e:> Pergamon

~
Wat. Sci. T«It. Vol. 35. No.6. pp. 91-99.1997.
1997 IAWQ. Pubhshed by Elsevier Scicli« Lid
Pnnted in Oreat Bnlain
0273-1223197 SINlO + 0-00
PII: 50273-1223(97)00099- J

ORGANIC CARBON AND NITROGEN

REMOVAL IN MOVING-BED BIOFILM
REACTORS
G. PastorelIi*, G. Andreottola**, R. Canziani*,
C. Darriulat*, E. de Fraja Frangipane* and A. Rozzi*
• D.I./AR. - Sez. A!nhientale. Politecnico di Milano. P.ua Leonardo da Vinci 32,
1-20133 Milano. Ilaly
•• Dipartimento di Ingegneria Civile e Ambienzale. Universita deg/i Studi di Trento,
Via MesUlllo 77. 1·38050 Trento. Italy

ABSTRACT

Pilot moving-bed biofilm reactors (MBBRs). fed on primary senled wastewater. were used in order to study
organic carbon removal and nitrification. Nitrate uptake rate (NUR) tests were performed by feeding sodium
acetate and potassium nitrate to • bench-scale moving-bed balCh biofilm reactor. In both experiments the
same polyethylene biofilm carriers were used. Both particulate and filtered COD removal rates appear 10 be
proportional to the corresponding loading rates. Particulate COD removal is the net effect of adsorption onto
and release from the biofilm surface. Filtered COD removal is the sum of the influent filtered COD removal
and the removal of hydrolysed colloidal COD. Filtered COD removal rates could not be evaluated with a
kinetic expression because back-diffusion from biofilm is not always negligible. Nitrification tests.
performed at oxygen limiting conditions. show that the reaction rate was nearly first order with respect to
dissolved oxygen due to liquid film diffusion. Denitrification balCh tests showed denitrifICation rates very
close to other reported data. Since the process proved reliable and easy-to-operate. it is suitable for
application to small WWTPs. either in designing new plants or in upgrading existing overloaded activated
sludge systems. @ 1997 IAWQ. Published by Elsevier Science Ltd

KEYWORDS

Acetate; denitrification; hydrolysis; liquid film diffusion; moving-bed biofilm reactor (MBBR); nitrate
uptake rate (NUR), nitrification; organic carbon removal.

INTRODUCTION

Biological processes based upon suspended biomass (Le., activated sludge processes) are effective for
organic carbon and nutrient removal in small and large wastewater treatment plants. But there are some
problems of sludge settleability and the need of large reactors and settling tanks and biomass recycling.
Especially the fIrst item could give rise to serious operating problems in small wastewater treatment plants,
due to lack of operation.

Biofilm processes have proved to be reliable for organic carbon and nitrogen removal without some of the
problems of activated sludge processes. Moving-bed biotilm reactors (MBBRs) were chosen because,
compared to the available fixed-bed biofilm reactors (biofilters), they are characterized by low head losses,
91
92 G. PASTORELLI ~, aI.

no filter bed channeling, no need of periodic backwashing and can be used to upgrade existing overloaded
activated sludge plants without the building of new tanks.

Low-density polyethylene (density slightly less than 1.0 g cm- 3) biofilm carriers were used. They consist in
small cylindrical elements (diameter 10 mm; height 8 mm) with an internal cross and small longitudinal fins
on the outside surface. The filling ratio (volumetric flliing in empty reactor) can be increased up to 70%,
corresponding to a theoretical specific surface area of 500 m 2 m- 3. Since the biofilm growth mainly occurs
in the protected internal faces of the biofilm carriers, it was assumed in the calculation an actual specific
surface area of about 350 m2 m- 3.

These new Norwegian patented MBBRs have been already successfully used in the treatment of dairy
wastewater (Rusten et ai., 1992), NSSC corrugating paper mill wastewater (Broch-Due et ai., 1994), potato
chips wastewater (0degaard et ai., 1994) and municipal wastewater, particularly, in small wastewater
treatment plants (0degaard et ai., 1993) and for nitrification (0degaard and Rusten, 1993; Hem et ai., 1994;
Rusten et ai., 1994, I995b) and denitrification (0degaard and Rusten, 1993; Rusten et ai., 1994, 1995a).

MATERIALS AND METHODS

Ex,perimental Proeramme

Organic carbon removal and nitrification were studied in pilot aerobic MBBRs, while denitrification rates
were evaluated by means of nitrate uptake rate (NUR) tests performed in a bench-scale moving-bed batch
biofilm reactor.

The pilot-plant, fed on primary settled wastewater, was operated for 88 days with two aerobic reactors (OX
and NITI) in series: OX reactor achieved organic carbon removal and NITl reactor achieved residual
organic carbon removal and, mainly, nitrification. The plant was operated for the following 43 days (from
day 89 to day 131) with three aerobic reactors (OX, NITI and N1T2) in series, because temperature
decreased and it was impossible to maintain a stable nitrification in NITl reactor only.

The bench-scale reactor, fed on sodium acetate and potassium nitrate, was used to perform seven NUR tests
using a small number of biofilm carriers taken from OX reactor, when it was converted to completely mixed
pre-denitrification mode, after day 131.

Description of the Ex,perimental EQuipment

Pilot-piant. A simplified flow-sheet of the pilot-plant is shown in Fig. I. Technical and operating data of the
pilot-plant are reported in Table I. Further details can be found in Pastorelli (1995).

A constant flow rate of 30 I h- I (0.720 m 3 d-I) of primary settled wastewater was fed to the pilot-plant
through a first completely mixed not aerated tank (IN) where hourly fluctuations of the concentration of
each contaminant were equalized (HRT=13.33 h). Therefore, it is possible to assume that the wastewater fed
to the MBBRs had daily fluctuations only.

The three MBBRs were completely mixed aerated tanks. Air flow rates can be adjusted separately for each
reactor.

No settling tank was provided, because one aim of the research was to study the behaviour of each MBBR
by means of mass balances of each reactor, taking into account also suspended solids and particulate COD.
Settleable solid tests in Imhoff cone were performed on the effluent of each MBBR.

The pilot-plant was installed outdoors, inside a metallic pre-fabricated box without thermal insulation.
 Organic caJbon and nitrogen removal in moving-bed biofilm rcaclon 93

Bench-scale reactor. A simplified scheme of the NUR test perfonned on the bench-scale reactor is shown in
Fig. 2.

The bench-scale reactor was a 2-1 beaker (bioreactor volume 1.8 I), completely mixed by means of a
magnetic stirrer. Temperature was controlled during each test. In order to obtain a good mixing of biofilm
carriers, a low filling ratio was chosen, corresponding to an actual specific surface area of 50 m 2 m- 3. Total
biofilin surface was 0.088 m 2. Biofilm carriers were taken directly from OX reactor, because of very high
percentage of heterotrophs, immediately before each NUR test.

IN ox NIT! NIT2

q I q
II o\it .. $
III
tP,,~ .. ...
q
ijo)"',...,. ...
q olllu..,'
wasklwatar
I ilJ

;0 +Io~ 'to:.,
c::b
~o ..... 900."0$
;
.......
~o ~o ~ ?o'if

• '!
.<11
(I . . 0

:'
"

.<11
$ t" +0""
.'" t .. .... :t .0'"..
... t° ...

I
air~
I I
Figure I. Flow-sheet of the pilot-plant

NUR
oodUIlII t-o , t-o pulaAsium
ICClIIOJ i r;-"~'IO
*' ttl' of>'"
o+t ,tfI "'"
0, _
.........
... ~ ~- 0+-
... ... .
Figure 2. Scheme of NUR lest

Table 1. Technical and operating data of the pilot-plant

Technical and operatmg data

IN OX NITI NIT2
tank reactor reactor reactor

Height [m] 1.00 0.91 0.62 0.91

Diameter [m] 0.75 0.57 0.51 0.57
Bioreaetor volume [ml ] 0.400 0.090 0.060 0.180
Filling ratio [-] 0.47 0.40 0.16
Actual specific surface area [m2 mol] 233 200 78
Biofilm surface [m2] 21 12 14
Influent and effluent flow rates [ml d-I] 0.720 0.720 0.720 0.720
Hydraulic retention time [h) 13.33 3.00 200 6.00
Mechanical mixing yes no no no
Aeration no yes yes yes
Operating period [d] I to 131 Ito 131 I to 131 89 to 131
94 G. PASTORElLI etal.

Wastewater Characteristics

Pilot-plant. The pilot-plant was located at the Varedo wastewater treatment plant (near Milan), which is a
WWTP (150,000 PE, 110,000 domestic and 40,000 industrial PE) fed on a combined sewer system.
Characteristics of the primary settled wastewater fed to the pilot-plant are reported in Table 2.

As Table 2 clearly shows, there were large fluctuations of concentration of the contaminants, especially
those related to the suspended fraction. Fluctuations of concentrations were due to the periodic change from
dry to wet weather conditions and vice versa, but fluctuations of concentrations related to suspended matter
were also due to the occasional overloading of the primary settling tank.

Bench-scale reactor. The bench-scale reactor was batchwise fed on a suitable solution of sodium acetate
(8(}-120 mgCOD I-I), potassium nitrate (15-25 mgNOf-N I-I) and micro- and macro-nutrients needed for
bacterial growth. Initial CODIN ratio was set to values around 5 mgCOD (mgNOf-N)·I.

Monitorin~. Samplin~ and Analyses

Pilot-plant. Temperature and dissolved oxygen were measured in each MBBR every workday, immediately
before sampling. Air flow rates were monitored using flow meters.

Grab samples of influent wastewater and effluents from each MBBR were taken 5 days a week in the fIrst 45
days and 3 days a week in the following period. Since the hourly fluctuations of the concentration of each
contaminant were equalized in IN tank and MBBRs were completely mixed reactors, it was assumed that
grab samples were not much different from daily average ones. Nitrogen mass balances through the pilot•
plant clearly showed that the approach was correct.

The samples were analyzed immediately after sampling for the parameters shown in Table 2.

Bench-scale reactor. Temperature and dissolved oxygen were continuously measured during NUR tests.

Grab samples were taken from the batch reactor every 30 minutes. Grab samples of influent synthetic
wastewater were also taken directly from the reactor immediately after feeding.

Table 2. Characteristics of the primary settled wastewater fed to the pilot-plant

Parameter N Min. Max. Avg. St.dev. Coeff.var.

TSS [mg 1.1] 55 16 1132 176 224 1.27

Settleable solids [mll· I ] 48 0.0 46.0 3.6 8.4 2.32
Total COD [mgl· I ] 49 51 1113 276 171 0.62
Filtered COD [mgl- I] 40 30 281 142 55 0.39
Particulate COD [mg 1. 1] 38 16 1038 155 183 1 18
Total N [mg I-I] 13 18.9 50.4 34.0 8.0 0.23
TKN [mg I-I] 17 10.8 50.1 32.3 94 0.29
NH.t+.N [mgl- I ] 54 4.8 47.2 26.5 7.9 0.30
NOf-N [mgl- I ] 29 0.1 1.6 0.3 0.3 0.85

Analytical methods. Total suspended solids (TSS), settleable solids, COD, ammonia-nitrogen (NH 4+·N) and
TKN were measured according to Standard Methods (APHA, AWWA, WEF, 1992), and nitrate-nitrogen
(NOf-N) according to Italian Standard Methods (IRSA-CNR, 1979). Whatman GF/C fl1ters were used for
filtratinn nf .,.mnlPQ
 Organic carbon and nitrogen removal in moving-bed biofilm reaclon 95

Particulate COD (partCOD) was calculate as difference between total COD (totCOD) and mtered COD
(fIltCOD), while total nitrogen as sum of TKN and nitrate-nitrogen.

RESULTS AND DISCUSSION

The operating conditions and loading rates of each MBBR are reported in Table 3.

The experimental programme aimed to study the kinetic behaviour of the reactors for organic carbon
removal and nitrification. Therefore the reactors were not always operated to achieve the optimum removal
efficiencies, but were operated in order to evaluate the maximum removal rates in different conditions. For
example, nitrification was studied at different dissolved oxygen concentration, different organic carbon
loading rates and excess ammonia.

The wide range of temperature obtained was due to the absence of thermal insulation. Therefore, it was
possible to evaluate the pilot-plant behaviour in different climatic conditions over a few months.

O[~anic Carbon Removal

Total, particulate and filtered COD removal rates in both reactors vs the corresponding loading rates are
shown in Figs 3. 4 and 5. These performances do not take into account any settling which could improve
total and particulate COD removal rates.

Removal rates appear to be proportional to the corresponding loading rates but this is the result of two
different removal mechanisms. Particulate COD removal is the net effect of adsorption onto and release
from the biofilm surface. These actions are purely physical, since the observed removal is not dependent on
temperature.

On the other hand, filtered COD removal is clearly a biological process, since the highest removal rates have
been observed at the highest temperature. However, filtered COD removal is the sum of the influent filtered
COD removal and the removal of influent colloidal COD that has been hydrolysed in the reactor.

Table 3. Operating conditions and loading rates of each MBBR

OX reactor NIT! reactor NITZ reactor

Parameter
Range x:tSD (n) Range x±SD (n) Range x±SD (n)

T [0C) 7.I+Z4.5 16.0%4.7 (54) 7.1+24.5 16.2±5 0 (54) 6.2.,.11.5 9.J±1.S (14)
DO [mg02 I-I] 0.5+6.9 3 9±1.5 (53) 2.9+10.5 5.9±1.9 (54) 95+12.5 Il.I±O.7 (14)
Loading rates:
TSS [gTSS m-2 d-I] 0.5+35.0 59±7.1 (55) 1.3+42.3 8.9±8 8 (48) 4.3+24.1 9.3±6.1 (9)
TotCOD [gCOD m-2 d-I] 1.7+38.2 9.8±5.8 (49) 2.9+44.1 13.3±IO.O(44) 65+30.3 13.4±7.9 (7)
FiltCOD [gCOD m-2 d-I] 1.0+9.6 5. I±Z.l (40) 0.4+13.3 4.2±2.3 (36) 30+7.5 4.6±1.8 (8)
PartCOD [gCOD m-2 d-I] 0.5+35.6 5.5±6.2 (38) 1.0+41.6 IO.2±1O.2 (36) 3 5+25.9 8 8±7.7 (7)
NH4+-N [sNH4+.Nm-2d-l] 0.0-2.5 1.4±0.5 (46) 0,9+1.9 1.5±O.J (II)

Research on kinetics of hydrolysis of non-diffusible COD in biofilms is still at its early stages and a fully
developed kinetic expression is not yet available. Moreover. it is still unknown whether non-diffusible COD
is mainly hydrolysed in the bulk liquid or on the biofilm surface (Harremoes and G/jnen~. 1983; Rohold and
HarremoCs, 1993; Larsen and Harremoi!s. 199480 b; Ro. 1995). Finally, daily fluctuations of COD
concentration in the feed were so high that effluent concentrations may have varied according to different
96 G. PASTORELLI el al.

causes, other than removal. such as back-diffusion from biofilin during low loading conditions following
higher loads.

Although this is clearly a purely empirical approach, still it may give valuable predictions on the
performance of this type of biofilm reactor.

In Fig. 5 (OX reactor), two points lay far away from the others, but since they correspond to high flItered
COD loading rates and low dissolved oxygen concentration (<2 mg02 1-\). they have been excluded from
the linear regression. This may indicate that the linear link between loading and removal rates holds when
dissolved oxygen in MEBRs is more than 2 mg021-\ and flItered COD loading rates are less than 8 gCOD
m-2 d-'.

Filtered COD removal rates were corrected for temperature with a simplified van't Hoff-Arrhenius
coefficient of 1.043. This coefficient gave the best linear regression (with the least-squares method). Since
the range of temperature spanned from 7.1 to 24's"C. this may explain why this value is slightly higher than
other data reported in the literature for organic carbon removal.

Nitrification

Nitrification tests were performed at oxygen limiting conditions, that is at high ammonia concentration (>4
mgNH 4+-N 1-\). The dependence of nitrification rates on dissolved oxygen concentration is shown in Fig. 7,
where also the influence of the filtered COD loading rates is reported, due to their effects on competition for
oxygen between heterotrophs and autotrophs.

Figure 7 shows that the reaction rate was nearly first order rather than half order with respect to dissolved
oxygen. This may imply (Harremoes, 1978) that liquid film diffusion is the limiting step of the process,
according to other reported results on the same biofllm carriers (Hem et al., 1994).

Figure 7 also shows that there is no nitrification when dissolved oxygen concentration is less than 2.08
mg02l'l. This is probably due to the higher heterotrophic activity in the outer layers ofbiofllm (Wanner and
Gujer, 1985) and agrees well with data reported by 0degaard and Rusten (1993) and Rusten et al. (1994).

Nitrification rates were corrected for temperature with a simplified van't Hoff-Arrhenius coefficient of
1.124. This coefficient gave the best fit correlating nitrification rates to dissolved oxygen at low flItered
COD loading rates (<3.5 gCOD m-2 dol). Again. the wide temperature range (6.2-24's"C) may account for
the high value of van't Hoff-Arrhenius coefficient. if compared with other data reported in the literature for
the same biofilin carriers (1.107. Hem el al., 1994; 1.090 between 7 and n'e, Rusten et aL. I995b).

Although a linear link between nitrification rates and dissolved oxygen can mean high energetic
consumption, still dissolved oxygen concentration may be used to control the nitrification process at high
ammonia loading rate or at low temperature.

Denitrification

NUR tests with excess acetate (CODIN-5) were performed and results are shown in Table 4. Very little
acclimation was required for denitrification to start (0.5-1.0 h), even if biomass came from six-month
aerobic operation in the pilot-scale reactors. This was easily explained by observing that some denitrifying
activity took place even in the aerobic reactors, probably in the deeper parts of the biofilm (simultaneous
denitrification: see also Riemer and Harremoes, 1978 and Chen et al., 1989). In fact nitrogen mass balances
in OX reactor showed that 3.3 N03--N 1-\ were produced, but only 0.14 NOf-N 1-\ were present in the
effluent Some denitrification was present in NITl reactor also. but only 0.9 mgNOf-N 1-\ were removed
(from 4.9 to 4.0 mgN03'-N 1-\) due to the scarcity of available organic carbon.
 Organic carbon and nitrogen removal in moving-be d biofilm reactors 97

8 81 NIT2
:! ~ 6
NITI

.. . . .. ' .. I~ 41 ..
.' ~C2 ••• : .... :
t-I- -.........;-l.~'r-.---_ . . . . .- -•
8o ~ -2 t,
08 0
2 4 6 8 10 12 14 16 18 20

- -4'

totCOD Ioadmg rate [gCOD/m2ld) totCOD loadmg rate [gCOD/m2ldj

pilot-plant
Fig. 3. Total COD removal rate vs total COD loading rate in each MBBR ofthe

3
i OX

.-
;;~2

i~ 1
0 0

i~-1
0
1
, 2 3 4 6 7 8 9 10

·2
partCOD loading rate [gCOD/m2ldj partCOD loading rate [gCOD/m2ld]

MBBR ofthe pilot-plant

Fig. 4. Particulate COD removal rate vs particulate COD loading rate in each

i~,
.... ""S 4
~o
80 3
E~ 2
OX :
: I'
, '
00<2.,.0211"1
2<1)(}<4 ,..0211
-iM~~
4
"0 3
~8 2
'I NITI
NIT2

.
.~:. •
I: ~:::~i
r .. , -DO--9..,o21t I

:::.:;n;
o~ I
1-_61l111
• DO -9,..0211

@~ ~ . . . . .- - - - - - - Ii :
O~01-7""~i-----­
(,)0
~ ~ -I
7 8 "-,- - - - - '
is ti 0 I 2 3 4 , 6 7 8 910 -_611 i,.

filtCOD loading rate mtCoD loading rate

[gCOD/m2ld] [gCOD/m2ld)

ofthe pilot-plant. Regression

Fig. S. Filtered COD removal rate vs filtered COD loading rate in each MBBR
reactor (limited to DO>2 mg02 /-1): rtiltCOO.2o "C=-O. S I +0. 70·fiItCO D LR; k-r= I.043; r2a0.83;
analysis on OX
reactors: rtiltCOD, 20'c-o.92 +O.S9·fiI tCOD LR; kr=I.043 ;
nz24. Regression analysis on NIT I and NIT2 2
r -o.65; nz 33

1~;'I
B:O
OX: .:
!:==ml ~~~i :f "0
0 0 4,
NITI
NIT2.
• , .
i • lJOo.4,..o211
;. UX)..6 11ll1U2iIj
~~ 2: •• : • ",[)0<..6q02 !t I 2
II.. ' ., !
I ..
.
• 6·DO'9,..o~:
,
~ 1;
o~
<:> ••
! " oo:-ti IIW02t1 Q r,
o~
..... ~ -.'
0" I . 00-9,..0211 1
~~
90 120 UO
"

1- - - - - - - •
0 ... ""
..... -2 '30 60
o 30 60 90 120 UO
effiuent filtCOD [mgCODllj effiuent filtCOD (maCODllj

the pilot-plant
Fig. 6. Filtered COD removal rate vs effluent filtered COD in each MBBR of
98 O. PASTORELLI etat.

'iii t;;' 2.S

~ ~ 2.0 i
i
• &lIL"ODLIl3S
...."lDmlId :
8~ZI.S
'il ~ 1.0
~
'a
i.
t:5 O.S
::'§ 0.0 l--~_"""":"
,. ~:::I
o 2 4 6
I
_

8 1012 14 i
bCOD1JL.>60
1"00'",'"
I

dissolved oxygen 1 -..........

[mg02ll)

Figure 7. Nitrification rate vs dissolved oxygen concentration in NITI and NIT2 reactors. Regression analysis
(limited to filtCOD loading rares<3.S gCOD m-2 d-I): r nit.20·c=O.30(DO-2.08)O.94; k~1.124; r2:0.79; n=6.

NUR tests gave useful indications on the maximum achievable removal rate with a rapidly biodegradable
carbon source. The results are comparable with data reported by Rusten et al. (1995a) with the same biofilm
carriers and also with data obtained in a continuous sand filter (Bonomo et al.• 1995).

Table 4. Results of NUR tests with acetate

Test T Nitrate range Denitrification

[0C] [mgN03'-N I-I] rate
[gNOf-N m-2 d-I]
I 20.3 14.0 - 6 0 2.1
2 18.5 18.3 - 5.3 1.7
3 18.9 16.0 - 6.1 1.8
4 18.4 19.1-6.6 2.5
5 22.8 21.3 - 7.2 2.2
6 20.6 23.6 - 7.9 2.4
7 21.1 20.9 - 7 1 2.9
Avg. 20.1 19.0 - 6.6 2.2
St.dev. 1.6 3.0 - 0.8 0.4
CoetIVar. 0.08 0.16-0.12 0.19

CONCLUSIONS

The process was studied in view of its application to full scale WWTPs. Since they proved reliable and easy•
to-operate (no clogging problems, few suspended solids in the reactors and easy to settle or mter out, simple
and rapid start-up. etc.). it is suitable for application to small WWTPs. either in designing new plants or in
upgrading existing overloaded activated sludge systems. In this case they only need some adjustment of the
aeration/mixing devices and the introduction of a suitable sieve at the outlet of the reactors.

MBBRs may be used efficiently to remove organic carbon (in part, also the colloidal fraction) and nitrogen.
The nitrification process may be simply controlled by means of dissolved oxygen concentration.

ACKNOWLEDGMENTS

The authors wish to thank Mr. Z. Giannoni and Dott. A. Losi (Consorzio Provinciale di Depurazione delle
Acque del Nord Milano, Milan, Italy) for the kind cooperation at Varedo wastewater treatment plant, Prof.
H...~degaard (Norges Tekniske H~gskole, Trondheim, Norway) and Mr. O.H. KoIkinn (Kaldnes
MilIJf/lteknologi AS. Tmsberg. Norway) for supplying the KMT-biofilm carriers, Mr. E. Breda (GMC S.p.A..
 Organic carbon and nitrogen removal in moving-bed biofilm reactors 99

Milan. Italy) for supplying the aeration system and Dott. Ing. G. Favali (Smogless, Milan, Italy) for
supplying metallic air pipes and plastic fittings. The authors wish also thank the staff of D.I.I.A.R. - Sez.
Ambientale (Politecnico di Milano, Milan, Italy) and. namely, Mr. F. De Pascalis, Dott. P. Giavazzi, Mr. G.
Gurrieri, Ms. L. lollo, Dott. L. Nannariello and Dott. A. Perosa for helping in the operation on the pilot•
plant.

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