-
ceutical wastewaterssuggeststhat the growth rate of fila-
mentous organismsincreasesup to temperaturesof 35oC.
Between 35 and 40'C, the filamentous organismsseemto
become progressively stressed,frequently growing in a
dispersedstate (not attachedto the flocs). Since most
filamentous organismsare mesophilic, their growth ceases
at about 40'C. However,Type 0914 may be thermotolerant
since Norris et al. (2000) observedits growth in activated
sludge at temperaturesof 50oC.
Table 3.8 is a summary of the causesassociatedwith the
growth of filamentous organismsin activated sludge.This
summary should be read in conjunction with the detailed
discussion of the associationsof various filamentous
organismswith activatedsludge design features,operating
mode, and wastewater characteristicsdiscussedin detail
earlier in this chapter. These causeswill be further elab-
orated in Chapter 4. The sources of the foam-causing
filamentous organisms(nocardioforms and M. parvicella)
will be discussedin Chaoter5.
 Applicationsand Resultsof MicroscopicExaminationof ActivatedSludge /J

TABLE3.8
Summary Associated
of Conditions with Filamentous
OrganismGrowth in ActivatedSludge
Cause FilamentousOrganism
Low DO concentration S. natans
Tlpe 1701
H. lrydrossis

Low F/l\,I $pe 0041

Type0675
'Ilpe 1851
$pe 0803

Elevated low molecular weight organic acid I}pe 021N

concentration ThiothrixI andll
N. limicola I, II and III
Tlpe 0914
Dpe 04ll
Tlpe 0961
Tlpe 0581
Tlpe 0092

Hydrogensulfide Thiothrk I andll

Type02lN
Tlpe 0914
Beggiatoaspp.

Nutrient deficiency Type021N

Nitrogen Thiothrixl andll
Phosphorus N. limicola llI
H. hydrossis
S. natans

Low pH Fungi

'_
i'.
K
t\-
II
 4' Controlof ActivatedSludg"Bulking
and Other SettlingProblems
The countryneeds,and unlessI mistakeits temper,the
country demandsbold, persistentexperimentation. It is
commonsenseto takea methodandtry it. If it fails,admit
it frankly and try another.But aboveall, try something.
Frankin Delano Roosevelt,1892-1945
Addressat Oglethorpe University,Atlanta,
May 22, 1932
I. INTRODUCTION
Since the first edition of this manual was published(Jen-
kins et al.. 1984)much hasbeenlearnedabout factorsthat
affect activated sludge settling characteristicsand how to
control them. It has long been understoodthat activated
sludgesettleabilityis controlledby the physical,chemical,
and biologicalenvironmentof the activatedsludgesystem.
The past three decadeshave seensignificantadvancesin
our understandingof the specificfactorsthat control acti-
vated sludge settleability and the measuresthat plant
designersand operatorscan take to achievebettercontrol
of sludge settleability.This understandingis useful for
resolving activatedsludge settling problems in existing
systemsand for designingnew or upgradedplants.
This chaptersummarizesour knowledgeof the factors
that control activatedsludge settleabilityand how these
factors can be manipulatedto improve activatedsludge
processperformanceand capacity.
II. GENERAL
APPROACH
The presenceof excessivequantitiesof filamentousorgan-
isms is still one of the most common activated sludge
operatingproblems.Their presenceproducesan activated
sludgethat settlesslowly and compactspoorly in second-
ary clarifiersand can reducesystemcapacity.Other types
of settling problemsalso occur and can be diagnosedand
dealt with using the proceduresoutlined below. It almost
goeswithout sayingthat the best approachfor controlling
activatedsludgesettlingand thickeningcharacteristics and
ensuringlong-term,problem-freeplant operationis using
appropriate design and operating methods.
However,becausemany of thesetechniqueshave been
developed relatively recently, even well designed and
operatedplantscan experienceproblemsfrom unusualor
unanticipatedoperating conditions. Consequently,many
existing activatedsludgeplants are faced with solids sep-
aration problems. The following general approach is rec-
ommendedto correct theseproblems:
. Performthe microscopicexaminationprocedure
outlined in Chapter 2 to determine the nature of
the activated sludge floc and the amounts and
identitiesof filamentousorganism(s).
. Use the results of the microscopic identifica-
tion, the information in this manual, and a
knowledge of plant operating conditions and
wastewater characteristics to determine the
probable cause(s)of the poor sludge settling
and/orthickeningpropenies.
. If the probablecausecan be rectified without
major changes,make the required operating
changesto addressthe problem.For example,if
septic wastewateris indicated,wastewaterpre-
chlorination may be initiated. If the probable
cause is nutrient deficiency,determine which
nutrients are deficient by analysesof influent,
effluent,and activatedsludgesolids.Rectify the
deficiencyby increasingthe feedrateof existing
nutrient supply systemsor installing nutrient
additionfacilities.If elevatedeffluentsuspended
solids are causedby poor flocculation in the
aerationbasin, it may be possible to improve
flocculationby changingaerationpatterns.
. Plants with sludge bulking and other settling
problernsmay require major design or operat-
ing changesthat could take a long time to
implement (e.g., installing additional aeration
capacity,changingaerationbasinconfiguration,
industrial wastewater control, etc.). After
changes are made to discouragefilamentous
organism growth, sludge settleability may
improve only slowly. Afier a changein operat-
ing conditions to retard filamentousorganism
growth, the activated sludge microbial popula-
tion may change at a rate proportional to the
culture washout rate (or mean cell residence
time. MCRT). In a completelymixed system,a
period of approximately 2 to 3 MCRTs are
required for a bulking activatedsludge to return
to a nonbulking condition (Palm et al., 1980).
77
7B M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Data on the effects of selectors suggest that
more than 2 to 3 MCRTs may be required for
them to exert their full effect on sludge settle-
ability (Wheeleret a1.,1984;Boe et al., 1996).
. Rapid, nonspecificmethodsmay be neededto
eliminate sludge settling and bulking symp-
toms. Thesemethodsfall into three categories:
(i) Manipulationof RAS flow ratesand waste-
water feed points to the aerationbasin
(ii) Addition of chemicals or inert solids to
enhancethe activatedsludgesettling rate
(iii) Addition of disinfectantsto the activated
sludge to selectivelykill the filamentous
organismsthat causebulking
None of these methods change the conditions that
causedthe poor settleability,and settling propertieswill
most likely deteriorateagain when these measuresare
discontinued.
B UTKI NG
III. RAPID,NONS P E CIFIC
CONTROLMETHODS
oF RASFt-owRnrrs
A. MnxrpumnoN
AND AERATIoNBASINFrro POITTS
The adverseeffectsof bulking sludgeoften can be mini-
mized by proper managementof the activatedsludgesys-
tem, particularly if it is under-loadedand/or the ability to
utilize certain processoptions is built into the design.To
understandhow these processmanagementtools can be
used to deal with bulking sludge it is necessaryto:
. Figure 4.1 illustrates the activatedsludge process.The
Review activated sludge secondary clarifier
operatingprinciples,particularly as they relate
to the secondaryclarifier thickening function
. at flow rate Q together with RAS at flow rate Q,. The
Present techniques for determining the sus-
pendedsolids (SS) handling capacityof a sec-
ondary clarifier, and how it is influenced by
sludge settling characteristics
. Review secondary clarifier and aeration basin
operating techniquesfor maximizing secondary
clarifiercapacityin systemswith bulking sludge
1. SecondaryClarifier Operating Principles
The two basic functions of activated sludge secondary
clarifiers are clarification and thickening. Clarification is
the removal of activated sludge flocs to produce a clear
overflow that meetsdischargestandardsand doesnot over-
load downstreamtreatmentprocesses(e.g.,tertiaryfilters).
A clarification failure results in elevatedeffiuent SS con-
centrationsthat may exceedeffluent dischargerequirements
even though a low sludgeblanket is maintained.Clarifi-
Pr oblem s
cation failure can be causedby poor bioflocculation,floc-
break up in the aeration basin and other locations prior to
settling in the clarifier, or hydraulic deficiencies in the
clarifier. The factors influencing clarification failure are
discussedlater in this chapter.
Thickening of the settled activated sludge SS is
required so that the solids can be returned in the RAS to
the aerationbasin. Ifthe activatedsludge SS are not thick-
ened and then removed from the secondaryclarifier at a
rate at least equal to the rate at which they were added,
they will accumulatein the secondaryclarifier. Eventually
the clarifier will fill up with SS and the SS will spill over
into the effluent. Since bulking affects the ability of the
activatedsludge SS to thicken, the thickening function and
thickening capacity of the secondary clarifier are more
affectedby sludgebulking. When thickening characteristics
deteriorate,the mass of SS retained in the system (sludge
or SS inventory)candecreasedueto SS lossin the effluent.
Even without such losses,however,poor thickening
characteristicscan reduce aeration basin SS inventory
becausethe SS accumulatesin the secondaryclarifier
insteadof returningto the aerationbasin.In severecases,
the reducedaerationbasinSS inventorycan resultin lower
pollutant removal capacity and a further deteriorationin
settling characteristicsbecause of the increased F/M.
Improvementof activatedsludgethickeningpropertiesby
control of excessive filamentous organism levels can
restoreplant capacity.In some instances,plant capacity
has exceededdesign values after improvementof sludge
thickening properties.
2. Activated Sludge ProcessSchematic
and Definitions
aerationbasin has a volume V and the secondaryclarifier
has a surface areaA. Wastewaterenters the aeration basin
aeration basin SS concentration(MLSS) is X. and the
RAS, SS is X.. Activated sludge SS are applied to the
secondaryclarifier at a flow rate of (Q + Q.).
It is assumedherethat sludgeis wasted(wasteactivated
sludge,WAS) from the RAS at a flow rate Q* and that the
WAS has the same SS concentration as the RAS (X.).
Effluent is dischargedfrom the secondaryclarifier at a flow
rate of (Q - Q*) and contains an SS concentrationof X".
3. SecondaryClarifier Process
Operating Relationships
In developingrelationshipsto describethe secondaryclari-
fier thickening function, certain simplifying assumptions
can be made. First the WAS flow rate Q* usually is small
comparedto the influent flow rate Q, and its effect on effiu-
ent flow can be isnored. Second.the ouantitiesof activated
Control of ActivatedSludgeBulkingand Other SettlingProblems 79

Secondary
Aeration basin clarifier

Return activated sludge Wasteactivated sludge

(RAS) (wAS)

DEFINITIONS

A = Clarifier surface atea

V = Aeration basin volume
Q = Influent flowrate
Q. = RAS flowrate
Q* = WAS flowrate
X = Aeration basin MLSS concentration
X. = RAS suspendedsolids concentration
X- = Effluent suspendedsolids concentration

FIGURE4.1 Schematic sludgeprocess.

diagramof the activated

sludgeSS dischargedin WAS and in the secondaryeffluent
are small comparedto the quantity of activatedsludge SS
applied to and withdrawn from the secondary clarifier.
Using theseassumptionsand assumingthat SS do not
accumulatein the secondaryclarifier, a simplified SS mass
balance around the secondarvclarifier at steadv-stateis:
(Q+Q.)x = Q.X. (4.1)
Equation 4.1 states that the rate at which SS are
applied to the secondary clarifier (Q + Q,)X is equal to
the rate at which they are removed.(QJ,).The equation
can be used to develop the following useful relationships
for evaluating secondaryclarifier operation:
a. Degree of Thickening Achieved
by Secondary Clarifier
x,lx=(O+O,)/0, (4.2)
This relationship shows that the degreeof thickening is a
function of the influent flow and RAS flow rates. Within
the limits described below, the degree of thickening is
determined by the plant operator by selecting a particular
value of Q.
b. Required RAS Flow Rate
Q. = QX(X, -X) (4.3)
This relationship can be used to calculate the RAS flow
rate required for a specified influent flow rate, specified
aerationbasin MLSS concentration,and RAS SS concen-
tration believed (perhapsbasedon history) to be achiev-
able in the secondaryclarifier.
c. Secondary Clarifier Capacity
e =e,(x,-x)/x (4.4)
This relationshipcan be used to calculatethe capacityof
a secondaryclarifier for specifiedvaluesofRAS flow rate,
aerationbasin MLSS concentration,and achievableRAS
SS concentration.Equation 4.1 to Equation 4.4 assume
that sufficient secondaryclarifier surface area is available
to thicken the RAS to a concentration of X. Techniques
for determining the secondary clarifier surface area
requiredto achievethis are discussedin the next section.
4. SludgeThickeningTheory
Thickening of activated sludge SS in a secondaryclarifier
occurs by gravity settling. As the SS settle, their concen-
tration increasesfrom the MLSS concentration (X) to the
RAS concentration(X.). Becauseof their flocculent nature
and concentration, activated sludge particles do not settle
individually; rather,they settle as an entire mass.The mass
of SS settlesat a constantrate until it begins to accumulate
at the bottom of the settling vessel. The initial settling
velocity of the activated sludge SS (Vt) decreasesas the
initial SS concentration increases.This type of behavior
is called zone or Type III settling.
Zone settling is familiar to anyone who has run an
SVI test. The entire mass of activated sludse SS settles
BO
M anua lo n C a u s e sa n d C o n tro lo f A c ti v atedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
1
.:o
Hisher initial concentration
E unit of secondaryclarifier surface area. Using the defini-
0 Time --------->
sludgesuspended
FfCURE4.2 Zonesettlingof activated solids.
together,producing a well defined interfacebetweenthe
top of the settling sludge and the clear liquid above it.
When the height of the interfaceis plotted againsttime,
the line initially is straight but later begins to level off
(Figure 4.2). The slope of the initial straight line is the
initial settling velocity V,. Activated sludge with higher
initial SS concentrationssettlesslower than sludgewith a
lower initial SS concentration(Figure 4.2).
Secondaryclarifier thickening capacity is related to
the ability of the clarifier to accept the applied SS load
and to convey all of the SS to the underflow (RAS line).
This is accomplished by gravity settling of the activated
sludgeSS and withdrawal of RAS by pumping it back to
the aerationbasin.RAS withdrawal causesa generalflow
of fluid to the bottom of the secondaryclarifier, and this
generalflow also carriesthe settling activatedsludge SS
to the bottom where they can be removed.
Severaltechniqueshave been developedto determine
the thickening capacities of secondary clarifiers. Gener-
ally, the rate at which activated sludge SS are applied to
the clarifier is compared to the rate at which they are
conveyed to the bottom of the clarifier by settling and
withdrawal. As long as the SS application rate is not
greater than the SS settling rate plus the SS withdrawal
rate, the applied SS will be conveyedto the bottom of the
secondary clarifier and removed (i.e., thickening failure
will not occur). If the SS application rate is greater than
the SS settling rate plus the SS withdrawal rate, excess
SS will accumulatein the secondaryclarifier until it is
full and then SS will overflow into the secondaryeffluent.
P r oblem s
Techniquesfor secondaryclarifier thickening analysis
are relatively complex. They accountfor SS concentration
increasesin the secondaryclarifier during settling and for
the effects of the increases on the rate at which SS are
conveyedto the bottom of the secondaryclarifier through
settling and withdrawal. These techniques are discussed
thoroughly in the literature (Keinath et a1.,7977 Ekama
et al., 199'7;Wahlberg,2001). They will not be discussed
in detail here; rather we will present a simple approach.
The thickening capacity of a secondary clarifier can
be statedin terms of an allowable applied SS loading rate
G, which is the mass of SS applied per unit of time per
tions presentedearlier,the actual SS loading rate is:
c" = X(e+a,)/A (4.s)
Comparison of the actual SS loading rate with the
allowable SS loading rate will indicate whether the sec-
ondary clarifier is overloadedand is likely to experience
thickening failure. The allowable SS loading rate is a
function of several factors including the RAS flow rate
per unit of clarifier surface area (Q,/A) and the settling
characteristicsof the activatedsludge.By varying these
factors,it may be possibleto increasethe SS thickening
capacity of the secondary clarifier and the resulting SS
holding capacityof the activatedsludge system.
Secondary clarifier thickening capacity can be deter-
mined by direct measurementor by calculation. Direct
measurementof thickening capacity involves manipula-
tion of the secondaryclarifier SS loading until thickening
failure occurs.For example,the secondaryclarifier influ-
ent flow rate can be increasedwhile the RAS flow rate is
maintainedconstant.The secondaryclarifier sludgeblan-
ket depth is monitoredand the point at which it beginsto
increasecorrespondsto the start ofthickening failure. The
applied SS loading rate under these conditions is calcu-
lated. This is the allowable SS loading rate for the selected
RAS rate and the observedSS settlingcharacteristics. By
repeating this procedure at different RAS rates and with
activated sludge of different settling characteristics, the
secondaryclarifier thickeningcapacitycan be established
for a range of conditions.
This technique has been used in practice, but it is
cumbersome and time-consuming. Also secondary efflu-
ent quality may be compromised during testing if the test
is not terminated before the sludge blanket rises too far.
Moreover, bulking sludge must be available to allow the
secondaryclarifier SS loading capacity to be measuredfor
bulking conditions. The major advantageof this method
is that the allowable SS thickening capacity measurements
are direct and no assumptionsare required to translatethe
results to full-scale plant performance. The method can
 Control of ActivatedSludgeBulkingand Other SettlingProblems
400
50mL/yz\
300
a
a . 12mld,(-300 gpd/ftr)
F 1sn
ra
E 200
I
I rso
d
B L \t
€ roo
50
l0 15
UnderflowTSSconcentration,
FfGURE 4.3 Clarifier operatingdiagram for unstirredSVI. (Adapted from Daigger,G.T. (1995), WaterEnviron. Res.,67,95.)
be applied retroactively to the analysis of a sludge bulking
incident that leads to secondary clarifier SS thickening
failure. Determination of the secondaryclarifier SS load-
ings that resultedin thickening failure yields valuesthat
can be used to develop operating strategiesthat avoid it
in subsequentbulking incidents.
Calculationtechniquesfor determiningallowablesec-
L- ondary clarifier SS thickening capacity require the mea-
surementof SS settling characteristics.The relationship
betweenSS concentrationand the initial settling velocity
must be established,either by direct measurementor by
using previouslydevelopedcorrelationsbetweenan index
of sludge settleability and the SS concentration/initial set-
tling velocity relationship.Correlationshave been devel-
oped using sludge settling indices such as the standard
(unstined) SVI, stined SVI conductedat MLSS = 3.5 glL
(SSVI.5),and diluted SVI (DSVD.
Direct measurement of the SS concentration/initial
settling velocity relationship can be cumbersome and
time-consuming.Initial settling velocities must be mea-
suredfor activatedsludgewith a range of initial SS con-
centrations obtained by mixing various proportions of
mixed liquog RAS, and secondaryeffluent. Measurements
must be made in settling columns at least 0.15 m (6 in.)
in diameter, 1.8 m (6 ft) high, and equipped with stirring
devices.Bulking sludge must be availablebefore its thick-
ening characteristicscan be measured.The resulting SS
concentration/initial settling velocity relationship is then
used to determine the allowable clarifier SS loading rate
under a variety of operating conditions using either ana-
lytical expressions(Baskin and Suidan,1985)orgraphical
techniques such as state point analysis (Keinath et al.,
81
. 20 mld (-500 gpd/ft2)
. l6rnld(-400gpdlft2)
50€
U)
U)
F
4( l
E
JU E
o
='
l0
(-50 gpd/ft2)
20 30 35
g/L
1917; Ekama et al., 19971, Wahlberg,2001). State point
analysishasbeenusedprincipallyby engineersfor design-
ing activatedsludgesystemsbut its use by plant operators
is incqeasing.Using the analysisto evaluatethe impacts
of sludge thickening characteristicson activatedsludge
plant capacityrequiresthe developmentof SS concentra-
tion/initial setting velocity relationships for activated
sludge with both good and poor settling characteristics.
Correlations such as those discussedabove can be
coupledwith standardthickeningcapacitycalculationsto
producea secondaryclarifier operatingdiagram.Figure 4.3
presentsthe results of one such correlation developedby
Daigger (1995).The allowableSS loading rate G, is plot-
ted as a function of the RAS SS concentrationfor activated
sludge with standardSVIs (unstirred in a l-L graduate
cylinder) in the range of 50 to 350 ml/g. Dashed lines
that correspond to various underflow (RAS) rates (Q"/A)
also are plotted.Similar secondaryclarifier operatingdia-
grams for SSVI35and DSVI are presentedin Figures 4.4
and 4.5, respectively.
To use the secondaryclarifier operating diagram, the
point that correspondsto the clarifier operating conditions
(operating point) is located using two of the following
pieces of information:
. Actual SS loading rate (G)
. Underflow (RAS) rate (Q"/A)
. RAS SS concentration(X")
If all these data are available, then the third piece of
information can be used as a check. When the operating
point plots below the line corresponding to the current
82 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

50 ml-/g 20 m/d (-500 gpd/ft'?)

E
60 ml-/g . 16 m/d (-400 gpd/ft2) E
100

U) (n
a U)
F rsn F

40 -l
{
tr 200 8 m/d (-200gpd/ft2)
! !

o
3 rso
B rn= o
: 1oo
2 rnld(-50 epdfit2)
10

15

Underflow TSS concentration, g/L

F |G URE 4. 4Cla ri fl e ro p e ra ti n g d i a g ra m fo rSS V l rr.(A daptedfromD ai gger,G.T.(1995),W aterE nvi ron . Res. , 67, 95. )

50 ml-/g 20 tr/d (-500 gpd/ft,)

16mld(-400gpd/ft,)
60 ml-/g ,. - !
o" 300 -
E 508
. 12nld (-300gpd/ft,) U)
(h
U) U)
r LJU F

40 1
100ml-/g
€ 2oo , 8 m/d (-200 gpd/ft2)
-
= 30€
3 rso o

I
B
r^^ - -
4 n/d (-100 gpd/ft2) 20e
2 ttld (-50 gpdlft2)
t0

t0 15
Underflow TSS concentration, g/L

FICURE 4.5 Clarifier operating diagram for DSVI. (Adapted from Daigger, G.T. (1995), Water Environ. Res., 67,95.)

SVI, the clarifier SS loading rate is less than the limiting
clarifier SS loading rate and the secondary clarifier is
operating below its allowable SS thickening capacity.The
clarifier should not be subject to SS thickening failure.
When the operating point falls directly on the line corre-
sponding to the current SVI, the clarifier SS loading rate
equals the limiting clarifier SS loading rate and the sec-
ondary clarifier is operating at its failure point. When the
operating point plots above the line corresponding to the
current SVI, the actual clarifier SS loading rate exceeds
the limiting clarifier SS loading rate and the secondary
clarifier is operating above its allowable SS loading. In
this caseclarifier thickening failure will occur.An example
of the use of this diagram appearslater in this section.
Correlations such as those presented in Figure 4.3
through Figure 4.5 were developed by several workers
(White, 1976; Johnstone et al., 1979; Koopman and
Cadee, 1983; Pitman, et al., 1983; Daigger and Ropeq
 Control of ActivatedSludgeBulkingand Other SettlingProblems 83

TABLE4.1
Comparisonof SludgeSettleabilityIndices
lndex Settling Device Low-Speed Stirring SS Concentration

Conventional SVI (SVI)
Mallory SVI (SVI.)
Diluted SVI (DSVI)
Stirred SVI at 3.5 g SS/L (SSVI3.5)
l-L graduate cylinder
Mallory Settleometer
1-L graduatecylinder
1-L graduate cylinder
No Aeration basin mixed liquor
No Aeration basin mixed liquor
No Mixed liquor diluted so that settled volume in
graduate cylinder is <200 mLlL
Yes Tests at several SS concentrations; value at 3.5 g/L
obtained by interpolation

1985; Wahlberg and Keinath, 1988; Daigger, 1995).All

Q,= QX (X ._X )
thesecorrelationswere applied successfullyto the analysis
of secondaryclarifiers at full-scale wastewatertreatment _ 3000)
= (1.0)(3000)/(12,000
plants.Two things must be rememberedwhen using any
of these correlations. First, because all correlations are = 0.33MGD (rZS O.,/O)
empirical,they have limited rangesof application.When-
ever possible the correlation used in a particular situation

t_ should be "calibrated" by comparing actual secondary

clarifier SS thickening pedormance with predictions of
the empirical correlation.Second,the various indices of
activated sludge settleability used in the correlations are
not equivalent.Table 4. I summarizesthe characteristics
of the four most commonly used settleability indices.
While DSVI and SSVI* seemto provide the most accu-
rate estimatesof activatedsludge settling characteristics,
it is generallycurrent practicein the U.S. to use the stan-
dard unstined SVI conductedin a l-L graduatecylinder
becauseof its simplicity. The standardunstirred SVI is
used in the examplespresentedin this chapter.
Correlations between activated sludge settleability
indices and settling characteristicshave been used suc-
cessfully to analyze the behavior of full-scale activated
sludge plants.Although care should always be exercised
and the relationshipsshould be calibratedwheneverpos-
sible, they can be used with confidenceto estimatesec-
ondary clarifier SS thickening capacity.
5. Secondary Clarifier Analysis and Operation
The first stepsin secondaryclarifier analysis are using the
relationshipsin Equation 4.1 through Equation 4.4. For
example,sludgebulking generally will causea decrease
in RAS SS concentration. Equation 4. I predicts that a
decrease in RAS SS concentration (X,) requires an
increasein RAS flow rate. This increaseis necessaryto
ensure that SS applied to the secondary clarifier are
removed in the RAS.
For example, consider an activatedsludge system that
operatesat an influent flow rate of 1.0 MGD (3785 m3/d)
and an RAS flow rate of 0.33 MGD (1250 m3/d). The
MLSS concentration is 3000 mg/L and the RAS SS con-
centration is 12,000 mg/L. As a check, calculate the
required RAS flow rate using Equation4.1.
This agrees with the actual RAS flow rate. Now
assumethat sludge settleability deterioratesand secondary
clarifier sludge blanket depth begins to increase.The RAS
SS concentrationis found to be 8000 mg/L. Equation4. I
indicatesthe RAS flow rate must be:
. Q.= QX (X ,_X )
= (l .0)(3000)/(8000- 3000)
= 0.60 MGD (ZZtOm3 la)
Thus, for these conditions increasingthe RAS flow
rateto 0.60 N.IGD(2270m3ld)will be necessaryto prevent
secondaryclarifier SS thickeningfailure.The useof Equa-
tion 4.1 to Equation4.4 to developalternativesecondary
clarifier operating strategiesmust be accompaniedby an
analysis of secondaryclarifier thickening capacity.The
correlationmethod (with the standardunstirred l-L SVI)
of Daigger (1995) will be used to illustrate this analysis.
Assume that the secondary clarifier in the example
discussedabove has a diameter of 45 ft (13.7 m) and a
surfaceareaof 1590 fP (148 m2).From Equation4.5, the
applied SS loading rate for the initial operatingcondition
i s:
c" = X(e*e,)/A
= (3000)(1.0
+ 0.33)(8.34)/(15e0 )
= 20.9rblf(, d (102kg/mr, d)
This condition plots as point I in Figure 4.6 and indi-
cates that the secondary clarifier could operate success-
fully with a sludgehaving an SVI of up to 150 ml/g.
84 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

80 In I I
390
50 mUs \ i+-,orooo)d/ft')
v \{ro-o 00 gpd/ft'z)
340
70

' \<-
x,,
I 6nld (400 gpd/ft'?.
N
,i 60 /, ros {
100mUg c'
o
a
U) :-* l2mld (300 pd/ft2r
9so ) 45 =

bn
,!y4 X
o 40 . 1o< -
,/\ oa
d (200 gpd/ft'z)
200 ,Ur4 , \ ,/ \,, (h
mglrl \
8,zoo -l
(,
30.'7 _-AY' \-" r45 >
o3 0
-, ) 5 ' 250 mLlt
ru m/d (lm gpd/ft'?)
o 3rJOmLlg/ 4 )d $_-"-
"'^yt
"n \_-- \ 100E
-)n l\
YN
\ B
o
r
l0
\ \-F
,..\ |
r nn ,"}e\ -.\ 6-- -

/
/' /r /

, aJl
../ -6.31
- )9
\Hi \-
I
\-.-
-'2,.4-, tl tl
gpAft'?)
Zmla-(SO

05 1 0 l5 25 30
RAS SS concentrationX. gSS/L

clarifierthickeninganalysis.
FIGURE4.6 Exampleof secondary

For the secondoperatingcondition (RAS SS = 8000 rate of 591 gpd/ft2 (24.4 mld), but that the SVI will be
mg/L, RAS flow rate = 0.6 MGD) the applied SS loading controlled to a value of no more than 150 ml-/g by RAS
rate is: chlorination (see later sections in this chapter). Moving
along the operating line (for 597 gpd/ft2) to point 4 in
(3ooo)(1.0 =
+ 0.6)(8.34)/(15e0) Figure 4.6 indicates that the secondaryclarifier could be
operated at an SS loading rate of up to 42.8 lb/ftzld
o (tzr tg/m'z,0)
25.2tblft2, (209kglm2/d) and with an RAS SS concentration of up
to 8700 mg/L.
This condition plots as point 2 in Figure 4.6 and indi- Equation 4.5 can be rearrangedto calculate allowable
catesthat the secondaryclarifier now can operatesuccess- influent flow rate and the allowable MLSS concentration
fully with a sludge having an SVI of just over 200 mL/g. for theseconditions. To calculate the maximum allowable
Now assumethat the maximum RAS pumping capac- influent flow rate at MLSS = 3000 mg/L:
ity is 0.95 MGD (3600 m3/d).At the maximum RAS flow
rate, the bulk withdrawal rate is 0.95/1590 or 591 gpd/fe
Q = (c" A /X )_ Q.
(24.4 mld), and the applied SS loading rate is:
= (43.s)(r - 0.es
se0)/((3000)(8.34))
=
+ 0.e5)(8.34)/(1se0)
(3000)(1.0
= 1.9 MGD (ltSO mt ld\
30.7lb/ft',d(tsot g/rn',a)
To calculate the maximum allowable MLSS at an
This condition plots as point 3 on Figure 4.6 and influent flow of 1.0 MGD (3785 m3/d):
indicates that the secondary clarifier could be operated
successfully with a sludge having an SVI of about 250 x=(c"e)/(e+e,)
mllg and that the RAS SS concentration would be about
6300 mg/L. = (43.s)(1se0)/(1.0
+ 0.esX8.34)
Now assumethat the RAS flow rate is maintained at
0.95 MGD (3600 m3/d), equivalent to a bulk withdrawal
=4250 mglL
 Control of ActivatedSludgeBulkingand Other SettlingProblems
These examples illustrate how the general principles
of secondary clarifier analysis can be used to optimize
existing operation.
6. System Analysis and Operation
In addition to manipulation of RAS flow rates, the effects
of bulking sludge can be ameliorated by reducing MLSS
concentrationin the secondaryclarifier feed. This reduces
the SS load to the secondaryclarifier (Equation4.5).Also,
when the applied SS loading rate is reduced while main-
taining the same RAS bulk withdrawal rate, the allowable
SVI is increased(i.e., moves downward and to the left
along an RAS bulk withdrawal operatingline in Figure 4.6).
The SS concentration in the secondaryclarifier feed
can be reduced by reducing the MLSS inventory or by
changing the activated sludge operating mode. MLSS
inventory can be reduced by increasing the activated
sludge wasting rate. This is only feasible if sludge han-
dling capacity is available and if reducing the MLSS
inventory does not unfavorably affect the FlN4 (e.g., low
F/I\4 may be required to achieve nitrification).
The objective of changing operating mode to combat
sludgebulking is to reducethe MLSS concentrationin the
l-. secondaryclarifier feed without reducing the MLSS inven-
tory. Aeration basins incorporating step feed (step aera-
tion) and contact stabilization configurations are particu-
larly useful in this regard.Both configurationsallow the
operation of the first part of the aeration basin at a higher
MLSS concentration,and the last part of the aerationbasin
at a lower MLSS concentration,than when the inlluent
0.33MGD. X,.= 12.000mg/L
"Plug flow" (conventional)mode
0.5 MGD
z = 2370
1 . 33MGD .2310
0.33MGD.X-=9230
Step feed mode
FIGURE 4.7 Schematicdiagram of step feed operation.
B5
wastewater and the RAS are both fed to the head end of
the aeration basin. Total SS inventory does not changefor
a given F/M. The SS concentrationentering the secondary
clarifier, and therefore the SS loading rate, both decrease
compared to a system in which the influent wastewater
and RAS are both fed to the headend of the aerationbasin.
The use of step feed to lower the SS loading applied
to the secondary clarifier will be illustrated by an exten-
sion of the previous example. In addition to an influent
flow rate (Q) of 1.0 MGD (3785 m3/d),an RAS flow rate
(Q,) of 0.33 MGD (1250 m3/d),an MLSS concenrrarion
(X) of 3000 mg/L, an RAS, SS concentration(X,) of
12,000 mg/L, and a secondaryclarifier SS loading rate
(G") of 20.9 lbfir2d (102 kg/m2d), it is assumedthat the
aerationbasinhasa total volume (V) of 0.25 MG (950 m3)
and can be operatedin a completelymixed mode or in a
two-compartment(pass)step feed mode (Figure 4.7).
For the completely mixed mode, point one in Figure
4.8 indicatesthat the plant can be operatedsatisfactorily
undertheseconditionswhen the SVI is lessthan 150ml/g.
When the operation is changedto a two-compartmentstep
feed mode at the same influent and RAS flow rates,all the
RAS and only one-half of the influent flow are added to
the first compartment.The remainderof the influent flow
is added to the secondcompartment.A redistributionof
MLSS inventory occurs so that the first compartmentcon-
tainsmore MLSS than the secondcompartment.This hap-
pens becauseless dilution of the RAS by influent occurs
in the first compartment.If the total MLSS inventory
remainsthe same as in the completely mixed mode, the
Gu = 20'9 lb/ft2, d
Gu= 16.1lbfie, d
B6 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

80 ttl t^tl
-390
50 ml-/g \oiroruoot dlft')

\40"u0, )0 gpd/ft2)
-340
70
/\/

c 60 2\/
vo 1mld(400 gpd/ft2
-295 €
100mlig
c!
F X/ @

(h
;'z-J2nld (300 pdtftz)
?s o -245
./\ / @

,i mnrA4/ /\,
|<
l, qo ,/\ -195 ll
oq
je4 t , /V (200 spdlfr2)
a
200n
(RAS= 0.9sN ;ot/3V' i=q,6/r4cD)
,,/\
208gpd/ft2
-i(\* -r45
i]

q)
v)
#:o
B
=
4zo
250 mLlp
300mllg,
r&
++J)
.\K:,
33I\,TGD)
\ rd(100gpd/ft2)
\-[- ...- -100

GN
-350mL/s. / ,/

1*17 aX\r X

;z =,>s
-\ (t-
t0 - -50
n/d (i0 gpd/ft2)

rltl
tltl
-0
0510 t5 20 25 30
RAS SS concentration,X' gSS/L

FIGURE4.8 Exampleof stepfeedoperation.

MLSS concentration in the second compartment will be =(0.83

(0.83Xx,) +o.s)(x,)
lower than when the system was operated in the com- (4.7)
pletely mixed mode. = (0.625)xr
x, = (0.83/1.33)xr
The MLSS concentrationsin eachof the two compart-
ments can be calculated for the step feed mode by writing Combining Equations(4.6) and (4.7) gives:
an SS massbalanceequation for eachcompartment.These
equationsare then substitutedinto an equation for the total Xr = (0'625)X,
SS inventory, which is solved for the SS concentrations (+.g)
in each compartment and in the RAS. For our example, = (0.2s)x,
= (0.62s)(0.4)x.
the aeration basin MLSS inventory in the completely
mixed mode is: The total aeration basin MLSS inventory can be
expressedas a function of X. as shown below. Remember
= 62sstb (2840kg)
(3000X0.2s)(8.34) that the volume of each aeration basin compartment is
0.25 ll4clz or 0,125 MG (473 m3).
This SS inventory will be maintained in the two-com- Aeration basin suspendedsolids inventory:
partment step feed mode. The SS mass balance for the
point at which the wastewater influent enters the first (x.)10.r2s11a.34) =
+(x,X0.12s)(8.34)
compartment (0.5 MGD) and is mixed with the RAS flow
(0.33 MGD) is: (1.0425Xx,
+xr)

= (0.s+ 0.33Xx1)
(0.33Xx,) Substituting for X, and X, from Equation 4.8 gives
A.6\ total aeration basin SS inventory:
= (0.4)x.
x, = (0.33/0.83)x,
(1.0425X0.4x. = (0.677
+ 0.25X,) 6)x,
The SS mass balance for the point at which the flow
from the first pass(0.83 MGD) mixes with the wastewater Setting this equal to the total MLSS inventory of
influent flow to the second compartment (0.5 MGD) is: 6255 lb gives:
Cont r olof A c t i v a te dS l u d g eB u l k i n ga n d O th e r S ettl i ngP robl ems
6225 tb = \0.6776)X,
X,= 9 2 3 0 mg l L
X, can be calculatedusing Equation4.6
X, = (0.4)X. = (0.4X9230)
=3690 mglL
X, can be calculatedusing Equation4.8
X, = (0.25)X,= (0.25X9230)
=23t0 mglL
The secondaryclarifier SS loading rate for this oper-
ating condition is:
6" = (2 3 1 0 )(1 .3 3 )(8 .3 4 )/(1 s e o )
= t6 .r tb l ftz ,d (7 8 .6 tg /m" o )
This is plotted as point four in Figure 4.8.
The benefits of two-compartment step feed operation
in allowing an activatedsludgesystemto operatesuccess-
fully with a poor settling sludgeare illustratedin Figure 4.8
using the data from the previously calculatedexamples.
Point one through point three are for completely mixed
operationswith RAS flow rates of 0.33, 0.6, and 0.95
MGD (1250,2270, and 3600 m3ld),respectively. Point
four is for two-compartment step feed operation at an RAS
flow rate of 0.33 MGD (1250 m3/d),as calculatedabove.
These operating points show that an activatedsludge with
an SVI of approximately 220 to 230 ml-/g requires an
RAS flow rate of 0.95 MGD (3600 m3/d) when the aera-
tion basin is operatedin the completely mixed mode but
only 0.33 MGD (1250 m3/d) when the aerationbasin is
operated in the two-compartment step feed mode. This
resultsin lower RAS pumping costs and higher RAS SS
concentration (9230 mg/I- vs. 6300 mg/L) - which may
reducewaste sludgeprocessingand disposalcosts.
Two-compartment step feed is the simplest alternate
operating mode. Operation in other step modes (such as
three- or four-compartment), contact stabilization, or step
feed with RAS reaeration(a combination of step feed and
contact stabilization) offers greateradvantageswith regard
to reducing secondaryclarifier SS loading rates to accom-
modate a poor settling sludge. Indeed, any change that
resultsin lessdilution of RAS by influent wastewaterwill
allow storageof activated sludge SS in the aeration basin
and decreasethe SS concentrationofthe secondaryclarifier
feed. For example,if the systemdepictedin Figure 4.8 had
been placed in the contact stabilization mode, using the
first compartment for sludge stabilization and the second
87
compartment for contact (while maintaining the same
MLSS inventory),the SS concentrationin the secondary
clarifier feed would havebeen reducedto 1200 mg/L and
the secondary clarifier SS loading rate would have been
8.4 lb/ft2d (41 kg/m2,d).
These examplesillustrate calculation proceduresfor
determining operating conditions when a mode of opera-
tion is changed.They can be simplified for a particular
system and set of operating conditions or can form the
basesfor operatingdiagramsor spreadsheets. They also
can be usedas a guide to evaluatethe effectsof operating
changes.A generalprinciple is that changesresulting in
less dilution of RAS with influent wastewater will
decreasesecondaryclarifier feed SS concentration.Redis-
tribution of SS usually takes less than I d, so the results
can be seenvery quickly following changesin operating
mode.
In addition to dealingwith bulking sludge,manipula-
tion of aerationbasin wastewaterand RAS feed points is
often used to handle high wet weather flows. The solids
loading rate of a secondaryclarifier increasesas influent
flow increases.Many plants "step the feed down the aer-
ation basin" (convertfrom conventionaloperationtoward
contactstabilization)as the wastewaterflow rateincreases.
This type of action can prevent solids washout during
storm .flows,The shorteraerationtimes afforded by con-
tact stabilizationare offset somewhatby the dilution of
wastewaterduring storm flows.
B. AoorrroN oF CHEMrcArs Solros ro
AND INERT
ErunnNcr
AcnvnrroSluocrSrrnrxc Rlrrs
Severalapproachesfor improving activatedsludgesettle-
ability fall into this category in which settleability is
improvedwithout eliminatingthe root causeof a problem.
Syntheticpolymerscan be addedto municipal and indus-
trial activatedsludge plants to overcomethe bridging or
diffuse floc structuresassociatedwith excessivefilamen-
tous organismgrowth and flocculatedispersedflocs and
microorganisms.Polymers also have been used to aid
settling of activatedsludge containing large amounts of
water-retentiveextracellularmaterial (viscousor nonfila-
mentousbulking). Polymers have been added to control
activatedsludgefoaming due to nocardioformorganisms
(seeChapter5).
The use of synthetic organic polymers does not
increasesludge mass. This is their principal advantage
over inorganic coagulantsand flocculantssuch as alumi-
num and iron salts.Wide variationsin polymer type and
dose have been reported.In general,for bulking control,
a high molecular weight, high cationic charge polymer
alone or in combinationwith an anionic polymer may be
used.It is wise to perform frequentjar testing to determine
polymer dose,becausethe dosecan changefrom batch to
batch and with changesin activatedsludge properties and
B8 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
400
\
4 J
400
{ increasedthe total sludgeinventory l7%o,from 145,000lb
€o
P rnn
s -)(A
'
a were reduced. The beneficial effects of the 3 d of alum
! =
a dosing were maintainedfor about 3 wk.
r 00
02468 1 0
Polymer
Concentration,
mg/L
FIGURE4.9 Effectof a cationicpolymeron theSVI of a bulk-
ing sludge.(From Singer,P.C.et al. (1968),"/. WaterPollut.
ControlFed.,40,Part2, Rl. With permission.)
filamentousorganismtypes.Overdosingcan lead to dete-
rioration in performance;prolonged excessivedosescan
cause gelling of the activatedsludge so that its settling
properties deteriorate instead of improve. Figure 4.9
showsthe resultsof a typical jar test.Thesedata suggest
that a polymer dose of about2 mg/L would be appropriate
for the sludgebeing tested.
Polymersusually are addedto the mixed liquor as it
leavesthe aerationbasin or to the secondaryclarifier cen-
ter well. In general,the long-term addition of polymers
for bulking control is expensivecomparedto RAS chlo-
rination. Chemical costs for polymer addition are in the
range$10 to 50/MG comparedto $l to l0 for RAS chlo-
rination. An appropriateuse of polymer addition is dealing
with a settling problem on an emergency basis. Polymer
addition facilities can be installed quickly (for a solution
polymer, a storage tank, dilution water, piping, and a
metering pump are required). Settling improvement is
immediate. Typically, RAS chlorination takes a few days
to improve settling and the installation of facilities to use
it regularly is somewhat more complex.
Inorganic coagulants and flocculants such as ferric
chloride and alum (aluminum sulfate) have been used to
aid in activated sludge settling. Reductions in SVI were
notedby Eberhartand Nesbitt (1968) andZenz and Pivnicka
(1969) when alum was added to activated sludge for the
simultaneousprecipitationof phosphate.Thesechemicals
produce precipitates that sweep down a bulking activated
sludge or the small particles causing dispersedgrowth.
Finger (1973) used short-term alum addition at the effluent
end of the Renton (Seattle,WA) activated sludge plant to
reduce SVI from about 130 mllg to about 90 ml/g,
increase sludge bulk density, and increase settling rates
during periods of high (wet weather)influent flow. On one
occasion,alum was added for 3 d at a dose of approxi-
E
mately 60 mg/L based on wastewater flow. This dose
E
(65,800kg) to 170,000Ib (17,200 kg). Becausethe sludge
AJ
:; density increasedfrom 0.0075 giml to 0.0122 glmL (a
637oincrease),the secondaryclarifier sludgeblanket levels
An ATV Working Group report (1989) on European
practicescites the long-term use of ferrous sulfate(7 mg
as Fe/L) at the Wien-Semmering plant in Austria. Low
SVIs were obtainedin the presenceof abundantfilamen-
tous organisms.
Matsche (1982) reportedseveralexampleswhere the
addition of ferrous sulfate to the influent was successful
in controlling bulking. For example,at Neusiedl,Austria,
the addition of ferrous sulfate (10 to 14 mg as Fe/L) to
the influent of an activatedsludge plant treating canning
wastewaters reduced the SVI from 450 ml-/g to 60 to
70 mLlg and causedthe disappearance of Type 021N, the
filamentousorganismresponsiblefor the bulking.
Matsche (1982) points out that activatedsludgefrom
plants without primary clarifiers settlesbetter than that
from plantswith primary clarifiersbecauseof the presence
of the denserprimary SS in the activatedsludge.Matsche
also notes that dirt particles in sugar beet wastewater
improved activated sludge settleability. The PACT pro-
cess,(Hutton and Robertaccio,1975 and 1978) in which
powderedactivatedcarbon is addedto the aerationbasin
usually producesactivatedsludge that settleswell, even
in the presenceof significantfilamentousorganismlevels,
because of the weighting effect of the activated carbon
particles.Talc, a commercially available,finely divided
mineral, has been added to activated sludge plants in
France and the Netherlands to improve sludge settling
properties, but large increasesin the SS inventory occur
becausetalc to MLVSS ratios of approximately0.6 to 1.0
are required (Eikelboom and Grovenstein, 1998; Chudoba
and Pannier,1994).
It is common practice in some pulp and paper waste-
water activated sludge systems to intentionally waste
paper fibers or paper coating materials such as clay from
the mill to weight down poorly settling activated sludge.
The weighting action of inert biological SS has been
employed in activatedsludgeprocessmodificationssuch
as the Kraus process (Kraus, 1945) in which anaerobic
digester contents are recirculated through the aeration
basin system. While the addition of inorganic chemicals
Cont r olof A c t i v a te dSl u d g eB u l k i n ga n d O th e r S ettl i ngP robl ems
or inert biological solids can improve activated sludge
settling rates, it can impose significant additional solids
loads on the process.Moreover, the presenceof inert sol-
ids cannot always offset the adverseimpact of filamentous
organisms on activated sludge settling.
C. ro Srlrcnvtt-v Ktl.t"
Aoorrror.l oF DrsrNFEcrANTs
FrmmrrurousOncnuts,vs
1. G ener al
Chlorine and hydrogen peroxide have been used to kill
filamentous organisms selectively in activated sludge and
thus eliminatethe symptomsof bulking. Limited use has
been made of other disinfectants such as ozone (van Leu-
wen. 1988: van Leuwen and Pretorius.1998).Chlorine is
used in the form of a solution producedfrom a gas chlo-
rinator as sodiumhypochloritesolutionor as solid calcium
hypochlorite. Chlorination using a chlorine solution or
sodium hypochlorite solution is our choice for this type
of bulking control because:
. Chlorine is more economical than hydrogen
peroxide or ozone in the U.S.
. Chlorine and sodium hypochloritesolutionsare
much easierto dosein a controlledfashionthan
solid calcium hypochlorite.
. Secondary effluent disinfection of municipal
wastewater treatment plants is performed
mostly with chlorine or sodium hypochlorite
solutionsin the U.S.
Although ultraviolet disinfection is slowly replacing
chlorine for effluent disinfection, it will be many years
before it is a more widespread wastewater disinfection
process.Furthermore,the operatingstaffs at most waste-
water utilities are familiar with handling gaseouschlorine
and sodium hypochloritesolutions.For plants using chlo-
rine for effluent disinfection, the gaseousform or sodium
hypochlorite solution is available on-site and the addi-
tional amount required for bulking control usually is small
compared to the amount required for secondary effluent
disinfection. Even at plants not using chlorine for effluent
disinfection,some will be availableon-site for purposes
such as odor control, cleaning effluent filters, etc.
Certain situations may justify the use of materials
other than chlorine for bulking control, for example:
. Chlorine is not available on-site and cannot be
obtained immediately in sufficient quantities
when a severe bulking problem must be dealt
with.
. Chlorine is not used for effluent disinfection
and there is concern over the production of very
small amounts of chlorinated organics when
89
chlorine is added. This concern is often
expressedby petrochemical and pulp and paper
wastewater treatment plants. In a case history
presentedlater in this chapter, in-basin chlori-
nation of a plastics manufacturing waste acti-
vated sludge did not cause an increase in
effluent chlorinated organics. Because of the
fear of producing halogenated organic com-
pounds,chlorinationfor bulking control is rec-
ommended only as an emergency measure in
Germany and central Europe (ATV 1986).
. The proper applicationof chlorine for bulking
control does not interfere with BODr and SS
removal efficiencies.As one casestudy shows,
a small increasein effluent soluble COD. but
not effluent soluble BOD', occursduring chlo-
rination for bulkins control.
In one casewhere chlorination for bulking control was
practiced on a laboratory-scaleUniversity of Capetown
biological nutrientremoval system,nitrification,denitrifi-
cation, and phosphorus (P) release were not affected
(Lakay et al., 1988). Enhanced biological phosphorus
removal (EBPR) was adverselyaffectedby high chlorine
doses(8 kg Clrll03 kg MLSS/d) over a prolongedperiod
(19 d) but recoveredrapidly (within 5 d) when the chlo-
rination was stopped.
In a pilot plant study of the Virginia Initiative Process
(VIP) for EBPR, RAS chlorination for bulking control
with sodium hypochlorite at a dose of 2 kg Clrl103 kg
MLSS/d for 5 d reduced SVI from approximately 270
mL/g to 200 mL/g (Daiggeret al.,1988).Nitrification was
not affected but the effluent total P concentrations
increasedto values close to those in the influent. Poor
anaerobiczone P releaseand unusually high oxidation-
reductionpotential(ORP) were observedduring chlorina-
tion. Apparently, the chlorine interfered with the ability of
the P-accumulatingorganismsto releaseP in the anaerobic
zone and this further inhibited P uptakein the aerobiczone.
More typical anaerobic zone P release,ORP values, and
effluent P concentrationswere reestablishedalmost imme-
diately after RAS chlorination was discontinued.
In studieson a full-scale EBPR plant Sayman et al.
(1996) found that EBPR was not affected by a chlorine
doseof 0.7 kgClrll}3 kg MLSS/d. A moderateeffect was
observedat a dose of 1.4 kg Cl2ll}3 kg MLSS/d, and
EBPR was significantly affectedat a doseof 3 kg Clrl103 kg
MLSS/d. Theseresultssuggestthat RAS chlorinationcan
be used only at low chlorine doses for sludge bulking
control in EBPR plants.
Reports in the literature indicate that chlorination for
bulking control interfereswith nitrification (Wakefield and
Slim, 1988; Marstaller et al., 1992), but experiencesin
full-scale plants and the casehistories presentedhere show
that properly conducted chlorination for bulking control
90 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
does not affect the rate or extent of nitrification. The work
that indicated an effect on nitrification was conducted at
very low exposurefrequency (Wakefield and Slim, 1988)
or with batchwise chlorine addition (Marstaller et al.,
1992).Both of thesepracticesare inappropriatemethods
of using chlorine for bulking control.
Becausewe favor chlorination for bulking control, we
will deal first with this topic in detail and then presentsome
information on the use of hydrogen peroxide and ozone.
2. Use of Chlorination for Bulking Control
Until the 1970s,chlorination was used occasionallyfor
bulking control in the U.S. (Smith and Purdy, 1936;
Tapelshay,1945)but its use in the U.S. has becomewide-
spreadsince then. The ability to chlorinateRAS is often
designedinto new treatmentplants and frequently retro-
fitted into existing plants. Chlorination is used widely in
countries such as Germany (Frenzel and Safert, l97l;
Frenzel, 1977; Bode, 1983),Austria (Matsche, 1977),
Great Britain (Water ResearchCentre, 1983), and South
Africa (Wiechers,1983;Thirion, 1983).
Chlorine is popular in the U.S. becauseit is usually
availableon-site (since it is used for secondaryeffluent
disinfection) or can be acquired readily. The quantities
required for bulking control usually are small compared
with those required for secondaryeffluent disinfection;
sufficient chlorine is usually available to perform both
tasks.A common problem is controlling existing chlorine
delivery equipmentat the low rates required for bulking
control.
Two chlorinedosingpointsarecommon (Figure4.10):
directly into the aerationbasin and into the RAS stream.
The preferred dosing point is the RAS stream. Direct
dosing into the aerationbasin is used in plants with long
aeration basin hydraulic detention times, where dosing
chlorine in the RAS would not provide a sufficient fre-
quency of exposureof sludge inventory to chlorine or
when the RAS line is inaccessible.
Secondary
clarifier
(t)
vn
1J
Aeration basin
Waste activated
sludge
FIGURE 4.10 Chlorine dose points for bulking control.
Pr oblem s
a. ChlorinationCriteria
Several criteria must be followed for successfulbulking
control by chlorination. A target value of the SVI (or some
other activated sludge settling property) must be estab-
lished. The target SVI should be the value at or below
which the plant can be operated satisfactorily without
problems associatedwith poorly settling activated sludge.
Not only should the operation of the secondary clarifier
be considered in selecting a target SVI value, but the
impact of sludge settleabilityon solids processingunits
must be evaluated.Chlorinationshouldbe usedonly when
the SVI (or other settling property) target value is signif-
icantly and consistentlyexceeded.
A daily (or more frequent) trend plot of SVI values
should be made to determine when the target SVI is
approached.Sucha trendplot will aid in decidingwhether
a given SVI valuein excessof the targetvalueis consistent
with a trend or only an aberrant data point. Trend plots
can also assistan operatorin adjustingchlorine dosesto
anticipatechangesin SVI.
Chlorine solution must be addedin known and con-
trolled dosesto all of the activatedsludgeat a point where
excellentmixing is possible.Ideally,a separatechlorinator
(or feed pump for sodium hypochlorite solution) should
be dedicated to feeding chlorine for bulking control
becausethe dosesrequiredfor bulking control usually are
much lower than those employed for secondary effluent
disinfection. When retrofitting an existing plant to use
chlorine for bulking control, an independentlyvalved and
meteredchlorine solutionline can be taken off an effluent
chlorinatoror a hypochloritesolutionfeedpump.Accurate
knowledgeof the chlorine solution flow rate and concen-
tration in this line is imperative.An independentrotameter
in this chlorine solution line is mandatory.
As indicated,the preferredchlorine solution applica-
tion point is into the RAS line, althoughexceptionsexist.
If a plant has no common RAS line, the ability to chlori-
nateeachindividual RAS line must be provided.Sufficient
valving and metering must be supplied to allow control
and measurementof chlorine solution to each dosing
point.
Excellentinitial mixing of chlorine solution and RAS
is of the utmost importance.Reactionsof chlorine in RAS
are very rapid. Without excellent initial mixing, a large
part of the RAS might not be contacted by the chlorine
while a small part might be overdosed.One result of poor
initial mixing is the consumption of large amounts of
chlorine without control of bulking. Prolongeddosing at
a point of poor initial mixing can lead to the production
of turbid effluents (due to local over-chlorination) and a
reduction in treatment efficiency (due to excessivekilling
of floc-forming organisms).Examplesof poor RAS chlo-
rine injection points are RAS wet wells, mixed liquor
channels,and RAS or mixed liquor center wells of sec-
ondary clarifiers. The dose points of choice are locations
 Cont r olof A c t i v a te dS l u d g eB u l k i n ga n d Oth er S ettl i ngP robl ems
\.
)-
FICURE4.11 Examples of goodchlorinedosepointsfor
bulking control:(a) to the RAS line following an elbow,
(b) to the volute of an RAS pump, (c) to the riser of an
air-lift RAS system,and(d) to thedischargeof a traveling
bridgeclarifierRAS trough.
of high turbulence,for examplefollowing elbowsin pipes
( F igur e 4. lla) ; i n to th e v o l u te s o f R A S pumps
(Figure4.1lb); into their discharges;into and below the
rising liquid level in the riser tube of an airlift RAS pump
(Figure 4.tlc); and through a manifold into the waterfall
discharge to the aeration basin of an airlifUreturn on a
travelingbridge clarifier (Figure4. I 1d).If no other choice
is available, chlorine solution can be applied to RAS in
an open channel. It should be added at a point of turbu-
lence (e.g.,below a flow-measuringflume) and the injec-
tion should be multipoint through a pipe manifold or grid.
Because such devices can collect rags, they should be
designed to be removed periodically for cleaning.
When retrofitting an activated sludge plant for RAS
chlorination, it is imperative to carefully inspect the phys-
ical features of the system to identify suitable points for
91
chlorine injection. Chlorine should be added at a point
wherechlorine demandfrom wastewateris at a minimum.
Chlorine controlsbulking by disinfecting(killing) fil-
amentousorganisms.Any reactionswith wastewatercom-
ponentsthat modify or reducethe dosedchlorine concen-
tration will influenceits ability to kill the organisms.The
complexity and variability of wastewatermakes it impos-
sible to identify every reaction in which chlorine partici-
pates.However, the reactionsof chlorine with ammonia
and reduced inorganics such as nitrite and sulfide are
important.When chlorineis addedto a solutioncontaining
an excessof ammonia (ClrlN ratio < 5), it reactsrapidly
to form monochloramine (NHrCl).
Neethling et al. (1982) showedthat dosing activated
sludge solids with chlorine in the presenceof excess
ammonia is equivalent to dosing with monochloramine
92 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
J
o0
J^^
d zl)
$)
,=o
0 60 120
Time,s
FIGURE4.12 Chlorineresidualin RAS asa functionof initial
chlorinespecies,reactiontime, and the presenceof ammonia.
Initial chlorinedose= 2l mg Clr[L. SS = 2.77g/1-.Ammonia
concentration = 0.8mg NH,-N/L,.(a)and(b);ammoniaconcen-
tration= 11.3mg NH3-N/L,(c). (FromNeethling,J.B. et al.
( l 982),Report82-2,Sanitary
Engineering Environmental
Health
ResearchLaboratory,Universityof California,Berkeley.With
permission.)
(Figure 4.12). Monochloramineis not as potent a disin-
fectant as free chlorine, but it remains available longer
Thus, the presenceof ammonia does not materially affect
the bulking control ability of chlorine dosed to the RAS
unlessthe Cl, doseAllH.,-Nratio is high enoughto cause
the breakpointreaction(Cl2lNH3-N ratio of >10:l). If this
occurs.the chlorine will be reducedto chloride and less
chlorine will be availablefor attackingfilamentousorgan-
isms.The conditionsnecessaryfor the breakpointreaction
occur rarely and can be overcomeby increasingthe chlo-
rine dose.
The only common side reaction of chlorine that can
reduce its effectivenessin killing filamentousorganisms
for bulking control is with nitrite (NOr-). Chlorine reacts
rapidly and stoichiometrically with nitrite which can be
presentin activatedsludge (and RAS) during partial nitri-
fication. The HOCI + NO2- J NO3- + HCI reactioncon-
sumes5.1 mg Clrlmg NO2--N. The reactionis rapid and
consumes chlorine before it has a chance to react with
filamentousorganisms.
It is very unusual to encountersignificant levels of
sulfide in RAS. Thus, the consumptionof chlorine by the
oxidation of sulfide is not often important in RAS chlori-
nation for bulking control. We encounteredsulfide interfer-
encewith RAS chlorination only once.A broken secondary
clarifier sludge scraperallowed sludge to accumulateto a
point where sulfide levels consumed significant amounts
of chlorine as well as causing Thiothrix spp. bulking.
Chlorine reacts with the dissolved and particulate
organic matter present in wastewaters.The nature, rate,
and extent of these reactions have not been quantified
beyond saying that they are slower than reactionsbetween
chlorine and ammonia and between chlorine and nitrite.
Addition of chlorine to mixtures of RAS and influent
wastewater,such as those presentnear the head-endof an
aeration basin, should be avoided becausesome chlorine
may be consumedby reactionwith the high concentrations
of organicmatter (and possibly sulfide) and thus will not
be availablefor killing filamentousorganisms.High chlo-
rine doseswill be required for bulking control under these
conditions.Addition of chlorine to incoming wastewater
prior to the point where it mixes with RAS is completely
ineffective in killing filamentousorganismsand control-
ling bulking.
Chlorine dosesare measuredusually on the basis of
the activatedsludge SS inventory (overall mass dose in
kg Clrll03 kg SS inventory/d).The chlorine doseconcen-
tration at the dose point (mg Clrll-) and the frequency of
exposureof the SS inventory to chlorine (times/d) must
be consideredin choosinga dose point.
The most commonly used chlorine dose parameters
are concentration,overall mass dose rate, and frequency
of exposureof solids inventory to chlorine dose (Beebe
and Jenkins,198I ; Beebeet al., 1982',Jenkinset al., I 984).
All these parametersare expressedin terms of chlorine
doseratherthan chlorine residualbecausethe complexity
and variability of activatedsludgemake it difficult (if not
impossible)to predict the naturesand concentrationsof
chlorine residuals.This and the differencesin susceptibil-
ity of various filamentousorganismsto chlorine are the
reasonsfor significant variationsin the values of dosing
parametersfor successfulbulking control from one plant
to another and even from time to time at the same plant
(Neethlinget al., 1985a; 1985b).
Overall massdose(kg Clrll03kg SS/d)is a convenient
practical dosing parameter because it tells the operator
how much chlorine must be dosed over a 24-h period and
allows the dosing equipmentto be adjustedaccordingly.
This dosing parameter is related directly to the objective
of chlorination - to kill a fraction of the activated sludge
SS inventory each day. Daily organism kill is proportional
to the daily mass dose of chlorine (kg Clrld); systemSS
inventory is proportional to the total mass of organisms
in the system.
b. Ceneral Guidelines
The following rangesof overall mass dose rates are given
as generalguidelines:
Cont r olof A c t i v a te dS l u d g eB u l k i n ga n d Oth er S ettl i ngP robl ems
. 2 to 3 kg Clrl1O3kgSS/d- This overall mass
dose rate is typical of a maintenancedose that
may be effective when the SVI is generally
under control and continuous chlorination is
needed to kill newly growing filaments and
keep the SVI from increasing.
. 5 to 6 kg Clrl1O3kgSS/d - This is a typical
overall mass dose rate that will destroy excess
filaments and reduceSVI over severaldays with
little impact on effluent quality.
. 10 to l2kgClr/l03kg SS/d-This overallmass
dose will usually destroyexcessfilamentsand
reduce SVI very rapidly. Doses of this magni-
tude will also disrupt floc structure and result
in a deterioration of effluent quality.
Overall mass doses between the ranges cited above
will produceresultsintermediatebetweenthosedescribed.
c. Chlorination System Design
When designing a chlorination system, use the overall
massdoserate to determinethe size of the chlorine deliv-
ery equipment.In computing chlorine dose basedon the
sludgeinventory(kg Clrll03kg SS/d),the sludgeSS inven-
tory in the secondaryclarifiersshouldbe included.Where
two-basin processessuch as contact stabilizationare
employed,the sludgeSS inventoriesin both basinsand in
the secondaryclarifiers should be included in calculating
the overall chlorine massdose.
In most municipal wastewatertreatmentplants, the
RAS line is the best place to dose chlorine solution.The
minimum frequency of exposureshould be determined.
Frequenciesof exposure >3 times/d, generally are suffi-
cient. It may be possible to increasethe frequency of
exposureby increasingsludge recycle rates. Some RAS
chlorination systems(Bode, 1983) have operatedwell at
exposurefrequenciesas low as 1.4 times/d. Neethling et
al. (1985a)showedthat the requiredfrequencyofexposure
can be related to the relative srowth rates and efficiencies
"Domestic wastewater" case
a=
T = 6 mg Cl2lgSS,
C =7.5 mgClz/|,
f= 6 d-r
(a)
FIGURE 4.1 3 Chlorination parameters for: (a) typical municipal wastewater treatment plant and (b) long aeration time treatment
plant. T = overall mass dose rate; C = chlorine concentration at dose point; f = exposure frequency of sludge inventory to chlorine.
93
of kill of filamentous and floc-forming organisms.Simply
put, for chlorination to be successful, the filamentous
organisms must be killed faster than they grow in the
aerationbasin.
Activated sludge plants with long aeration basin
hydraulic detention times can have large SS inventories
so that daily SS flux in the RAS line is a small fraction
of the SS inventory.Becauseof this, a chlorine dosepoint
on the RAS line will provide a very low frequency of
exposure.Figure 4. 13 showsthe resultsof using a typical
overall chlorine mass dose rate for bulking control at
plantswith 4-d and 5-h hydraulicdetentiontimes (HRTs).
In the 5-h HRT activatedsludgeplant (Figure 4.13a),
the RAS flow rate is 50 m3/h (r = 0.5) and the RAS is
chlorinatedat an overall massdoserate of 6 kg Clrll03kg
MLSS/d. The chlorine dose concentrationis 7 .5 mg Cl.rll-
and the SS exposurefrequencyis 6 times/d.Thesecondi-
tions of chlorine concentration and exposure frequency
are satisfactory for bulking control. In the 4-d HRT acti-
vated sludge plant (Figure 4.13b), the RAS flow rate is
50 m3/h(r = 0.5) and the RAS is chlorinatedat an overall
mass dose rate of 6 kg Clrll03kg MLSS/d. The chlorine
dose is 121 mg C\rlL and the SS exposurefrequency is
0.37 times/d or once every 2.7 d. These conditions of
chlorine concentrationand exposure frequency are not
satisfactoryfor bulking control. A small part of the SS
inventory is exposed to very high chlorine doses. The
exposedsludge flocs will be destroyedand may increase
effluent turbidity. The SS inventory will not be exposed
frequentlyenoughto the chlorine doseto control filamen-
tous organism growth. Under these conditions, bulking
will continue and, in addition, a turbid effluent will be
produced. For such cases,a chlorination system using
severaldose points placed directly in the aerationbasin
(Figure4.10) shouldbe installed.
d. Monitoring Effects of Chlorine Addition
Reliablemeasurements of sludgesettling,effluentquality,
and sludgequality are requiredwhen chlorinationis used
"Industrial wastewater" (Long aelation time) case
0 =4 d
V. = 500m3
X" = 500mg/L
Chlorination
T = 6 mg Cl2lgSS,
C = 121mEClz/|,
f = 0.37d-r
(b)
94 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
for bulking control. The use of chlorine for bulking control
entails the purposeful addition of a disinfectant to a bio-
logical system.Becauseof this, frequent control teststo
assessthe effectsof chlorine addition must be performed.
Such control tests should include the following:
. A reliable and appropriate measurement of
sludgesettling such as SVI, DSVI, stirredSVI,
zone settling rate, or sludge blanket depth in
the secondary clarifiers at known wastewater
and RAS flow rates.
. A measure of secondaryeffluent turbidity. An
increasein secondaryeffluent turbidity accom-
panied by a rapid decreasein SVI indicates
chlorine overdose.Increasedsecondaryeffluent
turbidity indicatesthat significant amounts of
activatedsludgefloc havebeenbroken up by the
chlorine. Indeed, one observationcommonly
made when chlorine is applied improperly in
shock dosesfor bulking control is that the sec-
ondary effluent tums "milky" due to the presence
of small particles of broken-up activatedsludge
floc producedby excessivelyhigh chlorinecon-
centrations.Turbidity can be measuredwith a
nephelometeror can be appraisedvisually by
making transparencymeasurements directly in
the secondaryclarifier or chlorine contactcham-
ber with a Secchidisc (Cialdi, 1866).Transpar-
ency measurementsof secondaryclarifiers are
only possiblewhen sludgeblanketlevel is lower
than the extinctiondepth of the Secchidisc.
. Microscopic examination of the activated
sludge is very important when chlorination is
used for bulking control becausethe effectsof
chlorine on both the filamentousorganismsand
the flocs are readily observable.When chlori-
nation is beginningto exert effectson filamen-
tous organisms,it often is possible to see
progressivelyincreasingamountsof damageto
the cells and filaments.
The progressiveeffect of chlorinationfor bulking con-
trol on Thiothrix I, a sheathedfilament, is illustratedin
Figure 4.14.T\e healthy Thiothrix filament containing sul-
fur granulesis illustratedin Figure 4.14a.The first effect
of chlorinationis the disappearance of intracellularsulfur
granules that presumably are oxidized by the chlorine
(Figure 4.14b). The chlorine then progressivelycauses
cells to deform, detach from the sheath, and "ball-up"
(Figure 4.14c). Gaps then appearin the sheathsleft by the
disappearanceof cells (Figure 4.14d). Finally, empty and
broken sheathsremain (Figure 4.14e).
The initial effects of chlorine on filamentous organ-
isms described above can precede improvement in acti-
vated sludge settling properties and therefore are useful
Pr oblem s
as early signs that chlorinationis beginningto exert con-
trol over filamentousorganism growth. Theseobservations
allow the operator to determine when a satisfactory chlo-
rine dose has been reached.As a rough guide, satisfactory
filament control will be achieved when about 60 to l07o
of the filaments show observabledamage.For filamentous
organisms without sheaths, the SVI usually declines
within 1 to 2 d. For sheathedfilaments, the empty sheaths
must be washed out from the activated sludge system by
sludge wasting for the SVI to decrease.This can take a
longer period (up to I to 2 MCRTs).
Chlorine overdosingcan also be detectedmicroscop-
ically. The total elimination of filamentous organismsand
the presenceof small, broken-up flocs together with fine
particlesare signs of chlorine overdosing.
3. Case Histories of Bulking Control
Using Chlorination
a. Ceneral
Chlorinationhasbeenusedsuccessfullyto control bulking
causedby every type of filamentousorganism observed
in activated sludge. It is effective against filamentous
organismsthat causebulking by floc bridging and those
that causebulking by producingdiffuse flocs.Filamentous
organisms vary in their susceptibilities to chlorine. Thio-
thrix spp. and Type 02lN are quite susceptible;M, parvi-
cella and Nostocoida spp. are rather resistant, and the
remainderhaveintermediatesusceptibilities.Nonfilamen-
tous (viscous,zoogloeal)bulking is not controlledby chlo-
rination.In fact, chlorinationof this type of bulking sludge
can result in severefoaming problems.
Chlorinationof the activatedsludgein SBRs for fila-
ment control poses special problems becausethe aeration
basin is not mixed for the entire SBR cycle. To obtain the
proper dosing conditions and dose level, chlorine should
only be added to the activated sludge when it is being
aerated.One approachfor controlling the chlorine dose
level and addition conditions is to temporarily install an
RAS systemin which the activatedsludgeis pumped out
of the SBR, chlorinated in a hose or a tank, and then
returnedto the SBR.
Four case histories are presentedto illustrate the use
of chlorination for bulking control. These include two
from municipal wastewater treatment plants illustrating
the successfullong-termuseof RAS chlorinationand two
from industrial wastewatertreatmentplants illustrating the
use of chlorine added directly into the aeration basin.
b. City of Albany, CA
At the time these data were collected, the City of Albany,
GA, was a 20-MGD (0.88 m3ls,design) activatedsludge
plant receiving 13 to 16 MGD (0.57 to 0.69 m3/s) of
wastewaterin which approximately 5OVoof the BOD, and
Control of ActivatedSludgeBulkingand Other SettlingProblems
SS loads were from industry (paper processing and con-
venience foods manufacturing). The plant experienced
bulking problems to such a degreethat with only approx-
imately 12 MGD (0.52 m3/s) of influent flow, attainment
of secondaryeffluent criteria (30 mg/L monthly average
for BOD, and SS) was not possible.
The permanentRAS chlorination systeminjected chlo-
rine solution into a series of well mixed RAS wet wells.
The installation consistedof a Wallace and Tiernan 2200
lb/d (1000 kg/d) capacity chlorinator and a 2-in. (50 mm)
injector with 2 in. (50 mm) PVC injector water and chlorine
solution lines. The chlorine injection systemwas enclosed
in sheetmetal for protection.The chlorine solution line was
a 2-ir. (50-mm) PVC line leading to a 16 ft (5 m) long,
2-in. (50-mm) PVC headerwith 0.25 to 0.625in. (10 to 15
95
FICURE 4.14 Phasecontrast micrographsshowing pro-
gressive effects of chlorination on Thiothrix I: (a) healthy
organism, (b) loss of sulfur granules, (c) 50Vodamage, (d)
907o damage, and (e) over-chlorinated sample. (Original
magnification 1000x.)
mm) diameter holes at 4 rn. (100 mm) centers. The system
was constructedin November 1977 for under $5000.
Figure 4.15 presents6 y of data on RAS chlorination
atAlbany, GA. When chlorination for bulking control was
initiated, the RAS chlorination system did not exist.
Becauseof the urgency to control SVI, chlorine solution
was added to a wet well where the entire RAS stream
mixed with primary effluent. This mode of chlorination
startedon Week 15 and continued until Week 26 when the
change to RAS chlorination was made. While chlorine
solution was added to the mixture of RAS and primary
effluent,dosesof 5 to 15 kg Clll03kg SS/d were needed
to control bulking. When the chlorine dose point was
changed to the RAS wet wells, doses that satisfactorily
controlled bulking for the RAS/primary effluent mixture
 96 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationproblems

-S H ? :o o
=E6200
.E 100
. He e - 15
EEEA 5
? 3F= 7 0
& Ee

;*
93
so
E n :o
l0
J
! >^
EH ro
tB lo
0
l0 30 50 70 90 110 130 150 170 190 210 230 250 270 290
Weeknumber

FIGURE 4.15 Control of bulking by RAS chlorination at the Albany, GA wastewater treatment plant. (From Chartered Institution
of Water Environmentand Management,London. With permission.)

were "overdoses"for the RAS alone. Figure 4.15 shows h

that, even though the chlorine dose declined during the
}F ts
changeof dosepoint (from 4.7 to 2.9 kg Clrll03kg SS/d), =4 9 3
*gx 2 .v\/\'-
secondaryeffluentSS resultingfrom floc break-upcaused o t-
d'.ts .9 I
by the chlorine overdoseincreased. = -=
aE e 0
At various times during the period covered by oo
Figure4.l5, the target SVI was changed and in some
instancesRAS chlorinationwas not initiated soonenough 5.0
to prevent secondaryeffluent deterioration(e.g., weeks oQ( n
6<? 2.5
180 through 200). To illustrarethe chlorine dosing tech-
suz 0
nique for bulking control as employed by the City of uoo
Albany, a specificbulking incident is examinedin detail
in Figure4.16.The targetSVI was 230 ml/g. The chlorine 300
dosewas startedat low levels and gradually increaseduntil .7""svl=131m1/c
the SVI respondedby stabilizing and then falling. In the
-ao 200
middle of this period (May 30 through June 6), the SVI
fell below the target value. The RAS chlorine dose was
reducedin response.From June 18 through 20, the SVI
droppedto approximately100 to 130 ml/g. The chlorine
dose to the RAS was reduced and then turned off.
i
7) 100
Tu\*[
l ll 2t 31 t0 20 30
c. City of San Jose/Santa Clara Water Pollution
May 1979 Jun1979
Control Plant, CA
At the time that these data were collected, the plant pro- FIGURE 4.16 Use of targetSVI to control RAS chlorinationdose
for bulking control at the Albany, GA wastewater treatment plant.
vided tertiary treatment to 100 MGD (4.4 m3/s)of munic-
(From Jenkins, D. et al. (1982), in Bulking of Activated Sludge:
ipal wastewater.From July through September(peak load
Preventative and Remedial Methods, Chambers,B. and Tomlinson,
season), wastewater flow increased approximately 20Vo
E.T., Eds., Ellis Horwood, Chichester.With permission.)
and BOD, loading approximately doubled due to cannery
wastewater discharges. Effluent discharge criteria The plant had two stages of activated sludge with
included 30-d average BOD, and SS concentrations of tertiary effluent filtration. Plant upsets due to bulking in
l0 mg/L each and receiving water undissociatedammonia the secondary activated sludge system occurred during the
levels that dictated virtuallv complete nitrification. peak load seasons of 1979 and 1980. RAS chlorination in
 Control of ActivatedSludgeBulkingand Other SettlingProblems 97

solution into the secondary and tertiary activated sludge

system RAS pumps for over 15 y produced no deleterious
140
bo
effects on thesepumps, even though the secondarysystem
) l2o RAS pumps had brass bearings and cast iron impellers.
E
tnn Great care was taken to make certain that chlorine solution
-i
Ch
was never fed to an out-of-service pump.
80
RAS chlorination for control of filamentous bulking
60
€ fed to Mixed liouor channel was developedto a high degree(Figure 4.18). Chlorine
U; Totaluse= 8200Ks fed to
zg 12 doses were adjusted using target SVI values and micro-
RAS pump
=
use 2100 scopic observation of the mixed liquor. During the peak
i{ 8
9J
-d
load seasonof 1981, extremely low target SVI values
Q4
il (60 to 80 ml/g) were used in the secondary activated
sludge system so that high SS loading rates (30 to 50 lb
0 5 r0 t5 20 25 SS/fPld or 150 to 240k9 SS/m2ld)could be applied to the
Date,Mar1981
secondary clarifiers. To maintain these low SVI values,
FIGURE4.17 Comparison of RAS chlorinationeffectiveness very high chlorine doseswere often used in the secondary
when dosing chlorine to the mixed liquor channeland RAS activated sludge system (8 to 16 kg C1rl103kgVSS/d;
pump.(FromBeebe,R.D.et al. (1982),paperpresented at 55th Figure 4.18a).Dosesof this magnitudecausedsignificant
Annual Conferenceof the WaterPollutionControl Federation, floc destruction,resultedin turbid secondaryeffluents,and
St. Louis,MO. With permission.) caused an increase of secondary effluent dissolved total
L- both the secondaryand tertiary activated sludge systems
organic carbon (TOC) concentration from approximately
15 to 35 mg/L. The elevated turbidity and TOC values
and supplementaloxygen and ammonia were usedto com- could be tolerated at the plant because the downstream
L bat bulking during the 1981 and 1982 peak load seasons tertiary activated sludge system polished the effluent.
(Beebe et al., 1982). Maintaining a high-quality secondaryeffluentwhile using
such high chlorine doseswould be difficult in a single-
In the secondary activated sludge system, the initial
stage activated sludge plant.
chlorine injection point to the RAS was through a PVC
pipe with a 4-ft (1 .2-m) freefall into a 20 ft x 16 ft x 13 ft RAS chlorination was also effective for bulking con-
(6 m x 5 m x 4 m) RAS wet well. It soonbecameevident trol in the tertiary activatedsludgesystem(Figure 4.18b).
that this rurangementwas ineffective and chlorine could At the chlorine doses used during the 1981 peak load
season (2to 4 kg Clrll03kgVSS/d; 1.5-3.0mg Cl"/L dose
be smelledin the vicinity. Extensionof the 2-in. (50-mm)
PVC pipe to a point 5 ft (1.5 m) below the RAS surface concentration in the RAS stream), nitrification efficiency
was unaffected. Following initiation of RAS chlorination,
in this wet well virtually eliminated the loss of gaseous
the SVI decreased much faster than in the secondaryacti-
chlorine, but SVI control was still difficult. Doses of 8 to
vatedsludgesystem.Whetherthis was due to the presence
10 kg Clll03kg SS/d had little effect on SVI and created
of the more germicidal free chlorine in the tertiary system,
a turbid secondaryeffluent.
to differencesin the types of filamentousorganismscaus-
Following these poor experiences,two types of chlo-
ing bulking (Type 0041 in the tertiary system and Type
rination devices were installed. Diffusers were located
1701in the secondarysystem),or to differencesin sludge
about mid-depth across the full widths of aerated mixed growth rates is not known.
liquor channels leading from the aeration basins to the
secondaryclarifiers. Single outlet injectors were installed d. Stroh Brewing Co., Longview, TX
through the clean-outports about 6 in. (15 cm) upstream At the time these data were collected, Stroh operated a
of the closed-impeller, low-speed centrifugal RAS pumps brewery and container manufacturingfacility in Longview
(Figure4.11b). (Campbell et al., 1985).The combined wastewaterflows
The data presentedin Figure 4.17 show that the RAS from these plants were treated by an extended aeration
pump chlorine solution injection point, with its superior activatedsludgeplant; total wastewaterflow was 1.75 MGD
initial mixing, provided the more rapid, predictable and (0.08 m3/s). Influent wastewater characteristics were
efficient SVI control. Approximately 20 times more chlo- (mg/L) BOD5 - 1,500,COD = 2,500,TSS = 800, TKN =
rine was neededto control SVI when chlorine was dosed 7, and total P = 25. The treatment facilities consisted of
to the RAS well rather than to the RAS pumps. A similar a mechanically-cleanedbar screen,grit removal, ammonia
chlorine solution injection system was installed in the addition, two parallel completely mixed aeration basins
tertiary activated sludge system and it controlled bulking and secondaryclarifiers, a chlorine contact chamber, and
without compromising nitrification. Injection of chlorine post-aeration.
9B Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

100
I4 (a) First stagesystem . "Chlorine" dose
t2
10 o

8 a a aa
aa

6 aaa a a a a oa aa 40
E
a 4 aa
a oa

2 aaa aaa aaaaaaaa @

d

0
U 14 (b) Second stagesystem
F:

bo dU o
d l2
10
U 8
40
o
4
20
2
0
1 Ju l 3l Jul 10Aug 20 Aug 30 Aug 9 Sep 19Sep
Date,1981

FICURE 4.18 SVI and chlorine dose to RAS at the San Jose/SantaClara water pollution control plant during the 1981 peak load
season:(a) first stageactivatedsludge and (b) secondstageactivatedsludge. (From Beebe,R.D. et al. (1982), paper presentedat
55th Annual Conferenceof the Water Pollution Control Federation,St. Louis, MO. With permission.)

The Longview plant had always been subjectto fila-
mentous bulking problems that were most severefrom
April through October. The causative filamentous organ-
isms usually were Types 1701and 021N. The poor settle-
ability caused by large amounts of these filamentous
organismsrequired the addition of significant quantities
of cationic polymer to produceacceptablesludgesepara-
tion. In an effort to improvesludgesettleabilityand reduce
polymer costs, chlorination was practiced during 1979.
Chlorine solution was fed to the RAS at a rate of 3 kg
C121103 kg MLSS/d - the maximum rate that could be
achieved with the existing chlorine dosing equipment.
Throughout the 19-wk period that chlorine was dosedto
the RAS, filamentousorganismgrowth was largely unaf-
fectedand sludgebulking was not controlled(Figure4.19)
becausethe sludge inventory was not exposedfrequently
enoughto chlorine. With an aerationbasin HRT of approx-
imately 4 d, the activated sludge inventory passed the
chlorine dose point on the RAS line approximately
0.4 times/d.
By 1982, the single chlorine solution dose point on
the RAS line was replaced by two dose points in each
aeration basin, each located close to a surfaceaerator and
6 ft (2 m) below the liquid surface.Chlorine solution was
added at an overall mass dose rate of 3 kg ClrllO3kg
MLSS/d, and was split evenlyamongthe four dosepoints.
Figure 4.19 showsthe resultsof using this in-basin chlo-
rination system for bulking control from 1982 through
1984. With approximately the same organic loading, the
total costs for bulking control chemicals (polymer plus
chlorine) were reduced from $66,600/y in 1979 to
$8,800/y in 1984 with the in-basin chlorination system.
(Table4.2\.
e. Plastics Manufacturing Wastewater Activated
Sludge System, WV
This exampleillustratesthe effectsof a chlorine overdose
on activatedsludge and demonstratessystem recovery for
a completely mixed activatedsludgeplant treating 1.4 to
3.0 MGD (0.06 to 0.13 m3/s) of plastics manufacturing
wastewater(Campbell et al., 1988) Typical wastewater
characteristicswere (mglL), COD = 1200 to 1800,
BOD, = 700 to I100, and TSS = 30 to 100. Typical F/M
values were 0.2 to 0.5 kg COD/kg MLSS/d with MLSS
levels between 3000 and 5000 mg/L.
Activated sludge settleability deteriorated as the
MLS S concentrati on i ncreased. W hen S V I s wer e
>120 mL/g, secondaryclarifier sludge blanket levels and
effluent SS concentrationsincreasedand sludgedewatered
poorly in the centrifuges, reducing the solids removal
capacity of the system.When this happened,the MLSS
concentrati ons i ncreased and caused further SVI
increases,and so on.
The major filamentous organisms causing bulking
were Type 1702 and, to a lesser extent, N. limicola II.
Type 1702 grew largely inside the flocs, making them
diffuse. Type l7O2 is related to Type 1701 (Eikelboom,
1977), the growth of which in activated sludge is caused
by a DO concentrationtoo low for the applied F/M (Palm
et al.. 1980).
Control of ActivatedSludgeBulkingand Other SettlingProblems 99

400
300
t979
1\
,^.nffi
200
Cl, fed to RAS
100

oo 400
t982
E 300
\Z\
X
o 200 N
q
100 \/a C12fed to
aeration basin

400
o 1983
300
(h 200
100 Cl2 fed to
aeration basin

400
300 t984
200
100 Cl2 fed to
aeration basin
Jan Mar MaY SeP Nov
ri... ruJ.ll,

FIGURE 4.19 Weekly averageSVI from 1979to 1984 atthe Stroh Brewing Co. activatedsludgeplant, Longview, TX. (From Campbell,
H.J., Jr. et al. (1985), Proceedingsof 40th IndustrialWaste Conference,Purdue University, West Lafayette, IN. With permission.)

TABLE4.2
Annual Usageand Cost of BulkingControl Chemicalsat the Stroh BrewingCo. Activated
SludgePlant, Longview,TX
Year 1979 1980 1981 1982 1983 1984

AverageBOD, load, 103lb/d 22,000 20,000 23,000 23,000 20,000 23,000

Chlorinedosepoint RAS Aeration Basin Aeration Basin Aeration Basin
Polymerconsumption,gal/y 7600 2160 2360 1090 213 182
Chlorineconsumption,103lb/y 30 00432761
Polymercost,$/y 63,000 18,000 19,700 9100 1800 1500
Chlorinecost,$/y 3600 0 0 5200 3200 7300
Totalcost,$/y 66,000 18,000 19,700 14,300 s000 8800
AverageSVI, mllg 285 286 t64 160 159 156
Source From Campbell, H.J., Jr. et al. (1985), Proc, 40th Industr. Waste Conf., Purdte University, West Lafayette, IN,
p. 759. With permission.

An in-basin chlorination system that delivered chlo-
rine solution through four dosepoints in the aerationbasin
was first testedon March 9.1984. when the SVI exceeded
I2O mL/g and activated sludge settling was significantly
impaired. Figure 4.20 depicts the chlorine addition rates
and SVI values throughout the test period. Thirty-minute
settleable solids were measured every 4 h and SVI was
calculated 2 times/d. An initial chlorine dose rate of 3 kg
ClzlIO3kg MLSS/d was applied for 3 d with no effect on
the SVI; in fact, the SVI continued to rise. On March 12,
the chlorine dose rate was increased to 5 kg Clrl103 kg
MLSS/d and held at this level for about 5 d. Again, no
noticeable improvement in sludge settleability occurred.
On March 16, 1984, the chlorine dose rate was
increasedto 6.7 kg Clzll}3 kg MLSS/d and after approx-
imately 3 d at this rate, the SVI started to decrease.On
March 19, the chlorine dose rate was reduced to and
remained at 4 kg Cl2/103kg MLSS/d for 3 d. The SVI
continued to decreaseslowly to approximately I20 mL/g.
On March 22,the chlorine doserate was increasedto 8 ke
100 lr4anualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

160 l4

r50 High secondary cltrifier

t2 dudse blanket level t2
140
120
r30 to6
z
J 100
t20
)E :. U;
o80 o
llo U

tr 6d
100
460

=
a U

'70
2

60
l8 t4
tlo,". APr MtrDot.. APr
198,+
'nro

FIGURE4.20 SVI andchlorinedoseat a wastewater activated
sludgeplantat a plasticsfactoryin March1984.(FromCampbell,
H.J.,Jr. et al. (1985),Proceedingsof 40thIndustrialWasteCon-
ference,PurdueUniversity,WestLafayette,IN. With permission.)
Cl2l103kg MLSS/d. After a little over 3 d at this chlorine
dose rate, the SVI decreasedrapidly to between 70 and
80 ml-/g. The chlorine dose rate was reduced to 5 kg
C12/103kg/d for 2 d and stoppedon March 27, 1984. The
SVI increasedto about 90 ml-/g and leveled off.
FIGURE 4.21 Effluent TSS concentrationand chlorine dose at
a wastewateractivatedsludge plant at a plastics factory in March
1984. (From Campbell, H.J., Jr. et al. (1985), Proceedingsof
40th Industrial Waste Conference, Purdue University, West
Lafayette, IN. With permission.)
l4
t2
tz o
! to3

Microscopic examinationof the activatedsludgedur- i ttxr

ing in-basin chlorination showed that SVI reduction by O
* 80 U
chlorine was accompaniedby significantfilamentand floc 3,
damage.As the chlorine dose was increased,Type 1702 960
filaments successively showed deformed cells, empty
spaceswhere cells had lysed, completely empty sheaths, U

and broken sheaths.Flocs showed progressivelygreater

break-upwith productionof fine suspendedparticlesindi-
cating a chlorine overdose. High chlorine doses were t8

required because the Type 1702 filaments grew largely

within the floc and significant floc destruction was
requiredfor the chlorine to get at the Type 1702 filament.
These results demonstratethat in-basin chlorination
at high chlorine doses can adversely affect effluent TSS
(Figure 4.21), soluble COD (Figure 4.22), and turbidity
levels. For example, following the use of an overall chlo-
rine mass dose of 8 kg Clrl103kg SS/d, the effluent TSS
concentration increased to 60 to 80 mgll. from 30 to
50 mg/L and effluent soluble COD increased from 40 to
80 mg/L to as high as 150 mg/L. Theseresultsdemonstrate
the need for very careful control of the chlorine dose rate
and duration when practicing bulking control with in-
basin chlorination. They also illustrate that activated
sludge recovers quite rapidly from a chlorine overdose.
Effluent sampleswere analyzed for chlorinated organics
to determinewhether suchcompoundswere formed during
chlorine addition. With the exception of 1-2-dichloroet-
hene (DCE), other chlorinated organic priority pollutants
were presentat concentrationsnear or below their analyt-
FIGURE4.22 EffluentsolubleCOD concentrations andchlo-
rine doseat a wastewateractivatedsludgeplant at a plastics
factoryin March 1984.(FromCampbell,H.J.,Jr. et al. (1985),
Proceedings of 40th Industrial WasteConference,
PurdueUni-
versity,WestLafayette,IN. With permission.)
ical detection limits. Because DCE levels were below
100 pgll. and within the normal variability of routine plant
operation, it was concluded that the generation of chlori-
nated organic compounds during in-basin chlorine addi-
tion for bulking control was insignificant at this plant.
Similar conclusions were made by van Leeuwen and
van Rossum (1990) when using in-basin chlorination in
laboratory-scale experiments at the Daspoort, Pretoria,
South Africa activated sludge plant. No halo-organics
were detected in the secondary effluent when in-basin
chlorination at a rate of 8 kg Cl2/lO3 kg MLSS/d was
applied in the full-scale nutrient removal activated sludge
plant.
 Control of ActivatedSludgeBulkingand Other SettlingProblems 101

TABLE4.3
Use of HydrogenPeroxidefor Filamentous
BulkingControl
EffectiveHrO,
Concentration,
Location mg HrOr/L Reference

Petaluma,CA, full scale 9-68, average FMCCorp.(1973);

31.5 Caropreso et al.
( t 97 4 )
St. Augustine,FL, full scale 12 (basisunclear) FMC Corp. (1976)
San Jose,CA, lab scale,fill- 40-200 StrunkandShapiro
and-draw (1976)
Princeton,NJ, small full l00b Caropresoet al.
scale (r974)
Wilmington, DE, lab scale 40-200b Co le et al . (1973)
Washington,D.C., pilot 2040b Co le et al . (1973)
6.5 mm PVC valves
Textile mill, full scale 60 (basisunclear) Keller and Cole
(1973)
_,ri' i f\
o Based on wastewater inflow. /l\
b Basedon volume ofaeration basinand secondaryclarifier (singledose).
aeratlontank

4. Use of Hydrogen Peroxidefor Bulking Control

9.5 mm O.D . stai nl es s
a. General steeltubi ng
Two55galpolyethylene
lined
Hydrogenperoxide(HrOz)hasbeenusedfor bulking con- drums g 509o
oontainin H2O1-
trol in a similar fashionto chlorine.HrO, dosesand appli-
cation times for effectivefilamentousorganismreduction FIGURE4.23 Feed systemfor dosinghydrogenperoxide.

t_ and bulking control vary from plant to plant (Table 4.3)

but generallyare higher than for chlorine.
(FromFMC Corporation, With permission.)
Philadelphia.

Keller and Cole (1973) statethat the time requiredto
reduce the SVI to 50Vo of its initial value (actual SVI
valueswere not given) was a function of HrO, dose.At a
dose of 0.4 kg HrOr/kg MLSS/d, a 50VoSVI reduction
occuned in lessthan I d while at a doseof 0.1 kg HrOrlkg
MLSS/d, a 50% SVI reduction took place in 8 d. Using
these data, Keller and Cole (1973) concluded that the
minimum effective HrO, dose for bulking control was
approximately0.1 kg HrOr/kg MLSS/d. In experiments
confirming this value,SVI was controlledusing a doseof
0.15 kg HrOrkg MLSS/d but not with a doseof 0.035 kg
HrOrkg MLSS/d.
Both batch and continuousdosing of HrO2 have been
used to control SVI. Dose points locatedin the aeration
basin, the RAS line, and the mixed liquor channel have
been used. Like chlorine, excellent initial mixing of HrO2
and activated sludge is required for efficient filamentous
organismdestruction.Data from St. Augustine,FL (FMC
Corporation, 1976) suggestedthat the contact time
betweendosedHrO, and activatedsludge should be longer
than for chlorine. A dose of 20 mg H2O2/L to a degritter
located 5 min flow time upstream of a secondaryclarifier
caused foaming but did not control bulking. When the
HrO, dosepoint was relocatedto a point 15 min flow time
upstream of the secondaryclarifier, a dose of 12 mg
H2OzlLreducedSVI from 580 mllg to 178 ml-/g in 2 d.
The HrO, solutionusedfor dosingis usually507ovlv.The
recommendedfeed systemis illustratedin Figure 4.23.
A comparativestudy of the effectivenessof chlorine
and HrO, for bulking control was conductedat an acti-
vatedsludgeplant treatingthe wastewaterfrom a recycled
paper mill. Both chemicalswere able to control filamen-
tous bulking due to Types0675, 1851,and 0041, but the
HrO, dose required was five times the chlorine dose
required. Since the cost of HrO, was twice that of the
chlorine, the total bulking control cost of the HrO, was
ten times that of the chlorine.
Caropresoet al. (1974) statethat HrO, destroysfila-
mentousorganismsby attackingtheir sheaths.Regardless
of mechanism,the effect observedis similar to that when
chlorine is used, i.e., the filamentsbreak up and become
shorterand cells within the filamentsshow signs of lysis.
In our experience,the doseresponseof filamentsto H,O2
appearsto be all or nothing rather than an increasingeffect
with increasingdose,as for chlorine.
Caropresoet al. (1914) also claim that, in addition to
killing filamentousorganisms,HrO, producesoxygenthat
may supplementDO:
102 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

300
800

700 Control sludge mean

200
@

600 E
Fi

(h

Ozonated sludge mean

500
d

; 400 Start H2O2

ct treatment

d
Periodof oPeration,

FICURE 4.25 Effect of ozonationon activatedsludgesettling.

(From van Leeuwen,J. and Pretorius,W.A. (1988),J. Inst.Water
Environ. M gt., 2, 223. Wrrh permission.)
End H2O2
treatment 5. Use of Ozone for Bulking Control
100

Pilot-scaleexperimentson the addition of ozone (O.) to

'7 the aerobic stage of a three-stage(anaerobic,anoxic, aer-
t'7 24 31
Jul Aus obic) BNR activated sludge plant was conducted by van
Date,19'74 Leuwen and Pretorius(1988) and van Leuwen (1988).
Ozone dosesof 3, 6, and 9 mg/l- (basedon wastewater
FIGURE 4.24 SVI responseto hydrogenperoxide treatmentat flow) and equivalentto 2, 4 and 6 kg 03/103kg MLSS/d
the Petaluma water pollution control plant in California. (From were used. Activated sludge settling was not affected at a
Caropreso, F.E. et al. (1974), Bull. Calif. Water Pollut. Control dose of 2 kg Or/103kgMLSS/d. Doses of 6 and 9 kg
Assn.,p. 44. With permission.)
0./103kg MLSS/d were equallyeffectivein reducingaver-
age SVI from approximately 200 mL/g to approximately
125 mL/g (Figure 4.25).
ZH.O., -+ 2HrO + O,
In addition to providing SVI control, an O, dose of
6 kg Or/103kg MLSS/d decreasedeffluent color, turbidity,
Should bulking be causedby low DO, this oxygen
and TSS concentrations.Ammonia removal by nitrification
could help amelioratethe problem. Conversely,if the acti-
improved somewhat and enhancedbiological P removal
vated sludgebacteriadevelopthe ability to rapidly degrade
was virtually unaffected. Halo-organics were not formed
the HrO, before it becomes available for killing filamen- during ozonation and the halo-organicformation potential
tous organisms (through production of peroxidases), its of the ozonatedactivatedsludge effiuent was less than for
bulking control effectivenessmay be compromised. the unozonated activated sludge effluent (van Leuwen,
b. Citv of Petaluma, CA 1988).

During 1974, the City of Petalumawater pollution control

plant that treated mixed domestic and industrial wastewa-
ters experiencedseverebulking problems with SVI values
in the range of400 to 700 rnlJg. Hydrogen peroxide (as a
507ovolume HrO, solution) was dosedto the final quadrant
of a two-passaerationbasin that was fed primary effluent
atthe 1/rpoints and RAS at its head end (Caropresoet al.,
1974). Figure 4.24 shows the dosesand concentrationsof
HrO, used over a 4-d period and their effect on SVL The
HrO, doses ranged from 9 to 68 mg H2O2|I' and were
variously applied for 8 to 24 hJd.During this bulking inci-
dent, 2300 lb (1050 kg) HrOr (1007obasis) was used to
bring the SVI under confrol (from 550 mL/gto 300 ml/g).
The SVI continued to decreaseafter HrO, dosing stopped.
6. Filamenticides
In recent years, severalso-calledfilamenticides have been
marketed.Our experiencewith thesematerials is that they
appear to kill filaments, but their toxic properties persist
through activated sludge processing and produce effluent
toxicity.
METHODS
IV. SPECIFIC
OF BULKINCCONTROL
This section discussesspecific methodsof bulking control.
These methods can constitute the second phase of an
 Cont r olof A c t i v a te dS l u d g eB u l k i n ga n d Oth er S ettl i ngP robl ems
attack on a bulking problem - the first phase is the use
of the previously described, rapid, nonspecific chemical
methods. Specific bulking control methods can also be
used in designingnew plants.To use specificmethods,it
is necessaryto characterize the causative filamentous
organisms.
The use of filamentous organism characterization as
a diagnostic tool for the causesof activatedsludge bulking
was discussedearlier.In this section,the relationshipof
filamentousorganism growth conditions (where known)
and methodsfor bulking control will be discussed.
A. Nurnrrrur
Drncrrrucv
1. General
Activated sludge nutrient requirementsfall into two cate-
gories based on the amount of nutrient required to form
biomass: macronutrients(N and P) and micronutrients
(Ca, Mg, Fe, etc.).Nutrient deficiencyin activatedsludge
plants treating food processingor pulp and paper waste-
water or a combination of thesewastewatersand municipal
wastewatersis generallycausedby insufficientN andTor
P becausethe carrier (potable water) for thesewastewaters
usually contains sufficient micronutrients.Wastewaters
from chemicalprocessingand other industriescan be defi-
cient in both macronutrientsand micronutrientsbecause
chemical processesmay remove nutrients from the carrier
water (for example,by precipitation) or becausethe carrier
t_ water may have been demineralizedleading to removal of
both macronutrientsand micronutrients.
Nutrient deficiencyis diagnosedby a combinationof
wastewateranalysisand microscopicexaminationof the
activatedsludge.If a microscopicexaminationrevealsany
of the following, a macronutrient deficiency could be
occurring:
. for both cell growth and maintenance(endogenousrespi-
Major filamentsare Type 02lN,Thiothrrr spp.,
S. natans, H. hydrossis, and N. limicola lll.
(Note that these filamentous organisms have
causesother than nutrient deficiency.)
. Viscous activated sludge showing significant
amounts of exocellular material by India ink
reverse staining
. Aeration basin and/or secondaryclarifier foam
(scum) containing significant amountsof exo-
cellular material (often Neisser-positive) but
not c on ta i n i n g n o c a rd i o fo rm o rg ani sms,
M. parvicella, or Type 1863
Poor activated sludge settling due to viscous activated
sludge causedby nutrient deficiency cannot be controlled
satisfactorily by chlorination or HrO, addition. Sludge can
be made to settle with difficulty by large dosesof polymer.
With P deficiency in particular, the danger is that, if the
I
103
nutrient deficiency becomes more severe, the ability to
remove soluble organic matter will be lost and treatment
will fail.
2. Macronutrient Deficiencv
a. Ceneral
When a microscopicexaminationsuggestsnutrient defi-
ciency and macronutrientdeficiency is the likely cause,
the BOD', (COD, TOC) to N and P ratios of the waste-
water influent to the aeration basin and the total P and
TKN content of the mixed liquor VSS should be deter-
mined. It is commonly stated (e.g., Metcalf and Eddy,
2003) that sufficientmacronutrientsare presentwhen the
wastewaterfeed to the aerationbasin containsBOD.:N:P
in a weight ratio 100:5:1.
b. FactorsAffecting Macronutrient Requirements
The BODr:N:Pratioof I 00:5:I is basedon the assumption
that the net sludge yield is 0.5 gVSS/g BOD. removed
and that the sludge contains l07o N and 27o P on a VSS
basis.Macronutrientadditions of less than this ratio are
used in some industries.For example,Richard and Cum-
mins (1997) showedno signsof P deliciency(including
no increasein sludge polysaccharidecontent) in an acti-
vated sludgeplant treatingpapermill wastewaterat inffu-
ent BOD./P ratios of 100:0.2to 100:0.4.Howeversome
other pulp and paper wastewatersproducedP deficiency
in activatedsludgeswhen the influent BOD./P ratio was
less than 100:0.5.The specificmacronutrientneedsof an
activatedsludgeare influencedby sludgeage and temper-
ature. Significant recycling of macronutrientsoccurs in
high MCRT activatedsludge from cell lysis so that less
macronutrientis neededto treat a given amountof BOD.
than in lower MCRT systems.For a given system,macro-
nutrientrequirementsare higher at low temperaturesthan
at high temperatures.Influent BOD. is used by bacteria
ration). Cell growth on influent BOD. requires N and P
but cell maintenanceusesBOD, without using externally
suppliednutrients.More of the influent BOD, is usedfor
cell maintenanceat high temperaturesthan at low temper-
aturesso that the macronutrientrequirementsto remove
BOD, are lower at high temperatures.Sludge production
(cell yield) increasesat low temperaturesfor the same
reasons.
c. Availability of Macronutrients
The availability of N and P in the aerationbasin influent
should be determined.lf the wastewatercontainsreadily
metabolizable carbon sources (e.g., simple sugars or
organic acids) and the major N or P source is organically
bound, the macronutrients may not be readily available
enough for use during the metabolism of the carbon
source.An example of this was encounteredduring the
summer at the San Jose plant when it received a mixture
104
M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
of municipal and fruit and vegetablecanning wastewaters.
When determining the need for N supplementationfrom
wastewateranalysis,only the primary influent NHr-N was
counted even though primary influent also contained
organic N. It was felt that the organic N would be available
too slowly to keep pace with the rate of carbon substrate
metabolism. Further, becausethe sewer system was long
and the sewage temperatureswere high, it is likely that
some of the influent organic N had already been incorpo-
rated into biomass grown in the sewer system prior to
reaching the plant.
Both NH.-N and NO,-N are readily available N
sourcesfor activatedsludgegrowth. In nitrifying systems,
NO'.-N is available as an N source unless it is removed
by denitrification(to Nt) in anoxic conditions.
Nitrogen oversupply can be problematic in nitrifying
activatedsludge plants becausethe N in excessof require-
mentsis convertedto NOr. ShouldNOr-N levelsof >8 mg
N/L (approximately) be produced, denitrification may
result and causesludge flotation in the final clarifier. For
this reason,it is bestto measuretotal inorganicN residual
(NH1-N + NO'-N + NO.'-N) for regulating N addition
to N-deficient activatedsludge systems.Unless the plant
has an effluent requirement for P, an operator has no need
to worry aboutaddingit in excess(exceptfor costimpacts).
This problem can be especially seriousin industrial
wastewateractivatedsludgeplants that use a commercial
agricultural N fertilizer that contains a mixture of urea,
ammonia, and nitrate (usually 24 to 257o of the N is
NOj-N). Using this mixture causesproblemsbecausethe
NH3-N residualis usuallyvery small but the NO.,-N resid-
ual builds up to high values,leadingto denitrification.For
this reason,when using f'ertilizersfor N supplementation,
formulationsthat do not containnitratearerecommended.
Similar situationscan exist for P in industrialwastewater
treatmentplants.For example,when P bound organically
as phytin in a soybeanprocessingwastewatertreatment
plant was not availableat a high enough rate to supply
adequatesolublephosphate,Type 02lN bulking resulted.
The formation of sparingly soluble phosphateprecipitates
when the aerationbasin is operatedat about pH 9 may
limit P availability.Activated sludge plants treating pulp
and paper wastewaterfrom a processthat useslime must
dose P at a much higher rate to avoid P deficiencythan a
plant that doesnot use lime. When phosphateand a strong
basefor neutralization are addedto an influent, care should
be taken to preventlocally high pH and high P areaswhere
P precipitation may occur.
McKeon et al, (2001)found that the dischargeof water
treatment plant sludge containing iron to the collection
system serving the Philadelphia SoutheastWater Pollution
Control Plant reduced infl uent-dissolvedP concentrations
to <0.2 mg P/L and caused P deficiency in the activated
sludge system at the wastewatertreatmentplant. The defi-
ciency was relieved when the water treatmentplant started
Pr oblem s
using alum as the coagulant or when phosphoric acid was
added to the wastewatertreatment plant influent.
The Detroit WastewaterTreatmentPlant employs iron
salt addition for P removal to the raw wastewaterfollowed
by primary sedimentation and high purity oxygen acti-
vated sludge. P deficiency was observed in the activated
sludge system when secondary treatment influent-dis-
solved P concentration was <0.2 mgll- (Franklin et al.,
2001). Similar effects were observedat the Rock Creek
Advanced WastewaterTreatment Plant in Hillsboro, OR.
d. Satisfying Macronutrient Demands
Macronutrient supply should match macronutrientdemand
to the extent that macronutrientsare never depleted any-
where in the aeration basin. Plants treating macronutrient
deficient wastewaterswith highly variable organic loads
often oversupply macronutrients during periods of low
organic loading to provide reservesupplies that help meet
macronutrient demandsfrom high organic loads.
The importance of never running out of macronutri-
ents is illustrated by the pure culture chemostatexperi-
ments of Richard et al. (1985) on the NH.-N-limited
growth of Type 02lN and a floc former isolated from
activatedsludge.With a steady-stateNH3-N supply, the
growth rate of Type 021N was neverhigh enoughrelative
to the floc former to become dominant (Figure 4.26).
However,with an intermittentNH3-N supply,Type 02lN
took up NH3-N more rapidly than the floc former
(Figure 4.27). This allowed Type 02lN to dominate (and
bulking to occur) when the NHI-N supply was intermit-
tent and was at times depleted. Conversely,when the
2.0
! t\
o
l(l
'
rl
--- Floc former
_ Type 02lN
0.0
0123
NH1-N,pgN/L
FICURE 4.26 Monod model growth curves for ammonia-lim-
ited growth of Type 021N and a floc former isolated from acti-
vatedsludge.(From Richard,M.G. et al. (1985),J. WaterPollut.
Control Fed., 57, 1152.With permission.)
Control of ActivatedSludgeBulkingand Other SettlingProblems 105

NH3-N feed was continuous(no bulking). Type 02lN was

selected when J57o of the NH3-N was fed in a spike at
l--- I Floctbrmer 6-h intervals (bulking). See Figure 4.28. The task of add-
lt- t ing macronutrientsto a nutrient-deficient wastewaterwith
o-o 'a O- O T ype 02l N
a fluctuating organic load becomeseven more demanding
\ -r. when the treatment plant also has stringent dischargelim-
.l
Z tn
Ur
o
its for deficient macronutrients. This problem is being
faced for P by some industrial wastewatertreatmentplants
I
(e.g., pulp and paper mills in the Great Lakes region of
z r-r{..
F
I
North America). In such situations,it is not possible to
z -l*_ add excess P to take care of the increased P demands
\--t.. causedby high organic loads, becausethis could elevate
t' t the effluentP levelsbeyondthe concentrationlimits. This
problem has been addressedby (1) operating as closely
.
as possibleto the P requirementand (2) adding P precip-
itation and solids separation processesto the activated
20 sludge system. Both approachesare unattractive - the
34 former becauseof the risk of encountering P deficiency
Time,h and the latter becauseof its additionalcost.
Harperand Jenkins(2001) suggestedthat the situation
FIGURE 4.27 Batchcultureammoniauptakeof Type 021N and
an activatedsludge floc former previously grown under ammonia
could be handledusing an activatedsludgebasin with an
limitation and subjected to spikes in ammonia concentration. initial anaerobiczone performing enhancedbiological P
(From Richard,M.G. et al. (1985),J. WaterPollut. Control Fed., removal. In this type of process,soluble P is releasedin
5'7, 1152.With permission.) the anaerobiczone and taken up in the aerobic zone. P
would be added at a rate sufficient for the averageorganic
load. When organic load is low, the excessP would be
taken up aerobically,producing low effluent soluble P.
100 When the organic load is high, some of the P released
anaerobicallywould be usedfor growth. Again, the efflu-
ent soluble P would be low becauseof P uptake in the
.s 8o
aerobiczone.
'Eoo Harper and Jenkins (2001) validated this concept in
laboratory experiments.Completely aerobic (CA) and
u anaerobic-aerobic(AnA) systems(both P-limited) were
fed a constant amount of P and subjected to variable
' . 40
'j:
organicloading that includedtwo consecutivedays where
7
Bln the organic load was l07o of the average organic load.
s The AnA system had an average effluent soluble P of
0.3 mg Pll, and never exceeded0.6 mg P/L. The CA unit
0
0 24 68 10 t 214161820 averageeffluent soluble P was 0.9 mg P/L and on some
No. ofMCRTs days reached1.5 mg P/L.
Jobbagyet al. (2001) working with winery wastewater
FIGURE 4.28 Ammonia limited growth competition between that was severelyN- and P-deficient, showed that an AnA
Type 021N (o) and an activated sludge floc former (l) in chemo-
system produced an activated sludge that contained
stat culture with intermittent (I) and continuous (C) NH,-N feed-
approximately 2oo/oexopolysaccharideand 407o internal
ing. CA.l = 15 with intermittent feeding of 75Voof total ammonia
polysaccharidestorageproducts,while in a CA system,
at 6-hour intervals.(From Richard,M.G. et al. (1985),J. Water
Pollut. Control Fed.. 57. 1152.With oermission.) the activated sludge contained approximately 40Vo
exopolysaccharideand 207o intenal polysaccharide stor-
age products.This differencein activatedsludgecompo-
NH.-N supply was continuous and matched to the car- sition manifesteditself in differencesin settlingproperties.
bonaceous load so that it was never depleted, the floc The CA activatedsludgesettledvery poorly (SVI = 600 to
former dominated. 800 ml/g) becauseof viscousbulking causedby the very
Dual culture chemostat experiments with Type 02lN high exocellular polysaccharide content. The AnA acti-
under nitrogen limiting conditions (C:N ratio = 15:1) dem- vated sludge settledmuch better (SVI <250 ml/g) because
onstrated that the floc former was selected when the most of its polysaccharidecontent was intracellular.
106 M anua lo n C a u s e sa n d C o n tro lo f Ac t i vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
il20
d '"
d
80 90 r00 ll0
InfluentCOD/Pratio
FICURE 4.29 Effect of initial anaerobiczone on limiting influent COD/P ratio for a laboratoryscaleSBR activatedsludge(AnA =
anaerobic/aerobic;CA = completely aerobic). (From Harper, W.F. and Jenkins, D. (2001), Proc. Water Environ. Fed. WEFTEC
Conference,2001. With permission.)
Besidesproviding an ability to produce low effluent
P concentrationsfrom a P-limited activatedsludgewith a
fluctuating organic load, an AnA aeration basin also
appearsto have other advantagesfor P limited situations.
Because activated sludge developed in AnA activated
sludge contains higher amounts of glycogen and poly-
hydroxyalkanoate storageproducts that do not contain P,
the amountof P neededto treata given amountof organic
material is lower for an AnA system than it is for CA
activatedsludge.Harper and Jenkins(2001) showedthat
at a 4-d MCRT, CA activated sludge became P limited at
an influentCOD:P ratio of 100 to 110:1 while an AnA
system did not become P limited until the influent ratio
reached130 to 140:l (Figure 4.29).They were also able
to show that the lower P requirement was becauseof the
diversionof influent COD into internal microbial storage
productsthat did not require P for their synthesis.
e. Required Residual Macronutrient
Concentrations
Because each wastewater activated sludge combination
has its own unique macronutrient requirements,the suit-
ability of a specific BOD,:N ratio or BOD':P ratio in
meeting these requirements should be checked by mea-
surementsof effluent concentrationsof soluble (0.45 pm-
filtered) orthophosphate-P,NH.-N and NOr-N. The
required effluent levels will dependon the needsfor main-
taining N and P levels in the aeration basin as the influent
organic load fluctuates. However, concentrations of total
soluble inorganic N (NO3-N + NO'-N + NHr-N) of not
Pr oblem s
!
tr
tr
D
t20 130 140 150 160
less than 0.5 to 1.0 mg N/L and soluble P concentrations
of nof less than 0.1 to 0.3 mg P/L should be maintained.
The specific concentrationto be maintained can be estab-
lished by site-specificexperience.
Becauseit is possiblefor both N and P to be released
from activated sludge solids while they are present in the
secondaryclarifier, it is wise to check the soluble N and
P content of the mixed liquor as it leaves the aeration
basin.This analysisshouldbe performedon a samplethat
is filtered in the field immediately after taking to avoid
the releaseof N and P during transport to the laboratory.
Much higher effluent N and P levels may be needed
in somecases,especiallywhen high concentration,readily
degradable,soluble wastewaters(e.g., paper mill, food
processing, or brewery wastewaters)are treated in com-
pletely aerobic, well mixed aerationbasins. Here, mini-
mum N and P residuals of 1 to 3 mg/L may be required
to ensure sufficient nutrients in the floc interior to meet
the nutrient demand exerted by the high substrateuptake
rates associatedwith high concentrations of this type of
substrate.For example,Reid (1991) and Richard (1991)
showed that viscous and filamentous bulking (due to
N. limicola III) occurred in a Michigan paper mill acti-
vated sludge plant when effluent soluble P concentrations
were <1.0 mgP/l-. An effluent solubleP concentrationof
I to 2 mg P/L was neededto avoid these problems.
In some activated sludge effluents, especially those
from pulp and paper wastewatertreatment plants, the col-
orimetric analysis of P using Molybdenum Blue (e.g.,
stannous chloride and ascorbic acid reduction methods)
 Cont r olof A c t i v a te dSIu d g eBu l k i n ga n d O th er S ettl i ngP robl ems 107

(Water Environment Federation, 1998) can suffer from
positive interference.This can have serious consequences
if not detected because it can result in reduction of the
influent P feed rate based on the mistaken belief that
soluble P in the effluent is sufficient. On one occasion,
this resulted in P-deficient bulking that was detectedonly
when soluble orthophosphatewas detectedin the effluent
although no P feed was being added. An effluent sample
spiked with orthophosphateshould always be run to check
for P recovery. Brown colored lignin-derived materials in
paper mill wastewatertreatment plant effluents frequently
interfere positively with colorimetric tests for nitrate. A
NO.,- ion-specific probe is recommended for monitoring
effluent NO3-N conaentrationfor this condition.
3. Micronutrients
The principlesdescribedabovefor the diagnosisand cor-
rection of macronutrientdeficienciescan also be applied
to micronutrient deficiencies.The tasks of detectingand
resolving micronutrient problems are complicated by the
large numberof micronutrients;their (generally)low con-
centrations; small and often variable requirements; and
transitorydeficiencies.
The approachthat bestaccommodatesthesedifficulties
is to analyze the activated sludge solids for macronutrient
and micronutrientcontents.Table4.4 presentsinformation
on the N and micronutrient contents of the volatile matter
(VSS) in typical microbial biomass. Becauseactivated
sludgeVSS includes volatile matter other than microbial
biomass, for example, inert biomass and inert organic
solids, the micronutrient composition data in Table 4.4
cannot be used directly to assessthe micronutrient status
of the activated sludge.Rather a ratio of the concentration
of the micronutrients in VSS to the concentration of the
macronutrientN (TKN in VSS) is used.N is used as the
reference macronutrient becauseits levels in biomass are
Iess variable than P. It goes without saying that the first
step in this procedureis to ensurethat the activatedsludge
is not N deficient.The ratio of VSS micronutrientconcen-
trations to VSS TKN concentration is then compared to
the ratio in Table4.4. Micronutrientdeficiencyis indicated
if the experimentally determined ratio for a micronutrient
is significantlylower than the ratio indicatedin Table4.4.
Advantages of this approach are:
. The biomassis an integratorof the immediate
past history of the system (over the previous
one or two MCRTs). Thus, even if a micronu-
trient deficiency is only transitory,it will still
be detectablein the compositionof the biomass.
. Only one sample (the biomass) needs to be
analyzed.
. Some micronutrient levels in biomass are
higher than thosein the wastewaterso that ana-
lytical detectionand precision are improved if
the biomassmatrix does not presentoffsetting
analyticalproblems.
Micronutrient deficienciesare resolved in the same
fashionas macronutrientdeficiencies.The limiting micro-
nutrient is added to the activatedsludge system and its
responsein terms of treatmentefficiency,sludge quality,
and micronutrientcontentis monitored.Figure4.30 shows
the responseof an activatedsludgeplant treating a fface

TABLE4.4
TypicalBiomassNutrient Concentrations l 600 J
4

< 2) U t200
Nutrient Concentration,g/kg VSS Ratioto N
j 800
Nitrogen 125 .9 rnn
Phosphorus 25 i ro-' d
400
Potassium I4 l l x 10-' o

Calcium 14 1. 1x 1 0 - ' H rsn

o -"- U U
O
Magnesium l0 8 x 10-2 U -F
Sulfur 8.5 6.8x 10r F 100
Sodium 4.3 3.4x l0-2
Chloride 4.3 3. 4x 1 0 '?
=5n
Iron 2.8 2.2 x 1O-2 Cooling tower
Zinc 0.3 2. 4 x 1 0 3 t-- blowdown added
Manganese 0. 15 1. 2 x 1 0 3
Copper 0.03 2.4 x 104 123456789
Molybdenum 0.006 4. 8 x 1 0 5 Daysof operation
Cobalt <0.0006 4.8 x 10-6
FIG U RE 4.30 Effect of micronutrientadditionon COD removal
Source: Abstacted from Grady, C.P.L., Jr. et al. (1999),
efficiency at a peffochemical wastewater activated sludge treat-
Biological Wastewater Treatment, 2nd ed., Marcel Dekker,
ment plant. (From Sun, P.T. (2002), personal communication.
New York. 109.
With permission.)

I
108 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
element-deficient industrial wastewaterto the addition of
cooling tower blow-down. A dramatic improvement in
TOC removal efficiency occurred within days of the cool-
ins tower blow-down addition.
OxvcrN(DO)
B. Low Drssorvro
CorucrNrnnrrorus
1. G ener al
Low DO concentrationscan cause the growth of several
typesof filamentousorganismsin activatedsludge.At low
to moderateMCRTs, S. natans,Type 1701, andH. hydros-
sls are associatedwith low DO bulking. Type 1701 may
occur at more severeDO limitation than S. natans (Hao,
1982;Richardet a1.,1985).At high MCRTs, M. parvicella
growth is favoredby low DO (Slijkhuis, 1983).
Palm et al. (1980) conductedlaboratoryexperiments
on low DO bulking using settled municipal wastewater
feed and continuously fed, completely mixed aeration
basins. The DO concentrationrequired to prevent the
growth of S. natans was a function of the F/IVI.The higher
the F/I4, the greater was the DO concentration required.
These experiments establishedthe DO concentrations
requiredto preventlow DO bulking in a completelymixed
aeration basin over a range of FlN4 values. Figure 4.31
definesregions of Fltr{ and DO concentrationwhere bulk-
ing did and did not occur.A safe operatingline has been
drawn at +0.5 mg DO/L higher than the experimentalDO
concentrationthat resultedin nonbulkingsludge.A safety
factor was usedbecausethe SVI of the nonbulkingsludge
in some of the experimentswas at levels that would have
been unacceptablein practice.
1.8
h t.u
Bulking occurred
an
I Bulking did not occur
',
bol?
E
9 r.0
o 0.8
o other activatedsludge systems.TheseDO uptake rate data
o 0.6
O E
i "'"^ "
LL
0 can be determined for a given DO uptake rate.
0 1.0 2.0 3.0 4.0 5.0
AerationbasinDO, mg/L
FIGURE 4.31 Combinations of F/M and aeration basin dis-
solved oxygen concentrationsat which bulking and nonbulking
sludges appear in completely mixed, continuously fed aeration
basins.(From Palm, J.C. et al. (1980), J. WaterPolhu. Control
Fed., 52,2484. With permission.)
Pr oblem s
6
a xD O= 0.1-0.5mg/L x DO = 0.5 - 1.0mg/L
d
e o DO = 0.5 - 1.0mg/L e o DO = 0.5 - 1.0mg/L
E o.o
ftAl-q'"l
O n4
?, o.z
-il
E ooo
{J 400
1 zoo
at,
0 5 101520253035404550 55 6065
Dayof experiment
FIGURE4.32 Temporalvariationof F/M and SVI at aeration
basindissolvedoxygenconcentrationsof 0.1 to 0.5 mg/L and
0.5 to 1.0mg/L.(FromPalm,J.C.et al. (1980),J. WaterPollut.
ControlFed.,52, 2484.With permission.)
Palm et al. (1980) showedthat low DO bulking could
be causedand cured by manipulatingF/M and DO con-
centrations(Figure 4.32).The onsetof bulking was much
more rapid than its cure. Curing low DO bulking by
increasing aeration basin DO conceniration was achieved
by wash-out of the filamentous organism population,
which.requiredtwo or threeMCRTs. Thus for a plant with
a lO-d MCRT, 20 to 30 d would be requiredto reach the
lower steady-statefilamentousorganismlevel associated
with the higher DO concentration.This finding empha-
sizesthe importanceof having a rapid nonspecificmethod
suchaschlorinationfor lowering the filamentousorganism
population.
The safe operating line presentedin Figure 4.31 is
specific for the experimentsconducted by Palm et al.
(1980) and the results should be used only as guidelines
AA for other systems.Lau et al. (1984a and 1984b) showed
^^
lt)
.t)
that DO uptake rate was important in determining the
56
outcome of the competitive growth of S. natans and an
activatedsludgefloc-forming organism.Palm et al. (1980)
ta bo
developeda relationshipbetweenmixed liquor DO uptake
rate and F/M which allowed their data to be applied to
were used to construct the scaleon the right axis of Figure
10:3 4.31 so that the DO concentrationsrequired to control low
A-
DO bulking in a completely mixed activatedsludge system
6.0 The data presentedabove might seem to indicate that
the resolution of low DO bulking problems should be
fairly straightforward, i.e., one must either decreaseF/M
or increase aeration basin DO concentration. However,
while conceptually straightforward, the implementation of
these stepsin practice can be difficult, fraught with unde-
sirableconsequences, and uneconomical.For example:
Control of ActivatedSludgeBulkingand Other SettlingProblems
The ability to raise DO concentration may be limited
by the available aeration capacity.
. Raising DO could cause the onset of nitrifica-
tion or, worse still, partial nitrification. Both of
thesefurther increasethe oxygen requirements,
and partial nitrification causesrapid denitrifica-
tion in the secondary clarifier and increased
chlorine demand for effluent disinfection.
. Lowering F/M could causenitrification and the
problemsdiscussedabove.
. Lowering F/N4 will increase MCRT causing
increasedoxygen demand for endogenousres-
piration.
. Lowering MCRT requires an increaseof MLSS
concentration to a level that could exceed the
secondaryclarifier solids loading capacity. It is
difficult to do this when the clarifier blanket is
already so high that the clarifier is overflowing
solids.
These problems are especially critical for municipal
wastewatertreatmentplants that treat dilute wastewaterin
well mixed aeration basins at temperaturesabove about
23"C and operatein a nonnitrifying mode. Such plants are
unstable becausethey continually juggle MCRT and aer-
ation basin DO to prevent low DO filament growth and
nitrification. Because of these factors it is necessaryto
consideralternatives:
. Continue operating at low aeration basin DO
concentration and control the filamentous
organisms by chlorination.
. Consider modifying the well mixed aeration
basin to an anaerobicor anoxic selectorconfig-
uration. These types of selectorscan control all
types of low DO filamentousorganisms.
2. Case Histories
a. Orange County Sanitation District Plant,
Fountain Valley, CA
This high purity oxygen activated sludge plant treated
mixed municipal and industrial wastewaterscontaining,
on occasion, citrus-processingwastewater.DO uptake
ratesin the first aerationbasin compartmentreachedvalues
of approximately 2OOmg Oy'g VSS/h. When the plant was
operated at DO levels of l0 to 12 mglL, bulking due to
the low DO Type 1701 occurred even though the DO was
l0 to 12 mg/L because the very high DO uptake rates
required very high bulk DO concentrationsto prevent the
growth of low DO filaments. When the DO concentration
109
400
a
o
ta
d '
,l
E aa
i 200 ooo
o
a
a atat '
3O
0102030
DO uptakerate,mg O2lgVSS,h
FIGURE 4.33 Effect of dissolvedoxygen uptake rate on SVI
of a pulp and paper wastewateractivated sludge treatment plant.
Monthly averagedata.
was raised to 16 to 20 mg/l-, the levels of Type l70l
decreased and sludge settling improved.
b. Pulp and Paper Wastewater Activated
Sludge Plant
The SVI was relatedto its DO uptakerate (Figure 4.33).
The filamentousorganismscausingthe settling problems
were largely the low DO types: H. hydrossis and Type
1701.Figure4.33 showsthat SVI increasedapproximately
linearly with DO uptake rate. As observed earlier, SVI
increasedas the mixed liquor temperatureincreased.This
observationis consistentwith the occurrenceof low DO
bulking becausetemperature causesthe DO uptake rate
to increase and the increasedDO uptake rate causeslow
DO filamentous organisms to grow.
c. City of Woonsocket, Rl
An interesting approach combining chlorination for low
DO bulking control and energy savings for aeration was
usedby Banoub(1982) at this l6-MGD (0.69 m3/s)design
flow activated sludge plant. At the time of Banoub's work,
plant flow was 8.5 MGD (0.42 m3/s).Becauseof the high
cost ofelectrical energy($0.08/kwhincluding fuel adjust-
ment) it was decided to economize on aeration energy by
maintaining low aeration basin DO. This encouragedthe
growth of low DO filamentous organisms and increased
SVI. RAS chlorination at 3 kg CI2/103kg MLSS/d was
used to reduce the SVI to below the target value of
225 mL/g on the few occasions when it was exceeded.
1 10 M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Chlorination took "a couple of days" to reduce the SVI
to the target value.
Previous operation to maintain an aeration basin DO
of 2.0 mg/L required the operation of one aeration blower
at maximum amperage(330 amps) during the winter and
an additional aeration blower during the summer. When
operatingat low DO (0.2 to 0.5 mg/L), only one aeration
blower operating at its minimum amperage (240 amps)
was required year-round,resulting in an annual savings
of more than $41,000(averageof $1 12ld). On days when
RAS chlorination was necessary,approximately 85 lb
(39kg) Clrld was usedat a daily cost of lessthan $10.
Banoub(1982) pointed out that extremecare must be
taken when operatingin sucha mode.Frequentsettleabil-
ity monitoring and microscopicexaminationsof the acti-
vated sludge were made. Banoub also pointed out the
exceptionallyhigh effluentquality (on the order of 5 mg/L
each of BOD. and SS and turbidity of 2 FTU) attainable
from a high SVI sludge.It should be remembered,how-
ever,that this plant was receiving approximately50Voof
its designflow so that amplesecondaryclarificationcapac-
ity was availableto accommodatethe high SVI sludge.
C. Errrcr oF AERATToN
Blsrru CorrrcuRATroN,
Wasrrwarrn FrrorrucMrrHoo, AND REDox
CoNorrrons oN AcnvATEDSruDcE SrrrlrNc
Cnnmcrrnrsrcs
1. Effect of Aeration Basin Configuration
In general,activatedsludgesystemsthat are continuously
fed and have well mixed aerationbasinsthat are always
aeratedthroughoutwill producepoorer settling activated
sludge than systemsthat are fed intermittently or have
compartmentalizedaeration basins and relatively high
wastewaterconcentrationat the point where RAS and
influent wastewatermix (Heide and Pasveer,1974; Ren-
sink et al., 1982;Tomlinson,1982;and Chiesaand Irvine,
1985). Furthermore,activatedsludge settling usually is
poorer if this initial mixing or contactzone betweenRAS
and influent wastewatercontains DO and/or is aerated
rather than being devoid of DO and/or unaerated(Chiesa
and Irvine. 1985).
Tomlinsonand Chambers(1978b)conducteda survey
of full-scale activatedsludge plants in Great Britain and
measuredSSVI35 and the distribution of hydraulic resi-
dencetimes in the aerationbasins(not including second-
ary clarifiers and RAS streams).The tracer study results
for hydraulic residencetime distribution were interpreted
using a "completely mixed tanks in series" model (one
tank in series representeda completely mixed basin and
an infinite number of tanks in seriesrepresenteda plug
flow basin). As the degree of plug flow increased,the
SSVI3s generallytendedtoward lower, less erratic values
(Figure 4.34). Heide and Pasveer(1914), working with
Pr oblem s
300
g 2oo
-.,1
a
n o
2 too 3t.t N=rol--:>
'a'
o
0r0203040s0
Number
of equivalent
tanks,N
FIGURE4.34 RelationshipbetweenSSVI3.'andaeration
basin
(FromTomlinson,
mixingcharacteristics. E.J.andChambers, B.
(1978b),TechnicalReportTRl22, WaterResearch Centre,
U.K. With permission.)
Stevenage,
low loaded, completely nitrifying oxidation ditches and a
24-h fill- and-draw laboratory batch activated sludge unit,
showedthat the SVI was 500 to 600 mllg for a continuously
fed oxidation ditch, 100 to 150 mllg for an oxidation ditch
fed discontinuously,and 40 to 50 ml-/g for the laboratory
batch unit fed once per day. When an oxidation ditch was
precededby a small anoxic mixing tank (averagehydraulic
detentiontime = 15 min) through which both the RAS and
wastewaterpassed,the SVI was less than 70 ml/g.
The occurrenceof bulking in completely mixed sys-
tems and not in plug-flow or intermittently fed systems
has been explained in various ways. Chudoba et al.
(1913a) proposed that filamentous organisms possess
lower half-saturationcoefficients(K.) than floc-forming
microorganismsfor organiccarbonsubstrates. This allows
them to be selectedin completelymixed, low F/lM systems
where the carbonaceoussubstrateconcentrationis low
throughout the aerationbasin since the influent wastewater
is dispersedthroughoutits entire volume.This hypothesis
cannot completely account for the suppressionof bulking
achievedby allowing activatedsludge to pass through a
short hydraulic residencetime zone ofhigh carbonaceous
substrateconcentrationbefore entering the aerationbasin.
It hasbeenproposedthat the effectiveness of suchsystems
in suppressing bulking is that they favor the growth of
floc-forming organisms that have higher substrateuptake
rates and storage capacities (a transient, unbalanced
growth response) than filamentous organisms (van den
Eynde et al., 1982b; Grau et a1.,1982) or floc formers that
have greater starvation resistancethan filamentous organ-
isms (Chiesaand Irvine, 1985).
Control of bulking has been obtained consistently in
laboratory studies by compartmentalizing aeration basins
to approach plug flow (Chudoba et al., 1973a;1913b;
1974; Rensink et a1., 1982; Spector, 1977), intermittent
feeding of wastewaters(Houtmeyers,1978; Houtmeyers
 Control of ActivatedSludgeBulkingand Other SettlingProblems
Influent
wastewater
sludgeconfiguration.
FIGURE4.35 Selector-activated
et al., 1980;Verachtertet al., 1980; van den Eynde et al.,
1982a; 1982b),use of small mixing tanks where RAS and
influent wastewater mix prior to reaching the aeration
basin (Lee et al., 1982; Grau et al., 1982), or fed-batch
operation(Goronszy,1979; Goronszy and Barnes, 1979;
Y
l-
Barnesand Goronszy,1980;Chiesaand Irvine, 1985).All
thesemeasuresproducea carbonaceous substrateconcen-
tration gradient in the aeration basin or a high substrate
concentration at the point where RAS and influent waste-
water enterthe aerationbasinsystem,followed by a close-
to-zero soluble substrate concentration thereafter in the
aerationbasin.
2. Selectors
Chudoba et al. (1973b) devised the term selector to
describean activatedsludgesystemcontainingsmall sep-
arateinitial mixing zonesfor RAS and influent wastewater
(Figure 4.35).The term refersto the role of such a device
in selectingactivatedsludgeorganismswith desirableset-
tling characteristics.The need to compartmentalizethese
initial contact zones was stressed.The beneficial effects
of anaerobic or anoxic conditions in the initial contact
zones were recognized in some of the early work on the
effect of aeration basin configuration on activated sludge
settling.For the purposesof this discussion,the following
definitions will be used'
. Aerobic or Oxic - DO is presentand supplied
in sufficient quantities to meet metabolic
demands. Storage and aerobic respiration are
the primary mechanisms of soluble substrate
removal.
o [n61lg - DO is absent;NO.-N is presentand
supplied in sufficient quantities to meet meta-
bolic demands. Storage and denitrification are
the primary mechanisms of soluble substrate
removal.
. Anaerobic - DO and NO,-N are both absent.
Polyhydroxyalkanoate (PHA) storage coupled
111
Secondary
clarifier
with hydrolysis of stored inorganic polyphos-
phate or fermentation of stored glycogen are
the major metabolicactivitiesremoving soluble
substrate.
The aeration basin configuration of initial anaerobic
zonesfollowed by aeratedor oxic zonesis also used for
enhancedbiological phosphorusremoval (EBPR) by acti-
vated sludge.Indeed one EBPR patenttitled "Production
of Non-Bulking Activated Sludge" (Spector, 1977) pro-
videsthe basisfor the A/O process- a configurationthat
usually producesactivatedsludge with excellent settling
properties(Honget al., 1984;Deakyneet al., 1983).Tracy
et al. (1986) showedthat initial anaerobicconditionspro-
vided a further improvementof settlingpropertiesbeyond
that afforded by aeration basin compartmentalization.
They operatedfill-and-draw activatedsludge systemsin
which the feedingconditionswere anaerobic,aerobic,and
oxygen-limited.Feedperiodswere all followed by aerobic
periods, settling, and effluent withdrawal. With initial
anaerobic contact, the SVI averaged55 to 65 ml-/g; with
the initial aerobic contact period, the SVI averaged
100 ml/g; when the initial contact period was oxygen-
limited, the SVI averaged300 ml/g, (most likely due to
low DO filamentous bulking). Similar results were
obtained by Watanabeet al. (1984); Chiesa and Irvine,
(1985)and Inamori et al. (1986).
Even if the conditions for EBPR do not exist in an
activatedsludgesystem,solublesubstratecan be removed
and storedin an initial anaerobiczoneby G bacteria(Cech
and Hartman, 1993:Liu et al., 1996 1997:Nielsen et al.,
1999; Seviour et al., 2000). These organismsuse energy
produced by the fermentation of stored glycogen to drive
the anaerobic uptake and storage of soluble substrate
(Mino et al., 1998).
Initial anoxic zones were found to control bulking
when NO,-N was generated in the aerated part of the
aerationbasin(Cooperet al., 1977).Tomlinsonand Cham-
bers (1979) concludedthat an initial anoxic zone placed
in a nitrifying activated sludge system improved settling
112 M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
properties by being anoxic and by reducing longitudinal
mixing. Wagner (1982) improved activatedsludge settling
in nitrifying activated sludge by introducing a predenitri-
fication reactor. Hoffman (1987) showed that compart-
mentalized aeration basins with initial anoxic zones pro-
duced better settling activated sludge than an equally
compartmentalized,totally oxic system.Chiesa and lrvine
(1985) and Ip et al. (1987) demonstratedfilamentous
organism suppressionin sequencing batch reactors with
anoxic fill periods.
3. Selector Effect
a. GeneralObservations
When activatedsludgeis subjectedto a sequenceof envi-
ronmental conditions where it is first fed and then starved
as in a compartmentalized reactor, a batch fill-and-draw
system,or a selectorsystem,the microbial make-upof the
sludgeadjuststo the situationby developingthe ability to
rapidly take up soluble substrateand store it internally for
use during the starvationperiod. This soluble substrate
uptakeand storageability is the basisofthe selectoreffect
becausemany filamentous organismsare less able to per-
form in this way than some members of the floc-forming
community.The selectoreffect is illustratedin Figure4.36
and Figure 4.37.
The curvescomparethe time courseof solublesubstrate
concentration(soluble COD) and respiration rate (DO
uptake rate) in a batch test on activatedsludge taken from
a selector activated sludge system and from a completely
mixed (CSTR) activatedsludgesystem(van Niekerk et al.,
1987).The selectoractivatedsludgetakesup solubleCOD
250
J
d )nn
.-i
d
o l {n
x
O
E r no
E
4
.E
.E
tr <fl
Time.h
FIGURE 4.36 Soluble chemical oxygen demand uptake during batch substrate uptake experiments with completely mixed and
aerobic selector activated sludges. (From van Niekerk, A.M. et al. (1987), J. Water Pollut. Control Fed., 59,262. Wirh permission.)
Pr oblem s
much faster than the CSTR activatedsludge (Figure 4.36).
The DO uptake rate increaseswhile the rapid soluble COD
uptake is occurring in the selector activated sludge
(Figure4.37). Measurementsof the total mass of soluble
COD taken up and the total mass of DO consumedby the
selector activated sludge during this initial period show
that only a small fraction of the soluble COD that disap-
pears from solution (usually less than 257o and often as
little as l07o) can be accountedfor by oxygen consump-
tion. This meansthat most of this soluble COD is stored
rather than oxidized.
b. Selector Mechanisms
It is important to understandthat selectorsystemsnot only
select againsl some types of filamentous organism, but
also that they select/or certain types of organisms(mostly
nonfilamentous) that can take up soluble substraterapidly
and store it. To survive in a selector system (and therefore
be selected)an organism must have a high soluble sub-
strate uptake rate and large substrate storage capacity.
These characteristicsare not evident in the floc-forming
microorganismsthat grow in a completely-mixed, contin-
uously fed activatedsludge system.
Therefore,it must be assumedthat selectorschange
the metabolisms(and possibly the types) of floc-forming
organismsand selectagainstfilamentousorganisms.Evi-
denceior this was providedby van Niekerk et al. (1987b)
who showedthat, in aerobic selectors,large numbers of
amorphouszoogloealcoloniesdeveloped.Thesecolonies
were not seenin CSTR-activatedsludge;the colonieswere
isolatedand shown to have high soluble substrateuptake
rates and large substratestoragecapacities.
o Completely mixed
Aerobic selectors
1.5 3.0
Control of ActivatedSludgeBulkingand Other SettlingProblems 113

O-O Completely mixed

60 A--{ Aerobic selectors

ui
U)

*4 5
oo

JI JU
;

0 0.5 1.0 1.5 2.0 2.5 3.0

Time,h

FIGURE 4.37 Respiration rates during batch substrate uptake experiments with completely mixed and aerobic selector activated
sludges.(From van Niekerk, A.M. et al. (1987), J. Water Pollut. Control Fed., 59,262.Wirh permission.)

selector-activatedsludge. Type 021N is a filamentous

organism that grows well on acetateand can causebulking
in activated sludge treating wastewaterscontaining high
soluble COD concentrations(such as septic sewage,food
l6 processing wastewaters and brewery wastewater). The
;
acetateuptake rate of Z. ramigera is higher than that of
U)
U)
Type 02lN over most of the growth rate range tested.
Figure 4.38 predicts that under intermittent feeding con-
I ro
o ditions (typical of a selector), Z. ramigera almost always
oo would be able to take up and sequesterthe acetatebefore
'Ilpe 02lN
Type 021N had a chanceto get at it. The picture is quite
e
E different when acetate is fed continuously (typical of a
6a
steady-stateCSTR-activated sludge system).
q
Figure 4.39 shows that at low acetateconcentrations,
Type 021N can out-compete Z. ramigera. van Niekerk
44 et al. (1987) showed that these competitive advantages
could determine which of these organisms dominated in
a mixed chemostatculture grown at a steady state growth
rate designed to favor the Type 021N filamentous organ-
ism. When low frequency, intermittent feeding of acetate
0 0.2 0.4 0.6 0.8 1.0 was employed, Z. ramigera dominated (nonbulking).
growthrate,p"/Irnax
Dimensionless When the feeding was continuous or intermittent with a
short interval between feedings, Type 021N dominated
F|GURE4.38 Maximum acetateuptakeratesof Z. ramigera (bulking) (Figure 4.40).
and Type 02lN in batchtests.(From van Niekerk,A.M. et al.
(1987),J. WaterPollut.ControlFed.,59,262.
Withpermission.) The precedingdiscussionof selectormechanismssug-
gests that selectorsonly influence the uptake and storage
Figure 4.38 illustrates the abilities of two types of of soluble substrate.With one currently known exception
organismsisolated from activatedsludgeto take up acetate (see Chapter 5), it appears that selectors function by
(a simple, soluble substrate)in batch culture - conditions removing low molecular weight, readily metabolizable
that simulate a selectorreactor (van Niekerk et al., 1987). organic substancesthat can be readily transported across
Z. ramigera is a floc-forming bacterium found in aerobic cell walls. This point is illustrated in Figure 4.41 and
114 M anua lo n C a u s e sa n d C o n tro lo f A c ti v atedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
P r oblem s

soluble substrateis measuredas soluble COD (0.45 pm

filtered COD), a selector effluent concentration of up to
about 60 mg/L will allow an SVI of <100 ml-/g to be
achieved (Figure 4.41). However, if the low molecular
weight readily transportable and metabolizable organic
componentsof the soluble COD are measuredin the selec-
tor effluent,one seesthat thesemust be almostcompletely
*4 Type021N
E removed(<1 mg/L) to achievean SVI of 100mllg (Figure
4 4)\
When organic substratesare transported into micro-
E3
bial cells and transformed into storage products, sources
d
of energy and reducingpower are required.Selectorscan
o be designatedaerobic,anoxic, or anaerobicon the basis
a-
of the method of achievingthis oxidation. If DO is used,
the selector is aerobic and aerobic respiration occurs
(Figure4.43); if NO..-N is used,the selectoris anoxic and
denitrificationoccurs(Figure 4.44). lf hydrolysisof inter-
nally storedhigh energyinorganicpolyphosphateis used,
the selector is anaerobic and release of orthophosphate
occurs (Figure 4.45); and if storedglycogenis fermented
0 12 34 5
to produceenergy,the selectoris anaerobic(Figure 4.46).
substrateconcentration,
mg acetate/L
When inorganic polyphosphatehydrolysis provides
FIGURE 4.39 Predictedrelationshipof steadystategrowth rate the energyfor substrateuptakeand storage,EBPR occurs
and acetateconcentrationfor Type 02 1N and Z. ramigera. (From becausethe anaerobicallyreleasedorthophosphateand
van Niekerk, A.M. et al. (1988), J. Water Pollut. Control Fed., influent orthophosphateare taken up aerobically in the
60, 100. With permission.) main aerationbasin using energygeneratedfrom the oxi-
dation of the stored substrate.When glycogen provides
the energy for anaerobicsubstrateuptake and storage,the
Feedingfrequency,times ld
A processcompeteswith the polyphosphate-based reactions
describedabove and consequentlyinterfereswith EBPR.
In effect, it robs the EBPR organismsof substrateso less
1 00
is available for performing the reactions necessaryfor
s EBPR.
The selectorsubstrateuptake mechanismsdiscussed
E?5 above are virtually mutually exclusive because of the
highly competitive environment that exists in activated
E
sludge.The energyreleasedby oxidizing an organic sub-
strateby aerobic oxidation (where Or is the terminal elec-
5)U
tron acceptor)is greaterthan for denitrification (where NO.
E
is the terminal electron acceptor),and in tum this is greater
o----4 Z. ramigera
than for the polyphosphatehydrolysis and glycogen fer-
25 mentation carried out respectivelyby the EBPR organisms
o_r Type02lN
and G bacteria under anaerobicselector conditions. Thus,
I
I
I
in the presenceof DO, a mixed microbial community such
as activated sludge will become dominated by microor-
0 48 12 16 20 ganisms that perform aerobic oxidations. Denitrification
Numberof MCRTs will not occur, even if NOr is present,becauseit is less
efficient than aerobic oxidation. Denitrification will only
FIGURE 4.40 Dual culture chemostatgrowth of Z. ramigera
occur when no DO is available.Likewise, the presenceof
and Type 02lN with intermittent feeding of carbon substrate.
(From van Niekerk, A.M. et al. (1988), J. WaterPollut. Control both DO and NO, will suppresspolyphosphate storage,
Fed.,60, 100.With permission.) hydrolysis,and glycogen fermentationmetabolism.
In many instances, however, more than one type of
Figure 4.42 (Shao and Jenkins, 1989) that show the rela- metabolism can occur in different portions of the same
tionship between the soluble subsffate concentration leaving basin. For example,both aerobic and anoxic oxidations
an anoxic selector and the SVI of the activated sludee. When can occur in an initial mixins cell that is not uniformly
Control of ActivatedSludgeBulkingand Other SettlingProblems 115

Irgend
a ShaoandJenkins(1988),20'C aa tl
500 I Shao and Jenkins(1988),25'C
A Daigger et al. (1985)
A Wheeler et al. (1983)
400 O Lee et al. (1982)
d
u lomllnson ano LnmDers I tyly,

i 300 On
'l
C)
a
200
a
ta
100 a
.

0 25 50 75 100 t25 150 175

SolubleCOD, mg/L

FIGURE 4.41 SVI as a function of anoxic selector effluent soluble COD concentration. (From Shao, Y.J. and Jenkins, D. (1989),
Water Sci. Technol.,2l:ll, 609. With permission.)

700
trgend
I Acetare(ShaoildJenkins. 19891
O Acetate (Chambers, 1982)
500 A Glucose (Shao and Jenkins, 1989)

) 400
Fi
>
6
300 A

200 ,r l
E
100

s0 100 150 200

Acetateor glucoseconcentration,
mg/L

F|GURE4.42 SVlasafunctionofanoxicselectoreffluentacetateorglucoseconcentration.(FromShao,Y.J.andJenkins,D.(1989),
Water Sci. Technol.,2l:11, 609. With permission.)

aerated. Likewise, anoxic metabolism may occur in the
upstream portion of a mixed unaerated initial contact
basin, and anaerobic metabolism may occur in the down-
stream portion if the supplied NO3-N is exhausted and
readily degradable soluble substrate remains. The acti-
vated sludge floc itself may provide an environment where
more than one type of metabolism can occur concurrently.
For example, in a highly loaded, poorly aerated initial
contact zone, aerobic oxidation can occur in the bulk
liquid and at the floc surface while denitrification, and
possibly anaerobicphosphaterelease,take place inside the
activated sludge floc.
Another aspect of the type of energy metabolism
occuring in the selector is its effect on the selection pro-
cess itself. Most activated sludge filamentous organisms
are aerobes.Thus, in an aerobic selector,selection occurs
only becauseof the relative abilities of the microorganisms
to rapidly take up and store substrate.This type of selec-
tion has been called kinetic. When anoxic or anaerobic
conditions are imposed on the selector,the kinetic selec-
tion is supplementedby a "metabolic" selection, i.e., the
ability to denitrify, store, and hydrolyze intracellular inor-
ganic polyphosphate or the ability to store and ferment
intracellular glycogen. The denitrification rates of some
filamentous organismsare much lower than those of floc-
forming bacteria (2. ramigera) growing in selectors(Thble
4.8; Shaoand Jenkins,1989;Blackall et al., 1991).More-
over, someof thesefilamentousorganisms(e.g.,Type 021N
and G. amarae) only reduce nitrate to nitrite rather than to
nitrogen gas, as Z. ramigera does (Blackall et al., 1991).
Some types of floc-forming bacteria (e.g., Rhodocy-
clus-like organisms; Zllles et al., 2002) can take up and
store substrateusing inorganic polyphosphatehydrolysis
for energy generation. It is suspectedthat many types of
116
M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Aerobic selector
SolubleCOD
SolubleCOD
CO2+ HrO
FTGURE4.43 Aerobic selector activated sludge substrateuptake and utilization mechanisms.
Anoxic selector
SolubleCOD
SolubleCOD
NO:
N,
CO2+ HrO
FIGURE 4.44 Anoxic selectoractivatedsludge substrateuptake and utilization mechanisms,
filamentousorganismsare unable to do this. Blackall et
al. (1991) showedthat G. amarae could not take up sub-
strateanaerobically.
In discussingthe selectoreffect, we have made refer-
ence only to eventsocculring in the initial contact zone.
For proper functioning, events that occur in the aeration
basin also are very important.The fact that carbonstorage
takes place in the selector can be demonstratedin several
ways. Shao and Jenkins(1989) measuredthe decayrates
of various types of activated sludge activity for sludges
developedin anoxic selectorsand CSTR systemswhen
they were aeratedin the absenceof exogenous substrate
(as they would be in the aerationbasin following a selec-
tor). As Table4.5 shows,the anoxic selectorsludgeactiv-
ities persistedlongerthanin the CSTR sludges.The MLSS
decay rate for anoxic selector sludge also was lower than
for CSTR sludge.Theseobservationssuggestthat storage
products allow the selector sludge floc formers to survive
for longer periods in the absenceof exogenoussubstrate
than the CSTR floc formers.
In addition to the ability to better survive in the aer-
ation basin under starvationconditions, the organismsthat
take up and store substratein the selector must be able to
utilize their stored products in the main aeration basin. If
this does not occur, the organisms will be returned to the
Pr oblem s
Main aeration basin
CO, + H2O
Cell wall
Main aeration basin
CO2+ HrO
Cell wall
selectorwith diminishedstoragecapacity.Eventuallythis
situationcould lead to saturationof storagecapacity and
soluble organic substratepassingthrough the selectorto
the aeration basin. If this occurred, the feed-and-starve
cycle requiredfor the selectoreffect would be eliminated
and filamentousorganismgrowth could be expected.It is
therefore important to size a selector correctly to encour-
age rapid uptakeof the dissolvedorganiccompoundsand
also designthe aerationbasin so that it can provide proper
conditionsfor utilizing the substratestoredby the micro-
organisms.Becausean anoxic selectorrequiresnitrate,the
mixed liquor (containingnitrate) must be recycledto the
upstreamanoxic selector.Therefore, the controlling crite-
rion for sizing the main aeration basin is to provide an
aerobic MCRT (solids inventory in the aerobic zone
divided by process solids wasting rate) that will allow
nitrifying microorganisms to become established (Grady
et al., 1999).
The successfulfunctioning of an anaerobicselector in
which inorganic polyphosphate hydrolysis (EBPR) takes
place requires main aeration basin conditions that favor
the growth of microorganisms capable of polyphosphate
storage. These microorganisms grow more slowly than
other heterotrophic microorganisms(Mamais and Jenkins,
1992).
Cont r olof A c t i v a te dS l u d g eBu l k i n ga n d Oth er S ettl i ngP robl ems 117

Anaerobic selector Main aeration basin

COr + HrO

Po+

Cellwall
Cell wall

FIGURE 4.45 Anaerobic selectoractivatedsludgesubstrateuptake and utilization mechanisms involving polyphosphate hydrolysis
and synthesis.

Anaerobic selector Main aeration basin

SolubleCOD CO2+H20

Cell synthesis

Cell wall

tIGURE 4.45 Anaerobicselectoractivatedsludgesubstrateuptakeand utilization mechanismsinvolving glycogenfermentationand

synthesis.

TABTE4.5
Comparisonof Decay Ratesfor ActivatedSludgefrom CompletelyMixed
and Anoxic SelectorSystemsunder StarvationConditions
FirstOrder Decay Rate(d-t)
ActivatedSludgeType MLSS COD Uptake Rate Oxygen Uptake Rate NO3-N Uptake Rate

CSTRsystem 0.050 0.39 0.38 0.57

Anoxicselector
system 0.034 0.25 0.22 0.29

Source: From Shao,Y.J. and Jenkins,D. (1989), Water Sci. Technol.,21:'11,609.With permission.

The growth rates of both the nitrifiers and EBPR
organisms are influenced by temperature.Figure 5.23
shows the effect of temperature on the minimum (wash-
out) aerobicMCRT for both groups.Actual aerobic MCRT
valuesused for selectorsystemdesign will be somewhat
longer than these minimum values to allow development
of an adequate and stable population of the required
microorganisms.
It is interesting to note that as the temperature
increases,the zone of the MCRT in which one can obtain
EBPR without nitrification diminishes markedly. For tem-
peraturesof 26"C or higher, it is almost impossible to have
EBPR without at least some nitrification. The implication
of this for selectors is that at higher temperatures the
selectormechanismmay well be a mixture of anaerobic
and anoxic processes.
c. Effects of Selectors
From the previousdiscussion,it can be deducedthat selec-
tors should be generally effective against filamentous
organismsthat grow on simplesolubleorganiccompounds
(such as are removedin selectors),filamentousorganisms
that do not have high soluble substrateuptake rates, and
filamentous organisms that do not have high substrate
storagecapacities.In addition, filamentous organismsthat
do not denitrify rapidly or completely (or at all) will be
118
M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
TABLE4.6
of Selectorsin Controlling
Effectiveness
FilamentousOrganisms
Effective Not Always Effective
S- natans Type0041
Type 1701 Type0675
Type02lN Type0092
Thiothrix spp.. M. panicelLa
N. limicola
H. hydrossis
Type 1851
Nocardioform organisms--
- Not when caused by nutrient deficiency.
-'Aerobic selectorsnot alwayseffective;anoxicselec-
tors effective.
Source:From Cha, D.K. et al. (1990), WaterEnviron.
R e s . ,6 4 , 3 7 .
out-competedin anoxic selectorsand those that are unable
to take up substratecoupled with anaerobic inorganic
polyphosphatehydrolysis or glycogen fermentationwill
be out-competedin anaerobicselectors.No type of selec-
tor is effective againstthe growth of filamentousorgan-
isms causedby low pH, septicor other sulfide-containing
wastewaters,and nutrient (N and P) deficiencies.
Table 4.6 lists the types of filamentous organisms
against which selectorshave been successful.The table
also lists filamentousorganismsagainstwhich selectors
are not consistentlyeffective.The reasonsfor the inability
of selectorsto control these filamentousorganismswill
be discussedat the end of this section.
4. Selector Design (Sizing)
a. Ceneral
Early approachesto selectordesigninvolved using selec-
tors sized on the basis of a certain fractional volume of
the aeration basin. This was derived from the work of
investigatorssuch as Lee et al. (1982) who showed that
activated sludge settling characteristicswere functions of
fractional selectorsize (Figure 4.47). This approachhas
been refined considerably in recent years. The general
criteria presentedbelow may be used when site-specific
data are not available. However, specific experience and
data will allow refinement of the selector design and the
performance achieved.
b. lnitial Contact Zones
The initial contact zone organic loading or (FAvI)' should
be high enough to produce rapid soluble organic matter
uptake rates.Especially useful in this regardis the work of
Tomlinson (1916) showing the relationship between SVI
Pr oblem s
------o---\
{
Max SVI 2.5
^" 300
J
;
s 200
Ch
100
0 20 40 60 80 100
FIGURE4.47 Relationship of SVI'., to relativesizeof initial
aerationbasincompartment. V, = total aerationbasinvolume;
V, = initial aerationbasinvolume.(FromLee,S.-E.et aL.(1982),
WaterSci.Technol., 146-7,407.With permission.)
and initial contactzoneorganicloading (Figure 4.48).These
data suggest that (F/l\4)' values greater than about 3 kg
BODr/kg MLVSS/d will produceSVIs of 100to 150mllg.
The selectorshould be sized to allow sufficienttime
to remove all soluble,readily metabolizableorganicmat-
ter. Since it may not be possibleto measurethe amount
of this type of organic matter, soluble COD is used as a
surogate parameter.The data of Shaoand Jenkins(1989)
suggestthat (for a municipal wastewater)a selector efflu-
ent soluble COD of approximately 6O mglL will produce
an SVI of about 100 ml-/g (Figure 4.41). Selectordesign
practice in Czechoslovakiaand Austria (Chudoba and
Wanner, 1987) usesa COD removal criterion of 80% of
the removable COD (defined as influent soluble COD
minus secondaryeffluent soluble COD).
The selectorshould consistof at least three compart-
ments. This establishesa substrategradient (enhances
kinetic selection), reduces longitudinal mixing, and
accommodateswastewaterflow and strength variations.
c. Aerobic Selectors
For aerobic selectors the initial contact zones should be
sized so that first compartmentF/lM = 10 to l2kg COD/kg
MLSS/d and overall selector FAyI = 3 to 4 kg COD/kg
MLSS/d. An-effective alrangementthat yields these load-
ing relationships is a three-compartmentselector with the
first two compartments equal in size and the third com-
partment double the size of the first two. Initial contact
zone organic loadings much in excess of these values
should be avoided (especially with wastewatercontaining
significant amounts of readily available organic matter)
because viscous activated sludge may be produced. For
example, in the first compartmentsof aerobic selectorsin
plants treating brewery wastewaterand soft-drink bottling
wastewater,DO uptake rates in excess of 200 mg Or/g
VSS/h were produced and viscous (slime) bulking
Cont r olof A c t iv a te dS l u d g eBu l k i n ga n d O th e r S ettl i ngP robl ems
700
a
600
a
a
500 a
{ aoo a
.:.
-a
Fi
t
ao
>
(D 300 t r
-o
-a
200 aa
aa
faa
100
a' aa- '
FICURE 4.48 Effect of initial contact zone F/M on SVI for aeratedinitial contactzones.(From Tomlinson,E.J. (1976), Technical
Report TR35, Water ResearchCentre, Stevenage,U.K. With permission.)
resulted.Bypassing some of the wastewateraround the
selectoreliminatedthe viscousbulking.
For aerobicselectors,a supply of oxygen is required.
While DO uptakeratesof 50 to 60 mg O2/gMLVSS/h or
more may be encountered,the amountof oxygenrequired
is only a small fraction of the soluble COD removed-
typically 15 to 25Vo.The DO concentrationshould be
b et weenl and2 mg l L .
d. Anoxic Selectors
The (FAvI), values for anoxic selectors can be reduced
becausethe selectorutilizes both kinetic and metabolic
selection (ability to denitrify) to combat filamentous
organisms.To optimize denitrification and selection, a
selectorshould have initial contact zone loadings as fol-
lows: first compartmentFAvI = 6 kg COD/kg MLSS/d and
overall selectorFAvI= 1.5 kg COD/kg MLSS/d. An effec-
tive arrangementthat providestheseloadingrelationships
is a three-compartment selector with the first two com-
partments equal in size and the third compartment double
the size of the first two.
If only a singlebasin is availablefor use as an anoxic
selector, its FA4 should be <1 kg BODr/kg MLSS/d for
wastewatertemperaturesof <18"C and <1.5 kg BODr/kg
MLSS/d for wastewater temperaturesof >l8oC (Marten
and Daigger, 1997).For singlecompartmentanoxic selec-
tors, an alternative method of sizing is to use anoxic
MCRT. To calculate anoxic MCRT, divide the mass of
MLSS in the anoxic selector by the total mass of SS
wasted each day. For single basin anoxic selectors,the
anoxic MCRT should be in the range of I to 2 d (Marten
and Daigger, 1997: Clark et al., 2001).
Becausean anoxic selectorremovessoluble COD by
denitrification, its DO concentration must be low. The
selector contents should be mixed by mixers, very low
119
a
t a
o
a
a
a o I I- ar lo a
olo o o
a
ota
a
Initialcontactzone(F/M)i,kg BOD5/kgVSS,d
aerationrates,and/orinefficientaerationdevices.The acti-
vated sludge system must nitrify and sufficient NO3-N
must be introducedinto the selectorin the RAS streamor
by internalmixed liquor recycle.If sufficientNO.-N can-
not be introducedinto the selectorto removethe necessary
soluble COD, then the selectoreffect can be achievedby
making only part of the selector anoxic. Typically, the
initial portion of the selectorwould be anoxic and utilize
the supplied NO.,-N; the downstreamportion would be
anaerobicdue to the depletion of the supplied NO.,-N.
Alternatively,if the first part of the selectoris anoxic, the
secondpart may be aerobic.In anotheroption, if the first
part of the selectoris anaerobic(i.e., containsonly RAS
which is very low in NO.-N and influent wastewater),the
secondpart can be anoxic with mixed liquor recyclecould
be introducedinto the anoxiccompartment.
Due to variationsin the relative proportionsof COD
that are oxidized and stored, values for the amount of
soluble COD that can be removedby I mg NO.-N vary
considerably.For influent domestic wastewatera typical
value is 8 mg soluble COD/mg NO3-N if little or no
storageoccurs (essentiallyall soluble COD is being
metabolized).Higher valueswill be observedif significant
storageoccurs.
The overall sizing of an anoxic selectormust allow
sufficient time for denitrification to take place to such a
degreethat it reducesthe soluble COD to the targetselector
effluentvalue (e.g.,60 mg solubleCOD/L). Denitrification
rates for the readily metabolizable soluble component of
influent domestic wastewater- the material required to
be removed in a selector typically on the order of
5 to 10 mg NO,-N/g MLSS/h at 20"C. Note that the
objective in an anoxic selectoris not the complete removal
of NO.,-N; it is the use of NO,-N to removesolubleCOD
down to a level of approximately60 mg/L. If denitrification
12O M anua lo n C a u s e sa n d C o n tro lo f Ac t i vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Pr oblem s

is an additional objective to bulking control, a greater

fraction of the initial zone of the aeration basin may have TABLE4.7
to be anoxic. As indicated, staging of the selector will Design,Operating Parameters, and Resultsof
enhancethe overall nitrogen removal rate and efficiency. SelectorActivatedSludgeProcessat the
Leopoldsdorf SugarMill, 1985 Season
e. Anaerobic Selectors
Anaerobic selectors utilize both kinetic and metabolic Parameter Unil Value

selectionprocesses.The overall sizing is dictatedby the Wastewater flow MGD 12

rate at which soluble organic matter is taken up and ortho- Selector volume MG 0 .1
phosphateis releasedunder anaerobic conditions. For Aeration basin volume MG 4.3
domestic wastewaters,the total anaerobicselectorhydrau- Selector hydraulic residence h 0.2
lic retentiontime is usually in the range of 0.75 to 2.0 h. tlme
The selectorshould be divided into at least three compart- Aeration basin hydraulic h 8.7
residence time
ments with the same(F/M), relationships as for the anoxic
MLS S rnglL 10,000
selector. For a completely anaerobic selector, DO and
MCRT d8
NO3-N must be excluded. For a nitrifying activated mL/g 50
SVI
sludge,this may require an anoxic zoneat some location COD removal in selector 7o of COD,,,,removed 72
in the flow scheme. Oxygen consumedin selector 7o of total oxygen 8.5
consumed
f. Main Aeration Basin Influent total COD mg/L 450
Conditions in the main aeration basins (aerobic zones) Effluent soluble COD mg/L 45
downstreamfrom the selectorcan also affect sludge set- Effluent total BOD mglL 340

tleability. SVI values will be minimized if the aerobic Effluent soluble BOD mglL 10

zones are compartmentalizedand if fully aerobic condi- Source: From Kroiss, H. (1985), Wien. Mitt. Band.,56, 81. With
tions (>1 to2mg DO/L) are maintained. permission.

5. Selector Case Histories

a, Leopoldsdorf Sugar Mill, Austria
This is an aerobicselectorcasehistory.In Czechoslovakia
and Austria, the following designcriteria apply to aerobic
selectordesign (Chudobaand Wanner, 1989):
Number of compartments:four, equal-sized
Total selectorF/M: 3 kg BODr/kg MLSS/d
Initial selectorcompartmentF/\4 or (F/M)': 12 kg
BODr/kg MLSS/d
Selectoroxygenationcapacity:4 kg Orlm3ld
Total systemF/M: 0.3 kg BOD./kg MLSS/d
MLSS: 3300 mg/L
Total systemoxygenationcapacity:2 kg Ot/m3/d
Using thesedesigncriteria,Kroiss (1985) designedan
aerobic selector for the Leopoldsdorf beet sugar mill in
Marchfield, Austria and obtainedthe resultsshown in Table
4.7 during the 1984 season.This activatedsludge system
had beenpreviously plaguedwith severebulking problems
due to Type 021N. The selector system had an (F/I\4)' of
approximately 4.0 kg BODrlkg MLSS/d (overall F/Ir4 of
0.10 to 0.15 kg BODr/kg MLSS/d). The selectorremoved
727o of the total COD, only 8.57o of which could be
accountedfor by oxygen uptake. SVI values of 50 ml-/g
were obtained at MLSS concentrationsof 10.000 ms/L.
b. Hamilton, OH
This casehistory describesaerobicand anoxic selectors.
The activatedsludgeplant is a 25-MGD (l.lm3/s) facility
that, at the time of this work, was treating approximately
19 MGD (0.8 m3/s)of wastewaterconsistingof approxi-
mately 7.5 MGD (0.3 m3/s)of domesticwastewater;the
remainder is paper manufacturing wastewater.
Following primary clarification, the primary effluent
is split between(l) a plant constructedin 1958consisting
of an aerated channel with a hydraulic retention time of
approximately 8 to 12 min in which a mixed RAS and
primary effluent stream flows prior to entering three dif-
fused air aerationbasins(T3) and (2) a plant constructed
in 1977, consistingof four single passmechanicallyaer-
ated aerationbasins(T2A and T2B) (Figure 4.49). These
basinswere designedto operateconventionally,by step-
feed, or completely mixed with separateintroduction of
RAS.
In 1982,when study of the Hamilton plant was initi-
ated(Wheeleret a1.,1984),the SVI of the T3 systemwas
usually in the range 80 to 150 ml/g. In the T2A and T2B
systems,SVIs below 200 mLlg were rarely encountered.
The bulking was largely due to Type 1851,N. limicolall,
Type 0675 , and Type 0041 filamentous organisms.In late
1982, the T2A and T2B systems were repiped so that
primary effluent and RAS flowed together in the step-feed
channel before entering the aeration basin. This channel
was aerated gently and it provided an averagehydraulic
 Control of ActivatedSludgeBulkingand Other SettlingProblems 121

To clarifier To clarifier

Ts,
To clarifier To clarifier
T6 T6

(al (b)

To clarifier - - - - - .- - .;. Infl uent - - - - - ,- - - > Effl uent

-----.-> RAS O Aerator
- CombinedinfluentanJ
----------->
R AS
To clarifier
T6

(c.)

FICURE 4.49 Modification of influent and RAS introduction points to aeration basins, Hamilton, OH: (a) original system, (b)
modification of both systemsto selectorconfigurationwith a hydraulic detentiontime of approximately4 min, and (c) modification
of systemT2A to provide a selectorwith a hydraulic detentiontime of approximately7 min. (From Wheeler,M. et al. (1984), Water
Sci.Technol.,16:10-11, 35. With permission.)

L*
L__
500

b0
) 4oo
Fi

a
300

200

100

0
1982 1983 1984 I 985 r987
Year

FICURE 4.50 Monthly averageSVI, Hamilton, OH, 1982 through 1989

retention time of approximately 7 min at averagesewage
flow. Figure 4.50 shows that, following selectorinstalla-
tion, the SVI decreasedgradually over an approximate
7-month period and by mid-1983 reachedthe same SVI
rangeexperiencedby the T3 system.Over the ensuing6 y,
two noteworthy events took place.
First, during 1985, the T3 system was taken out of
servicefor rehabilitation.For about 6 mo, the entire sewage
flow was treatedin the T2A and T2B systems.Figure 4.50
(and in more detail Figure 4.51) show an increasein SVI
during this period suggestingthat the selector(now with
an averagehydraulic retentiontime of approximately5 min)
was overloaded and soluble organic matter was breaking
through into the main aeration basin. Once the T3 system
was returned to service,the SVI values gradually returned
to their previous levels. This suggeststhat the T2 system
selectorwas sized very close to the limit for providing
effective soluble organic matter removal, and may also
explain the long period initially required for it to reduce
SVI.
122 Manualon Causesand Control of ActivatedSIudgeBulking,Foaming,and Other SolidsSeparationProblems

c. Davenport, IA
This casehistory illustrates the use of aerobic and anoxic
n rR selectors.Albertson (1990) reported results of 2.5 y of
work on the use of aerobic selectors for bulking sludge
f1 6 control at Davenport. The plant has an aeration basin
consisting of eight completely mixed cells aeratedby tur-
bines. The aeration cells can operate as a completely
mixed system with influent wastewaterfed to each aera-
tion cell; it can also operate in a seriesmode with two to
8 eight aerationcells in series.
250
Even though the plant was operated in series mode
200 with eight compartments in series, sludge bulking inci-
b0 dents due to Types 0675,1701, and 0041 occurred and
! rso the SVI reachedvalues in excessof 300 ml/g. Nocardio-
ri
form organisms were also present. The bulking sludge
K 1oo limited the plant peak flow to 20 to 25 MGD (0.88 to
F
1.1 m3/s) - far below the design value of 40 MGD
50
(1.8 m3ls).In late 1987, the plant staff startedto control
DO levels in the first aeration stage (the initial contact
Apr Jun Aug Oct Dec Feb Apr Jun zone or ICZ) while operating the aeration basin in two
I 985 19 8 6 trains of three compartments each. When the ICZ was
Time operated with a very low aeration rate to produce a DO
concentrationof close to zero, the SVI decreasedfrom
FICURE4.51 Effectof influentflow on T2 systemSVI,Hamil-
over 300 ml-/g to less than 100 mllg over a l-month
ton,OH.
period (Figure 4.52). Further studiesin 1988 showedthat
The secondevent was the failure in May 1985 of the reinstating full aeration inthelCZ induced bulking again.
air compressorused to generateair for selectoraeration. When the ICZ was operatedwith no aeration, an SVI
It was not replaced. Since the Hamilton plant nitrified of 40 to 50 ml-/g resulted and led to dispersed floc and
completely, the selector became an anoxic rather than elevatedsecondaryeffluent SS concentrations.In 1989,
aerobic selector.From Figure 4.50, it appearsto function the practice of aerating the ICZ for 4 to 12 hl d to maintain
anoxically as well as (or betterthan) when it was aerated. the SVI to between80 and 100 mllg was adopted.When
The additional metabolic selectionprovided by the anoxic the SVI fell below 80 ml/g, the ICZ aeration period was
conditionsmay explain this. increased;when it exceeded 100 ml/g, theICZ aeration

360

320

280

^" 240
a 200
Fi

,h 160

t20

80

40

0
o 7o l4o 28o 3so
mn ot u#

IA plant.(FromAlbertson,
FIGURE4.52 BulkingcontrolatDavenport, O.E.(1990),WaterSci.Technol.,23:44,835.
With permission.)
Control of ActivatedSludgeBulkingand Other SettlingProblems 123

Mixed liquor recycle

Primary
effluent OO OO OO OO OO To
.O . o oO o o rO r o
.O . o rO o o
Retum .o o
secondary
vo o - vo vo vo
activated l.tn l .tn l .'n clarifiers
r o
l^
r o ' .'^ l ^ I
o l^
ror.i l ^ r o
l^
sludge ao
o- - oo ^ 0 o- " oo ^ o -o o ^ O el oo
;o % o o

Submerged
o o"
-----7r-- -*-o- ---o- o o" 'o ot
-----n- ----7T-
o-

mlxer
Anoxlc
selector

FIGURE 4.53 Configuration of an anoxic selector system at the Tri-City plant, Clackamas County, OR. (From Daigger, G.T. and
Nicholson, G.A. (1990), Res.J. Water Pollut. Control Fed., 62,676. With permission.)

period was decreased.Using this mode of operation,the
plant was able to operatewith peak flows in excessof
40 MGD (1.8 m3/s)and producea secondaryeffluentwith
cBOD5 of 7.4 mgtL and SS of 13.5mg/L (20-monthaver-
age values).The (F/IM),was typically in the range0.64to
1.2 kg BODr/kg MLSS/d.
d. Tri-City, Clackamas County, OR
This casehistory illustratesthe use of an anoxic selector.
The Tri-City operation is a 13.5-MGD (0.59 m3/s)
advancedsecondarywastewatertreatmentplant consisting
of preliminary treatment, primary treatment, activated
sludge, and effluent chlorination. Typical secondarytreat-
ment (effluent BOD, and TSS both <30 mg/L) is provided
during the winter, while advanced secondary treatment
(effluentBOD, andTSSboth <20 mg/L) is providedduring
the summer.Becausethe plant nitrifies during the summer
and influent total alkalinity is low, an anoxic selector was
incorporatedinto the aerationbasin to reducetotal alkalin-
ity consumptionand providebulking control (Figure4.53).
Diffused air aeration is provided throughout the basin.
The initial 20Voof the basin volume (selector) is baffled
from the rest and contains both mechanical mixing and
diffused aeration equipment. Anoxic conditions are pro-
vided by operating the mixing equipment (but not the
aeration equipment) in the initial zone and returning nitri-
fied mixed liquor from the effluent end of the aeration
basin.An internal mixed liquor recyile pumping capacity
of l00%oof the designdry weatherflow is provided.Fully
aerobic treatment is provided by turning off the mixer and
providing diffused air to the selector.
The datareportedhere are for anoxic selectoroperation
over three dry seasons(June through October); an average
SVI of 79 mL/g was achieved.Average selectorhydraulic
retention time was 86 min and selectorF/lvI (kg BODr/kg
MLSS/d) was 0.72.
The effectiveness of the selector was confirmed by
operating the aeration basin in both the anoxic selector
and fully aerobicstep-feedmodes.Figure 4.54 showsthat

flow anoxic flow anoxic flow anoxic

--R

Jun Aug Oct Dec Feb Ap. Jun Aug Oct Dec Feb Apr Jun Aug Oct
1986 1987 1988
Month and yed

FIGURE 4.54 Performance of an anoxic selector at the Tri-City plant, Clackamas County, OR (From Daigger, G.T. and Nicholson,
G.A. (1990), Res.J. Water Pollut. Control Fed., 62,676. With permission.)
124 M anua lo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
the SVI increased whenever the aerobic step-feed mode
was used and decreasedwheneverthe anoxic selectorwas
used. Aeration of the selector, practiced in August and
September1987,alsoincreasedthe SVI. The anoxic selec-
tor at Tri-City has sometimesbeen too effective. On occa-
sion it has reduced the SVI as low as 20 to 30 ml-/g and
a turbid effluent resulted. This was countered by periodi-
cally providing a low level of aeration in the anoxic zone
to encourage the growth of some low DO filamentous
organismsand increasethe SVI.
e. Fayetteville, AR
This casehistory illustratesthe use of an anaerobicselec-
tor. The current Fayetteville plant commenced operation
in March 1988 and replacedan existing activatedsludge
systemwith a completelymixed aerationbasin that had a
long history of producing bulking sludge. The current
facility was designedto nitrify and provide EBPR (sup-
plemented by alum addition) and also provide bulking
sludgecontrol.The plant treatsa mixture of domesticand
food processingwastewaters.
The aeration basin is configured so that it can be
oper at ed in b o th a n a e ro b i c /a e ro b i c a n d anaero-
bic/anoxiclaerobic modes. Operation in the anaero-
bic/anoxic/aerobicmode is accomplishedby using the
mixed liquor recyclepumps.The anaerobic/aerobic
is typically used.In this mode,the anaerobicselectorcon-
sists of six completely mixed cells in series,followed by
an aerationbasinconsistingof four completelymixed cells
in serieswith mechanicalsurfaceaerators(Figure4.55).
Mixed liquor recyclecapabilitywas also providedbut was
not used during the evaluationperiod reportedhere.
From June I 988 throughAugust 1989, the Fayetteville
plant operated with an initial selector zone hydraulic
residencetime of 15 min and an initial selector zone
organic loading of 1.6 kg BOD./kg MLSS/d. Total aera-
tion basin hydraulic residencetime was t h and F/M was
0.045 kg BOD,/kg MLSS/d.
Since start-up of the anaerobicselector system,the
SVI at Fayetteville has averaged 86 mLlg and has not
exceeded185 mllg. With the plant configurationprior to
anaerobic selector installation, chlorination was used
almostcontinuouslyto maintainthe SVI below 150 ml-/g.
The highest SVI values were observedduring periods of
rainfall when dilute, highly oxygenatedwastewatercaused
the biological system influent to have a DO of 8 to l0
mg/L. Under these conditions, low DO filaments appar-
ently grow in the anaerobic selector.
f. Hyperion Treatment Plant, Los Angeles, CA
This casehistory describesthe useof an anaerobicselector
(Cheng,2002). Nine high purity oxygen activatedsludge
modules were constructedat the Hyperion treatment plant
to upgradethe existing primary and partial secondaryplant
to full secondary treatment with a total capacity of
Pr oblem s
Anaerobic
selector
Primary
effluent
Return
activated To
sludge secondary
clarifiers
FICURE4.55 Configurationof anaerobicselectoractivated
sludgesystemat Fayetteville,
AR.
mode
450 MGD (19.6 m3/s).Each moduleconsistsof threeacti-
vated sludge trains and four secondaryclarifiers. Each
activatedsludge train consists of four cells, the first of
which is 54 ft (16.4 m) wide, 108 ft (32.9 m) long, and
30 ft (9.1 m) deepwith a 25 ft (7.6 m) waterdepth.This
cell had two 125-hp(93.5 kW) draft tube surfaceaerators.
Each of the following three cells is 54 ft (16.4 m) wide
and long, 30 ft (9.1 m) deep with a 25 tt(7.6 m) water
depth and is equippedwith a 7.5-hp (56 kW) draft tube
surface aerator.
Immediately following start-up, significant foaming
problems due to nocardioform organismsarose.To combat
this problem, the MCRT was lowered and SVIs well in
excess of 200 ml-/g resulted. Because of this, it was
decidedto test the possibility of operatingthe plant as an
anaerobic selector system to allow the control of both
nocardioform organismsand the filamentous organisms
that causedpoor settling.Two of the nine modules were
modified to anaerobic selector operation by dividing the
first cell into two compartmentsof equal size by construct-
ing a wall between the two halves. The draft tube aerator
was removed from the first compartment and replaced
with a 25-hp (18.7 kW) submergedmixer. The result was
a stagedsystemconsistingof five equal sizedcells- the
first mixed and the remaining four aerated.The two mod-
ified modules were operatedin parallel with the existing.
unmodified modules to determine the effectivenessof the
modifications in improving settleabilty.
Cont r olof A c t iv a te dS l u d g eB u l k i n ga n d Oth e r S ettl i ngP robl ems
150
b0
..1
-i
Z roo
50
May Jun Jul
FIGURE 4.56 Effect of an anaerobicselectoron the SVI of the Hyperion oxygen activatedsludgeplant in Los Angeles,CA. (From
Cheng, P. (2002), personalcommunication.With permission.)
Figure 4.56 presentsa comparison of the activated
sludgesettleabilityin the unmodifiedmodule(control)and
the modified module (selector).The operatingconditions
for both modules were as follows: averageinfluent flow =
53 to 55 MGD (2.3 to 2.4 m3ls);SRT = 1.5 d; MLSS =
1200to 1350mg/L; mixed liquor temperature>20nC,and
F/M = approximately0.9 kg BODr/kg MLSS/d. Neither
module nitrified. The F/M in the initial aeratedcompart-
ment of the control system was approximately 2.4 kg
BODr/kg MLSS/d, while in the unaeratedselectorsystem
compartment it was approximately 4.3 kg BODr/kg
MLSS/d. Figure 4.56 demonstratesthat the increasein
(F/tr4),and the change from aerobic to anaerobic condi-
tions in the first compartmentresulted in a decreasein
SVI from 2200 mL/g to about 100 ml/g.
g. 23rd Avenue Plant, Phoenix, AZ
This casestudy is discussedbecauseit illustratesthe abil-
ity to convert a high rate, nonnitrifying activated sludge
plant with bulking sludge to a nitrification-denitrification
plant producing effluent with a total inorganic nitrogen of
<7 mg/I,, without additional aeration basin volume or air
supply (Albertson and Hendricks, 1992).
The 23rd Avenueplant was designedto handle37 MGD
(1.6 m3/s), and was operated for many years at very low
MCRT (0.8 to 1.3 d) and thereforevery low MLSS con-
centrations (400 to 600 mg/L) to avoid nocardioform
organism growth, foaming, and partial nitrification. This
mode of operation resulted in severe bulking sludge
(SVI = 300 to 600 ml/g) causedtypically by the low DO
125
Aug Sept Oct Nov Dec Jan
Month,2000-2001
Type 1701,S. natans,andH. hydrossisfilamentousorgan-
isms and also by Thiothrix spp. and Type 021N. High rate
operationalso causedthe oxygen transfercoefficient(a)
to be very low; valueson the order of 0.21 were estimated
and O, transferefficiency was approximately6.97o.
High rate operation was conducted in the four-pass
aerationbasinsby stepfeeding primary effluent to PassesI
and 2 becauseof oxygentransferlimitations(Figure4.57a).
In this mode,the plant could treat about 24MGD ( I .0 m3/s).
In late 1989 through early 1990, the plant was con-
verted by operations staff to a nitrification-denitrification
plant incorporatingan anoxic/aerobicselectorsystem.The
plant was configured as a four-passaerationbasin with
the RAS and primary effluentfed to the headend of the first
pass.Mixed liquor was recycledfrom the fourth passof the
aerationbasin to the head end of the first pass.The first and
second passeswere compartmentalized(Figure 4.57b) to
form a five-compartment anoxic/aerobic selector system.
The first three selector compartments were mixed by
coarse bubble aeration; the fourth was provided with a
combination of medium and coarse bubble aeration to
allow either anoxic or aerobic operation; the existing fine
bubble diffusers were relocated from the first oass to the
secondpass.
Figure 4.58 shows the dramatic improvement in oper-
ation and performance that occurred following selector
systeminstallation. The SVI decreasedfrom routine values
of about 300 ml-/g (often exceeding600 mL/g) to approx-
imately 80 to 120 mllg. This improvement allowed the
126 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Pr oblem s

To secondary
clarihers 1680FB

1680FB
Internal
mixed
l 820FB liquor
Primary
effluent
2 1 0 0F B RAS OX 1A X 4
I C B /610M
Primary ttl
effluent rzort rzort i
l :ortl oort I I
(a) (b)
AX = Anoxic
OX = Oxic
FB = Find bubble
MB = Medium bubble
CB = Coase bubble

FIGURE 4.57 Modification of aerationbasin to an anoxic selectorconfigurationat the 23rd Avenue wastewatertreatmentplant,
Phoenix,AZ. (From Albertson,O.E. and Hendricks,P. (1992), Water Sci. Technol.,26, 461. With permission.)

Selectorsystem will increase{lt values,typically to a range of 0.4 to 0.6.

installed Combinedwith this effect, an anoxic selectorprovidesan
environment where soluble organic matter is removed in
l t0 the absenceof aeration.Therefore, the zone that contains
Air use m3/kg BoD
90 high soluble organic matter concentrations(which reduce
oxygen transfer efficiency) does not require aeration.The
1i
combined effect of these factors allowed the plant to treat
50 a given BOD, load with the same amount of air (m3 air)
Flow L/s x 102
SVI x l0r ml-/g
in ths nitrification-denitrification (selector) mode and
achieve total inorganic nitrogen removal to <7 mg/L sup-
plied/kg BOD, removed that was required in the high rate
non-nitrogenremoval mode (Figure 4.58).
In the nitrification-denitrification anoxic selector
RAS, SS x 103mg/L mode, the fine bubble diffuser oxygen transfer efficiency
MLSS mg/L x 102
increasedto l27o with an averagecr value of 0.47; in the
last two passesthe c, value was 0.52.
tiirl"r run sepNov ti;#* JunSepNov
'"" '- "" "':ny,"t 6. Situations Where Selectors Are Not Effective

Filamentousbulking problems that are not amenableto

FIGURE4.58 Effectot r"r".,i, ,rrtem on SVI, MLSS,RAS
treatmentplant,
SS,and air useat the 23rdAvenuewastewater
Phoenix,AZ. (FromAlbertson,O.E.(1990),Water Sci.Technol.,
23:4-6,835.With permission.)
plant to operateat MLSS valueson the order of 3000 mg/L
(with RAS SS of 9000 mg/L or more). The plant could treat
wastewaterflows of approximately33 MGD (1.45m3/s)
and produced an effluent with averageconstituentconcen-
trations of cBOD, = 5.4 mg[,, SS = 6.6 mg/L, and total
inor ganic N= 5 .8 m g /L .
Converting to an anoxic/oxic selector system also
improved oxygen transfer efficiency by providing condi-
tions that increased the cx value. In a high rate activated
sludge system (MCRT <2 d), u values are suppressed,
most likely because of the presenceof surface-active
soluble organic matter in the mixed liquor. Reducing F/M
(increasingMCRT) to provide a nitrifying activatedsludge
resolution by the straightforward application of selectors
occur most often in activated sludge plants with aerobic
and anoxic zones that are operatedat high MCRT values
(above approximately 12 d). Since this type of plant
includes nitrification-denitrification. EBPR. and BNR
activatedsludge systemsthat are becoming more common
in the U.S., the bulking problems assumegreat impor-
tance, particularly the growth of the M. panicella fila-
mentous organism that can lead to increasesin SVI and
foaming problems. The current knowledge of the causes
and controls for M. parvicella growth and the resulting
bulking and foaming are presentedin Chapter 5.
Other filamentous organisms that appear in high
MCRT systemsincorporating anaerobicand anoxic zones
and are not significantly affected by selectors are Types
0041,0615, and 0092. Types0041 and 0675 do not often
grow to levels that causebulking problems. Eikelboom et
al. (1998) associatedincreasedlevels of Type 0041 with
Cont r olof A c ti v a te dSl u d g eB u l k i n ga n d Oth er S ettl i ngP robl ems
plants that have initial anaerobic selectorsin which RAS
and raw (unsettled) wastewatermix followed by an acti-
vated sludge system that provides alternating anoxic and
oxic conditions, e.g., the Danish Bio-Denitro activated
sludge modification.
Type 0092 can causesevereepisodesof bulking. This
filamentous organism grows at high MCRT in activated
sludge systemsthat have anoxic zones,and especiallyif
the anoxic zones alternatewith aerobic zones as in an
oxidation ditch. Gabb et al. (1990) showedthat a contin-
uously fed, intermittently aerated laboratory activated
sludge system (simulating an oxidation ditch) and oper-
ated at an MCRT of 20 to 30 d bulked severely due to the
proliferation of Type 0092 and M. parvicella. The instal-
lation of an aerobic selector ahead of the intermittently
aeratedbasin did not eliminate these filamentousorgan-
isms. When the aerationbasin was continuouslyaerated,
bulking due to both Type 0092 and M. parvicella was
cured. These results suggestthat Type 0092 is a slow
growing filamentous organism that gains a competitive
advantageunder nitrifying-denitrifying conditions.
The following ideas have been proposedas possible
reasonsselectorsare unable to control this group of fila-
mentousorganisms:
. The filamentous organismshave high substrate
uptake and storagecapacity (possibly M.parvi-
cella).
. The filamentousorganismsgrow on eithercom-
plex organic matter or particulate material
whose concentrationis not influenced in a
selectorsystem (possiblyTypes 0675, 0041,
and 0092).
. The filamentousorganismsdenitrify (possibly
M. parvicella and Type 0092).
Table 4.8 shows that the denitrification rate of an
activatedsludsedominatedbv M. parvicella is of the same
TABLE4.8
Denitrification Ratesfor Type021N, G. amarae
Strains,Z. ramigera,and an ActivatedSludge
Dominatedby M. parvicella
DenitrificationRate
Strain Typeof Oganism (mg NOr-N/g SS/min)
Type 02lN Filament (bulking) 8.3x l0r
G. amarcte ASF3 Filament (foaming) 3.2x 10r
G. amarcte ASACI Filament (foaming) 2.4 x 10:r
Z. ramigera Selector floc former 3.3x10 I
M. pawicella- Filament (bulking 1. 8t o 2 . 7x l 0 r
dominated and foaming)
activated sludge
127
order of magnitude as the selector floc-former Z. ramig-
era. Because most of these filamentous organisms grow
inside the flocs (producing diffuse floc structures), their
control by RAS chlorination can require high and pro-
longed chlorine doses that can cause some deterioration
in effluent quality and compromise EBPR. Experience
indicatesthat the following approachescan help to control
the growth of these filaments:
. Stagethe anaerobic,anoxic, and aerobiczones.
Staging produces substrate concentration gra-
dients and allows kinetic selectionto exert its
effect in controlling the growth of these slow-
growing microorganisms.
. Operate at the minimum possible aerobic
MCRT. Experienceindicatesthat the growth of
theseorganismsis encouragedby operationat
higher aerobic MCRTs than necessaryfor
achievingprocessobjectives(Marten and Daig-
ger, 1997).Apparently the higher than neces-
sary MC R T al l ow s these sl ow growing
microorganismsto becomeestablished.
. Avoid low DO concentrationsin the aerobic
zone. It appearsthat some of these organisms
are ableto grow well at low DO concentrations,
and especially when the DO concentrations
cycle betweenlow and high values.A DO con-
centrationof at least 2 mg/L should be main-
tainedconsistentlythroughoutthe aerobiczone.
Special attention should be paid to providing
sufficient aeration capacity to maintain DO
where the selectorcontentsflow into the main
aeration basin. The DO uptake rates can be
quite high at this point (up to approximately
60 mg Orlg SS/h).
V . HI G H E F F T UE NT
S SDUET O
CLARIFICATION PROBTEMS
A. Grrurnnl
Earlier in this chapter,it was noted that secondaryclari-
fiers provide the functionsof clarificationand thickening.
Thickening failure can produce elevated effluent SS
becausethe clarifier lills with sludge,causingthe sludge
blanket to spill into the effluent.In contrast,clarification
failure can result in elevatedeffluent SS concentrations
when the sludge blanket is low. Clarification failure is
relatedto the inability of the clarifier to settleand remove
the clarifier influent SS.
The clarification function is affected by many factors,
including clarifier hydraulic loading and configuration,
density currents, short-circuiting etc. (Ekama et al., 1997;
Clarifier Design, 1985).Elevatedeffiuent SS also can occur
if the mixed liquor enteringthe secondaryclarifier is poorly
128 M anualo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
flocculatedandcontainsparticles that are too small to settle
in the clarifier. Poor flocculation can result when the mixed
liquor is poorly flocculated in the aeration basin or
becomes poorly flocculated (floc break-up) as it is con-
veyed to the clarifier from the aeration basin (Das et al.,
1993). In some cases,poorly flocculatedsludge can be
flocculated prior to settling in a secondary clarifier to
reduceits dispersedSS content and therebyreduceeffluent
SS concentrations(Wahlberg et al., 1994).
B. PnosLrA
DrrrurrroN
1. Methodof Investigation
Procedures(describedin Chapter2)have been developed
to determinewhether effluent SS is a result of biofloccu-
lation failure, floc break-up, or secondaryclarifier hydrau-
lic problems.(Wahlberg,2001; Wahlberget al., 1995).A
dispersedSS test can be performed at severallocations
including the secondaryclarifier inlet (DSS'), the liquid
leaving the clarifier center well, and the clarifier effluent
weir (DSS").A flocculatedSS testis performedon a mixed
liquor sample(FSS).An effluent SS analysisis made on
a secondaryclarifier effluent sample (ESS) taken at the
sametime as the samplesfor DSS" measurement.
2. Results
The data obtainedby the above methodscan be used to
diagnosethe causesof poor secondaryclarifier perfor-
mance(i.e.,high ESS) as follows:
. High ESS, high DSS,.low FSS - The mixed
liquor doesnot haveadequatetime to flocculate
after it leavesthe aerationbasin or some con-
veyance structure between the aeration basin
and secondaryclarifier (e.g.,pump, free fall, or
tortuouspiping network) is causingfloc disrup-
tion. The DSS test can be repeatedat several
placesin the aerationbasin,in the mixed liquor
flow scheme.and at the effluent weir to locate
the sourcesoffloc disruption.This type ofresult
suggeststhat the ESS can be loweredby elim-
inating floc breakup and/or by incorporating
reflocculationprior to settling.
. High ESS, low DSS' low FSS - The mixed
liquor entering the secondary clarifier is well
flocculated and the high ESS are most likely
due to hydraulic problems in the secondary
clarifier. If the ESS value is higher than the
DSS, value and the sludge blanket is not high,
scouring of the sludge blanket is indicated.
. High ESS, high DSS,,high FSS- This result
indicates a bioflocculation problem leading to
dispersedgrowth. Bioflocculation problems can
P roblem s
arise from a variety of causes such as low
MCRT or the presenceof a dispersant,surfac-
tant. toxicant. or unfavorable monovalent:diva-
lent cation ratio in the wastewater.
. High ESS, intermediateDSS,,low FSS-This
result indicates both bioflocculation and
hydraulic problems.
3. Problem Resolution
a. Inadequate Flocculation, Floc Break-Up
(High ESS,High DSS,, Low FSS)
Mixed liquor can havehigh DSS,and low FSS becauseof
inadequatetime or conditions for flocculation or from the
break-up of already-formed flocs by shearing.Flocs con-
taining no filaments or with very low filamentousorganism
levels can be especially prone to theseproblems resulting
in pin floc.
Sludgeflocculationcan be accomplishedby reducing
the mixed liquor energyinput from a level that causesfloc
shear(G > 125s-r;Das,l993)to onethat allowsthe sludge
particlesto aggregate(25 < G < 100 sr). Shearlevelscan
be reducedin a diffused air systemby reducingthe aera-
tion rate at the effluent end of the aeration basin (i.e..
taperedaeration).
Figure 4.59 shows the effect of energy input from
diffused air aerationon the DSS level in activatedsludge.
For activatedsludge systemswith mechanicalaeration,
the mixed liquor discharge to the secondaryclarifiers
should be locatedas far as possiblefrom an aeratorsince
these devices produce high localized velocity gradients.
Large hydraulic drops, mixed liquor pumping, and tortu-
ous piping/valvingduring mixed liquor conveyanceto the
secondaryclarifier shouldbe avoided.Shouldany ofthese
conditions be unavoidable,a properly designedfloccula-
tion zone should be installed.
J
;30
E
6
a)n
F
610
(h
0
r00 150 200 250 300
0 50
G, s-l
FICURE 4.59 Variation of supernatant suspendedsolids con-
centration with mean velocity gradient (G) for diffused air acti-
vated sludgeplants.(From Das, D. et al. (1993), WaterEnviron.
Res.,65,138.With permission.)
Cont r olof A c t i v a te dS l u d g eB u l k i n ga n d Oth e r S ettl i ngP robl ems
TABTE4.9
DSS,FSS,and ESSConcentrationsbefore and after
Clarifier Modificationsat Creeley,CO (mg/t)
Clarifierlnlet Effluent
Clarifier Condition DSS(DSS') Weir DSS FSS
Before modifications ,a 22 8.2
After modifications t7 5.3 7.0
Source: From Brischke, K. et al. (1997), Proc. Water Environ. Fed. 70th
Ann. Conf., p. 237. With permission.
Such a zone, with an averagehydraulic retention time
of about 20 min, can be provided within the secondary
clarifier. No mechanical mixing is required in this floccu-
lation zone because the energy resulting from the dissi-
pating momentum of the influent flow is sufficient to cause
flocculation(Parkerand Stenquist,1986).In circular sec-
ondaryclarifiers,sucha zonecan be providedby enlarging
the centerwell and designingfor mixing using a tangential
mixed liquor input. Further discussion of this topic
appears elsewhere (Grady et a1., 1999; Wahlberg et al.,
1994; Das et al., 1993; Ekama et al., 1997).
Parker et al. (2000) report on the use of the Wahlberg
(1995) techniquesto investigateand rectify a problem of
inadequateflocculationat the Greeley,CO plant. DSS/FSS
testing of the existing secondary clarifiers (Table4.9)
showed poorly flocculated clarifier influent solids and
inadequateflocculation in the clarifier. After the clarifiers
were modified by converting the inlet structure to a floc-
culator centerwell (Parkeret al.,1966), the effluent weir
DSS (5.3 mg/L) and ESS (6.3 mg/L) valueswere closeto
the FSS value(7.0 mg/L;Table 4.9).
b. Clarifier Hydraulic Problems (High ESS,
Low DSS, Low FSS)
In this condition,the mixed liquor entersthe clarifier with
low DSS; its low FSS level indicatesno difficulties with
bioflocculation,but the ESS valuesare high. Theseobser-
vationsmean that some physical or mechanicalcondition
in the clarifier produces high ESS from activated sludge
that should produce a low ESS.
A detaileddiscussionof secondaryclarifier mechan-
ical and hydraulic problems and their resolution is beyond
the scope of this manual. Factors such as shape, depth,
weir placement, center well and inlet structure design,
sludge removal mechanisms, hydraulic and solids load-
ings, and baffling can be important. Comprehensivecov-
erageof thesetopicscan be found in Wahlberget al.,1995,
Ekama et al., 199"7;Daigger and Butz, 19981,Design of
MunicipalWastewater TreatmentPlants, 1998, and Wahl-
ber g, 2001.
129
TABLE4.10
Clarifier TestResultsfrom the
Central Marin Sanitation
Agencyin California (mg/L)
ESS Location DSS ESS
26 Center well, inlet l0
6.3 Center well, outlet il
Effluent weir 3.6 8.5
Soarce:From Parker,D.S. et al. (2000), Water
Sci.Technol.,4l:9, 201.With permission.
The results of an investigation on the secondaryclar-
ifiers at the Central Marin Sanitation Agency plant in
California illustrate a combination of flocculation and
cl ari fi er hydraul i c probl ems (P arker et al . , 2000;
Table4. l0). No FSS datawere compiledduring this inves-
tigation. The results show that the very small center well
did not provide for flocculation and deflocculation
occurred in the secondaryclarifier. ESS were significantly
higher than effluent weir DSS, showing that hydraulic
inadequaciesin the secondaryclarifier causedflocculated
and settleableTSS to passinto the effluent.
c. Bioflocculation Problems (High DSS,,
High FSS,High ESS)
High valuesof FSS,DSS,,and ESS suggestthat no matter
how good the flocculation and settling conditions are, a
sludge cannot produce a low ESS becausesomethingis
amisswith its ability to flocculate.This is the leastunder-
stood area of activated sludge SS separation,although
some advancesin our understandingof the fundamental
mechanismsof bioflocculationarebeginningto shedsome
light on it. Becauseof this poor understanding, a common
approach to dealing with poor bioflocculation is to add a
chemicalflocculatingagent(e.g.,the salt of a hydrolyzing
metal ion such as Fe3+,Al3+,or an organic polymer) to take
the place of biologically producedflocculatingagents.
Causesof dispersedgrowth include the presencein
the wastewaterof poorly biodegradableor nonbiodegrad-
able surfactantsand dispersants.toxic compoundsin the
influent, wastewaterswith high monovalentcation (Na*,
K*, NHl*) to divalent cation (Ca2*,Mg2*) ratios, activated
sludge with a low MCRT (Figure 4.60), high aeration
basin temperature, and overchlorination of activated
sludge during bulking control.
Murthy et al. (1998) reported the effects of adjusting
the monovalent:divalentcation ratio of an industrial waste-
water on the settleability and dewatering characteristics
of activated sludge. Poor settling occurred in the absence
of filamentous organisms and was accompaniedby copi-
ous dispersed growth. The major component of the high
130 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

20 1000

3 16
Fi
18 o BisogniandLawrence
o Choa andKeinath
(1971) E aoo
-l

i
t-*'-
MgSOa r
b 14 E 600 addition rtr
s12 r
a begins
ui
(^ 10 a E 400 .J-
EU a -.L .-
@
9K a I zoo
o
O/ 6l
'a a
E2 0
a
80
0
02468r 0 1 2 Time, d

Mean cell residencetime, d

FICURE4.61 Effect of MgSOoadditionon 2-hour activated
timeon dispersed
FICURE4.60 Effectof meancellresidence sludge settledvolumeduringfield trials. (FromMurthy,S.N.et
al. (1998),WaterSci.Technol.,38:3, 119.With permission.)
suspended
solidsin activated
sludge.

strength(10,000mg COD/L) wastewater was2000mg/L sludge settling occurredwithin 2 wk (Figurc4.6l). Murthy

of aceticacid.To preventvolatilizationof the aceticacid,
the influentpH was adjustedwith sodiumhydroxideso
thatthemixedliquor pH was8.8.Duringfield trials,some
of the NaOH used for pH adjustmentwas replacedby
magnesiumsulfate(MgSO). Improvementsin activated
et al. (1998) suggest that the use of MgSOo in place of
some NaOH may be a more economical method of pro-
ducing well settling activated sludge than using organic
polymers to improve the activatedsludge solids separation
characteristics.
 5 ActivatedSludgeFoaming
\''
and Control
They row'd her in across the rolling foam,
The cruel crawling foam,
The cruel hungry foam,
To her grave besidethe sea.
But still the boatmenhear her call the cattle home,
Across the sandsof Dee.
Charles Kingsley, 1819-1875, "The Sands of Dee"
I. ACTIVATED
SLUDGEFOAMING
A. TYprsor AcrrvRrroSluocr Fonpr
The formation of foam or scum on the surfacesof activated
sludge aeration basins and secondaryclarifiers will be
discussedin this chapter. Their formation has been
ascribedto a variety of causesincluding:
. White, frothy foam is observed during the
start-up (usually after 3 to 4 d) of activated
sludge plants, possibly due to the presenceof
undegradedsurface-activeorganic matter or to
the growth of Type 1863 in the aerationbasin.
This type of foam usually disappearswhen the
processbecomesestablished.
. When poorly biodegradable,branched-chain,
alkyl benzene sulfonate-based,household
detergents were marketed, persistent, volumi-
nous, frothy, white foams were produced on
aerationbasins.Since the introduction of bio-
degradablehouseholddetergents,this type of
foam no longer appearsin domestic wastewater
treatment plants. Foaming due to the presence
of slowly biodegradablesurfactantsoccursduring
the treatment of some industrial and municipal
wastewatersespeciallywhen the wastewateris
cold. Often theseare nonionic surfactantsin the
alkyl phenol ethoxylate(APE) or nonyl phenol
ethoxylate(NPE) families of compounds.
. A sticky, viscous foam may appear when acti-
vated sludge is nutrient-limited, probably due
to the presence of surface-active exocellular
polymeric material formed by activated sludge
microorganisms.Microscopic examinationusing
India ink stain will reveal poor floc penetration
by the carbon black particlesof the ink.
. Foaming or scum formation along with produc-
tion of a turbid effluentmay result from excessive
recycling of flne solids from anaerobically
digestedsludgesolids handlingprocesses(e.g.,
anaerobicdigestersupernatant, vacuumfilter or
belt pressfiltrate, and centrifugecentrate).This
type of foam has a volcanic or pumice-like
appearance.Microscopic examinationwill reveal
that it contains large amounts of amorphous
inorganic and organic particles.
. Scum can be producedon secondaryclarifiers
or in the anoxic zones of nitrifying activated
sludge plants by denitrification.Small bubbles
of nitrogen gas attach to the activated sludge
flocs and carry them to the surface.This phe-
nomenon in secondary clarifiers is referred to
as sludgerising or blanket rising.
. A stable, usually brown, foam or scum seen
widely on activatedsludgeaerationbasinsand
secondaryclarifiers ("Milwaukee Mystery,"
1969; Wells and Garrett, l97l; Lechevalier,
1975;Pipes,1978b;Dhaliwal, 1979)has been
associatedwith largenumbersof actinomycetes
containingmycolic acidsin their cell walls. We
will refer to these organisms collectively as
nocardioforms or nocardioform organisms.
They are taxonomically diverse (Lechevalier,
1975;Lemmer and Kropenstedt,1984; Soddell
et al.,1992; Schuppleret al., 1995;Goodfellow
etal ., 1996) and contai n members of t he
Corynebacterium, D i etzia, Gordona,My cobac-
terium, Nocardia, Rhodococcus,Skermania,
and Tsukamurella genera. A similar type of
foam was observedin the presenceof Micro-
thrix parvicella, a filamentous actinomycete
distantly related to nocardioforms (Blackall
et al., 1994, 1996).
In previouseditionsof this text and in most literature,
this type of foam has been referred to as nocardia foam.
Sincemany other typesof organismscan causeit, we will
adopt the phrase nocardioform foams (Soddell et al.,
r998).
131
132
M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
I I . NO C A R D IO F OR M F O A M IN G
A. oF Pnoslrlas
ExrrrurAND SrcNrFtcANcE
1. Foaming Organism Surveys
Surveys of nocardioform organism incidence in activated
sludge were conductedby Richard et al. (1982), Strom
and Jenkins (1984), Blackbeard et al. (1986), Seviour
etal. (1994), and Goodfellow et al. (1996) in conjunction
with surveysof filamentousorganismsin bulking activated
sludge. Nocardioforms were some of the most commonly
observed filamentous organisms in activated sludge; they
were the dominant filamentous organisms in approxi-
mately 3OVaof U.S. samplesand approximately l4%oof
South African samples.In the Australian samples,they
were the third most common fllamentousorganisms.
Some surveys were directed specifically to foaming
activatedsludge or to nocardioform organism incidence
in activated sludge. Seviour et al. (1990) surveyed 129
activatedsludgeplants in Queensland,New SouthWales,
and Victoria,Australia and found that 66 of the plantshad
foam problemsat the time of the survey.The most com-
monly found filamentousorganismsconcentratedin the
foam over their levels in the mixed liquor were M. parvi-
cella, Gordona amaree, and Skermania pinensis (previ-
ously Nocardia pinensis - "pine-tree-like-organism"or
PTLO;Blackall et al., 1989;Figure5.1d).The foamsalso
contained other filamentous organisms such as Types
0092, 0914,0041, and 0675 but these were present at
concentrationsno greaterthan in the mixed liquors from
which they were formed.
Wanneret al. (1998) surveyedthe filamentousorgan-
ism population of the majority of the activated sludge
plants in the Czech Republic and found that M. parvicella
was one of the most dominant in mixed liquors and the
most common in foams. Eikelboom etal. (1998) found
that M. parvicella was by far the most important filamen-
tous organism in nutrient removal plants in a survey of
more than 200 such systems in Denmark, Germany,
Greece,and The Netherlands.Pujol et al. (1991) surveyed
some 6000 activated sludge plants in France and found
that 2O7oof all plants and 87Voof the extended aeration
plants had foaming problems at one time or another. Of
the 58 activatedsludgesexaminedmicroscopically,45Vo
contained M. parvicella as the dominant filamentous
organism.
Pitt andJenkins(1990)conducteda nationwidesurvey
of activated sludge foaming in the U.S. Of the 114
responses,75 (66Vo)indicated that they experiencedsome
type of foaming at one time or another. As part of this
survey, the nocardioform levels in foam and mixed liquor
samples were determined for the plants in terms of a
numerical scale rating based on the subjective scoring
Pr oblem s
describedin Table 2.l5.The averagenumerical scalerating
for plants indicating a foaming problem was 4.2 (common)
compared to 2.1 (few) for those not indicating foaming
problems. N. amarae (now G. amarae) was determined
by Lechevalier and Lechevalier (1974) to be the most
common actinomycete present in activated sludge foam
in the U.S. This organismhas not been reportedin envi-
ronments other than activated sludse.
2. Foaming Problems
a. Activated Sludge
Significantquantitiesof a nocardioformfoam can cause
severeoperatingproblems(Figure 5.2).In aerationbasins
with submerged effluent structures, foam tends to accu-
mulate in the basins.It can accumulateto such an extent
that it overflows the basin freeboardand coverswalkways,
handrails,and surroundingareas,creatinghazardous,slip-
pery conditionsand limiting accessto equipment.In cov-
ered aeration basins, nocardioform foams have been
known to accumulate to such a degree that the water
contentof the foam trappedin the basinexceedsthe avail-
able head for gravity flow of peak wastewater flows
throughthe aerationbasin.
Processcontrol becomesextremely difficult if a sig-
nificant fraction of the solids inventory is presentin the
foam trapped in the aerationbasin. In one plant, it was
estimated that the foam contained 40 to 457o of the total
activated sludge suspendedsolids inventory. Hao et al.
( 1988)estimatedthat for a 4.5 m deepaerationbasin with
an MLSS concentrationof 3000 mg/L and a 7.5 cm deep
foam containing 5VoTSS, some 2OVoof the total sludge
inventory would be in the foam.
When foam escapesfrom an aeration basin into a
secondaryclarifier, it may overwhelm surface scraping
devicesand enterthe secondaryeffluent,elevatingeffluent
TSS (and BOD') levels. In cold climates, nocardioform
foam can freeze on the secondaryclarifier surfaces,mak-
ing scum removal devicesinoperative.Some plants rou-
tinely remove secondaryclarifier surface scrapersprior to
winter freezing to prevent damage by frozen foam. In
warm climates, the accumulated foam rapidly becomes
odorous.
A survey of U.S. plants with anaerobicdigester foam-
ing problems by van Niekerk et al. (1987) showed that
many digester foaming incidents can be attributed to the
presenceof nocardioforms in the waste activated sludge
fed to the digester.In laboratory-scaleanaerobic digester
experiments,van Niekerk et al. also showedthat digesters
fed with primary sludge alone did not produce stable
foam; when waste activated sludge containing nocardio-
forms was digestedwith the primary sludge, a stablefoam
was produced.
7', \

Activated Sludge Foamingand Control 133

FICURE5.1 Phasecontrastmicrographsof nocardioformandM. pamicellafoams;(a) and(b) nocardioforms (possiblyG. amarae),

(c) and (d) S.pinensis,and (e) nd (f) M. panicella. (Originalmagnification100x,(a), (c), and (e); 1000x,(b), (d), and (0.) (See
color insertfollowing page70.)

b. Anaerobic Digesters Foam penetrationbetweenfloating coversand

Anaerobic digester foaming due to nocardioforms has
caused the following operating problems in wastewater
treatment plants:
. Blockage of gas mixing devices leading to
escapeof gas through water seals
. Inversion of digester solids profiles
. Gas binding of sludgerecirculation pumps lead-
ing to an inability to heat the digesters
. Entrapment in, and fouling, of gas collection
piping
digesterwalls leadingto overflowof foamonto
the floatingcoveror ontothe groundsunound-
ing the digester(Figure5.2)
Pressurization of digesterdue to blockedgas
piping and pressurerelief valves leading to
structuralfailure of fixed roof.
Tipping of floating coversleadingto damageto
gasmixing equipment
Destructionof sludgelagoon coversand gas
collectionequipment(Figure5.2)
134 P r oblem s
M anualo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on

FIGURE 5.2 Nocardioform foaming in activatedsludge:

(a) foam overflowingaerationbasins,(b) operatorswading
through foam on walkway between aeration basins,
(c) foam escapingfrom plant into roadway,(d) anaerobic
digester overflowing with foam, (e) secondary clarifier
covered with thick foam layer, and (f.) sludge lagoon cover
and sludge withdrawal equipment destroyed by foam.
ActivatedSludgeFoamingand Control
*\-water
\_l__/
+6daa66da66adDr
(a)
FIGURE 5.3 Air-in-water foams of varioustypes: (a) without surfactant,(b) surfactantaccumulatingat air-water interface,and (c)
hydrophobic solids accumulating at interface. Note that solid particles can bridge acrossthe air bubble and prevent drainage of water.
c. PathogenicNocardioforms
Some of the actinomycetesisolated from activated sludge
foams are opportunistic human pathogens (N. caviae,
N. brasiliensis, N. asteroides, and strains of Mycobacte-
rium; Lechevalier, 1975; and N. farcinica; Stratton et al.,
1996). Although no incidents of human infection with
nocardioforms from wastewater treatment plants have
been reported, the potential exists that aerosolsgenerated
by aeration systemscan dispersetheseorganismsto where
they come in contact with treatment plant workers or
inhabitants of areas surrounding wastewater treatment
plants. The significanceof this potential should not be
underestimated, especiallywith the increasingnumbersof
immunologically suppressedindividuals in the general
population.
B. Pnoposro Mtcnnxtst*,tsor Fo*rlntc
1. Introduction
Factors that affect nocardioform foaming can be grouped
into those that encouragethe growth of nocardioforms in
activated sludge and those that encourage foaming.
Although interrelated,the former are largely microbiolog-
ical and the latter are mainly physical/chemical.The foam-
causing factors will be addressedfirst.
2. Physical/ChemicalFactors
a. Nature of Foam
Some foams are two-phasedispersionsof gas (air) bubbles
in a liquid (water) or solid (Styrofoam). For air-in-water
foams, the air is the dispersed phase and water is the
continuousphase(Figure 5.3a). Foams collapsethrough
thinning of the water layer separatingthe air bubble from
the surrounding air to the point where the pressureinside
the bubble is great enough to rupture the liquid film and
burst the bubble. Liquid film thinning takesplace by drain-
135
,,,,,, ,-';ffi'"iy*:*
,ko6ddbd6aa6dd6
U;/
(b)
Hydrophobic particles
age (gravity flow) and evaporation.Foams can be stabi-
lized by surfaceactive agents(surfactants)- molecules
attracted to the air-water interface (Figure 5.3b). In a
sense,thesemoleculesstrengthenthe water fllm between
the air bubbles and allow it to become thinner before it
ruptures.Foamsgeneratedfrom solutionscontainingsur-
factants therefore last longer than those produced in the
absenceof surfactants.
b. Solids-ContainingFoams
Another way to stabilize foams is by the addition of hydro-
phobic particles (particles that are poorly wetted by
water). This producesa three-phasefoam consisting of
dispersedphasesof air, a hydrophobic solid, and a con-
tinuouswater phase.The hydrophobicsolid particlestend
to congregateat the air-water interface,again making it
stronger.If the hydrophobic panicles are large enough,
they may bridge the water film, effectivelycreatinga dam
that impedesliquid drainageand film thinning that even-
tually resultsin film rupture (Figure 5.3c).
It has been proposed that nocardioform organisms
have hydrophobiccell walls due to the presenceof long
chain mycolic acids on their surfaces(Minnikin, 1982).
Recent work suggeststhat other factors may also be
important in determining nocardioform cell-wall hydro-
phobicity (Stratton et al., 1998). If nocardioform organ-
isms grow in sufficientnumbersin activatedsludge,they
will render the flocs hydrophobic enough to attach to
air-water interfaces(e.g., bubbles in an aeration basin)
and be carried to the basin surface. At the surface, the
foam will drain and the scum or foam will becomemore
concentratedin terms of SS and nocardioform organism
content than the mixed liquor. SS concentrationsas high
as 4 to 6Vohavebeenobservedin collapsednocardioform
foams.
Pitt and Jenkins (1990), in their survey of U.S. acti-
vated sludge plants, found that aeration basin foams
always had higher numerical nocardioform organism
| 36 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
abundancethan the mixed liquors from which they were
derived. Wheeler and Rule (1980) reported that mixed
liquor contained up to 106 nocardioform microcolo-
nies/ml and that the foam generatedfrom it contained up
to 1012nocardioform microcolonies/ml. Vega-Rodriquez
(1983) observedthat when activated sludge containing
nocardioform organisms was aerated in the laboratory,
typically '75 to 9OVoof the nocardioform filaments (on a
filament length basis) were transferred into the foam.
c. Roles of Surfactants
The presenceof surfactantsenhancesthe amount of foam
producedby an activatedsludgecontaining nocardioforms
and increasesfoam stability. For surfactantsto exert this
effect, they must be poorly or slowly biodegradable so
that they persist in the aeration basin. Ho and Jenkins
(1991) demonstratedthis effect by conducting foaming
experiments on mixtures of various amounts of a slowly
biodegradablenonionic surfactant - an alkyl phenol
ethoxylateknown as Igepal C-620, using (l) an activated
sludge that contained nocardioform organisms from the
Victor Valley, CA activatedsludge system and (2) an acti-
vated sludge that did not contain nocardioform organisms
from the East Bay Municipal Utility District activated
sludgesystemin Oakland,CA (Figure5.4 and Figure5.5).
The Victor Valley sludge revealed considerabletrans-
port of SS from the activated sludge during foaming and
the foam height increasedsignificantly as the surfactant
concentrationincreased.A stable,brown, SS-containing
foam was produced.In the East Bay activatedsludge,the
SS contentdid not decreasesignificantlyduring foaming.
A small amount of very unstablewhite foam containing
little SS was produced.Foamingof activatedsludgecon-
taining nocardioform organismsreducedthe nocardioform
level in the activatedsludge almost three-fold.
Theseobservationssuggestthat a useful analogycan
be drawn between nocardioform and M. parvicella foam-
ing in activated sludge and the processof mineral benefi-
ciationby flotation(Willis, 1988).In mineralbeneficiation,
4000
J of surfactant could produce a modest amount of foam. A
u ,Jotal suspendedsolids before foaming test
__t __ _ _ _ _ _ _
= 3000
E
o o
E onset of severenocardioform foamine in the wastewater
B 2000
6 o Total suspendedsolids after foaming test
1000
01 23 45
mg/L
CTASconcentration,
FIGURE 5.4 Effect of foaming on mixed liquor suspendedsol-
ids containing various amounts of lgepal C-620 at Victor Valley,
Nocardioforms were present.
Pr oblem s
4000
J
Total suspendedsolids before foaming tesl
il
.; 3500
E
E :ooo
E
Total suspendedsolids after foaming test
d zsoo
E
2000
01234567
CTASconcentration,
mg/L
FIGURE5.5 Effectof foamingon mixedliquorsuspended sol-
ids containingvariousamountsof Igepal C-620 at EBMUD.
Nocardioformswere not present.(From Ho, C.F. and Jenkins,
D. (1991),WaterSci.Technol.,23:4-6,879.With permission.)
minerals of commercial value are separatedfrom nonva-
luable material (gangue)by flotation.A chemical(collec-
tor) is added to the ground-up mixture of mineral and
ganguein water.The collector specificallyadsorbson the
mineral particles and renders them hydrophobic. A sur-
factant is also added to enhance foam production upon
aeration.The hydrophobic mineral-collector combination
attachesto air bubbles and is floated to the surface. The
surfactantstabilizesthe float so that it can be skimmed off.
In'activated sludge, nocardioform and M. parvicella
filaments can be regarded as the hydrophobic collectors
for activatedsludge flocs. The aerationair producesa float
of activated sludge flocs stabilized by poorly degraded
influent surfactantsor biologically produced surfactants.
In this sense,it is betterto view nocardioformfoaming as
a flotation processrather than a foaming process.
In many instances of nocardioform foaming in acti-
vatedsludgeplants,anecdotalhistoryofthe foaming prob-
lem often revealsthat modestamountsof foam are present
much of the time and very severefoaming incidents with
rapid onsets(in a matter of hours) occasionallyoccur. It
is possible that the severe foaming events are related to
increasedsurfactant levels. Thus, one could reason that a
ROO nocardioform-containing activated sludge in the absence
.j
.:0 slug discharge of poorly biodegradable surfactant (e.g.,
700 : from clean-up processesin an industry) to this system
could provide foam stabilization that would causea rapid
€4
600; treatment plant.
o It is possible that surflactantsplay a biological role in
'5 addition to a physical/chemical one in stabilizing foam.
500< Bendt et al. (1989) showed that a pilot plant activated
sludge system operatedat a 6-d MCRT and 15 to 18oC
and fed with wastewatercontaining 30 to 150 mg lipids/L
containedvery few nocardioforms.When the influent lipid
concentration was increasedto 250 mg/L by adding veg-
etable oil, and 10 mg/L of a surfactant was added, the
 ActivatedSludgeFoamingand Control
Primary
sfflusnl RAS
YV
r)t
Aeration basin
*os *l J urou.n,
(a)
FIGURE 5.6 Details of aerationbasin outlets and secondaryclarifier overflows: (a) foam-trappingunit and (b) nontrappingunit.
(From Cha. D.K. et al. (1992), WaterEnviron. Res.,64,37. With permission.)
nocardioforms became the predominant microorganisms
in the activated sludge. Bendt et al. suggested that the
surfactantemulsified the lipids, making them more readily
accessibleas substratesfor nocardioform organisms.
d. Anaerobic Digester Foaming
One prerequisite for the production of foam or a float is
the presenceof a gas phase. Since all activatedsludge
systemsare aeratedin one way or another,it is not possible
to eliminate the gas phase from an aerationbasin. This
approach however is a viable method for reducing nocar-
L
dioform foaming in anaerobic digesters. All anaerobic
digesters produce gas from digestion, but a far greater
volume of gas is passedthrough the digesting sludgeby
gasmixing devicesthanby the gasproducedby digestion.
van Niekerk et al. (1987) showedthat the foam level
produced in laboratory anaerobic digestersfed a mixture
of primary sludge and waste activated sludge containing
nocardioform organisms was a function of the digester
mixing method. The most foam was produced by fine
bubble gas mixing followed by coarsebubble gas mixing
followed by mechanicalmixing (Figure 5.38).
These results suggestthat nocardioform foaming in
digesters may be minimized through the use of a mixing
system that does not introduce small- to medium-size
bubbles into the digester. Several anaerobicdigester mix-
ing methods can achieve this objective. External pump
mixing does not add any gas bubbles.
Anaerobic digester foaming due to nocardioforms can
occur prior to appearingon the aerationbasinsfrom which
the WAS is fed to the digester. This most likely occurs
because the TSS concentration in the digester is much
higher than in the aeration basin. Thus, a given volume of
anaerobic digester contents contains more nocardioform
filaments (and therefore more foam-producing hydropho-
bic surface) than the mixed liquor.
137
Primary
gfflugn1RAS
VV
)t
Aeration basin
nos*--l
(b)
e. Foam Trapping and Recycling
Nocardioform foam problems increaseenormously when
the floated material is trapped on the surfacesof treatment
units and especially when the trapped foam is removed
and recycled through the treatment plant. Foam trapping
occurs whenever the free water surface is intercepted by
a structure(e.g., a wall or baffle) that makes the water
leave the treatment unit by a subsurfaceroute rather than
by an overflow.
Cha et al. (1992) compared the nocardioform popula-
tions in laboratory activated sludge systems with the two
aerationbasin and secondaryclarifier configurationsillus-
trated in Figure 5.6. The trapping configuration had a
subsurfaceaeration basin draw-off and a scum baffle pre-
ceded the secondaryclarifier weir. The nontrapping con-
figuration had an overflow aeration basin outlet and no
secondaryclarifier scum baffle. Figure 5.7 shows that
nocardioform populations in the trapping unit were up to
five times higher than those in the nontrapping unit. This
nocardioform population increase was due only to foam
trapping since Cha and coworkers wasted both foam and
mixed liquor in the same proportions. The recycle of
wasted foam would increase nocardioform levels even
more.
The effect of foam trapping can be observed in full-
scale plants by comparing the amount of foam accumula-
tion on aeration basins with and without foam trapping
features. Figure 5.8 shows an aeration basin with three
passes.The first two passesare connected by subsurface
outlets, while the third pass has an overflow weir as an
outlet. Foam accumulateson the surface of the first two
passeswhile the third passretains very little, if any, foam.
One often hearsthe following questionposedconcern-
ing nocardioform foaming: Why is nocardia so common
now in the U.S. when it never was before? Perhapsthe
138 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

l6
IMCRT= 10d---------------- MCRT= 5 d!
(t)
l4
(n
,r ) Foam : : Foam
E oo12 - - --.-- -----rlr- -- foam
No - traooine -+
? s- tr a p p r n g : "- i
ov :
F x10
Ya

fr'5 s
Eg
o tr o

Rb a
i. e-
c2

0.01
120
Day of experiment

FIGURE 5.7 Effect of foam trapping on nocardioform organism populations in bench-scaleactivated sludge units. (From Cha, D.K.
et al. (1992), Water Environ. Res., 64, 37. With permission.)

Subsurface
port

Overflow
weir

To
Subsurface secondary
poft clarifier -

FIGURE 5.8 Effect of foam trapping outlets on surface accumulation of foam on passesof an aeration basin.

answerlies in the foam trapping and recycle issuediscussed Incineration (1.37o)

above. In modem activated sludge plants, it is common Gravitythickener(5.37o)
Other(12%)
practice to place scum baffles ahead of the weirs in
Landnll (2.
secondary clarifiers. Anything that floats in a secondary Beltfilter(1.3%
clarifier, (including nocardioform-rich activated sludge Headwo*s (27.1Vo\
Anaerobic digester (8.07o
particulates)is trapped by the scum baffles, removed, and
often recycled back through the treatmentplant. This recy-
cling of nocardioforms provides them with a significant
competitive advantage. In secondary clarifiers without
scum baffles, any floating material is washed over the (1.3%
Scumconcentrator
weirs into the effluent. In this type of secondaryclarifier, WAS wet well (6. Aerobicdigester( 167o)
floatation is disadvantageousfor microorganisms. RAS wet
Pitt and Jenkins (1990) in their survey of foaming well1o.3%o\

problems in activated sludge plants found that many of

the foam disposal methods recycled the material directly
or indirectly back through the plant. Figure 5.9 showsthat
65Vo of the methods used recycle the foam (i.e., to the
headworks, gravity thickeners, WAS wet well, RAS wet
well, aerobic digester).
An additional effect of foam trapping was suggested
by the results of pure culture chemostat experiments on
G. amarae strains isolated from San Francisco and Sac-
ramento activatedsludges(Blackall et at., 1991). The two
FIGURE 5.9 Methods of disposal of foam removed from treat-
ment basins. Methods represented by shaded areas can recyle
foam.(FromPitt,P.A.andJenkins,D. (1990),Res.J.WaterPollut.
ControlFed.,62, 143.With permission.)
chemostatconfigurations shown in Figure 5.10 produced
significantly different G. amarae growth forms. When the
chemostat culture effluent was drawn off by an overflow
weir, the G. amarae culture grew largely as clumps with
diameters of up to 500 mm suspendedin a relatively clear
t_ ActivatedSludgeFoamingand Control t 39

Feed

Subsurface
withdrawal
Aeration by surface 02 of culture
transfer

i---t"r
surface
withdrawal
of culture

Aeration by bubbling air

( a) (b)

FIGURE 5.10 Chemostatconfigurationsand their effects on G. amarae growth: (a) head-spaceaeration and subsurfaceculture
withdrawal producedispersedG. amaraegrowth and (b) bubble aerationand surfaceoverflow culture withdrawal produceclumped
G. amarae growth. (From Blackall, L.L. et al. (1991), Res.J. WaterPollut. Control Fed.,63,44. With permission.)

L-

liquid. When the chemostatculture withdrawal system
was changedto a subsurfacewithdrawaldevice(Koopman
r
et al., 1980), the G. amarae grew almost completely in
f -.-
the form of dispersedfilament fragments. Merely chang-
ing the configuration of effluent withdrawal allowed
switching back and forth betweenthesetwo greatly dif-
ferent culture morphologies.These observationscan be
explained by the fact that the hydrophobic G. amarae
filaments tend to float. With an overflow culture with-
drawal, floating filamentswill be selectivelyremoved.If
the filaments are able to aggregateand form clumps that
settlefasterthan they float, the clumps will be selectively
retainedin the chemostat.Hence, a clumped culture will
develop in a chemostat with a surface overflow culture
withdrawal.
When the culture withdrawal is subsurface,floating
f,laments will not be removed selectively.A dispersed
culture will be formed under thesecircumstancesbecause
growth in a dispersedform no longer offers the disadvan-
tage of loss from the culture by floating and, compared to
growth as clumps, dispersedcells have better accessto
the substrate.
These data can be applied to observations made in
activated sludge systems.When foam trapping occurs in
activated sludge, free-floating nocardioform fi laments are
usually seenin the bulk liquid outside the activatedsludge
flocs. Since free-floating nocardioform filaments expose
a greater amount of hydrophobic surface to the aqueous
phase than the filaments inside the flocs (nonfoam trap-
ping), they will provide greater enhancementof foaming.
Therefore, foam trapping increasesnocardioform levels in
activated sludge and produces a culture with more free-
floating nocardioform filaments and a greater propensity
to foam.
Recent work by Narayanan(2002) has shown that the
amount of foam produced by activated sludge can be
directly relatedto the concentrationoffree-floating nocar-
dioform filaments but does not bear any relationshipto
the floc-associated nocardioformfilament level. The rela-
tive abilitiesof floc-boundand free-floatingnocardioform
filamentsto producefoam is most likely the reasonShao
et al. (1997) were ableto eliminateactivatedsludgefoam-
ing by adding a cationic polymer at the Terminal Island
plant in Los Angeles.The polymer may act to flocculate
the free-floatingnocardioformfilamentsinto the activated
sludge flocs.
f. Foaming Tests
Caution must be paid in applying the resultsof laboratory
foaming teststo full-scale activatedsludge systems.In a
laboratoryfoaming test,the maximum depthof liquid used
is on the order of I to 2 ft. In an aerationbasin,the liquid
depth is usually 15 ft or more. In a foaming test, flocs
containing nocardioform organisms are brought to the
liquid surface.Becausethe length of the column of liquid
below a given areaof liquid surfaceis much greaterin a
full-scaleaerationbasin than in a laboratoryfoaming test,
it takes fewer nocardioform organismsto produce a stable
foam in a full-scale aeration basin than in a laboratory
foam test. Therefore, the typical laboratory foam test is
an insensitiveindicator of foaming in prototype aeration
basins(Figure5.11).
The previousdiscussionsof nocardioform foam forma-
tion and the influences on it of various physical/chemical
factors and reactor design indicate that foaming tests are
poor methods for estimating nocardioform populations in
activated sludge. Too many factors other than nocardio-
form population are involved in foaming. For this reason,
140 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

Foam test column Aeration basin actinomycetesin activatedsludge.Nocardioforms can use

a wide variety of organic substratesas carbon sources.
Theseinclude readily degradablecompoundssuchas sugars
and low molecular weight fatty acids and slowly biode-
gradable, high molecular weight compounds such as
polysaccharides,proteins,pesticides,and aromatic com-
pounds(Lemmer and Kroppenstedt,1984).
Some nocardioforms can grow saprophytically on

1?l
dead cell material. The cell yields of nocardioforms were
found to be proportional to substrateconcentrationsover
a very wide range from "several" mglL to 15,000 mg/L
(Segerer,1984).Lemmer (1986) postulatedthat nocardio-
forms used these growth capabilities to produce the high
populations associatedwith activatedsludge foaming. She
proposed that the ability to grow on slowly degradable
complex organic matter provided nocardioforms a niche
that other activated sludge heterotrophs were unable to
occupy; growth on thesesubstrates(at low concentrations)
was slow and a small population of nocardioforms

H
FIGURE5.11 Relativeamountsof foamproduced on the sur-
faceofa 2-footdeepfoamtestcolumnanda l5-foot deepaeration
basinwith the samenocardioformorganismconcentrations.
Pitt and Jenkins ( I 990) developeda nocardioform filament
counting method (Table 2.3) based on the work of Vega-
Rodriguez (1983). This technique can be replicated by
observersof the samesamplewith a coefficient of variation
of about 20Vo.A similar method for M. parvicella was
developedby Mamais et al. (1988).
3. MicrobiologicalFactors
a. Factors Affecting Crowth
It is probably fair to state that more is known about how
to preventnocardioforms from growing in activatedsludge
than what causestheir growth. Lemmer (1986) presented
severalpostulatesconcerning the growth of scum-causing
resulted. When high concentrationsof readily biodegrad-
able substratebecome available, it was proposed that the
ability of nocardioforms to produce cell material propor-
tional to substrate concentration over a wide range of
substrate concentrations allowed their populations to
increase to the high levels characteristicof foaming acti-
vated sludge.
Some doubt has been cast on the validity of this
hypothesisby the pure culture chemostatdata of Blackall
et al. (1991) on the growth of two strains of G. amarae
(ASF3 from San Franciscoactivatedsludge and ASAC1
from Sacramentoactivated sludge) on acetate.Table 5.1
shows that maximum growth rates of G. amarae strains
were of the same order as those of floc-forming het-
erotrophic bacteria from a completely mixed activated
sludge plant and for the filamentous organism Tlpe 021N.
Comparedto the floc-formingZoogloea ramigera bacteria
from a selector activated sludge system, the G. amarae
strains were poor competitors both at low growth rates
(low substrate concentrations) and at growth rates close
to the respective maximum growth rates (high substrate
concentrations;Figure 5. I 2).

TABLE5.1
SteadyStateKinetic Data lor G. amaraeStrains,Type021N, and Z. ramigera
Strain p,", (d) Ko"(mg acetate/L) Koo (mg O./L) Y (g SS/gacetate)

G. amaraeSF3 2.8 0.49 0 .l 3 0.42

G. amaraeASACI 3.0 2.5 0.41
Type021N 3. 8 0.07 0.06 0.38
AS2" 2.5 0.36
Z. ramigera 5.5 0.30 0.12 0.41
u Average of floc formers from a completely mixed activated sludge system (van Niekerk et al., 1987).

Source: From Blackall, L.L. et al. (1991), Res. J. Water Pollut. Control Fecl.,63,44. With permission.
ActivatedSludgeFoamingand Control
G. amarae ASACI
012345
Substrate mgacetate/L
concentration,
FICURE5.12 Predicted relationshipof growthratesto acetate
for G. amaraestrains,Type021N,andZ. ramig'
concentration
era. (From Blackall,L.L. et al. (1991),Res.J. WaterPollut.
ControlFed.,63, 44. With permission.)
Lemmer (1986) and Soddellet al. (1998) further pos-
tulated that nocardioforms, becauseof their hydrophobic
properties and propensity to float in aeration basins, have
selectiveaccessto substrates that float, includingoils, fats,
and greases.Soddellet al. (1998) showedthat nocardioforms
can grow on vegetable oils and paraffin hydrocarbons.
Nocardioforms have often been observedadhering to and
even penetrating oil droplets (Marshall and Cruickshank,
1973; Soddell et al., 1998) and, when grown on hydro-
carbons, they can produce biosurfactants that emulsify
nonpolar substrates(Margaritis et al., 79791,MacDonald
etal., 1981; Cairns etal., 1982) and may also help in
concentrating substratesat the air-water interface.
The role of externally added surfactants in making
nonpolar substratesavailable to nocardioforms has been
discussedpreviously (Bendt et al., 1989).Further,nocar-
dioforms are adapted to dryness (Dommergues et al.,
1978) and can protect themselvesagainst solar radiation
by producing pigments (Krinsky, 1979). Taking all these
factorsinto accountsuggeststhat surfacefoam may provide
an environment where nocardioforms can compete well
in terms of substrateavailability and survival.
In support of these postulates,Matsche (1980) and
Pipes (1987) both reported that the discharge of wastes
high in oil or greaseto an activatedsludge plant can cause
a rapid increase in nocardioform populations. Pulp and
141
paper wastewatersusually have no oil and greasecompo-
nents and, in our experience, nocardioforms are rarely
observed in activated sludge systemstreating this type of
wastewater.However, on occasion, severe nocardioform
foaming has occurred due to an increasein oil and grease
from spills of hydraulic oil and from the use of oil-based
defoamers. Since there are many instances of nocardio-
form foaming in wastewaterwith low oil and greasecontent,
these substratescannot be the sole reason for excessive
nocardioform growth in activated sludge.
b. MCRT and Temperature
Anecdotal information suggests that the presence of
nocardioformsin activatedsludgeis associatedwith high
temperaturesrather than low temperaturesand with high
MCRT valuesratherthan low MCRTs. Pipes (1978) sug-
gested that nocardioform growth and foam formation in
activated sludge were problems associatedwith high
MCRT (>9 d) and high temperatures(>18'C). Wilson
et al. (1983)reportedthat successfulnocardioformcontrol
by wash-out was achievedat the 23rd Avenue plant in
Phoenix, AZ only when the MCRT was decreased to
approximately I d. Summer wastewater temperaturesat
this plant reached28oC.
Beebe (1983) reportedthat nocardioformfoaming in
the first stage of the two-stage activated sludge system at
the San Jose/SantaClara plant could be controlled by
increased sludge wasting (increasedF/M, decreased
MCRT) accompanied by chlorination of the RAS for a
short time. Typical successfuloperating parametersfor
this techniquewere increasingF/M from 0.35 to 0.5 kg
BODr/kg MLVSS/d and chlorinatingthe RAS at approx-
imately 4 kg ClrllO3kg MLVSS/d for 4 to 5 d.
Pitt and Jenkins(1990) and Cha et al. (1992) conducted
laboratory scale activatedsludge experimentsto determine
the relationship between activated sludge nocardioform
populationsand MCRT for a rangeof temperatures.In both
studies, completely mixed activated sludge units that
employed the foam trapping features illustrated in Figure
5.6a were used. Pitt and Jenkins'experimentswere done
at the Southeast Water Pollution Control Plant in San
Francisco. Cha and coworkers' experiments were con-
ducted at the SacramentoRegional WastewaterTreatment
Plant. The results of these studies are presentedin Figure
5.13 and show that nocardioform populations generally
increase with increasing MCRT over the entire range of
MCRT tested(approximately 1.5 to 20d). For the 16"C
Sacramentoreactor,nocardioform populations declined to
undetectablelevels (<104 intersections/gVSS) at an
MCRT of 2.2 d; for the 24oC Sacramentoreactor, wash-
out occurredat an MCRT of 1.5 d.
In the San Francisco reactors, the same pattern of
increasing nocardioform organism populations with
increasing MCRT and temperature was observed, but it
was not possible to obtain a wash-out of nocardioforms
142 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

)jA
EA
t>
obo
:o\
:- San Francisco
=6 25'C H
20"C H
\6
lgoc a.--a
€g l3"C o---o
8!
;e

o Sacramento
24oC a
16oC H

05101520253 0
MCRT,d

FIGURE 5.13 Comparisonof nocardioformorganismpopulationsat a rangeof MCRTs and temperaturesin bench scale-activatedsludge
units at San Franciscoand at Sacramentoin Califomia. (From Cha, D.K. etal. (1992),Water Environ. Res.,64,37. With permission.)

Influent lnfluent

Effluent

(aJ

FIGURE 5.14 Schematicdiagram of scum flows and recycle streams(a) Southeastplant, San Francisco,CA and (b) regional
wastewater treatment plant in Sacramento,CA. (From Cha, D.K. et al. (1992), Water Environ. Res., 64,37. With permission.)

to undetectable levels nor discern any difference in the
MCRT at which nocardioform populations began to
decline from their levels at very high MCRTs at tempera-
tures of 18, 20, and 25"C,. Furthermoreall the Sacramento
nocardioform counts were below the "no nocardioform
growth" level (at 13"C) at SanFrancisco.It was concluded
that the difference between these data sets was due to the
different practices employed at the two plants for dealing
with removed foam and the recycle streamsfrom the solids
handling processes(Figure 5.14).
At the time of the study in San Francisco, the foam
from the mixed liquor channelswas returned to the head-
works. At Sacramento,secondary scum from the mixed
liquor channel was first sent to the WAS-dissolvedair flo-
tation thickenersand thenceto the anaerobicdigesters.After
anaerobic digestion, all the digester contents were trans-
ferred to solids retention basins that had sludge residence
times on the order of 5 y. Supematant from the solids
retention basinsand subnatantfrom the dissolvedair flota-
tion units were both returned to the headworks.These dif-
ferencesin foam handling and recycle streamssuggestthat
seeding of the influent with nocardioforms from removed
and recycled foam and from solids treatmentprocessrecy-
cle streamswas the causeof the higher nocardioform pop-
ulations and the inability to wash out nocardioformsin the
San Francisco activated sludge. Attempts to confirm this
by detectingnocardioform filaments in the primary effluent
entering the San Francisco activated sludge system were
not successful(Pitt and Jenkins, 1990).
The MCRT values obtained for nocardioform washout
in thesestudies agreedvery well with those that employed
at the Sacramento and San Francisco plants to control
foaming. Table 5.2 presents these data, and Figure 5.15
shows an Arrhenius equation plot for a combination of the
Sacramentopilot plant data and the full-scale plant data.
These data can be utilized to determine the MCRT
ActivatedSludgeFoamingand Control 143

TABTE5.2
Wash-outMCRT (MCRTUsed for NocardioformOrganism
Control) at ActivatedSludgePlants
Source of Data Temperature(oC) MCRT (d)
Sacramento,CA - Pilot Plant 16 2.2
Sacramento,CA - Pilot Plant 24 1.6
Sacramento,CA - Full Scale (Summer) 22 1.8
Sacramento,CA - Full Scale (Winter) 18 2.2
San Francisco, CA - Full Scale 20 1.1
u Maximum month flow

Source: From Cha, D.K. er al. (1992), Water Environ. Res., 64, 37. With permission.

Temperature, oC
16.8 12.7
1.0

iE -0.2
,.{
e
) -nd 1.5 e

I -
O
E -n6 1.8>
x

! -r.s 2.2

- 1. 0
3.35 3.40 3.45 3.50
l/temperaturex 103

FIGURE5.15 Relationship of nocardioform (From Cha,

organismmaximumnet growth rate in activatedsludgeto temperature.
D.K. et al. (1992),WaterEnviron.Res.,64,37.With permission.)

required to wash out nocardioforms from activatedsludge
over a range of temperatures.
Soddelland Seviour(1995)suggestthat cautionshould
be used when employing such an Arrhenius plot to predict
the growth rate (MCRT) at which nocardioformswill wash
out from activated sludge becausethey showed that some
nocardioforms isolated from activatedsludge grew at tem-
peraturesof 4oC and others grew at temperaturesbetween
40 and 50'C. They argued that changing the temperature
of an activated sludge to eliminate nocardioforms may
actually result in the replacementof one nocardioform by
another and a continuation of the foaming problem. They
indicated that Rhodococcus spp. grew better than other
nocardioforms at low MCRTs and low temperatures.
Despite this argument, it has been our experiencethat the
use of low MCRT for eliminating nocardioforms from
activatedsludgehas been uniformly successfulin practice.
c. pH
Cha et al. (1992) examined the effect of pH on nocardio-
form populations in activated sludge using completely
mixed laboratory activated sludge units operated over a
rangeof pH valuesbetween6.0 and 7.5 and at MCRT values
of 3 and 8 d. Figure 5.16 shows that at both MCRTs, pH
influencedthe nocardioform counts in the activatedsludge.
This influence was greater at the 8-d MCRT and indicated
an optimum in the region of pH 6.5; a similar trend was
evident in the data at an MCRT of 3 d. The value of the pH
optimum is significant from the viewpoint of nocardioform
occurrencein full-scale activatedsludge systems.
In their survey of nocardioform foaming in prototype
activatedsludge plants, Pitt and Jenkins (1990) found that
every one of the six oxygen-activated sludge plants
responding to their questionnaire experienced nocardio-
form foaming. The averagemixed liquor pH value of the
air-activatedsludge plants was 7; for the oxygen-activated
sludge plants the averagepH value was 6.5. Lower mixed
liquor pH values are typically encounteredin oxygen-acti-
vated sludge plants becausethe mixed liquor is in contact
with a gas phase high in COr. In addition, it is important
to note that the aerationbasins in oxygen-activatedsludge
systems always have subsurface withdrawal of mixed
1 44 M anua lo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
8
a
v) l
.J>
=-< 6
ix s T.
Fa
I
-f
xo
z+
0. 1
5. 5 6.0 6.5
FIGURE 5.16 Effect of pH on nocardioform organism populationsin bench scale-activatedsludge units. (From Cha, D.K. etal.
(1992), WaterEnviron. Res.,64,37. With permission.)
liquor so that foam trapping occurs. The relative impor-
tance of foam trapping and mixed liquor pH on the high
occurrenceof nocardioform foaming in oxygen-activated
sludge is not known precisely.However,an examination
of Figure 5.7 and Figure 5.16 suggeststhat foam trapping
might well be the more important factor. Figure 5.7 sug-
gests that the installation of trapping features in the labo-
ratory pilot plant increasedthe nocardioform count approx-
imately ten-fold.Figure 5.16 suggeststhat a changein pH
from about7.0 (typical pH of air activatedsludgesystems)
to 6.5 (typical pH of oxygen activatedsludge systems)
would resultin an increasein nocardioformcount ofabout
2O7oor about 1/50 of the effect on nocardioform count due
to foam trapping.The occurrenceofboth low pH and foam
trapping in the same system should provide excellent con-
ditions for nocardioform growth and retention.
It is often observed that nocardioform foaming inci-
dents often coincide with the onset of nitrification. This
observation can be explained in terms of both MCRT and
pH effects.First, the onset of nitrification indicatesthat
the aerobicMCRT is sufficientlyhigh to allow the growth
of nitrifying bacteria.Second,the consumptionof alka-
linity by nitrification can result in depressedpH which
also favors the growth of nocardioforms.
III. NOCARDIOFORM CONTROT
A. NocnnororoRM
GRowrHrNAcnvATED
Sruocr
1. lntroduction
The abilities of the varioustypes of selectors(aerobic,
anoxic,andanaerobic)to controlnocardioformgrowthin
activatedsludgehasbeenexaminedby a combinationof
pure culture experimentson G. amarae and activated
sludgeexperiments at laboratory,pilot andfull-scale.
Pr oblem s
M C R T =8 d o . . . o
MCRT=3d o--o
.I
?
'1-0 7.5 8.0
pH
Blackall et al. (1991) grew G. amarae ASF3 and
ASACI in a chemostatwith acetateas a solecarbonsource
over a wide range of dilution rates. The organisms were
removedfrom the chemostatand their batch acetateuptake
rates determinedunder aerobic conditions (DO present),
anoxic conditions (no DO present;NO;-N or NO|-N
present) and anaerobicconditions (no DO, NOf-N, or
NOt-N present).Batch substrateuptake rates suggest
how well an organismwill fare in a selector;high substrate
uptake rate provides a competitive advantage.
2. Aerobic Selectors
In Figure 5.17, the aerobicbatch acetateuptakerates for
G. amarae are comparedto thoseof Zoogloea ramigera -
a floc former found in selectoractivatedsludge and shown
by van Niekerk et al. (1987)to be responsiblefor the rapid
uptake of soluble COD in an aerobic selector.The varia-
tion of batch acetate uptake rate with organism growth
rate (dilution rate) is quite different for these two organ-
isms. Of significance for nocardioform organism control
is the observation that only at very low dilution rates is
the G. amarae ASF 3 unbalanced growth acetateuptake
rate greater than that of Z. ramigera.
It should be recognized that the organism dilution
rates usedin thesestudies(range I to 5/d) are significantly
higher than those used in most full-scale activated sludge
systems.Also, the uniform growth environment provided
in a chemostat differs from the temporally and spatially
variable environment provided in a full-scale system.This
latter factor may result in differences in the physiological
stateof the organisms.Nevertheless,the data presentedin
Figure 5.17 suggestthat an aerobicselectormay be more
effective in controlling nocardioforms in activated sludge
at a moderate MCRT than at a hish MCRT.
ActivatedSludgeFoamingand Control 145

ondary clarif,er weir. The aerobic selector was judged to

be functioning properly on the basis of its rapid soluble
COD uptake rate in a batch test, control of SVI, and
observation of typical amorphous zoogloeal colonies in
the flocs.
1J Figure 5.18 shows that at a 5 d MCRT, the nocardio-
d form count in the completely mixed control unit increased
from an initial value of 106intersections/gVSS to 5 x 106
. ! : lo
EU)
dC)
after about 2 or 3 MCRTs and thereafterremainedbetween
o
--:
3.5 x 106and 6 x 106until the end of the experiment (after
E
o:
Eg 7.5 MCRTs). The nocardioform count in the aerobic selec-
9Y
dx
tor unit, on the other hand, declined gradually from its
!2 =,
EE6 initial value of 106 to a value of 105 at the end of the
experiment. The results indicate that, at a 5-d MCRT, the
aerobic selector controlled nocardioforms.When the aer-
L Z. ramigera obic selectorwas operatedat a 10-dMCRT (Figure 5.19),
O G. amarae ASF3 effective nocardioform control was not observed.After one
o G. amarae ASACI MCRT, nocardioform counts in the CSTR control system
fluctuatedbetween 107and 1.7 x lO7.while those in the
01234 5 aerobic selector unit were somewhat lower (in the range
Dilution
rate,d-l of 5 x 106to 107intersections/gVSS) over the 9-MCRT
experiment.While the aerobic selectornocardioform pop-
FIGURE5.17 Comparison of G. amaraeandZ. ramigeratran- ulations were in a somewhatlower range than those in the
sient stateacetateuptakeratesover a rangeof dilution rates. control system, Figure 5.19 shows that toward the end of
(FromBlackall,L.L. et al. (1991),Res.J. Pollut.ControlFed.,
the experiment (from 7 to 9 MCRTs), the nocardioform
63, 44. With permission.)
counts in both units were very close. On this basis, it can
be stated that effective nocardioform control was not
Cha et al. (1992) investigatedthe ability of an aerobic achievedby an aerobic selector at an MCRT of l0 d.
selector to control nocardioform populations in activated Operating results from the 27 -MGD Upper Occoquan
sludge at MCRTs of 5 and l0 d using a four-compartrnent Sewage Authority (UOSA) Water Reclamation Plant
selectorat a temperatureof 20'C. A DO concentrationof at (WRP) in Centreville, VA are consistent with the results
least5.0 mg/L was maintainedin all selectorcompartments of Cha etaL (1992)'.The UOSA WRP utilizes aerobic
and in the main aerationbasin.All units had nocardioform selectorsto control filamentousbulking (Daigger et al.,
foam trapping features, i.e., subsurfaceaeration basin 1985;Daigger and Nicholson, 1990).The effectivenessof
mixed liquor withdrawal and a baffle in front of the sec- the selector in controlling the growth of filamentous

(t MCRT=5d CSTRcontrol o----o

U)
;) Aerobic selector o------o
EE
ov

frx

oo

xa

0.01
0123456789
No. ofMCRTs

F|GURE 5.1 8 Effect of aerobic selector on nocardioform organism population at a 5-day MCRT. (From Cha, D.K. et al. (1992),
Water Environ. Res., 64,37. With permission.)
146 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

MCRT = 10d CSTRcontrol o------r

U)
U) Aerobic selector o-----o
.r>
=-<
9v

:Ya
s .9
l0
EE

Hb
z*-

0.01
0123456
No. ofMCRTs

FIGURE 5.19 Effect of aerobic selector on nocardioform organism population at a l0-day MCRT. (From Cha, D.K. etal. (1992),
WaterEnviron. Res.,64,37. With permission.)

organisms is demonstrated by SVIs routinely below
100 mllg in the full-scale system.Operatingexperience
indicates that the selectors are effective in controlling
nocardioform foaming in the summer (wastewater tem-
perature generally above 20"C) when the MCRT is main-
tained within the 5- to 8-d range. However, nocardioform
foaming occurs when the MCRT exceeds l0 d, even
though the selector is operated. Nocardioform foaming
does not generally occur in the winter (wastewater tem-
perature l5oC or lower), even though MCRTs of 10 to
15 d are maintained.
3. Anoxic Selectors
At MCRT values >10 d, activated sludge systemsoperat-
ing at temperatures of >20oC usually will nitrify. This
offers the opportunity to use an anoxic selectorfor nocar-
dioform control. This possibility was investigatedby Cha
etal. (1992) at an MCRT of 12 d. The anoxic selector
design described earlier was used. It was shown to func-
tion satisfactorily as a selector by soluble COD uptake
and NOr-N removal over the selector of approximately
40 mglL and approximately 5 mg/L, respectively,control
ofSVIto <100 mllg, and presenceofamorphous zoogloeal
colonies in the activated sludge.
Figure 5.20 shows that the CSTR control unit exhib-
ited nocardioform counts that ranged from 106to 4 x 106
intersections/gVSS over a period of 7 MCRTs (generally
increasingthroughout the experiment).In the sameperiod,
the anoxic selector system nocardioform count remained
between 5 x lOs and 106intersections/gVSS. After 66 d
(5.5 MCRTs), the contents of the reactors were inter-
changedso that the CSTR control aerobic activatedsludge

MCRT= 12d q
b0
a
a CSTRcontrol H .E
q.l
-r) Anoxic selectorG--o
E{9 !l

9v

FX

EZ

Hb

0.01
012345678
No. ofMCRTs

FIGURE 5.20 Effect of anoxic selector on nocardioform organism populations. (From Cha, D.K. et al. (1992),Water Environ. Res.,
64, 37. With permission.)
ActivatedSludgeFoamingand Control
t20
tr
o (0.74 m3/s)Rock Creek,OR plant usesan anoxic selector
----o-__?__q
trn
6Aa
E
.E
UU
AA
ob
'E OU
'a
Aerobic rate
;,4 0
A Anoxic (NO2)
o Anoxic (NOj)
tr Anaerobic
05101 5 2 0
Time,h
FICURE5.21 Transientstateacetateuptakeby G. amarae
ASF3 underaerobic,anoxic,and anaerobic conditions.(From
Blackalf, L.L. et al. (1991),Res.J. WaterPollut.ControlFed.,
63, 44. With permission.)
was placed in the anoxic selectorsystem and vice versa.
The nocardioformcount in the activatedsludgefrom the
control unit immediately and rapidly decreasedunder
anoxic selector conditions; the anoxic selector sludge
placedin the CSTR control unit showeda gradualincrease
in nocardioform count to levels above those observed
while this sludge was in the anoxic selectorunit. These
experiments suggestthat properly designed anoxic selec-
tors should be effective in controlling nocardioforms in
activatedsludge.They are also consistentwith the pure
culture data of Blackall et al. (1991) on G. amarae who
showedthat G. amarae could not take up acetatein anoxic
batch tests(Figure 5.21) and that it denitrifled at rates two
to three orders of magnitude lower than for Z. ramigera
(van Niekerk et al., 1987) and only incompletely to
NO2-N, rather than N, gas (Table 4.8).
Sezgin and Karr (1986) convertedthe first passof a
four-pass aeration basin in a nitrifying activated sludge
plant at Utoy Creek, GA to an anoxic zone by reducing
the air flow to the coarsebubble diffusers. On three occa-
sions out of five, Richardset al. (1989) reportedthat this
was successfulin reducing foam coverageof the aeration
basins.A truly anoxiczonewas not obtained(0.2 to 0.5 mg
DO/L was present)becauseof the use of aerationfor mix-
ing; also low aeration rates in the anoxic zone limited the
extent of nitrification in the plant, meaning that less NO,-
N was recycled back to the anoxic zone in the RAS.
147
Results from other full-scale plants utilizing anoxic
selectors indicate the dual impact of an anoxic selector
and of foam trapping and recycling. The 17-MGD
for filament control and alkalinity recovery (Daigger and
Nicholson, 1990).An accumulationof nocardioformfoam
is observedduring periodswhen secondaryscum is recy-
cled to the plant headworks. Similarly, nocardioform
foaming was observed during pilot studies for the
40-MGD (1.75 m3/s)VIP plant in Norfolk, VA (Daigger
et al., 1988)and in the completedfull-scalefacility. Both
anoxic and anaerobicselectorswere incorporated into the
VIP pilot and full-scalefacilities.Foam trappingoccuffed
in both the pilot and full-scale units and may explain the
nocardioform foaming incidents. Nocardioform foaming
at other full-scale nutrient removal plants that incorporate
anoxic zones for nitrogen removal is often attributed to
foam trapping causedby the placementof baffles.Taken
together,theseresults indicate that anoxic selectorsmay
help to control nocardioform growth but their positive
effects may be offset by foam trapping and recycling.
4. Anaerobic Selectors
Data on the effectivenessof anaerobicselectorsin con-
trolling nocardioformgrowth in activatedsludge are not
asextensiveas datafor aerobicand anoxic selectors.How-
ever, as a result of the work of severalinvestigators(Pitt
and Jenkins,1990;Blackall etal., 1991;Mamais, 1992;
and Fainsodet al., 1999), it is possibleto conclude that
anaerobic selectors intended to control nocardioform
growth in activatedsludge must be operatedto allow the
establishment of EBPR.
This can be judged using several criteria including
efficient phosphorusremoval, initial anaerobicreleaseof
dissolvedorthophosphate from activatedsludgeaccompa-
nied by dissolved COD uptake, presence of Neisser-
positivecell clumps in activatedsludgeflocs, and elevated
activatedsludge total P content (>27o PIYSS). The con-
clusionis supportedby the following experimentalresults.
Blackall et al. (1991) showedthat G. amarae neither
took up acetate(Figure 5.21) nor grew under anaerobic
conditions, indicating that anaerobic selectors should
provide conditions where nocardioforms can be out-
competed by microorganismsthat can accesssubstrate
anaerobically.
Laboratory-scaleexperiments(Pitt and Jenkins,1990)
at the San Francisco South East Water Pollution Control
Plant failed to demonstrateconsistentcontrol of nocardio-
form organisms by an anaerobic selector. Their experi-
ments were confoundedby the growth of Thiothrix spp. in
the test system activated sludge. Pagilla (1994) overcame
thesedifficulties,using the findings of Gabb et al. (1989)
to eliminate filamentous organism seeding by S. natans,
Type 1701andThiothrix spp.from feed transmissionlines
148 M anu a lo n C a u s e sa n d C o n tro lo f Ac t i vatedS IudgeB ul ki ng,Foami ng,
and storagevessels.Pagilla's experimentsutilized 40-L
reactors consisting of a control reactor with four aerated
l0-L compartments in series and an anaerobic selector
reactor with a four-compartment selector (2.5 L, each
compartment) followed by three aerated 10-L compart-
ments in series.Anaerobic selectortotal HRT was 50 min.
Anaerobic selectorcontentswere mixed and kept free
of DO by purging with zero gradeNr gas in all four stages.
DO was 8 mg/L in the first aeration basin stageand about
4.0 mglL in the final stage.The aerationbasinsand sec-
ondary clarifiers had the following foam trapping features:
subsurfacemixed liquor transfer between aeration basin
and secondary clarifiers; scum baffle in secondary clari-
fier; and secondary clarifler scum returned to aeration
basin. Once bulking-free operationhad been established
in the control and EBPR had beenestablishedin the anaer-
obic selector, nocardioform filament counts remained
below the level of detectionof lOaintersections/gVSS in
the anaerobic selector system. In the control, they
increasedfrom below detection levels to the 105to 106
intersections/gVSS range. These experiments demon-
stratedthat an anaerobicselectorexhibiting EBPR (and
operatedunder conditions of MCRT = 2.4 d, temperature=
20"C, pH = 6.3, anaerobicselectorHRT = 50 min, and
total reactorHRT = 3.3 h) can control nocardioformorgan-
ism growth in an activatedsludge plant that has foam-
trapping features.
Full-scale anaerobicselectorexperimentswere con-
ducted at the San FranciscoSouth East Water Pollution
Control Plant (SEWPCP) from May through October
1987,May throughOctober1988,1992,and1994through
1995 (Pitt and Jenkins, 1990; Mamais, 1992; Fainsod
et al., 1999).The SEWPCPmixed liquor pH and temper-
ature during thesestudieswere in the rangesof 6.3 to 6.8
and l7 to 19"C, respectively).This oxygen-activated
sludgeplant includessix equal-size-compartment aeration
basinsand eight treatmenttrains. For the experiments in
1987 and 1988. four trains were convertedto the A/O
process by making the first compartment anaerobic
(replacing the 50-Hp aerator with a 10-Hp submersible
mixer and feeding the Or gas to the secondcompartment).
The sizes of the interstageopenings were decreasedto
preventback-mixing betweenstages.
During 1987, the three following anaerobicselector
operatingconditionswere examined:MCRTs of 1.4,2.0,
and 2.3 d; initial selectorF/Ms of 6.6,'7.8,and 13.2kg
BOD./kg MLVSS/d, and average hydraulic detention
times of 13, 19, and 25 min. While it was possible to
separatethe plant into two halves, the results were con-
founded by cross-contaminationbetween the control and
the anaerobic selector systemsby spilling of control RAS
over a common wall into anaerobic selector RAS and
spilling of fbam from the control mixed liquor into the
anaerobicselectormixed liquor over a dividing gate in the
Pr oblem s
and Other S ol i dsS eparati on
mixed liquor channel.The RAS cross-contamination was
rectifiedfor the 1988testing.
Nocardioform organism counts, secondary clarifier
foam coverage,and foam height in a foam test were some-
what lower in the anaerobic selector system than in the
control first stage.A small (0.8 to 2.4 mg P/L) soluble
ortho-P releaseoccurred in the anaerobic selector system
but not in the control. At no time did the control or anaer-
obic selector systems show any differences in overall
phosphateremoval.
The full-scale anaerobicselectortest was repeatedin
summer 1988 using the following anaerobicselectorcon-
ditions: initial F/Ms of 1.5, 1.6, and 2.1 kg BOD./kg
MLVSS/d, hydraulic detentiontimes of 9, 10, and 13 min,
and overallMCRTs of 1.6,2.1. and 1.5 d. Cross-contam-
ination due to foam spilling from the control systeminto
the anaerobicselectorsystemoccurredfor the first 5 wk
of the experiment.
The 1988 results again showed that the anaerobic
selectorhad slightly lower nocardioformorganismcounts,
secondaryclarifier foam coverage,and foam height in a
foaming testthan in the control system.Anaerobicsoluble
ortho-Preleasewasonly 2.1 to2.9 mgPlL in theanaerobic
selector system and the two systems again showed no
differencesin overall phosphateremoval efficiency.
Both the aboveanaerobicselectorexperimentsof Pitt
and Jenkins (1990) were carried out at MCRT values in
the range 1.4 to 2.3 d - operatingvaluesthat had been
employedby plant staff to provide some degreeof foam-
ing control by nocardioformorganismwash-out.
EBPR is usually accompaniedby the uptake of bio-
degradablesoluble organicsin an anaerobicselectorand
without this uptake of soluble organics (and therefore
without EBPR), the beneficialeffectsof the selectorwill
not be exhibited.Mamais and Jenkins(1992) determined
the minimum MCRT at which activatedsludgewill exhibit
enhancedbiological phosphorusremoval (EBPR) over a
range of temperatures(Figure 5.22). These data can be
usedto establishthe minimum MCRT at which an anaer-
obic selectorrelying on EBPR can be operatedfor a range
of temperatures.Figure 5.23 shows that the wash-out
MCRT for nocardioforms in activated sludge (Cha et al.,
1992) is very close to the wash-out MCRT for EBPR at
a given temperature.Thus, a plant operatedat a low MCRT
for nocardioform organism control (such as the San Fran-
cisco plant during Pitt and Jenkins' 1990 experiments)
will very likely be unable to consistentlysupportEBPR
and therefore would not provide satisfactory conditions
for an anaerobic selector.
To test this hypothesis Mamais (1992) operated the
SanFranciscoplant at MCRT valuesranging from approx-
imately 1.5 to 2.5 d and with anaerobicselectorHRTs in
the range of 15 to 25 min. By the time of these experi-
ments,the entire facility had beenconvertedto an anaerobic
selectorplant so that a side-by-sidecontrol and anaerobic
 ActivatedSludgeFoamingand Control 149

Temperature, oC
25.5 21 t'7
0.0 1.0

ln (netp.u*) = 4366(IIT) + 14.2 t.2 E

b Slope= E"rR-+ Eu= 36.3L17.o1" I
o
-0.4 1.5u

> -0.6 l .d E

3
-0.8

- 1. 0

-1.2
33.5 34.0 34.5 35.0 35.5
x 104,l/oK
l/Temperature

FIGURE 5.22 Effect of temperatureon the washoutMCRT for enhancedbiological phosphateremoval in activatedsludge.(From
Mamais, D. and Jenkins,D. (1992), Water Sci. Technol.,26:5-6,955. With permission.)

the range from 2.1 to 3.2 d (average,2.5 d). Period 2 had

MCRTs in the range from 2.6 to 3.7 d (average,3.2 d).
l0 r-r Period 3 had MCRTs in the rangefrom 2.8 to 4.6 d (average
Nocardioformorganismwashout
3.5 d). Periods 1 and 2 had influent flows averaging 61
6
€ MGD and rangingfrom 50 to 96 MGD (average,2.7 m3ls:
EBPR and nitrification range,2.2 to 4.2 m3ls)typical of the dry season.Period 3
-,
Q
influerlt flows were strongly influenced by rainy weather;
A OnlyEBPR;
the averagewas 93 MGD and rangedfrom 54 to 211 MGD
no nitrification
(average,4.1m3/s;range,2.4to 9.2 m3/s).Secondaryefflu-

L- No EBPR:-=:
no nitritication
lo t2 t4 16
,:t
";1,.".1'.
FIGURE5.23 Limiting MCRT valuesfor nitrification,EBPR,
and nocardioformgrowthover a rangeof temperatures.
selector comparison was not possible. As the overall
MCRT increasedfrom 1.5 to 2.5 d, EBPR becameestab-
lished with anaerobic ortho-P release increasing from
approximately 0 to 10 mgPlL and effluent P concentration
decreasingfrom approximately 4 to 2 mg P/L. Nocardio-
form organism counts averaged approximately 3 x 106
intersections/gVSS) which is significantlylower than val-
ues obtainedby Pitt and Jenkins(1990).
ln 1994 and 1995, Fainsod etal. (1971) conducteda
fourth series of full-scale anaerobic selector experiments
on nocardioforrn organism control at the San Francisco
plant. The MCRT of the entire anaerobic selector plant
was gradually raised from 2.3 d to 3.5 d.
Figure 5.24 shows a series of plots of all 1994 and
1995 datapoints for influent flow, MCRT, soluble ortho-P
(primary effluent, secondaryeffluent, and anaerobicselec-
tor release), and mixed liquor nocardioform filament
counts. The experimental data can be divided into three
periods on the basis of MCRTs. Period t had MCRTs in
ent temperatureaveraged20'C (range, 17.5 to 21.7.'C) and
with no trends observedthroughout the experiment.
24 26 28 30 EBPR occurredto some degreethroughoutthe entire
experiment.The effluent soluble ortho-P concentration
decreasedfrom PeriodsI and2 (averages,1.3 and l.l mg
P/L, respectively)to Period 3 (average,0.4 mg P/L), indi-
cating that EBPR becamebetterestablishedas the MCRT
increased.The anaerobicselectorsoluble ortho-P release
did not show the sametrend and may have been influenced
by changesin influent solubleortho-Pconcentrationsaris-
ing from influent dilution by storm water (especially in
Period 3).
During Periods I and 2, the activated sludge total P
content gradually increasedfrom 1.3 to 4Voby weight on a
VSS basis.This provided further evidenceof the establish-
ment of EBPR as the MCRT was raised from 2.5 to 3.2 d.
The lowest nocardioform filament count (0.9 x 106inter-
sections/gVSS) and the lowest averagesecondaryeffluent
soluble ortho-P concentration(0.4 mg P/L) were both asso-
ciated with the highest averageMCRT tested(4.6 d).
The mixed liquor nocardioform filament counts
decreasedfrom their values in Periods I and 2 (Period I
average,2.9 x 106intersections/g VSS; range, 1.3 to 5.2x
106 intersections 1 g VSS; Period 2 average,2.6 x 106
intersections/gVSS; range, 1.8 to 3.5 x 106intersections/g
VSS) to thosein Period 3 (average,1.4x 106intersections/g
1 50 Pr oblem s
M anua lo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on

F
U lA.
z ^AA
Period 1
Period
2 Period 3

i 200
o

Ej too il,
0

Primary effluent
20 I st stagerelease
o
Secondaryeffluent
6<
l0
I,NV!; :\- r -
j _ "/\
r i ,.* ..-
----' - ,- - - - 'l -
.A .lL " ,
--i - ,- ,:" -_ a .* ._ -s -
0 ^

5
Ea 4
6tt
U>
3
2
fr.= I

z= Jun Aug Apr Jun Aug Oct Dec

+ 1994 tgg5 1996

Time.Months

FIGURE 5.24 Resultsof full scale anaerobicselectorexperimentsconductedat San Franciscoin 1994 and 1995. (From Fainsod,
A. et al. ( 1999),Water Environ. Res.,71, 1I 5. With permission.)

VSS; range,0.9 to 2.8 x106intersections/g VSS). Increas-
ing the overallMCRT from 2.5 to 3 .5 d in the SanFrancisco
anaerobic selector plant greatly reduced nocardioform
organismlevels and nocardioform-associatedproblemsbut
it was never possible to reducethe nocardioform organism
levels to nondetectableas was achieved in the laboratory
experimentsof Pagilla (Fainsod etal., 1999).
Figure 5.25 presentsthe entire set of datafrom all the
full-scale experiments at San Francisco. The average
MCRTs and operating temperaturesof all the nocardio-
form organism control experiments (laboratory and full-
scale) are plotted in Figure 5.26 together with (1) their
successat reducing nocardioform counts to below detect-
able levels and (2) the ranges at which EBPR and nitrifi-
cation occur in activated sludge.
The open symbolsin Figure 5.26 representexperiments
in which nocardioformscould still be detectedin the acti-
vated sludge.Solid symbolsrepresentexperimentsin which
nocardioform organismswere reducedto below detectable
levels (nocardioform filament counts <104 intersections/g
VSS). At the San Franciscoplant, at a temperatureof 20'C,
the full-scale experimentsspannedthe entire MCRT range
for EBPR without nitrification and nocardioform f,lament
counts could not be reduced to below detectablelevels.
However,at the sametemperature(20"C) and at an MCRT
of 2.4 d, nocardioform organismswere reduced to below
detectablelevels in the Sacramentopilot plant system.
Possibledifferencesbetweenthe pilot-scaleoperating
conditionsat the Sacramentoplant and thoseat San Fran-
cisco plant that might accountfor theseresultsinclude the
following pilot-scaleexperimentalconditions:
. Absence of nocardioform-organism containing
return streamsin the influent
. Higher anaerobic selector HRT
. More effective compartmentalization in the
anaerobic selector
All these factors may be significant. The influence of
nocardioform recycles has already been discussed.The
anaerobic selectorsemployed in the San Francisco plant
experiments consisted of only one compartment with an
HRT of only 17 to 24 min. In the Sacramentopilot plant,
the anaerobic selectorscontained four equally-sized com-
partments in series with a total HRT of 50 min.
ActivatedSludgeFoamingand Control 151

6
5 Average for period

l3
(J
j+
-rt
1
0

$ zoo
i 150
E 100
o

E so

Primary effluent

S 20 I st stagerelease
bo Secondary effluent

o lu

V) l''i,'r
0
7 T .-...
2(t
aa
5>
6
5
I
ll
I
Average for period

/
5s
xv
z
3
2
I
0
l\u.*-
Apr Dec Aug Apr Dec Aug Apr Nov Jul Mar Nov Jul Mar Nov

- 87 -{- 88 --.r- 89+ 90 + 9t + 92 + 93 + 94 -F95 + 96-

Month andYear

FIGURE 5.25 Resultsof all full scaleanaerobicselectorexperimentsat San Franciscofrom 1987 through 1995.(From Fainsod,A
et al. (1999),WaterEnviron.Res.,7l,ll5. With permission.)

These results suggest that a properly designed and
sized anaerobic selector activated sludge system exhibit-
ing EBPR in a systemwithout nocardioform recycling can
control nocardioform organisms in an activated sludge
system with foam-trapping features. The association of
EBPR effectivenesswith nocardioform control ability is
illustrated in Figure 5.2'7; as effluent soluble ortho-P
decreases,so do the activatedsludge nocardioform counts.
5. Classifying Selectors and Selective
Foam Wasting
Reference was made earlier to the analogy that can be
drawn between nocardioform foaming in activated sludge
and the use of hydrophobic collectors for the separation
of minerals from ores by selectiveflotation. This principle
has been used to remove nocardioform organisms from
activated sludge. Pretorious (1987) suggestedthat the
buoyancy of nocardioforms could be used to selectively
float them from settleableactivatedsludge flocs in a "clas-
sifying selector."He suggestedthat if the float from such
a selector were not returned to the aeration basin, nocar-
dioforms would be eliminated.
Pretoriousand Laubscher(1987) investigatedthis prin-
ciple in 200-L mechanicallyaeratedactivatedsludgeplants,
one of which was equipped with a 16.7-L fine bubble flo-
tation cell between the aeration basin and the secondary
clarifier. Foaming in the classifying selector system was
virtually eliminated after about 2 d while the control system
continued to foam severely.While the flotation cell had no
152 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

- O - Nocardiofom orgmism washout

Region III (cha, et al., 1992)
(EBPR md nitrification)
I Pagilla,1994
D Mmais, 1992
8 A Pitt md Jenkins, 1987
O Pitt and Jenkins, 1988
O Period l,1994-1995
F
O Period 2, 1994-1995
Q
V Period 3. 1994-1995

Region II
(EBPR; no nitrification)

ur

Region I
(no EBPR; no nitrification)

t0 t2 t4 16 18 20 22 24 26 28 30
Temperature,
"C

FICURE 5.26 MCRT and temperature regions for EBPR, nitrification and nocardiofonn growth in anaerobic selector activated
sludge. Solid symbols represent experiments in which nocardioforms were not detected. Open symbols represent experiments in
which nocardioformswere detected.(From Fainsod,A. et al. (1999), Water Environ. Res.,'71,115. With permission.)

U)
(/)
I

,i
a

+g

p
d
o Effluent soluble orthoP
o Nocardioform counts
z
-+0
1. 5 2.0 4.0
MCRT.d

FIGURE 5.27 Variation of average nocardioform counts and effluent soluble orthoP concentrations with MCRT for full scale
anaerobicselectorexperimentsat San Francisco.(From Fainsod,A. et al. (1999), WaterEnviron. Res.,7l,l15. With permission.)

long-term effect on the MLSS level, a decrease(from about
4500 to 4000 mg/L) was noted during the first few days
after the installation of the flotation cell. This observation
suggeststhat a flotation cell applied to a long-standing
severenocardioform foaming problem with much accumu-
lated foam may initially waste out more than the desirable
amount of solids inventory. In such cftcumstances,it may
well be good practice to vacuum off as much accumulated
foam as possible,then start using the flotation cell.
Selectivefoam wasting was employed successfullyby
Richards et al. (1990) at the Utoy Creek, GA activated
sludge plant. This plant was operated in a sludge reaera-
tion/step-feed mode (Figure 5.28) with RAS only in the
first pass of the aeration basin. Foam was allowed to pass
from the aeration basin into an unusedbasin where it was
concentratedby collapsing. Foam SS concentration was
2 to 3Voand dewateredwell using the centrifugesnormally
used for WAS. Using selective foam wasting to control
foam allowed the plant to increaseaerationrate and MCRT
values (from 6 to 2O d) so that complete nitrification was
possible for the first time. Once the MCRT was increased
to >20 d, the severe foaming ceased and foam wasting
was not necessary.From a previous article about this plant
(Sezgin and Karr, 1986), it is suspectedthat an industrial
waste containing a slowly degradable surfactant was
present which was only degraded completely at high
ActivatedSludgeFoamingand Control 153

7 lection sump). If pumping is required to empty the col-
Fou
Influent
FIGURE5.28 Planviewof aeration basinswith selectivefoam
wasting,UtoyCreek,GA. (FromRichards, T. et al. (1990),Res.
J. WaterPollut.ControlFed.,62,914.With permission.)
MCRT. The nocardioform foaming may have been stabi-
lized by the presenceof the surfactantat the lower MCRT.
Parker et al. (200 I , 2003) reported the use of selective
foam wasting (classifying selectors) at seven activated
sludgeplants(Table5.3). They emphasizethe importance
of removinga small amountof liquid almostcontinuously
from the top 1 to 3 cm of a mixed liquor or RAS stream.
This layer can be removed through devices such as down-
ward opening gates or side weirs on channels. Such
devices are best located where the natural flow tends to
direct foam, where the liquid level does not vary greatly,
and where surface turbulence is not excessive.
One or more removal locations may be required and
foam trapping at locations other than where it is removed
should be avoided. Surface wasted material should be
combined with WAS for treatment and the solids con-
tained in it should be accounted for in sludge wasting
calculations. Any collection sump for removed surface
material should be designed to allow for its complete
removal when emptied. Failure to do this will transfer a
foaming problem from one location to another (the col-
lection sump, the pump should be able to break suction
rather than to allow vortex development that would pre-
vent floating material removal. The pump should be able
to pass liquid containing entrained air. Suitable pumps
include submersiblesolids handling, positive displace-
ment diaphragm, self-priming, and air-lift types. Pump
capacitymustbe high enoughto pump down the collection
sump, break suction, and remove floating material at all
but peak flows. Pump control devices must be able to
function in liquids containing entrained air.
Parkeret al. (2001, 2003) showedthat the installation
of a classifying selector on the RAS channel of the
29-MGD (1.3 m3/d) Haskell R. Street plant in El Paso,
TX virtually eliminated activated sludge foaming and
reduced nocardioform organism counts from I to 2 x 106
intersections/eVSS to 6 xl04 intersections/sVSS.

TABTE5.3
ActivatedSludgePlantswith SurfaceWastingin Operation
MMF" Capacity, Aeration Basin Surface Waste
Plant Location MGD (m3/d) or Channel Type Surface Wasting Location Disposal Location

Sacramento,CA 180( 8. 1) Oxygen/coarse bubble MLb distribution channel, WAS line, then DAFd
RAS channel
Appleton, WI 22 ( t . o) Fine bubble/coarsebubble MLb distributionchannel Altemative WAS wasting system to
DAF
President Street, 27 ( t . 2) Fine bubble/Ml channels Aeration basin Combined with WAS and primary
Savannah,GA sludge prior to DAF
South Cobb County, GA 40 ( 1. 8) Fine bubble/coarsebubble AB" effluent collection
channel
Metro plant, 80 (3.6) Fine bubbleicoarse bubble RAS re-aeration basin, end Combined with WAS prior to DAF
St. Paul, MN of aeration basin passes
Haskell R. Street plant, 29 ( 1. 3) Fine bubble/coarsebubble RAS channel Combined with WAS prior to DAF
El Paso, TX
Utoy Creek, Atlanta, GA 45 (2.0) Fine bubble/coarsebubble MLb distribution channel Combined with WAS ahead of
thickening centrifuges

o Maximum month flow.

b Mixed liquor.

' Aeration basin.

d Dissolved flotation.

Source:From Parker,D.S. et al. (2003), Water Environ. Res.,75,83. With permission.

154 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
-r
-l
^l
l'l
YV
T-
I
I Direction of
50-250mm
variableopeningfor
scum ( a)
FICURE 5.29 Nocardioformfoam control devicefor surfacechlorinationat the 23rd Avenuewastewatertreatmentplant in Phoenix.
AZ: Gl side view and tb) top view.
6. Chlorination
RAS chlorination is not very effective for controlling
nocardioformsin activatedsludge becausethe nocardio-
form filaments are largely contained inside the activated
sludge flocs. Thus, they are inaccessibleto the chlorine
unless a high enough dose (actually an overdose) is
applied to break up the flocs. Such dosesare not recom-
mended becausethey degradeeffluent quality and dimin-
ish treatmentcapacity.Some benefit from RAS chlorina-
tion in controlling nocardioformscan be achievedin a
system with an aerationbasin that has subsurfacewith-
drawal becausethis generatesfree-floatingnocardioform
filamentsthat are accessibleto the chlorine.
A far more effective use of chlorine in nocardioform
control is to apply a chlorinesolutionas a fine spray(mist)
directly to the aerationbasin surface.This approachhas
been successfulat severalplantsin the U.S. including the
23rd Avenue plant in Phoenix; Stamford, CT; Ocean
County,NJ; Trenton,NJ. (Albertson,1991);and SanJose,
CA (Ekster and Jenkins, 1999).At the 23rd Avenueplant
in Phoenix,sprayhoods (Figure 5.29) were placedacross
the end of the third pass of the four-pass aeration basin.
As the foam entered the hood with the mixed liquor flow,
it was sprayed at a rate of approximately l0 L/min with
a chlorine solution (from a gas chlorinator) containing 2 to
3 mg Cl,rlL. Using a chlorine dose in the range 0.5 to
1.0 mg Clrll- basedon the wastewaterflow, nocardioform
foam was eliminated on severaloccasionswithin I to 2 d
without any effluent quality deterioration or loss of treat-
ment efficiency.
At the San Joseplant, a fine mist spray of NaOCI (3 g
Clrll-) solution was applied through a shroudedspray bar
to the surfaceof one aeratedcompartmentof a 4-compart-
ments-in-series,step-feedBNR activatedsludge system.
At dosesof 0.5 to 1.0mg Clrll- (basedon wastewaterflow),
this systemcontrolled nocardioform foam without altering
bulk mixed liquor nocardioform organism levels, suggest-
ing that it only kills nocardioforms at the surfacesof the
Heavily
chlorinated
water spray
(Clt=2-3 t11',
mixed liquor flow
(b)
basins.This method of nocardioform organism control did
not compromisethe quality of the BNR effluent in any way.
It may be possibleto enhancethe effectivenessof a
chlorine surface spray by installing fine bubble aeration
to enhance the nocardioform foaming just prior to or
within the chlorine surface spray system. When using
surface spray chlorination, it is important to avoid dis-
persal of the sprayed(misted) chlorine solution because
ofits hazardousand corrosivenature.Further,when using
a NaOCI solution spray,flushing with water is necessary,
especiallyprior to shut-down,to preventblockageof solu-
tion lines and sprayswith the sparingly soluble precipitates
that can form by interactionof the alkaline NaOCI with
water and the atmosphere.
7, Cationic Polymer Addition
In the discussionof the effect of reactorwithdrawal con-
figuration on nocardioform growth form, it was shown that
in pure cultures of G. amarae, an overflow outlet tended
to produce clumped growth while a subsurfacewithdrawal
configuration tendedto producedispersedfilaments.These
sameeffectscan be observedin activatedsludgecontaining
nocardioforms. Indeed the microscopic observation of
free-floating nocardioform filaments in a mixed liquor
samplecan be used as an indication that the aerationbasin
has a subsurfacewithdrawal and that foam trapping and
an accumulationof foam will occur on the aerationbasin.
Because the hydrophobic nocardioform organism
surface causes foaming, the presence of free-floating
nocardioforms will enhancefoaming over the levels that
would be produced by nocardioforms buried inside acti-
vated sludge flocs (Narayanan,2002). A subsurfaceaera-
tion basin outlet producesfree-floating nocardioforms that
foam more efficiently. The foam builds up on the basin
surfacebecauseit is trappedby the subsurfaceoutlet. Shao
etal. (1997\ found that the addition of small amounts
(0.5 mg/L basedon wastewaterflow) of a cationic poly-
acrylamidepolymer (e.g., Polydyne Inc. Clarifloc-C34l,
A c t iv at edS lud g eF o a mi n ga n d C o n tro l 155

100 IOO (,
6

Eqo
a"
4

' 6t, 80 e,

?70
!60 60*

E50
E
b40 40 .2
d 90
dln

o lt) 20E
E
ri '" a

oz
Jan Jan Jan Jan Feb Feb
9 t6 30 6 t- 1

Date,1995

FICURE 5.30 Effect of cationic polymer additionon nocardioformconcentrationand aerationbasin foam coverage,Terminal Island
Plant, Los Angeles,CA. (From Shao,Y.J. et al. (199'7),Water Environ Res.,69,25. With permission.)

aminomethylatepolyacrylamide, low charge density,

molecular weight 20 to 30 x 106)to the aeration basin tr
eliminatednocardioformfoam. Figure 5.30 showsthat the
reductionof aerationbasinfoam coveragewas not accom-
paniedby a proportionaldecreasein nocardioformorgan-
ism counts.Shaoet al. (1997) postulatedthat the polymer EX

flocculates the free-floating nocardioforms back into the

oE
activatedsludge flocs, thereby decreasingthe amount of 'E-

hydrophobic surface available for attachment to air bub- o:

bles and thencedecreasing foam formation.

By eliminating the nocardioform foam, the polymer
addition reincorporatesthe nocardioformorganismscon-
centratedin the foam back into the mixed liquor. Here
they are lesslikely to causefoaming and are accessibleto FIGURE5.31 Effecton aerationbasinfoamcoverage
wasting in WAS so that they can be eliminatedfrom the mg/Lcationicpolymerat activated
activated sludge at the samerate as other activated sludge
microorganisms.
Polymer addition to control nocardioform foam has
been practiced at severaltreatmentfacilities subsequent
to the initial trials at the Terminal Island plant in Los
Angeles. For example, nocardioform foam on the aeration
basins at the plant in Frankenmuth, MI was reduced from
75%osurface coverage (2 in. deep) to a l07o coverage
(0.25 in. deep) by the addition of I mg/L of polymer for
4l d (Figure5.31).
The polymer can be dosed either to an RAS wet well
or to a mixed liquor channel.Polymer dosing periods of 3
to 10 d were usedat San Joseto control nocardioform foam
on three occasions.During the first test period (July 1996),
aerationbasin foam and scum levels in the secondaryclar-
ifier feed channelwere noticeablyreducedafter I d ofinter-
mittent polymer dosing at 0.5 mg/L. After 2 to 3 d of
continuouspolymer dosing at 0.5 mg/L, the foam and scum
virtually disappeared.During the secondtest period (Janu-
15 20 25 30 35 40 45
Time,d
of dosingI
sludgeplantin Frankenmuth, MI.
ary through March 1997), a polymer dose of 0.2 mglL
controlled foaming most of the time but was inadequateto
completely eliminate it. Increasingthe polymer dose to 0.4
to 0.5 mg/L controlled the foam. During the third test period
(April 1997),foam and scum were controlled rapidly using
a polymer doseof 0.35 mg/L (Eksterand Jenkins,1999).
One of us (DJ) was told an interestinganecdoteduring
a discussionof the use of polymer for nocardioformfoam
control. On hearingaboutthis methodand our hypothesis
for how it works, a treatment plant operator replied, "So
that's why my foam goes away when I overdosepolymer
to my belt presses!" [Languageparaphrasedto remove
certainAnglo-Saxonadjectivesand verbs.l This very per-
ceptive operator figured out that the type of polymer used
for sludgethickening by gravity belts, belt filter presses,
centrifuges, etc. is the same type that works for nocardio-
form foam control and that an overdoseof polymer to his
belt pressesresulted in a recycle of polymer to the aeration
156 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems
'70
Averase MCRT 1996= 5.1 d
60 AverageMCRT 1997= 613d
50
tr
.. +v
u^^
>JU
Nov 16 Dec17 Jan16 Feb15 Mar 16
DateE
FIGURE 5.32 Comparisonof manual control (1996) and auto-
matic control (1997) of MCRT at the SanJose/Santa
Clara plant.
(From Ekster,A. and Jenkins,D. (1999), Proc. Water Environ.
Fed. Conf. WEFTEC '99. With permission.)
basin in the filtrate and that this was what eliminated the
aeration basin foam.
B. Automatic MCRT Control
Recentwork at the San JoseBNR plant indicatesthat on-
line instrumentalmeasurements of activatedsludgeMLSS
and RAS SS togetherwith automaticcontrol of wasting
may be betterthan manual(laboratory)SS measurements
and manual activatedsludge wasting for preventingacti-
vated sludge foaming. Online instrumental MLSS and
RAS SS analyseswith the datausedfor automatedMCRT
control eliminated nocardioform foaming from the San
Jose plant and the need to use surfacemist chlorination
or polymer addition for foaming control.
It is possiblethat MCRT control with automatic sludge
wastingandcontinuousTSS analysesallows for much less
variationin discreteMCRT values.This point is illustrated
by the MCRT datain Figure 5.32. Note how the manually
controlled sludgewasting basedon laboratoryTSS anal-
yses (1996 data) includes many incidents of very high
MCRT. It is possible that such incidents encouragethe
growth of nocardioforms that then are retained in the
activated sludge system by surface foam trapping even
when MCRT is lowered. It was possible to operate in a
nocardioform-free condition at a higher average MCRT
(6.3 d) than with manual activatedsludge wasting based
on laboratoryTSS analyses(5.1 d).
IV. MICROTHRIX PARVICELLA
A. FacronsArrrcrrrucM. penwcruACRowrH
1. General
Excessivegrowth of M. parvicella in activatedsludge can
causeboth bulking and foaming. Like nocardioform organ-
isms, M. parvicella organismsappearto have hydrophobic
cell walls and they can stabilize brown, high-solids foam
on activated sludge. BecauseM. panticella filaments are
much longer than nocardioform filaments, they can
decreaseactivatedsludgesettling ratesby bridging between
flocs. The morphology of M. panicella is stch that very
large populations are needed to cause severebulking. A
curved and coiled filament usually grows throughout and
wraps around the activated sludge flocs, rather than pro-
truding rigidly from the floc surfacesinto the bulk solution.
M. parvicella was first describedby van Veen (1973)
and first isolated by Slijkhuis (1983a and 1983b). Like
nocardioforms, it is strongly Gram-positive and contains
Neisser-positivegranules.
2. Substrates
The original isolate of M. parvicella required long chain
fatty acids(e.g.,oleic acid) or theirTween estersfor growth.
More recentstudies(Wanner,1994;Holmstromet al.,1996:
Tandoi et al.,1997) show that simple carbon substratessuch
as glucose,acetate,and ethanol can support the growth of
M. parvicella. Because M. parvicella-dominated sludge
formed in a predenitrification activatedsludge system fed
with substratessuch as glucose and ethanol, Wanner and
Grau (1988) suggestedthat it could denitrify and form stor-
age products from simple carbon substrates.
Neilsen et al. (2001) showed that M. parvicella can
take up and store long chain fatty acids such as oleic acid
under anoxic and anaerobicconditions.They suggestthat
a surface-associatedlipase breaks down the oleic acid at
the hydrophobic filament surface and facilitates its trans-
port into the cell and storageas neutral lipids. Neilsen et al.
(2001) postulatedthat theseabilities give M. parvicella a
competitive advantagein anaerobicand anoxic conditions
where hydrolysis rates of particulate organics by the acti-
vated sludge community as a whole are much lower than
under aerobicconditions(Dold and Marais, 1986).Under
aerobic conditions, floc-forming microorganisms can
metabolizelipid substratessuch as oleic acid. Nielsen et al.
(2001) suggestthat this ability allows floc formers to out-
compete M.parvicella in well mixed, completely aerated
activated sludge and in aerobic selector systems.
3. OperatingConditions
M. parvicella is not currently a common dominant fila-
mentous organism in activated sludge in the U.S.
(Table2.1). We have observedit mostly in high MCRT
plants in upper tier states,(Michigan, Wisconsin,Minne-
sota,Wyoming, Montana, and Colorado) especially during
the winter. In many other parts of the world (Northern and
SouthernEurope, South Africa) it is by far the most com-
mon dominant filamentous organism in activated sludge
systems. This is likely due to the presence of a higher
ActivatedSludgeFoamingand Control 157

percentageof high MCRT BNR plants in these areasthan
in the U.S. While the specific causes for excessive
M. parvicella growth in activated sludge are not com-
pletely understood,the four most commonly cited condi-
tions associatedwith its srowth in activated sludge are:
. High MCRT
. Low DO
. Low temperature
. Presence of anoxic. anaerobic. and intermit-
tently aeratedzones such as those used in BNR
plants and presentin mechanically aeratedaer-
ation basins
As high MCRT BNR plants become more common
in the U.S., the incidenceof M. parvicella is cerrainto
increase.
Wentzel (1992) reported that at the five-stageBarden-
pho JohannesburgNorthern plant (South Africa), M. par-
vicella was the dominantfilamentousorganismin the win-
ter. In the summer, Type 0092 became dominant.
Knoop and Kunst (1998), in pilot plant and full-scale
experimentson municipal wastewatertreatment plants
containinganoxic and/or anaerobiczones,found that the
best growth of M. parvicella occurredat <12 to l5oC and
with F/M values of 30.1 kg BOD./kg MLSS/d. Under
these conditions, M. parvicella filaments were 200 to
500 pm long and SVI values were as high as 490 mLlg
(average, 250 mLlg). At >20"C and the same F/M, the
M. parvicella filaments fragmentedinto 30- to 80-pm long
pieces, then gradually disappearedfrom the activated
sludge.The filament fragmentationwas accompaniedby
an SVI decreaseto approximately 150 ml-/g. When the
F/M was increasedto >O.2kg BODr/kg MLSS/d, M. par-
vicella was gradually eliminated from the activatedsludge
even at <12 to 15oC.Before disappearingfrom the acti-
vated sludge, the M. parvicellafilaments formed rope-like
bundles that did not cause as much interference with set-
tling and SVI valuesdecreasedto <150 ml/g.
Mamais et al. (1998) investigatedthe effects of reactor
mixing conditions and the presenceof anoxic zones on
bulking due to M. parvicella. Using pilot-scale activated
sludgeoperatedat MCRT = 18 d, F/M = 0.2 kg COD/kg
VSS/d, and temperature= 14 to 20oC on municipal waste-
water, the following conditions were examined:
. Completely aerobic plug flow
. Plug flow predenitrification (an anoxic selector
followed by a plug flow aerobic basin with
internal recycle from the end of the aerobic
basin to the anoxic zone)
. Completely aerobic,completely mixed
. Completelymixed, intermittentlyaerated,nitri-
fi cation-denitrifi cation
All reactors had foam-trapping features in their aera-
tion basinsand secondaryclarifiers.The best settlingwas
obtained from systems with plug flow aerobic basins
(Figure 5.33). The two sludges contained very few
M. parvicella filaments. The activated sludge from the
completelymixed nitrification-denitrification systemwith
CSTR basinsshowedthe poorestsettling and M. parvice'
lla was dominant. The completely aerobic, completely
mixed system settled better than the completely mixed,
nitrification-denitrificationsystembut poorerthanthe sys-
tems with plug flow aerationbasins.
Gabb and coworkers (1991, 1997a, 199'lb) worked
with continuously fed and intermittently fed laboratory
activated sludge systems with a range of aerobic and

120

^O eAa
:>
F
o€ *
og
^MAMMaaaeS
D
euI eaaaa^aAAr ,otooo*o?oooo
}E D Ja"
^oo
> .:
u)=
o
o Aerobic plug-flow
!E o
*r8**
FO
na ^^"- o PTeNDN plug-flow
40 $a
qP X
vu a Aerobic completely mixed
o-; ^o
-F x Intermittent aern, NDN
,' completely mixed
f oo"',,"u.?*
0 50 100 150 200 250 300 350 400 450 500
DSVI, ml-/g

FICURE 5.33 Effect of unaeratedzones and aeration basin configuration on the settling of activated sludge dominated by
M. parvicella. (From Mamais, D. and Jenkins,D. (1992), Water Sci. Technol.,26:5-6,955. With permission.)
158 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
anoxic conditions treating settled municipal wastewater.
MCRT valueswere 20 to 30 d, F/M was 0.28 to 0.31 kg
COD/kg VSS/d, and temperaturewas 20'C. Completely
aerobic systems did not bulk and did not support the
growth of low F/M filamentous organisms (including
M. parvicella) whether or not they had selectors.
Continuously fed, intermittently aerated activated
sludge (simulating an oxidation ditch type of aeration
basin) had high SVIs due to proliferation of Type 0092
and M. parvicella. The installation of an aerobic selector
aheadof the intermittently aeratedbasin did not eliminate
thesefllamentousorganisms.When the aerationbasinwas
continuouslyaerated(i.e., the units were completelyaer-
obic), bulking due to Type 0092 and M. parvicella was
eliminated.
4. Control
The growth of M. parvicella tn activatedsludge can be
reducedby:
. ReducingMCRT to as low a value as possible
consistentwith achievingtreatmentobjectives
. Providing plug flow basinsin both anoxic and
aerobiczones
. Avoiding intermittentaerationor zonesof low
DO (<1 to 2 mglL) in aerobiczones,especially
at the point where mixed liquor from an anaer-
obic or anoxic zone entersan aerobiczone
. Eliminating foam trapping features
. A full-scale aerobicselectorwas subsequentlyincorporated
Addng a polyaluminum chloride-basedfloccu-
lant
Recent work in The Netherlandsand Japan (Eikel-
boom, 1997;Hwang et al., 1998;Roels et al., 2002) has
shown that M. parvicella bulking and foaming was con-
trolled by the addition of polyaluminum chloride
(PAX-14, Kemwater B.V., Rozenburg,The Netherlands)
to the aeration basin. Dose rates for low MCRT activated
sludge systemswere functions of MCRT (60/MCRT =
PAX-14 dose,g as Al/kg MLSS). Using this criterion and
an extended aeration activated sludge system with
MCRT =20 d, HRT = I d, and MLSS = 3000 mg/L would
resultin an aerationbasininfluentPAX- 14 doseof approx-
imately 0.5 mg Al/L. That amount is on the same order
of magnitudethat is effective for controlling nocardioform
foaming with cationic polyelectrolyte.
Roels et al. (2002) report that control is achievedafter
about I to 2 weeksof PAX-14 dosing.It was also reported
that PAX-14 addition seemsto affect M. parvicella stain-
ing propertiesand filament morphology, suggestingthat
someinhibitory toxic effectin additionto flocculationmay
occur. PAX-14 addition appeared to have no effect on
nocardioforms or N. limicola.
Pr oblem s
5. Case Histories
The following two examples illustrate how the presence
of alternating anoxic and aerobic zones in an aeration
basin stimulatesthe growth of M. parvicella and how their
elimination and the use of an aerobic selector controls
M. parvicella.
a. Upper Occoquan Sewage Authority (UOSA), VA
The UOSA Regional Water Reclamation Plant is a
2'7-N4GD(1.2 m3/s) facility utilizing preliminary treat-
ment, primary treatment,and single sludge nitrification.
Extensivephysical/chemicaltreatmentof the nitrified sec-
ondary effluent is provided to allow discharge of the
treated effluent to a drinking water reservoir. Effluent dis-
charge standardsand plant reliability and redundancy
requirements are extremely stringent, resulting in a con-
servativedesignfor the biological treatmentfacilities with
a relatively long hydraulic residencetime, low organic
loading rate, and low secondaryclarifier overflow rate.
The biological treatment plant was upgraded and
expandedfrom an existing completely mixed activated
sludge system that historically experiencedseverebulking
problems with SVIs as high as 600 ml-/g that restricted
treatment capacity. Poor settleability resulted in reduced
MLSS concentrations,reducedMCRT, andreducedcapacity
to nitrify. Periodic identification of the filamentous organ-
isms presentindicated that M. panicella was typically pre-
dominant,especiallyduring cold weather.A bench-scale
evaluation was conducted of an aerobic selector (DO
)_2mglL) for conection of the filamentousbulking problem.
into the design of the expandedactivatedsludge system.
Figure 5.34 is a schematicof the expanded,upgraded
full-scale, system. The selector consists of an aerated
channel(length:widthratio of 5:l) with a hydraulic resi-
dencetime of 11 min basedon influent flow. Dye testing
indicated that the hydraulic flow pattern in the selector
could be approximatedas three equal-sizedcompletely
mixed tanks in series.Following the selectorare two sets
of activatedsludgeaerationbasinsin series.The first set
comprises newly added diffused air basins required to
Primary effluent
RAS
Mixed 5.1".1o.
liquor
To secondary
claritiers
Diffused
air
Aeration basin
FIGURE 5.34 Aeration basin and selectorsystem,UOSA, VA.
(From Daigger,G.T. and Nicholson,G.A. (1990), Res.J. Water
Pollut. Control Fed.. 62. 676. With oermission.)
ActivatedSludgeFoamingand Control
700
600 Selector
I
500 Voperatlonal
il
) +oo
<---+ difficulties
F 3oo
O
200
100
0
t98'r 1988
FIGURE 5.35 Effect of selectoroperationon SVI at UOSA, VA. (From Daigger, G.T. and Nicholson, G.A. (1990), Res.J. Water
Pollut. Control Fed., 62,676. With permission.)
accomplishthe plant expansionand the secondcomprises
the existing completely mixed basins using slow speed,
surface mechanical aerators.The volumes of the new dif-
fused air and existing mechanical aeration basins are
approximately the same.
Excellent sludge settling,as indicatedby an average
SVI of 14 mLlg, was obtained with the expandedand
upgradedsecondarytreatmentsystemwhile operatingin
excessof designorganic loading and with periodsduring
which hydraulicloadingwas equalto the maximum month
design value.Figure 5.35 presents4 y of data illustrating
the impact of the aerobicselectoron sludge settleability.
Before March 1988,the existingcompletelymixed basins
using surfacemechanicalaerationwere in service.Severe
sludgebulking (and some foaming) problemswere expe-
riencedregularly,and SVIs typically were>150 ml-/g and,
on occasion,as high as 600 ml/g. The predominantfila-
ment was M. parvicella. In March 1988,the aerobicselec-
tor and diffused air aerationbasinswere brought on line
and the existing mechanicallyaeratedbasinswere taken
off line. The aeration basin hydraulic residence time
remained about the same becausethe volumes of the
mechanicallyaeratedand diffusedair aerationbasinswere
about the same. The result of the change in reactor con-
figuration was a rapid decreasein SVI (Figure 5.35) as
M. parvicella was eliminated from the system.The SVIs at
UOSA haveremainedlow since installation of the selector,
exceptfor a brief period in late 1988 when construction-
relatedproblemsresultedin unusualoperatingconditions.
Operatingconditionswithin the selectorwere charac-
terized in February 1989. Direct measurements indicated
soluble BOD, removalsof approximately 6OVoand soluble
COD removals of approximately 457o through the selec-
tor. Oxygen requirements exerted in the selector were
determinedby (1) measurementof the oxygen uptake rates
of samples withdrawn from the selector and (2) direct
calculation from measuredair flow rates using measured
DO concentrationsand oxygen transfer efficiencies deter-
mined by using an off-gas analyzer.
159
Construction
7
y' related
1989 I 990 l99l
Year
Both techniques produced similar results and indi-
cated an oxygen requirement in the selector of approxi-
mately 0. 1 mg Orlmg soluble BOD, removed.The DO
concentrationsin the selectorhave been maintainedcon-
sistently above2 mg/L, suggestingthat DO doesnot limit
the rate of oxidation of carbonaceousmatter. This rela-
tively low oxygen requirement suggeststhat storage,
rather than oxidation, is the predominant fate of removed
soluble organic matter in the selector. The F/M for the
entire selector was 4.9 kg BODr/kg MLSS/d while the
F/M, for the first selector compartment was 14.8 kg
BODr/kg MLSS/d.
Becausetwo changeswere made when the UOSA
plant was upgraded- installationof the aerobicselector
and addition of a diffused air aerationbasin upstreamof
the existing mechanically-aerated aerationbasin - it is
not possibleto determinethe relativecontributionsof each
modification to improving the sludgesettling characteristics
and controlling M. parvicella. Howevertheseobservations
are consistentwith the findings of Mamais et al. (1998)
and Gabb et al. (1991,1997a,1997b)that M. parvicella
growth is controlled by completely aerobicselectorsand
aeration basinsbut not by aeration basins that contain low
DO zones.The following case study provides additional
insight.
b. Northside Wastewater Treatment Plant,
Tulsa, OK
This plant provides preliminary treatment, primary treat-
ment, and secondary treatment using the complete-mix,
air-activated sludge processwith mechanical surface aer-
ators. Initially the plant provided 28 MGD (1.2 m3/s)of
capacity using two aeration basins. Nitrification occurred
periodically, but the capability to achieve consistentnitri-
fication was limited by chronic filamentous bulking prob-
lems with SVI values as high as 500 ml-/g for periods of
several consecutive weeks. M. parvicella was often the
predominant fi lamentous microorganism causing bulking
durine cold weather.
160 M anua lo n C a u s e sa n d C o n tro lo f A c ti v atedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Surface
mechanical
aerators(Typ)
Primary
effluent
Return
activated
sludge
Diffused air
Aerationbasin
FICURE 5.36 Northside plant aerationbasin with aerobic selector,Tulsa, OK. (From Daigger, G.T. and Nicholson, G.A. (1990),
Res.J. WaterPollut. Control Fed., 62.676. With permission.)
Expansion of the plant required reliable, consistent
year-round nitrification. Two additional aeration basins
with essentiallythe same configurationsas the existing
basins were to be provided. To addressthe persistent
sludge bulking problems, an aerobic selectorwas to be
providedalong with the two new aerationbasins.The new
basins were square and equipped with nine slow speed
mechanicalsurfaceaeratorsand influent piping networks
that distributedthe influent throughout the basins.Each
basin had an aerobic selector consisting of a two-pass
channel with a length:width ratio of 17:1 that received
primary effluent and RAS (Figure 5.36). The selector
HRT, basedon a design flow of 14 MGD (0.61m3/s)per
basin.was l6 min: the HRT in the aerationbasinwas 5.5 h.
The new aerationbasinswere startedup in January1988
and, after a 6-mo acclimationperiod, the performanceof an
aerationbasin with an aerobicselectorwas comparedto the
performanceof an existing aerationbasin with no selector.
Contrary to the experienceat UOSA, the aerobic selector
system settling characteristicswere no better than those in
the systemwithout an aerobicselectorover an I 1-mo period
when the two systemswere operatedwith the sameprocess
loadings and operating parameters.M. parvicella was the
predominantfi lamentousorganism causingbulking.
In the aerobic selector system, the SVI averaged
152 mLlg. In the UOSA aerobicselectorsystem,the aver-
age SVI was 72 ml/g. The overall F/M in the Nonhside
selectorwas 3.2 kg BOD.ikg MLSS/d- somewhatlower
than the value of 4.9 kg BOD./kg MLSS, d at UOSA. The
very high length:width ratio of the Northside selector
makesit likely that its initial contactzone F/M is compa-
rable to or greater than that at UOSA. The Northside
selector removed the same fraction of soluble BOD.
(607o)as the UOSA selectoralthoughoxygen uptakeand
airflow rate data suggestedthat more of the soluble BOD,
was oxidized rather than stored.
P roblem s
A comparison of UOSA and the Tulsa Northside aer-
obic selectorperformancessuggeststhat an aerobicselec-
tor alone is not capable of controlling sludge bulking
caused by M. parvicella. The configuration of the main
aeration basin also seems to affect the occurrence of
M. parvicella. A main aeration basin with uniform aera-
tion (such as a floor covered with fine bubble diffusers)
seemsto provide greater control of M. parvicella growth
than a basin with point sourceaerationdevices(such as
mechanicalsurfaceaerators).
V. ANAEROBIC
DICESTER
FOAMING
Various aspectsof anaerobic digester foaming causedby
nocardioformorganismshavebeendiscussedin this chap-
ter. In summary:
. Anaerobic digester foaming problems due to
nocardioforms and M. parvicella originate in
aerationbasins(where theseorganismsgrow).
The foam-causing microorganismsare intro-
duced into the anaerobicdigesterin the WAS.
. Foaming due to these filamentous organisms
can occur in anaerobic digesters before nui-
sancefoams appearon aerationbasinsbecause
the filamentous organism concentrationper unit
volume can be higher in an anaerobic digester
than in the mixed liquor becausethe SS content
of the WAS fed to the digester is higher than
the MLSS, especiallysince the WAS feed has
usually been thickened.
. Serious operating and equipment problems can
be causedby nocardioform foaming in anaero-
bic digesters.Figure 5.37 shows the resultsof
liquid phase percent total solids and tempera-
ture profiles of the contents of an anaerobic
digester at the San Francisco South East plant.
 ActivatedSludgeFoamingand Control
Cover
6.3 84
5 .5 9t
4.1 94
5.0 89
4.5 93
Total solids,To Temperature,oC
FIGURE 5.37 Nocardioformfoam effectson percenttotal solids
and temperature profiles in an anaerobic digester, South East
plant, San Francisco,CA.
Note the large fraction of digestervolume occu-
pied by nocardioform-induced foam, inversion
ofTS concentrations(higher on the top than on
the bottom of the digester), and temperature
decreaseof the liquid in the foam layers.
Nocardioforms and M. parvicella both lose
their Gram-positive staining properties and
their Neisser-positivegranulesduring anaero-
bic digestion. Nocardioform fi lamentsfragment
and the long coiled M. parvicella filaments
break into shorter, thicker filaments. Generally
these changesmake it more difficult to micro-
s c opic a l l y re c o g n i z e n o c a rd i o fo rms and
M. parvicella in anaerobic digesters than in
activated sludge. For this reason, Hernandez
et al. (1994) developed an immunofluorescent
antibody staining method for G. amarae that
made it possible to count these organismsin
anaerobically digesting sludge. Using this
method and a dehydrogenaseenzyme method
for viability determination, Hernandezand Jen-
kins (1994) showedthat the viability of G. ama-
r ae de c re a s e d o n l y v e ry s l o w l y duri ng
anaerobicdigestion(decayrate = 0.04/d).Thus,
recycle streams from digested sludge process-
ing (e.g., centrate, filtrate, and supernatant)
could seed the activated sludge plant with
noc ar d i o fo rms . T h e fo a m i n g p ro p erti es
decreasedeven more slowly than the viability
during anaerobic digestion, suggestingthat the
tendency to foam is retained by nonviable
G. amarae cells or cell walls.
The amount of foam produced by nocardio-
forms in anaerobic digesters is greatly influ-
enc ed b y d i g e s te r mi x i n g m e th o d s and
geometry. The production of CO, and CH, by
161
0.4
Fine bubble gas mixing
.90n1
"'"
€
,t
J
at
0.2
-
d
Coarse bubble gas mixing
F o.t
ri
Mechanicalmixing
2468101214
Time.min.
FIGURE5.38 Foamaccumulation ratein experimentalanaer-
obic digestersas a function of mixing technique.(From van
Niekerk,A.M. et al. (1987),J. WaterPollut.ControlFed.,60,
100.With permission.)
the digestion processcan produce foam without
mixing. This is most evident immediately after
feeding the digesterwhen gas productionusu-
ally increases.Figure 5.38 showsthat fine bub-
ble gas mixing devices(unconfinedgas mixers,
gas lances,etc.) producethe greatestamountof
foam; coarsebubble types of mixers produce
less foam; external mechanical mixers and
pump mixing have the least foam-producing
potential.Becausethey have much smaller top
liquid surfaces, egg-shaped digesters tend to
allow smaller accumulations of foam than
cylindrical digesters.
. The best method for controlling nocardioform
or M. parvicella foaming in anaerobicdigesters
is to eliminatecausativefilamentousorganisms
from the activated sludge system so that they
never reach the anaerobicdigesters.The use of
mechanicalmixing, external pump mixing. or
coarse bubble mixing rather than fine bubble
mixing will reducefoaming potential. Westlund
et al. (1996, 1998a, 1998b)found that a mixer
placed in the gas space above the digesting
sludge can be used to collapse foam. Mixers
that spray sludge onto the surfaceofthe digest-
ing sludge can help collapse foam.
 B iblio g r a p h ya n d R e fe re n ce s
The historian, essentially,wants more documents than he
can really use; the dramatist only wants more liberties
than he can take.
The Aspern Papers, Henry James,1843-1916
In addition to references cited in text, this section lists
additional literature about the causes and control of acti-
vated sludge solids separation problems.
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Albagnac,G. and Morfaux, J.N. (1980), Traitabiliti6 Compar6e
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Albertson,O.E. (1991),personalcommunication.
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Sci.Technol.,26:34, 461.
Al-Diwamy, L.J. and Cross, T. (1978), Ecological Studies of
Nocardia Foams and Other Actinomycetes in Aquatic
Habitats,in Nocardia and Streptomyce^s; Modarski, M.
et al., Eds., Gustav FischerVerlag, Stuttgart,p. 153.
Allen, L.A. (1944), The Bacteriology of Activated Sludge,
J.Hys., 43,424.
Amann, R., Snaidr,J., Wagner,M., Ludwig, W., and Schleifer,
K.H. (1996), In Situ Visualization of High Genetic
Diversity in a Natural Microbial Community,J. Bacte-
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Andreasen,K. and Nielsen,P.H.( 1998),In Situ Characterization
of Substrate Uptake by Microthrix parvicella Using
Microautoradiography,Water Sci. Technol., 37:4-5, 19.
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dation of Sewage without the Aid of Filters, J. .Soc.
Chem.Industr.,33, 523.
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without the Aid of Filters, J. Soc. Chem. Industr., 33,
lt2.
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Selector Removal on Biomass Composition, Res. J.
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tous Eikelboom Type 02lN Bacteria and Descriptions
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AIV Working Group 2.6.1 (1989),Report:Preventionand Con-
trol of Bulking Sludge and Scum, Korr. Abwass., 36,
165.
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(1953), Protozoa as Indicators in Activated Sludge
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Pollut. Control, 77, 103.
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Treatment Plants, Appl. Microbiol., 3, l'73.
Beebe,R.D. (1983),personalcommuniation.
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Bulking at the San Jose/SantaClara Water Pollution
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the California Water Pollution Control Association,
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Beebe,R.D., Jenkins,D., and Daigger,G.T. (1982),Activated
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Nocardioform Actinomyceten in Modellklaranliigen,
Korr. Abwass.,36, 152.
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(2002),FilamentousChloroflexi GreenNon-Sulfur Bac-
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163
164
M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Blackall, L.L., Parlett, J.H., Hayward, A.C., Minnikin, D.E.,
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(1996),TowardsUnderstandingthe Taxonomyof Some
of the FilamentousBacteriaCausingBulking and Foam-
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Blackall, L.L., Seviour,E.M., Cunningham,M.A., Seviour,R.J.,
and Hugenholtz, P. (1994 ), Microthrix parvicella is a
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Blackall, L.L., Tandoi, V., and Jenkins,D. (1991), Continuous
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on FilamentousMicroorganismsResponsiblefor Bulk-
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Pr oblem s
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Buchanan,R.E. and Gibbons,N., Eds. (1974), Bergey'sManual
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Cairns,W.L., Cooper,D.G.,Zajic, J.E.,Wood, J.M., and Losaric,
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Curds,C.R.,Cockburn,A., andVandyke,J.M. (l968), An Exper-
imental Study of the Role of the Ciliated Protozoa in
the ActivatedSludgeProcess,WaterPollut. Control, 67,
312.
Curds, C.R. (1975), Protozoa,in Ecological AspectsoJ Used-
Water Treatment: The Organisms and Their Ecoktgr*,
Curds,C.R. and Hawkes,H.A., Eds.,AcademicPress,
New York. chap. 5.
Curtis, E.J.C, (1969), SewageFungus: Its Nature and Effects,
Wuter Res.,3,28tJ.
Cyrus,Z. and Sladka,A. (1970), SeveralInterestingOrganisms
Presentin Activated Sludge,Hydrobiologia, 35, 383.
Daigger,G.T. (1995), Developmentof RefinedClarifier Operat-
ing Diagrams Using Updated Settling Characteristics
Database,Water Environ. Res.,6'7,95.
Daigger, G.T. and Buttz, J.A. (1988), Upgrading Wastewater
TreatmentPlants, 2nd ed.,TechnomicsPublishers,Lan-
caster.PA.
Daigger,G.T. and Nicholson,G.A. ( 1990),Performanceof Four
Full-Scale Nitrifying Wastewater Treatment Plants
Incorporating Selectors, Res. J. Water Pollut. Control
Fed.,62,676.
Daigger, G.T. and Roper, R.E., Jr. (1985), The Relationship
between SVI and Activated Sludge Settling Character-
istics,-/. WaterPollut. Control Fed., 5'7,859.
Daigger, G.T., Robbins, M.H., Jr., and Marshall, B.R. (1985),
The Design of a Selector to Control Low F/M Filamen-
tous Bulking, J. WaterPollut. Control Fed., 57,220.
1 66
M anua lo n C a u s e sa n d C o n tro lo f A c ti v atedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Daigger, G.T., Waltrip, G.D., Romm, E.D., and Morales, L.A.
(1988), Enhanced SecondaryTreatment Incorporating
Biological Nutrient Removal J. Water Pollut. Control
Fed ..6O.1833.
Das, D., Keinath,T.M., Parker,D.S., and Wahlberg,E.J. (1993),
Floc Breakup inActivated Sludge Plants, Water Environ.
Res..65. 138.
De, L.G. and Solbe,J.F.(1975),Annelida, in EcologicalAspects
of Used-Water Treatment: The Organisms and Their
Ecology,Curds,C.R. and Hawkes,H.A., Eds.,Academic
Press,New York, chap. 8.
Deakyne,C.H.W., Patal,M.A., and Krichten, D.l. (1983),Dem-
onstration of Biological Phosphorus Removal by the
A/O Process at 70-MGD Patapsco Wastewater Treat-
ment Plant, Whitman RequardtAssoc.,Baltimore,MD.
Design of Municipal WastewaterTreatmentPlants (1998), Man-
ual of Practice 8, Water Environment Federation, AIex-
andria, VA.
Dhaliwal, B.S. (1979), Nocardia amarae and Activated Sludge
Foaming,J. WaterPollut. Control Fed., 51,344.
Dias, F.F.and Bhat, J.V. (1964),Microbial Ecology of Activated
Sludge.I. Dominant Bacteria,Appl. Microbiol., 12,412.
Dias, F.F.,Dondero,N.C., and Finstein, M.S. (1968), Attached
Growth of Sphaerotilus and Mixed Populations in a
Continuous-F1ow Apparatus,Appl. Microbiol., 16, 119l.
R.L and Vesilind P.A. (1969), The SludgeVolume Index:
Dick,
What is lt? J. Water Pollut. Control Fed., 41, 1285.
Dold, P.L. and Marais, G.v.R. (1986), Evaluationof the General
Activated Sludge Model Proposedby the IAWPRC Task
Group,Water Sci.Technol.,18:6, 63-89.
Dommergues,Y.R., Belser, L.W., and Schmidt, E.L. (1978),
Limiting factors for Microbial Growth and Activity in
Soil, Adv. Miuobial Ecol.,2,49.
Donaldson,W. (1932), SomeNotes on the Operationof Sewage
TreatmentWorks, SewageWorksJ., 4,48.
Dondero, N.C. (1961), Sphaerotilus'.its Nature and Economic
Significance,Adv. Appl. Microbiol., 3,77.
Dondero,N.C., Phillips, R.A., and Heukelekian,H. (1961), Iso-
lation and Preservation of Cultures of Sphaerotilus,
Appl. Microbiol., 9, 219.
Doohan, D. (1975), Rotifera, in Ecological Aspects of Used-
Water Treatment: The Organisms and Their Ecology,
Curds, C.R. and Hawkes, H.A., Eds., Academic Press,
New York, chap.7.
Eberhard,W.D. and Nesbitt,J.B. (1968),ChemicalPrecipitation
of Phosphorusin a High RateActivatedSludge System,
J. Water Pollut. Contol Fed.. 40. 1239.
Eikelboom, D.H. (1975), FilamentousOrganismsObservedin
Bulking Activated Sludge,Water Res.,9,365.
Eikelboom, D.H. (1977), Identificationof FilamentousOrgan-
ismsin Bulking ActivatedSludge,Progr.WaterTechnol.,
8,1 53 .
Eikelboom, D.H. (1982a),Biosorption and Preventionof Bulk-
ing Sludgeby Meansof a High Floc Loading, in Bulking
of Activated Sludge: Preventative and Remedial Meth'
ods, Chambers,B. and Tomlinson,E.J.,Eds., Ellis Hor-
wood, Chichester,chap. 6.
Eikelboom, D.H. (1982b), Biological Characteristicsof Oxida-
tion Ditch Sludge, in Oxidation Ditch Technology,CEP
ConsultantsLtd., Edinburgh.
P roblem s
Eikelboom, D.H. (2000), Process Control of Activated Sludge
Plants by Microscopic Investigation. IWA Publishing,
London.
Eikelboom, D.H. and Geurkink, B. (2002), Filamentous Micro-
organisms Observed in Industrial Activated Sludge
Plants, Water Sci. Technol., 46:l-2, 535.
Eikelboom,D.H. and Grovenstein,J. (1998),Control of Bulking
in a Full Scale Plant by Addition of Talc (PE84l8),
Water Sci.Technol..37:4-5.297.
Eikelboom, D.H. and van Buijsen H.J.J. (1981), Microscopic
Sludge Investigation Manual, TNO Research Institute
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Eikelboom, D.H., Andreadakis,A., and Andreasen,K. (1998),
Surveyof FilamentousPopulationsin Nutrient Removal
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3'7:4-5.281.
Ekama, G.A. and Marais, G.v.R. (1985), Exploratory Study on
Activated Sludge Bulking and Foaming Problems in
South Africa (1983-1984), Report 54, University of
Cape Town, SouthAfrica.
Ekama, G.A. and Marais, G.v.R. (1986),The Implication of the
IAWPRC Hydrolysis Hypothesison Low F/M Bulking,
Water Sci.Technol.,l8:11, 19.
Ekama, G.A., Barnard,J.L., Giinthert, F.W., Krebs, P., McCor-
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SecondarySettling Tanks: Theory, Modelling, Design
and Operation,Scientificand TechicalReport 6. IAWQ,
. London.
Ekster,A. and Jenkins,D. (1999), Nocardioform Foam Control
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Conl, New Orleans,LA, on CD-ROM.
Fainsod,A., Pagilla,K., Pitt, P.A.,and Mamais,D. (1999),The
Effect of Anaerobic Selectors on Nocardioform Organ-
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71,rt5.
Farquhar,G.J. and Boyle, W.C. (19'7la),Identificationof Fila-
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Pollut. Control Fed.. 43.604.
Farquhar,G.J. and Boyle, W.C. (1971b),Occurrenceof Filamen-
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Farquhar,G.J. and Boyle, W.C. (1972), Control of Thiothrix in
Activated Sludge, J. Water Pollut. Control Fed., 44, 14.
Fanah, S.R. and Unz, R.F. (1976),Isolationof ExocellularPoly-
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Faust,L. and Wolfe, R.S. (1961),Enrichmentand Cultivation of
Beggiatoaalba, J. Bacteriol., 81, 99.
Finger, R.E. (1973), Solids Control in Activated Sludge Plants
with Alum, J. Water Pollut. Control Fed., 45, 1654.
Finstein, M.S. and Heukelekian,H. (19'14),Gross Dimensions
of Activated Sludge Flocs with Referenceto Bulking,
J. Water Pollut. Control Fed., 39, 33.
FMC Corporation(1973), Bulking Control with Hydrogen Per-
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FMC Corporation(1976), SludgeBulking Cure: Hydrogen Per-
oxide.Tech. Data Pollut. Control, Release95.
ActivatedSludgeFoamingand Control 151

6
5 Average for period

^
F
3
U
2 A
I
0
250

ai 200

t 150

E 100
a

50

Primary effluent
1st stagerelease
S 20
oo Secondary effluent

E ro
a

E ,.G1
c/) ''_r. I
/lI
0
7 T
6 I -_-...
U) t) I
Average for period
--
= cn ll
6>
obo 5 ll
/
E fl
xe
z
3
2

0
I
l*u.*-
Apr Dec Aug Apr Dec Aug Apr Nov Jul Mar Nov Jul Mar Nov
-8 7 + 8 8 + 89--i- 90 + 91 + 92 -+- 93 -l-94 -1-95 +96 -
MonthandYear

FIGURE 5.25 Resultsof all full scaleanaerobicselectorexperimentsat San Franciscofrom 1987 through 1995.(From Fainsod,A.
et al. (1999), Water Environ. Res.,7l, I15. With permission.)

These results suggest that a properly designed and
sized anaerobic selector activated sludge system exhibit-
ing EBPR in a systemwithout nocardioform recycling can
control nocardioform organisms in an activated sludge
system with foam-trapping features. The association of
EBPR effectivenesswith nocardioform control ability is
illustrated in Figure 5.27; as effluent soluble ortho-P
decreases,so do the activatedsludge nocardioform counts.
5. Classifying Selectors and Selective
Foam Wasting
Reference was made earlier to the analogy that can be
drawn between nocardioform foaming in activated sludge
and the use of hydrophobic collectors for the separation
of minerals from ores by selectiveflotation. This principle
has been used to remove nocardioform organisms from
activated sludge. Pretorious (1987) suggestedthat the
buoyancy of nocardioforms could be used to selectively
float them from settleableactivatedsludge flocs in a "clas-
sifying selector."He suggestedthat if the float from such
a selector were not returned to the aeration basin, nocar-
dioforms would be eliminated.
Pretoriousand Laubscher(1987) investigatedthis prin-
ciple in 200-L mechanicallyaeratedactivatedsludgeplants,
one of which was equipped with a 16.7-L fine bubble flo-
tation cell between the aeration basin and the secondary
clarifier. Foaming in the classifying selector system was
virtually eliminatedafter about 2 d while the control system
continued to foam severely.While the flotation cell had no
152 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

. O . Nocmdiofomorgmismwashout
Region III (Cha,et al., i992)
(EBPR md nirification)
I Pagilla,1994
tr Mamais,1992
8 A P i ttandJenki ns,1987
€ O P i nandJenki ns,1988
F-l
'd. Q Periodl,1994-1995
6 O Period2, 1994-1995
U
V Period3, 1994-1995

Region II
(EBPR; no nitrification)

UI

Region I
(no EBPR; no nitrification)

t2 l6 l8
Temperature,
"C

FIGURE 5.26 MCRT and temperature regions for EBPR, nitrification and nocardiofonn growth in anaerobic selector activated
sludge. Solid symbols represent experiments in which nocardioforms were not detected. Open symbols represent experiments in
which nocardioformswere detected.(From Fainsod,A. et al. (1999), WaterEnviron. Res.,7l, 115. With permission.)

U)
ch

J
F
,i
a
E^
e
g
o

o
2
a

o Effluent soluble orthoP o

o Nocardioform counts
z
0
I 2.5 3.0 3.5 4.0
MCRT,d

FIGURE 5.27 Variation of average nocardioform counts and effluent soluble orthoP concentrations with MCRT for full scale
anaerobicselectorexperimentsat San Francisco.(From Fainsod,A. et al. (1999), WaterEnviron. Res.,71,ll5. With permission.)

long-term effect on the MLSS level, a decrease(from about
4500 to 4O00 mg/L) was noted during the first few days
after the installation of the flotation cell. This observation
suggeststhat a flotation cell applied to a long-standing
severenocardioform foaming problem with much accumu-
lated foam may initially waste out more than the desirable
amount of solids inventory. In such circumstances,it may
well be good practice to vacuum off as much accumulated
foam as possible,then start using the flotation cell.
Selectivefoam wasting was employed successfullyby
Richards et al. (1990) at the Utoy Creek, GA activated
sludge plant. This plant was operated in a sludge reaera-
tion/step-feed mode (Figure 5.28) with RAS only in the
first pass of the aeration basin. Foam was allowed to pass
from the aeration basin into an unusedbasin where it was
concentratedby collapsing. Foam SS concentration was
2 to 37oand dewateredwell using the centrifugesnormally
used for WAS. Using selective foam wasting to control
foam allowed the plant to increaseaerationrate and MCRT
values (from 6 to 20 d) so that complete nitrification was
possible for the first time. Once the MCRT was increased
to >20 d, the severe foaming ceased and foam wasting
was not necessary.From a previous article about this plant
(Sezgin and Karr, 1986), it is suspectedthat an industrial
waste containing a slowly degradable surfactant was
present which was only degraded completely at high
ActivatedSludgeFoamingand Control 153

_,
foam
Influent
FIGURE5.28 Planviewof aeration basinswith selectivefoam
wasting,UtoyCreek,GA. (FromRichards, T. et al. (1990),Res.
J. WaterPollut.ControlFed.,62,914.With permission.)
MCRT. The nocardioform foaming may have been stabi-
lized by the presenceof the surfactantat the lower MCRT.
Parkeret al. (2001, 2003) reportedthe useof selective
foam wasting (classifying selectors) at seven activated
sludgeplants(Table5.3). They emphasizethe importance
of removinga small amountof liquid almostcontinuously
from the top 1 to 3 cm of a mixed liquor or RAS stream.
This layer can be removedthroughdevicessuchas down-
ward opening gates or side weirs on channels. Such
devices are best located where the natural flow tends to
direct foam, where the liquid level does not vary greatly,
and where surface turbulence is not excessive.
One or more removal locations may be required and
foam trapping at locations other than where it is removed
should be avoided. Surface wasted material should be
combined with WAS for treatment and the solids con-
tained in it should be accounted for in sludge wasting
calculations. Any collection sump for removed surface
material should be designed to allow for its complete
removal when emptied. Failure to do this will transfer a
foaming problem from one location to another (the col-
lection sump). If pumping is required to empty the col-
lection sump, the pump should be able to break suction
rather than to allow vortex development that would pre-
vent floating material removal. The pump should be able
to pass liquid containing entrained air. Suitable pumps
include submersiblesolids handling, positive displace-
ment diaphragm, self-priming, and air-lift types. Pump
capacitymust be high enoughto pump down the collection
sump, break suction, and remove floating material at all
but peak flows. Pump control devices must be able to
function in liquids containing entrained air.
Parkeret al. (2001, 2003) showedthat the installation
of a classifying selector on the RAS channel of the
29-MGD (1.3 m3/d) Haskell R. Street plant in El Paso,
TX virtually eliminated activated sludge foaming and
reduced nocardioform organism counts from 1 to 2 x 106
intersections/gVSS to 6 x104 intersections/gVSS.

TABLE5.3
ActivatedSludgePlantswith SurfaceWastingin Operation
MMF" Capacity, Aeration Basin Surface Waste
Plant Location MGD (m3/d) or Channel Type Surface Wasting Location Disposal Location

Sacramento,CA 180( 8. 1) Oxygen/coarse bubble MLb distribution channel, WAS line, then DAFd
RAS channel
Appleton, WI 22 ( 1. 0) Fine bubble/coarsebubble MLb distribution channel Alternative WAS wasting system to
DAF
President Street, 27 ( 1. 2) Fine bubble/Ml channels Aeration basin Combined with WAS and primary
Savannah,GA sludge prior to DAF
SouthCobbCounty,GA 40 (1.8) Fine bubble/coarsebubble AB'effluent collection
channel
Metro plant, 80 (3.6) Fine bubble/coarsebubble RAS re-aeration basin, end Combined with WAS prior to DAF
St. Paul, MN of aeration basin passes
Haskell R. Street plant, 29 ( 1. 3) Fine bubble/coarsebubble RAS channel Combined with WAS prior to DAF
El Paso, TX
Utoy Creek, Atlanta, GA 4s (2.0) Fine bubble/coarsebubble MLb distribution channel Combined with WAS ahead of
thickening centrifuges

u Maximum month flow.

b Mixed liquor.

' Aeration basin.

d Dissolved flotation.

Source:From Parker,D.S. et al. (2003), Water Environ. Res.,75,83. With permission.

154 M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
50-250mm
variableopeningfor
scum (a)
FIGURE 5.29 Nocardioformfoam control devicefor surfacechlorinationat the 23rd Avenuewastewatertreatmentolant in Phoenix.
AZ: (a) side view and (b) top view.
6. Chlorination
RAS chlorination is not very effective for controlling
nocardioforms in activated sludge becausethe nocardio-
form filamentsare largely containedinside the activated
sludge flocs. Thus, they are inaccessibleto the chlorine
unless a high enough dose (actually an overdose) is
applied to break up the flocs. Such dosesare not recom-
mendedbecausethey degradeeffluentquality and dimin-
ish treatmentcapacity.Some benefit from RAS chlorina-
tion in controlling nocardioformscan be achievedin a
system with an aerationbasin that has subsurfacewith-
drawal becausethis generatesfree-floatingnocardioform
filamentsthat are accessibleto the chlorine.
A far more effective use of chlorine in nocardioform
control is to apply a chlorinesolutionas a fine spray(mist)
directly to the aerationbasin surface.This approachhas
been successfulat severalplants in the U.S. including the
23rd Avenue plant in Phoenix; Stamford, CT; Ocean
County,NJ; Trenton,NJ. (Albertson,l99l); and SanJose,
CA (Ekster and Jenkins,1999).At the 23rd Avenueplant
in Phoenix,spray hoods (Figure 5.29) were placedacross
the end of the third passof the four-passaerationbasin.
As the foam enteredthe hood with the mixed liquor flow,
it was sprayed at a rate of approximately 10 L/min with
a chlorinesolution(from a gaschlorinator)containing2 to
3 mg ClrlL. Using a chlorine dose in the range 0.5 to
1.0 mg Clrll- basedon the wastewaterflow, nocardioform
foam was eliminatedon severaloccasionswithin 1 to 2 d
without any effluent quality deteriorationor loss of treat-
ment efficiency.
At the San Joseplant, a fine mist spray of NaOCI (3 g
Clrll-) solution was applied through a shroudedspray bar
to the surfaceof one aeratedcompartmentof a 4-compart-
ments-in-series,step-feedBNR activatedsludge system.
At dosesof 0.5 to 1.0mg Clrll- (basedon wastewaterflow),
this systemcontrolled nocardioform foam without altering
bulk mixed liquor nocardioform organism levels, suggest-
ing that it only kills nocardioforms at the surfacesof the
Pr oblem s
Heavily
chlorinated
water spray
(Clr=2-3 t11-,
Direction of
mixed liquor flow
(b)
basins.This method of nocardioform organism control did
not compromisethe quality of the BNR effluent in any way.
It may be possibleto enhancethe effectivenessof a
chlorine surface spray by installing fine bubble aeration
to enhance the nocardioform foaming just prior to or
within the chlorine surface spray system. When using
surface spray chlorination, it is important to avoid dis-
persal of the sprayed(misted) chlorine solution because
ofits hazardousand corrosivenature.Further,when using
a NaOCI solution spray,flushing with water is necessary,
especiallyprior to shut-down,to preventblockageof solu-
tion linesand sprayswith the sparinglysolubleprecipitates
that can form by interactionof the alkaline NaOCI with
water and the atmosphere.
7. Cationic Polymer Addition
In the discussionof the effect of reactorwithdrawal con-
figuration on nocardioform growth form, it was shown that
in pure cultures of G. amarae, an overflow outlet tended
to produce clumped growth while a subsurfacewithdrawal
configuration tendedto producedispersedfilaments.These
sameeffectscan be observedin activatedsludgecontaining
nocardioforms. Indeed the microscopic observation of
free-floating nocardioform filaments in a mixed liquor
samplecan be used as an indication that the aerationbasin
has a subsurfacewithdrawal and that foam trapping and
an accumulationof foam will occur on the aerationbasin.
Becausethe hydrophobic nocardioform organism
surface causes foaming, the presence of free-floating
nocardioforms will enhance foaming over the levels that
would be produced by nocardioforms buried inside acti-
vated sludge flocs (Narayanan,20O2).A subsurfaceaera-
tion basin outlet producesfree-floating nocardioforms that
foam more efficiently. The foam builds up on the basin
surfacebecauseit is trappedby the subsurfaceoutlet. Shao
etal. (199'7\ found that the addition of small amounts
(0.5 mg/L basedon wastewaterflow) of a cationic poly-
acrylamidepolymer (e.g., Polydyne Inc. Clarifloc-C341,
ActivatedSludgeFoamingand Control 155

100 q
q

ge0 il
a
oRO 80 o

Ato ;
O

!6 0 60
'E
d 1tl
E
b 40 40
=.^
E
8zo 20
8 ro E
o
0 z
Jan Jan Jan Jan Feb Feb
9162330613

Date.1995

FIGURE 5.30 Effect of cationic polymer additionon nocardioformconcentrationand aerationbasinfoam coverage,Terminal Island
Plant, Los Angeles,CA. (From Shao,Y.J. et al. (1997), WaterEnvirctnRes.,69,25. With permission.)

aminomethylatepolyacrylamide, low charge density,

molecular weight 20 to 30 x 106)to the aeration basin c
6
eliminatednocardioformfoam. Figure 5.30 showsthat the '' ,.
reductionof aerationbasinfoam coveragewas not accom-
panied by a proportional decreasein nocardioform organ-
ism counts.Shaoet al. (1997) postulatedthat the polymer EX

flocculates the free-floating nocardioforms back into the oE

activatedsludge flocs, thereby decreasingthe amount of
hydrophobic surfaceavailablefor attachmentto air bub-
bles and thencedecreasingfoam formation. t
By eliminating the nocardioform foam, the polymer
addition reincorporatesthe nocardioform organisms con- 05t015202530354045
Time,d
centrated in the foam back into the mixed liquor. Here
they are lesslikely to causefoaming and are accessibleto FICURE5.31 Effecton aerationbasinfoamcoverage of dosingI
wasting in WAS so that they can be eliminated from the mg/Lcationicpolymerat activated sludgeplantin Frankenmuth, MI
activated sludge at the samerate as other activated sludge
microorganisms. ary through March 1997), a polymer dose of 0.2 mglL
Polymer addition to control nocardioform foam has controlled foaming most of the time but rvas inadequateto
been practiced at several treatment facilities subsequent completely eliminate it. Increasingthe polymer dose to 0.4
to the initial trials at the Terminal Island plant in Los to 0.5 mgll- controlled the foam. During the third test period
Angeles.For example,nocardioformfoam on the aeration (April 1997),foam and scumwerecontrolledrapidly using
basins at the plant in Frankenmuth, MI was reduced from a polymer doseof 0.35 mg/L (Eksterand Jenkins,1999).
'757o surface coverage (2 in. deep) to a l07o coverage One of us (DJ) was told an interestinganecdoteduring
(0.25 in. deep)by the addition of 1 mg/L of polymer for a discussionof the use of polymer for nocardioformfoam
41 d ( F igur e5. 3 1 ). control. On hearingabout this method and our hypothesis
The polymer can be dosed either to an RAS wet well for how it works, a treatment plant operator replied, "So
or to a mixed liquor channel.Polymer dosing periods of 3 that'swhy my foam goes away when I overdosepolymer
to 10 d were usedat SanJoseto control nocardioform foam to my belt pressesl" [Language paraphrasedto remove
on three occasions.During the first test period (July 1996), certainAnglo-Saxonadjectivesand verbs.l This very per-
aerationbasin foam and scum levels in the secondaryclar- ceptive operator figured out that the type of polymer used
ifier feed channelwere noticeablyreducedafter I d of inter- for sludgethickening by gravity belts, belt filter presses,
mittent polymer dosing at 0.5 mg/L. After 2 to 3 d of centrifuges, etc. is the sametype that works for nocardio-
continuouspolymer dosing at 0.5 mg/L, the foam and scum form foam control and that an overdoseof polymer to his
virtually disappeared.During the secondtest period (Janu- belt pressesresultedin a recycle of polymer to the aeration
156 M anua lo n C a u s e sa n d C o n tro lo f Ac tivatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
P r oblem s

70 isms, M. parvicella organismsappearto have hydrophobic

Average MCRT 1996= 5.
Averase 5.1I d l---I9%-l cell walls and they can stabilize brown, high-solids foam
AverageMCRT 1997= 6ib d | ""' l9e7 |
60 on activated sludge. BecauseM. parvicella filaments are
much longer than nocardioform filaments, they can
50
decreaseactivatedsludgesettlingratesby bridging between
tr flocs. The morphology of M. parvicella is such that very
.=-+o
- ^^
u
large populations are needed to cause severe bulking. A
>JU curved and coiled filament usually grows throughout and
wraps around the activated sludge flocs, rather than pro-
20
truding rigidly from the floc surfacesinto the bulk solution.
t0 M. parvicella was first describedby van Veen (1973)
and first isolated by Slijkhuis (1983a and 1983b). Like
nocardioforms,it is strongly Gram-positiveand contains
Nov 16 De c 1 7 Ja n 1 6 Feb 15 Mar 16
Neisser-positivegranules.
DateE

FICURE5.32 Comparison of manualcontrol(1996)andauto- 2. Substrates

maticcontrol(1997)of MCRTat theSanJose/Santa
Claraplant.
(FromEkster,A. andJenkins,D. (1999),Proc.WaterEnviron.
Fed.Conf.WEFTEC'99.With permission.)
basin in the filtrate and that this was what eliminatedthe
aerationbasin foam.
8. Automatic MCRT Control
Recentwork at the San JoseBNR plant indicatesthat on-
line instrumentalmeasurements of activatedsludgeMLSS
and RAS SS togetherwith automaticcontrol of wasting
may be better than manual (laboratory) SS measurements
and manual activated sludge wasting for preventing acti-
vated sludge foaming. Online instrumental MLSS and
RAS SS analyseswith the datausedfor automatedMCRT
control eliminated nocardioform foaming from the San
Jose plant and the need to use surfacemist chlorination
or polymer addition for foaming control.
It is possiblethat MCRT control with automaticsludge
wastingandcontinuousTSS analysesallows for much less
variationin discreteMCRT values.This point is illustrated
by the MCRT datain Figure 5.32. Note how the manually
controlled sludgewasting basedon laboratoryTSS anal-
yses (1996 data) includes many incidents of very high
MCRT. It is possible that such incidents encouragethe
growth of nocardioforms that then are retained in the
activated sludge system by surface foam trapping even
when MCRT is lowered. It was possible to operate in a
nocardioform-free condition at a higher average MCRT
(6.3 d) than with manual activatedsludge wasting based
on laboratoryTSS analyses(5.1 d).
IV. MICROTHRIX PARVICELLA
A. FncronsArrrcrrNc M. penwcruAGRowrH
1.
General
Excessivegrowthof M. parvicellain activatedsludgecan
causebothbulkingandfoaming.Like nocardioform organ-
The original isolate of M. parvicella required long chain
fatty acids(e.g.,oleic acid) or their Tweenestersfor growth.
More recentstudies(Wanner,I 994; Holmstrom et al., I 996;
Tandoiet a1.,1997)show that simple carbon substratessuch
as glucose, acetate,and ethanol can support the growth of
M. parvicella. Because M. parvicella-dominated sludge
formed in a predenitrification activatedsludge system fed
with substratessuch as glucose and ethanol, Wanner and
Grau (1988) suggestedthat it could denitrify and form stor-
age products from simple carbon substrates.
Neilsen et al. (2001) showed that M. parvicella can
take up and store long chain fatty acids such as oleic acid
under anoxic and anaerobicconditions.They suggestthat
a surface-associated lipase breaksdown the oleic acid at
the hydrophobic filament surface and facilitates its trans-
port into the cell and storageas neutrallipids. Neilsenet al.
(2001) postulatedthat theseabilities give M. parvicella a
competitive advantagein anaerobicand anoxic conditions
where hydrolysis rates of particulate organics by the acti-
vated sludge community as a whole are much lower than
under aerobicconditions(Dold and Marais, 1986).Under
aerobic conditions, floc-forming microorganisms can
metabolizelipid substrates suchasoleic acid.Nielsenet al.
(2001) suggestthat this ability allows floc formers to out-
compete M.parvicella in well mixed, completely aerated
activated sludge and in aerobic selector systems.
3. OperatingConditions
M. parvicella is not currently a common dominant fila-
mentous organism in activated sludge in the U.S.
(Table2.1). We have observedit mostly in high MCRT
plants in upper tier states,(Michigan, Wisconsin,Minne-
sota,Wyoming, Montana,and Colorado)especiallyduring
the winter. In many other parts of the world (Northern and
SouthernEurope, South Africa) it is by far the most com-
mon dominant filamentousorganism in activatedsludge
systems.This is likely due to the presenceof a higher
ActivatedSludgeFoamingand Control 157

percentageof high MCRT BNR plants in these areasthan
in the U.S. While the specific causes for excessive
M. parvicella growth in activated sludge are not com-
pletely understood,the four most commonly cited condi-
tions associatedwith its srowth in activated sludqe are:
. High MCRT
. Low DO
. Low temperature
. Presence of anoxic. anaerobic. and intermit-
tently aeratedzonessuchas thoseusedin BNR
plants and present in mechanically aeratedaer-
ation basins
As high MCRT BNR plants become more common
in the U.S., the incidenceof M. parvicella is cerlain to
increase.
Wentzel (1992) reported that at the five-stageBarden-
pho JohannesburgNorthern plant (South Africa), M. par-
vicella was the dominantfilamentousorganismin the win-
ter. In the summer, Type 0092 became dominant.
Knoop and Kunst (1998), in pilot plant and full-scale
experimentson municipal wastewatertreatment plants
containinganoxic and/or anaerobiczones,found that the
best growth of M. parvicella occurredat <12 to 15'C and
with F/M values of (0.1 kg BOD./kg MLSS/d. Under
these conditions, M. parvicella filaments were 200 to
500 pm long and SVI values were as high as 490 ml'lg
(average, 250 mLlg). At >20"C and the same F/M, the
M. parvicella filaments fragmentedinto 30- to 80-pm long
pieces, then gradually disappearedfrom the activated
sludge. The filament fragmentation was accompaniedby
an SVI decreaseto approximately 150 mllg. When the
F/M was increasedto >O.2kg BODr/kg MLSS/d, M. par-
vicella was gradually eliminated from the activatedsludge
even at <12 to l5oC. Before disappearingfrom the acti-
vated sludge, the M. parvicella filaments formed rope-like
bundles that did not causeas much interference with set-
tling and SVI valuesdecreasedto <150 ml-/g.
Mamaiset al. (1998)investigatedthe effectsof reactor
mixing conditions and the presenceof anoxic zones on
bulking due to M. parvicella. Using pilot-scale activated
sludgeoperatedat MCRT = 18 d, F/M = 0.2 kg COD/kg
VSS/d, and temperature= 14 to 20oC on municipal waste-
water, the following conditions were examined:
. Completely aerobic plug flow
. Plug flow predenitrification (an anoxic selector
followed by a plug flow aerobic basin with
internal recycle from the end of the aerobic
basin to the anoxic zone)
. Completely aerobic, completely mixed
. Completelymixed, intermittentlyaerated,nitri-
fi cation-denitrifi cation
All reactors had foam-trapping features in their aera-
tion basinsand secondaryclarifiers.The best settlingwas
obtained from systems with plug flow aerobic basins
(Figure 5.33). The two sludges contained very few
M. parvicella filaments. The activated sludge from the
completely mixed nitrifi cation-denitrifi cation systemwith
CSTR basins showed the poorest settling and M. parvice-
lla was dominant. The completely aerobic, completely
mixed system settled better than the completely mixed,
nitrification-denitrificationsystembut poorerthanthe sys-
tems with plug flow aerationbasins.
Gabb and coworkers (1991, 1997a, 1997b) worked
with continuously fed and intermittently fed laboratory
activated sludge systems with a range of aerobic and

120

x>
s
80 oS -^uA&.. I
og
06
st MAAAAAAdA-
}E tr *as
60 tr ^oo
>
qn' = ot"
o
^- o Aerobic plug-flow
l**
9r tr ^"
FO
trO A ^ o PTeNDN plug-flow
*9) o ,; loPreNr
SE
A I r Aerob
Aerobic completelymixed
A
A
x Intermittent aern, NDN
6
B
AA
A
I mixed
OA
nA

0 50 100 150 200 250 300 350 400 450 500

DSVI, ml-/g

FIGURE 5.33 Effect of unaeratedzones and aeration basin configuration on the settling of activated sludge dominated by
M. parvicella. (From Mamais, D. and Jenkins,D. (1992), Water Sci. Technol.,26:5-6,955. With permission.)
' 158 M anua lo n C a u s e sa n d C o n tro lo f A c ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
anoxic conditions treating settled municipal wastewater.
MCRT valueswere 20 to 30 d, F/M was 0.28 to 0.31 kg
COD/kg VSS/d, and temperature was 20'C. Completely
aerobic systems did not bulk and did not support the
growth of low F/M filamentous organisms (including
M. parvicella) whether or not they had selectors.
Continuously fed, intermittently aerated activated
sludge (simulating an oxidation ditch type of aeration
basin) had high SVIs due to proliferation of Type 0092
and M. parvicella. The installation of an aerobic selector
aheadof the intermittently aeratedbasin did not eliminate
thesefilamentousorganisms.When the aerationbasin was
continuouslyaerated(i.e., the units were completely aer-
obic), bulking due to Type 0092 and M. parvicella was
eliminated.
4. Control
The growth of M. parvicella in activated sludge can be
reducedby:
. ReducingMCRT to as low a value as possible
consistentwith achievingtreatmentobjectives
. Providing plug flow basinsin both anoxic and
aerobiczones
. Avoiding intermittentaerationor zonesof low
DO (<l to 2 mglL) in aerobiczones,especially
at the point where mixed liquor from an anaer-
obic or anoxic zone entersan aerobiczone
. Eliminating foam trapping features
. A full-scale aerobicselectorwas subsequentlyincorporated
Addng a polyaluminum chloride-basedfloccu-
lant
Recent work in The Netherlandsand Japan (Eikel-
boom, 1997;Hwang et al., 1998;Roels et al., 2002) has
shown that M. parvicella bulking and foaming was con-
t r olled by t he a d d i ti o n o f p o l y a l u mi n u m chl ori de
(PAX-14, Kemwater B.V., Rozenburg,The Netherlands)
to the aeration basin. Dose rates for low MCRT activated
sludge systemswere functions of MCRT (60/MCRT =
PAX-14 dose,g as Al/kg MLSS). Using this criterion and
an extended aeration activated sludge system with
MCRT =20 d, HRT = I d, and MLSS = 3000 mg/L would
resultin an aerationbasininfluentPAX- 14 doseof approx-
imately 0.5 mg Al/L. That amount is on the same order
of magnitudethat is effective for controlling nocardioform
foaming with cationic polyelectrolyte.
Roels et al. (2002) report that control is achievedafter
about I to 2 weeksof PAX- 14 dosing.It was also reported
that PAX-14 addition seemsto affect M. parvicella stain-
ing properties and filament morphology, suggesting that
someinhibitory toxic effect in addition to flocculation may
occur. PAX-14 addition appeared to have no effect on
nocardioforms or N. limicola.
Pr oblem s
5. Case Histories
The following two examples illustrate how the presence
of alternating anoxic and aerobic zones in an aeration
basin stimulatesthe growth of M. parvicella andhow their
elimination and the use of an aerobic selector controls
M. parvicella.
a. Upper Occoquan Sewage Authority (UOSA), VA
The UOSA Regional Water Reclamation Plant is a
27-MGD (1.2 m3/s) facility utilizing preliminary treat-
ment, primary treatment, and single sludge nitrification.
Extensivephysical/chemicaltreatmentof the nitrified sec-
ondary effluent is provided to allow discharge of the
treated effluent to a drinking water reservoir. Effluent dis-
charge standardsand plant reliability and redundancy
requirementsare extremely stringent,resulting in a con-
servativedesign for the biological treatmentfacilities with
a relatively long hydraulic residencetime, low organic
loading rate, and low secondaryclarifier overflow rate.
The biological treatment plant was upgraded and
expanded from an existing completely mixed activated
sludge system that historically experiencedseverebulking
problems with SVIs as high as 600 ml-/g that restricted
treatment capacity. Poor settleability resulted in reduced
MLSS concentrations,reducedMCRT, and reducedcapacity
to nitrify. Periodic identification of the filamentousorgan-
isms presentindicated that M. parvicella was typically pre-
dominant, especially during cold weather.A bench-scale
evaluation was conducted of an aerobic selector (DO
>2 mg[-) for correctionof the filamentousbulking problem.
into the design of the expandedactivatedsludge system.
Figure 5.34 is a schematicof the expanded,upgraded
full-scale, system. The selector consists of an aerated
channel(length:widthratio of 5:1) with a hydraulic resi-
dencetime of I I min basedon influent flow. Dye testing
indicated that the hydraulic flow pattern in the selector
could be approximatedas three equal-sizedcompletely
mixed tanks in series.Following the selectorare two sets
of activated sludge aeration basins in series.The first set
comprises newly added diffused air basins required to
Primary effluent
RAS
Mixed 9.1."1o.
liquor
To secondary
clarifiers
Diffused
air
Aeration basin
FIGURE 5.34 Aeration basin and selectorsystem,UOSA, VA.
(From Daigger,G.T. and Nicholson, G.A. (1990), Res.J. Water
Pollut. Control Fed., 62,676. With permission.)
 ActivatedSludgeFoamingand Control
Selector
I
500 V operational
d 7
) +oo
<-----> difficulties
F 3oo
a
200
100
1986 1987 | 988
FIGURE 5.35 Effect of selectoroperationon SVI at UOSA, VA. (From Daigger, G.T. and Nicholson, G.A. (1990), Res.J. Water
Pollut. Control Fed., 62,676. With permission.)
accomplishthe plant expansionand the secondcomprises
the existing completely mixed basins using slow speed,
surface mechanical aerators.The volumes of the new dif-
fused air and existing mechanical aeration basins are
approximatelythe same.
Excellent sludge settling, as indicatedby an average
SVI of 14 mLlg, was obtained with the expandedand
upgraded secondarytreatment system while operating in
excessof designorganic loading and with periodsduring
which hydraulicloadingwasequalto the maximum month
designvalue.Figure 5.35 presents4 y of dataillustrating
the impact of the aerobic selectoron sludge settleability.
Before March 1988,the existingcompletelymixed basins
t_ using surfacemechanicalaerationwere in service.Severe
sludgebulking (and some foaming) problemswere expe-
riencedregularly,and SVIs typically were>150 ml-/g and,
on occasion,as high as 600 ml/g. The predominantfila-
ment was M. parvicella. In March 1988,the aerobicselec-
tor and diffused air aerationbasinswere brought on line
and the existing mechanically aeratedbasins were taken
off line. The aeration basin hydraulic residence time
remained about the same becausethe volumes of the
mechanically aeratedand diffused air aerationbasinswere
about the same. The result of the change in reactor con-
figuration was a rapid decreasein SVI (Figure 5.35) as
M. parvicella was eliminated from the system.The SVIs at
UOSA haveremainedlow since installation of the selector,
except for a brief period in late 1988 when construction-
related problems resultedin unusualoperating conditions.
Operating conditions within the selectorwere charac-
terized in February 1989. Direct measurementsindicated
soluble BOD. removalsof approximately 6OVoand soluble
COD removals of approximately 457o through the selec-
tor. Oxygen requirements exerted in the selector were
determinedby (1) measurementof the oxygen uptake rates
of samples withdrawn from the selector and (2) direct
calculation from measuredair flow rates using measured
DO concentrationsand oxygen transfer efficiencies deter-
mined by using an off-gas analyzer
159
Construction
y' related
I 989 1990
Year
Both techniques produced similar results and indi-
cated an oxygen requirement in the selector of approxi-
mately 0. 1 mg Orlmg soluble BODr removed.The DO
concentrationsin the selectorhave been maintainedcon-
sistently above2 mg/L, suggestingthat DO doesnot limit
the rate of oxidation of carbonaceousmatter. This rela-
tively low oxygen requirement suggeststhat storage,
rather than oxidation, is the predominant fate of removed
soluble organic matter in the selector. The F/M for the
entire.selector was 4.9 kg BODr/kg MLSS/d while the
F/M, for the first selector compartment was 14.8 kg
BOD./kg MLSS/d.
Becausetwo changeswere made when the UOSA
plant was upgraded- installationof the aerobicselector
and addition of a diffused air aeration basin upstream of
the existing mechanically-aerated aerationbasin - it is
not possibleto determinethe relative contributions of each
modification to improving the sludgesettlingcharacteristics
and controllin g M. parvicella. However theseobservations
are consistentwith the findings of Mamais et al. (1998)
and Gabb etal. (1991,1997a,1997b)that M. parvicella
growth is controlled by completely aerobic selectorsand
aerationbasinsbut not by aerationbasinsthat containlow
DO zones.The following case study provides additional
insight.
b. Northside Wastewater Treatment Plant,
Tulsa, OK
This plant provides preliminary treatment, primary treat-
ment, and secondary treatment using the complete-mix,
air-activated sludge processwith mechanical surface aer-
ators. Initially the plant provided 28 MGD (1.2 m3/s)of
capacity using two aeration basins. Nitrification occurred
periodically, but the capability to achieve consistentnitri-
fication was limited by chronic filamentous bulking prob-
lems with SVI values as high as 500 ml-/g for periods of
several consecutive weeks. M. parvicella was often the
predominant fi lamentous microorganism causing bulking
during cold weather.
1 60 M anua lo n C a u s e sa n d C o n tro lo f Ac ti vatedS l udgeB ul ki ng,Foami ng,and Other S ol i dsS eparati on
Primary
effluent
Return
activated
sludge
Diffused air
Aeration basin
FIGURE 5.36 Northside plant aerationbasin with aerobic selector,Tulsa, OK. (From Daigger, G.T. and Nicholson, G.A. (1990),
Res.J. WaterPollut. Control Fed.,62,676. With permission.)
Expansion of the plant required reliable, consistent
year-round nitrification. Two additional aeration basins
with essentiallythe same configurationsas the existing
basins were to be provided. To addressthe persistent
sludge bulking problems, an aerobic selectorwas to be
providedalong with the two new aerationbasins.The new
basins were square and equipped with nine slow speed
mechanical surface aeratorsand influent piping networks
that distributedthe influent throughout the basins.Each
basin had an aerobic selector consisting of a two-pass
channel with a length:width ratio of l7:l that received
primary effluent and RAS (Figure 5.36). The selector
HRT, basedon a design flow of 14 MGD (0.61m3/s)per
basin.was l6 min: the HRT in the aerationbasinwas 5.5 h.
The new aerationbasinswere startedup in January1988
and, after a 6-mo acclimationperiod, the performanceof an
aerationbasin with an aerobicselectorwas comparedto the
performanceof an existing aerationbasin with no selector.
Contrary to the experienceat UOSA, the aerobic selector
system settling characteristicswere no better than those in
the systemwithout an aerobicselectorover an 1l-mo period
when the two systemswere operatedwith the sameprocess
foadings and operating parameters.M. parvicella was the
predominantfi lamentousorganism causingbulking.
In the aerobic selector system, the SVI averaged
152 mLlg.In the UOSA aerobicselectorsystem,the aver-
age SVI was J2 ml/g. The overall F/M in the Northside
selectorwas 3.2 kg BOD./kg MLSS/d- somewhatlower
than the valueof 4.9 kg BODr/kg MLSS, d at UOSA. The
very high length:width ratio of the Northside selector
makes it likely that its initial contact zone F/M is compa-
rable to or greater than that at UOSA. The Northside
selector removed the same fraction of soluble BOD,
(607o)as the UOSA selectoralthoughoxygen uptakeand
airflow rate data suggestedthat more of the soluble BOD,
was oxidized rather than stored.
P roblem s
To
secondary
clarifier
A comparison of UOSA and the Tulsa Northside aer-
obic selectorperformancessuggeststhat an aerobic selec-
tor alone is not capable of controlling sludge bulking
caused by M. parvicella. The configuration of the main
aeration basin also seems to affect the occurrence of
M. parvicella. A main aeration basin with uniform aera-
tion (such as a floor covered with fine bubble diffusers)
seemsio provide greater control of M. parvicella growth
than a basin with point source aeration devices (such as
mechanical surface aerators).
V. ANAEROBIC
DIGESTER
FOAMING
Various aspectsof anaerobic digester foaming causedby
nocardioform organismshave been discussedin this chap-
ter. In summary:
. Anaerobic digester foaming problems due to
nocardioforms and M. parvicella originate in
aerationbasins(where theseorganismsgrow).
The foam-causing microorganisms are intro-
duced into the anaerobic digester in the WAS.
. Foaming due to these filamentous organisms
can occur in anaerobic digestersbefore nui-
sancefoams appear on aeration basins because
the filamentous organism concentrationper unit
volume can be higher in an anaerobic digester
than in the mixed liquor becausethe SS content
of the WAS fed to the digester is higher than
the MLSS, especially since the WAS feed has
usually been thickened.
. Serious operating and equipment problems can
be causedby nocardioform foaming in anaero-
bic digesters.Figure 5.37 shorysthe results of
liquid phase percent total solids and tempera-
ture profiles of the contents of an anaerobic
digester at the San Francisco South East plant.
 ActivatedSludgeFoamingand Control
Cover
6.3 84
5 .5 9l
4.'t 94
5.0 89
20 4.5 93
24
28
32
Totalsolids,7o Temperature,"C
FIGURE 5.37 Nocardioform foam effects on percenttotal solids
and temperature profiles in an anaerobic digester, South East
plant, San Francisco,CA.
Note the large fraction of digestervolume occu-
pied by nocardioform-induced foam, inversion
ofTS concentrations(higher on the top than on
the bottom of the digester), and temperature
decreaseof the liquid in the foam layers.
Nocardioforms and M. parvicella both lose
their Gram-positive staining properties and
their Neisser-positive granules during anaero-
bic digestion, Nocardioform filaments fragment
and the long coiled M. parvicella filaments
break into shorter, thicker filaments. Generally
these changes make it more difficult to micro-
s c opic a l l y re c o g n i z e n o c a rd i o fo rms and
M. parvicella in anaerobic digesters than in
activated sludge. For this reason, Hernandez
etal. (1994) developed an immunofluorescent
antibody staining method for G. amarae that
made it possible to count these organismsin
anaerobically digesting sludge. Using this
method and a dehydrogenaseenzyme method
for viability determination, Hernandezand Jen-
kins (1994) showedthat the viability of G. ama-
rae decreased only very slowly during
anaerobicdigestion (decay rate = 0.04/d). Thus,
recycle streams from digested sludge process-
ing (e.g., centrate, filtrate, and supernatant)
could seed the activated sludge plant with
noc ar d i o fo rms . T h e fo a m i n g p ro p erti es
decreasedeven more slowly than the viability
during anaerobic digestion, suggestingthat the
tendency to foam is retained by nonviable
G. amarae cells or cell walls.
The amount of foam produced by nocardio-
forms in anaerobic digesters is greatly influ-
enc ed b y d i g e s te r m i x i n g m e th o d s and
geometry. The production of CO, and CHo by
161
0.4
o Fine bubble gas mixing
.100i
'- -
!
F1
E
J
Coarse bubble gas mixing
I nl
Mechanicalmixing
246810t214
Time,min.
FIGURE5.38 Foamaccumulation ratein experimentalanaer-
obic digestersas a functionof mixing technique.(From van
Niekeik,A.M. et al. (1987),J. WaterPollut.ControlFed.,60,
100.With permission.)
the digestion processcan produce foam without
mixing. This is most evident immediately after
feeding the digester when gas production usu-
ally increases.Figure 5.38 showsthat fine bub-
ble gas mixing devices(unconfinedgasmixers,
gas lances,etc.) producethe greatestamountof
foam; coarse bubble types of mixers produce
less foam; external mechanical mixers and
pump mixing have the least foam-producing
potential. Becausethey have much smaller top
liquid surfaces, egg-shaped digesters tend to
allow smaller accumulations of foam than
cylindrical digesters.
. The best method for controlling nocardioform
or M. parvicella foaming in anaerobicdigesters
is to eliminate causativefilamentous organisms
from the activated sludge system so that they
never reach the anaerobicdigesters.The use of
mechanical mixing, external pump mixing. or
coarse bubble mixing rather than fine bubble
mixing will reducefoaming potential. Westlund
etal. (1996,1998a,1998b) found that a mixer
placed in the gas space above the digesting
sludge can be used to collapse foam. Mixers
that spray sludge onto the surfaceofthe digest-
ing sludge can help collapse foam.
B iblio g r a p h a
y n d R e fe re n ce s
The historian, essentially,wants more documentsthan he
can really use; the dramatist only wants more liberties
than he can take.
The Aspern Papers, Henry James, 1843-1916
In addition to references cited in text. this section lists
additional literature about the causes and control of acti-
vated sludge solids separation problems.
Activated Sludge (1987), Manual of Practice OM-9, Water Envi-
ronment Federation.Alexandria.VA.
Adamse,A.P. ( 1968), Bulking of Dairy WasteActivated Sludge,
Ware rRes . 2.
. 715.
Albagnac,G. and Morfaux, J.N. (1980), Traitabiliti6 Compar6e
en Aeration Prolong6eet en Contact: Stabilisationdes
Eaux R6siduairesde Brasserie,Trib. Cebedeau,33,63.
Albertson, O.E. (1987), The Control of Bulking Sludges:From
the Early Innovators to Current Practice,J. Water Pollut.
Control Fed.. 59. 172.
Albertson,O.E. (1990),Bulking SludgeControl: Progress,Prac-
tice, and Problems,Water Sci.Technol.,23:4-6,835.
Albertson,O.E. (1991), personalcommunication.
Afbertson,O.E. and Hendricks,P. (1992),Bulking and Foaming
Organism Control at Phoenix, Arizona WWTP, Water
Sci. Technol.,26:34, 461.
Al-Diwamy, L.J. and Cross, T. (1978), Ecological Studies of
Nocardia Foams and Other Actinomycetesin Aquatic
Habitats, in Nocardia and Streptomyce,r;Modarski, M.
et al., Eds., Gustav FischerVerlag, Stuttgart,p. 153.
Allen, L.A. (1944), The Bacteriology of Activated Sludge,
J. Hyg., 43,424.
Amann, R., Snaidr,J., Wagner,M., Ludwig, W, and Schleifer,
K.H. (1996), In Situ Visualization of High Genetic
Diversity in a Natural Microbial Community, J. Bacte-
riol.. 178.3496.
Andreasen,K. and Nielsen,P.H. (1998),In Situ Characterization
of Substrate Uptake by Microthrix parvicella Using
Microautoradiography,Water Sci. Technol.,37:4-5, 19.
Ardern, E. and Lockett, W.T. (1914a),Experimentson the Oxi-
dation of Sewage without the Aid of Filters, J. Soc.
Chem.Industr.,33, 523.
Ardern, E. and Lockett, WT. (1914b),The Oxidation of Sewage
without the Aid of Filters, J. Soc. Chem. Industr., 33,
lt2.
Argaman, Y. and Eckenfelder,W.W., Jr. (1989), The Effect of
Selector Removal on Biomass Composition, Res. J.
Water Pollut. Control Fed.. 61.1731.
Aruga, S., Kamagata,Y., Kohno, T., Hanada,S., Nakamura,K.,
and Kanagawa,T. (2002), Characterization of Filamen-
tous Eikelboom Type 021N Bacteria and Descriptions
of Thiothrix discformis sp. nov. and.Thiothrix flexilis
sp. nov, Intl. J. Syst.Evol. Microbiol.,5, 1309.
AIV Working Group 2.6.1 (1989),Report:Preventionand Con-
trol of Bulking Sludge and Scum, Koru. Abwass., 36,
I 65.
Baines, S., Hawkes, H.A., Hewitt, C.H., and Jenkins, S.H.
(1953), Protozoa as Indicators in Activated Sludge
Treatment, SewageIndustr. Wastes,25, 1023.
Banoub,A. (1982), Reducing Energy Consumption:How Two
Communities Did lt, Water Pollut. Control Fed. Hish-
lights,4, 11.
Barnard,J.L. (1978), Solving Sludge Bulking Problems,W'arer
Pollut. Control, 77, 103.
Barnes,D. and Goronszy,M.C. (1980),ContinuousIntermittent
WastewaterSystemsfor Municipal and IndustrialEfflu-
ents,Publ. Health Eng., 8,20.
Baskin, D.E. and Suidan,M.T. (1985), Unified Analysis of
Thickening,J. Environ. Eng. lll,10.
Becker,J.G. and Shaw,C.G. (1955),Fungi in Domestic Sewage
TreatmentPlants,Appl. Microbiol.,3, 173.
Beebe,R.D. (1983), personalcommuniation.
Beebe,R.D. and Jenkins,D. (1981), Control of Filamentous
Bulking at the San Jose/SantaClara Water Pollution
Control Plant, presentedat 53rd Annual Conferenceof
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 lndex 183

bacterial colonies, 15-16 sampling,l0

cell dispersion, 16 "TheSandsof Dee,"131
characteristics, 15 secondary clarifierand, 132
filamentous organism effects, 17 suspended solidsconcentrations, 135
macroorganisms, 15 suspended solidsinventory 132
m i c r o g r a p h s, 3 .2 0 ,2 l surfactants and,64, 131, 135, 136-137,152-153
nonbiological particles, 15 temperature and,73
size, 15 tests,50, 54,139-140
nonbiological components, I trappingandrecycling,137-139,Seealso Foamtrapping
open structure, 17 types,131
p i n f l o c , 2 , 4 , 6 ,6 3 , 1 2 8 Foamingmanagement
poor diversity, 22 anaerobicdigesters,161
selector flocs, 66{7 classifyingselectors,l5 1-152
shear conditions and, 3 continuoussurfacewasting,153
solids separation problems, 3--7, See also specific problems flotationcel1,151-152
dispersed growth, 3 mixingregime,161
filamentous bulking, ut-5 nocardioforms,144-156,Seealso Nocardioformcontrol
foam and scum, 5-7 selectivefoam wasting,152-153
pin floc, 4
sludgewastingadjustment,141
viscous bulking, 3-4 Foamingtests,50, 54,139-140
Floc Characterization Worksheet. 25-26
Foamtrapping,137-139,145
Flocculants,ST,88, 129, 158
anaerobicselectorand, 148
Flocculated suspendedsolids (FSS), 5L-52, 55, 128
anoxicselectorand,147
Flocculation, See also Floc
filamentousorganismgrowthand,72, 73
clarifier performance and, 128-129
Microthrixpamicella growthand, 158
dispersed microorganism removal, 62
Food-to-microorganism ratio (F/fvI),126,Seealso Nutrientdeficiency
Microthrix parvicella growth and, 158
dispersedgrowth,62
protozoa, 46
DO level and,73
Flocculation zone, 128-129
filamentousorganismgrowthand,71
Flotation cell, classifying selector, 15l-152
foamingcontrol,141
Fluorescent in situ hybidization (FISH), 44, 47
indi'catororganisms, 49
F lM, See Foodto-microorganism ratio
low dissolvedoxygenbulkingand, 108,109
Foaming, 131-161, See also Nocardioform foaming
Microthrixparvicella growthand, 157
activated sludge, 132
aeration basin accumulation, 132
samplingissues,I I
aeration basin gas phase and, 137
selectordesignand,118, I 19
anaerobic digesters, 132-133, 137, 160-16l
FountainValley,CA, 109
bacterial steady-statekinetic data (table), 140
Free-swimming ciliates,47, 48, 49
causes.2. 131 Frozenfoam, 132
control, See Foaming management Fungi
facility start-up, 124, l3I internationalcomparisonfor sludgesettlingproblems,61
factors affecting nocardioform growth, 140-14 1 microscopicidentification,43-44,45
filamentous organisms and,6, See also Filamentous organisms: pH and,73
Nocardioform foaming upsffeamseedingsources,73
abundancecomparisons (table), 60
s u r v e y s , 5 9 ,1 3 2
t- G
floating substrates,141
floc-bound vs. free-fl oating nocardioforms, I 39
Gasterotrichs,48, 51
floc structure and, 5-7
flotation process, 136
G bacteria. 65-66.111
foaming organismsurveys,59, 132
Geneprobes,1,44
freezing,132 Glycogenfermentation,111, 114
gas-solid aggregates,5-6 Gonidia,22-23,68,69
international comparison of dominant fi lamentousorganisms (table), micrograph,35
6l Gordonaamarae,116
MCRT and. l4l-144.156 anaerobicdigestionand, 161
M ic rothrix parvic ella and, 136, 156-160 foaming,132,133
nocardioform seeding, 141 pureculturechemostat , 154
138-139,140,144,147
experiments,
nutrient limiting and, 103, 131 steady-state kineticdata(table),140
particlesand, 131, 135-136 Gramstaining,12, 13
pH and, 143-144 filamentousorganismcharacteristics, 2 l-22
photos, 134 micrographs,33
physicaVchemical factors, 135 procedure,15
proposed mechanisms, 135-140 variable-staining organisms,23
pure culture chemostat experiments, 138-139, 140, 144, 147, 154 G r e a s e , 6 1 , 6 12 4, 1
184 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

H Magnesium sulfate (MgSOa), 130

Mean cell residence time (MCRT)
Haft,62 automated control, 156
Haliscomenobacter hydrossis, 7 , 27, 38 ciliates as indicators. 48
aeration basin configurations and, 71 dispersed growth, 62
bulking control case histories, 125 dispersed suspendedsolids and, 129,130
dispersed growth, 65 dissolved oxygen concentration and, 108, 109
DO level and, 73 filamentous organism growth and, 7 1, 77-:78, 127
international comparison in bulking and foaming activated sludge, 61 Microthrix parvicella growth and, 157-158
low dissolved oxygen bulking and, 108, 109 nitrification and,144
MCRT and F/1\{ ranges, 71 nocardioform control
micrograph, 40 aerobic selector, 145
nutrient deficiency and, 68, 69, 103 anaerobic selectors, 148-15 1
organic substrates,69 automatic MCRT control and, 156
selector effectiveness, I 18 conditions of selector ineffectiveness. 126-127
staining reactions, 22 nocardioform growth and foam formation, l4I-145
taxonomy,46 sampling issues, I I
wastewater feeding regime and, 72 selective foam wasting, 152-153
Halistilus natans, I7 temperature effects, 117
Hamilton. OH. l2Vl22 Microbial adhesion, 2
Heavy metal toxicity, 65 Micronutrients, 107-108
Hockey puck cell shape, 19 Microscopes,12-13
Hydraulic residence time (HRT), 7, 93 Microscopic examination, 9
Hydraulic short circuiting effects, 7 algae, 68
Hydrogen sulfide, 60, 70 applications and results, 57-75
Hyperion featment plant, 124-125 bacterial motility, l6
Hyphomicrobium sp.,67 chlorination effects. 94. 100
"do's and don'ts." l4
equipment,12-13
I exocellular material, 67-68
filament counting,9-12,57-58, Seealso Filament counting methods
IgepalC-620,136 filamentous organisms, 17-23, 69, See also Filamentous
Immunofluorescent antibodystaining,161 microorganisms, microscopic examination
India ink reversestaining,l6 floc characteristics, 1zt-17
nutrientdeficiencyanalysis,69, 103 higher invertebrates,48, 51
viscousflocs.4. 5 Flexibacter spp.,67
Initialcontactzones,11l-1 12, ll8, 122 limited diversity, 61-62
Intermittentwastewater feedingregimes,72, 110 Neisser-positive cell clumps, 65-66
selectorsystems,111-127, Seealso Selectorsystems nonmicrobial particles, 60-64
Iron, phosphorus interaction,lM nutrient deficiency, 68, 103
Iron sulfide,60, 63 Spirillum spp.,67
Irregularcell shape,19,31 powdered activated carbon treatment, 61
protozoa and metazoa, 4647
sample preparation, 13-1+
K sampling frequency, 11
samplingpoints, 1G-11
Kinetic data (table), 140
solids recycling, 60
Kinetic selection,115
spirochaetes,67
troubleshooting guide, 14
viscous bulking, 4
L yeast, 66
Lagoons, algae growth in, 68 zoogloeas, 66
Leopoldsdorf sugar mill, Austria, 120 Microthrix parvicella, 32, 37, 39
Lipase, 70 aeration basin configurations and, 72
Lipid uptake and metabolism, T0, 156 aerobic digesters and, 70
Los Angeles, CA, 124-125 aerobic selector and, 158-159
Lowry method (modified), 19 anaerobic digester foaming, 160-161
bulking and, 156
casehistories, 158-160
M chlorination effects, 94
compaction and settling interference, 7
Macrobiotus sp., 48, 5l control, 158
Macronutrients, See Nitrogen; Nutrient deficiency; Phosphorus aerobic selectors, 158-160
Magnesium ion (Mg2+), 63 operating conditions effects on growth, 156-158
dispersed growth and, 3, 129 selector effectiveness, 118, 126, 127
floc formation and, 2 DO level and, 73

b.
 lndex 185

filament counting method, 9, 140 Nitrospora,4T

foaming,5, 131-133, 136, 156-160, 5, See also Nocardioform Nocardia, ^SeeNocardioforms; specifc microorgarusms
foaming Nocardia amarae, See Gordona amarae
temperature effects,T3-74 Nocardia asteroides, 135
foam trapping and,'72,'73 Nocardia brasiliensis, 135
Gram stain, 33 Nocardia caviae, 135
international comparison in bulking and foaming activated sludge, 6 I Nocardia farcinica, 135
lipid uptake and storage, 70, 156 Nocardia pinensis, See Skermania pinensis
low dissolved oxygen bulking and, 108 Nocardioform control, 144, See also Nocardioforms; Selector systems
MCRT and F/I\{ ranges, 71 aerobic selectors,144-146
micrographs, 33,34,44, 133 anaerobic selectors, 147-l5l
Neisserstain. 34 anoxic selectors,146-147
staining reactions, 22 automatic MCRT control, 156
substrates,156 cationic polymer addition, 154-155
taxonomy,46 chlorination' 154
temperature and filamentous organism dominance switching, 73-74 "classifying" selectors, 151-152
wastewater feeding regime and, 72 MCRT and' 145
Mixed liquor suspendedsolids (MLSS) polymer addition, 87
bulking control by reducing concentration, 85-87 selective foam wasting, 152-153
bulking control by reducing inventory,85 Nocardioform foaming, 131-161, See also Foaming
on-line measurementand foam control, 156 aerationbasins, 135-136
Monas, 48 anaerobicdigesters'137,160-161
Monochloramine,gl-92 cell-wall hydrophobicity,135
. Motility (bacterial), 16, 18 control, see Foaming management;Nocardioform control
M y c o b a c t e r i u m , l 35 d iffe r e n tia tio n ,6 - 7
"floating" substrates,141
floc-bound vs. free-floating filaments, 139
N floc structure and' 5-7 5-'7
L '
flotation process, 136
Nals sp.,48 growth-affectingfactors, 140-141
Neisser-positive tetrads, 65-66, 68 MCRT and, 141-145,156
Neisserstaining, 12, 13 miciographs,133
filamentousorganismcharacteristics,2l-22 Microthrix pawicellafoamingvs., 136
micrographs,34 organismsurveys,132
procedure,15 pH and,143-144
N e m a t o d e s , 4 85, l photos,134
Nitrate, 104 pure culture chemostatexperiments,138-139, 140,144,147,154
colorimetric tests, 107 seeding,142
selector oxygen state and, I 14 selective foam wasting, 152-153
selector systems, 116 steady-statekinetic data (table), 140
Nitrification, See also Denitrification testing, 139-140
chlorination effects, 89-90 Nocardioforms, 37, 39
DO and, 109 aerationbasin configurationsand, 7l
filamentous organism identification,44-45 aerobic digesters and, 70
foaming and,144 branching,37
N oversupply, 104 bulking control case histories, 122
-
ozone and, 102 disPersedgrowth, 65
Nitrification-denitrification plant,125-126 factors affecting growth, 140-141
Nitrifying bacteria, 67, See also Nitrification filament abundancein bulking or foaming facilities, 60
in situ identification, 47 filament counting, 9, 1l, 140
micrograph,22, 47 foaming, See Nocardioform foaming
Nitrite, chlorine reaction,92 foam trapping and, 72
Nitrite oxidizers, 44 Gram stain, 33
Nitrobacter 4445 international comparisons, bulking or foaming sludges, 61
Nitrogen MCRT and F/)\{ ranges, 71
availability, 103-104 micrograph,44
deficiency, 69, See also Nutrient deficiency Neisser stain, 34
fertilizer, 104 organic substrates,140
mixed liquor concentrations, 106 pathogens, 135
oversupply,104 prevalenceissues' 137-138
requrrements, 103 selector effectiveness' 118
residual concentrations,106-107 taxonomy,46
supply and demand, 104-105 Nonfilamentous bulking, 4, See also Viscous bulking
volatile matter, 107 Nonyl phenol ethoxylate (NPE)' 64' 131
Nitrosococcus mobilis, 44, 4':. Northside Wastewater Treatment Plant, Tulsa, OK, 159-160
Nitrosomonas, 4445 Nostocoida limicola
 \

186 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

dispersed growth, 65 Orthophosphate, 107, ll4

selector effectiveness, 118 Oscillatoria sp.,45
Nostocoida limicola I, 30, 32, 38 Oval cell shape,19, 31
compaction and settling interference, 7 Oxygen, dissolved, See Dissolved oxygen
international comparison, bulking and foaming sludges, 6l Ozone, 102
micrograph, 42
staining reactions, 22
taxonomy, 46 P
wastewater feeding regime and, 72
Nostocoida limicola ll, 23, 32, 38 P A C T.61. 88
aeration basin configurations and, 71 Paper fibers, 60, 62, 88
bulking control case histories, 120 Paramecium sp.,49
case study, 98 Particles. 60-61.70
compaction and settling interference, 7 floc microscopic examination, 15
Gram stain, 33 foami ngand, 131, 135-136
intemational comparison, bulking and foaming sludges, 61 micrographs, 62-64
MCRT and F/1VIranges, 71 Pathogenic nocardioforms, 135
micrograph,33,34,42 P A X -14, 158
Neisserstain, 34 Peroxide, 100-101
organic substrates,69-70 Petaluma. CA. 102
pulp and paper facility bulking case study, 70-71 pH
staining reactions, 22 adjustment, 130
taxonomy,46 foaming and,,143-144
wastewater feeding regime and, 72 fungal bulking and, 73
Nostocoida limicola III. 33. 39 yeast growth and, 66
compaction and settling interference, 7 Phase contrast microscope, 12-13
international comparison, bulking and foaming sludges, 6l Phase contrast microscopy, See Microscopic examination
micrograph, 42 PHA staining, 13,21,69
nutrient deficiency and, 69, 103 method, 17
organic substrates,69-70 micrograph, 32
pulp and paper facility bulking case study, 70-71 PHB granules, 68
staining reactions, 22 Phoenix, AZ, 125-126, 141
Nostocoida spp., chlorination effects, 94 Phosphorus
Nutrient deficiency, 69, 103-108, Seealso Food-to-microorganism ratio; availability, 103-104
specific nutrients colorimetric analysis, 106
associatedfi lamentous organisms, 103 deficiency, 69, 103, See also Enhanced biological phosphorus
bulking and, 68 removal; Nutrient deficiency
completely aerobic vs. anaerobic-aerobicsystems, 105-106 discharge limits, 105
diagnosis,103 iron interactions, 104
foaming and, 103, 131 mixed liquor concentrations, 106
intermittent-feeding-based selectors, ll l-127, See also Selector removal, Sae Enhanced biological phosphorus removal
systems requirements, 103
macronutrient availability, 103-104 residual concentrations, 106-107
macronutrient requirements, 103 supply and demand, 105-106
management approach, 77 uptake, aerobic vs. anaerobic-aerobicsystems, 105-106
micronutrients,107-108 yeast growth and, 66
microscopic observation, 68 Pine+reelike organism (WLO, Skermania pinensis),3'7, 132, 133
mixed liquor macronutrient concentrations, 106 P i n fl oc, 2,4,6,63,128
Neisser-positivetetrad.65 Plant duct particle,62
phosphorus removal, See Enhanced biological phosphorus removal Plastic manufacturing facility wastewater,WY 98-100
residual macronutrient concentrations, 106-107 Plug flow, 110
satisfying macronutrient demands, 104-105 Microthrix parvicella growth and, 158
scum, 7 Podophyra sp.,49
Nutrient dischargelimits. 105 Poly-p-hydroxylalkanoates (PHAs), 13, See PHA staining
Polyaluminum chloride, 158
Polychaos sp.,48
o Polymer addition
nocardioform control, 154-155
Oakland,CA, 136 RAS chlorination vs., 88
oit,61,64, t4I settling rate management approaches,87-88
Openfloc structure,17 Polyphosphate metabolism, 114-116, See also Enhanced biological
Operculariaspp.,48, 50 phosphorus removal; Phosphorus
OrangeCountySanitationDistrict Plant,CA, 109 Potassium ion (K+), dispersed growth and, 3,63,65, 129
Organicacids,Neisser-positive
tetradand,65 Powdered activated carbon treatment (PACT), 61, 88
Organicloading,SeeFood-to-microorganism ratio Primary clarifiers, 1, 88
Organicsubstrates,
69-70 Processheat losses, 65
 lndex 187

Protein content, 68 capaci|y, T9

test, 13, 19 fl occulation zone, 128-129
Protozoa,454'l foam effects, 132
"bioaugmented" bacteria consumption, 44 mixed liquor flocculation and, 127-128
bioindicator, 48-49 operating diagram, 81-82
biomass flocculation, 46 operating principles, 78
microscopic observation, 15, 447 performance problems, l2'7 -130
taxonomy, 47-48 investigation, 128
toxicity and, 49, 65 problem resolution, 128-1 30
Pseudomonas,floc, I resul ts,128
Pulp and paper wastewater activated sludge systems thickening capacity, 78-83, See a/so Sludge thickening
bulking casehistories,7U71, 109 process operating relationships, 78-79
intentional waste solids wasting, 89 scum baffles, 138
nonmicrobial particles, 60 scum formation, 131
storm flow management, 87
surface scrapers, 132
R thickening failve, 127
Secondary effluent suspendedsolids, See Effluent suspendedsolids
RAS, See Return activated sludge
Selective foam wasting, 152-153
RAS chlorination, 90-91, See also Chlorination
Selector effect, 1
bulking control, 90-98, See also under Chlorination
Selector flocs, 66-67
chemical interactions, 9 1-92
Selector systems, lll-127
polymer addition vs., 88
aeration basin configuration, 116, 124
pumps, 97
aerobic,anoxic, and anaerobic,111-112,114, Seealso Aerobic
systemdesign,93
selectors;Anaerobic selectors;Anoxic selectors
R e c t a n g u l acr e l l s ha p e .1 9 , 3 L

t_
air compressor failure, 122
Recycled digested sludge solids, 60
approach for difficult filaments, 127
Resin beads,61
bulking control. 145
Retum activated sludge (RAS)
case histories, 120-126
chlorination, See RAS chlorination
aerobic selectors,158-159
clarifier thickening capacity and, 80
aerobic and anoxic selectors. 12V123. 125-126
flow rate and SS concentration, 83-85 'anoxic
selectors, 123-126
on-line SS measurementand foam control, 156
conversion to nitrifi cation-denitrifi cation planr, 125- 126
required flow rate calculation, 79
"classifying" selectors,l5 l-153
sampling issues,11
compartmentalization,I I 1-l 12
secondary clarifier operation, TS-79, See also Secondary clarifier
CSTR comparison,112, 1l6
storm flow management, 87
desi gn,l l 8-120
Reverse staining, Sea India ink reverse staining
aerobic selectors,I l8-1 19
Rhodococcusspp., 143
anaerobic basin, 120
Rhodocyclus,65
anaerobic selectors, 120
Rosettes,22-23, 28, 68, 69
anoxic selectors, 1 19-120
micrograph, 35
initial contact zones, 118
Rotifers,48, 49
effectsof, 117-118
micrograph, 50
energymetabolism,I l4-l l5
filamentous organisms weakly affected by, 126*127
s initial contact zones, I 11-l 12, 118,122
initial oxygen and nitrogen conditions, lll-112
Sampling, 10-14 kinetic selection,1l5
common mistakes,12 mechanisms, 112-117
frequency, I 1 nocardioform control, 144-152
locations,l0-11 aerobic,144-146
sample preparation, l3-14 anaerobic, 14'7-151
transport and storage, 1l-i2 anoxic, 146-147
San Jose/SantaClara, CA,96-97 classifying selectors, 151-152
Sausagecell shape,19, 31 si zi ng,116, 118-120
Scrapers,132 substrateuptake and metabolism, 1I t-l 16
Scum, See also Foaming temperature effects, I 17
c a u s e s ,l 3 l Septic wastewater management approach, 77
denitrification, 5-7 Sequencing batch reactors (SBRs), I
floc structure and,5-7 chlorination, 94
nutrient deficiency, 7 filamentous organism growth and, 71-72
sampling, 10 Settling rate management,See Bulking control
Scum baffles, 138 Settling testsor indices, 50, 52-55, 81, 83, See Sludge volume index;
Secondary clarifier, I specific tests or indices
analysis and operation, 83-85 applications, 57-58
blanket rising, 131 chlorination effects monitoring, 94
 \

lBB Manualon Causes

andControlof Activated
SludgeBulking,Foaming,
andOtherSolidsSeparation
Problems

Settling zone, 79-80 low dissolved oxygen bulking and, 108

Sheaths MCRT and F/IvI ranges, 71
individual filamentous organism descriptions, 24-53 micrographs, 27,30,40
microscopic examination, 18-19 nutrient deficiency and, 69, 103
micrograph, 30 organic substrates,69
staining, 13, 17 selector effectiveness(table), 118
peroxide effects, 101 sheaths,30
Slime formation, causes and effects (table), 2 taxonomy, 46
Sludge bulking, See Bulking upstream seeding sources,73
Sludge lagoons, algae growth in, 68 wastewater feeding regime and, 72
Sludge rising, 131, See a/so Foaming Spirillum spp., 16, 67
Sludge settling problems, See Bulking; Foaming; Solids separation Spirochaetes, 16, 67
problems Squarecell shape,19, 31
Sludge thickening, 78 Staining, 13, 15,21-22, See also Gram staining; India ink reverse
clarifier failure effects, l2l staining; Neisser staining
RAS flow rate manipulation and, 83-85 anaerobic digestion study, 161
state point analysis, 81 specific filamentous organism descriptions, 24-43
theory 79-83 micrographs,33-34
Sludge volume index (SVI), 50, See also Diluted sludge volume index; sample preparation, 13-14
Stined sludge volume index Starch decomposition, 71
aerobic selector case history, 159 Starch granules, 61, 64
alum addition and, 88 Start-up foaming, 124,131
chlorinationeffects,89, 90,94-100 S test, 21, 70
comparison with other settling indices, 83 Stined sludge volume index (SSVI or SSVI3.5), 50, 81, 83
continuous vs. intermittent wastewater feed, I 10, 157-158 aeration basin configuration and, I 10
filament count and, 57-59 clarifier operating diagram, 82
initial aerobic or anaerobic conditions, 11I method, 52
method, 52 Storm flows, 87
_\
Microthrix pamicella growth and, 157 Stroh Brewing Co., Longview, TX, 97-98
ozone effects, 102 Sugar mill, 120
peroxide effects, 101-102 Sulfides,60
RAS flow rate and SS concentration, 83-85 chlorine interactions, 92
secondary clarifier thickening analysis, 8l-83 oxidation, filamentous organism descriptions, 24-43
selector effects, case histories, 120-126 oxidation test, 13, 16
selectorsystemand, I 14 utilization, 70
toxicants and, 65 Sulfur
viscous bulking and, 4 oxidation test, 16
Sodium hydroxide, 130 granules,21,23,70
Sodium hypochlorite, 89, See also Chlorination micrographs, 32,40,41
Sodium ion (Na+), dispersedgrowth and, 3,63,129 S test, 21, 70
Solids recycling, 60 Surfactants
Solids separationproblems, 1, See also specific problems deflocculation and dispersed growth, 3
application of microscopic observations,57-75 floc dispersion and,63-64
bulking, Saa Bulking foaming and,64, l3l, 135-137, 152-153
causesand effects (table), 2 Suspendedsolids
control, See Bulking control; Foaming management analytical methods, 128
dispersed growth (table), 2, See also Dispersed growth automated measurementand foam management, 156
filamentous organism levels and, 57 chlorination effects, 100
floc and, l-'l , See also Floc dispersed (DSS), 5 I, 55, 128-129
foaming, See Foaming effluent (ESS), 52
general correction approach, 77-78 clarification problems and,,127-130
microbiological vs. process-relatedproblems, 7,51-52 microbial vs. process-relatedeffects, 7
microscopic methods for diagnosis, 57-75, See Microscopic flocculated (FSS), 51-52, 55, 128
examination foam effects, 132
secondary clarifier performance problems, 127-130 loading rate manipulations for bulking control, 85
Sphaerotilusnatans, 3,24,38 mass balance,79
aeration basin configurations and,7l mixed liquor (MLSS)
anaerobic selector, 147 bulking control by reducing concentration, 85-87
branching,l7,27 bulking control by reducing inventory, 85
bulking control case histories, 125 on-line measurementand foam control, 156
compaction and settling interference, 7 nocardioform foaming and, 137
DO level and,73 secondary clarifier performance problems, 80-82,127-130
dominance conditions (table),14 volatile (VSS), micronutrient concentrations, 107
international comparison, bulking and foaming sludges, 61 SVI, See Sludge volume index
 I ndex 189

t_ Tardigrades, 48, 5l
Taxonomic classification, 46-48
Temperature effects
dispersed growth, 65
T

fi lamentous organism growrh, 73-:74

Turbulence, effluent SS concentrations and, 7
23rd Avenue plant, Phoenix, AZ, 125-126, 141
Type 0041, 35,37,39
aeration basin configurations and, 72
aerobic digesters and, 70
bulking control case histories, 120,122
filament abundancein bulking or foaming facilities, 60
minimum aerobic MCRT, l17 floc structure, 7
selector systems and, I 17 foaming, 132
Tessaracoccus,65 intemational comparison, bulking and foaming sludges, 6l
Testate amoebae,47 MCRT and F/IVI ranges, 71
Thickening, See Sludge thickening micrographs, 30, 33, 34, 43
Thiotrix l,27-28,38 nutrient deficiency and, 68
compaction and settling interference, 7 selector effectiveness, 118, 126
chlorine effect,94 sheath, 30
gonidia, 35 slowly biodegradable substrates,70
micrograph, 41 staining reactions, 22
rosettes, 28 Gram stain, 33
staining reactions, 22 Neisser stain, 34
sulfur granules, 4l variability, 23
taxonomy, 46 taxonomy, 46
Thiotrix Il,28-29,38 wastewater feeding regime and, 72
compaction and settling interference, 7 'Ilpe0092,32,35,39
gonidia, 35
aeration
basinconfigurations
and,72
micrograph,35, 41
aerobicdigestersand,70
staining reactions, 22
floc structure,7
sulfur granules,4l
foaming,132
taxonomy, 46
internationalcomparison,bulkingandfoamingsludges,61
Thitorix spp.
MCRT andF/IVIranges,71
aeration basin configurations and, 71
micrographs, 34,42
anaerobic selector, 147
Neisserstain,34
L branching, 17
bulking control case histories, 125
chlorination effects. 94. 95
selectoreffectiveness,
slowly biodegradable
1I 8, 126-127
substrates,70
stainingreactions,22
filament abundancein bulking or foaming facilities,60
taxonomy,46
gonidia or rosettes, 68, 69
temperature andfi lamentousorganismdominanceswitching,
international comparison, bulking and foaming sludges, 6l
tJ-t+
MCRT and F/1\4ranges, 71
wastewater feedingregimeand,72
micrographs,32,41
'I\pe 0211,43,44
motility, 18
Tlpe 021N,27, 38
nutrient deficiency and, 69, 103
abundance in bulkingor foamingfacilities,60

t_-
organic substrates,69-70
pulp and paper facility bulking case study, 7G-71
aerationbasinconfigurations and,71
rosettes or gonidia, 22-23
bulkingcontrolcasehistories,120,125
casestudy,98

t-
selector effectiveness, I 18
sulfide utilization, 70
chlorinationeffects,94
sulfur granules,21, 32 floc structure,7
upstream seeding sources,73 internationalcomparison, bulking andfoamingsludges,6l
wastewater feeding regime and, 72 macronutrient demands,l0,t-105
Tokophyra sp.,49 MCRT andF/]\,Iranges,7l
Total extended filament length (TEFL), 57 micrographs, 30, 32, 34, 35, 40
Total suspendedsolids (TSS), See also Suspendedsolids motility, 18
automated measurementand foam manasement. 156 nocardioformgrowthcomparison,140
chlorination effects, 100 nutrientdeficiencyand,69, 103
foam effects, 132 organicsubstrates, 69-70
nocardioform foaming and, 137 rosettesor gonidia,22-23, 35, 68, 69
Toxicity indicators, protozoa, 49 selector system,113,118
Toxic substances,dispersed growth and,65, 129 sheaths,30
Tri-city, oR, 123-124 stainingreactions,22
Tricklingfilter/solids (TF/SC),
contact 62 Neisserstain,34
True branching, 17,27 steady-state kineticdata(table),140
Tulsa. OK. 159-160 sulfideutilization,70
Turbidity sulfurgranules, 21,23,32,40
chlorination effects, 100 taxonomy,46
dispersed growth, 16 wastewater feedingregimeand,72
pin floc, 4 Type04ll,34,39
190 Manualon Causesand Control of ActivatedSludgeBulking,Foaming,and Other SolidsSeparationProblems

MCRT andF/IVIranges,7l taxonorny,46

micrograph,42 upsEqrmseedingsources,73
organicsubstrates,70 wastewaterfeedingregime and, 72
Tlpe 0581,35,39 "l\pe 1702,23
floc structure,7 casestudy,98
internationalcomparison,bulking and foaming sludges,6l Typ€1851,37, 39
MCRT andF/I\tlranges,71 aerationbasin configurationsand, 7l
micrograph,42 bulking control casehistories, 120
taxonomy,46 floc structure,7
\\pe 0675,37, 39 Gram stain, 33
aerationbasinconfigurationsand, 72 internationalcomparison,bulking and foaming sludges,61
aerobicdigestersand,70 MCRT andF/M ranges,7l
bulkingcontrolcasehistories,120,122 micrograph,33, 43
floc structure,7 organicsubstrates,69-70
foaming,132 selectoreffectiveness,I 18
internationalcomparison,bulking and foaming sludges,61 stainingreactions,22
MCRT andF/M ranges,71 taxonomy,46
micrograph,43 wastewaterfeedingregime and, 72
*1
nutrientdeficiencyand,68 T)pe 1852,23
pulp and paperfacility bulking casestudy,70-71 Type1863,37
selectoreffectiveness, 118,126 dispersedgrowth, 65
slowly biodegradable substrates,70 foaming,5, 7, l3l, Seealso Nocardioformfoaming
stainingreacdons,22 MCRT and Fn\,i ranges,71 !
taxonomy,46 micrograph,44
wasrcwabrfeedingregime and, 72 taxonomy,46
Tlpe 0803,37, 39
I
floc structure.7
U I
intemationalcomparison, bulkingand foamingsludges,6l
MCRT andF/lvI ranges,7l
micrograph,43 Ultravioletdisinfection,89
UpperOccoquanSewageAuthority (UOSA),VA, 145, 158-159
taxonomy,46
Tlpe 0914,29-30, 38 Urea. 104
complementary bacteria,30 Uronemasp,,68
dispersedgrowth, 65
floc structure,7
V
foaming,132
internationalcomparison, bulkingandfoamingsludges,61 Victor Valley,CA, 136
MCRT andF/1\tlranges,71 Viscousbulking
micrographs, 32, 41 chlorinationeffects,94
organicsubstrates,70 floc structureand, 3-4
pulp and paperfacility bulking casestudy,7G7l micrograph,5
stainingreactions,22 nutrientdeficiencyand,68
sulfideutilization,70 Volatilesuspended solids(VSS),micronutrientconcentrations,
107
sulfur granules,21, 23, 32, 4l Vorticellaspp.,48, 50
taxonomy,46
thermotolerance,74
Tlpe 0961,34-35,39 w {
floc structure,7
intemationalcomparison,bulkingandfoamingsludges,61 Wet weatherflows,87
micrograph,42 Woonsocket,RI, 109-1l0
taxonomy,46 Worms,48,5l
T}pe 1701,24-25,38
aerationbasin configurationsand, 7l
anaerobicselector.147
Y
branching,17
Yeast,66, 67
bulkingcontrolcasehistories,122,125
casestudy,98
DO level and.73 z
filament abundancein bulking or foarning facilities, 60
floc structure,7 Zone settling, 79-80
intemationalcomparison,bulking and foaming sludges,61 Zoogloealbulking, SeeViscousbulking
low dissolvedoxygenbulkingand, 108, 109 hogloea ramigera,I, ll3, 115, 140, 144
MCRT and F/lvI ranges,7l Zoogloeas, 1,66
micrograph,40 aerobicselectorsand, 112
organic substrates,69-70 micrographs, 2l
selectoreffectiveness. I 18 "selectoreffect" and, I
EilVIRONMENTAL ANDTECHNOLOGY
SCIENCE

MANUAIonthe
CAUSES,nd
CONTROL
OfACTI\ATED
SLUDGE
BULKING'FOAMING,*d
OTHER SOLIDS
SEPARATIONPROBTEMS
G.Richud GlenT.Daigger
JenkinsMichael
David

The most common activated sludge operating problems causing poor plant performance are related to solids
separation.Especiallycommon are bulking and foaming.'Without a proper scientific foundation to support the
efforts of wasrewatertreatment plant management,many attempts to thwart bulking and foaming have failed.

Manual on the Causes and Control of Activated Sludge Bulking, Foaming, and Other Solids Separation
Problems provides the critical scientific and practical underpinnings needed to understand and combat these
problems. The third edition of this flagship text is a comprehensive,conciseguide to the microbiological and
technical aspectsof controlling all rypesof solid separationproblems.

The scientific theory is applied to real-world scenarios,greatly increasing the number of real-world examples
of successfulcontrol methods. New information is also included on filamentous organism growthand its
application in the control of sludge bulking and foaming. Now plant operators, regulators and wastewater
engineershave a complete guide for battling theseformidable design and operating problems.

What's New in the Third Edition:

. Discussionof the mechanismsof bioflocculation involved in the formation of activatedsludge flocs
. Many new casehistories of the practical diagnosisand resolution of activated sludge solids separationproblems
. Reexaminationof the filament backbone concept of floc structure, with additional focus on strong flocs that
are devoid of filamentousorganisms
. Analysis of bulking and foaming causedby nocardioform organisms and Microthrix paruicelk,
with emphasison their origins and control methods '
. New photographs and descriptionsof filamentous
organismsresponsiblefor bulking and foaming
Ll,b' +?
rsB N r-5bb7D- bq?- 5

lliln
. Results of recent studies of activated sludge and
previously unknown microorganisms- and the
potential'use of molecular biJogical tools in
diagnosingbulking and foamingproblems
LeWiS Publishers
www. c r c p r e s s . c o m llilililil