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Color Atlas of Diagnostic Microbiology

Color Atlas of Diagnostic Microbiology

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Published by: enadseyer on Aug 22, 2010
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Luis M.

de la Maza

~ -

Marie T. Pezzh) Ellen Jo Baron

r Atlas of anosttc biology

Color Atlas of


M,·,· bl I

_ Icrobiol logy

Professm; .D.epllr{filCnn o:r PiIIiVfu!oLog.y

DneCL'Oit; DNj;S~!OO of Medljcill Mior():b~oro.gy UlJihre,rs~ty ,@f C.rulifol:"ltlia, Irvine Mecll[;al. Center nl"ang~, Cailror:n[a:

Marie T" Pezzlo" M~A., FfAAMJ

Adlml'ltl;glJra~j ~m Di!rOCno:r. JJ~;PW"'[(]]eil11 of Ptl~holog.y

fIIi.\I\E!:I:'s:ilty or C;rumm"nla. Irvine MJooliml. Cen~lf!:r Or:iifige,. C='ll~rorn:ia,

Ell.en.Jo B€lm~1!, Ph.D.~ 1:.'( AAM)

AdjulIi1ct A800C],~:ce I?ro[~sor of Medlc:il1!e .' -I!'llivers.wt]' ef C~I:~r""nrl:a. Los AIlge'J~s

Cli1ltcilal Associate Professor or MO.lfcular lM:ic["Obtoilogy amill fllUTlUIlJO.~.C!gy Ul!'l!!.wrsUy or Soul~teT]l CalU:onri~

Los .Amgde~, ,Cal[Q[i1:1iH.


:&1. LDo<Jls B~ltJlJ1(JN:: ~MoI1i CtirlSl'Aldl Cl1mllgn ~~re$ New 'rilrt rtilla~lpl'l13 I'arclM'ld IJ;!ryjgll M.;i{lri'il M~:ti~Q Ctw Sing~QfeSr.;lIffi' Jgk,~ TlllIOnlc Wil;iIiJ~defl

,",I i\ 'ri rues M'i lI'I\or W [.',ntmIlIilWl\l"

Vi~ .P,r,,·-jfialr/,P,lhlit1f1er: Von Ladlg .Ediiior.;' Jromhfe:-~ Roche

,Vm·r~m:ellUil B~Ulor: S!l!m:1ra J, f,arker .Prol00t Mr.tnllgtw· Linda iIJJ[{:Kj[lI.e}' PI'i@dr.oc1Jo!~ !dloor: Ricb :B;'J['be<r ~tf!l"er: ~aJb.etb l~t

MI'.I'J'r.~dLlil'jJW S!!lJenri~s~r: lhefEsa Fuchs


1~IKll'rliitii'!:l ~~

Cuprydgb'[ e 19~7 b I\oJ)o !by-1'b.ar IRQok. Inc.

All right.s re.'il9[~d. No, .part of this :public;a,ticm m~! be .r·epr~duced. stlO:rod. in a rebrieval S,fBtem, 'or transmmed, ill m,y [onn Q!i' I;ty alJ,fwea:nlii. electr'Cill1it, mecluotmLireJi. pboroco~, recor,ilJl1g. (!II;' oti:tNW]s-e, w~ulhl]L1twriWm permi~:irol1 ,[if nw' .Il'llIbli:'i!tn~!;"

~fi£5ioI1. ~o,photoc-om' VI. reproduce sQleiy rOlf iliteF.u1 ,ol)i:" P3f:!1onalll:5~ :Is pcrmlttcrll fill Irnl"ari.es orether IlJrers r.eglstered \lilitlb lJhe Co(ljldght CieanLllcu ~lU.c:r" provfded that the base Iee of $4LOO per chapter pilus S, IO pel" pagels .plit'd dll1!1ll[;~1y 'IDme' Copytigbt 'Cb!~e Center, l i Caml:gress Street; SEdan .. MA I] 19 70_Th)s, consent ,doe-II Dol extend to mber k!iJnds oJ cop: Ing. web as c(lIpymg ['Or gene-nal dLffiibuliiol1, for adv.ertiisiog otprlillmot1La~nl 'PW'~t5:, for treating II!'IE\\i' ,coJle:cl:edi woliktl. 'Ilr:fGr ~es~e.

frj)!1~~, ill1, ~h:e UIfl.~~d Slates O'F A:m_ei"l.ca.

Cnm~jt:i:an Billd, U~ho;gr,B:p1byJooIOI":IDm b1/ ms Gnpbics l'l'irltillglbrndkllg b~' 'Wmw'Ol't.h Pre~;. IDt-

MQ~bo,.-Ye81r: Book, me.

1]" "0 WeslliD!e [(ni3U.!ltriai. Dfitf@ St. I.nLllis, ,["'ii~owru (jU46

iLntNilIi3l1:imla'Ji :standard lUook . ~pnlber (HU5i1l-OIJ.21-1

. .

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~ . .

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nfectlous diseases ceased more than 17 u1ll1m(l~ deaths: ilill 1995. almost one thtm of all deaths lh~~ occurred ~Iii'(]~dw~d{~: that ye~.m: The \oVodd He'.altb Otga~ msatlon bCl!1i listed 18 "]]01111 "mfecUou..s diseases ident.ified during the last 22 yean that am G'Ontdb'l1iMng to these st!i1Jt~sUcs:. 'This omlnous rise in 'in:fectious. diseases L~ CiD(::l)llI"lPIS.lIJiICdi by an a,ilrr!l:ost cql,l,;'llly ~Ii~dln:ni.[Jg decrease 10 the numbers ofprograms availeble for tr,a:inin;g £linic;8il1a,bc.maltory SCi.t'H)jtiS~S and mi,eroMolliog]stiS.,

Dh1g.mWiH.cI·.n.icrolhio'[ngy is a.visua] ,Scl,el1!JCt:; i:Jhm~y t~~ [ecng:n~~ dl:ffinllcl;eriSil lcs bot b. nUctoScaipic~dly and maeroseopiea Lly WS ~hehi)l~hnruk of eompetence ~]] the fielidL. A ;C.·ol]!\ile:nLsnt and ind"US]V('l souree of mi:crobj(ll].ogk~;Bi.I. color imagea Is virtllaUy 'r1Jcmexistent. We hmi'e sought ~o remed3f that s~tuatiaJn ll\llth this volume,

The bookcontams 753: full-color Images. encompassing lallimfiIJtt.ory sme~y :plracMc€ls;,. specimen coUecUon. ,@Judi ~r ansport devices, speclmen handling, ,coLony rnorpllilolog~cs. nniicros!Dopric rnmages, bicli:::hemircal:s, and other tests, and slllsoeptilJiility tcsUng. The microbes that cause human dtsease are found among viruses, Fungi,. parasUes,. and baetesta .• and [h~s book tneludes representaure images from each groui]j,Bm.pl:tasi:s: has been pta red 0]]) the p~c~ures and tl~l ]s mllinJ:miilJI, wwtb jlust enough Inlorniatton to place the phctographs tntn contest. The book is meantto he' a eollectlon of some of the most Imporeant 'V.DS:L]aL. Images In human dlagnostic m~crob].(do.gy;

AU instructorsbelptng students: to learnmedlcal mleroblolngy know how dwrn.~ flcult it ,1£: sometimes to IlndtlTh_;at '·tYIlica~!! lmsge toIllustrate a paint-We expect this book~''llirn] provide ~ns~n;u:tor;s easy access to t]],OEl6 images that are fireq twndy used in the teaehi ng .of dliagno~,tl.cmi.cro~iolQgy" T~le 'book ~ho~rnd be useJul ~o medical stu(h~L1.h,t8]k~l1.g their mkl[ohWl.ogy eoueses, pHnW.o~ogy residents aad lnfectlcus d~seases Eeno~l's. p.lt~ol~rnsilt$arud pbyskial1~, and ,of course, lalbotaWty sclentlsts and druaguosUcmlcmh~oLlilgislts, both be ghllliD:g and revtellv]:r]g.

'1\7e' have ,@j~tel1l.:pl:ed ro lnelu de i!'W ~Ms atlas (hose inla!~es, that OOVL'T i[:he most frequently detected pathogens and 1l108~ fn~qll]JeDJ~ly perlormedtests in the clinical '~~bot·~Vory. lIVe reahse that this; Is ,1Il!iTh imffi)08SfbLe ts!~k. because the numbers rof pal:bogJeIU and type3 of tests V~j'y greaU)( from. sit~ to site, If we have failed ito cover a Sl1lbj ect that YUIl.tbink should be mcluded. please feel free to send us y'0ur suggJE,sI:i(JJl1s, or ,ellen better, specimens btFI.d salnp,leiS, to he rOun:s]de.li~ CrJI" the next edit]O[l ,of ~hi~ book Obvi~m~y;,w,e ask a~~ rof our readers, and particularly our critics; to bnng li!1) our attennon .. ~lly errors that tbey m.ny indo .1 n d~I!I~: book. \o\le t hankall of .¥,ou, ~Iladv~nce for YOU[ heliE',

'The light and lillJuff['1eooe:nt mliCrni!l'(}OP~C p~c~1.lt'es were ~3kenwith tal zeiss Un~\rereal :mh:roscope (O:ll"ill Zets-s ~nc"j, West IG~miany) equi,PrnJedi wmth Ze15's and OWymp,us tOiympus OpUcaJ Ci)r~- i Ltd, I Japan.}lefJJses:~ and a Nfu~D(['l FX-3 5 A camera {N~~on, Cc<rp.,j To kyo I' japan), T!:ruel macroscopic ]l11:Biges W'~r.:€i ()lbta~ned using a N:i.lmnHL camerawlth .ai, Mict"o-N]k~or ]; 5 mm ill/3, 0 3, ~e-ns" and! ,3 oOlITl:ntax RTS, camera ivith a Cad Ze~ss S-'P!!a.nar 6 Omnlll, ]'/.208 mens,. Kodachrome 2 5 P[of~ssrnol!'D.a~ film. O~a5tmlil:[l Kodak

f1I D I' j- N" '\.1 k' ..31 V" I! '. rl rile.-..,..' I" '" 'L, ,f.!:l,_ (P" Ph .' ~. ~

', ... .1,)_", ,li\OC:: ,l!flS~l.!I'. '. ,~w ,I, or ') anLII' ewV,La an··, lC,l, oJrv~a I' llJirncuffi'me :ULUJ!5, . ,Ill' L1.J I, ,,I;, ,'CliO I' l~m,

CO",!, Ltd., Ib'!l~yo" ]'a@iBIIII} ,\\!."'fl'fi1: used fo[ most of theimages, The ellectron micfiog:[oii;llpbs were ta~en 'irvith a phU~~s :iE:M,400 electeon mlcroseope {PI1Uips :E1.'ectf1o.nic Instruments Co" Emdhmren~ The N etl1e.t]ands) and mectmn lma,ge ftrnm SO-163 (8astman. Kodak Co.), 'Tl'We In.agmicaU~ll smtedJ M the legends correspcnds to awrummaJJtet]y that of the p1l'1]]]ted 'figure,Ho~'vler, for c;.dc1l1J<!!!!tion~ the measurement p'rovided in 'the; legend should be us-ed"


Spec~a~ thanks for their extra e"l)r~ goto aIm the s~arf' Oir tile' DivwsWOD of 'Me(U~al Microbiology aU U CI 'Merlb;arn Center, \-\le aee ~pecui~.lI,:Y gl:,ate~[ul to, Sandra .A arnaes, J eanne Btandi.rug:. :M.egan C'od.d. N eil :ID)etweirner. Ka~re E,van~;, ~~'aJn€,t S'hlgeii. Radha Srj:k'u:ll':la1ir; Gr,ace 'Ian, and N[~nas Zru:trui:a:m for their belp: tID prepartng the spec~mens and eollectmg the m.!lll&ell~aJ ft>r niclnsicu Wll this hook ..

II: ~:!~=:I ~:j:::r:~~:~1 ~~~:r.smJf oj

fi!e~r romilfdtm:ent to ,paUen:tcarl3' and ~;etlchi1lW nUi!,fe :[/llS ,bo.iJ,~ .possiMe. A,!~d to iFr,ank re~Jo ai~dl james C. taylor jor ;fhe'~r ~rmmd~n:g sJLpport

~~ ~ I '

~ ... ..; - - -

- ~~

~ ,,"- ~ - ~'-. ,- -,-'

- ~- .... -

- - - - - - - - -- - -- - -- - - - - -- - --- - -

;6 S'treptoc(,I'(;cuc.eae, 37'

12 ,A,IJiCI'l)hf~[l PtiTtl10gens Iso.l'aLe!!r l~md/,OI~ M.euJtWed by TiNsue G!JI~:,urf' or Ot:her Spe~:i~a ,lVl~tJU:Mls, 102

14 NJ.ycology,. 11 3

1 i ,b:'I'I'JmUoseml11gy. 194

It] ::li~:,.I:::='~~:~n:':::ti:n~:=~:::eil:Y:~':~~=

saJe~y of personnel l!'!lorkilingwith potellit~jl~ ,~t~o~ens ~n dw[cru ~ab;lratiJt.ies. U k; maflJdatcd by OSHA. {O~cupa'~O]l~rn Safe,t;y~:i]jtllJea]fb Ad. cf1:eatecl19'70)1 ftha~ :g:lID:irle~ bllfSb~ implemented and practi£~d, ned that protec~ive c4odlwng, containmCIl'[ devices and! deoonaammation equipmene and. materials be ,~'W.@Jffi]illhle: ia tih.e~mrk.place, Procednres .[,eg.ru:dwg bLosafety req].lllllirements andpractkes muse be avaUarble and! fau:n]]i.ar~o a[~ laboratnry personnel,

P-·-ote···-' ,c.·I:-"--·-e-·c C·:-:Io'···'~,hi~g··- ,a-· 'n, ,d· Eq. -·-g-ii'-p-m,e·-··-n·t-:

r . .IV - ' .. I!Ii __ !!II!!!, .' . . - 1 I ""! . --- - ..

An tmportant component .of Ilnieersal P[1OC8.uti(Di.IJ::!J and. 'safe work practices is the dfeotive usc of pr,(J&ecutfe ,gJ]()\!e~, eye ,and mOllt~h pI-QtecC~on, and luiboratuty coats, all considered robe Persenal PmteoUV€ Devices .. Protective: glovesmust be wnrn. wlllene\~er there is ,El, potential [Of' direct contact w.H:h lnlectious materlals, DlspolS !lIbLe latex o;r vtny1 .~~OVf;S are recommended and should ]mme(Uate!iy be dlscerded ln a de'S'Jglllatedi wmste container ~ih.en[lenlomd"

1..111 Remove one gI,miEl h!.' lJil!iJlliJlil:g the dlll.I1iJde bCJi~Ro:m [lJ[ l!iI!Elt gJ~~ it'Ow,111'rn tille' I~mlmtlll!i:i. l'li[l.grn~:s \-\!il:h ti~e o~:hei" gimred handl. UWrl:l1l0!B ilihe gllDl,i'C' lliD!l1ldle· !lJIlJtt as lit·~ .rem~1l'l3cl.,. Flo nnt tmu:;h }'<lll[ skinl wUI:i ~:llc (IlUiCi' :~il\ovi;: sru:r":toe',

't ... 2 ~liill tll'iE glove cmrnpll!!li3ly ,uff iltid hO:~[1 tllil:~il ,itontmlf:J:L· rlJ!llM.glulli®.irlJ the gilQ].;Ietllti!tnCi!. D!ii ]'],[II~ illJ'I!Ich tlle n1;liI!CI" m:riI':ace (If either :gkl'lle 'wilhCheIJ1nglo.li\e.i! 1l'lI:ulL


'1:",,3 rnjsil'llg, t~e I!il!l'I!gll1~d hafl!cl_, gI!:' .. :b the t~Meof rt'hc botrom I(ilr tlhe s~ooll,d gL~ ~J!il!d Ifw_lllili,c other dO'lrC' ,off;


1I-:!!!l, Botb ~~ho"Jid! be ;i;rn]ued_l1JJ~e~ dI~diOO m a was~fl' OOJltmner d_e5igJ1~tcd, foil' Qotl~aJlfmn<lt~Gd maJI mal:) "

11",,4 'Puillhe S!'3I;:omll gr.ove ,(li~lill:' '~'hle ,oon~mmnartoo, gI~ that is hdcl,]11 ilthe h,a.nid. '~!m:nln.g jjl: oompletezyiTIsme: cut to c(JII'I!iain the:ftrst ,gLm.re'i[lId ~o <fi'il'oitill il":OiLl!l\a.ll.!Limdil1I,!, :IJllflll1]]~Jl i]<l!!](!lJl~

'I-a ~oo\le]il1g sueh :!Ji.'!gnggles s:!wtllcl' be 'WOO",11 \~Il~ \\'l""Ol;kil1g 'wilhb~y ]lul>il:s, o.r otlbcr :speC1m>CIlS th~t ma'lf Sp~,B.s-nl 01: proced1R'e:;: t:iIloJlJ: 1iIHIJ}lC':refli~ aerosols. Gdlgg~es wrlh :~Jde poo" els slwwd b~pro~ided for 'c;l®cIbWQrker,

11-7" A [19oemnsk ts 'worn to [WOtl!Ct 'm1lOC0'I..'li1 ;mOOllhrD,er.: (If the ni]searlic!m!lJ'I,l!ih ;)ndi toscreen !IYtl:t. aC['i:!wJ lMIil'ticles ~lUlit Ol!~ er:uf~~tie ~~Oty('lit'~~-e~:mv~ UB~~1'ihf: ~lIDa~ should! .fI:~ U~tly arnrJJJrui the face: and ~e~ tib.e OSfl1i."maM .. t~ 1,e~Jl:()f ]Yrm:ect~Q]l."fa,ce ma~ mullld,ru.w-ays be 'I'IID['o,:hr tue-r,e :is >!!l. eh .. n~ ~f sp~U.et'illg d~llicflll .mal!1lrl,al. In ~iOnle seil1il1g:s. waterp.rooffare:shwlds 1'I1,zybe[e~II;I!il",e(I fb-l":.mme tasks gm'{~~!.1- iI:~'g serum ;illIlcl. b100-d.

1-0 Tb.is p:&!)otog:ra~!b. n:raJsl.r'BI~es '~IDJI.DInp:[{jpedy Iii.H~d 1il)(1I: 11D:]I* ~h!li does 1Il000plJO,virl'e :lIdsq;IJj,~te p:mt.elEhIllIL 'ill'.here is apt], t~llal For oolillt,.rnllllatea [n~tcW'''Il~ t() enter ihrlJ!l1gh the g;'l[lf: and ,ooriOe iJ1ID !(:onl:a.ct '\l,Ilth IntlCo:!.'lS m~llibrM1es orto be hIieff[fued i.n..

t"10' A :ITl.:Iill rllUJ!il~hi~'l{i tCSVlfCJl'l:I t(!l'protoc( 'tb.'I3'I~S. nose, ami moulLlm r[l(JI]iJ ~IU5hl31i,

1-11 i?iltild-I:',esistOi!lt I.tborato:rywal\,: 'wlib d(Js~ .medll:s HfLtd '~vrlirt'8 i5~j(i1Jlld be: '!ili'!:llm ttl! IQOl<\rn' all (;I{i'te:i1Ua]i I~d 8kiHlH.ttl!d cw[fu:L~g.Flu~d-:r'~.sta[it liOIbor8[!I)[)' ooaLS siluuM bewern miEI:" :silroeel ,t.+othBtl and sool.ild be remlQVcd beforele''l.viJ'Ilg the' :I aborniiDry..

101l:1!!: 'Knit c:u:1if:! pWI£:Q::U tile skin nn lhe W'lillIS.

1 .... a· 'PJ)~ltl'.Cti!.le gJO'I,/JCS S:11Oli.J~d be ]JIfllJI11 iI.lJi{;"( the' knit C lJiffs to p[,o\fld~ a pmper sleeve clu:rlllli"G and .ltand pmteclion.,


1-1., D 'onurlanull:illJllol'll of wOld:. Sl1!l',F3oes al [hi cna of ea n, ~hlUI and cleLnillg of s,-p,ills should be perlilJfl1led ~\!ILb ~rJreslJJ 1. 0% sott]UlDrl. ofhi)I;Ji!iE!no:t!d hl~h.

'1-'15 Paper W!i!i cl hOl!l.lr:! be. placed 0[1 the oontaminalroo wli:lrk ~1.I.~r ace <llJIti soased w'iiil diJuted hOllSC.'i'm:old hle1:ldl imIlflMiild.ely ~fter 3. spiU md! <It the end df each :illlft.

1"'118, Bl~acb..-ooaked. P\iI[X!r ~,O'iir\lcls :sho.uJ{i tihlo 00 used ln the blo[(pgk<L1 iSafd~r c<ibim~t to' cat,dl, fallcn droOps mal carr creal aerosols ..

W'esb! ID:iSposall

Ca.nt8l!m:i.iIJ.~11Itr:':d materials sb 00 ld be djSPO~6d of i III proper containers that are pro:pm cdy labelli,ed and fltted wlth lids, Separate containers are availaMel:i):r pia stic and glass andneedles, n i~ ~mpnr1:limt ~o dispnse orf m,d1terl!ah, i.n p[101}1~:r rnlnt~jl1ers to avoid potentially serious ifi]uy 'Or wUae:Js ~Q lndtvtduals (Xl]]1m;g in ceatact w:ith these materials.

"·'1117' Ag.m:- plates. af"': diiscarded i't1i. a lI?UQpe.lrlj' laimled CO[l1- ~a[[Ie:1r Vi'it~, a !jd.

'I _'lIS Needles wlI'Dlh attached S.l'.i"lil1.g,es: are discllIJaca into h:ilI'dl">.~rlled,. pmu:lum.proof ,L".Ontai!ITlers_ Nood~.e.'I1mm.ll IUllICr he recapped unless a spcc.tally de.~Jg[led. c(m]I'll~I:J'1 ~ ,;::IV;(I ~I"'ble hoildi;!'r .rnrresn)()QtiTI [I.gl1;~n~ Is: ulSed flj1!'lhatptlwIXI£e.

1~a GlatSSbubli!'S; wllhri.[JJinM, human .sm:nup.i:e;s, araalsn diiS~ carded .illliu banl.-skled, pun.clu[,e--p:roof iPOllhI.Lne.r:,;.


An malter"aJs contaminated with po entiaWly infectious mlcroorgani ms mu be decontaminated before diposal. An autoclave is the most common mS1~\Ullent used to pedorm Uns Iunctton.

11-:i:Q' lJ(UJlSebr pm;;ked] materials Llud, ha~~ bC:Sil. appro.p'ri~ awly dlJsc<1fdcd In ,dool&lll'l,tl!f"tj.I;;QlIwlnJers (i.e" rBd~l:m,gE!l;l~llil:reLi buckets, all' placed illl ehe ,mtodavc). The b<1I;gs,~-\I'hidl1. should neser bcfillled CliJ1ll1:pl-lel},. are loostily He:d at the top to illkmr the steam to c~[iL1!lhJ'le and e:lre~nwjy doo tlJ, rnil1at til 000- teats, Adding appro.Ylmat,ely 1 CLilp orw,~l.er 11.0 Lbi:i ~a,g facl1l'r:a;I~ :'l~ca1ln pm(J)llc1irnl.

-a'l Og".C8· II ·S_.- ..... C' s· b·II.·eI· ,

. ,""., '. I .-Cl!!!'G'., , ..... :,'. l'~.l,

'\11,,211 The iliilloctali"C door Is eli} ed tfighdy.. A~ the oomp1cLi[IJ"i of the ~I@" the dl::H;)r" lis 'Op.eIlOO slowly by iilIJl (lp~rawr 'w.earing heat-resisE<li_fl1 gf(w~.s and a face mask 'lit} prollCCI: agalnst briJrifiS: frUIIIIl, ~<lpmt ste m,

The blologtcal safety cabinel (BSe) is one 'Of the most common eentalnment devlces used In leboratories, Air ].$ deeentamtnated by h1:gh-efLtciency partlculateair (H UJ'A) filters,

,,·,a. A Coollillonllr used CIa s [[ biolOgtcru metJ' ,cabinet tbHt. liILtlr soodlllii.ls both the lriooming andl cxhaosk:d air . .i\ BS ~hm;liill be kEl'pt rUB of r.:lIJ:I;~er, ,[roufule if:lSI;:u::.cUliJu QJ the cabll1ct by a cerUtied 'techoi'CUHIl.is r'6quilrud.

11 .. 23 ll)~Il~W: ml'l~~uj['m;g of aiir .Ilow S.bO'LiM ~·pe_rrOi'ili.'i~ ~CI ensure pOOJ!el" \I~IIOdl(j'1. Mijcll::l!Dk~~. p.rol:ile:lJ!llS, .m.tGr dOQg)Llg" OJjnrl 'Q):tiler [~~ 'COrJitr:ifulf!]te to lonre:red velocit)'; w:li:Jlch m~ aJ~mv lmocUm.lsparHcJes hl esc .. pethe cabiaet,

Use of aerosel-contalnlng safety buckets In eenmfuges, ,?ode::dng tightly sealed tuhes: ina biolo,gil: a] 5afe1tyGiilbrnnd (T3.S!q, aad securlng gas ranks to a ho:lrling device H10U nted an the wallare other ,8idldHio[['i'IIJ safety practiees that are requlredto ensure 1:1. sHfe' worl en1!l'~lro~!1~I]]!t':nt

1 .. ;24 'TJ;gbitly se.Oi!Ioo ceilJtrifl[~~r!!bel>arr'f: placed iI1~D sru-e~y carriees Um:l( iI!re UlJil:m:lillJ!l.lI\IilS lig]1b~Y seiillled !J~~Ul'!.~ cell!~Jij[~~'ttio:n. lhis wm ;:l1,Klid clre@l.i!JJ!;l: a!'l! ;;!.ew$l;d Ui :~LD,~~. are broken

d.!lil'I'lI.l1g centrlru~.t~ol1. ..

1 .. 215, ,Ca!"e sheuldbe tt.M:.em. when. opeOii~1S .!lI sealed Oel!ll!t[iru~ C8lrrier iI:oiWWd eentaminetton (rOll;ll i8.1J'i)sitbte b!,1QI.:i~ cOfi~"'it1!~ dUit'ing oeTlib:i£JUg&~t(!i!'h

1'''~Q Tigh:lTIj! sc,<llcd oorl!tatl'iJCit's sJI'WIJ_~d :be '1\':ClJrtfl!(er!~n a tt~C orujt'll\l[clnbe sbo'llM th_C11 be 1Jj,'llcrl:cd to reabsorb acrow~l!7:ed pa:tttid£'S" A 30·m.iJ~.iI.i~· w\t'i:i~, befQre opCU'lLi1IS '~hernbc ~No~ds OiII~IXIili"u~ wJ<IIlio:[t,!.1spool;rlily H't.~ijJ illJ,<'!JQbm':I~[iiJ aft bein!ll.ltau" dl,edl.

~-27 A~!~ gm'l bilE :should be secllJ.ood. i:n'I P1ill.ce by ~. ilolilillg (I'evtce,

Poli;ential~y inEoct10US material must be shipped according to the federal codes 0[' Interstate SlJip.nl~nt of Btiolc;l;g;ic Agell'ts.

1I .. 2B The spedm~. rn:tl'S"~ be p]1)C'Cd. in an ilRL1~r sea[ed conlalllltlr; w~ilLich tstheu ~Q)la(ledl ill1 an outer stalled ll~etalt:1!l!:be, 'l'he il[mf:lr Cim:al<lIilo.!.rr IHLJI.::.t ~,l'~~ ::mlltieJflt ma.l;c.i"iocl ]0 oIDsotb ithe e.rnl.nm I1uidlool:lt~[n:I;li in lh~ '~n~IJll1Jr <J. teak.

, .. \29' l\i.IJ oWeial ~~i"}(1olii~ A!llJU:S label must be mrL~cd to '[he outer 'C"mT~arille:r,

CflAP'TEB 2 Specimen Collecti,o'n


n tt is the responsibiUty or the dlagnostle mieroblologylaborntory to select and . pm't11de transport de\flCeS ~orco]ledton of specimens rr-Oll1 a varwdy of an ~]t(ll:m:ical sltes, Most s:pecilme]] collectron eontalnersincerpur ate transport media

d:mt support the 'vtabMllity of ml:cmorganis.nf'lS encountered tn d~.r1~!c,a] s:peeib]::u;~~:t"l.~l1e' i.ntetprehd len of iDCSldts ]~, c]tl])C'nid{;!int UPOiI1 the Iq U a [~ty of the CQI1;ecced5pecin1e~ and the transport condUious:. Improperly transpcrted specimens lna~{re-sll1h [[1 £3.00]·' ure to isolete the caJus!:l!i!ivemicmorrgan~gm., One of the moot comrn(H] and wldely used; transport .sYStems is a p~asmic tube containing a sterile p'o,lycst~r-t~;p'pcd swab and :111€!(Uo.m to prre1fi1ent the drJ1~~lg ofmic[o(lrgauiSlUS, matntala tlle!l;lHIi and mlnlmlze aver groWltt b .

;iZ"'I ,::!iterlle, !dmpDBHliila: 6eI"Gb'I:c CUIt:U1F6, 'Cal" rectilan IIIlnd uan~p.Dft: syatiem CO'l1s'iil:~Ul1g of It P'1.~~s. de tube: C(ml;t:i illtng; t'i\!"O ru.YOI].~j!pp~d ~wabs m:u:1I mm:lifkd . . ~m:mr['~'i tr,amlp;ciili"i m,edl.lIm!n 3 SCjlio']{r~te amp01ll Ie fRlllL :M.iCflr 'b~o~:Ogy S¥~~Dt5~ (OCkey~filkMrl,). The ,a~nl{! Js :tlocnl'OO (1.1 thcb!:lse:of tile tl.lbenlll;(li:S:~~b"t:aoy aprotool1w sl~'e 9:nlb~ o!l]Lsid:e. This ttnvPQlII.le mustbe crushed al'OO!,' me s~crme:!1 ls QoUectOOi, and th~ :s·waJ,;!! ~ p~.aced bact ILl1!lo tm oontorlhne-r tiD keep fbie ~pec!me~ Oit] U;II!: :$wahsmonst. 'I'@e use 00 !:.)oXOOI1 :s;!N"ab$ ShViidd. be ,.l1Ifivdoo b~auoo·lhl3i.'J.' {j(lJ.U<i;~in rauy i:IIdd5 Ulill HIm! itm itdi.Dblu,Y l~ ,iilli]jL~bitCli.'rDi.iil.

:2 .. 2 NaSIQp:hiBitynI9IBBlI"uFOgBnJ,tBl swab (11;111 .. ghii'W'B'b 'l'y,pu IV; ep~I:'\~'" DlpgllllQt!;tl~81~ C'18Iil1lWoac!l. lin.). l'l'lliis fleHi~le w[r{!' :l,h.~ft.~lndi sm!ll1lt~.f1lJOC!vldes c1Isy s[J6Climcm lCo!loc-illJDfl. espoci<lilily [or :r.t fJJBOIl:1'~ltJir~ ngeal lllro 1!Ii1~ le urethral S[l}ecimOtlii'l. cruc1um R~gl.MLe swabs :s'Jliuoillld 110:[, be uood fur ,ooU.0o1J:on orr iMethrall: !>pt:ldmeti~. beeause tiu,ey C'iUIl lJill wJ'li't:c ['U' sOme strains tlf Nd&'l<ll'rill OIl'liOf,rJil'£it.ltl:e; Il1m1r-eoof. rhto/ !dJ~ lli~lI:d [Olf' cullel.::ll~D oJ QrlJrl.il':!lPli1{1 t\Md~aJ:rrfl;rj~~ cL'llttlres.

. ' <, '-. "<,

--- .

',~ .. _ ... _.:;;.;..~

2,.31 iNaM'plilarytiJg&al '!C>ulH!etiOilf!! '!!J~M!'!'i. l\ SifW1!M C!l)lled.:iJo~5:'Y.5lte:nJ as de,scrilbed ~IiFi~~ 2·2: h!)f'leUer; 11i.Li> :>'l1l1':"db ffi lI1I.'"tld f(ir ,u[t,,~.i.o:ll 01£ :tHlSiGll~J"YJ]~] s.pecir[li::D~ (Of re~ry of &fdt:lli?!l:1I .p~rr;ms.t;i:l', 1~~~ toJlX: COOl~Amie-s' ehaecoal IT!iI.Jt'j:,I;IlJIrt. ·metiwlm. \,,'l:I!ic~. 'milsl:i!' ,:;i~ppOX1~~ lh~ ... iabilll,Y or N:e.r~mJrkl' go II rirr'll a&Ui',

2-4 sterlne~, dls,PD981b.1'e,collecllQn a:nd '~mn.9Dn :l!Ipiem f'D" INGa,velY ef: viruses, caOL MlcrobilOIQSi" S:ra,tamsJ' GDa'Imy.9fltlUe, M'd~.~!. Thf'! si.\!ah and hcddcr are shown ~e-pa:r<:lt:e~}1 at the top .afnd :as~e~lbilled a'l: Ullie bolmom, The tuhe· C:{illita:lIJ!S a !mll'o:n~ tri~~ed1>~1<!_b 31:"nd viraJ tnmlJpad medium (lOtlJ.'t..'i.[ilIing flruik's:' :baJa.ll.t!:.ti S"'.itl, :s.olli.tl1~j} altd ~~ti;n1ic;robiC'S'.l'l:iJi!!i ·1l[ll;nrenJti!l ~Cii_.. :nr.tB:rndrying" I:wlp:; ,[lJiaimlail1 vir.@!! viab:lilll:y, aUld !."eWird·::: the gmwlh fir ulher mJtwIJi.al lllmtam..itlliillr-.ts •

21..5 H'I!I1iPB1!I call\eGtIDml IHt oordai!lJ1ng, 1'w stses or' ~lOlcr,or:HI[[JF-";l~: 8i~vab6" 3 sli(l~ [OF pl'e]JHgj1~lial]1 aU a droom :'lm~' at rhe '~iltle ofooIlocUon., and .. siMe 'HxattilOle (8yva Mh:;[',oTr,aJk. S B.!JJ .rOSB, c.Ei.lhf.). The :sMe wID:be stamod J::!'l(!I" iilltillc 1:'II"'ioratGI",li' wifrh iii! chroct I1uClre;!;celit ,;iiiJJtilxil.ll ~a.m.

2 .. a CliPmmydis irlsc-bO!lflo!!lU!!!I coUee.t:rD'1i1 11111:

CQII.taiIl!iIL1g l'tll.'u s:izr:l:'l cd ilacriIJil.fitWCd swabs lLnd <lI ,q:I"Obt:'Lllsh, a :!:!jl,"l,e for preparation. of iI. dirooct t!ilW!!tf an:, the 'time of eelleelioii.'l, and an, ~.mpoole Qr ,slli,de ft~ati~ (S~,,fa 1J;oficro'na~. SI.m ffo:!iL't, C1ul!i,), Th.e sli.d~ 'iiN111 be :stoIDlJed ]OJte~· w.lhe l:abo:rarou ~'i'ith a dl:Dl.'.'!Ct [~u:-oOO:'.>(~r.ul, iiDlilibOOjr ~tal[l,

2~7 'VI!rllill."Chlllllllimydl.1 'ilil'ango:t:i; fI'!"!!!Im., ~po::. il[lE:IJIS ror detection O;f'",iT:i\lSes Of chhi~". :sib..ould be placed bn~ ~ tube of s!.!LCm(le"'pM$p~<ne b~~fe-! tr<.lIl~~Oi:lt. 11i1~t1ill.IJti colJta.ming fetal. calf :5"!I!r.um, b'l;~.lIcr,aud ,gliln:m.luciln (.BiilxfQet Ul~osti.cs .. IDe .. [)!i:.·~[,~ikl 111.)1.

a·iQ .I!!iIQ-hg ~vlf'QnmQnt«~ :_lMelitl ~IBbl. MI!:cmbl'glo9¥ Syat'E!m!!J:I' 'CQe'key'a'¥lIm, [Md[.)., nco 8.tJ!~clmc:rn fmr tilla.amb'le: C111tl.rn1: is ('.oUec[e;d as i:isir!il€ i~ iN :stmlJe: lllb~ or, iflliudl.le5.il.ae:s5mb42. om a. Sll.l'l'IlIi. lilfi.d! ooco ]'ih:ioodl in [!TIe t8:s ir.nl~rmeable mvlrurn:m.rm:l;;;bl·ba.g t!l:<'il ccm:l:aiIls arapoulBS or :lrl[BkH~O:F', ctlnllyS'~.al1icl hycllrug:en.C~ g'etrner<rtor: l'h~ billg is Selll;~ed ~:!],[I] each aJlif:lpO'llle j,5 eruslred '~(lo!JrOd[lOe OI_ll1O'!:era.. hlt ,cnnil~tlD[J.<':.



.- ". _ -.....;;;;iii'=-.L


2,,8 At TrF'8m111g ... 0W bufikl ~)BB!L, [!'JII;gJi'ObloIDD &yahl,mll, eOClkJ~ypl]lig~, Mel., for ['IecO\~llry of Nei'~ ~1L1' go'r!(l'.rT~OOil. 'rhe 1.1l![))er HOlt $lU'f;lU.J~ or the botUe' oolltalms IJlfl~li!ie~,,[,,h.~r-M;n:i~Jin ag.-:lI1:' ll~l1Jt !:iJrlill been ~patOO under COlo. 11!!i1i'.J~ft'lbec plaits (BBL Mlcrabla~GlIg¥ &ys,· _ma:ll, CQckieY,li!Ii!vme~ M\CII.) rll't teoo\'€ry ,[:If Ne.i'S.'Il:eritl IiWntii!'.rII~e. :M€d:ulIIffl 1iS .pl1epilred ~.n II. r~!tl. pi.m:.'l:rtc: CO:!ltaimer. ?.lU h iii Sl'Da~-tolll:ii1. lite :lper.i.merD IS i'I1OC'IJI.~ated dlrecl:tv, and 111L'! Il!UJ£sl!:!t:l.te ~ron] the m~rl.i.lJlm disool'lOles a I:Jrc~:rb(l![I3te Utblel li!:J L'tgElOO' ~t.iJe COl: :iI.tlmmsph.er-e~

2,,,',0 p,ar,a .. p'(tR IP1aNlI.ltaIQuY Collectlun Kit (Mgl!:"ldlan DIIIRI1i10.tIC •. , ClftCllilina'IlI!" Oh'IQ)'. This -oom~mlaUypr~rod WI eontatns one l,i'j,ulll)f I!}J:gd:LEiJea poly~liny~ w(lobolIPVA) and one 1<1£1;1 o'f buI'Frn:t>d Dl:lul ml rUII"l~J.aJiln, Placing the Sl(:lO:~ ~pecim.ef] iota these I\riak mWl'ledl!:'Ite~! ,~ur· ~~ 'Q1I~tlm~ preserves ~Jb.t: Il:ltorpholngy or the par:asute:s"

There' are a v,aJ:'"iety of media fmr the collection and reeovery ofmic:roorgan.i.sJ]1s from blood, Some of thesemedia can be used ale ne, while others require i!l1culllatruon and detection tn aD automated 8]i'Sltem .. They aU. ccntaln between 0.015% to 1[105% scdnan poi(yanetbIJJ]suUoillai:e . SPS). and the recommended blood-to-medtum ratio, is 1:2.5 to ];10" depending upon the system, lli:8Jmples of these medla appear in Figures ,,,; .. ]1 to' 2,~J4.

~11-e;pti"Ghk I~J~1lQ:Ui1C blood culture baltie, (BBL, IMiGraliJiolog-¥ sw.Wm.~ IC!OCKep .. vlllSj' iMd.) re,[Dresenl:s a J1o.']"'IJJl~o.mated b~ood 'OUJtbU1!1l fzyii· !:C1ill .. EoU les co;DlitaJJI'l t~yP:!:l{,: SQ}' bmth aInd d!ll,illQar illldle- ~"ddle '~v1~h.lfufiOO 1Igat'· i.ypes: ehecolate, -_Ji.u;CQrnl-e)y; amlliLl~.]l., J3Jooo is Inocl1.il1atedinto the bot~t,. at the tlrut:' or coJjJ~tij}1l andtrensporl:ed 00 'lh labm",!'rtol" .. ~JrI ~I hiulID,gic~d 8tlf~CY cabinet. ,an <J!gaJ slide (iOOrlle ls attached to the bcrl1:le. Inverl'BiI, ~n~ h:U::UJb9tOO. S'lIlbc;llilluoo;s. ~e ·pertol'.Jj]~ii!by 'hl'VCr~lng 'tiJ,lme bQ'l!.'ll1e H~i1f1. '~.O lilQCul(lLe 'btl.!.'! pa:ddle~

2-'12' IsalBtafl" Ilys_s-centlrff'ugaill'Q:!II1 IlJklQdla.I'·, '!l:U:F!I!!!i ... _am, '.6 ,and '"o .. ml tuba. ,w.ampOlei III..aHR!lliorle.~ 'CrBnbul'Y! N .... I.) ,HI:B reco.nLlIl~nld ~d ~al' the ~:goll"d1,01ll o:Fm:v(!o!)bm::tcrl~ ill1~ fl!.il:ngi. TlJe [uhecoIlJlmns 01J bLood eel[ ly:sil1.g £I.l!liid ~f . 'Ej:i()mC'll. l'olJ'P'oo]Jylene, S:Il?S. ~UQ~rl. andll EDTA Spe~im,ens collected w, die ] G-mltnhe al, oe:rn.tdfu!:too tLI. 3000 X iI, ,m 30 minutes, The ~~p~ant is dis '~rcled, and the sediment is' ~~i;L~d CUlito se:1Beted media. SV{,{,!'Il1~ns coJ!h:ct'ooi U1l'I ~Jle J .5 nlL~ tubeare p~:a'tmJclin~dly on~n media,

2"'13 BACTiEG t!laaCli Ci:lllan!! BOflIeS; am 1~1;j:n1Ll- 111'tctufEIll 'Wilh a enok:e. uif tIT.lccihml. sl:Dch :IS 2'11, rim:. :2 { jJhll';:.,;JJI]~ ,~'erl~ P~~Is:.. shown h"!re. The hettles r;J re used \,\'1llh tile 9tlOO I~mlre..~~f;l.1l .seI"iIe~ lnsteurnenr 1111~)'~}l~'-t Hcc~on~ mckiltl:l"(l!'lI. S:JXIr]~, Mit}.lhe.se ~lci]~" are iI1re~ded for ILC(;Ov1;r~ of ,~~Ctt'~Me ,ilIIr1d :<tI1!l,!Ili;f(tioie l'lilicr,oli'lr~i]jSm:s.

:~-1 =s lS'ood colhl!lc;tlollll!!ra:c .... gh~el!" tUbe!£J. Thr8t'!Ib~Dod e()I~eelwili vilcll!mim!!rllJi:les t1~fIi 10 rill'P,IO: S:PS-oomaal~l:i:ll'Ig, S~II:n, ::i~p~~r~ti:or l~ [11::. !~Ilcl 3Ilcl:l,1IJ m ,hqJ.E1 M:n-ct1J1ttri nfillg {i1t'>Cmrl nick tnsnn. Cm,kcys!.!'iUe" M(L~. The pi asma (Dr serum dh!~111,(!dI3 used for rt.h-c ::;crodl,~~()~is (l~ tnfeeuous di:>t'<ILscs.



-_ -- -

2"''14, ESP BOA, ael"Gili:llc, ,Hndl 80N anaBFt:llblc Ililaad cLlilllu'l'e ImBcllBI ~'O be: u~'led 'With ~ln{l W~P auto]!tmledllll(] nd cldlulrc syt;~em t i)I(CI)L::thor.urnrles, De[ffJo~l. Mtch .. ).



CHAPTER 3' Specimen Processinq

II here are a number of important steps Involved in specimen processing that

mclude (1) entry of patient inftTFim@JltJion inlto ,~I, computer system ormamlual! log: U i vlsual ezamin anon of the specimen to determine if acceptance criteria are met: (3) SpeC[ffien preparation, sale t~on" and tnoculation of media: and. 4·) prepll~ mUon and .. mlcrescopk examination .of d!~rect s:m.,eal' pen !t'eceilliP~ into the lahore~ory; the technologist should verify that the specimen is labeled with the appropriate pa.u,ent il"l(Q,rm,~tlo,n. U:tl.btJbd,ed and. mlslabeled spe imens should be hand.Ledl accolf,(ID~ng to the written laboratory po [rnc:y. Any additional patient information needed to process the 'specimen should be ret ~eved. rlll m the labo aJtory lnlormarlon system (US) or other sour-ce"

~-1 JB' try, r ~~1l1eD 1.lifof,Wli'lliOIl Inito file .c.mnp ter system. U ponli receiptnotn the lahorn'ltOry. the speoln)e!Il shotd(IJ be: entered .i11l[0 the FJS, or m.anlli<lllOlt ok. PMicn.'t lITfOmiH1!:iOD. .inii';ltu;lil!lg d~~gr!os'IJj: ~m] 9J01[cmlcrohil81Ibi:l:r:'.e:P'~, co:n.lplere s.pec-o Imco infuriimiatJi()1l inCluding lruIl!ltomic co'J]~tio:n 5100. tlrrw of collretiolJl, and ~est l1e-~ue l'S. and spe, :iim.(l;.tl ,mx~io,1l number ~Qt!!1ldI be: 11. aIIl<dde befuEe S~imCil proo~i]J.g,.

3 .. 2 iExamrnati.o.n 'of tlw specJme.tli. eoatalner l'.o m.ll.firm t!I_;m:L the peeimen is properly labeil.ed" t[IJ criterion rQr ,m.Ctq1~i<I .,00.. i protocol for ban.d.lifJ.g, lED.l~beled and !liI!i:<;libe!~ S]Jscl.1l1eril:~ should. be G.ll'i:lil.Elbl.e aad [Q]Jowed.

3-a 'Verifying that '!jh.e i nrmlID a non 001 tl :,spoclmen cenlaurn"el' label c[lrll1~J.XIl1!!d81io Ktb llJ:le: l<lbo-9lory reqUESt rnrm. 1m.· [lro"Ji:le[~ l;JIooloo spoolme-ru.s]];Otildi 00 proeessed only ltr oorrecl;lr~e,Old1on aad dool.lll-nel1lJlali.oT~ aru olJm;ple~C',

3;;& E-~millD:l'J.m& th,e speClimem ,~.UIl[!'IniIller" fol' the pre-sene f ocollldlrs, leaks. and ,umer poteuJi!l<l:l.l prcilllema_ UQ:Jlrcrs:~1 Jlrecrl.lJ~ l~mi9; 1:I11011lM be used w&cn tm.mlliing tlilll spe ~me:ns.

3-4,alrCrI.1Hy examining 'the speetmen to deten 11 e :if lt m.rnt.che . tl'i.e spedmen dIl:Srniption provided 00) the Miora.l:l!lI")' OO~l[u;.:st :!bm ,

~"fi' Ma~pic examinaJti_oo :~ 3Jn Impatil:iml de:l'e.r:mlina- 1100 for ,aU spteCLmefls:, bO'L'll:'eVer. it 1m~arlirve [0][ speo1m~' s from e:x:pecio.ra.ted S!P"I,ttPl. 'l'he miifocos;opic :appe.airallces oflhe f{lulL'" SP!'I.I.1!,)l1:Jili, :specirn.en, sbmvl:iI .. w.nhis. li~lIDre ;:'Ire (l'o,!il 'Bft) ~hii.ck. mJ"ll'C:i~ld, p1!l"mleJfit; i[.I.tJp ,I"itili~l) 1.'Iruter-y s.~U!,i\i!;. (~Fti lelL) mucO'ld, !bloody; i ~'{jt!»m leJl} thick. rrlllJiool'l center surro:lIllll:dfrl by' a1i.v a,

3..;::7 A ~\ll:liJ~'WrtUe:Dl P[\OIl.'ll~£~~J re ~nlLnll-B.lIJflll]st be a\!aUl\lbl~, H 5li:t(!!uld tndl!.~depl!)] b::jes [Of ,a~eptill:g anti l!~i:ocli~1:g !i~liBc:.Ull1en.s_ md :jgui(l£lh!l{$ fO:r':!lpecImCfl process Ill,!!;, '~hISl :!l],IUIlI:li1 sh (n1U~d be cOO1l;s~lloo for oorurplete ,illi'OrmMiGIiOJl':i, :llpo€cinwl:JJ preecssing. w~C'lUlaijJQf!! or :medit2!. !lI:rnd h1lC3Uht~.o[l and ,almOllp;rnea:i.c CO,iJ;dilim:u:.

3·8 A ~[l(l.m:t~(m uI' em:ich®d. diff!:retiiliEIi.a)['Q,u, s~J{]~I.i\l-e ilfl1,e-, dia should he n~il,1~~ie [or C'tJ~rLme. A rlaHy SLlpp!y o:f media :;!hauld he: stored ,nl roOl'lll!. '~!l!l!pe:r;:riJ.lt'e"fl1d e"sfiJy~ ueresslibieliO 'I he ml'er:.

o a

3-9 ,A, Ii. sup[l1Y of prer~rJl_]i(;ed OJJrl,1Jtwob]c media sb~'lIld 0lI.1llx1 b-e a!,lOBilhihle ;:J]j]{! used ~]lJlll)1 nn !lC!eCllerl :o:peciimernJ> m,; 0'1I1lullJ@illilltfue Ilr(lO~UI:,e manuai, B, 'b'!:tc ,a~pr(il)If~Olte rnefiltiOl .nld sll~],es ,!>boulld be wieVlec.i on the ilb;~~is ,of the :!l[JIoclmt~m sonree and (lldtw{! [i~u~t. 'll'b~tlltll:ilu1t shL'luJd b~ IliiOOIed wllh iLIJ!:i .~a1ieiU IderutifJ!ingt.ac~sioiJl n~m~~li IlCf{JITl U"l~ ,flf'l" irloet~l.t1too.

~oo1IO A S~,lil3CtiM ali' tnoculattng loops, hl:cl~1!.IrlLI~g callbratlng lo{ilpS, hmllIlllC amen g the sll[lpjies rU[P.tUtle~Jii~"

3··'1:11 ~r:_~!llW ~J~[;iLllL~IJ!S iShonbl be mlriced or Il~ID.Q.Bel1:i:oo~. b f.ore L!J;racul>ll;!jo~ onto lI.(J100i:a, 0lJ1.'il! method makes use of mr... lii~ ealpci bladea 'f11ie speclmen jspl:at'Cd. :iJ1~o a s~rlile petd ~Bl"h, and em:' IU tv.: ~tefile;S:lu'gi(;~JI scalpelsare it1S~-d ~o lI1il1Ilc' ILt· :;;pocim-·Il utliil U is, b,oIDogeneoL.l5 .in oomilisL~Il{;Y,

.. 14 ./'1. portion cl I'll'€: spcCiffiCil is pla<icd ~I.'U:~, ilfle tls~,e ~riLlidec 'I,u.be, omdalll!eME 0.5 :w aU Jlutrocntbr-athls iltl..dtcd to moisO.e1l! lh • 5,~ci.m.e[i.

3 .... ~I AmI exulil~nl ,~l~thoo fm.- pre:<--!l!ri~o!! '~)'<1iT'Ili. slain (b,uu fresh tisSIi'€' il;, ~o l;}IJe'Sii the l~tc~d1rt..,.:tly O'J.1I:O the ~ s-!.U'race. Sterile sUdesli!h.olllld be used (I ~re:s",'(,,· , t:b~ tissue for addl- 1ioll,;!III studies,

3",13 AMlJli!')'" method fiJI:' MmllJZCl1iz]ug tissue IB using, 8 di:splI';S,ible U~tle~rl ru;ll~11Jg lett tSage ];i'rndm'i:s. Cary, III •• '["hb SY.ShIillll :m i Ilj ml~S OOii~rut'i1iIll"ttGI'1i ,ilInii ~il'Jtte1"hrJIg of the speclmen,

:3 .. 1115 Tne specimen is bl..ll'l1og8li1lmetl as the o~rator IlrV" ts 'alrtd 111,1:lhes Ulle pestle i:J[JoIiltr'1n 011.0,0 the li!!Stl~ Th~ procOOI!.l.re , h.ol1dd a1l.\!'ay::;: be perf{);1"iI!rIerJill1 ani{]logicai safety cabinet


.a,~;g S-~QimeliDs w!Mt Ire '~iimliIlr.~ can,!rully berClrr-e 'InocuEm,LmJl of medm. It.. r.epre: .f.llt~'I1V(J p.ot"lIirut that has ~n abill~'[mal a[!,1~~I1!X: '(i.,"" IJoc:ks~r bl(~llld,m1iJOOIN;,. gt'anl,illt:l, pueu:i'ei~l. ",bnQJrm~1 coJo:r", er ether ~~~L1S or M1llnf:i3 "I!iImlS .PMCe..~'l~ $brJuid be selec>ted.


38'17' At S~U'll;)., as !l~'!lOWI[! m Ifl'tS ~~i!::ur.!l.". I'nu,'I'~ iDc ,cilsLiii1lguuhed rnull £p·Ill~!.!.m, Till:' im(mes~]D[1 Iwt il ~. lmen eonliilm·of ~~h"'!! is ,confirmeil ~1i' I11,i';;~.I"m • iI Gnuo, iStam andexHrn.i.l1ll:ll! ~!:Ie s!I'ooime:1l 'ml~oopic;illj. l( the wj;et',u;l;t,;:lJpjt,; leJGlIIllina:tilml oonlli:mf! th~1 l1ue ~t:lrncmt :Iil; not ili[iluttlm" ~ s-pedm~n li'hl!}uM ~ rnlect.ed and not p~ .for l'Ol!Iltine hac'Lcl'i.H I ,ouitw:~ B~, Gr.,am. ~L .. lil {I1!Wli' pOV,'Ef. oJi' 3. g.m;lIi llI,llJ~_ll~y iP,utum s]llOCim.e:ll, c.lmr=.\oi:~i"~d b t I&ck of 5.gUil[Ji!l1iIYU::;I~!i!!r'= .Ilia!. cells. Mod .of til, somatic ,c;eU;s, seen are Iil'iiCU!lJ;OPIi&, C~ Gr'!W1 5'lai.nl (Ill""" pc' .. , .. er I Ilf 31 1J1Ofi1!' 'qllIiiilit)" S~l![ttllDl SIliBC]:... me!'ll, cJ:ioBIilCIEl'i;red. by !!l!.!fill.'.rol!.l~ SQllillmlll!US epJft.bil:::lla! ~, The c.rttlD'l''C of mh .a "(]I imco.w.iU nor aiJru~~!lely I~ the dlrTitcaJ,&lRuMioD and rna:rJ1JEl1d mu .. ]eadi~ ~~;dt5:"

3"'118, 'fh:~p.Dl'tion ef the s-peOl011f[i :1]] (IS!, lIike'ly Ile) Ibur:bo:r Elle jJ.lr~[;jjJ]g .ntli!em[J[1!,'o':ml~!l::it; sel~~liB~l A:::;.::unple u( this s!l'l;IIlmu spec.1111l.m:u :shQlI~.d: be taken rr.CD[lfl U~c ccntrnl. hloooYP(lI;Ui);l!,

3",20 IPw.~r ~1il~hril:iqlJ:~~rn,tIinlll!.·be used \\II'bie-:n Fepariu:1g clj~ rect ~ll1etmi. Wiler!! ~iIlQClIlhdil1g a~pecoill1len on.f!o a slide w~tl~ a Sl'i"'.!b, th~ 8\!'i""'!~ shorud ~e rol!red iJt'et' :anm!Jl the: sud,](;e usIng a ib;itck~m1c.1-r~h motten, Rcj:ecl dry swtll.s aml.ooqu(1;!l ~ ft. rmw ~pc.cimr::lt,

3·1~9 RQjCcti~l1 of specirm:et'l1S II'! <lit .<b:ppc.;1r m.acoosoopwMiy in be flr,np~ ;om ILflifectedoolJ_ro;:, :mdl MI'beClfJll!hi't Oltlid showlJI lr!! t[ll~ ~re, bu[ ilIP.P~Rr ·!!uHI.(l(;~l'lilble 'wner.! exmninoo mie;rl)oo SQopicrui},; :~t~l]l!J!a. be Hoolded], '['he (k.\t!U swn shCluLd be repe,atediif tilile:re is a ,Dli:E[lep.Bncybet~\!i);D the micoosco.1:l'iC e::l:a.w. aQr;i[he gr<tS~ '1l]l)1le'annoe oJ tbe s-poeolm.en.

;a:...~1, When:)!. ~leri1le p,ilStmu' P~I)!o1.t(l.ls I]sGd '~o gJtlOCli ~mc flu" t(]s i")];['OO a slide, ,oo(i~h S[:ICChiIWH .iIiJi.J:JIit l:ieplilll;oo m:i the ~Iide lD rO[m iii. elrcle (if <CI[)p.roxif'El811!'.Iy 1:;: l;nlllli iirn dtameter, 1'tlie ~ilicle should! .til!!' dry :irn. [me billlog±C:l~ s~ret)' c:Jt.Jlnel: bGrnfc·lt If; 6Ji!oo and slaimil.

3-22 lt ~Hay b~ nl:!t"e!i:'!!Blf}' La use iii iuup wpre~<l.re!lic sllde Ir t!t~ saID]}lc is f;11n:JI1IL I~jl.]mlgh spoctimen should !be d(lp(Jsil~d elm the sllde t.oft:il"tl'll :l dN'lo· rnf ;IJlpro:;:lrt~3tllli1:~TI :imm. ill clirlillleter, C~e;}[IlIJI:l ids, on the ofller haml :shlllllld be clI~r{)sil'Bd. .i:1':I ~I 11I'~81ped .;I'm!') <IiInd. Hll.(f~1V.;:d to ~if dry, JI. Ci!I:OCC I'Iltr~~uigo' i.!lI:':oo.1lfl1- me!1ded I:O\r :lilUl:Ik~ug ;sl.'l:IClll[,S of l1Qdy I1nid_s.

.3..;.23 A smul l dfJr~p ~r a i~quld s~~mcn 'Is ffi)IIiliCOO U~ ~ !!'il~~. ,1)11 a Slicl.i:l and ('~IlOO i:m.nlel'ilaoollo! W:~th <li ,Co\ll1:f !lllp .. ]'il~s: "'~l!r,J,u~ll:wn ~~ e.~~ln]inl':!jj uuder X 400 P01\:'t!1' 'Iu tiiGl®1:l bfi;CW['iall[~!i)lility; motile pBHlitlcs., and fCilIr other 'iJIilvcSl lgauons,

3"'84 A" rr ~hc SIJ'i~,dllT:lell 'is Or.1 ~ swab. the 3¥H1l' '!,IHU~!; should be ijlj)"~ [,rood 'l(}i' mlm:ingl he Si'ill'nh .. crpss~me {]lmd~ [';fUJil. of l:bepli'lll~. D,.1lf Mly ;;)fIIC swab Is ,':!v,.n=lbl"" it ~h!l!IJkl he !j'Qrl:c~a. in asmall a:m.mmJI of bOO'l.n. (0.75 D.I'..) 1,0 rail) aM, t~]C I)!.:sul~liil~ suspeasien can be lU~ed to' in.Qi.~l]IH~8 plates llIillld preIHjI'~ the ::mll;,lIr, ·I,lu[~l=xlull. sh.ii,f~lcll be ~rll(l]"imed UI1I a bl:ol\ogi\cal 1mhrt)' 'I::abine'l ,

3-.a4., conrl~., Ie'" T'i:!.t.'! JPlnlDaJi~lil]l jHm:u~,~ 1'1111 is ~lfCaf.l wHh a ~loo-£!" Uj~ng it! back~<lml-f(I,I':lh IXI~Jtj;Qr1I, A \~1'ire lnop ~hould be sler~lur:d 1::i!:l'l;\J\'01ll1l each Slu:IXl.'lSivc qUl,m'lHlI'U st reak,

~3·l2e StClftc [()n::~p~ m~ lI.:li-1:\IJ to PH!oeS~ hardware such <1$ c-atheler. !,ips. ~Whe :liDn::eps am fir8't 'I[l~rl ~,(I ro~1 the ,C<!tllh,eWrli.p lJ~er the entire surfece or <J:fI! Oll,1iil1J'plaue. and then [0 phtt-u the c~iheter lip' i!ltD 0], broth mc-d!urn ~!ih(llwn).

a!1-27 SpJO"Clrrrlell!s III ti"Jtti riI~ sl'lmi.!Ci be dl~otl r:aged~ 't'i~ ls ure best iuoctll~ terl :!It !.he ibeclside_ :moo(i. cuhnre 'li'irus tii'", uscfull ri"JI'I11i£lII]f ~m~y ih!M~.

3 ... 25 [.f ii:lcirile 'b-od!y rllJ.i~s :JI:re t.~IlL'emrMiedlbt' cen t:rtfll~'llion. tile s(l~nm]mlJl: sholJ!d be ~t5;tllrH~-.cd ,oiT ~h . ~~dililllCi1t. anti ~I :p.dsteul"plpe'l:iI;:e :\lh(!1~!ldllbeu~Co(] to mlx 1l11di than ili.Dc!J~~f1le Un:: scr~ime-..Ilt 0000 medtOl iiRld :sIMJes. VO]Ullrlail loa. ~1.iIll~ln tebe C'elltrirIU£»(1 should be itl.UL: lJ[i!I:~~!.I. dir.et:Uy li\lilila IPI!l-eltJe.

3-.211 A ~2Ir.t~t.y ,of mrn.rup!iJ;~['iC ,e-O:fitlit~OI1!l i!ljJ]J~' i!lev batij)'.,!D iiCU:1pfralt!1OCS should Ibe ~v~il!~b1e fQI'" n'IC'O'Iirezy C!rpatilogCltili,c ~niC:I"OO!!"g;;;lIllsrniS. hleu~EUioli t~mpel."artl.l~S; shQuld W!bJufitl i:ll ~ea:sl 300C. l5~C. and 41"(;, a nm:tg mill! aercbic. anaerobic, c01p:nu.d.c,. all(jj 11l1i~aeo;rup;h.ili,c ooml~,'I,io!lJ5. MostrD1[.l tune ,OtiW'l!lbl.c; cultures ShQlJ.l ld be U:lclI.b<J[t,·d i:l!l 3 5'" to 3 7~(' '~n ~.! huitrn:r.d <It _ lI!llosphere or :;% to ~. t'O~. 'E'he:l:efnpcrabulre, COJ: Cl(jilh~rnl. aad hi.lllui.d:il.}' ~iOl!lld. he monitored d'al~. nlc CO~ CilJIll.t.cllt [S mm .. su[\~d li\,rUh :.l g~I:IJ1l:e cClrltall1lng r:F~6re {shmN'I!1I~.

3:-29' l~le 1:e[([lpe:raU~re, h1urnidlty; ood alitllOOJ)iI£r.e i[llLl!lt 1Jt:

FeOol)deo dOlli~

3l-31 Preted iJc~d aflacrobilcplatG".ll C<l.illJ~ I'ru~d ln "ml ~una:l,:lr· (Ibi~ IWllii[!!tI1Ir rur~ 8horl.~..£ b€lt~:r'C!ll!ld <.!i"OOr lniIJCil1~<lti~[l,

~;..3~ IUl 1l!llll~1i(I btc at l'IlOOptl.Cf-€ canaiso be m~GHi.!Jt! .in :ittIi mtfr~t'{)it;i!i:" ,pt)Il!ch '[hfll helds OJflliy Hva (I~'ilI~e.'l o .. iill ~n ,(1 rl'tl!f:r\!Ji'.I!c i~[ rut l~ln~~ ~ r mere r,I;I~~'

·3b30 .Mi;l(llli~ ::;I:mu!i.d he incubaled as ~(lQn i1:,i IPorn;iWl;: ,[ufir;:J; iu:ol[lJ lati!:flll. As ~11:{m'll here, ilJ!(lcl!llaned media i:s; a[I!.';:IIlJ[Wd in an Urollf.!.)' LTIm:meT ~n ~ni[limi:;:e ~;ri!rlJrll. ,.gull ~TIld COIlW;~illrllil:m,

3",33 The G[15P'dlk m1J!lerl~je jilll:~r.~·teli~~:Beor~I1r:11 Dtckmsen Ml:croblulogy S~rslE!ms. Corkey!wUle. :Md.~ CO!lli::lll1(l;";l. hydrogen iiil.'F.iloCllCOl gen~miol"el1ve.~npe,,~ dl!lpmmblC'n~efhyl!enc Mw:: tind~C1IJror, OlIfiO ;J c~tql~till ba~ket lnrh c lid,

3~34 ThE> G;i8lPak ~l''HLBl'ubk 1]O!JJcl"i (Be!tlu[I mcl::il'lS(II'I 'M]~ crnbi:Dlllgy Systems, Cotk'il!}''!illftUe.. Md.,. lsbuaed em the !j:m'l:iI'IEl princ:i~le, The b,'dgls (~My~E:1"I imp~~meable.al1id th~'sli'8Iiem,()oo. ta~Il~ .it'S O1,W ~~.~ne:raiib]jt'l k.i[, and. co:~d (;!'I4alysl.

3 .. :36 Il€oowr.y or some r<1i)I~d~oo!i ml(~:oo,rgalll.:l!lm (I.ee., lll!.llrt!'[:'!cl.~t~ ,iP~rr !I:Ii~:I!, Nci;!l.lfell"Jd 1fQ!,J()rrtr.o.L'fi!l!) rUQuif\~ a, mJcr~ltl!rophilk !jj,[I][.!sphe.r-e (!tfuereflied O,l!J [but can. be erefiLOO! hy h~~rlllin!:1 a candle In a c losed ,i~Ii""

:a.3i15 Fm:'rtll'm'1';r.y u.r Cmi~]J~JloJl1'1d!lft tl!,~)P.. ~I [Dilcr[]Jam-IJlphiluc "'ltmo.c:.pherec~liI;D1 :tllso be ereated n D1 (J I_K}L:M::,I'I {!I' a j I:Iir,

,;:;J'·'a7' TIle t€'ClmOLQ~ist Is pJ,OIJcm:g OB 'blood culture botrle tato th~ BAC1'iIEC 9240 F.ltn:ol'c.:!i~lJit a~llomatoo blnud eulrure ~srem (BectOElI Dk-:till:J:son IDili8ignoSiilir ItJIslrmlmefit Sy~t9JjS, S~-ks..Mcl<D. Each OOI,I.i.8iis eril:rrcr8(1iI110~1 eell ~l ri h~~nc'U!tm[or ami mOril i~flr.fd e~Bry .~. to mlmJlI,ES byr a flil;olneeJI r~e(lb:!lck !ifS· tern toa computer.

3",38 Tile i:edhI1.nlo(gjs:t is IlJoIOtcin.g .:J ib~Of!d cIJ:ll~l re buUle into U U~ ES.'P ilEl!d.rIlUJ1~nll. (Dil'tm l~lO[lrllil~Orr85. n~ lrui t, M'idl, 1, .it (uJlLDIccl.ur hi phlccclf][!I:o the lop or the: b(JUI~ be'lb.ce iJrlCllb~l:uQn •. &Jc!JJ, blJll'I:l~1il ls entered b*;J III cell or the iBr;~I~)<II'i(lr ,!'Ind miGll ito:r~lle'liery I Om:I..b::Lu:res by <l pressure golC~lge lnterfaecd 1:0 ~ OOtl~ptil.~i':.

~-4Q Stf~iu~1 smears shmdd 00 read illll1l!I:c.ill,Fdcly aller :>I~1'illing. t\!Yrwrlll1l] results mmll I.l~ Ile~tlglf'llJIIle[1 ilJ!1lllttliH~Ij.' 10 [~ phy~k~Iul;lhe dille ,~mLlI:i:me (Ii the C al I, must bedm::l.l.r mn.illl'l:l:erl T.he ~J'[le tar urge!]1 resuull.'ll il:~atr~lili're a 'imrhl:lJrepClrl: ~I'l(lldd be '~i~tcd ln [heprooedlll'e' ~'l;mu.:t;;d.

,3-39 lJif.ee~ ~.lIlll1e.::1irs sheuld be mel:'ti:i anol .. fiN!2d~n.[l sla~HD.d II s ~.Dr.ml. as f.O..~lble :1~ol!'I" the :'iHde!~[Ii~parCa <Ii!l rI d~ed-

;~h4'1 S(pGdiffiTWlli Ilrl!J[;~:S~!~1!: sh:{!ItllhJ t'I.lSQ I:Ild:utie Ii 'Ii~u'ii!lY ~J.r m!1Jtll dilr,[!d l,e:;;l:; Ulli!:[ can iilS5isl, the p~l~lskL!m \'d:t 1:11 ~ me patlent's ,cti1~gnfillis." Rxam[Dk~il lncimle dilrBCI 311ti3ml rests, !;1,1,d.' :lI!l, a ,cr}'Ptooncca~ ::I:nl]gclll"tB~ ,agg,1ut I nation test :'iiUIWf]ll1m,

10' . . I he' Gram stain Is used (111) classify lIill~ro~r:g,an~srn~ on the basls of their IGr.'l~m

,_ ~ sitaJ]lling ch:aJ.rac~erisf~cs, sllz'e, sbspe, and. arra n gement or cells. It ls one o~ the

ft:w tests in clinical mlcro b1o!ogy that coo. assist in the rapid, presumptive dtagno,~~s of an ill tecttous disease'. n ,is also used toessess the qlla Itt-y' of the cltntcal spec[men based on the somatic cellular content. Bacteria and fun:gu s:taingnr-an.'ll-p©,$liti'IJt;'li gram-negative, or gram-variable. The gram"~'at1ah[e appearance can be due to aver: .. or U1IJd.er~decJOilomElltijcn. age of the mic[ool'g:~]]iJsm. lnfluenee of annmleroblaltreetwell t and other faC'Lors, ~l'be s~aiID1L~mlg re;ac~ion :is ,depe'l1de]]Jil: <in the m~crobrua~ cell walli. compositlon, The tigh~Jy eross-linked pepUdoglycan layer and the teichoic ~ldd founrl ~r.rug nun.-pos'!J1j.ve microorgarusms C31Llse them to betesistant to acetone-a] cohol dec(doru~trnon., 'I'hcy retain the aysl;w, vloid stain and {IJ_fC purple lncolor. The HpIJp rliIJsilcchari,de~ri,c h cellwall of g ram-negattse I:n~.croo:rganism~ is disrupted Illy' acetone-ulcohol and the cry\Shdvlo]et ~e.~.$ out or nile tess tight~ cross-Jint:;ed cell 'l'LfaJl!!struiCtur,e. The ~arra ntn counter stal n carl then !be seen, renderlng :gr,ainl:-iIlegatlve organisms pi]nt in t'-OJor.

The size of most bacteria ranges from a large cell OOjllU to' 3,Oj.ll]u Inlength) to a small cell. (lum to 3~m In length), The shapes of bacteria are usually d.e~crilbecl as tCOCicat b~H:::m,my. or coc.cobacJHary. 3.IUl the arrangements as: pairs, clusters, chalns, branching, fill[JJmentou.ll or c:(lr-yne:{orm, Yc>a:st C1;1!J], be seen ,ID!Jl single cells, uftenl.r'l!dth. blrndding :;'l[1JdJ or hy~p:hal. fDlamemts," M!.croorgrutlJisms 'may he seen Intraoe[h.li:arly \VWJ~hbl. somatie eells, S'ta.irlin;g dl;~lL'~cter15~lcs orindivW!lIJ~lb~c~erlliaj cells ru:rllJdllloe bipnl,a r, headed and tr r€)gul.alI':" &1 ds Inay appear rounded! "poll1'Dted:. flattened, or ;S:\!'iml~.en; l\Incroor~'lnisn'lS other than bacteria and Y"elast observed. in Gram S't1lli!hlS, al.Mlong·h they are better eharacterised wilt]] other stains, ,ElITe Tr'l'cIH:ni'Wi1:(,I'S tropho~iites, P.th~'M~i1Qcy.S'tls (;~l'J'Jnf,.~ cy~t"l::, 'flJl~;::)f111!'i.~m" .'1,Oi1ltZH tropheaottes, and StrOrlg,11J{JI~dfi~ ],f)JI!"V~Ul.

S:0mi1lUccels can also be seen ~n a. Gram stain, best obseruedwithmethanolijx€d smears aJS shown helm, IN'hite b100dJ cells (WOe 1 and f;lp]Il~(;jnal ce~rn5 tend to s,taw a pink color whereas r-ed blood cells I IlBC) ,S!~pea:r 'tan to ~ufl' cok)re<i, 'Ihe p'fesence and rough Cjlll.3nUt anon of somatic eels ShDI!l~rl be noted when interpretin!!!: a Gram stain.

Gram. stains shou ld be examinedunderlow ]:D0'WeI' rnagnlficanon ( lOX oil)jlectWve) for the qua~Uy 'of overall m~m(ng, the thickness. aad for evaluation 'of somatic cells; micflomganm:SH1l.s should be observedunder oil lmmerslon (100x. objI6CU\re).,


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III '11
I I'

• ill:' I
'I! 4,.1 P~),~ynlW.prl.(~!UHJI ~r lG:lfIku;;;YliCll fPMN)1 and graul-posl'li.1JI8 I!IiIJlC(!'l''.!ih~Jpt..od C1ij.lIQJoot!.c.i iUj;g0:li1.i,'!i\!l:' or ,%r(jl!UItat.'L'U8 rN~eijmottla;e I x 1..: Sff),. 'file clear ttT,OO siJJ]"l'olmdmg the b3'ct.eriJd (Xlils is 8!'!.l~Ii\l-e of aCi~psli1!e'. i\ .t'dp:iddire.cl bacte'LiaJ ~.1i_ge1l test m~ be perhrnu!rll enthe ;t·~ifiic·~L~ S~1ilfloo[l to ilJoll.lll'm the: organism iikrul.licmlon,

,4-3 Orn1i1'l~t'l!Ol'ilthr"e [:ocdl in ChaUll!l resembl ~Ili~, ffi.reptoclXl;:l Xl.HOl,

4 .. 5 Rl'K: and gr;mn,po5iU,rc i} !Xi !tn t 1i:r:ads. (X] 2~' ]1. CU1X:1 ~[re ~Clim~vib.at 1.aJ[~e, lrll1r:',gl,illaf, BlOtl "I)'~y 111'11 s1~ :lil1gge,'ltbi'T~ of r;m:l~lB[:a:;;~IJ!liIg,tIliw st<lJl1:lj'lomcci, ii!','O('occl,rs, or St£Hl'I"t:G1Co,"· n __ ~JP •. ' J:tqNJ'ylcco«tis ,ii!,um'8 eells are' pniftJr:.I11 ill siz and, ;;Ip. r,lG'ar,[llS cl~lSlef.ll of small cucci r -1 ,j.I.!ll iII! diameter).

4-2: 'PM!. red bloed >celL {RBCJ. and gr<il-l'lll,' .. pnsUil,."C 'oor;;c11ml palrs,

4 .. 4 ollitall],-,posmve cocci io pans, b:;lI'ili1S. milIdIDltcn;; refleu.riJ,ling s~~Mp~_~}'luc;o.L; if (X 12IJO)

..... rl:m ... ccltl,lll:al~, nr.te\reuly stail1,ed, pl:e@J:i1lug'p!:dc. round, and grm11l""atlallle cocci (Xl2:50 , In ;s'1IJ.dJl G:t1.CS, the gr~rnvadalbl\e' appearance may be (,]lII)e to phagm;:1I:oSili ood. p,m:iiGlitiJ deg.rada.l_il;lu of the IiLlil.;!-OO.r-gamJsms by tbe]PM 'r,



.7 8.11' "lnad gffll1t-positiw' l"OCelll] p'alll!:'~, short and l()rn.~ ehams FI[)1iI1 du:Sh.'.D'5 :!lU~Slhrf' or Julyei! staplla;yltoG'(l!('ci mllil , -ref,ll(]mcci (X ~. 250 I.

4·,8, Vaginal squemeus epitheliml ,lib: wtth medium, *itiil~ll :ri.1JnJ-rU~ili\re!grmu-\i'ar~O!ble ImeilU W.IDh rounded or blunt en~s, some iii:! chams, S!ll:gg,esDi!,lti' <lilt iac:wlmd]ll ~X i2~IJJ'" ,ei- :Ii l:im:,k,gr(!rlllmj l:Litcl sharp eprthehal cell ,ooges I~'J! eal of parlcru1161m \il,1th hOTlillHl l.mgllutl ccrcttoas

4b'I'I Pil.i1N and 13I'g~ g,fam-(!oQ".Sfittve b~dlli ,I x12 ~ I'. There 8ppenl" to lJel1i.\f{l baci IH that 9~, firregular In :;fiape; and ~ta.tll- 0]8, glJlilUlI1g the o;l~j'''U1CC Oil' rocc.i in ehains. '["he beaded ap· pc'aTIJ1IilIOO'lllJil]' be due- to the presenceof fit/lO~'e£. PMliiai ~t!S' riel'! by Ll:ic!: [IlAl[N, or because tire snuctu j. OJ sJllal~ piece of fungal hJIPI:! ae, whlcb. &1l atllS iJJ(ICVCW:YO with Ihe Gram s~,;]Iin.

4 .. ,. RJaCand, small to medlum !,trfllll-,IJrtisitire b[!cilll wtth p1lilis .. (lja:g aa:id: ill]l,l,uJLHlf W"nll~1'I1{l'l1~'S ~x 12 ~'[J~, SOCILIe i'ip'pmiir coceohaclllarj, dipblbemidiLike., ",!~d p'ko,a1orpillc, w,hh::n is ,SiUQ:I!l~S!lhle [iI[ oorYl.lebaterla or I !:;f:t"rl~r.I'I:W~ID~11 Oge! "e~.


, I

4",,110 lli:!i'inl:egirHtJerll '\>\'BC and plcomorphtc liu·!!.e gra_m,.=posili~'e IbaclnU resem bllblg, Clasl rirtrwu i~l!T ,rjJ 1'9!'ll:l' <or. ~·.;u'~~.Bilf.fjJf..r,I\,.~ JlP, (X 1 ,]'5'OJ ,

•·-'-I ... ·~.,,· ..

p;rr -..,- 'IP"'1.. ~

~ .. --.-. .....


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.. -

.... -



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... ", I,

...,; .....

. ~ ...

Ii .. .:..tn

, ..

IL - ...... I,,~

.", .

.. _",

4-11.: lill;g~, ~[,HlIll~V;;l.Iri~lbIeb;u;:U Iii w1th s '_ 111 'mm ends l X 1 ~ 50). ~011Ul cells aLJ~r as drurnst ks. sugge .. ~rr 'C of C.l'a.sl:rittirll'll l:1!t'(ml.

,4"'13 l".MNmd ]Ollg. UW, b~'adlild, br1:l[Jj~!n!ing:.. "gl.IID~re~1y s~Hlin~1 :8J,"~~itiw fi1~flJl'5 :;uIDi:~tivt: oF' Adiffcrm~""lDr Nocanfj~ :spp. '~:Xl.250). 'Ih~~I1iI]~ H~Hy be COIIJ!lrrn.e(l 'w.lm Ill:

Inodl§~ l[pmti~I)lIci'{f..:Jj~51l st~in. N'~.f-d~:t>! SflP; are parUaru:y ach1·J3sL

4 .. ',4 Rnmo1rnjIl~ filarnentm.!ls. slender g'lrnm-~itive budiUl . . If ()-bti:ijIled~rnl1ll. al!1la~b!_c 'GO~,y" ibis m.o:rJ.lh.u~ogy Is su • .". Ii .. e of !k1~"'i,1Illi~ :!:Ll:P, (::<:.125.0). i&'l'Obl'il !1Ii tOOl\Juy o:F iii. mi!::iI: HIIJ!.!Ie:robe, ~his ~{)irpl!IDI!lIDi' oog)1iSls ClO8l!rfdll.iij1 SPP,

4 .. '1,5, OiIfsJwlj!.irl :s!!!p .. r~gmmt wt'O COC'{loid .ftlr.Dml s:iilerprn. longed iIlOOIb,a,hO(J:l, (X I;?; 50),

~t; r~"I·· .,.' ~ -~

"., ,_ a -". -1\. ..,.-/:. ;;I' ":t .. o.II''\lII! 1""'"' -

I .- -. . ~ ,~",. , .'¥ 1'''

, '1:" .. ~' ,~, -' II ., • a. , .... """'" t- ... , ,jI! _, '

~ '\\_ '! • _#-. ~ L ~ ',. "11 ~ lr • "'-

'Oil' '\!I .. , . 'I '.'.- t. , ... 'II.·T. i .• :;. .- .. '.' =". ' .. -- .. ~1" ... 1 .0 .••• ,.. .• ' •

. ~ ~ ~ ., 1 '. , •• ' '_ ... ,. _. '\,

.. ';!! N~~-" _ ~f' "., ~"" ,. ~. '~4i

.. , . ,j · - ~' r -~, I .. ~ . ., .Ai..

.- .. 1.=-.' -. '. ~iif! ~~'. ~.. -J. ..... " <, '.W~' J!. ','~. _' '.;: '.~ .-. ~p., '

-; . "~'.~'~ ~ 'I,a. t~J'.·, ~

'- ~~,~tJti-~~~~ ,.)~~- ,~~ ';,

"' ~.' " .. ' ".~" ~.-"-.'~'.' '4;JO;.:fi·.' .. "., ~ .. '.;,I .. ".'

>14JSt~;~ V !:"" i~ '~~';l~

It, .' ~ ~' if "'i ·f .J(- -. . " "' ..

, ,. &' • ,ill. '. ~.. _. . 1Wiir,

mm-'negsJMve InlcroorgaJnlsm.s can. aJPpem as rncd f coccobacilli, and baellll .a Coccw Can appear as singles, in pairs with flattened .!lIJdj'a.c'ent sldes.and in I' lusters. '0'-' ooh~ci~~i. :cU'e 'l1S11ally small to, medium ill sim. B,2id~~i canv.gJt_ry in ap'pe'~_rnDae fbolU Slllalm;. faintJy st'awnwl1g rods 00 hll'ge. p~um,p rods \!Vlth bipola.r staining •


.",,8 Rile aJnd gram-:lfi.cpt:i.~re cocci ill :sil~gles and [I~~r. The Hdj~'jJ~nt sides oft}.c clip.lococcl apij;leM nathlllll,t!;I(l (X 12 '~O}. Theis miC'l'OO,rga~'islll'i is 1 ds;"riil .l.!. r!II !JllklIs shlatll~d r ... cun .a I:1k1od Ct!Jltlillte broll1.

•• 8. ItO and small gram .. n.e~a~ive coeceb d.lli .in clusters (X U 510]1, This: !'Ilicroorgan61l:ilJ, :staHlN wm, the blood GlJ!hare br'Othi wus i.d'cnUfied mBi'ncelh!' spp,

4,,"7 lJrelbr;'ll d1scltfifge with rMN aJl~D iot!rm:ellutlllJl"gralIDfie-g:~llive dip:]MOQXI. sugge;s;Uve or N.rj::iS~Jiil !lo,m)I"FliIj}(!ill! ')(1250).

.. Ii!


, ,

·4-'8 ItJl}C and gralD:Nl:egaJI1,'Iffi ciXiooblldili In pairs sebseICJiiJierl~1r}' ldenli fled as ,c.i'ttM!m.d,,']I'· PPI ill a blood cllltUJfC broth

X 12 50).1'h~ JlWJ'pMI.Qgy IL'e:8eIDbl.es. j~r.i!hrer."I :S[l[J<. ;md call be casUy rnnSl'ltiterprctM as gra_m-,' egativ.e dJ.plol;;{lc i. The HJ\CRK. group of rru;tidio~ ~~m~l1cgaH\fe bu.dlli, (HrI~~I!u-~ii,rls. ,lel'f.· 11 .. ,OOCWus, Cil",li~l'n;1t'rjtrm. . ;k~1 na. and Klllfjtlk'l'). often, detectedl as !Jisen~;s ·of ci1doc;;u-dlil.Ds, coin alsnappear as SI1I!laU iOOIQ" .OOD3CUli,

~ ~
... -'
• ..
,.. :
.. ..
.... .-


~ ..





I ,"

41,.2D R13C and piump ,gif3111-negat!Vc 'lia.cUlI '''~fith bi[(}Olar litHi!li'llig tn a bl od C "Ih.ll'e' broth, !lt1I8....~stivc of "cdJi¥ l'1illi5,." beloog!T1& 1,0> the ,~~t~robra:'lfFjtU'M~! (x 1150), 1 Le shaet, thick [bacilli. ~Ire cI:i3ira.t.:ter~tic oj' ESI . .II'!f'rid~~~ colt

," '"
I , ,.22 Poot1y I. iITr:lll~nUated PM and:lifew:lde:r pieOI urphic iI1l rem- J:lguUWl badlli ~1rJlt'Sti~l.e oJfHru:'\lI~j]r",JI.i.'~!~ spp.. ,01' i:lfI<l.!!D"'· b~(XI250).


4·211 RIBC and l fig gJr.:nli-nDgtlti.'ll'e t~!HII w~lh rounded ends iSllllgge~. e of f'rufrllS ur P~"t'ij~IrnIIOJrll:llS;l!p. I x 12 5.0,) tn .!'!! 'bE" udcul W[E '1;.001.11,



":23 l~l."trrc:lIneb pJIllOl1lo.rpR:![,c. i;li!"lllb't),. tfl.Ullillg. SITII .-il:'-,;.~llive b:acmU wll:h (loj nted end\s !lnGgc;\lll~rc{)f f1ltsO'I~'Cf.itrll1l'~i;iiW(KI25m.

" [I

,.24 .&J PM'· .RB-C, and slig}'ntly C'UiI.·~ xi., l\;i(;lnll![la-&h;:lped •. ' lIlllJ-whn g, .all . S-Jill.aped IDjcuoor.~nist'OO :'lU!l!i~ '1 Cwnpy1.Q! £lfr spp, (X 11 'j;fli). lB. WrigIJ)~'s ~~~m of Borreli«:ip. seen in :.J. tW' ]}h~raJl bleod Smear I Xl_; 50). These hehcal baClklr1a measare ~p~ p.:r.(j.:l"~ln~~.ely 0, 3 IIATi:'I ill diameter aJftrJ up 1~{I l5 !l tn in fm.lgtl1 wllh liP' 10' ] 0 ~ooofl.i!."Qlls:.


Ye. st ce~rus re larger thau bacteria, usually ranging (rom .8 to 1 S um in s±z .. The. can ~rnJffilear as sLngie cells with buds, in duster' . and wi!th hyphal filaments, Stalnrnng of Dells can be soll'l€wha:n spec~~Jed rather than confluent.

4ii,2;~ Rou!nd budrllufJ l''''a l oells Md~ '111g1e bt",·toco.J1~idin and speckled I uneven taln i IJig !:i lilt!! lI[.d!ed b:y dear il"I,a!os t~1:.'ll.I:l el>!dlldr:d lily I he pl1lpsm:~hilj['iril.c 'Ci'IP ule) wggestii!'ill' or CJ"IJ[Jt(It;I:N:nts spp, (>:::125,0).

.. -.

4 .. 26 O\r.tl 011 dd.mili yctu;t cells and all i:;re~ularl~1 stalned p:lClWoilyphall. tihllTI~J1iI t m,ggesl:ive or C~mEUd'a spp. (X 115 'l.

4;-27 UIlC. CIlIl!!' stalneil 3, te 4l1JL1II ~\lidl1li nondematraceous Ihj'V1~;::tc \ nth parnJ t'l wall ~ and" -" h1181l. fu i og ;1!J!'Id !\ 'I't;(limJ~ Sllgg,e:slI,; } of AHJ:I't!'"!lMIIJS spp, (X 12;{]I,.

, • -' .' li~,

I, • '. _. .: i! ~-.

, . .~-' ...' ,-

CHAPTER 5 Micrococcace,ue

101 . ~1I"e>e _g~~r,a ol the family Micr.OCtlOCm:e.tl have ,been as" ·~ated ,~1\'itth inJecti . ns

III humans; S apllyloCt'til s; ,MU:roCOt'CllJi" and S, "(.mUI·tOt'OCI~. Of the e. SUtpJu',rooo C"W'i spp, are by fur the most cemmon cause of human ffin~ec~ion5" Both Mi' rocarCJlS and S'tomamcoccus spp. cal] 'be ~rJU[J.d in the envi:ronrncnt. us. lJ10nnarn UOiI:;3 or the skin and respirato y tract, and as pulhogeos assoclated ''L,ri;th catheter tips and other hardware,


!!ii .. iI CI".,aml ... t~n ::dl(ltW'! 1:. RBCs and grutil.-!il(lsnlive oor:ci Jn p3!r~, letmdll .• :md grnpeUk"Cclust~J1] !l1Jl,~~stit1e(!iI MkroUie-, ,ri!CiI(,l!i' (X U5I)U~ tlD.llif:!gur,', d~llim.iCI"OOI'_g0iiDl;r;1]1s.:lI:reS1tophyInrot~ l,

,&,.2 CalDRlas ,af S_ph,ylDC'oCiCuil'ii!lul'!I!II'Ue ,on 8~I!IItI<Ba:p tJl'QQc:!i agar.1ypi('aJ: i'I.ppe:an'l'l.Cf'Or :i.lIl~JftUS coll:lTlles I~ it>7c·amv/b1,1Ji" celorcd (..'r.l~lJIlies sUfllPurnded ~'a OOllt nr complete 13. hC'moJ~ is.

!!il,~a .A, elulgn!'" - ati CiIiHIIgU:I'aI!u,-:n~tJ" ta~ollil!!ln ,!Ii",", sheQP 1bI~lliliig&Ji':' TypiE.:<I:ll m1o.mie.<; 'Cif CNJ~I ~~Slc-fle,gati'L'~ til:pta,ylOC(ilDCi He H~l'ihell1tllyHc and \l,'hll~ IB~ C::alD.!I'!!I~ .,1 .m~cu - ,iiIii!P'P. IDlnl 5% I-Heep' ItiilCHId aglllr:. '[ 'pjc~1 cclomes (If Mf('l't!wu.:mi appear ~~]Jol1.yell(l~\~ •. Mlr:ro '~i'l'lfS SP(ii'. W!uHlly grow ID_m!"C slowly itlIan , tap~'lococd I ,omeLil1l~ reqLtiooj; 48 ,illOLlrS of rncuiJaLioni before ,r.fI~(1nles are vlslbl"',

·s .... , Bram 8m1" Ofl .Mf'e~ ~es" M1- '(:I:'Osc~pi:C!lllljl. 1I11Li:6roc(J;tci :aml~[ than .:n~Jllwl.c.c~c:l aad <lfppear. m, te1;r"cls Ir;aJther than gl<'l!J?llililke dlJlsOOr1l (X 12,50).

!:s-e F8",,8",~on -" g1!PGD_' is IiJrre [lit L!r&s m.e~h" ,OOs!IUilo'd to (IlrreI~M<lI~ 8t~!JJ!JPlJi:ctlJii ~PP" fmJ.TI!Mkr;ococc~:Ii 5Q)p.!;.·\lh.s1ill g!l'own In an oochJtim:t-fermentatillll tOP') lu.tecilhl[IJI, ~laphyl[HJ[lliXi pm]t:lJOO RtM ('feJlow ~lj)r) from :gltI~ l1mderralDillN)(liti;rhc',oo[!l.diilim'is. c:roo~, bY' rum~r~ng imirL®TId an ,o\\'~r tJn~ sw1; .. ~oo ()f the <Ig,ar (t~J[,li~ m;~ thi!lh'J.i). Tille mLcmWWii[il'lm,~13U i]ftil~21i!:s ~u!(;osc aem'bi~;I!]1y (w~~~m !iI,I"" rlwuj.

5,-5 'CD,talta:i!HlItoild. The test i~ ,pc.rfvrJIloo. 1'211' iI'Uj{lUIlC, 3%h.yd~1'II ~r,oo.:-l{l~ QH~O;:) W1l CCDW~'I'Or!1l gl~~ltde ~[b;y i!1ddJ.~ C;)io.l~J ~e 001 II 'i!Jrui}t1ml i)[i[<:k te !I.l!rop (!I] lliO.'! on II £[ttle.. ali: slwil'v.1l he:re, 1hll';~rp'~!'il:B1:';Jmlt11l!! or buh'M!l;!i bldjcaws th~l\ the eHY\me, lcattillfl~, has hydt.:l~cdI H]02 :llltoD a~gcn "lui) w~ter. Sla.~by~(lOOct! .m.d :~lIi~oocja'f!e &U~f~Fe'~lrfrilitedi frnm. OU:1G:r. aerebtc gcr.aill'li-.Ilo:sl!tl~ Cij)OC~ by it po~ti'll';e cotta1ase test t~'is4iM,',,1N1) bu.bbl~'illJpeHr W, it. n:~~b~ t~'::ii., ,relSi.!lt lhfl;"J;"


- -

5 .. 7 DluCOSG' oxldatlonl In !DiF med'J:urn. i\;fj~ ,wr;{"~ SPIJ. m;;i,tli~ ~lliclI)~e (rigft~, !jl1mh\l ,,,o/(ir iJi ~Jill 4;,gJ'1, .LtI!~~J! ~lIUjl. d!!J lI]O[ [t'!rJl].ern.l the !~li'builh1i',dnth:: Ut!JI, 'lolj r!;',ckll" cluulUI!: 19n;'lJii J jj'! th:e i111L"rl'QM t11hll')I!. 'flile add liiWo'llCtinu pnxlilJ'i;:e(I 'by 11M-Irn:OIIti!;\!'l Ot"~lliSlllS a[).P'~m:s at tl1:e SlunOllClii' .Jr:Ld gm>dlJlaUy cx~tilld.g lbrrUt:I~h.olU ills [iwdJiili1]J"

is'':8, 8ac::ltraclnl .JLII.~pI 6(1_' teat. SI!.mc~l.JtlblJIl~! to lW"1 unns of the anluWl;i!lk b. citra.cin. ~~. ;d:lliJ 1.11'5&1. to dm'~[-eLul.'~~ui.e SI~~IY~"CI!J:S :spp, l~[um .Microwal:ls. 8p]l "rtlC :-i.ur~ [itl::~ m !liM'I1~"1folmIDt"'i' :il:M \~.. is ~~u~·oo. wiJib. tli¥.;: ;[\~ croo.rg<ILlisrn, HIe d" k is appl:led, 8ndthc plate Js i:ncuj).u.,d! !Jvl;!lrIlIight •. StatihytocOl2 I Moe res.lsUlt t l IJ.G'itI!ll1"lits bm:itr,Cldn. '[lmle' (J. i.r:ilf.tlbfLioFt k.tS UI'a'N or ('"Q~jrd I I) .! 'i1im. iIlJi'1nlsm Ofr al~ ri g'Jn I w!I!di n.r1crncooCI arc' llsreptible, 'One!. oj !rrhibitiOll !J'rt~~'ler ~JI'1ii11 ilr t"qlJaJ r., 10 mii'J~ ,00'IJIInfml 011 ~'rdt;ft'.

5 .. ,9 Slid .. ,ooayullil!lIl1i1!tQ&1 PQr.1'al!'medi an a 'Bla •• ,.!IId _" Cclb frnm a lresh colony ac inc]c lined lnto salmeto make 0lJ smooth 5!J~vmlSl()1l (tr]H.llInd.:JJ dffiJi',[lf 'rabbul pl~s ma ,iii; i::I.t1i:1B~ • .r~ tfI...e org.1nlsm pIJSS~SS~ l~l!lID'!d coagulase ~ "diJ[IJIJ .. p.iro;~ f;:t'Cli.U·'). I ilc e~i me ucts on flbt1n'l.ogoo ill tliJe plasma aad CilUSCS ~~ lLlpilJi~ of the 'b:l'Ctcrla, as :'lhmvu ontbe ri.I:l.ru;I, S. ~un:?tlill::l till! mostcnmmon r;;£I1hugcll t:I.[lTI.Ol1g the catala~C:-PiJsUi\IU. gram-l'u~l!:i~ cocci. f.ll!uil ~~ is casdjr difCi;'.retUi;[ll,e~ room >!)I:ile:r taphylococci IJjl ~lrIe wagllll aile reS-t. "(]i<l;,;Ld;jlS~ is H thermo table ooz,y~ lou d pnmmill! n s'.I1'W""t'HS. 1'b~r '<lIrJil I::\.m fQ,rnl<s.,of coagulase: bound anti Iree,

5-'il'O' '1ft18 'tube 'Daqula •• 'It.llt dli!~!:!tl~ [rt"lilCJI}iil:gLl.· ~a, fl. Mlr.Mi1T~]ljSI!ll"!> are i neuha led 'ill :1?1~s<ma ffl[ 1 to 4 hours, ill1d tllIe tubes ~re t!ll"ruoo (Ill ~hei!l' sldes. iii> S:11(lv"n L1~'e. ree mnl[lolast: aets 011 pronlfQlw~bll'l • nd .lib_ I'lQgl:!1 "in n .. bbirt lJ.)l!liJlUl<l. and forms <!! Dhr!1l .Inf: (tlllt~~ml th~ lem,


.. r

.-t2 Antl'm IQI"4;JDIiP" :!!!l":tI~~iPt!i~~.1ity tellJllng lof S. ,o:iIIUI\li~I'_'~ Mc.Rl r.fJnlanunliY-<lC(jIUi(rrd Sllmin!li or s.. {jlln.-jl' ar~~L tautto pentrlllu .


,5 .. 111 IM'aninilal , •• 1,1 oNlIiar. II" 'leaJll il, l s:ilI~1 a~"1r dIffer l!1I'tj,I'l'te..s. S. mll i."l:t rrom other c<llalast;:'-l;NJ!'i\l\;-" !;Jr;;t [ll·pOSill1ve cocci. The medium ICOImD;;J,ioSo 7- 5%011 'I. \" hleh 111li_ubit:'l ilu~ :f:!wrwllJ. 0] IJl1;mJi mkrn~:rgafll$tt .S. u~!(!JWii wullg;rnw mnl: fer:rncnl mannunl r{'S~~lllii'1lg In iIIlCid, production. whkh eanses the [lhmlll red ilulicElL r. 0 L+U:iI ml~ ffrurn pin kin" 'ellll~\r ((In:md ,on ri!JI!l.lltlif if I his I~lari' }_1'he left ImU r the agar P~!lIt .. v." moeulated wil h cm~!;!:,llh~se.nega[j\i'e stOll'by]{)~()~-d" Co u~li,lase'I1.e~,a&iU'e sHlphyhJoo~cl ~'I!HI gt:(lW Oil t.lleli1:cdirmn. bl,]l baeause the f.)rg.'lI1~~f'!Jl ,di)C$ not rerltil(ot '~U~lJillitol.. llirep.henO:~ [',ed ind lcatordees not ehangc in <color:. and I[hit: c(llml~ll;!. ~ml s :r· rO~l:ndlin,~ rm:diuLlil appear pmk. Bo' [olunies ,of 'l~pi!111«M'I"~ Iftl reu O!"I rn ann i to~ suit ilIg1IiI' f d se-u P I- Medl um u rr oumJJJ1:g colonies has turned ~le]I.ol>\G C'. (Jl~~;q te, Sh!J'IY, :.'{iliow colonles of SI~j)Ii~J,[o.t,v.c 'ij ml~'t'jIS O~I, i !Mtlli:lol ~aIt ,.I~<lr [close-u p I .

,5,·'13 OJtitcillliin !Screen plate. &reetIl:ng; agar plate .cOlO tniDli Ii~, b ji,g!ml oX.l1i,cilliin ~} 4'M NaChUippildHlelJlledl MlLleller,I:UiuJliQll, a~ar., used to delj(~<1 fl];I.'1thidJlj'l] (llIi1lfi oxadJJlnn;' re~iSb!~'li.!J~ m:imM (M"RSA~. Ol'!;;']dIJJn, Li> used b&;I1!I~e it is a better ~11d~r.:<Llm i!lII1jl:li.JJ~h; .. CiwwHlllhllJl:t occurred ;)InJJr L-,holrr .L1lc:uba'tilm .01:1 H"C frmn a sp l r ,;!1In ill culum ol' MRSA J shown m'u the 1,0,(1 and rhe lade f :IJ'!'O~ ttl or a tl'11,:,Ulnrillun-su!!oe:plibl!e'S_ "Uffl~ Q1I!h .. bottom.

5 .. 'I!4 Colol!! Lat nln:rp,llrilulogy 'Df Sf.aphykitGtH:;;,.. ~U;lll!: 9'Ap~)"Iiemlil ,g!ll! 5% :!!Sh:eaFi' bl!&Ddl ,0;981:". CoJ\onie~ oo:rel]]bifi It:l10i~ 0'£ ~O!.'l@llase''Il.egntM! :st"l'ph_jJ1lficoccl_

:; .. ''1::;' INC'VQ~~O~~ i6ll1~~em:lbIUbliIJ.t. us;ed to dliIe:ren1i;aj[~ S, .s.rjpf~llYU:t~lilt resi8fml~: nli~r.o~r::g~~lli:l'm till~: Un: rj~hl) [nm.1 o~hl:!£ spriss of (\~~UI.as~'l(;.,~tlVC staphylocrDm (.~ r.I!1.WJlNhlf; mreJ'7f!O'rnlmis~\I\n OJ! ~JlJ.c' 'rift) tlIlC!JiIil.IJ ~e:[tl!3 .in iL![[UIl:~ spec'I.nU?11:>.

St t

.. -..... .,' .. ","'" .-. '.: : ..

. ': rep,. ,oco,ccacea,e

D ~efnmi~.y =:::' ~ un~ _~el.~:ed gen~r~ ind,ude nine ~~~1~r~I.~I]~~luLter~~

In the cUnu;ul1cliJor,Htmj'" tiem OfLtf, A},I1W.(.'OCCUS, Bmero 0, US", Gt:mdia, Gtl')blc

caldlO, I.{oc.'[;O{."Ot'(.l~:S:, l~i:ll,l.r.m'ios~o[,! Ped.loca.CCI:IS'. and: Stl1el'Mcor, u~., The more commonly

iSClIHLed genera a:re lllustreted here, The streptococci. are c2llta.la~-e'"'negalti.ve'., grampositive., 0'. enid '0 occobacillary in. morphology. Ifl:rming pairs (mel' chains, ('0]0- nial morphology' and hemal sls on .. % sheep blood iaI~tiIT nrc' '. ry helpful chara - iter.istks used for preliminary- ]denti:lkaJ:lon. Hemolysis:is classlfled i3S alphabota, or gamma.

8-11 .A!!phal 1!I-:ma[ly.l. Clifi 5%, .heep bl'uud ag:sl" plate. lllp.rno Ilca'l'lO]ysUi is :in .in.iIiSIli1C~ zone of pn - U a llysi:s uf red hlood C'CUs (rUlC) r~using <II I!,':nrtlll ~u greenlshbrown dR:oimal:irrn ,af ~ [med~U'!11! iDnmtlclitiooly sUr!'(Du __ ml i ng ~ht!l culonr; - lrldans ~lri::pitoco\'~calctlklJlie.s(lllla )lneUI'l1I . t5l!Clil. ootnic!lllllt'"c a]p:htibIi\IIIlD~ytic,

16-'3, Del. beIl1QIVi&ii- on ~%, .h,eep blaod agar plate. ~~'ilh bitt a ,hml'lol) ,;1!':, there .i3 Cllllll1leole Iysls of R19C urronndingthc ml~m'y rhat cOIn madill' he seen ma 'litl· sco;p:l~aU~~.

6-2 eol'a.n, of iiiUphB-l'Iilliim<o!,lytlic ull'Q,pt:o~Dccl (;K'iio'l", MI("f"OSropic 'I.l'iew of .aIIpl:I<l-heml1~I~·8Is showing tlhe p;tTtially I emoIY"1.<2d lUll ilI_'Jl:it;'1.Ii<l~lil'l.y !;!,Il.1I;'O;lmdll'lllg, Ihe coiol1l ~

S-4 OQI'ony.' of' l_t.I-l!ieftllu¥liIC stre;plococcl IdOl. l\lic.msmllic viilf-W or b "ICl hel:rloIY!:lI:o:: shoiil'liiflg tlre dear. cnlnrless ~[llle around the strcJ.)tu;~"C8il colonie ii'll

hidJi die RiBC hsve lHLdm'l;rJIlE- mml!!C-tc dcstrucu n.

&.5 'Gil!li I'Hllftll thamal(¥BIs ,alill ,Ii~ B'li!Ieep Dl!lJ!IHt ,;g:~. -Gi\ll~~li!M I~ ill term u~illD d'erulte ~,';]d;: .or 11e:~1l0Iy$~1l:1i1!! me :sunouudhlg the coloales are W.l'~l .



'6 .. 7' Gt'Ia!t'i! •. Iatlnl ot &kiB!pibOCIC!oOe~ In t:!!tlMn, 'CU:!liUni!' ~x1~=-:Q'~~ GU'.o:!.m 8i,:ruin 9'[ ,;3, posiii,\I€ blond c.~~ntlrc i)rl)tl1 ~IDtJ!}:;;U'".t~~ ~g g1'am.iiJ'D!'1it~'Ir~ '~OCl.:i a[r.,~u!!l~'(] im cbalns, sm'pWt:t.JoOC:i 3lf\t:; normal !l1iCNblfli ,I ill the lCI[Jper [["~(Ii[".dQJ'Y tract ~1Jl!d. ths g~i!!tr,!)jJrl ffic:still1 al [rol:!.t'or thl!i reason, Gmm s1i:simoi ~l1Im1L.'l (rQI:J] 1(]'Il~tl sit~ ~ [lot .ilellXl1u1 lL1 diagUD:~ il'lg :IlrifecljmiJ.~ c"Ill!OO~by the ~ .. llm~[llc ~l.i'lJij)l<OCOOLi" such R'S S. ll1#JtIrtle'.ll a:nd S, rm~wmmllJ,L!'-

,1I;5;··8CaJGiIny" Of nD:nhemollfl~c stre:Fft,O~~(!;~ ~:-)f'l!l!i;!I',,, M:a(_~r9-$C'Il\I!l-~~aL~ there is ne ~pparen~ h;CJ.1101ynk ~It~ l~'I!!lY {Ir d.~eol()ml~(Ion produced b;y' titE) ('tllon~',

s",a Colulil ile&, 01i' ,gniHillR! A ':I!i,tre;pil:occl!!ccll am 15%. riliEiU!'!iiiP u:tilHid;agarr. Coloo1~e$ of. ~l"O!.Iip A steeptocom:1 Cf1:l5'i sheep bl cod ~g~~'H' are s;rrL2l1~ 11 CHI I!IUlrmUinled bli wid.t'! :rJUlI.4lli oU b~lllL hml ~@lJ,!:!iis,



~i-. A, CQ!'DnieB 1M 9.FC:UIIiP A ~ptoCDCC~ ·0 fill !I% .~eep bloodl agar (ItiJ:fUand :Stf'eptaC:OCCI.IB .electlve ·&Bal' ,(&S .. ~ (,....q,,' n prlmHfY sclocti\11e·l!>Olati n I (.ilium f[Jr g.OOIll!;l A s>trej}Wc c~!. Obs~i. -. tha L the ,oo~rmies III the &'SA ~IFe smaller 111~ C~! be cen 01] ~he plste o Itl'i.e d~hJl." Gnu;!!»' C, f, and G can ,[II::m be l::wlutelil fmm, the t~ppcrlf1.'1l!J5Ira~;:Q.ry 'll:'@.ct. ami II.h.~iJr cclol]ies 1C~n. be eon lused iiv'uni :Ii: [lJIlJ!P .A stl'iE."pl'ClroC_'CI 1 f ;% i1bcc:p blboi!J~ <I&8!r. i~ used. Inorulanon of i'I .I;~ooth~e I.Elt'(lim:n such 0)3 t!d&, (Jontruraung :s.uilffllnt'~Ew!{,l;f.mle (23.7" Ip..gtml), and fi'imethoprim. U,25 ,fA.g1wI)., iri!bfbits tile grffill~!I:IIJ{ ]]Q[~grup A bewh molyli" slrepll "ocCli uswell as ~I:a!ph}il Cl){llilli "'-'"iH.d.'fI!Jlls !3tt'etitoooccii ,afnd~r,iIIm'lle;~~1ilw ~[t.;:Llll. B, CuIanIU:I![:, 0' strGP-fdlG:'GGGUS: ~n~s '0., 5-% .h •• p, blood ,1I,gIU'i., CuluJ:lie~ .. re smalill mm ; Iram;luce:llt, and ~UIrit'{llJ.tu;£"'d by,[JJ '!i"fidl!: zone ,or beta hcmo~ ."Sl~.

6~'U] IPYR f:e.t. rnli!C' :l?re~tlce uJ i:1ll .arnl[llcpeptid ase ellzyrne that degrade Lite subSlntm-e FYR j~. ,yrr,· Hdon. I-fl~ I:Ulphthyltl~niuc: ,b,; <I lO"llJ!ill~li~' pr-esu~pt1vo te t forgr.mlp A Slrtp,t(H,:occi Ibcl~.bemotylir) ami ~.·I'(leocrn.s, .P.lw~.t'm·rn::!.,.1.~. <lml 'll'mdJu '1~.lll'h'l- r nnnh molYHr}. /Ii positi\'e tmU is It1Idlcated b a red color IIiU~.I: 1m dl"t rrllll! ,.

8·1'i11I1 IBi!;Iicttracin .u~.G.AtlibmtyIQ81 .. , r.il!3 haeltr<lcLI'lI ·.]sceptibnlil~i test is .an aite.rnal,L\'e Lo the P'YR 'iest ror the presumpttse id1mtHic<l~ioo or ,grou.p A b~l,a-henll(lJyt1t.') ::;:[;]repl!]C'{Ic~L i~ 0,04-U bacitracin dosk I:> f.!1~.ce{1 on all tnocuhrm of the utic~'oor:';i:iDlhm Win she p' blood ago. A'FI.'Il!X' overnight i [U:lI00:1 i 1111 !'II. 3 s,0C;, ;.~1Y ZO.l1(: r iDh~hil.i(JI'" is jn~m~· i,l["L1lttrd ff~ a .p~]~iti'I\B' 1iP.'::;!·, an ell he: fltlic.r'UIJ['Ji!..'l:m LSIIIl is pres:I:.O..m.p-· Hvc!y nd_ ltilied <IS ;·TOL.~Jl A 'neploc(]col b~f baC'itr~'l!:'ln.

8 ... 2 Dr.lml 8tuinCd;yag!inH~Ii8CI'V~C!n. ahm'l'i:.i:ug e',pithehal ce~is ~md gr:<'I!I1I:l·pooj.tJifl:'e {I©tti io [I'a.ir;:s. 5Ugg;e-Soti~'e ~r ~S'tl'~lP~orood X 1l50h OroliJlP B ~tr.epooooccL. S. agtdac,[ilU'. [;1:1:11 ,ooI0:J111.'i;l' ]h,e g;e:!lhotll_fiIIi<ll'Y U).ut ~f wmnc'l'!.. oo~lcmalUy c~]otll.,;lllig, illl~l ~TllrrL::C[I[lg the I1CiOJUI[C til ~~t.1J~!(it".Ynlnd thcro[ou is (he:! 1lJ'J.fI.j'eD[ cDlillisil! of 'I'I.eoCl.m:1l1 ~fi~ and rncnil"'lgilis. 'Micm~ s(,"Opi~~llIy:. Ibis species U!lLII~Il)' ,OITDrll jill. [l~li.b"51,. Hllllh.r,iwl'lI in t:futs nglJl1re-

& .. '14 HI;p;p !;IMrie hydmlya'ia ·teal. Grolll,1) J strepto~ooCii and :501n:c crd~occi call hydrolyze sodilJl.Jll il'tPp1.U''Otcr, fe8l:ltl:i'fiB: i!l.1. 'lb.oe Ib:nt'l:tilimil ID.f i!ihllCiIl.e all1d oodiruIiJ be:~oate.A s!1il:S~1l of I:hemicroorg;;1I1i:sm. N;irlCU~t:ed" £QJ.' 2: hears ~I :3 5,"'C l~ a. WwPlJltale s'!)lu:! iO~l ~m.d then lli(;; 1ndlc~lol·. rnnliY'"' drill .. is ,ftidd.cd. Dc,ull111talioil 0'] gty.cii"i:e. if it l!lprtseUJI. is deteeted by tfuJ.B! dBlJeilIlpnR'ilJJt oE H [JU11"ple, colbr t'l'!U:LhilJl 10 mIJ:RI ~~ (~,k![u; 'Oll ~}!\iJ' l~ll,. A r:JI!:g~ti\i'e Ife"c:tk~[! (~tdrr Qr~ t1~c: l~igh!J .!:eI]l~~n1i cOIO:r!(li38.

'6"'IS. CalanJ8s ,Id !DFaLlP' B S'liHtpmiC'OCcl ,iDnl 5%.lhaap 1~1Dad a_Ii" (XU)t- GOOt]P I H-epliocm=cal eelentes are larger druOln ,oitlhcr oot;;l,·hemob'Uc streptoeocct, b~.t ihe ,lwm,olytlLc: ~1):t'Je :5W'[loundlugtbe C:·(ilLoo.Il)' Is stllOl]er: 't;.1l1ll1P.i:UJ"~ ,:IJ:OU~ shw: of bL::mii'l fu,~nl![JIJI~l::; wilhm:b\m-~ [i,~~ OIl p, 3~.

&"1'5 CAMP if:es.t." A ]}Osbthre CA;,\fP' rest b: gro~D;1.l Ill, S!trep' ~Mocci, dcmoll_str,£litiiilgtb:c ,nr.rcj'~v1reild_;"shf[~ciI ,el'lihQ1l1(lcme:rnt t.lf bela. heillwty'$i;S '~hal OOClII[S\;'lh:eFi the tle'lTi:ol,YiiC hela.·tolilll I)wdu.cedby ft, r{lj~!re!j:l: ,tt~e: miCl:!OOrgaIDS<ID s~e.llJked h{)r.t:01!J.~ l~1ly ,000cr.l'OO~ tile :;h_eei;! bl.ooo agar ~JOIJte in '~h~:i1 pl\wl(il~ph)~d~ $YL16[~:tlc[uUy .viththe CAM.P fadO[ proWlll IP[{niJL.I.'lJud ~. Group 1:1, streptococt.:.I. (str'e.:iIIlredl [Ji8rpellrii'ctll ar l~o hh.c st!lplUylo. 'OOC:C-ilJ:i: but aot '[lUjrte to!!,iJC'htri~). 'Ihe CA_MJP' test, \~lb.tCIJ is named mr ~ts 1IIs~rs. CbrlslJ~ .. htt~l1S. and MUli1£b~GrS{lIJi. is an &~lf'ffI.t~Ii~f~ to hilppllrnt~ b~lltolysis.





18,.1:S OIt'a,ms,taJII'IiDf re.splt'Bm:ry .eQ:t'etlo<ns dlemD:lls:trarl:i Ilg grl'lJi[i,-DOiiiili'l;\e, ]!IIf1!(.':ot:~·:lhap¢d d lpLococ:ci sug&r:S!lil.e'e (lr 5'[.rep:~~~,,(,ijS ,~i1eW!lCmif!'f,

'8i-1tI7 A,'Co(gnf~ QiI' Slrali!!f~~!!J ~~Q. ni~ ~"' S% IIlhQGfj Ibh:iiod, u,SBr:. l'hcse JlOilJll!J; OO]@'uJi1til: arc m~d with oon]Il~,C'l£ oog",:s. :S[)IJfiIG'Ji1i,~lat mucoid and ~I:mul 1 mm,:i [If diiamelier. They are surraunded Ily ,:1 ~Ilie- of IlIlp'!a'ili 11 em oJ)' !ii:-;:" B'i CGt!;l'nias ar Sf!l"8pl~~'~'Us :P!it~!Il!i~,ffi~! !Cl!n ,~:% ~~eep' ~Qg~ ;jj,BlliliF. 'l'hmlt> sligb~I')' 'ulIBcr coleraes display eire iI.'l~~]firnl Im.l'Cnl,~HtiH CillJs,c,-'ti, by the eas:lily lui.lIllC.oo tiutifJolys1s Il?in)i~'i:l1 ~Jf S. ~Mr.JlllJ\jj!IMe. C, CDI'anl'es IDfSfr.~pftU:;DtG~'pn;euntanhl!l!l' Ion 5% lIih!lll!l:p Ib'l!CIacil ,Digall'!.. r:lm:~a!';i.fln3nY., .)" pl'fi'lm!o,nl'(!4!' ool!!ln~eg: n~."J¥ _';:1:1l1:f!ly fll<lIitcn out <J "lh.cy ,"!g.e.

&"''II'B .lIi::GilonHHi ,or ,s~tQ~gl!;l\~I~!!!I;,pn!llll!lmQ,nfae a:n ah!l;!lQgkde ggar:.1I.·tedlIJI'l1, WEIS lncubated ill COol' resuhlng in [:ai:l:ly JHtgEl aones o.J ~lpba henwn, 1 "B. 'Coionie'li of Slll'&pla~QU8 pn:eumCllnlae a'n ,chocolBle 89ftI'" (C "_-UP'_ CQ.l.Qru~~ are 'qui~e Ilat UI] chomlate ,'~~illi.


'6,0,18' apiGeHln suslDapl:lb!llllq' biet,., Colonies 0,[ Slr~o.!'tI{·[Wi ~Ilt'mmmliie. L'l1Ce iJ1~'l;blted by rb.c ffDl'01tl'!!lC'I'lIbic aptochil1 'el'hylhJ'ck!llc~lplr€il~e h}'dndtlortdc} ,~Jj]lHlI]ed m the paper dbd:: appl1M t(l 'Ihe surfaee of all iaoculared 5% ,s.~eli bload ~l~'lr plate. A zone or greater than or 6qU8~ to, 14 !II])], i diAmeter Is [n'Cswnplltre i.dientificalitm fm- ~'lI'i.'P!(J('f I~t"'.lm [Ull'I'I'JJ1111!?-"CI~Ol'l;~or inbibjtim~ is romi, lel1~, wllh l,'.ilid"mssl.r{.v-, toecco, ZI;,l[];~'iS .of less than .IIJrmn in diameter arc qUC!:itiO:I:I~ albic and slu!Ulld be oonikmt:cl wjtl:l ~IIL: 'bile :suluWliity I:l'~.

-2.0' A" Bilel !ll!D~ubUity ~:. :"t f'I'IJ(OCll.f(JJS J'II Jul'toilI tf" cuu'o[l'ii!'l:S \,,.ill dissoll<e Of I 'S'~ wltnln 50 f nures a:t 3 5~(: in I:ibe ptcWDClli of :2 t~ sodium deSl"L'Q'CnQlate. A.'I p. f the f13ilg 'tit ls ap(llilXl direclly to the colonie wW" ~ dlssolve fpJilfR. Cln tile rf9'llU Ii the Ol:gm~;!iln!!'l S~ltJ'JtOClJj_"d·ftjS pnt'.wuOl'd~f:'. (uhllli~ on the blond. ;1g;U plate onthe I ft. ';:]1 (jI oliCr!aycdwulh I.)tle;r>emaill.lnllii;!l.el.lndic~ltiU1;g that '!hc ioolO!itl:e 1$ not SJrfliw,nxt'U:l: pJrl'lT1t[[][llnf'. B, 10 lie, 1S~I~bill_ te.1 ~clG_-up)o S't''('lJ.t~Ct."'(.'jjli ,i\,1!IE':t[rrwJliull! mlQoies!t Vl! dissoh1!&d I [1 th~ !>lIE! udded W the [ll:u:e su rface .. ~fter' ac b!il~ has ('\Iifp(ll'a~ed; o.oty the sene or hC!llo1y'Sis rerna i [IS. as ~hCi~\' m ~l reo

0 .. 2'1 AIiI,llm'I~F!Qbl:al~ UILi!'II~p'tlibmty wllltilng v1i Sf#"eph:!>Qi:!G'C,t.I. ,p,nlililiiUmiXiIlilil.. S"~lrtP'loJ:\C()~,HI:i: ""1!~ll!ma .. 1i'fJi? C~1II1 be ~e:oitedl [(II; IJrel1iit~i!ilin :re;!ij:r;hmcel!l~1l11:~ alJ) unit pen ic.:il)in diisk <lI ~. 1Jl~, ~[lomcil~in disk {)rq~ ,L[IJiMi~I~1 with I .. he Eo wM {M ~jool&k. Sokm, SWCi:rel1~, (lOOlJilllting elf :UJ pl,:::!!l1Uc ~lr~I1' hlil[[lt~1f'I fi t~rI wWll Irncr.e~sililg concentrarloas of (he O!I tHI bl(l(Uc nnl~l'!fl::; case, .!J~.Tljcmirtt FoUowtng iI}liIb'fn~f!;b:l ilrIcllbalinlil .. lt..e prlirlll :tlI'~ l,v,niich the IBUl.pli[:al :limn!! o["iutJibti:HuL1 or ~it':t;1n. aU 'Lllfl ~ool."jtei t1f1"Cil'SlIl:cl1'! 1hflQIl amrlit~ti'\te :SCi] ~1Il 0111 th~ strip IS i lIi!:erpreted asrhe rnlr!lmOJ'Illllllhlbltl)f"Y CflotM;'e!1l.tI":~il1on nuilC)I,

8-.23 DptactUiiii SIlISCU ptlblllty'le!iilt: pe.r,[uttn,IRdol:'i OJ ~les of vur.irl1ll1S streprccneei, C{lI~lII~s df~'il['ii[j;jri!S sll['"tllJilo~ rocC'~ are res:lsl:<llnl to (I!ptO(; ni[l. (li5Iing!.]islli ['II!; ~l1ese camifIiOuh'crJooi.1i'!tcrcd Ms::ml:sms (torn .S'~I:llPI~ft!"rrS ,j~!~el.~"!(m!'I'1I!' (;soonll il]] :W.igure 6·19,.

B-,22 Gram ,slHliii' 1m" Hi ¥iIIF-IJCI\i!lIi19i ,sti:reptGcooDUll!!!! :l!Ipl!lol!_ ~n Ibloodl DUll,lUll:'t!!Il IDt'oth ([I{'\'I:2l!SO). l'her>erili'e .llll,lbn.erQ1;J s $pecie~ or the ifil'id !ln~ sl't;etJt()COCC! g)t'IDU-J." whic~ are Illftm<11 il)'luc'I)S:<l1 i~kr(ib1ot!1l ~rJi mammals" OI1.I!fill. f£'I'.' :D:eql:lire hhrm l;j£i(:<tJi~.ofl to' lh~ ~.tx>des lee wi. s uch 2I:S S. Ilf!'f'i~. ~\1lkrQ~t;!pkally; tht:y l!iI::iutlltv a pPf;tarin !jJ.[g~ ,[brain:;, '1';:15' ~el;;ja Uy when fe(l!1'1'ered f[\[)!11.1 Ill! blond ,t:>III~urre bt'Uth nl~ditJ!m,

B,,24 Idan'liiifl!Ga!IDmI ,d' '¥ilriidlBll!I&' .&cb'apmiCoccl' w~th!l;;lOFliU!.ntlDn811 ~1'DShliilm'I'oaE lI"Oa.eil!l'onlll,,, El'6lllli· tiveirlell~i~i~~ti!Dn I['~qui res ~\rend substrates ~im::illl{l.lng Ibi~~ esculla, ;:]I"giinlne de<:<1fb(l}!yli!l~, ,td% N .. CI, II-'lIICtil'JI3e, rTrlI111Jil1itni. rlllml:li~, sortlito[ll. aeabtnose, mulln. sucrose. and ,~cullu (Sl'itillWh. htlrt'). Qnh!ilt d:iiI.irl.lC~t::.i'.iiillt!s ~ILfCb <IS ~roii:l;rt, V(t<J_(:itfwslhllle:t re.l:lttim1i, Cbije ,ul100llllbiltly, {lpi!J~~ i til. resislil:rl(}[l:, !itld &"lI.~~IJjre to gl'J!J\!.' ,11'1 ('II~C 11IIilrl ~,5~C, wi~11 h~lp Ijil:l 'I:ilantify thecltnIC!l~Il! :5!,glW Il'k"lI_n;[ l;solii!iJl'e~,

,8·2." CDI\!!;I!:ni!:I!!I~ C!II:!\" PeG'f~~"a' :GPp. gn,5% Mhe!UiP bl:!!iiIodagal:":' f'i'dtot.·~"('!i!iO spp,r~IIIlh~c ""tr:ii~m:ll;) streptncnccl ~~! their c~lol1kl1 !llofiPholog~!.lrlJIt liI1iffi'V1il;op~(;aib' th~' I:lppelliT'~:S palm's, telrndil"l,ancl eli I :':1i!i'Ji;, '1lrf~1~:lIlliiinL}' be ~li:sI±!'Igulshed Imm, ether ~ilpifua,;'~Clnol!i'tlc moct'f)o:rg:m~lsms by Itbdr ~~L5t;lfl)Oe to 'I.I~I'l'@!llYC~fl (~f) J-!l,,[! d~sk) tsec FagUl'lJ 6.26) and grt)lwml1 ~I'D ,6. '5'~~ ~~Cl b:"r(l~h, They ~lroC also bule e$('uhll.~:tive.. but fl',[l.-r.tCg~liti,I.i'C"

,a-27 NU!iiiIii,u:itlCllytic l!Colonil@. onl l!ll%. ,.eep IbilDOd ,HgiHf" platill" in lheffouMi ly :.S;[l1':J~'lOro,crm:t:'f!ll. the f!Ori~l~nolytlico!:g,am~LllS: i[l'l:'~tl<le the Ell [eroOO('t~~E 5 '[l'j,'i , , semo :'ltm~r.I!s. fif\l"irtdilj]~ !>t'['~ptQ~ooct" I..ilCill'lI: oceus spp., ;aVId 16!1IC!Jlllt.~.tj1[' :SP[l,

' ... 2. Va!l1CQHlJ!C'~H ,.",c8;plib'i1iiq. il:liilliot. ftqN~['(N~ SPj). are dis[ulj;~tljl;;li!!ad from uli1eJ,- al"hil.11I1mlOtylic' m~11- IS['I1d~'llle:1 res l;SlaJJIOI:: :~1I \I!U'lC'l]ruyr.l~. ThITe ts no z(]n~ IJIf l:lTIih~Mnl(ll1l S!UIi,It(ILUH.!J fl,bl tll~ \;·aI:IC(]T.n~e:iin dl!t1iik il~;ni::ed 011 :3.11 IIl.ocuh!m~ f}f rt.rJ'r.llt.l.rd'Wi spp. I~ riOflr side of 1)lflt!f:~ c:tl!nparr.(l to 01 :ilWl.e of .i [1.1~.illbili(nl ~lI!rml,]llidill~!; the v~r]dans ~rep~OCOCC'll:i: !leJ~ sM.~· oj pj{il1e),

8 .. 2B 11!I1.lal rQACllillrtn, alam'. Srl;(!!lfliKi!]jX'!I.~ b~~!!i\i and the !':rl:n!l;':r(l!c.:~~i, ~.ti!l ~mw ln the p!Jeseil1tle 1~lr' 40{,!il hJ:!~ ,1I}d '~!lI~ hy· dkllJ]Y:Ile est::t!lin brl escelen u, f);C(11~tnn ;jm:l r~rr!,[; di1r~lle rbrm a black C!)11'IP.!~~ h~ the '1::}'l!I!', Will ell C'otIi:flw S 40% Ol<':!;lillt 1\ 11G,gaJth'{! :sJ<.111~ reraalns ra 11 colored (lliIJ' (lDla-1' charl~) ,ootI'I;pared to bl~oc:k culored shlr~t ~ pu~ilhe shown h~),

e.... Cok:m1B1!Ii g:l! ,EPd~~.n' -pp. an 5'% .h.- -p Ib'loadl 8gal' plaite. The [oklJ!I~e8 are r;;Ji&erll, wllal;ie·tu gruy.white. r.:tImgc frC)nl 0.5 ttl 1.5 J!ITI1a. in s!~e,. ~md are l!I8ul:lll), JumhemotyOl:!;:_

18..:3'1 00:1'011'1 •• lof Leuco.'llDSloiD SARi. !lim 5% ..... " p blood as'a!!"' Ipraia. Ill.ii,u8JlI.y I lite c I rue resembl'e!l'l1!le'n'lctlCl.'llS :t;pp. as desC[ib~1 in IW~~ure 0-29, U.POll Wlllil1lued IlllltuibElliurl t48 kl '1 ~ h()IJ[~~. " weak nlJ'lha h -'ill Iys'rs tieveloll . IGclll:!nlll~', these .Il1fLCfQOrg<!il'llsU'l£ can ~e distinguish&! mom fm,(fnoo ,I,IS :iiPp. by Ihcil" rcsilOtF.IJn1;'e 1;0 ,~,.ttu:omj'ci[j and by ,gfll£:f'Otimllion f[\illmghli:::t1!s~h:11 .. ':t:am')",'"tCrul r;"ms oc..otb (Difco I..uln11i rles, l)etrnil, ~lih.l.

8 .. 30 IRlfe t!I'.CiUliln iii.ant and '8.5'% N'sCI bralh. The blle c:si,;uJiu.siafllll. (f;!!il) jl;,black ([}~IU ... , '1. il1:dkating, ILb.a~ ; tlteilllic[·oC1rg;mism call! grow m the pro:M3ill~ of Ibi!,· and h 1·dJ:.oll":Ii~ esculin, Growtil.ln " bmnlll Co.ll!..t-~inirig a salt ,concclllratilJD of .• 5% [,ltI~f.o,o ,-10110 ir~di ·',m'foo ~y·lurbid;t· and ,I change in I h e indicator lrom pill" tn yellm,\' foT!ill\\lln g UYNU:llightt ulDculmtlml, Tille reaeuuns shrn~l1n here IC(lnfirm,lllc Idel1lm OIL! n or liri~,~j:otl1tt'W~ ~PJ;l, A IlCl"-liiJve :I1YR 'lcst. [Flgure' Il~ I U j mail lIibio 1m LI,SGd ill mn:liilli'm the Identiilr;l[n:ion of su' ~ pc -etl j!jMi!fin;at-ell~ !lp;p.. ol~i',l'lllerooooc,jfui .~l;1:'Ol]p IJ !Sl[ll.>p1:ot'UL'C1 (6', 001.'[!1) ~n'e unable to gOOl\f ~Ll tlra presence of 6.S%.

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CHAPTER' 7 Aerobic Gram-Positive Bacilli'

d'A f

ana r cunomyces spp.

ommon genera of aerobic :gr~m~m>ositi\i'e aa un include B dlirls, Cory.ne#JfJltrril.Jm. ErJ~sir} ,jaO, Fix, Gl.wdnpreJ'la I gram-variable I, Li:'Il,,'Lob:(!:cml~s, and tis-terr.a.

R!Jite clinical isolates include species of iln:{lIlOI1ac[erJW~I. Oersko1.'ia" and R1wdacac us. Also included in this sect DO~j are '{\ro{,lu'dil1l and Str:eP·~0\!119\('e3.gene:n~ 1l'lrJnlaining tiilu~ mentous, llilrancning, gram-positive bacilli.

Prelimlnary dlffe ,I ntlatlon among these .i:!en ora :is Iba'~d on mleroscople tncrpholag}.7 and the catalas reaction. R~tmus spp" Corynd'art!."riwn s:pp .. ("JsteriCi, Oer.'SkrJII"I,{l1" and lUrtuJ{)j~(} 'us spp. ar:e caralase-poslth e. Of these, Ba.ci"lu~ spp is the only spore former, WWre examples of the microscopic morphologies of the gram-po itive ba ill D arepresented in the Gram stain section.

'7 .. ;1 Cram ,StBlnl of' ,Dacli'ltUs spp. 'Large, grompC1silhre imciin wijj] squ<':Jr€d-nff! ends mensnrtng 1],8 X {,; iJJru. ocmfrill g: s:1n~1 a nd 111 chatns (x 1150), 'ftiey m~iiy appear ~ra[ll=\1al:'ia'IlJe. Some slP~ics cl(l'tnQii:'itj['EIJh~ I3llido50()Ql'~, wh~ch :m fly' 'be T(JICn~cd ('otlltlfi'l Uy cr ter .oall!J:U'I!1Iy afufig the bl!.'lcillL The mle:r~scoplt :lll;orpll~l,Gg}r :tUIl}' resemble til at o:f C!o:nrid:" iilm ":pp.

7'-,~ CU:JvnWuu, g" BaeIJ'.-.,., ~pp\. all S·% sho p bl!!i!ud iiliga!J'. Coim]i~ of Hil ,m'llS sullI;" .are smooth. round, <tnd IJ.I touuth .. tI, by ,I :i:mll~ of bet a J1en:nl~Y1'.lis •. 'tlllml i:":;; ~311"! 0111 I) appear wtll,;b ~11~ dry wiJlh :-;;btlggjf ooges.




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7-3 ICalon', G1' B!!iIell'hI'-, ,- pp. on s,~, "h..!II!iF!i b Dad : p~ Some peetes of B,adflus are IlQ!lhemol-tic wilh m mid ;~IIl(l sp'r~_dbng colonies resemblmg ~be fru~rohm;rl1:ia~'ed" ~M t':sIl';r,t{lwi!~IIIi~ !fP'p. '~ionie;s ,~f Hti,mr.r.s mllflrads, the m,.uor huwa:u :~alhogeR. are II.I:!: lal~1 non! m olyElc. TbLS 6grut:~J1tJor,,\'s ,fJrn envtronm .nLalSadJJus sp, that rurelv causes hum .. ul iU~Bd inns,

'7'~ IDiiI!I&tlIn hydl9O'l"i8 ts,t, Many Mm:illW" spp, ser.LJt'lEePJf'(]'teol:~rtiCeJ:lzymes lbart eall1r L'iiyGlroi}r2:e gelatin. ,I" I.he Plr;:cood 'slwwn in, lhl~ 6;;1;,.1.00" t'!1!prnlcd, undevelnped x"r,~ry rum is ,,_sed s 't~ ilic ubstrate fur d 't.eotion Q.L Bej.a.Uml~e ac[lvit~ :lHl'ydro~Y'SlS de;,tro'j."S the ~ IIl1 's gelat.ln 0 H Ling, I -·~~viil g Uillljl the dear p~'!OI~r~phi!; film 'i'!ll~c~c' the st rip WI.I;!iul:]]lJ:mrscd ill ~he 'i;lTg2l11j;:;1l1i 8IJSpeIl ion.

'71 .. 4 'Ot'G'Wlth 0'1 .a~Ulgiil ~p;p. an iI!I:gB 'W:!DJk: B,91(!llltl. lileadioD an egg yolk agar lis urt~ ur the char.ade-r.iisl:b:s nsed to id.entif}' ~trj'j.l!'l~ :<;PI: BilCilltlli l't'MUS' Is one of man pc ret :0::: thai p:rOOU!l."c lim en~Ie IcciltbtblOSC, demonstrated b a sone (lJ opadt 1(. hlt~s;h (\oll~,r ill the agar) ,e;o:;t\:ildiJ1:g <!I'I'!i'H}' !:rom the ~ac!critd gJi:owl.h, us shown OJ!l, theldl. Lip a e adi1rjt~rc<'"Jn.:11 ~1 b . detC:Cledi 0IJi eg;.~ yol"k;l[gru:. ,!It.!tl1ti1ll~lr'il~ed, by' 0111 o~bv looking sheen Oil thesurface of the baC~eTTIal~!r~w~h blJlllOlI] ~('ndiT.lg OJ1il the "Sat', as ~howl1 on th.eJi!!,hL

7',·a Colanlill: i 01 L"faRa mcrn',DGytagane4l1,oft

%.111 __ ;1;1' brio" qaf'. C(i!QtI.tfi:S oK Lw.~.rI!1 J'IQflGf!,Jlll!Jell.:'S 31L'!Il SiI'(D,':1l1i lie than 1 mtLii jill dj;:J'[lIlf.'ltm·)I, smooth, l!:' rC-£tTItar, ~lrId tr:.u]si'!J.c n~. The e h,)nl~es m-e ·uf'roo_nded by a \~ry charactertsnc fHIIt'"['O'" sene of beta h~mol'y$1s, On :5% sheep blood agar H[iU Jn~l!il be oonrll!ledi u,rllh ~oup B oota.hclIloijlUc Slf<ll:plOi;:~tL DiITcrenli3t Il'lg char~C'Dt'rist1C"s iil~d de U:s:rtrlrl S DH.FWlV 7.oftDe or bet a i'lcmoty is, posiHve catalase p.roduc::LicD, 3~'1d growl h on biJc and b,}'lirol~'sis (If es:cub. ]3.ullJi :>pel.: les hydroly;:c hi()p:urdlt'(,

7~7 EQt;;!II1Iin, hydl'alyMjii! te~t. MI;I1i>1 Sb'aU.liS of l.t~~r.rj'a mvn~r['}J~lJ, 1.,'i'~c.'I' ::I'll hyd:rolY;-lB !ll • ·111in lio less than 1 hams. This: pl:Ji!!JIIJtl!1.':<)ph !>ll~Jo'l."!'S r,t'1J~H ive reactions fO:It bile esculm f lil1M" orr. .klt} and ilDppl1nl!t~ rI0'n:). These rests are d~5Cr:ibed in l'ii~IlIJre 6~·1,4 0:11 p, '!l0 oltC'ldi Rpre b-,l8. on fl. 44,

:7 .. 9 Cvlluni;g~ 'g,I' c.. 1.''''~':ll1m' ,on S~, .beep, big" ,1l91i1l1l'i., Oolmlies are smooth and ·whms;11 rollowing overnight ~n,,<ub!:lLion bl 5% to If,' 0°1_ TIlLSl sp 'Ie!! com eause ~iflJ-lhl"tm Bill! sepsis,

7 .. a Bram, __ ili1ll ,of CGI]i"".abac:f~m :_\pp i ,~ 121!1Dt .. Smwl ~ram-p sHive Imclili aPPooDlllgl'll p.ilis .. des. V~ and L f:orms reScitlbilll:!!!l, Chl!lle~e leuers, tlil:ut>1 or the

'DrY,I'li'lucl.4!J""ilIm spp. is!ll,.IIl,,,J inthe clinical laboratory are n:!1JtlU'Ifll fl\, ra (b.f the ,~ki[l and I1WOOIll.:l> tL'I~'Ul~r.rm:les, ~t is filimCulr ro OJiUe-renfJ.t!w· 'II~!.:· II I'IPrl~,In,(lf:l .r1I iI: gpecie!> rrom. tile pa.th,,· g,enlc,.. . €!rpllli'lffim! ,all~d C. J,j>I~,durf!, j)J1 the basis ef UUl Gram stmo.

'7-1:0 CQII;[lIIIIr... Gil c., ,diRhUi~ii' 'OIl &% •. !iteep bien:.. ,.SUr. fDI"Y,Plil'!llIf:t!l!r1l;1m' ~'jIJ11~I~erjml m'w well mil. 5% sheep I.illi'lJd tlg<lT .. !,!;;'ht~i8h., apaqueooloute:s.

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• • 7 .. 11 Cuilull!iglli!QiI! G!. dN:;;lfM:/u,tri'i!l~ un lii'nliii!!falu apr.. A ~~Bt:l:hlill rn,~dillJ]j] ~l1oL.did Ilc used. illoJl~ ~'I!ith :shef;p ~Jlooo ago:r \Yl'ncne'!i\~j" ,C(.l!'ynl!'~~l.:tC'niJm,aj~imfrj\:lr' is suspected, as 'ii'lrclJ, usto difTerenUHw C~ d{vllti!el'i~J1 rl'!Orl] Ofi"l"er oor~mdb,'3,(. ~ef!21. o.n ~h.e' ~Iedh<e m,~dilu:m. cy:)~im,e tellurite agar or potas£ilnll. ~,"UJuritc {l'wS(]<de] atlM. the colonies ilf C""!1IleMLerl;r.rnl diplli f.lE!"rirJ~' ~m;\'t! <L !,lulrIlJ],eli:l.I.I,l.!:<'Iy-l:!li:l,t.;k. ,~ppe<KaJm:e.

',...13: 'C ... :luJilielilll, ""if CV;!'J'nl!l!lba,.:;::hrri~Q1 8RP.. :5o~'D'le ('!l\rnI'JIl!:rm:icr!,[U,rI !llJp. (cflml t':lllil!t:d cl i,p.l:Ill:mrui~iS} il,ppem'iiS IImhoJTIt!!I.;tk"!ll:[;~.\'!o:·hl~ill, Y;[I1I11III Wltmi!!."S 8'1TiI(I can !.l!lslly be ccmrll:l:';crl tvbin cCi~finEr.!',l nf !ltre[li:Ci!coC:~I, e',~[leci;i~ll:y wnel1 100- ISlled. rrona. 'tl:lc ro~pir,f~I.ory U\!]Cit. Somt,e ~c\LCS appenr whll1: illld ~paqtil..e ~e::lem.t~lil'lg 00Qg~ I ~-~!I;!.:l!:mitr'le :naphyJooOO::i.

7 .. 12 l.<QlltrItI .. IRilothylBliIlB 1:1(1110 IIt8tln" C. .dipldlr..ffiFNJl\' demonstrete IDl~Jlcbm:tl1:BtiI;: gr:ll nules when stained ~\!ith e~ 1:h~I"LO()rllk!.' m.C'Lhylc.-!~ ~Il,r)i S'l £II i~ (ill' N~iSiiCr' :5t9i!il.l'b~ ~tiJj,[l ls baSI: p('~rf~d on OO~O'IO lrzs, g rM,t'.1:I ,0[11 a .~mtt apjf :sltrn'fl.. M..eI~.!;c-hr(l1ru<!i1i<: de[)08its are redidisJ:i 1lm:ple in 1oe(j].er m~I:hijiJ· ena blue :li1LaJJJiI.

'iii, ' .... ,


7 .. ". g~<Ililml .b::!il" Q:!' ~~fg!!!c:!Jr~ .~pp" ~'" ",aul., na1$~~,,(jUQnif;_ [L'1'rl\!il~ad}~wl $,~Fl. appear as ~O:r1I~; nQ:mI.·· f.lprlr.e-rorm.i~~~W<.l[ll1!-posi.fijli.le biM:ilh, Tfuey [!J:'W .0 ~f)[le .. r ~~ short uml.~kllJm{i:rph~c ,tiocrobarUn. ·WIU"lll.ltoo.y a:p~ar 1UIffi'~smpically ~~'L"ool:l[Ilmdlli, 'lillf:¥ can be nlJislti~]]tilijed as:Sllf!!.WLO. eeeel, la~~(i;"aoC!lli are ~(Ii]'llli<'1Illh)rn o,fthe l[:!ppero !:,CSJIir~JQl1' 'tract, Inrestlnal 'rrrac~, and ,\l~g[DiI.8i. elthcugh they Qlllen :~BiI to grow [I}, t~!ililliil'e,

7'-"6 catonls •. of ILaDllabaD.Nlu;II! 'liIP,p. C!:'l!o:n.i:c.'> alf-.e ~I ~h .. ·lu:~1:rl~y(jc rC3emMingV!lrld . ans s:we:[,Dloorn~! and UtJ}$i~ljIJ1i'1; .. It mill,¥ ~ ~.t~es:SW]!' ~o dliJ£Er.~Dl:iilte t!'lese genera i:tepMd:ing ,OJ] the .Sf,;1'eCirme!l scarce, J11 mClM C"a~~.oom l(!Icwl:ioaclJ.ll '[uu~ !11reph:Ii:;{!!:;d ,~ [WJTJlIml, 110JT'"d amJ. liD i:llilJt reguire iilenli[ical'i:r;ui. They ::r:re V:S!JI;a,~1y re81~t(I~1t ~fI\I,mlQOIl'!)'Cjp, Growfb !XI] 1'S] ~g.1['" IN III dlffc'I1Ci1l_ ate: E'r!1~ilWJj)!hrlx [rom laetot.r~~cilli ,'1",'11 lcha re unehle I:u g!rir:Jw.

7,·"8; Cv:1Ql'deB ot' B-y5:'pul.allnf"hf ,mUJll·j'iilI~fhi~., ('olem C are Slili .Olil:t, nnall !JesSi Uh:a:fl 1,n 10m in dlaOl:eten and lriifl5, iilillli1l. A sreenish CO[[)f 3r1df(lf C!Jl/hcrnol.J,'lIlis !:liP]1'e~il as lIlm Cl)b:li~ ~g~.

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- ..



7-10 arDm .t~11!\I at S'ali'l'dBtOif.\' _'/~ 'IIIafllnatiii 1(;>111125Q:t. Gr:in~iltTn'il.l«!1jiI1Jmlrf!> is a SL11~I]. gl'a:mi~tjvc fa IWdlll't-\i,U"iable rmll..[ike' I icnm:rgalllkllill that an be I, 0la1ml frnm the gilllilcm:-ul]H[Y tract of humans, lbfu uorgaIlJsm liiI"dS at eue lime i:mlcJ udeclJ in ~:h e ~~!'!Iera (!cJo,r:Y'rrt'JJ'1lt·~!l'ri.Mm and! Hu.t"llClfJ'!rii!!: .

7'-1!8 CulonllBlII, af GiBND ,f"Sll.iI 'lli'~tr#M'm' QII! e'hacafatu agap. Miler J 4 heurs ar In f,HllIo";]t lon, colnul ~ !]if Grm'nudrfr ~'tI:!)IjJ Ildis Ol,ppea_r illl tm~', p.~n~dlfll C{I~C)11 r:~ m~ checol ate ag~ll: h;olOHtiil)lI1 of tho iIlliicrl.llor~allb;lll is Illlnsp~cilkfor. ! he diO\gn s~ of Or tcri II vagi 11 os . slno • 1lL."l many a. ... - rJ% IJf 'l-vorru::n arc ('~1!JI'i.iz.Etl, alilllfi'llpl.~)mjJHI:[lJlly wil~'h G. ~m!1!!Jl'lJ':I"

'7-" 7' .E,.,.,~mi'N.e r'h~i!j'IVfl1ltftmlilii!l' an 'llrIph", .ll!J8r Rift (TSilt ~gar. Er!,4s111doUllix rIm, iup_llI'tilia,· demolllm!Lfres :JHlS, pl"Qd~~ciW,1.l along the slab line in ~TS] agar ~~allt '~'I'ithlrl <!Iltl h(mrs of inculnlti[Jtl at 3S'C. 'l'bls Is II ilr],ptul feature lbal separases this or,gal1i'>ln rlf,om [lith ' grmn-pusilivc 1 aclllt .

7'·20 Hipp ..... 't Ihydra' ,. .••• '~' iIIt. (i. 1W1gi;j ~is h!!,~ drolyl':cs :;OO_itlm h i p.p1J.llale. fbis rnicrou:rgau.i!ii.rn. by'rilroly/.,e!l 1 % ~q]ll(:tH.lS !>t!dDtJ:m tt:~pPUlrai~:erollow'il:lg a J~hoLU im:uib.aHrlll. at 35<1(' as deseeibed in r: i,gi.U:1: 6 -14 on p.4:0. _A.. ,lJOs:ltl\!e [ICi'lCtion isthe clevl!]Qpm(:tltoJ ~1,pW'plgt'IJI~Jr \~rithj!1 UhlllkIll.U.·il rutel the addIDtian I) r .Illl1 h,ydrin. /\ .nE:~ .. 1 i~rl:l fel'lJ{;i 100 .is l-'Ohnt· " 1:1.


7>-.~1 ,A~ C!!);U):nfes ,m' .AA:<aDGumDl'lumi ftOvmC!'l)'tlC;:mmI' [":oft}1 8n;dSI~pIk:!Cla~Ui!l PYD"' gena,:!II! t~""l1IM]. Cc~oJ1ies oW AOI"l"t1.lm~r:rr.J.erjiml hl'mi!D.~li('!!m are t:'l!1iJ' ·(0.1 lUI 0.:5 m:rnin t:li;::l.ln~e:r),!l~a.hen'l(l;lytle .<JJter 48 hours, ,'311d C3.l'l 00 cQl1ifiused Wlli1 streptocoed wh.cn i:luUttf<-i) fmJlI Oil 1:hT:(lflt l:1]1I'lre.!. As s'f'lOteVI1 here 0I1l the tG.rt, "ill: colcnies :11f'C' :t'!IQ.lhcrnoOlyUc <wUtCf o~r!1ii;ght inCi.iJb.aLiO:nil. comparedwitls Ihe :S1~!'(I!:IJg bela !1cmo1j'£ls of :gr{lIl]p.1.\ :5l:Iiep~moct.~. AJi!)llIU(lOOt:~ uriM.m ~,\i\iiSPi:'cvwtli:slyiiichJJ:lro ill the genus C~I'IIr!~~trc[e!'i!j!lI. but .It is ca:t;uil<lse:-i:l.egdltt~re. t'i.!ikmooop:i'c:JUy:. l(h£sf."gl·~m·poSiHv~ bacilli [l·semlb~e:CfJ.rymfl!jlc;!crj:l!ft! ~£IP, D. Colonies of Arrntl(lb.rlCtI1iYliuJllli~mCllfJ~!·nj~"I'~ ~~~l'l) ru:W St~n(Jl'~"fW; .i;iJjO~lmel'l HNlUI'JJ'rr) h;:~(!Ise-yp ~!l..cr o'Wmlgorl; Inctd't<Hlon).

'7''''2:2 (h:~I!an'WB CI.t' AI:t.Q~~~ ,''", I~%~~p IblDDd"gai~ RJ~rnk~!.u i~}C:~iH ed wltbJn ~!a:e "nocM'd:i:oUrJmls" gJ'il'Lip, RbodfliIi'f.!Ci'Wil ~1jl~~J \,ms also !.)f(f\1lously lnclli(l~ ill the genus CoryIH'~QC~fil'!.W!l as :well as iUh~ g,~Mrlll~~ MUrniJll'J:. terllJ m. The: m[c["(lJoc~n!lilm :gr(lw9\w~1 on SULrep1 blood Hg1If, and wlm'lll~ can rasemble Klf;~£it'llrl. but it d'oesl.l01. gWJ,ii' [nll MaOC'cmru,'y ¥i:. ('olo:F.i]~ ll:sii~llIy clemo!, .<1 ptl~k pigaMCTIli: <lr· 1.<2rllwl!'Jllged lmt'!ulb:iliI.tm:l.

'7-:2. 'Cili!IJlio_U gi' (g~oria ~~ on i51'A!~IIII!P ~~I a~:r. 1~r;sk_Dli!U!I are mOl~l(l gram .. :poM'i!~.~ filaITtbellilS: ma1\ fmg-IItIlDt mt~ ~. fmm5. Tbismi~'g;;lnis.!TI canbe p,resump~we~y 1den4ified ~l tts .micMOOpit ,illor..liIbol~ID\ l1Il.ot~lny; p_y(llioly.)ls Qo$ eseulm, ,00ld }'~]f{)W ~gLU.!llLilt ml ::;!w:t:j:I i:I.luJl:nl. agar,

. ,

".,1\,. -.,;&_r._ -~ ~- .. -

.. ' ""-

~ ~ .

,P;_ "_" ~'

- ~~,l' -

7'''R3 .A" a., Micl"OIIGap'IG ~pptlillrane. of ,fi~ ' •• «;>:;'12au)~, YlOtJln~ Cl;lU~ Ir-orn comlte.s· !~ffi [!l;EtIl 14 heurs {i~[) :trl.pp:I:IHt l'!lol1i~!Ed ,(A) and II1I~ 'f;J.~e:rra~· p€fi[r"Odlike. tB~ celJiUll~l[I" m:mplu.o:!lllgy ch,~!'l!.~estcrl a" more cooG(lid Ifol':'.m .... ,1Ul:iln ,2,g,l(l 48 ~(I'l;IllI~. MrW!JJre fOrill:;: .m.ay Cl:~~n. b6 1li1!al'D"~.!UOOs. Elr!J_U[.(b~ ~mlllillUl1lii1::!l m.:Ii,y rr,lllJ.gw@Tht iJi!lID [mas 8!liU:l eeeel d'll.Jimg: t:l:Je gr·Q\ii'!tIJ]. qrrC~e, C:" IlIikMJlfledKinltvu:n, .iCiIld.f_I"ln. ir~M~U:li ~~a :m~ be .p<Ili"ti.aH.lly ,<i:didl. f~~t 'whm .sl"joo1ed ~,Vltll a ,m.odiU!>d Ki~lY>Ulill 1jt~hl us.Eng. 2% WlI:;!SO';I, as the (looaitlti~ing ag~nt.

7-2:5, 'v.'Y J"Dl!IlI1i9c03Q1i!Ie. g;f' Gv~g1lfil1 on ·5"" .bliMP b1Oil!d ~Dli' (x1D~. IJersJulIfia Pf~01l1~ b~i!lJ:ged 00 the 1111Uii<l:med OOl1'o(;i[afm bacteria of Ce!1lLel'5 fr;J1' .Di~~se Cb'!lIIJ:.rf~. rCDC] i~NIU:p A. ~.fi'hlry' are ~J1,'ir:u::l:eri~ed by eeloaies wi:tb a m~llItou ~peut~J]:ce that is ob~r'll',£a \'\lIJlj'l iii ~j\gtl C nlir.:l'otl-cof,l'e :~qn&r Ivw 1J~\Wi:,

Y!!-*' - II .

!I~ ="

,_ -

7~~f8; Q~m I!!!.~~ ~!:!I~~Ia' iIIp\p. ~:l<1:2i1!!!!i.o.). J\mru· ... Ifr(,l ~.IX"'CI es <'.IIro branehlng, beaded, lI'il2Ii:D'len.mm:m gm;m.·pjJIsiill~'e bacilli. .Hpp~'OXiLIi1E1I.6~1~' 1 ~tljJ ii.lllll.~~um~t.t'tt:.1'hey ICillL11. alsn ~(lVe<lr o;ISCriu:;(lo!d oli'(iof(lob.aei[1l;1[ry rotoo. CEintShCiulilJ'!bet;aken ".\~le;_~ c-xml1liriilrlg 'IITI)~ Slid:es because IlflhEl £aiTIII8tail[:ltrl g I.)fOI)Crtil!:"S !)if llfi.c.lit' rnicroor:l!!ii:lll~LnS; and Oil'l]er .. cllib'i)m~·cetes"

7' ... 2B CvlQfil~ gf NiJ~.r'dta! ,iE!!I!IiI(fu.rvld.i5!~ Gilllll 111% tdii~p lJlwd ,!Iii!!U!iII!!'" .. Colljui~ illil.i<:tlJlr' appear as !;IOll~e[y.. and i?!;j: 'lhey mil:~li re lilllf'y eaa d(ivr.J1 (JIp iii number oUml()rs., mn!!:in~ rm.ll1 .:iI {:]m'Ill;y-\>\.Ih.lli~ !.O .It\!:UQ1W~l}r!illn g,G. ll:ma; red. ptxr!]!~" brown, ~1Ulrl blnt'ik. Mos:tt'.O.fol1i2::> b.[l~~u e~[iU'ly 000[:.

7-2~ Mo€l'lftBdl Km.,g~1ii ge~d~a~t ,.tI!L!r!I.~o.oonjir:r !J[JE:de.s: <'lIJlP~ar ar.:b:l·f;r~s,ti'L1hCfl ~nr,llined with B. mli1dili.ed Kin·, YOUD n Sil";miTiIl S'~T1 g 2% ~]1004 as ~h.'t dtc:olo[~zu[lg ageill. 'li'hLils !ea·· lure Ihelp~ re dis[ i I:l:guis:h this IT.tinOOlQialJlismi iErOJ11I. i:lItheF3Ctilll· O!1il}'Cle leg .

~29 (ialh::t:rn~a~i 11;;11 H~~ljj' ~~~,n:!iM'e3 IOn BAy ,aSIa!:" I'Illate .. Yel]ow-<()!'OlI!1gc oelontes ~.r J\loc.li1.rdjl:~ ll!l!)['miaes. ,]"hc' yellnw coJQI.I" I:;; l:Hore rO.l1lI!b11:0[ll wirh coio~l'es of ITIlrJIS[ Nt)- 1"IiI po rit~ s:pp.

7-taI0' ICV[vnh"'i!5! gif ll'iVell'l'Idfa .asfun:IJCIOg '!li!I'OW. ina Q" IDJ!N' Iri!'alIi~1 flo: 'lI1i!5:~. Colo:tlj.es arelarye-, dry., b:u_mp:.'f. and h''a~l;:d d·tD: 9' d<ily:s i;ut:uOO!l~ol1. at l7'lC, 'l'hL~ mQrphdl,iJID' has bee:n referred to, lIlS ~:.<1.broll:S' or,t:IJn~e riJlrn~,


7-3'11 11d;'_I~ilIIUC!in of the gemus N~mJ'B!!!!!oI1U!i l:!iiQchEHn!ea:11 ~iO!!cti~.. j\'.&!t(lnJ'irr w;p., are ite.oohk' aci:in~ Ollllyt:t! tes [h~!. r·IE:~.LlOt;: lLlirate W Ilild~e ~UjlJtl Ol'~ 1(1), hsdlVlyz:e Y!!'e:1! ~ ~ijl:w' hJ ~(!l:jil3r), and VO!IV 01] [be surface of a broth Hl~nm. such flslhe~h'l~lYcnl::Jt:e brnth (llI] U"B ;lfigiu.~.,

~33 Am'!ngi ,~!if:II 1!g~I"(!l~~1:$ rs~QtlOII'iIS, '0,'11' No,· aaf'dl'aajjlt~kfiS!Ii~, .AU <W1ill~) acid reactious are IIl~gaJl:ij'I'~ t]l1ll1~ W.[mWk:idJ~tm] iP~ah;~~ casein, nywsllJ.t:. :w.IlLWnc,~!nd by· W;.laDtbilLle.,

'7~3!2 A_m~n... ,!S!i!C~!i:l1 H:JChr~l~jjlii '!i\e~~tOnro Oif Notu:lH7lJ1 tI.t'!D'.BIBI.o.nsl9. rclm~rmC'<1Moll to tile sll~ies l!el,!ells \l:!lI,l,lll,' marl~all·theh~lsJSUr hydml!1':si:!! ot!f !lUlinom:'J&: easels, ti["VS~.e. ,l(aa:it.dm:. ;Jll.Jd!I:I).'[)CfJ'::MlItmm, CIC~V:DDg 01' the ~g<lr around tIll.! of.;t1]O[I}' ls illilel'llt'ded as ... positiver-ellctiDIITl. Both easeln a rldl tyr{t~:D1e are po:;iitj",e ll£jl.llm! rJ!1ll~ ~llrodmilt:l:} Oil tilts [pll'lc. X!.'l'l1 t.'11'l'~!l e ::I11_d .~J.1[loM'II1~hjTle (lop' jlmI jJ,gti',cmu ,rj'Mt[nUlls) ~;lre fieguHW,

7 .. S<!!I Aft!ln~ acldl 1h:r,dr'CI!~'sllSl, li"eililctlDfii91 U', 5t~m,~~.pp., }\llre.!ctions are potilli'I/'JB: VI] tb.is idenJbifit3tiGJnpla:oo. UnlIke ~MyNt\i6(,lrilll1 !!pp.., SI~p,[ll'myt'<?~sP;[J\ hydl'O~e: casein, ty:r.05jne, li:lmtl!:ri.t:ioo:, HDil LW;ptEmthIDe... CQln· UDal. 1I!1l'[p,hdl.Q&Y rn~.liIlbli!."ll the C'-hiaiL1", dry 'wht!f c(llorn:~t:;ll (l!r NO[~i!",rJj,!,! spp.. iI.JLfutlOll~Jg. S'trt(~~~~m~Qi.!'g Sl~. are 1.:!I:''ea!.pooj,tiive, [!~ are F1iUf3~@-i!'IiegO!lld1j.'e, dis!:irngtli!l!:!iilJii!!. t:he ~~ml:'<J. lW1al N;o· L'tlrrittl!>.I'.t1',

'71-35 'Diir,am I.tglli'll' ,ot·St.FeP~~i!!!!! l!!IIf1;p. I~ :'::'12I5:O~ •. .s.1n;p.[(jjij~J('CS spp. are Cll<!1racterimd tl'l.i!Cro~opi= CBUy !:iy bnlll1;t':[):LO,~. beaded, gr.m];-p(!ls.il.'i~ Iilamantous bacilli, hn!istin~lJ:ishHl::ile rr'llll1l. N(licm'dj~ spp. U I1lik~ M:'1.'ffft~m ilipp.,lhe ,ll'<lrtiiJil <J.e: id "fm;;t staln 1:'l1J ~t['\fC,

7·~7 cQ~gii'f!le!Bl ,M AAlncm1pa_ :l!IIpp .. t:x10) .• Tillli>e Ill]'e uhe eh~rELc:tl'!t-1.<jil~I: "m.oI:11" t,m'Jth." ,ooirmje!l:.:.1' AC(j'!'1~ rrI,\ji!:fll i;sr-atJ,ni ·~:hmi. d!~verrnpl in 1 to lwecb Ob1 ~~nr 111l:l1c:5 inCIl= bsted umfiCl!' :;[[lJac:rob!c C'QniUUQM:. These cilllotl!i(;J;!i ,EtEt:: 1:1II!;8iped ,,_rlld r.ailisoo ',,1th irr~Ular mnr~ii.h!!.

i"",3B 'Oiam.Clli 1fIi'!j11i!iil;i:~mliiig HiIYar 8wln af ,AlGunDm~ilII" gllJ!", '~:i':lIaIl5O:~" 1i'h~ Sillfl(:i'!\o~aiil prllplird ([mIl <I t~SS'\!i(j Ib!rl[¥.JY !ijlOOif'lfum. WII iJ]0 (lfJJ;al~r k;: u dUIJrHt:lr::risLk,: ":.'ndrm g)1f,sl1'1:.1I~;'''\\.fIiic:h ,oon!l:lsts oJ ,~ :!{l"<I1'r!!l.:Jar mlrr,oooj(l]1j!' surrounded by IPMll~£Dl[elif]jihlite. Tile A[f~mJmyci!'s spp" an: !!FdmiljJo:>.t~i'!IU· bacillLi 1:h8!t Vill'Y till ~l!!le' fromstJ.o.t:'!:, diph.tl'tcro.&d fi:lnEis. to long, fur<lJlJ:d1ilU!!:l fil!aJlle.tllllt UcilikJ.'t N(I'u~n!rn iSPP. tfu.ey am not '[)1iIT1tia~ly acid-rilS~.

'~D Iglcu":!:IlIlillm'lcBI Id:en1ilfieall!;lilll 01' .,;iIetli'lg. myos'. ,f.~UI. ·r.ltmn .fktill~myre;!i: S"fi.f.'I'. roouct mm:are [CHILi· l:riil'C ~ rtd co1(\lroo' bl"Ot~ t:i1f f'.tgttt) 1Ttcl.u.dang,1rtf:iitlHi~jDIl',S i;i;nrilij, Ai[j)rlll!1(1Thmt~: ttcWwn,!}Jc~e&' jjmdi~ .hydrCll~\'2.eS:Bscl!II:~n [bXI1(k slam 011 kEn. :~mtl1ot lattil. hliJU(I'lI!;l' simi[ ill center).

CllAPTER 8' Enterobu,cteriace.u'e

n he Ei~'tf.i~oJ)llr~e'·i(,t(,;tae' are the, most com~Dn ,bac~erial jrHJ']~lt~s en~OUtJlf>ercdlli~~ the d.l,rligl]jost~c ]rnJbolaLo:ry. They can be ~so~a~e{l from a w,lde variety of SPffC]mens either as normal U.ora (pr-octomjnm.tLy in the :int~stinru ~[a~t) or pathogens"

T~ e gram-negetive badUalli'Y microscoptc mOll"phdogy Is simi] arm 11'11 on g al~, species,

Colenles a""" I i,c;"'<u-l~'Ii ~.~,"'~ .. ,.,;,,],:1 o,,~,., '-'''1 ,:;:0.[ .:!Jl""""M n"l"Ic,,:1 ~'O"'-r:- 'D "1l1"li"1,:1 ';;"·Inn~"'" 'r •• "~

'l.J'4\I J\.J.II.iJJ, w~ _ 11:' .... ~~!\JJ" ill~, u'GLW.,e.'i;,o' {.,u. Il"I, ,:5!~,II;.!.LJ· 1"_'·UL ~ ffi ~. -'"!"I~,lf" H;}_,'I;,;.Iv!U, 1~_~:5,II;.!.L_'!! 1,\~..i.!f""'U~ ,~~~~!J,v~!" _,'ii"o..b

mentersappeae 'Pink on MC1IcConkey' agar and have a greenish metallic sheen on !Eosin methylen e blue (EMS). The E!lu'rob~~{'teriac~'ae are very active bioc.he[Jlka[~y and can be ,i.de11l.i:~edwi.th a wide \'ar~et.r of tests ... our cha racteristles (~r the EUler~ o,bll(~eri,OC'e~:e ,are' gro\~lt[llonM~lCiOQ!JkeY~lgiir,oxidase=negi1Hve., glucose fermentatlon, and reduction {I] nitrates to nitrites,

a .. 't a:.;teJ'~Db'lt!il:l'hJWoi!ljC!C3~ on 16%, slh;eO\p, btGoci iI!I(9Dt.':. (,hm~m:'teristit'col{l[l:j,1I[ mOlFpl'ml'ug:' on 5% sihe.qpoi!lJucI ~gHr :shnwing ]ar&Oll. dtg~1, gr~~', nrmh r.'11Ullyl'lc OOlfllll le'1l', l~eI'Dli)I" .~i!liS 1.0;: !,I'B ~a'ble aml '1i1{I;l Ch:lr;lc~e>tls!tJc or 'U'l,y one Genw.

B-2 P~fe",i!lO ifioo~ile{!Ij, 'On 5% 5h""""",p b:IDod ag,IOIII":. GroW'th<llp~an ltl :::PD'C::.a,d ;<JlS a ~ln", O.I~ illloe plah: Iil'~J,[[IllJil!li: uJ:.i~in.1I oolo.ri;.'l' gi ~ln;ak tim~, ulki:1. !.l&lw:,u:lillg in \\Imres. T,~l iii: char~C'~,~r.is~j;!,; 01' ProUI~rE spp" is. c.l1loo, is\''l'armiDl~ and! s Ll:gg.~slsl,h~l Ihot;; l1Ilunl]org.mH!!lll ls mom'!:>: by .IDednlS o[ iI;agt'Iia.

a .. A "Ii _DIU.ar". .pneumv-nlse On, MBI!;-CQnk>8¥ q,li!Ill'. Ra:pld lactose rerlThfmtilll:gcol,m~::>,td KMlSf.dhlJmt:u (0- 'tli!~ oppel'ln;r-1nil.:. I~r.ge, gllSlenirng, ami [13Utuid. Tbi:S~ir.JIll'll!i proh><lbly IC!lC<lp.mli21lcd !I!mJ tjluwe['tml up -eElt:!!, mneold. .1- 111OlI¥h Ibi appeamnce ls aSS'flclilllJte.m wil:IJ Ji'}-ob llJNri Pllr'Kfi,w!ia~, it is o.ollmiqllJe for thatspecies,

a-a :A" Eo eoIJ CIA M!!II~Callllkuy gg;ar. :Ka;ptd la("t:oo~ rerme:F!tillJ~_ stratns or E. mii a~M. <I.!l slimy pm oolo'Ji1if:s, Oil ~\!ia'nlJ ~ey agar. lai, E. la-all 0111, MIi!IGCQnkli!lJ' ,!!iIgjlr l,e~ii=ei"U;p~. 'C. • rHJIJ 'Clili'll ~ I.lheill!fl' bilDDciI .~gCU" l~eID&'iltt"~FiJJ,. Go]ol1ne.<i ,;rreJihiny. I)llaq_!JI.I:, ~."1"~~llL-;j;:tJllot1.-"cl. arid attaIn l ~(I 4 mm diac!.'!1ett"f oU~[Il[g!JJl,

8-5i Pigmenil:ed ,88FT"'. ...p. 'g-n Ma<i;Collil!iNy ~gpl'. These eolonies flppear red <to! ~boul.d l:10t be c'llnf!,l:Sed wIrth tile pJUlkCOl!orl:iul1! 1;0 ~<lGtus~ f(,'nll~tUati~[l ;sIwWD.i;n ,fl-l. Rilll1' Jitr<li.!1s or Sffmlict spp, ~IUll' plsmcn.~, ~;..'hIc~ i'· ~C-IfI on all solhl rnedfn Indlldi g the blood, iIIglirplaJL~

D~IEii IEAI.ri!:!lba;(;iI»rlll~. DIi'II M:aCCCiill_r¥ ,~p .. :. All il.fL'Ii!.: lac:rosc :f";tmcl1Ut:r,, and :iIilCit:o:se·:ne~l~i\i'e ¥Pe~1oe~ of th!;:

E:ruerolIl1cr<'rw'roe' appear as ,coJ)orI1r;< !i color Qr t~ medluml oot[):lIi~S 0;1'1 .M<l(;COIikey. The cnlonie or the la te I;u;:mse re~l]um'Lmg species mal' appea r r;B'~[1 til' i!;lintk 3ft.er ~ m '12 hours of In C lI\}aLilm.

,8:-a SBImo·.miJ11a '.pp. ,andl,atlill.1' ,iE'aifuRlbf,r""IurI,SG!8'S'S o:n Haik:ta11il!11 1I!1iIi1l .... a' agal'i. l:f an 0f1SW11<:mJ, 1111.*1 nul fe1l:mm.~, laCW5-'I:!!. ru.co:me, 1)[ salicin, the celontes wtll <I[l~ pear g~~1 rc~liJr ~lr the mecillu~ m I. ]f the" Fnir,:;riJ(l:rC~UlJ:Ilfn P:f(lducl'S HiS, an .il1J:llpflj"taJl~: chm:m;k:rl's.1:!Ic ,flr Ute ~'lt!?roJ1t! 'trr1- j1r'~~11l, the eelontes ,rill h" ilire bl·!ck eenters mod ~,or.lII Bl:)peaf' (;rnrnpl,ctci)' black \'!"ltil prot ngad tneubauen. 'JI'h~t' bli:lc,k c IOl~e:s rc eharacrensne of mo S j"1 rl!!!l.rlji. [I H~ alg1lr.

a"7iE>'I1lfie,ruh:lilloli5lln} CI!II;!\IiII1Ji ,an, H:~k'jO:&1!II re,""J!iI'r:I~ BgElF (MEt •. Mo;:,~L s]Jlcldlill5 or E'~I,I'e'I'DOOr;r~ri!l,~e, e:x'l;:etll fur tile l"i;ljJid l[l~L{Plii~ fernreoler.s inclt 'di g some f~htlrIddtl. ,~l(ll".slelra., IJ'ttftf ll"'lC!t'r. and ,"l~aE'r :species, appElUJr' oolor~es:s o iii: elth er !\1acCm k .:. ur to 'ill) !ll!efEnylene blue TIMB}ag.ars. fecm s:peclmen stl!Jti ld be in 0 :t!1I1 n ted onto a l'IfIOre ,;;:elcctt\'c rncdillin uch Hsf:m 1J~.lIr or X.VI05f'-lyliifle--de~,~ol,ue (XLD) agar t ~dp diffeeenttate emertc me tmal IP<JllhogelJls. Fe-J'mentatiot:1 of Iactose. SIll erose. ::JrndllHlltlWcmu 'IIlLt:.s lin yellow 01"' salmon colered ror(lIlll~ due t(D~Jb.e pH change OlD the hr1(lllfltl1]"~:Ull blue M1ld ac i& l11chsin .mdiGEI~OI!.·S,

,8-:8 Id'enUfl:c&trnDIl'II Df th-, ,EnihN'DJJacmnaeaaa' __ lfi;e rli\pl,d !IIpatlet: mtihDd. ~I~ts are comI OfIly used for the li"lllpld" (Jt'1i!501mpti\~ idEllllliJkaliou or ~.mlly rjJlki"OOrgallil3mgtou.p~. 'Fol"tblt~ melhod,,:fI'ller];l';]per or R pi'lPl!:r disk Is i:m~r.{lg.l.nltM womb. the substrate. To perrorLtl the 'r,~SI:, ,(lne or [lilG:re- eolenies are diec'l:cd ~rUh OlSl'Crilc !l\.l(lOOEin app. c8tnr ST[ck fl'mnthc des lbee! moobU'm and ,fIippliOO to tile '~eJl


S·1a O!Xild!ill!i!J!IiiI' IIillf,Kllit Nllit. SOIJ.J.t: ,wiCnlOl"~1:I11i.alUs PIIJs. simi: ei~~e:r cytoc:hr.orn(J: o.~idaJile 'Or iild.opheinoJ 'IJK~d~:;ill, ~~~llldJi 't:~I!"lYi':'I;! l.1:IiI; lr<Jim:ipud 'tlf d~dwili!l f:ro,m tionm eompounds t'N;UlH ~~Q c]icr'lro:n ft:t't!p1!J:rs M)];) •. ilI:1 this ltest. the sIltllil:mte [)llrr([lPltmlliyl~n,!5'tl;j,;i1imil:'m i[ml}F(]TochkJri:d~ s:er'!.l'ES as aD1 ''].li''~ it6d:.'CI~ eleetam llIi1:re~wr ror the ern.'0'm~ ORildl~fIC_ The ilyt:: 11<3 oyldll"oclJ Brad r!!lrms the co]o.l:'ed clJ(lrnpolllnd, l"llliol'h.ec:ol bLLKi. 1.'01' rlus 'te~tffil~!el" ]llil])Cf' Is sarur,fltcd wit~[ tbc suhstrate. Colonies 11;1'1' tbe !'!!!li::M!'£Pfilsmoobe t&il;ood Hl'e rubbed onto, t rn.effijl~el'~ap;g"1."' willi it t>H:ril.e 'L'.IGJ(ldEln 3ipphcator stick. An il]'Jrncdt~te C!)].(!Ir c:nAIl:g-r toO H deep blue itndicateiil·2Ii 1)Q!Hi\~e i~eS;t ~f;&Ul~ [~[Jl tl'tt i it!l' ·~JWlt}, thetflllmJlJ!Ti.'LeJ'.Ig(~e Ii re ~(k~·!.l:~li¥t: <til:J.d therer~r-.e i;he tneeulem sMuld Il)l):t c:l't>m~c: the .colur.of Hu; p<l{x\f (3P.;:Jt tl'JI f'lif ~~)-

11;0;12 .qj;-!n ~tIr'!il!I'p,h!. nym .. ~ ·,D,· 9 .bl4;t:(!!PiJ";!jiIIli"'l';l'~~~~ [UMPUI) ..... t. '.I.'i:Lf' eI~'1l:1e 'l:lo-galactQs;idasc m.JXIlate!51Gl.ClOSle .ib:rm.erxtatiOll," Rarpld bu::liose £ermenter.s, hm\le'll'f!t., poss~ss OJ permease C:llZ,yilliC' thaI 5Ilted:5 Un:' l'eddlml. ),1eidii1,J;: plak Cti0JlW M. M,u:£o.lfIkJ8y ilgar. Fwcluctim1 o.~ ll-gal:;Ji.cl:mid,a~e mdy ru~u['i.!; im cl~a~i:lli!I.cLl[Ise re'mil.el]t~lio .. n illn 10 (il;'lysl. Tm;; IJIOJllllll't,QOO ferm,eL'l.~~fii p.J;I!~~ :w:ll1he:r !!!n~Y!lte, The ON ~(~ 'tc:;t is a.fi:l!pld ~~t ~ 4 hl]ur~, rOl" ~-gal~CI!o5Idi1iillC. S·g.;ilnclns ldase breaks db"'1~ the s'llooj'ral-e ONP.C Illit~, OJ-nltroIDb~ny.~ • a ~l!ow {~m1;)jlJolJfld. and galactose. A ~Urn\' CIJ I er 11ld]C,fltiM II IJusi! iv!:: test 'dt1'~~ j IjbeJ !:[IId I.UI.!' ~J!.:III:II:' dJ.l-lI]~ ~ ne@B'lI'k'€) 'lil1clic.EIWS oll'U.le fl.onl!:lcOOlle fcrmm.~lcr (lj>Ji tl'.i'~i!.'h

.1i11 l ... liitO!IB 6-:-pgI·tE!81. Ill.do1cl.sol1~or lta:m d.egrm]atiMI pro(h;rob;, or the am1no acid tryptop.hrlll. [w:l(fl~ ll:rm.!ILIUl!J by II fD"I!cmo:rganIRIH grewn i [I :i i1f1(!ditlll~lI~ij f,~l iii; rich in 'ILI']'.pl:fJph.i:m C;.:j·!1 be detiX-i'OO lbjl' ~I~ lIbmtY' to com'b.ii"i.e~vi~h. OG\If~£Iif.i aCdehydes ro Ib:rm, at wlo[lQdooJ!t1,!,ouml. For the !i:po~.I'e:!it. 6)tN pa-:pcr is s.o,flktJwnh ei!t:na pru:'"firl.Ll.UetJ11yl~_rni[lOC~;a:li1il;i~dfhll'de orpari;H:lfi1ti1,elhyl!llllli'l1.obe'lJl7.~ldelwde (Kov-aICS 1re~[J/l', CO:~(lnees of the Jni~WJ.Jilirn to be' t:eSo~d are mbbed o.tIil~llI€ :~ilte~ [~.pe!:' with, OJ, ~tl;:t:il~ ·~vtl!lJdft,[l, iip'~U.o::iik:;r ::; L~!l."'~. ]fpilrndi· methyi.arnw,ociJwannald(]'lUyi:le ~ used, a blue-green ;jJ~iD.F a,p· I~ean; i:iIlmlk:ilial<;:ty in ~h:e IU)I:i5'llln~ of .ind~J]e. ~f !K,owa,c,'l flmgern!. i"li lI.st"d, a b;ril!!ll-ill pink-red color dewer(lp!;, hlt..h~ rigl!fie-"~M[lg II'il~L<lr pIi;[Jillr soaked with p~radtllle:t hy~,;:j m.l~ochl ~~Il!l.!ldehyde. Ilfu.'E! tiCllmlY on tbe ri~hl is po.~Ul'll"ewhUe the (lI"]:OO!l! 'llr.it) Jeri' ts ,rn.e~1Ili~. The pi!L1k color lsthe restl~~,of a lacIDi)C-pillSllh"·~ eolor :'ieh':i;reri rnl!ll MaC((lnllroy flg.tr.

D·'1~;;;I 1M1~"~ r~w~:e!l;.p~uluer (MRi!IP~ '!;1M, Both tests ~. poi!t[m:m.ed Irem t~ sameiJa,Qt"Ulll:ln SillS· penston, which ~:S G:livld.cd rOtt ta~~nng,. 'The mlJiBth.~'ll~ ~est IS lJ15ied. to det.enil:tt.ine d]j~pH (If il:.fue end 'I'rOOJiil.C~ of glucese rctl!iIlen:~alli:o:D1I,. ]n c.on\Ter-ting giu..oo~el(!l pyr!il~rtc ac.l:d. sumc .iH'itiU"- 0'b.7cteriQ'Ce~e ~uru add end prmlll.tl.s ~ pH ~~ Io:ml 4 .. {I!~, "in tch t~tiLI (he l1'I:e-tl'lyl red iJ:'iclil(:,tlOi[ red, wh.ile aUum; p:~a;rDe aC~l{!ij;1I (pH ~~<til~r lb,m iiO}, whit:]]. tums [he irlll;l!k·..;1B,to:r )'ellen",l. ]:1H.~ V~!i--fnl:sk.il!IJ.m: tElsl. ,del~eli!l ,::t:J;:el.oin. The- :re~genls~[J;apblb!:l!~l1rl KOIIHI, are mMed to'l:ihe 1)J;ri111 ~mpc'llsj[lll. t\cd()ifJ is u:.;-i(lLlo)ed to (li;I!I;~tyl, }·M.dtD:~ I[l red eeler (PH :groatcr than 6ll~, 'File Icll::5etof tubes shows 51 ~ith,\!:' 11m :1;IjJd EL I:l:~tire !i"'Bl (ne color ehaa gc). arrnJI ~i[l: I'ig till set ,ti,lltn,'I';S .t:I rneg!lllh'f: ~Il. and a [X.l;5~ [hie VP.

8,.11!1, P;henY.lah'lnlnJl3 dearnlmBsO t!~t 11:.&.,1:, This teszt is: based mll tfulc .pr~fl)ciJ&lk:' Ihm s'lJmetni;C[.gQr,~~llibm~ eaml, r.em!!).!.¥: an amine' ,~r(!l~~p' from tbe ,~i1J1jIlO ,8.~Ad pi1eU~i[lalaui[l.e" ml1t~~1lg In dii~ pmih.1cli«tt:J oil phe;!'i~~:lYf4IJvi:ca;cid, A microcr~iUIii:llm, gll'4il~iil:Jg ~ll a IPb!:.n:Y~<l!Limi!leagm5'l'lilu i:s t:esoocl @y' Iil,ddiL'lI;g I[ re~\· drops o:F a ~W% fClldc chh)ddt: ::iUh.lli:all. Ii. green 00101' 'right (I~I,~H lLlIJdlc<ltcs: ill posithrc t~s~: ras LLI~" 'fbI;! ShUll. reo, m.ainf: c()~otltSS w.itbJ il.lreg.1d!~hre lr:::sl ,[eSl.:alil. ~ left· MJf.~. 'T'h I:;;, 1~e.'J I~, ~lel~[lJll ln dm~[E:Illi3H[~~ ~~e tribe; Pro!1i(li;' n;,"'i,I)-,pn:;;~!:~'!"e) hnl tille other t:li~f'r:al.m~'ju"ifi'j:efllj [PAD~lleg<l'il~"e).

:g,..''I!o!I CJt~ ... util~Wo:iii, WfJ;t. S'Il:rne o[ Illl~ Eilb:roou.c· r~:.rMf hm<e tile abililty to utik r.::lbr.<!te ali a :!>Ora slJw:t:e I;l!f ti!n"bOll, I[~ I!hrus test iI oo!ll)lll! ls !ilo.,C!(lulaJted 'I:'JJ]'tCl the !illtrfm:e ()~ a dtralle sianr and the :I111eaillm;b; lneubated n'l'emighr~. al.1:; ~0" the mca i Linn I[lOJltitii!l:5 sodiiLllIIL. dlmtl.;:, 1Ij]l1110n'i1.l m scdl8" ;al"]{t b:tu'j]~lhJ'llI:lo~ blue nlIdica!:o:r, Th~ [lJ'ese nee (If ;! blue i«(I~)r 1:l1.dl1iC,ull":5 '~b'll presence 'of aIIrnLhlA;l' end P.l."f.(J)uct"<lfl:~ 01 ~tl!,'e ellnd!!: ~iIDi l UeJr tl~bi'?), T,f trne test ts negiiUi\'oI; [hm:~ ts 1l:1il colLlr ("hcu1i~e (rrnilt t!!~.It'J.,

,8-0'17 IPosltlve, pAml' and ut.'Sa, 'lUsts" four iSpeL::i~-5 d.[ '!he ,fi';t.I'!.e-ro~!t'ihlrla,(ooe are rt\J) <II_,ID,d u.rea""[;;II);is!live; rro~:eu& 'J'li,~~~li~, ~~1lj'f'U3 VI Llllwis, .~J'{jfU.~lril .lJ!U'.f£h~'~l.i. imdl'h:r~dBJlfif! Ilfllilf'n, Ifluc isol;Prl~ L{;S~l:~ prmtl'il,lc(!(_-d these rC'i'1(!1illl'li) alilt'! ",I~;l(, :S1ll/,21l1"rltcd.[ln blomB :lI;g."':I r:', ,~I ''I.'II'01.llclh.e ~ PrtltLIJS spp, ,\ sl1(1t Indole 're~l: 'wm;~lrnidB:f.1ti fydl~c S[lU:~e5:: .P1'~t:Ui\: ~rJrm'billi~~ ]}l, lndole-negaDhiljEU~d PrOMI:!: 'I'1tWllf is Is irtu:lo~~-pos'UlI\lCl'. TiJ1.€ ,c1lriL~,.c' t~5L would rnItTeD'c!l tiOite t!h~' nD~swm:[Uers~ ~'i,1.{lrO!l'f,!,dJIl 1~'J(.!}'I1«.ffIJ ~ ,('itcal~l'De,g,l;lLli\'"e m:Jdi?rov~dfm;i~~ ~'dlJ]c.f:J ,DS eltrai:eoPIlJ5ilti~e.

&,,18 UPD!31W1! t~,t. l.'lkmcrgmlisms ttlliftt. p05sess the en:z.ymc urease h,~Id!i:1ollt';illt: urea to <J.infl'iollia ,~ild CO~. The pli!o:n61 red ialdiCator tumscense {dkllrk piJ"l,k1 ill tile presence oflhe alk'il.~ine ei'id iti"'OO:~en:s. This, Iigl.1t,t! Sht!'!.~1S ~m''1Ci.fiOCil,l t:;aedl fhri:s· l:eDlsern'sllllrea shmts that 'Were 1 Flc:uh0in-.!3[J m'f'rl'llght. J1he I~ft ii:lll'lld !'l'Iod,d II:! i u bcs 1:iH-~ pt}.'i'lllivc. The lell I.ulbrn- 'W.f.H!! inoeula tad ~",fJUl I-'r,Qw,rl~ nrjraMtJs. ,<J, rCl,old!J.liCa ~\vdr.(j;l;r~cr (2 to ·<Jllli(! lit'S). the mrLi~l:!le tube W.L'I!5. i1l1ot:llIl,ated\'\~it.h K (~!.tsi~,lJfil p}l~!mlOPli'Pl:~>. 0Ii. ~h:)W urea I~JliO'hl~tt t 1 ,8 ~u.24 hio!:.l..Tsl, and the ti!;l~ It]b~ W~ wDColated ~~lLtb Il, cill!.8i '1!l1"e1l~·[l!eQ:Rt!,'C .1n:kl~O'l)rgan ~I]).

""or ! 1"1 lill.\l l Qfla~~iii1 -1"'-10



S"'1:B m'eea1rtio'X)iIIli. ·dltlydrGillUiII8 'i'&:"ts., 11le det'~y]~clll:.Ym~n:muws CFIlfb!J(Kyl. ~rQJU[p,s frOI!ll the a[llil1i~ 8I.C~ds ~,%il1~ !lind ClIlll.tl"l:i1ina. l11e dt~h:!ir(l:m];aJre em:yrne j'emOl'eS: il\l (: ::1I:Ib(iN~'l gJl'l!lu.pi'r~l]1 :li.Vg~~LL1.e. Glucose b rt~' witbol.l;t the ounino l!lCid (.rd' [tdJe; ~~,!UroO Oincll mbe$ oot1lrunl'll:Qi glrLj~()!1e [l'1\~~ the:~rJli:rlO a{:~!:l sebsrratcs are bl1t1ellJlat(;-d. 1J!(;:tarbw}'kL" non and d:1hydIolat1o.tl are anaereblc ir,(raelions, so Ihe .LI:lOCIli~ I<l.h .. 't~ il'LDOf::S UJi!llst 1Jt' [ll!.'l.)rl .. id witch sb!t!dle 1]~i[BemllJill[) ~Clri:dl1l:le ~ir. lD[i,~~a.lJJ', 811 broths turn }1'C110\!I' due t,[I ,a:Cil{lliliC'~lt(m or the indicator tfu:ml"ucre!lL'll purple] by the' OiIckl enil ~l cts or gluCQSC 1;~rm_en~O!lllQ!l!.l[, the amine add ls doc<lrrboxyl,woo,. the alkaltue cild I_)l:'i}{lu.ct causes the Wt1icaw.i hlill'c\i'crl ttl;fuJi ~IIHdiJle pH tpllfpJe). 1111 til iS~gLiir,!J. flrglr.l~ no {Slicm.ul (iBmJ~ IMfLlllI:I'ld {iI[['IJi~Jlljtu'l ,(ii'iglJ.a mile) are ]rO.!i1t11;·~'U1f:UJ ~Yl:lun:~ is ·negative.

B~'1iB' Triple S~5181" III'i!l:m rm~l~ agal"~ranil: ;;;Inti rlb1 !tube. 1'S1 <:I,gar detects f:ermellla!~~()I1!1 or Will cese, lactase ood S:lU.CTOO~, U;l:S f?1'w.udi~l1. and gas rorm"ti(l!j1, Ih~ ,lli!.:di.mll Wll!t':<tlns one ];)<lirl gt'u'CO\\lC W 10 psrrs IAlI!Ltlilsl:: i!ilIld 511ili~~:Se. ret-· doc; dLL11it"luniulll dlr,iillhr" aud plum.ol ir,etII indk:<'u[[n. If lin: mi['T!:.lOr;giil!I1I:S1ll ff~TlJil1litlli !illlmOfi~ alone, the lI;i:rU.ire tl!)l,l)f;' t~s:laJ:n and ilutt) win tum actd l:rdll~w': :i:I{]IWe'li{1f. tl~f. I:l11icIil'l1<[g.'l!TIl:m1 ~vmble"BU!] to ,r!~gfac..le p.'fnie~m: on :H~_!;J .::lli!i:;) r su rlaee. re8IJltll:r~g in a ~hHn;e ,el!l.(llPmfll!.!)lB a 1];(1 H pillk I'lt.aJrtl. If 1~!Ctr.DS{' ill:JJc.!/i!J,r· suerose i@1"C fermented, 'l~u= entire ltlbe r.crllaLrn:sj''CUow !J~l:ImieoOr Lila'!l' grotlK'1' ClafllG[lntr.EU~()1'I ordites~ ;cuJ[bo:irny,drl:loos. [-[.!Sreac rs willi :Iiei':iric :uI'i'I'II~{!~ium ciLr<1tiC: 00 pxodute Q Ibj~(kc.i1d ptOO!i.OCl, fCf'ffO'l..l5 sl1lfldc. Gas fOli"mit"ilri,O!c:lI is dC'~W,m:i !by 'llKi 1JEe~IJi(:e ,of biflbb]~5 or cracks in tb~ agar; '[he :rlb'lwEle (Re:b'uf!'l.l~hll't"atnLies, Lenexa. Ka:Dl. ~ .~.!l a m~!l.l):I 8.1 cnmmercia l identifi'C>li.tiotil :S}'5- i.:e~1.[~1ve reaeuons can occur ,in ~he lI.I;pp~:r' .paZ-irian or H1Ll iiIr:sl tLJU!.::; g~ !JnjSC and lactose r.errLl,~[lt';:i:IiQ~. HIS pmdLle[:roll, gas [UflIliil.llh.m.;, and lihlmyJ<liI;a[l lue clealU1~illlll:tkm. Lysine derarbOO;:~IIi:itio.lll OCUIiJ!i [n ~.h!e Io\!;~r 10111 a! erob'lc " portlon. fu thts exa!U[l)e, gtuco~e is ff':mumledan:d iH,2S l~ ,I)!.'oduc;ed iIl .: ~ 118 TS~ (left sIal'l). t~~ yellllw color tndlcaung gillC05C f!lnu:cl:':lLaJl::i.on is masked by IUlc bktckreod prodlJ:C1 o~ Hz..~ pJ'(JdU.cliOJ![, "I't'i.c·smiti~ reaeuoa ,fii:lV~!lfS ilIlltm [ipPill'" p:;.rl.im'ilI..lLlllJtl rlib! tube: ~l'sim: ls pos]!l1j,le i[l~f~ti !·Cilwer pDr1iG1rr~ .• This r"f!utl.iun l!l u:g;_~st5 SlIhlulrrlilt!JIl! SPJ),

a.zgl __ IllaHII ~(;tiO.lfi' •• 1.11~ Ul TSlllioo!i h<.9J1I'e the

'f{JIlI{f~rtlltg ttlEtocm.s, (I'ef! I:tJ righil):

iSjLiIt htI: N,~S (i.",rgrmi!!'!Bn
#1 :mold Reid 11
#2 a dl acid +
#3 illkaU11B ,acid o +
#~ alkaline .add (J 0
'II" alkalim acid o I)
#6 alkaline add 0 u
#i alkaline at;[>d 0 +
;i\jg .add Itti~ll + I)
#9 allcIIlUne add Sli1l:ht 0
f4:UII ,a.lkallin~ a~t! + +
'"fI.o:::id - _".",QI,lI'!'; dk.rliJae. '" ~;lI:IlS.l1'J'l1ll1\'B =b:l. ::;~ era,,!;$. bu.'il{lle;. B-21 l"iD'ii reac"llvmI. Thl:l ?ighl r.lbllUhes hm'e the ((ll· lowmg r,",,· [1011 s":

:l5lalnit B!itft ";oS, Q~~ 'tomaiil:ln Pd' ~.nl!l'
'j!j!] iloid acid, 0 0 10
tn a lkilliur: ;Jed 0 [I 0 0
it,] alkilliJl~ acid, I) + 0 to
'#4 alkidill!:! aeld 11 0 0 "'"
#~ iiIIlkaiinlil adrl 0 0 + 0
:;;& ,ttJ[II;;,tlliullll acid + n i- II
.fin all!(:;MElti! acid S:~~gbt D 0
'JiB all!(:;ELlliiIe acid + + 0
",'.1:111 "" rG1kIw;: A]~nllnli "'l"1ld: 11~~ti.vc -1:!L:u:.I:=&JUI ~ ~lMfh.I!lljl:il:lh!>.:
IP'hI;) UJiIY.:iIIh'C = ibfmo.m; ly~n!'!fIo!l'9tlW! i"l'd. · ,

8·2'31 CliTIRH6M med urn and ~~cl:iQnlll!!', The IIlIpper !aerlJhir) [li:rrtinn oX Ibis mcl.lhn!l'l ia fu:l."d~mom;;tMIHon or (lll'r.ml C ~mhYAtjan • and the I",we (i!lllaerubicJp rlion', - tn detet.1. rhamn .:;.e ft"IIltil:llllbuniof't

The row' tT,mrI'[A,'V,!' L1Ib~".l1 hi:!l.1'4.'! thill' lfO:~~D'~Vlilrlg r~:lI{:ltDr:! ~"':

ICtlrate lI_hamIU!'-.
i;il I)
#2 0 +
:it + +
#4 + 0 'LlmILC~l?C'!'llh,j!' • Nul!': clrrme.l)('I!iwtnl' ': JlICl'fi: rhlilllTl~]IU~lI'I/C .. }rlI ·;rlliIlIIlllO~-llfllUU;,"· I!.l~j"

".22 r,tb,- mecilium iIIl'ld I!'1IBGticnl'l1 '" Thl Ll!,lbe has iIll!I u,ppcr (<i~-olAoc: I :a!llcl ,~O'we:r. (al~<J;erobjot:J ll(;Jrl~[)Il. Iudale ts inler()rcilOO in tine l~pper eornen and lJ[u.iHiI:llle dB~"all"b[li\)rIRl"('): ami !llOiW:t:· arc inteepreted ~ lhe':tQ'Wl!f podiu!l,,'1,imj,s. Lube Is sunl~ar 10 the ,con\!ellill(n:)j~1 ~,[~O [mDli l~ty·iTlddLe-l:u(llth~nl.:' moolwn,

T:he (Our rJb1lLilie.s hown ha 'C tile rollo\,,im,g rear;tiow":

Ill'!cd'!l~!i! IQmh:htr.!e
In + "I'
#2 "IF
fn + 0 +-
#4 (I + o Ina ".I"~ili1,,,,, .. m1 ~+ !.iillhu !'Llrilll'l! ur lhi; ~llJlf IIi~n, ~.!rI'ULh [I pool.liw:-pl:tr;r.'" mi. lnr.Jtlli!!I ~Lt.'vt: = LtJrbld mli'l.um widl" 1l;.J1.l1 ~Bib 1IIIll




D-G!S: ,Ci1Iali\:lIIctaftilUa lNI'iiJJofl'QJlIIIIO of ,S\II'Ifron.lla ~l.

alk/acld sii!_!!ht ll:;!~ T

e.lktacLd slng,htH.!S + .l)'~ill~+ l;~WO

ornithine I} Ofo


i:t1.d,Qle 0

&,,24, Cha:raiClel"ill!l;b l"eaeiJif.on1l!,c:d' SalmoJ!I'f!'JIB 'lIflP~ ~:'!iiI!!Jiu;;!I!ii!pil::SalmC!nlli!iilla ~P'h'lll

,l'Illltl,u;iJ(i, H,!!H

<.iJ!l:i;mcLcll H2S +, IysllllC T mDO

om~lhh1emotile 'iHook: 0


,INn" CJiMnltc'i!lil~ NilCitklnIll uI'Yenlilf.n'a liI.n"' '~IL

,ttlikJ'actd H2S,O

<11It1 ilrul 1}1:!:iru:: 1]1 H2SO PAn 0

armt~t lDioll motll~ Imdlo:le: +


8.'28 'C!If!l!Im!l;b,!I1at1'G: I'8!E!!ctlo:n. ,01 I!!lVIeull! '1(<<,3' Sari' .. ,


<Ir:Mla:dd rIaS+

n lkt'1c1d itl.S "'rAID'+

Ivgl~e 0

ar:nlf],11fI e 10 :Plo'l:lle fnClole +

8',28 'Clh!::lilt"::I;ctll!i",I:~E~ 1i'~.;U;;!U(.if!f.i W ,S.fd.e11a I~P.

,~.Jac:irl 1-]2S {I

alkiacic:l ]j'~lrll.e I] H:!SO ['>,;'\.DO

o.mithiue 0 tlO'lllllolile lndule (]


8"'28 Clilat'<!ileHttil.tr,e IraDctkl!m,., at' E~ ,GClIJ 11iI.limgl MICH •• DJ IliUI'p" 'l"~eM[CR0211!) (OJ:~~I !'Ion ~knlka,[)11rh~l!Il, N.C., .. ltrlip mntilill~ J 5; bi,oct~,eDllILcruJl tests rilr the r.<ii;licl, id~11:i6t;a1l:i!!lIII (4 ]JolJirs) Glrlhe Jlrl'~~[·!~,im;!!·m\. Th8 :;-y,si.-~m i~ bl!l~ om t~u~ p:ril:tclple that the heavy i[IDl,"U~!,I111l :,l1"L"Jfe:m~i,m n[U:li~ ,01'g;;!nli:sm'~ IJ@ tes~~d (l[mtaillshigl].le~Iil~~ Il'r pre:ror.TIled ,~n?,yll~Bs. 'ri~e le51~ illclmted il]~be !l~!me:m are !!;loges-F'l'Ol'lkmJer (VIP). lIl:itmtl:l ,(N), ,1~lfIelil,yfl"DL'lInlTlie de<lmlmme tP!)), H~\),~r.ldol~ III',· m'!1~thille lOin ,<JIUli:llysil1c decru;I:lOZY~'lIJHl ~LO). malonate (M1. I.llii!!;j.llI). escl!lJifJ ~m, ONPG. ~_lL',dlll:!r!llse' lAB:AH). 1uhm.iwl {AtJON)" Illo:>.1tol ~lli!lJS)" ,fl'rld wrlJlwl {~t~~~ &l.l, l'h'C tClilt M:~lJJI;; are I!Uer,t'I1'ctoo ~Sll}lllj~Oi)d~ [!11l11tT'ical!~1, .ood 0OO1P.bll'(;-d. ttl Idtmtilia!tiol] tebles or the ~iI1C!~O .. Jn [lIooftl~ code book trll make m~ Idcfl!;lftcarl'IJJjI,.·J11~ II1!cij:}:r.("',nl,(i1'l cnr [his 'el!<lmp!e Is:

!liP' tjl PI! Jj~ Q~ !".1I!11 Nil 1.11 II! owg,;, MM, gflli IM.CUi!, :NRD,
0 + 0 0 .... + -t 0 0 0 + + 0 IJ (I lite jdenti'nC<Jiti.an, is: ,f,'.d aud the' p:~le ,CQd~ tlI:(.Im~ber' is 1:3430. The c(I1de was: derl~'ed '[)]I' divldltlg lie tests wto 5 gool!l:!l'S ~r tru-eeand:a:s:s;lgn1l'i~~.a:sC'~e t~~,n.l'lw 6:rsUes~o:r ea>::.h setts ghret\ a~ec[ 4. LI1(: ~(m,t] rest is:;usosiCrteda;1. Ilnd the'tjl)ird test is OiIiilSigned. a L If .m.yQU thetests @N :po-sil~v~. it li)g'!rvc1'l us IiC~.rc, iJul it reeelves a Oir ill!] negative, The Drlollnbe:r::; in a set call .c'lll~ rmm U to 7.,.

& .. 310 C hi8t"a~lI'mstr'Cl f\8i111QtiGl1. 0JIi1 ,E;,~iOn UI.ililg 018 .AlP. :2D! e stri,p., '.I'he APl~(]E s:~:ril~ (bioMeri.ellK 't!.l:~, 1']H2leh"lOOd, MI;D. ~ i~ 8!. s-elr",cr;mlaiEleil ~'SlelI!, !:If J;O mic!.'i;lt:u~ of ae-bydraledJ SllIt1'lt:tra.tes. a rni[J/i8~:u.l.'ii!led 'J01"stan of COll"'ellti.Q(Jl"~ p11'ofe!Jun~, d.esigI.teiil f'!Xr O1I'em~gh:l. illCUlb::r1 lou. kfei11tifikaUoll is made ~l ii1dding fiJec~~J' re"'~I1~:S OI!ld then vicSUOliIl~f ~n~f'eti~g the results. Thetests ~-~d~d'cd Lt"I, tbili: :s,;),,:srem. are; ONfC;. a rgl!llin!HUhy~h:CI[a5e IA[I(' l.lY.sme [UJe) i"lJld !lmiLl1L~ (ODel d'«<tt'bif..:yl<lS>e" cilr'dt;.: lelT), H~ S!, uil.f.li (il'R!';). h'7~w",fi~n d"'i'lillflli:l~e j'J'I)',,\~. in(lQle' Ur.:Ul.~\)~s-P[~~I.!t:':l' (VP'~. sdatii~ ~'lr!n~. lPm;US[l: (Gl.m. InrumilO~ ~Mfu\l), ill05iwl, [lNOI. :sorlil,to.l ISOII)" r1l;ffi.nrll'J(I~6 llUM,) , sucrose 'lSiIlC). melibio.se (~'~I1.L,I, .. rrlJ'gd!<~lu!! UtM,'i I, ;l!ml ru"Otbml)~ 11111M. ,Au tlX,kJ)-itSe' ~~l ,mIlUS~, 'k ~I,:II'J'!Jil"H1t:{J ~t:p<.U",<t:IW;.,.. :Ktlilu~rlt;iiIlI >t:m.li[JIg o[ Ile5U 111': O!!llows ~om;W!t!1eJil;;:ed i.[]tel'p.r~taiiim. or p;lItJI~ns; lists Qf ~'I' Illd, ace avallablc UI~ ,I I:m:leoool!.: orin 1.'VllWlIlll!riud (oem, '.rlle lIlilerprel:aUl)u o:f this .;xample is;

(liNN ,LDC !.De: 'ODe em 1tiI~ IlIR!! IN, IHI !lliP !lli:I!!!I. .::,-y tPJ! i~ sgWi lil!H!l ~G ,"~!i, lIo_N!:Ii ~
+ 0 + + 0 I{) 0 (I -I- (D I) T u + + 0 + 0. -+ The idcnliilctllton ts £'. ;fil~ and tlht :pFOOie ccOde mmilier is "514455.2, The code ¥'I'll!> dt:rhneJ~' di1i'idiJ:lg the' ~r,;:;I;:; iuw '7 gr-.oujM, or ilII"'!JG·I'UiiJid.asr.: reacttou if!; till[': ti1ird res'! iII. LIre lasL scn ami a:;;sii!lJl!ing a scare to €BJch, '11]~ ~ir[:sl ~~'St of each set is givena 1 , lfuc second I.e:>! i:i: it515ig,m:ld a 2 BInd the third lest is a§$i!;:ll~d<l 4. ~rr am,}, of the l~t:s ar~ .PG.l1iti\1€, i!'ii!l gi¥EL1 its: SCOO!ll.flM.lii;l(tii1res It 0 if it ifll]~~,'Ij!ic.l'h@ tat!}1 ~ores: in a set cain ~I'!gf)! lrem u te 1.


8-3''lI1 CI"iaI!"aetljtr1atJ~ reiili!e·tD'g,n3 l!d' Er:INi~DaRe>,!" ~Ji!e9 ,tap'. ,Hnd Entel!'C!lNtlGi"tel." ~~!iii!i!f;i!lI~' '(!i:lg-~m~ ~ing API ;!i!g Est~ip~" '1~~urll'imrpr"1l1'aihnHlf thest>lil:Sl.'l in ri'ils(''11t'1m[llllii~:

!QNPG, AD~ LlD'C QDe ,CIFf' IoI~Si ~RIIii TIlA INDI '!iI'11i' GEn., QUI IB:N 1l1li00 $JQ,1!' RI!I4 ~c: "'!iiI., Mf!I' .. MU!,
r.:drn + '1- (I 1- '1- n 0 0' G -I. tdn (} + + I) + + + + +
F. aer " I] + -t + 11 n n en + K::ICIi n + , ...... -;- + + -;- ~·he.[IlII1I!lWeC'l1dE' f,lm~~~p.r r!lJr It I~WI1!:t~~ U\ ilo) ts n05S7]. ThB ~)J\nfflllie ["ode mHU]j~r rinr~. ru'I'(li1f'.I]';-': W. ner] If: 5~,O'i7f~_

The m'lil)[ dlrrererace~ beHwc!1! the Iwo spedes 0!irC the reacnons w~t~ ElII',ll.lM1JTIIC, Iyshnc, andtnesnol,


I 1

8-32 ChBl"~rt 'DC! IIl'aM!itrOIlill& i1!Jf' ~~li'lWmlr; b"lB: t:to,P. ·a!l1!d P.rohiliU.B vufga ill! OJOUom~1 using APi ,i!iD IE ~a. The 1l'1terprelaon of rU ese '~esl5 in. this~ a mpk' hi;

OI'!l~CIi .AD!: IlL De: 'RC: ellt I!i,..s UIU!' 'TiD 11'I1i' 1;!p I~S <t!'~!lI1 IMU IIM:C S!!iFl IUiliA -c rlYl!L ItlWll' ,AM
P"\,~I~ [) II n 0 + + (]I Fll"ul o "'" ) l) o (l + £I .. 0
P IIIjr· o Or _,_ + + + 0 P mlr U -I- 0 IJ 0 0 0 0 ~1 The protllc oon;e number for l~ Ijlllttil'lll1:1' ~ r \It~n i.s 087-1 eH.J , The pwlile C'.O!rle number lor l~ ftli.rnliiJJB ~ P mk II ns () 'i' NOlln_

'It c ililftrtlJC'a betw~l;!j: the !,W"o ,ilp~"t"il'l1> are the reactlons wH h nmlthlrtc, ifU]uje, sucrose, and ~:mygd.tlln.

B,.;33 C.al'Q~~I"I;MrG ".80001'1. 01' Sellill':sllll mlln:;: •• etA ,(toP). &nti FoDbao'tu.r' aeJ'V99.I't'IIB l~ml'ddlIB,'I' and Kfl!illlal'SU.aJ wqr~a ~ib!:d'tom~ lIMing .AJF.FoI aD Efrl'pB.

ftDC CIT IH,.s 11Ft! 'iIi1D'A ltim ¥P 1N:!l!I sa, AHA, ~SAc MEL A'M'i'.ARA
+ + a II [} U .... S mar T + + n + (I
(I -'- + + 0 0 (J o "l- .E aer I) + + oj> + + + +
({[lIty n n + 0 U {J K!i»l.1 0 + + + + + + I -I ']i'llit!' !,(-unl (."cde num ber ior ,'ii'li'm1!lll J'i'J'{in't'Sl-ms S m.B1") is 5 3U 1-; 13, nil' prdlil.e coil'e number rC'i1" f., (1tr~~t'J.lts (E acr] is 5305773.

rhC! p:rooll.e cOO~ Il m her ror Kfro 'LeHt.T. oxy,tr:x"rI{ K oxy I is 5.H 57 :-'"3.

Th~ lIi.fferencf'"~, berween So. !IIllter wu: and e. rTtI\l!1fUt ~Lr;: [lilt: rum~n ions wUIl ~1 .. tln, rhamn U se. tLrld. mel i blose, The dtl!rrr~I1C."i'_'i: betwrefJ K, (Il:'Y~M:1 and 1:.'. al"ro,~'l'es are thr.: I"C.tII.'li.ll[lS ·l'IIith om!th~l]e and ~bl(l'{ll:e,


,S-3d, 'Clml'illlcdel'lstla 1~la!iiis Gt' R~.I'a l~~~ i!lJniiiIJ 5li"9iilitW' :fiin~."ii:la· Irlg •• :20 E '.tI1IU"

- .am 1;5,1_ Ell 1!III1~"" - .., I =

Q~~QI ,,IU)C: lI!.N He IC~lif' HzS, lURE TID" liND 'VIF !:lEI!. iQLlI!i II'IJ\H U1Q SOfl !1!'!iI~ S~g 1M!;!!" .AllWI!' AmI.
'i est w+m () (1 [J n n + 0 n [I ¥cliJL n + + n + {] + II 0 +
800n ~'!.'+m ('I [] "T' 0 n '0 0 0 o SS{lll! !I + + n o -I- (l n n + The !PIt,~il~ .orJrlBrllUmiJefo fot Yt'fl,Jr!kl ("ju(;'ro(~ii'iC!l W I:!IU) i5 1014 j i,L. ThB pru!ilEl coda nmNIJ~~dll[ Shii'jjI~rra :sormd ~ S ;So[i) ii 1 WiD:2.

Tho i'JilYlllren.cil!s hetween II:fila '['!i'IqjI spe~~ :m~ 'tH1e 'lI1$:iIc!'iDl!l:!l with Ofr.llilh:lI1G. UrM, oorblml. rbamnese, iI!I:l!c1 ~l,l)cr(l(le,




:D .. 3~ !Dhm'"it~!1!1!;g!!;; I!i'@~!~~~ M ~,~~~ mll'BbJ'tls ~ta,pi~~ ,imDoJ"'g'8nafla' ,m~"il (mI1d:dI!aI, !andl rP.n:ItV!UiI5!!i nIJprl:!l!' ~J;,tg~~ 1"'~I"'9i !!lP~ ,2Q IE _rips"

!QiliiPC ,l1Dir: IUle ODe ,el'lli' H~ IJIRlE, TDA INDI ViP tor;!. CIJ! • .!J .. AiMI !NiiI! ,8~iI: RI-l' SAC MH.rlMY' !!lb\
F,mll' 0 0 0 =!i- + + + i- II 1[1 Prnir 0 -+ o 0 I) I] 0 0 0 U
Mmor I) U U 01- 0 ~ ) -+ + + C] Mmgr 0 + flr flt 0 n n 0 0 0
'r \'lJl. (I 0 0 (I 10- -'I ·1' 0 Pvul D -IF 0 0 o I} 0 + 0 'fn.e: pro:ffte code n~!lmber rOl: .i~ m i~bjJ'j& ~[;> !Wir)'.i& 07 nom]. 'Flit: pm:l1~ ~ lIJumbt:[ [Oi' M. m~~Si ,1M IDQor) if 0174000. ']'ta~ IJwllL'c rotllf.: II'U1ruIJcr rot. Po !'u/.£ji1!'.is (P!i'I..l~ is: 011 7 ~O~ 1.

'T!'!.e cl]rr~rem(e:s b~tWll(rJl .M. rrirlJ"tfjillii <lliHllllljmbm~.are the reaenons ,,,;th C'iiffi1>e.. 1'] ~8, and indole'.

TI~ d.irrlfi[em:~~ ooL!;lI;'l:iIi:IJI.M. mOJ'gltrril <u uJ P. iilH1om"is are t:lle reacuoas wtih l(rm~tlilliHle. 'ci!J~a:te., ]::I.::;:S, ~r'("-o:lIll,mu!1 illiI;\)'id alin,




,a.,3B Cum--PElriina 00I01Tl11l1 mcwphu1'Ogy wlIlh !blOC ---m~1 Ilda:nUif1!aaHon. An imponoml suep lrill[il!!lliblaJ iih.lIlltilficaliQn (If ~11l ii5o:1at~ is to compaee the bilJchc.ml-c~1 re;mt!i.l!ln ,-nth 'the c:olt.i:llia1 merpl1-o~_Qgy am:! reaenonscn the [lrirn:uy pia1illg [Ofilia. ill tWs exUJiple;, the coloraes of: :~{!rl'mll~cl1n spp" 1m::

E-~2S"pOSmve DIll eh . -IRE a~9f' and the ":I is alsu tJ'Osith~ on the - P'JO E.

Cl-lAPTER 9 Other Gram-Neqaiive

M" ..


D he gram-negative microorganims pre nted in tIns section am those that arc n l i r:n the lamlly Ei [t rmbActerju'rl:!'llI'€. Included are the .1 _'i~'S tj'a and NJoraxf !I'a species, misceUaneous lastldious gram-negatlss bacilli (Haen'!Op~1 ilus;, Bon:lrl£j![~, B:m-

rdltT, H~·sf.gundl[~, ,f3~ktm,!J~1,'fJi Kin"qeU~ . and rli'pr~o['t!t~uphilBi7. spp.], the curved or commashaped gram-negarise bacilli {tCnmpy1o}Jacter ,ElJrrldJ '~libri,o sPP,J. and the aoefermeutatlve gram -negative bad Ui (.P'SCI,j rJ:mmma.'I'., $p'~.iuBomar.!'as. Steuo~ ropllwmol'.ll1s. Slu:w~ul\eu.a ... dM€to.bacu~l: and r!ll~ tmr.f.{"'rfum pp.) •





8-'11 Dram siBln Of ,NetBs'rlD .F.I\P~ r::mm 'mined , me r ,of i:I blood culture slllm'nn& gftlJn'_nega~i\i\e tl!ipJooocxi Ch~irddtl!r]ilic of Nf'i.r.wria SJlp. The- ad)O]'cllml ~irle.~ of the cdl .~alin look llatlened OT kidney bean·Rh:J~~d'. Tlu:,d1p].oceOlidd<t;,fu'l restst d!~ej]I.o1f'i7..lll:1itrU1 and, asa resu It mOlY staln ,~UI:\'JjHrMi(lble or grlml"f1O.'iH:we (x 11';O).

8-2 Neffi8-erla .im'en'''gltldl's alii chOGiI!llUlt ".Oli'lo .'\h·ls~MirT ftlf'11Jfl,gHifI'· rolOrlLh:s uri i:I d]lu':1lJ1Hl.e ~Lr p!Hlle lacubal'l.:d l J,~"C I~l 5,'''.'0_ and 55% to fin', llmmiEln.tiy, I'he colo Ik::.: Wl'e mnre :gray thast yeJltlW and IliIlT.lnetrnalytDc, characrert Lies, libm distingu' h the pal.hogenit:: spede5 of Nelseti'l trem mC9[ of the '1l:!lJl1p3th~et1ic spedes, tn older cllimres (" 8 hUUflSJ. :ill greenlsh ca 1 can l3?pBl:l:rbel'l~Jth the celorues In the: urea of hoo'i'J grm\/illh,

81.3 N~"IISA.t'ih!I' mAn:1"'SltfmA Gn, ,5% ISh8ii!!!ip' Itll!I!MXI Itgi!l~ Ci::rlDl1les2lDie 1:'0611110. smooth, opaque. glHs:tol]ing. and ] i:f,I 1 .. 5, ml"ll 1 ~'l diimnet'Cr~ tile celenres aee gr<lyl~b lij color and Cain lII.ppeax Ilinkish, as sJJ,~\"iI:], mtm.s: 6gur,(,""

9-5 ,M'cH'1I!I.~1JJ'la j,BI"8Ii1bama"a,' ~;I~FrliI'~Q ,101111 5% .'h,!I)4;IP lbh;.pd I\gar. MG1'{lj."ll~la c~t(lrri~~lii:s gWl~, well on routtne 1000borawry enriched tltetua. iir.H: ludB:n.g Mood and clL()1;;tda;~e .. ~ mJ\:,:.,jjiO!l. Cululli~t; are iJL:slLklillly Op,ifqu~,! rcund. and li'l'h'l~e:, Mk~WiKUpkdlly. li1e_;1l' IJISif btl '~:mfLLSBd\'tIilb .rl!O.[l" palh'ClIg_eniC' !It[::I'un~ of Nld"'JWP',!1ll Sllp" L~~lil' they ,21re as- 611or,h~roi)~'rt1c m ~<!rlli11!}~-dr<lte !ltl.lI~<E!!U!J11 t651[:5. th~y reduce NOli" and, unlike N~!sserII1 sp;P,., !1J:f;Y tpmdi,[ooDNal:ie.

9,"4 ,Ne1~!ieii'UiI pii\l~rrftoe~e 00:" n1'ad:IDlIIod TIi~""Mt!li1I~ ~Mi'iJ'M~ 8g8r. ~ITM is i:l :sdceU'imi!i!l]Hii' mcd Lt~1fI fOr the isoliiliti«11 of N, gajUirtill~tl<,f'. h: COQ[I tains allJUUJ!l]j~ oerolJ.i;al <lg€.['lrrs: I va tlOOI]':J}cjn. ,aQ~t'i~in" r.I}'S'~atill::l" I:lJm:l, trnnethcprilLU ~a~~.aM tj] ~nl1Hbj tg!J;03 m"positi,IOoe and other gro'll!~1-neg.]Itj!ili.'i !'l1jil;rOOJ:",~n1lsIlls. molds, some: :lIeast, and ~]}~ swartmng or ,Pro.!'f'US: Sp[JI. Aller ovom ~gllt i I!1c!Jlbmwn .u~ ],:5 "C: ,ijJ'j, 5% O);l' COhYlfliflS iBm :!iIl'La~l (IJ,.5 to 1.Ommi:l:l dimllcrer). gr~y; glislocllIng!. HIlI:li OpaqjUE, 'jjU1Y if])CHl3~ in ~I:ze lJIlir.1h prnl.r.m ~ II1C1JIi'.!~ D.iillTL CJ,jlIl1Jre~ should be :b.el'd for f1 bAJiW"S before reportlng as '!licg~U'l;.'e.

u-i6: II~nt!il1f,(j:tI!"f:gl~ ~~ N~j~~) !!i!!PP." b(J '!'.llru'ba'" hydr8"~g li!ill~galbll!!liI!. TI:D.C standard ,uJ:t::'Hu.m.l Iar trJe[lilll'iu!;l. .n.,~~;''jSer;~ spp, Js t[) dJe:tcnllli.m:l acld In C1'J\ {c:ySlci rrJ;e..II'Y[l!jc!l!~ agad mediuDJi, 11 ~misulld [,IgaJ:" \'!Ifilh I % of em::J). I'lf dl.e c~r~lllhydr8lt-es glucasei. I~f,.llose.. maltose. and saeroae, ill. I~.hls e:x-Gllmple "lc~d Is :prm'Juce{lln both :g:lncliJStl and maltose, id.tll:tiify i!1.g rhe '1oo1!'l;~!il i[JS i\1;. m~~n!"~gmdJs. 'fhi1i: is an ~r.iI;Sensiti\<c methad be" cause ,Ntls!>1l'i'J{j i!lpp. ~j',[;li oNldillh-e. and tine IIilOOillTtlI was desi_gru:ldl. to ~t,e!l::!. rE!:r-~'!1enl'lat!.'!ire: mli::ro~~~!s:nb.~.

9 .. 7 .h:ilanUflcatiOIi'i! ,ot' NIe>I5I:5!er'la ~PJ'l'. by' API QiLII.l!lJdl'"A:iAM,~.,. AP[ O!£~cl-f0L'H ~bi(i[!;>~(!!".bC'l_n: Vlte~. J<IazelwQod. Mo,.) , 8. 2-htmr me'!~d b d..::tttrt'i:tiinil1g 'CaJi.lhohJ'~ f.il:ate me t.;!,tmli S 111 , klel'1Lifies ,\'dsilillri{~ :li1'P" tmdlM,I1Jillxlf/la m~ Ul'rr[J{j~j,r;, 'Il-SIS lnclude :l)CO[]"I}i)hyd!':~t6"C:O!1tr-o[ (C1"R].h Wl.lcn~ ~ GLII!'. maltose (MA r..tlm:toi:il' (LAC) und st1Ja,l)::;e !(SUe J, :DNase, and bem-laetnmase ~I'H.AC). f'lteTIol. red:i.s: the indicator. The b"Je;I~tion pattern :fll1.owlJi here is. dLaraf"I.c:risticor X. li~i'nijjgUitlI$. compared to N. ff"Hlorr"Jij~E '1!ii'rllii, all .flcid rcactlon in :glw::~ ,[mJy.

!SJ .. IJ lfaaMID,pl:tl'''1Il ,l'n'flv6uTS68 I!lt:n 'efiOeO'late BOHr. f~ratln~ti1'!!s ~rifl!tcll.!!aC' culonies (!lID, cliOOohul'l: awrr ~ft :srnuo[l1, round, il:i.Uirilt. naiL !)Ol'll.ewha~ Opao::JJlIl'll!. and gll.'ay~~tl, in [;OIoI~ H'mwrcwJ!illl:li rrifll.l!!'I,LZtf!r.reqIliLrus biJlh X rul~1 V fa,eOOts 'fCiI' ,~r~~rt 11: both f;;ru;;tonl :l!J,~ ~[ln.t:I!:rrJed .in 'ChUCOiIi:iile i:I.(~.'ar.

r Ir- .,1' I~ 'I~<!I' I'"
r If" .-- ' .. .~., Ii''"'
I 'I ,~ .. I~" J';
, ,
r I .... ·r IF '.- I~
14 Ir ,~ If" _.' r'

--_ .. ,' .. _- - 9-S, Idal!l'H1',lcaiUDIi1 0'1 Nel;:, .. !!!td. spp. by N;!I!I\'~'-' ,naJHB'~mupllUU'.Ei1 Id:AntJill'l,Ci!!i,tiiAn (NIHil, ,eal'd. 1~le NAI cruX! ~b~ol!.l[erlctlx VUck. H~:rx:I,wnoo, Mo. )1i,Ulli51e.<;: 1 !5 eonV~~lI,I,lmL.d iiJI:id s~I~;IHL.!b.i.Ui!llt:' ch.t'O:i'tWgenb; tests tv djJIerelul~ .:;!,~!i:!' Nd:;Nl!riu, Mt}!l'lx~j"!!lPp." and '!JIll ~ti\'CI§]( (H(I<'lr.wf.!'Ir~Wi L'I!,~}I'i~tqJfjjJW; aluli N, J'£mtv}1.rop "'I {,[t$ , Ao::iin",ba.rWwi, fmr1iatv!I'" ,!I'dm~ I, EiJ&-.rr!'Jll'l. and KjlrJ:ji!lll~ i'p~, ~ grt.m;p ·of t:!lt"gemitlms. 'R':5!t8! lndudc. ala n ine "f.l·n itm:;:mi"l ide tAt-A )1. ph~liIylplti nsphate (OPS j. prolillc-lJ-nttroat'llll,die (P~l,O!, gamm a·~lU rramyl-p:nitl'O~mil~dc (GGfL. gt,'i'ci [OJt::~iHLitro.m llide ~ G[Y I, Ijfslno-pn:iU''iJal1!iMe (LYS~, o-[Ii.troP,:lJ:iJellyl-p'iloISIJh(l[ykh,CIII.iIlC' (PHC), :~I~lcose ~GlU~, S~lI;;~~ (SUe}. maltose jMUJ, Itlcipbeny~l,ellL"<lZluli,mnl (1':1."2), r~s;a~!)r.~n UtES}. iilj:'if~iU.liu<e' (ORNl, 'l,!;!re~ (I IRH!, aIl~ ~nidiiiu G (PK"'l"~. [ncloJe (It'{J)) and catalase leAT) ~e!lW lil:'~ ~Ii';o H~ql..liH.'(l. T~oo urg~J]ib;:LTl :LD] this (ltUj~~[.ilIph i:; N, m~·~~I"!!liMi:!i.

9'..I1<1l1 Haem'DpDnus ~'f1I!:rlUi!!!inzil!l. on ,ChOCOt8it8 apr 'lel~.J.jp¥iew~_ CO!!l:inics ,nr mueeld, cm:::ap!mlat.iXI iil:D;"Wr.IIS of ,f.fcti."\!TtllJ~ttrus frtfi~rlW1L" tend to be ooll!lluem" where@s n(!1neI1C~[)SUliiIJ1;~d .strains ap;peiillt iLD1di.\rmill'IUJ', assbewn iD mJ5 1.:1:<11 m,],lle, Cof{mj~ te'"~rlw fuwre a nnmr odor; wbil!;'.b ha::: ~n d~tirib!!d as :liw.~mllJlg Iilit "dirt}' :ill~al:a:ts:, ..

9 ... " ,A,. IICJulI1IU1' DallliOn ,lid' ~,mCQ:ilbIJu. "nRueltta' by X. and, Y atrlp., TIn.' growth rNlU!lrC:r'ilCI1[S rol" x~r~"oo:li" theme (llIl~puunds ' and V-I~cti}r (I nirollnu m ide n d(!'l},tin dinueientide r"ilJ\D]. als known as lj f'tZYlitl I, ,ur Jlicaf:ilfl amlde f!{!c"nliInE' tIm w.;loolide ,[lhn.'i'Oh~tc [ " f'\D? I. alsu Jm~l'VLTfl ns COCJll zynLe 1.1 j axe im;p!lJrli:l1I1 L [ea!l~Lr,[!i:; for Idcllllif!.'J rig ,mrj~~lJ.(f1Jilrlr,rls S PO!:!, [0 this mr.::hrnl. mller paper strip or dish are impreiY-Hlbi:dl 'wil,h X ,Fad or. V lacLor. and bash IllcOOr., sUlspcl1l:sion r utwulism [S s[Jre-a, ' untu a med uun that is devoid of the fm."I.ur.l>, and the :l;ul[Js M,' I hen ~bJlpHed:l, The plate is iJ)('n~tOO ollt:rwBht ]rL 5:% CO~ a nd o11.lit'r!;\cd (ot' the presence of growih :surmlln~int.: ihe dj'blk.:s. ',rhi:o;; il'lOll'lte grows only in the peesence or both X a~d V c:tors. hid.ieared by the Iaint hilZt: ef ~mv1rnh SU!'t"Ollll1:dir ,b,:. the .XV strip, IB~ HB mOPh~JIlI<~ ,mft'~e '~llftln,g 1II111C1 n,d Sta,~/QC;:V~UB .o~rG~~ 'The bJ ad an the ~J§:;!I" poo'!!ldt's the- h~rnIn (X If.ido]']" and Ole ~t.apl:iYllOOt!ltXIJS SIJPIlUes. 'the Nlrn (V f.u':I.I.1[ I. ,j]l!ow,inl: growLb uf Ij II! HI~i:'lriLlpl;r,i_I'lI~ In ht tr:tll~ nca r the nreak.

8·112 Ild'BmirliGailc:m, 01' Nlit _ mophllul!! InfliLl~e b~ tli!l~ P.'O\!"philfi'iiili fl..t'uduretlun Tad 1[pP1']. "fills rest d ff, ·[ctUi,ate.s Nl'INtl phillJ' pam I.liflli{;ll21 I.~ ,pus] Live~ fmm l:l!Ui'm~/~i'lns IPlJJ,ntL"l'rcli'l'f! (lle~\,;I:Hive~. 1'1, !mJ!l~iiQ.l~j!,?lZw,· POSS~,;1'11 enzjme. LXlrpilllibillrlogen symh; SC. thar oon\l'CrLs &'amlnoJe- 1.'lILlilfllt:: add imn pllr]Jl1.Yrijfl~ and poephobtltnogcn, Kova~ [,B::ugenl (p-.ctilm °1l1!i'1 am illoiJem:a ldeh'Cle ~ del ects poephobiliOIlg.eI1. Porp]lynll1s produce reddish O'liof~cence i.iUt' er a \"V(IL~Js JCI!ll up j IN litiht;;ln J fi.(l nm), ~"litb Kov<lt.'il n:(;I,I!J,t'lll, H,. pallliIJjhrf'lIMt' appear::.; red UJ1'Jd H. rPlfl,tt 'fI~lle reusains l.;ulnr,I'esso. llnder 0'lI VIl(lCi.'ls; lamp, a 1J.IQI~ cuhure or H. plll'mirif/l',rtTI12df lluuresces and H, rllfl,~I"'tt~m~ lllJt':; [l;u1. illS :!ihQ'l\.11 here.

8'"113 H,g,jgiMll:l,p.ftil4:.!'. P r: inflQ\Bftill'alB' Dill Dhi!:i'ii::~ Il!Ite .D~r: • .colonies ,of .H. JHlJrtTr'tftlt~n~U' art' appf(~Xblmtrly ~. mm In diallllle'U:lf, round, somewhat opaque, OOtl gli!il£'tlilfilg. Thi!l species; is oomfu,!]lli}l' fm.l.rud In the 11[l"p0r rrop.J.raJlmj' 'ItrnCD aspart or lhG uermal Ilnra, lit I~ [mporlfll'i D ,nistirngt'li II !t frorn l.lllf'lI!fPpJ'Ji"!.j:'ij h~rl{'l!Ztle . .a pru~ab]e p,ll:hngel'l ~n th~ lower re~piram.(Jr!l' lnlcl. BfI~l1 Slx:d>e5 CElIli f.",''Juse ll!!:C1:birlru:, postpartum and ueonsral ~is. ][)lff.cf"'el1tlat1on· r the species L perfnrm~1 with the X l:uldV :str.l~ ~flg1.ire 9-11' or the l~urIJ1J'yri[lj IYrodll:ct.ion me.Sm ('Figure 9-11).

!810114, ~d.liltiinDall'DI!Ii o.r ,H. ,pBl':arJ!UlfJ.!!,~. ,Rind ,"l!;U~m~Rhfh.!~~b~phtl- _11th IMln1luk IMiIgliJ .... , The MIil11~,eli blQchemh::~1 diITerern:w.ilatlQO ~}~ii;tel1.L (BEL MlcrQbi~ olog)' S,Y~11L:1T1lJ.,; Cock:cysvme. I\'~cl,) 'LCllLl~zelllP':!l"p[;l[ disk!; W:1P'1)~= n,:!I'led 'Mrn SU!bSI1'3tes to alrli in the ,diffe£eglHaLio:n ef g;[am·,pll;§~ itll,"e anti g,l'"am-[mg.ilfi~t: jllimilOr~(lllll>lll:<:. OJI1I~embes. ::!lind ye~l"iL 1hm"€ are upp[uKiITIJd~l& 40 dl[!ilrG'msIJb~tn(tts itl;:J'U may bC"!!I:5Cd optim~DUyjrn, lhesJsl.em., Ii. 10-welJl p!Jastk'p]<lte!5 LES€d for H::ST.irtlg" III this ~ample, the foll<[t[ml1:rl;g I['Hrt![JIbytlratc~ are- tested: glJUvo:sr.: CD)", mruOO,s(! {M), s~ocrl;Rlr:: (Sll).nm~i:m~ (LJ, iJIlatlrum4 ~MNI. and x~lOS't (Xl.

,1lI1[i1ilJt w) M :S:u L MiN X

+ + + 0 n 0

" a,ll\Il,b D M S;11I !L. ~~N N.

+ + "'" + [] U

'iIlw d~Si lnll.ulsh! n~ f~tllI:e is thai H~~H1:~liUm, IiIpJ~il',tWiiilllS: liili·

1:~es laetose,

8 .. 11115 'DrOJm.,t'_ bll ot'8n:f,e:..rM' C.Q;!i!~ ,t!o:"'2~OJ'. '\-'8r-1 l.iJJiI}' ~r<Wl,=[legativt ~i!l1iiSDlatM ([om. til blood culture of ;J, flati~!!I. wu.trn 'brmx:nosis. ~(lth :lj~rd~!,llJti and ['li8t,;!m~1:n spp, III rlY ~~'DI'ire ;I, similara: morpi1ul.cgy.

9-1B BIl:t.tl!1,gHUa ,lbl"Dnilihleplfca ,~tlllS% !l!t'!! •• ,P I6lauCli ,agar. C.l)loDg~w:e ~~ (]. mill W t'IJame~er), Ilill"l, .fL1J:ill dull, ,.tu~y grow weU on 5% sheep hbxii <1~.r inctlliJa1:.ed. oliJern!g,~I;L ·.fi'b~ili !I(ll1ea:r<lllcc,m:d odor on blood !,I,_gar is ~j:rIlLIW'" 'to m.rnlnJ or 'the rtml.fertl1l'!n.H.a Ive gr,ml:l-ILCtr.ili1~fC 'iJ<lICm!, bJ!,lt '~be~ do nnt gmm 011 M!:Iei.':{lilkr.~ !:Ig::tc.RapJd UL"lll!1 hydrolYllis distID~!.Llshes t!lw :;:~i~Sl rfl)m I'Inll1~r Qther i'llictMrgmJllsl'lls.

8"1,'7 1d)""llItillil'~e:aili;ln 'CI" ;9 •. rVBmrhii' DN\I!i~h.;a!&,p~ HeM ~ urv'&i. .j3:tiwde~eUa brcmcl1Istfi,Urrt l'l:lfdroJy'zell 'tI!:'ea 1:~l_Ji(ll~ A pink ooJor has begun to (I.e''i'eJop oathe lE!'rt ~larn 15 mimJleS ollier mocltl<Jitlo.i'li. 'l'.h.e rililbt slant turned sU'O<[l,_g~Y' pi[).1I; dlwr.irng ~\ternj,gbtl m~u.I:!fiHolJ,

'D",1& C'DI!Dn~B. loti Hruc:arls' Bp. on ehaGollde' ~SlI~r- CI,m,\'if.I1CJj .Rnlrn.~JilI'~f1!' OJll choeolate ilI~lilIner4 dilJs [If IlnclIliJa[in'tl, oCfllnn I~ II6quill'e pfolonged irli!;:yhl1!~l('l!1 to ,:!d~~li!fj SLlmcr~rlt growuh 1M flll'tl'l:er blccbemlcal tes!J:ng,

!D"'20 1iI:lenutlGl'J!tlon m' g~ell~ ~p. lQr!'!ltr~ti'!1 and 1Jl1'AD • Some Rnl'C-t!tJ'ili! species hyrlrnt~~ I:.l:r-.ea.lfi 1.1:!!ili dum . . ~ 0 Ql;'lhlut:l!S .and Ii"CrI[~ce nitrntes 1!:[I !Ilimte..'l.or niliIJlJwrl. g.a'i. 'lLbd8 D51lllate Hi llrc~e-l)Ositi.ve fund redueed NQ] to NO 1. I\Jl!foi 4.8 h-OOUfS mcubaJi:ion i.n :ra]IHI.trC b.r-.Dtb. 1lUilfnrillic aJt~d end alpba~aphltft-yhnnirne 3~ added, The [pmS€1'iQe aJf llittitc is irn.die.M,e~ !Jy ill ret! color cI.~.an~lJ, [f nttratewas f!j[)t reduced 101; 'illl'aS reodlllcrn l,NII_sllIlild~1:i 10 llH]'u<n~[j 'gaS, RlI.ll: brul:l1 i'~'li1~:S colorless, A gl!J;l!Wthre--01,J [l~ a r.ing nillrOlile I)[!;]tlh 15 cU':rllinlllEd by Elddl.ilJl:!l, zlne dust wllicb reducesthe '!J[[rerlI!.IOtl!d1llltr~~~el(i! nit!:Ue and IL<IfI.I!:I~S [ilci.rndiCEL~,o1' 10 turn red, (IS s~ !l!~ilm here,

8·''11011 &0101"1'1611< 0": Sr,ueeJJB flip. ani 5'%, .• heap broad ,ngar. Thlil:!ie colnales of13mt'elifl are 48 houes o~d, 'fhe5e mi'l:'.n;lIQrgafl!isl!ll:!; llIre ~ow-g.l~\\rilJ!g;, 'blllllllQSt stratns will gmW'on I,ibm-atmy 11l1eiliOl.'i.-viII:l0J.]~ ~ddea erll:ichme"11L G:rm\'1t~~ I:ntll}l' rake 4~ hours to '~ocOOTie v~sl.bll'l amd fil~I'Ii longer fm(]J ~tlOC,]jJtili:re [mrn iii sii'l~ (_'O~[my t[) g.[oiJ.V,. CulotillBiS sre :!l1jfi;;jJ]1, rni:seil. 1JI.il1i~:e If.! cre!lllll .fin color; and glistening. Tlhe :lIi~!r plate sheuld b,e'I!Ilcnb area at '~'i"C ~ '%mo. '10% Cal for I druysl'K;.· feIN:: dj;scmdlllg :<1:S, n~'l:i.\~,

.-: -2i1 P:ailOiih;l"urelfil' m-u'lIli:I!af:da IllOIo !I!Ii<%. iliih'E!iep bh;JBdl al!!lf!ll!"~ t~ "lIillt!ci~~l is H o.mtlwh::l L r~Lo:li~ iflYi: orgaulsm that ~r'm< " siov,,'ty on j%sh~p bl~ml "gar as lJ mLl!1ri.gr,p;(yii'jI], nOl1lu:mulyLk l.ufu:my. U is oxid'·iHSf!'- and ,c;}trna5c-[ll:JlsiIH~'(' and produces iudole and emtthlne rI ~lfbo;··Ir. e.

S"e.ne:"'a~ K;ngella, ,and CapnocytoPhl'sa· pp ..

8~22! ElUAi!!ftH ,eor-l'OdiM$ l!On !S~, :Ghe~p' W!i!;g!d DO' 11". Mlcl1lscClpicaHy, £iil:tilfUil mrro,d :tJ are [i~ .. i grililll-licgab\'C bil:(.'ilili. The~'" rl:tre rastialolJS ~lliCf orE:'l~ ~~ism:!: an ~ the i r gro th is euhauced by CO ~ ami tbe. racuor; • .. he micTrmrgnni 'm grm'1o'S 011 5'.~", sheep bit! d ;lgalf ~I 'd piL"o: nr cerrodcs till' <lgi:if $urU;tJ;;e; o~h.mCJo'ed]g~ tendto ~e.u;l.·rhc eolomcs arcdlstin,guislaed by IJI:l!!lr bl~dilJ.ke ~clor.1'h,;:'~ie!'J Js OitiUdM..L.,~,iUt'l') ;m{l r.a~;;J Ilml~e-lleg..,j 1 w. 'I'h~~ nlicooof!iliilllliil] I (i'Ulll> JIlul gl'"~nv on . !~~OC(] key ;}~r <1 nd Ls I t~rlOlil- !inti ure~l-licgtl'!hi'4:. ~ I decs l.1~tl!Jl'C nilr.tj c 10 n i ~l'il~ ,ru1uPfOOlJltltlS I.!tsJ rII:.'! and urtii iJlinl:! relte· cflrbi! Ili ~rl nses,

'802·3 IIflng,iJIJD ,klngpGI'llJiln ·S% UhQe'Jji It!fam:ll BgOm.

ikmsulpic.dly. KjJlgl~rm spp. ore :pllllU[i.. gr,un-nq'!,8thrc coccobacilli <1mB ean appenr<il ~r.lm-f.iO!illlvcoocmlse ortbelrtell[lel1e), to ret,lIm erystal \-jolct_ [olollie.o; ~li"f' o~Ld<lse-pOSiljl"'(, III ud C<lhtltl'St."-n~ella] i ve, Colnnles of ;\(jllg,'lIfl' kjjjg~rt lIlre betaht'111(liyuk. and the spectes Ls: i:ndo!c- and nil1'tltt'·negllf]vti. They are faslrldim.ls U]!CTO(llJ];r"t11isllfl.: gcOVI!itn em Mac(,!)oke Ul¥IJI" is; vm:F1Llnle'_

8-,2'" Cgl'u:nieli, ,Id l!fIn,gulla ld'n,gaB Dn 11%, lid, IiMlIP b'llQiIlIdllI_'1I1i" ,.howing ~":IiI'!H!I!I!dI".I!.~ This :Ei.~IlII]je :shm'llll 'the VC'ry dlsnnet 'bc1,0I :Illmlo.ly.si!ll di:i:J.lla}"I3d '~n!TI 'llie ;agaJr l?~aJlc ll:i-l~k].1J~ LQ Ii li!i:ht source,

9-25 'CV:lonWe9i ,of' ,G&:p!!IIIJ.CJo'iIm,pJ'ila§,o!II DII'II 5%, Ut\eJ;!IP blDc;d ,HgHl'i" 'f~£j' ~I rs rtliilIJiuus, l3t'Llltathre runael"OLleS a nd ma_".' f1l1!N%l1r i1::l liny yil!1'lJow ,!::(ljoJ]i~ after rn't'rm[tgl1t gt[~Wl~1 [ill bkK!d,~~t; Growth i:l;, enhanced by cn;!_ ('Cllon~c:o nfol' iiill)w~r(!wil1g and {lJ'Llll'rI ,~~IJ~irE .il, g hours 100 detect d I.~tln ct ,t:o~D:llil!tl. i\1i.tr«irop1tully. CilpIIOC!l/l1jl'haym !ilf:lll. aJre thln, gmi1fl.~ 1](;;"g3i.il,'e' baeilll wfifh poillted ends, These m1rJrno]'gfl.n~m\ll '<ve:r'!ll prcv.rousl~r d\:BssiHed m;llt1cUtl'lllidl'S' S~~.R rtd as CDC il'OllP DI~-] ,

8~2Gi ,e.IIJpn'DIC!~,h.g.I!II O'nl ehaeD~mta ,aga!:'" GftlJ,lIill~l 'rln chncelate llig;;nt Sl1flilll,;S the effect of ,glll~]lll:§ m(JHllty on lll'lecrllrm,!a'l p!"m:lHceilhytlu!:.~<! 1il11l.':milr8;]rl1l~.[lfi'l$, A. Rim Sl'bi'· r'illll1ntl~:I1g '[ he cnion tes is dHil;acter'l:l!1c fenr ~r01i\.'th of ~his IMi;rn'O(lrgall!Stu, Th~c' rn~;crQO!'!l;am'S!l\3 do not gl'(IIW Oil !\.tacCDPkc), agal'm'ld nrc catelase-, ooi&se-, :tIflOO~~-. and urea- 11 c:gativc.

Os'mpVlofjacter andl Vi&r,iG S,Pp. ,curved gram, .. lneigailive bacillii,]

l.. ....

~9"l27 GIi\i!lm :l!!I,tAII!iI of ,C'.!!Imp:yl,li!lHIemJli· BPip. ,( >0: 1I12!!5DJI. MicroscopIc Ell ~y. tll'mI'llljJ~llI(ulf !:[lP'_ ,:1,[11)001" ns t"lLJe'lredi. oomm,a.sn:l:lJJed. S.~h';Jp.eil" and gl]n·~~"I]1jg'ed gr,i;!JWi-neg.'1lil,'C' h:IOl1lt '[h:e~ ~11:'pes resu It £r(.1l1!l 111<1113 cells re!Th,E~llin~ ,;'lU8:ehcd after dl!i!!'lsl,'U'I _


9 ·ee, Idenmi.;::aiUOJi!~.f ,CBmp~ellil" JfjJJJJft~ b, r'i'pl'lf:ii:w: Ie iPel>d1 :sY'lIee,ptlbil[I~:, "fht.i.$ microoIgar~i$m: ~r(i't\l5: best <It 4l"'C. but will not gJn:JlI': at .H"C. It is. mi('rIjHl;ro:p,~wL<:', reqltitmg 3% [(I J5% A.! ,I!llld.l% to 5";l' .. CO;l:' SIlo Icc:~~,'I'C mcdla [Lc .. CIUlIJYhloM agar) ~~ .l'eqll.l:5red te b'(!lt.I:t~ rftiS argaD1i~rfi frollii mixed ftlora. It les oxidase- l1nd llilTIUIi:'"~iSltive and U[i~1t.·n:eIDli.~'!-'e. l!. di[s<tin~uJsb illlil ci1M'acl:Cr.i:cstic ls its S!LIsceptibiJ:Uly 10 Qali(lllollc acid UQf~' ,fisk) .;;w d r~:S.i::;Liill(:e Lv oephlllot!hilJ! (lwtlrnn ajsk)~

a..;2aCam.p,yI;~l;Iitel'" J'9JuHl On 1'51 uli!!ntwltil !QiDld ocu,t;pg, 81m1ji!iI. ~bS is deleclcd wiCi'g 1t:"<ii.dI ~ClJl:dW paper, butnot leu the 1':-;1 medllilm. CtN'1¥,y1!Jil'n1tH'!'" MwJ..i Ellliuhtdr{lI!J,'ze:5 hi Pl?u1w!,e (PlglJl"C €l-"14 on ,~, 40 J,


9"'3D .~ 'Wbr1 co ,c:-bo,ht,i"'.aa i~n~% ,.'hee,. bl'oadl lIIIg)al'i1l Colonies, Oif Vihi'lo dwkmi:' ,gil''!:)\\, w ,tI en moot routine 'n~loornlory llli:"!:Iia, Grm,,,ftth i:s e-.!1h:anced by aoiUlioll or 1% N.aCI. Vibrio dwkm(! tolerates 3% .f~CI. la tJxidl:1 '"-positi!le. O~,~, (8.=.'] 2),111- dole- .. ,lys~ue-- and W:lli:l~lllJR-PO:Sili,,'e, 8" 'lWbrta ·i;'Itqf'~"",B ,~" t!!l!OuulfmD,.,G trm.-'bU sa'ltill".ur,;roH ~TCB,!S'I a;gDl"o ~oLoni~ iJ 1il!)rIv l'll.rNetm:' ,fJPlJl'ur yellmv due tn suernse fermentntton ,;;;0 TI':BS. Abt other g am~Mgaii¥J<; billl.:mi are j flh.lb~tc,l



~ -31 A_, WbFtu pW';&'Iiii&en:ra,f;yUauII an ,5'% . 'hi!!iB;p 1~lgod ,q!l!lll". Vfbil1ia P'l1'cdia:il'nlO1!JHI'~j,~ dees I:\~ubr,~ Nan for grm.~·itb and cam 'lolcnuB1u N~LCi. (olOl1iie.~ om 5% sheep' blood Q~ar a rc al)pnrx:iJu-Li:lld,~,' 1 to lllUllll ln di~Hneler, chall!, and can II - • I,:i me a p<llc blliC~g[iO:::D color, Colonies glio\vweH oJ(a;cGonkc-; ·a~'IIi:, <JHO: indole"" oxidase- .. lywiI1c-. ,and ol'D.lthiJn~ d'l;!c:tLrboxylase. ptliiliti.vc. 'fIljS pecles r~::nments glueese, maltose, and! maunltel, Th i s or ianl "Cn has 11 ee!ll assocrated wi th wound] hnrec'tlou8nd g."I~ lroe:ntmH.s:.. II" 1Iiirriv lO!!iI'ralu::rJiiimulplc,U8 f'!l"" n eur.CiiinlS_J! ,and \':" elrG~ii'ise ()"B'llGw GDhliililltBS, onl ifCgS~ l~ .p.i'litJ1J'II:'I'M.()'J!ilkj~~S d(.1!!!S l1Iot. ferull€llIt Mlcn;me; thus Crl!ilfl I es ,r:J1!"C green,

Nonfe.,menla,tiv,e ,Gram .. Negat.ive


'9~32 ,g~Udat_tV~'·_l!'mll;!:ntatllve ~O:F') IIHedlllm. Btl~I,ei"i8 lncerp nue cElirlmh"dra~~ bJj' both oxldailve ilod fer", mentarive pat'h'\~oI:l!!'S. as dc~(' tediln I IP m.~dium !Hu8h <:Iud ].em<icil1'sl \'Irlt'll oromLbymcnl blue p'H I' dlcator, The 1,,;\>\: pro'" ~dn-fn-cDl'ihol1ydnlile [3Liu ill or 'ulwtli<l pJ;'CVCBlS tifle' l1euJllml~tlID (;If \!,;'Cak nc~d by alhlll!]c t:'ru:II-,PrOOl ts, The rnl roerg\..'mi I d_ 'sCl'i'boo ln rhis section, i:lre mdtiElLh' Hod li1iJ1IJ:OKid~lli'tc ilfOlI'l'H'ICgrul"h.1'C baiCfiHl_ 'The h.d~ uu Lime ldl C()I1- tains HI£ l~iI5i:1il rued um ~1(.m e: the ether 1t\',10 Lubes contain dexU'O!re. A, pn 1ti!;c '~est (ON'id~th.'el ~s mdtcated by a Jocllo "o~or ~flm1~r tt!lJto) and oat :ne~aUve test ~ncmoxtdaUn:, DOJlfC'rrncm atlve] mows uu (jUlu,£" change ~ gll:~ffi. rjgfll fijjjl.t'l. Fm:ru.e[l· urH, " fllll.'i!·uf[!.alllisms pruduce ",'bid ill oolhhe ox[da[[~'t" IUIJC[I) tubl' and thO' f~r~lc.rD1ative tube 1[JI\'1IT,~.lid wilh mil:)!er,al oil. Flgur~ 5--6 on [I! , Hi.

'9~.:33 A;1-a'Udgm:anDS, nJSlnGniB un 5% .h " p' IbIOQ" .gm-. Th ealornes IJf P.l.tlfil,I7'IO&11 on 'blood agar', re typit:311y Ylilluw-~["CI~11 and beta-h emlllylrc. [11.uore5!."CiIl Iligmmll'j;s, ace prmhll::NI by some species. bu~ (J)'Ot'YlLi:l i 11 (I' III c-grcen) [)t~· [I,l III L is pend uced b!{ f, (:rl1'rl~'!7"iOSil IIlcme. Mw,L cwulli,~ hiJi'lre a rlfis:lij:flrl" gra pcllkc OO(l!I" clue 1,(10 mnilrlo<t'Cc:!.opli3raclI'&C. r~ c!rr,1 !It!~I)'I'~ lay 1'1(' (ll.stHllgmllish d (rom other spec'ie:; Ily it! ahil.il,Y ro grew at .2 . Hil P •• I:Id'O'lnOIIJIiI.1 ,SB.i"'VBffljIlliAOi an ti%, --h liP bJood~pr:. Ohtiqueli ~hUillg, demons ates m~Lallic sheen un ~hc sW'lue of colonies.

9",9" ~~dGD!!O'na:!!i!l ,~r,u9'j"'n!~ ,!:In! Mao .. ,c.g'!'I'~"!ii9~,".H"hn, b~~!c~~r,~'Cn eelontcs W~11Ji Vl~.}' lii!llm,Cl rC,11~lIcrct1 c.d~t:s g'r(ll!iiriIng oa !\}JilteColhh:y i_~gOC Colonies grow W"ell~n J 5"'C aerobicalb'. TllCr 'I\~ln a[tS'og[\U\li,U 2 s-c,

9,,30 P...udom~.t.!IJ!IiN"ughtctBa 'on 'TSII .ral!llt~ OIJi~ J~Sl agjal(' slan tj~S~lld(m!f!'J':ril:!i' ,1"I!!l'll::J,rr!(!St'I s,Ppeli[\'i" 1I!l:3. blue~eetl. SQme~!;'h~t l:nel<l!~k', fll.lOroe:l'(;e.n:ll;t'l;y~t" Q[ gI:'(lI'Wtb,

9 .. 3U5i Pi5lmllll',f,I,1 pn:n:lllilcrti'UliI b,. p~gd'nrro.nas! 8!1i1\n1!S'ftCliBa. 'f\'Ii'O stx3i.lIlS of 1~",!!~lmnm!r;r;;, ,111'(u!]hw.mgrul'lfi!L~g on Mao::('ornk"e~! agar, iE,he red pi91Ut:i(iIL, it! d~l!;'l to ll.L{j' pruduo[ion uf l)_\IOm hr,~n. 3 noufluorescent, 'I'.'alef'-Sillublc pJ,Iil,l[l;(!][i[, and the yellrIlw"g[8oo OrllOF is due to ,~J'{~~,'mlilJlJ, <l fluorust'lCrlt, walt"D'-.5l(llubJ'-e' p!gJJHmt

9 .. 37' ICl:iIIHllne..l!Ul'la,n of .. eua~!7Iun~'.ij, ,;!liien.~ ,HlIi&Dby t:il:liilllt'aIC,Mrila.li:l:;i!lI iiii ONF' tgbe., The NIP S}';S~ I!'ell] ,~n~IfI]JCu 'L:!lXI:1;ll!,.lJI"ile.Sl, ~ enexa, K:a~, I 'IS 'UI~dI' fot fhc~~l!!Iljj I!i. ,::~illilll {Dr g_ratl'fl-{F!cg,;,'U'!i'C baclm ~bat !'.IOi1ot bel:ong to the f'i~(,I!'l,~ar..r."litrr.rl'1ll_ The .s:ySi;cm COl'l s...ii5ls, oj] tile GNP [g:lIlCQSI.". r.;2~[lIl~I',~ceLn) lube. Ih~ 4lP !t'l1b~ jHgur.-c 9'· 3 !:I.) and tile tlnlNIP-Tck I)i~~c (Figure 9-39),,, This ulliCf'OOI:'J.!$!:nli~ producesa I1:llJOfiC'Ilcmg IP'Igta;ilBllIt (u.mtJer.[l Woods ~1UlfQiPl. 't~e red iHtlll u.1JIa;sr the collsLYic1io:!JJindicaL~ N.! gal> gooducti(!ll:l, R ,rJttrm'!:1itmSij d.OL"S nClUe;l-m.eillgIIllOO~.


~.".F' L-,o. ~ f'!II"I ilJ71:'1i~·~


"'-"'" ....

, ..


'8,,3B fde~UIilc.i!!tl:g!'l ,011' ~~.m~iI!Iailj! a~S'i'" ,ftD5!I5I' by UNI;; 'Urea" '!Ii!Ind P5'g,u;d'C'5urn (42P~ agar. In til u; [~gl.!n; lt~of; GN[~ $lJiillL dcscrjbcd iu ,ii~l:llIJ.re 9- 37 is oli:l the l,iJl.. iJlprn;i1h"t! m:lia sln~\t {fjgIJPL:- 8:-1 Go O~ p., 611 ,IS in the cenrec ;,md42P. l~.l:i!lrondtubi:lo[ li.:w:N/r 1SC[(w1l, ls:o.u.lllH::rri!ilhtf~m 42P m~diu.m, also kJ](I\\'1] ;~ preu£iooel. "gar, is ~1J.{:Ub'~l£.l;1 .fI L 4,l"C for .I '@ 1;0 11 hours, ~rhi}; n]t[[o[lrw1:I1U~ gret\- ,at 41."C and pr!;lI;lttceil the bJLle~gr>ee'J1 Pyu[':Y<llni.!'l pigullJEmt .BIDied 0.1] the ll,"Ou.:Urms l~~ t Ilis rigure, u he mi rmmg<lll i!1'J:I1 can OB Oe'Ii.nitlwrly M(:nll:hk:cl.af> P,~"'IJ[I.J)mr.mru,~ {l'f"I"l!gln.rl.'ltf,

9-4!p Pw8u.dl:iomOifJiI!!iS _Ui~ i!:I:fi! il!lio% !l!lilh!iI!i!O!!!P itllilliO;d agar. ChSJmcreTi~t1lC colonies ()f l'sof!!dti'mOrnl:r S-lliCet-J Oil 5% shee,p blood ~lg;:lIT'are burf,.co'fmfli, dq', wf.[l'Ikll.'d. ami! ,u.lhcre to the (I~a-r surface, IlrH'Ik~llK it djffu:::~tH [0 rem.a1.1"eII1.ern rmJ'i'lII'le plate. Most dr,",1n:'l grow' at 4Z''{' :lnd reduce lli'OI'all~_<: to l1iIJ;u~x'l] r,.I<I.ti.

8,,3B IderrliHle.aii·lolil or .PSUUdomlH!'.a!l5!l1 a~!f" 1!'\!!liI1!\!!O!' by U!!1Iii·NlF-Nk tl;:!ig'bil. '['he L'IIi-Njf,'-'['ci-:plii!&c r EWitw:1 Li:lbOjj~tmi~s. L~'I1':t:o.; ~, Ka[l.) ~ if 1 ] ~e:,t unit !"'utl:sii>til1~ I~f H1S. Imh:i!le_ ~ c<J'JilohydraJle cmUFol.~rIJ.,lOose, x~r~i]~, man~ i [01. laetosc, !ll1!l~IOOe, acctamlde. CSt~!J[JIl, urea, [)Nf~5C. ami ONPG frigttl"e Sr]l em p.. 59), leiilC'lilJotlS fur Pset.rilfl'mmlIl'S m:TU_rtirJillil!ll!Lre:

G~,U ~f.

+ fiSC u

M"I\l\' !lie M:AIL
+ + n
1lN!3:5lt' ON.rG lillJ;S
+ + IJ o ~~"fl o



~h!l'm ,Sp"lli1,g~_mJ!iil!iiEliJ!!Il t_mlI:Ii!i'r;I}f .NeU8pjft.g,.I\t~}' p~;a.lc;~.tii'l.l~laliii ,5'M;;l!liheep WiD~dI:ljr8l1l<1i'. Collm:i les of ~~J:lrungat~1 i'JIM)~ ~lll.'llm~~Ut~ lln ~% shcCIl bl(l!odi a~r arc smnrl nnd :y'CUm~)bp'j.lMn':llI!Cd a;[l:cr 24 h(iltll'S Cij lru:~~batirOn, Tilis specieswas :w:rc~rlot:!s~y known as P5'e~ltl(!m<i'ml:1' p.!IIll('im{l'iHU:s and CDC !!llfO~~p 11k = l. ,It is ,81 s~o,,\o..g.m\\iwg m.icroor:,ganism ~l H"C, bllt oplliIlfl.a1 MI'OWlb OOC!Ur3 801 30~C'. Thss miernoegaJ.1i1l1n does not grm'ii: ilIt 42 (Ie or OIl ,M1tCCOIlk-e] agar, his oztdase-, esC'l.Drill-, ONf'oGa. :lInd IJN~lie~itj,v.e and nill'2!~eand ind(llle·negallive.

.--4~ ,sw1il'PtN!Phmmolla. eXSnlhoma.nI!!i!:!!!!!J mal· f~"mB on ,5% .bil!i>i!iP ~WQlOClI ,Bg;m'o ~"o~omles of SlsmJ.lJ'ijIj~JO.llUfflll~' ,rn:i'J'lwphjJ.t,~, 0:11 !!ii% j,l1ec:p 'hllD!)d ~:ar <tr'll' chartreuse 'to 1000~'1;.!nd~J] a.nd lla\te a,cb&liracleni uc SiJ"Ollg, uum F.I1IOl1i~ OOM. I fili~i1: other ps-eud0J11IJI!larn1. ilill!i specl:es, Js '001:'Idao;:e-negali\l'c" lit is f~_5Qstallt to rrumy 311lil!l1acroblal. 01- tfl(!!t1rgb U'w{1iI~(ljlriiU"'iS.uIfam:illl10Ka2ule is U8lJ OIllJlleIT6'U·~jw,

:Sii .... -44 BIII\8wal'MNtII' ~pae,uaama."'\i!U!lJ 1I"'fR'!liH" '~~n. 'Dn !5% tlbqp b~gH ii_F. Sfit'wandllll51'lnen,cw geuns Iifli:urre IID[ the !'!l!cm[lol~.lulli!lifi iJlm"'~O'IJ-5:]y d~I~1fllj}d as F~,f,fdbmoml's rn1~!~ ,fef S and ,CDCgr-Olllp lh_ Co'lloni'CS of She'!i'!JmeoUa i!W~~r~bi!)ns (10]] ;~, aheep tilm.li!l ~ga[ are :!!Ug!TI:!l!y1"isCUll! and IOU: D!d<md plnkis!1 m r-e[I.hrnv,m 0 (ll'im~tan in euler;

'S .. 43; •• "ItlftCBttDn ,&1" ,5feJ10tr0pf'lgmg.nlJ-l5!I1 tlt: nCf!i~mO-.i'Uf"!' nmltDJilftfli'B tQf th., UIi'II,·,NIF·Tek~ REladjiIJn:'i roc 'telwtrtwJr,m,mf,l~\.'I's mJlO!~r1111!i1 arc:

GlLiJ XVI. ~;tlt' LAC MI.!-\JL

wemlk 0. 0 0 +

sse lfr.-eiIJ DNase' ONPGH1

+ + + [l

8-4:1 1'BfI Ii"lIaiCIttlanl 'm' ,SIr . waft a flal IfPlAIgda-. 1I1\OIt~:J 9~"eli:l_p'_: '11~"k~ dLHI!'a'Cl:erI.8tl~ dml:diIfu· ellJlj,ales iJi~t'lldarm.!'J'llijl Jmt~(.lms :rrom f:ro::gumltly cneountered mon[en:neif.l:teT.s ill1d ,OIlier related rnlo1oorgfmiSiJ1kS istrs abilil;y ~u iJ'fudu ce ~m~(;: M1titLI1ils of 11' S [l'I 'lSI 0liKL-\ sLrnnts.

Other' N,onfermentatlve aalc'illii

'8.... tUII.;;I:GIit aga.... PmdlIdiun 'llir "he I:"JJ:,:.'Il[il::'[Jr.:;'i~, which bydrfll.\l:li~ IYNA, ID1i!Y be t~::l6I;1I'!J! tiiJT~[(;il~i a te nl"infe'l~· lllCrllUniij B,r~lil~-l:'leg:Jti~"c bacl>eri a as 'lh'1:!1I <I~ S,rnpJJ,!,tJ«CJcnLli ,PI~,4· ret,,, aim! S'l~i'r:rlild' F,lkiP":I1',\WflS'. 'fhjll D:Na!>e 'l~ll1lt!diI;lLU !;:ol'lll<~iml OOhfidi~lt bl~taml1,pl.cxl'(i wilJ! bNA., Hy(lm~~!-ih;,[')r I)[',;,;~ by the LrJ.OCtli!llml :tlllcwor;ganisr:u cml S1"~ dl11f1 ge.-; Dr struelare IlJf Lh!;l dye to yi~1d ,aw :pil'lJi lulur. 'TIJ!]ljJldill:c' blue [1:1:.11 i11 hihU g.nj)~'I'1 rn or m'llll' lI1irnmri§:lllisDII.:l~ w ~ILJi\'Ott'll results shoul rl he Tetm:1ec wi! h mmnh":::f method.

~!I!r:.~7' AC'.tillw,vbart;:'ki\r' If~fjJ..,tob.c;t.J'" I~'" ~'a'QfJUCi!.;Ui: vaT. ,1Ni!.:ltra:t~'1 bBiim'iililiinrJ' ran Mae'CUifilliIior 3510:11"0 Ar~j~~i,QIJctrl,ltr r:qmm~mm ''''1i:i! rOfmedy dr,S.ig~ n~1!I ed as A!.'.!'ldDW1dt'f Lnk[!t[(i:'~id~l 'l)"all:". {I'Ilirmimi, llIl'ow:rt'lhr hlflj1Ji ~Ii-fjflt'~] L~lyJpik'EJl~l. !md M'''!d l't,.r!:l"IOrj~'~f,! .. They do !.tol [lr{l{lure a [lj:~l'Ilenl mll blood ~gart, [)'!J[ appear rainl Ilirl.k on MacConkey ~g~r> [J,,~, sI1Ll!W[I. in UUS Ii.tt;tlre. Tl'ley 8L'r, ffi:idm:~· und fUil"l!r1e-l1c,~njvc. Mir::roscupi'CilUy, tbese ,m.lcroo1'J!.c"lfl:is.:ms ilPVCEiI!i" Ol[S ~fI{:(:nbm::mi. prOO[lllllmmUji i [J, .~,!.ll'i). ,~~l(l fDI!" Ihi::: rC<lwll ~ I te~, tl<li\:.e heen i,J[buii!JJ;s{m 'ii·r;tt.fi'il Nl"is~·~J·iQ. and MilJtlXOilM1.i :spp.

9~a ,Ra~il:!tlacferlt!'m! ,m:~I"D.-.pifJC!Um 'On ~% !aliii!eep lbl!oad i(!!I'I:!!f;'. Colo:niif!> of flm'Dbnr[(JiwH ! Ilrll~il'tg.:m'rllic!l1.rl em S% shil3t:iJ' IJl.ood ,a~@.r ;Lite!!:' "a ~1:JI;r-'S o'J. ineubauen 'H>: [I ppro..or,:.i DUd 1£131 1, 1111]1 III cllaj[le-te~, Growll] GJI] Mat~Ctl!lkl),.'Y ~lg;.r is nri able. ,HlI1Id rnoM !ltralns aee l»!id~~·IPGSiIhn:. Thls ~1:Dl"r]es 'is lndole-. 1~M;t]lia-. ONP'G-.illlJ ])~ a~"pl')si· thtl::. 'ilIJ'l!a-IliCgolnw. ,I rid nrm.!lwliw.

'9 .. ,;119 BCiI\ .... liDl.sirlBj' (Pt!lieuiUI.mdMlIiSJ ,~e.pa.c:l'a· on MlE!!¢COniker ~"'9l~r. TiI.1i!l speeles, fo:r!IU8rly kl.t!lJw.~ aJi~ fi$f!,l(WJ~iOrtt.l:l: t\(~).rIC.r{!, g~ :>'IMIo'ly. c:..'~;peci=I!~y\\i'.he:[J reoorered fr!l!n~ cy:mc ffbrosts 'p<IiUcnt~, In WhUllU ilt ls ;) s1gTlii~fi.ca!lD. p"lboQ;~oo. ('btomes on Mtl.t1COll key iEl,g<,r <lire otmm IJinright ph.~k erred aftcrp:~m]~d iI~l';Lj_lyjUion (r~s. :;hfl1l'Ll1l"l here) ihl:i!·tJel lactose CTh':i:tilall!jon. '[lie ~ic;sitiI.O]Iit'l!rulil'1'0,gil.I"'c. Mll:ioough mRl'!I.j1 stl:<Jlins. d!ilSPtlll!~' ".. weak oNidiaro rEHd!i],.'it~f, .Most ~ltams ate l)i''' stne dii:C'a.ltoo.'lyhu:e·~~!,I\'\~md ,O'.!,\jdize ;(I. ]lum'bcr of ru~<IrS. i~ Id,.dlngl actose,

9 .. 15.0 iLsgl,Q,litPM,a pno\umo.pbJIs Dill b,u.tIi~ !I;!ft"'j'~g~I·JiiiiQAt ,iiIUtftaC'li: tHeft), aglllll':. ~C¥1t: alar is; <I ~Iefl i'l"C med ~~I m f()r the T~COJ.!eryQf LtWiam"ln ,spp. Thls bUITCNd l'tloolllJllfl ls the '!;g<JIf Ql' ~hujm [Or the i:mlatLU[1l o~ ,[.eIJJ'ulielJnSP[), because it coma ~I1S· the requsrernents [or o(lI.iJ;ns t growth ·(:If i ha mleruorganlsm: L-c:t'Sli~llle. i['(l(ll salts, 0lJ[!(l.·~ pH -Dr 'D.9" .M~lIMOIlcs ~ addi;J 1.0 ttl! !Ilhll Ilheg,oowlh of other bucltrirl, GrtfWlh ~ppcarf,: hI 2: 10' ~ d~IYlJ., and Ul;e'·cr,I,1ouies are C:LTCI] I Hr, ~l:!lh:ming, ~l]lllre.. and mCilS!UC U pro 4 mm, ;:J:!O si]Q;~~'J] heee,

\ --

CflAPTER 10' ,A,na,er;obic Bacteria

naerobic baeterla are' unable 'lomu~Upilljr tn thepresenee of atmospheric oxygen. although they bavt':varyiIlll degrees of mygen toleran .. ce .. 'Ihe ,Pi~lb[lgemc: strainsare dEl/furled into two groups: exogenous pathogens, prjni1atily the spore-form~ng clostrtdla that live rn, the soiLL and. contammate wounds or spotulate ln londs to ,C~'U;Q::';; p' disease, ,1:1111 d! the endogenous pathogens, normal lnhabitants of human mucosa. or lumens that can se disease wben they enter a l.1armCliUy sterile site alter traumatic breakdcwn o.f the Dormailll mucosallntegrltv,

Specanens mus~ be protected from ,H1r durtng eelleeticn ,transport., ~d ~,r.:ubClrator]l hall)dlIlling to maximize iriccmrc'ty of these .f~:&tidi.oli.l!~ miCt-Ot'lr:~il~WSr1]S, Because of ~he time and resources necessa ry to, ,perforrn good anae rtlbiilr' micro b~o]ogy,. most laboratories lii'nLt ai!1aemb~c ]!:mces;-'l;lng to specimens like~y~o harbor anaerobes, such. as shscess aspleates, deep tissue biepsles. and specimens ,collecled! thmugh H protected catheter ~st6:1l1.

1:0,,1 ,cCI!h:l'.n'lttll! a'! B\aG~. '~lfjl.:!i' 0"8",,'" Gella bl'otHi .;apr:.. &d~roid;;"S SPJ:).. M i;,\rclI as Mhor. anaerobit; ~ra!l[l1-ul!gatirve badl11 llfrow bM OIl. tim: !]oosl;ll.ectiv,e medi 1J~]j. AgaJr DilJilS'l. Rl~ :sD.lIPiP.letllLi'llI.1,Bd.\vith h!:l'IlliiD '.l:I'lid vUamLI:l Kl to Sl~ppo.rt the ~Jr(lfl;.\rlh of!1ll<lIl1.Y S"~ie..l;j of ~lTIIaBmbBS. 'Y.'I:i.mm loo~(llj]'iJ8:'l £iI~peHr as clrcwar, entire .• mdralwd"


1'0.,2 'Colonl.lI, cd' Sl!IchlJ'Dld8:11 'I'Rlall11l!' illH'li B'BCl" :kiinllcte.u bile 'rill!Je!iiiIlJliJ ~ElBEJ ~iAli'i. 'fhl!l rlllooiil m COJ.l~ rains gen:r,Mni.du to Lfi.IJJibH: f~It]lltMI'l.rc ~rilm-l1ea:IDIIJ'vc h"cllll, BjJJe· :!iOilJtS il'!bloi!t II.J~~I al"l'ae!!o'be!l PUler :l'harn th,Q~!,l' of the .I!t !!TWiils ~WIJ.Jl, \",bich ~ as ~ar,i}t .. grayro b~.ad:: colonles due ~!'l' their lli.J;\dHI~Y:si:s 'of eS!:l!1lkdll':l.I.!L1f! .IUedi'lJlLll.

10-3, OIl"O'W!tIll 0'11' ,c!loall"ldlal an ,Bggi yQII~. ,aRiDl!". The J!md~'Jclicm Qr ~he eF.l't!{I1n.es ~ccHJilII'Hlse. o~ .. que whlJle pr~ dplUrtt' cxt!!fi(:i~b1I!ll (rom the coio!]:y W:W the medlum ! rlfjJu sirlt), alii . .;) i i;~mse, [ri[JeSvell'JI: Sl1.{'eCll, (II] S,ull'iEilc.t: off (I(J~imy Utlfl ~~idl') is: used ~o d'iiffer-,enliate OIJllll(!lJl]g C1Q_'irl"iruiml :5pe('tie~ and to hetp idelJ!tif~ iro~e j; U:;OO!'!~!iC'fJ! IJ It :species:,

'10'·,5 spot: (ndalle elilsk; tiU!.t:. 'Il;)I~c.str'Orp[(~nC'Cor the ensyme lrypwpl1ana~, I!l plalr!. (illl".[ p1ipe['di~k:isiplacc::l OIl !!iLl dl'C'(t of g"[,O·~'IIth. or .O!J rnl{:rQOrga rnGm 0111 ;B!},l.eclhaID CO'llta~l1i[]g L.:L'"Y[J~(lllhal1 tsueh as most 1)1000\1 aj:lM ba.:sesJ.After the di!sk has b~mmt liLllli:sl j 5 [~]inlJit('i> i. ~L d:wp 'ur Jl'CIfiIIdLn:u~,tL~l;;;WiliI(t:.:iH- 1Ji~11i'13.1d~h~rc:lfi 11,;. ~l:iln:d on ~lu; disk. A grL.:(;'nii>,h~I:!llD;:: {'oIm: il1di.~ catesthe IJl:eseHce nif illdDI~, 11 b:li'eakrlO1i~~I:1. pmdilld Df ~'rJi'!Jh»r ~hl1n. :llmIolc i:<; 11:;;00 to help dil1lel'enti:l~e ~I!ll;cmg ro"H~:

Fjl~ti~at~l·irlum. P~f(ll'l!bliflerrr~!!l'. l-"'!j:JI~!JI.rmR:Qn.ri8'. IJm'lllelif!. i!lilld f1eP'lOSlijl'P![)l:ot't"~I~ ::![]p"

'I,D-A., NltJtlori., disk. ~.IiI$i:. ,[bl~t;t. far rurJl.]~-~j'~n QJ !lL~r",11;! tn njhi~e" a. Ll"ul.FdJte~Hnlpre!;ll]al:ed JU~.' paper di sk lis pb:w,ed I;!!1.W the ~!rn('.!c~.11 LI I'll or ,:;1111 ~m~cm~i(' ~lIb::uli~llfe (l;r t he 1!.liCHlfIT~8if1fi~mJ" After 4:3 heurs illlC'Lllm!:irm. draps Dr the llfitr:Ji~:el"eagellll'i:. SU ~r.mJllc :;I rid and alph.a-Ilaphl hiylamtnt'. arc added to the clioSk. A n.:d "olor ~1LI.(] tcares the .~Il:'I(:SCJ:1.1:\C' of ri.~mlc"

11Q .. 6 S!!:!d!lj!!ml ~1~~:!"!.~tlil~II~!'!llflll!n§It~' ~sps~ I!';!~~ :t,last'. f'tl.!M:,iw·lt~,~tIl"OCl'IIr> mltter"JjJ~lr Is the I)nl~! riIIl1Ja~,rol't~ gra:nlTpo.<ti tU\'e !:m:cIJ:SlI.hIlU i:s i01ilibu!edby SPS.

1'1;:1 .. 7 !S,pee:I:aJ IIit-anGY anllnllcn!:blal dlglw 'roll' I3AlIIUmptk'!iI hI:I\liIIilUrhi .. sUDiIiI 'al ,SDII;Uiit.lrDlilli!lI. 1lm paUtlnl, [If ,~I.I£~-epI.fIlUity ~tl ~J~ei lhN:!t'I <llIJ~jbtWilk: disk!;; kaniit~·t:ill, uoo I) ~ll', 'ouli:slfurl. {1.0 p~}. and, 1.iilnlc(!1Dydn I~ s Ill:!!:) ,(iilIll I:aelp dil'IT~H~Jl~'li:llie d'LTaml:8 anaeroblc ~me[,m1 1:!I.~h~g Khese (l[iliEir.ttl;


1mrlt'rG!lit's j:r_ gI'Mp .Bacl!l'i;@J'd~s t~!to!Hfici!;l:


.[·~jro~a!.riimi sp~i-e~ Vdltmre.II!01I :S'peci~ 1~'I'J,.Iwr~.i'rtlaj!l1S' specit{'; Pr61iDt.(Ik~ specj,e:s Pl"{lW1>! ""p!:ocomls


Oll1oer gr,'TIJ!I].1](J08ill~e eoccl .s: S

~ I) Sffm~ &~~n8 are kalll(!lI1f'Il'Ii!tl-rciIl~t.m1f1 ~ 2J V:. "J;aJl'~a.hhB: R. I:'e:slsl t!TIt: 8:" ~!1l:sre:p[,lbt£! ~ 3: J Rtim ~nLiDl~ rulH;I!II!iJ.:e,p:ri'ble

SU.l). J1(1}



5 ~
B:. It
.~ [JI s "IO.BA, S!BehJn::illdO. :fnlsflJ'. Oli'! Bms~lg blood .... Ba~. The [arg.c g-ray. IDil00.irl eolnnies lind Ir~Sist8uce to alltheee Jll:!t~fIICY dmks ~!iGtjhMill~igll!re JII).7 ar-,e lyplCaJ 00 lIrIieflilbefs iIlff [he .1l fmfJJJ.~ group. The 6;.,;'tr1li 'illter :p~r (Ili1';k is for pe~]l;:II:tce (I( t~,c ruil:ral:,e:te,~t. B:, 'QiI"l!IlfI!l ,.t,i(lfn G' ,EI'''C!ite~ld8. ,"" flN.,.. ]l8i1~""ih:l~j]llll!!!i~ ~l!:rnm·f].ef!!E!:ti'!o1le bac:tlli ,nth rounded e[lds;"Bmh IlIe(]l'llllj)rph~N iL1IIUj in;ugulm'iily in 8ttai:liti~~ can be o~8rved ill dlis smear (X 1 ]50~,

1,CiI'·9 A~ Cr'8m ,slnlll'il Df! ,Pl'\e'~fe'flll memJitJ'no· 9!5ifiJ'ca ~:w::"iI21iiu:t. Smull, jlloomr.l:'[lIh1r, grR[i:l~:iC~~tl~ve'('OO. cdlQdll~ ami b~:lt~]H can hI:! o:b.sI!!J'!i<'I!iI ln ~hl;s S~t£rut! £I. iChruracte.rimic "ppcarflJilCC (If P/,fIj'l:flfJJtr me&mul!Jgtllir:li. a~ .P.Ailiw;mtul1a me,p:B]"fnli:!g.e.iitk~ oml ,1I'~~1ia IbI'o'!CI<d D!$)li"io Tb~ smt~1. dark.~Rri_I;!i~ eelerues •. OO'sSsta:Llt ~c Inlll~ IlQ~'C111 (1~e&S thilllt UJi :LlllM zone or lllhibitkll~' mld""'d[l~mll$il1" 1Ml] '~!JOl'iesce'OO1C1k'fied untie[' ulird.vcoiE!l ligJllt{1N~ l;;)J:.m1l), C," ~ ~ntiii, mela"lo'!'l1q'Uin~'c:a onl 8~~ ~!I!,\!OO '''''B!lU', Aftersevtll'rulnru'C da,lI'S oriuclLlbI:iHo~~. these o(l~on'f,es ,",'III ta:l.ie Oil! @.dark'brou,'11, to I:ili:l,c:~!I; oolut. diL!llc [lJc.;;l!'i~imil.ttl(lJll ol'hftillo m ~he medlum.

1 0"'10B\a~lfii.nJl'd'Ii!:Iii!fI'ag'llJll! ,Omld ,P~~~Q"~! Ji:!'1!iI,a-nl.Ji!'!Il!'svnl'aa' 'DITIJ Baa~~~!!i!l~!f!1 !f!"'~WI.n t,DBE.t!ag.a;r:" 1,fuic ,Pm<'Or~!li1 spseies W<1l'S slJieak~tl on lilifJI I.:Ip~c· h.llr Q.IT tim plate, but grjJwl·h\v~!llnhlblled. The i&lderoI!It·;,: /lqili~ :~ro,"jS pmj!.l;s~l)·; d!ll~IOI!j'lrtg a ~1~{':[] I :I,l iol~' ilf.eeiipl~l.Il'i3 tr.i lfile agar surroundmg the colonjes,

.' .,
r/ ,.
'. ... '.'
. "
' .
• B

113·''111 ,~, :At.OJiil!!:;WF um rluclL-ah.lm 'o:n SAlC'8.U blvQ;G ,~ggr'. The dry. \!I'nile. b!'rp.:.:;tdCJ;'~mbhkc 'nl!on!e-_, tone c 10 C" 'M.Bllt of th] species) and the ::1.1] ~ul:Jiotl c dl!ik p::lJLtern (bLIII.I:UIIycio· and constlJJ::j,-S\1~(lt.ible; ValnCQllll'Cttl,~res.L.st.ohilt) ;lim su~ges[J.w of E m,!CJt'lirriml, If _[[Iclol.Breagerrt 1:\1 l'1·cl!ticcl to t~Jjrr exl:n.li_Uer p.ap,.t cllbJ ... onthe lPll..-te. the dISk \'IIiUl. Urn bl I bee.me: :1111'5 sp.er"il:lS is iln~ole1Xls1thrc. iii" Oil'. m si.~il!l ''!iJf Fll'sabBiDilU"IU:m nl!O'!m" ftlm 4)(1.I50J. p<JJI stl-ljr]UI'ilj;l. IOU1g. Jenner. gram-i1es~tiwlbl:lcilli with lapL;.'t'cillpointeri ClUJ. demun:s1:r.alli:l:Jg the charactcrlstle illPJ!.!r.!fIIT,f;fli.et" ul' F'JJS'oh!Tflerl!'m~ ~~JtI.r..d~f;ml.

"0 .. 12 P.ptiai·~ .. ptuc;:~.. anaucn::libl'.us ,~" Sr,uo.lla bmDOcill ,a9ilr. This gram-uegsttve cr CU dlsplay IIl€rl,illlml.siz:ed ilfOiS C010Lli£s. ror>~:l:mfiCli! D.Q kmul myeln and oolh.tLI'I, ;]11 d gjlstl~~~~illt,y to ltilUl!l..:umyc·i n,

'10"''',3 P4!Jpfa;a;iI'F'&pNC'Oce;uwo maga .. .::, ,g~1 Sru.,C'elhi1llblaad 8g)illlil!". "1'h18 pecic$. bas.liny colontes but CralJII staln yidti.:> large, m:bcrOC{lCCtiS-ilP~;:JJr.i.lI:tig. iil(HII11-posHh-e oocct, Lt i~ suscepiilhk to tani;ln'L}'!.:in and '\'~mtl,Xlrnj!cln and resl 'la[l( ~omli:stin.

1liEll.114 SQ!i:I!IIIm pal¥8n."tl1u_ -ulfonat. 'E!l:P~I) r'Q" I!l!Jsct~nc8' at! P!IQIMi!I~kic:~cu\lll!' m_ P~fii". llJjke Po WWfI'i oi~,:s. P. , tlig.m~:;;: Is rest tant to Sf'S ..

lI'l;)-l.!!5 Cg~g!liIjiil;!!ii!' 011' ~mlI~:Jt!-ii!!ef.ri(!!J7!1 ,aenss .g!~ 6!!\""~",,,tl!!;l! Ib'!.g'g!:!i I1l!D.f!!f". -rhC:SCyt!iling ctllh.JiJtie:; ii1n" Slnlall,.wd w.hll.e to gr-dJ"-\lI'!IH~; h!lYI!iU'i't.r.. <LS lhtzy' ~g~. [~II!Y can i~PP(:U }I~"llmrVL'll h, 1"J:m~ haw im~ml ruJe~ red lUi as mfl,sil:!wi;lir: mph tlleil."oidt; [,~[;;JiI]~, l\i'herJst;li[lI!1~cl. ~ml examined mlcrosccptnllly, ItJUlY resemble diphthe[l]iju;s. PfD~rilJ/l!bt[t-1 frirililil frn!(!,~' are UI' dol-posnlve, m; seen on lhis plate, and C!mI13SB-'P,OO!ti'l;\~. Whim an ,Iilimu:::ro[)fc diphtheroid is cata I use- u rndilld.nl.-,ll!rl.~iH~'~', il can be 1]£e:>U:L1I~:rl~vely ld~Il,LWi'ed as fRrp.rjrn'fjoor,l!r~·hrrr'! (Ifmrn,

iII,G-1,g, RlII'!!,\liS:AIIQ' CAMP tQut to .. · JI'!'I~!l!mpilllll!l'" IdenWi~atIii~Uf! ,~", ~MdI~ peri!l"m,O'li'ff.Ii!I' C, [Jl'r" fi',i'~WI1~ ~ iltrc<l~()~·!i:'tlli.c:,~UY,il!.Il(] S"i:;rQM(l('.o~'cjlg ,(lYfiimJilft' Ib ,~"ltl.'i.;,j).kk."{l hudi'imuaJl~i in llti~ lcsl, '('11 Ii' hCimolY1>i:> of t~c dtl!!Ot ddl un!1 ls ~)'iu~rl!lsl k.lI]y en hancad by I,he' b~~'I1Il]yslli or lhe ::;Lr[;!l1tlji~:!jJI!."c!!I;S in 1m ;![ntil'lrvbeij[d·5IL~,~r:II -. pattern,

110 .. 1 a ar,am filidaln at' e~'aatil'lidJ'U'.m iII:PQ!i:'i!l;!ll;l; ,( x1:2H.5iO'~. C~1!I.\SlJre P,ii:l[aU~l • .sidl:ld r lung. thirr., ~rrrm·\m riahk', ,<I!.:rd OOm,e show ~III'UU~Ll ends imiit"aiiiJ..l:: of. spo~ [:orlJurllOi~,

110-17' G'J'@~~'~m' p~~I'I\S1ei!n~ an 6'ftl'iCG11B b'Joudl sgar:. CoL'arl tss db;~I.tlJr the douo.~~-Wi'l~ uf beta hemOZYSiIf; t~i!!ll('~ri or Ihis i'i(JOc1~:5. Colonles ~re l~rge ~\'iil~ FlikEd (;,rH'ilt'lJrS .m;[~ ifoog:-tIlrlr ~dl!iill .e rt~r 4fl h mil rs.irl(!IJiDation.

1 g·lIIBC'lslrll"ld1.1M1 F.l'e~.IlI'NI' o:n egg'Y'aUc; ,i!I![!illlr.. Cohmlm; and S'LII[[(JuudiIl1]. medium ui.~lJ)l,,}y 1.1] e ec'i;:pm:u1iiTlJg W.hLI,e IJrer:ipit;llm' sl:ilrrrrlD~0l4~. by 1(lC'H:bi[] ase producHum .• InriUi!l or e. !i'i!'rft'.rllfj\('Jt~.

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