ARTICLE IN PRESS

WAT E R R E S E A R C H 42 (2008) 1083 – 1092

Available at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/watres

COD fractionation of tannery wastewaters—Particle size

distribution, biodegradability and modeling

Ö. Karahana,, S. Dogruela, E. Dulekgurgena, D. Orhonb

a
Environmental Engineering Department, Faculty of Civil Engineering, Istanbul Technical University, TR-34469 Maslak, Istanbul, Turkey
b
- ankaya, Ankara, Turkey
Turkish Academy of Sciences, Piyade Sokak No. 27, 06550, C

art i cle info ab st rac t

Article history: This study aims to establish the scientific link between particle size distribution (PSD) and
Received 18 December 2006 biodegradability of different COD fractions of tannery wastewater, by means of sequential
Received in revised form filtration/ultrafiltration, respirometric analysis and model evaluation. PSD profiles were
24 September 2007 determined in physical segregation experiments, using eight membrane discs, each with
Accepted 1 October 2007 different pore sizes between 2 and 1600 nm. Biodegradability-related COD fractionation was
Available online 6 October 2007 determined at each size interval by model simulation and calibration of the corresponding
oxygen uptake rate (OUR) profiles. Activated Sludge Model No. 3 (ASM3), modified for direct
Keywords:
growth on hydrolysis products, was adopted for evaluation. PSD analyses defined a COD
ASM3
fingerprint with two significant portions at the two ends of size distribution, with 60% of
COD fractions
the total COD at the particulate range, 25% at the soluble range and the remaining 15% well
Modeling
distributed among the colloidal range. Comparative evaluation of the sequence of OUR
Oxygen uptake rate
profiles yielded values of applicable model coefficients. It also enabled the assessment of
Size distribution
size distribution for each major COD fraction, as an original tool for better interpretation of
Tannery wastewater
specific biodegradation characteristics of the selected tannery wastewater. Results also
revealed a very slowly biodegradable/residual particulate COD component with a
significant inhibitory effect. Model-based evaluation of the OUR profiles enabled quantify-
ing the impact of inhibition in terms of changes in rate coefficients for growth, hydrolysis of
soluble COD and endogenous decay.
& 2007 Elsevier Ltd. All rights reserved.

1. Introduction of effective removal requires elaborate evaluation of biode-

Tannery effluent is a strong wastewater with complex
characteristics. It is associated with a high level of organic
pollutants commonly characterized with chemical oxygen
demand (COD) concentrations above 3000 mg L1, together
with significant amounts of inorganic compounds such as
trivalent chromium and sulfide, capable of exerting toxic or
inhibitory effects on biological treatment (Kabdasli et al.,
1993; Orhon et al., 1999a; Carucci et al., 1999). Stringent
effluent limitations inevitably prescribe biological treatment
for this level of organic content. Consequently, achievement
gradation characteristics and possible inhibitory effects
imposed on the degradation process due to the nature of
tannery wastewaters.
COD is a useful collective (lump) design parameter reflect-
ing the total organic content (CT) of a wastewater. Yet,
it has inherent deficiencies when used alone as it covers
not only biodegradable organics but also biologically resistant
and refractory compounds and provides no information
on significant organic fractions with different biodegradation
kinetics. This missing information is apparently vital for
the tannery wastewater which embodies a wide spectrum

Corresponding author. Tel.: +90 212 285 65 40; fax: +90 212 286 7913.
E-mail address: okarahan@ins.itu.edu.tr (O. Karahan).
0043-1354/$ - see front matter & 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.watres.2007.10.001
 ARTICLE IN PRESS
1084 WAT E R R E S E A R C H
of various organics. Previous studies have reported
that tannery wastewaters include total biodegradable
COD portions (CS) comprising different fractions with
markedly distinct biodegradation rates and inert COD
fractions play an important role on the extent of treat-
ment performance (Ates et al., 1997; Orhon et al., 1998,
1999b).
The recognition of different soluble and particulate
COD fractions not only provided a major breakthrough in
the conceptual understanding and modeling of activated
sludge (Henze et al., 1987) but also introduced respirometry
as the main instrument for assessing COD fractions as an
integral part of wastewater characterization and modeling
(Spanjers and Vanrolleghem, 1995). While the interpreta-
tion of the oxygen uptake rate (OUR) profiles enabled
distinction of readily and slowly biodegradable COD fractions
(Dold et al., 1980; Ekama et al., 1986), and assessment of
major model coefficients (Kappeler and Gujer, 1992; Avcioglu
et al., 1998), COD fractionation has been primarily based
on a single size, that is, the routine filtration size (1600
or 450 nm) used for differentiation of particulate and
soluble COD components. Introduction of COD fractions with
rapid and slow hydrolysis rates, although giving a clearer
picture of the biodegradation profile within the soluble
(filterable) range (Henze, 1992; Orhon et al., 1999c), did not
include any additional information about particle sizes
involved.
Research effort has recently been directed towards particle
size information for a better understanding of COD fractiona-
tion and related biodegradation patterns (Sophonsiri and
Morgenroth, 2004; Dogruel et al., 2006; Dulekgurgen et al.,
2006). Combining physical particle size distribution of COD by
means of sequential filtration/ultrafiltration with parallel
OUR experiments and model evaluation is the next step for
a better interpretation of COD fractions and biodegradability.
Integration of these two approaches would also provide
useful information for optimization of available treatment
techniques indicating which size fraction is more important
within the overall organic content and how the biodegrada-
tion characteristics of COD fractions differ in terms of
treatment requirements.
In this framework, the objective of the study was to
determine the size distribution profiles of major COD frac-
tions in tannery wastewater. This goal was accomplished
by means of sequential filtration/ultrafiltration, together
with parallel respirometry and modeling for exploring the
relationship between physical segregation and biodegradabil-
ity. Accordingly, the raw wastewater and the permeates
from each sequential filtration/ultrafiltration step were sub-
jected to respirometric tests and the resulting OUR
profiles were evaluated by model simulations to assess the
respective COD fractions and biodegradation characteristics.
Tannery wastewater was selected mainly because it has
been well studied and intensely utilized for testing and
exploring new mechanisms such as dual hydrolysis
phenomenon (Orhon et al., 1999b). In this study, the
complexity of the tannery wastewater also required modifica-
tion of the basic structure of Activated Sludge Model No. 3
(ASM3) for better interpretation and calibration of the OUR
profiles.
42 (2008) 1083– 1092
2. Materials and methods
2.1. Study area
The tannery wastewater investigated in this study was taken
from Corlu Leather Tanning Industrial District, located at the
western part of Turkey. The district houses more than 100
large tanneries with a production capacity representing a
major fraction of the sheepskin processing in Turkey.
2.2. Conventional characterization
Conventional characterization of the raw tannery wastewater
was carried out on a grab sample, collected from the
equalization tank of the WWTP of the above-mentioned
industrial district. The relevant analyses were performed in
duplicate in accordance with the Standard Methods (APHA
et al., 1998). COD, which was the key parameter in this study,
was also measured in duplicate according to the International
Standard ISO 6060 (International Organization for Standardi-
zation, 1986). For spectrophotometric color determination,
optical density values after filtering the raw wastewater
through 450 nm membrane (at original pH) were recorded at
three different dominant wavelengths of 436, 525 and 620 nm,
in accordance with the German Legislation (Anhang 38, 1993,
updated online in 2004). Total suspended solids (TSS) and
volatile suspended solids (VSS) measurements were con-
ducted after filtering the sample from Millipore AP40 glass
fiber filters with an effective pore size of approximately
1200–1600 nm.
2.3. Sequential filtration/ultrafiltration
A continuously stirred cell with a volumetric capacity of
400 mL (Amicon, Model 8400) was used as the filtration/
ultrafiltration unit, and was operated under positive pressure
(0.6–1.2 atm; N2 as the inert gas) during the experiments.
Operational parameters, such as temperature and working
pressure, applied during the sequential filtration/ultrafiltra-
tion experiments were the same as those reported in a
previous study by Dulekgurgen et al. (2006) and within the
ranges recommended by the manufacturers. No pH adjust-
ment was needed since the original pH of the raw wastewater
(Table 1) was within the range applicable for sequential
filtration/ultrafiltration experiments (Dulekgurgen et al.,
2006). Conventional disposable filters with pore sizes of
1200–1600 nm (Millipore AP40, glass fiber), 450 nm (Durapores
HV, polyvinylidene fluoride (PVDF)), and 220 nm (Durapores
GV, PVDF) (Millipore Corp., Bedford, MA 01730) were used for
sequential filtration. Permeate from the final filtration step
was successively passed through ultrafiltration membrane
discs with nominal molecular weight cut-off (MWCO) values
of 100, 30, 10, 3, and 1 kDa (PL series, Millipore, MA). In order
to provide consistency among the different size units of
ultrafiltration and filtration, the nominal MWCO values given
in kDa units were approximated to the corresponding particle
size values defined in nm, by using the approach described by
Cheryan (1986) and McGregor (1986). No interference due to
interaction between the filters and the tannery wastewater
 ARTICLE IN PRESS
WAT E R R E S E A R C H 42 (2008) 1083 – 1092 1085

Table 1 – Conventional wastewater characterization tions were added right before the start of respirometric test in
each case. The nutrient solutions were prepared in accor-
Parameters Ates et al. (1997) This dance to the recipes given by O’Connor (1972), and 10 mL from
study each was added per 1000 mg COD present in the sample.
Mean Range Nitrification inhibitor (Formula 2533–35, Hach Company) was
added in each case (0.16 g per 300 mL), in order to prevent any
COD total (mg L1) 4947 2513–8781 3100
interference from the nitrification process to the OUR
COD filtered through 1770 1284–3125 1240
1600 nm (mg L1)
assessments. Respirometric tests were performed using a
COD filtered through 450 nm – – 1195 PC-connected respirometer (Manotherm RA-1000). The respi-
(mg L1) rometer was inline with a 3-L aerated reactor and a 750-mL
TSS (mg L1) 2239 1000–4740 1940 measurement chamber, where the dissolved oxygen con-
VSS (mg L1) 1131 650–1540 1135 sumption in the mixed liquor was continuously monitored.
VSS/SS (%) 51 59 The aerated reactor was seeded with biomass obtained from
TKN (mg L1) 214 208–220 130
the WWTP, which was also sampled for the raw tannery
NH3-N (mg L1) 95 56–136 54
TP (mg L1) 9 3–22 13 wastewater investigated in this study. The endogenous
pH 8.4 6.4–10.1 8.3 respiration level of the sludge was determined at the
Alkalinity (mg CaCO3 L1) 665 259–1132 1010 beginning of each respirometric test. The amount of waste-
Chloride (mg Cl L1) 7601 6150–9060 4150 water/filtrate added was adjusted such that an average initial
Color (filtered through food-to-microorganism (F/M) ratio of 0.15 gCOD (gCOD)1 was
450 nm)
obtained in all runs. All the experiments were carried out at a
l ¼ 436 nm – – 0.178
constant temperature of 20 1C and aeration was supplied
l ¼ 525 nm – – 0.123
l ¼ 620 nm – – 0.090 continuously to maintain a dissolved oxygen concentration
above 6 mg L1, at all times. OUR data were collected online.
pH was 7.0–8.0 during all experiments owing to the buffering
capacity provided by the addition of the nutrient solutions.
The readily biodegradable COD (SS), like all other biode-
was expected since the filter materials were chemically gradable COD fractions, is calculated using the amount of
compatible with a wide range of solvents and were reported oxygen utilized, in other words the area under the OUR curve
to have no adsorptive capacity for soluble organics. To was used for the estimation of biodegradable COD. The
prevent fouling and other similar effects, the procedures procedure to define the readily biodegradable COD (SS) using
recommended by the manufacturer for cleaning, testing and respirometry was first suggested by Ekama et al. (1986). Its
conditioning of the stirred-cell and ultrafiltration membrane reliability was tested extensively and the procedure was
discs were performed. Moreover, to avoid any undesired improved by Cokgor et al. (1998). OUR data obtained for raw
filtering effect due to formation of a cake on top of the wastewater and filtrates were used in the modeling studies
filtering media and to avoid premature polarization, the for determination of the COD fractions and the kinetic and
height of the vortex formed during mixing was kept at 1/3rd stoichiometric coefficients of the applied mechanistic model.
of the total height of the sample column at all times, as Aquasim software (Reichert et al., 1998) was used to perform
recommended by the manufacturer. The special design of the the model simulations.
magnetic stirrer enabled continuous mixing in suspension
without touching the filter membranes/discs.
In addition to 100 mL permeate for COD measurements in 3. Results and discussion
duplicate, approximately 500 mL permeate of each filtering
step was secured for respirometric tests. Thus, filtration was 3.1. Conventional characterization
started with an initial volume of 5 L raw wastewater.
For a better interpretation of the results, the particle size Conventional characterization results for raw wastewater
values were grouped into operational size categories being taken from the equalization tank of the WWTP serving the
particulate (remaining above AP40 glass fiber filter, including Corlu Leather Tanning Industrial District is given in Table 1,
the settleable (4105 nm) and most of the supracolloidal together with the mean values and the ranges determined in
(103–105 nm) substances), colloidal (in the range of 2 nm a previous study (Ates et al., 1997), in which the same
(1 kDa membrane) to 1600 nm (AP40 glass fiber filter)) and wastewater source was monitored for more than a year. As
soluble (filtered through 1 kDa ultrafiltration membrane can be seen from the table, the investigated wastewater might
(o2 nm)) fractions. be classified as a strong wastewater in terms of total COD
content. Other parameters with significant values are nitro-
2.4. Respirometry and modeling gen species, alkalinity, chloride and pH. The sampled waste-
water is determined to be deficient in total phosphorus,
A series of respirometric analyses were conducted with the which is a typical trait for tannery wastewaters. Values of
raw tannery wastewater and with each aliquot collected from selected parameters determined in this study fall within the
consecutive filtration/ultrafiltration steps. To secure the range of the general characteristics outlined by the above-
nutritional requirements and to provide the necessary buffer mentioned, long-term survey, though they are closer to the
capacity during the respirometric tests, two nutrient solu- lower end. This difference might be attributed to the nature
 ARTICLE IN PRESS
1086 WAT E R R E S E A R C H 42 (2008) 1083– 1092

and timing of sampling in this study (grab sampling during
wet season), as well as to the seasonal process variations.
3.2. PSD-based COD fractionation
No quiescent settling was applied prior to the experiments,
thus the total COD value (3100 mg L1), reflects the sum of
settleable-, supracolloidal-, colloidal- and soluble-COD frac-
tions. The COD values measured at the aliquots collected
after each filtration step can be addressed as cumulative values
since each of them corresponds to the total COD below the
designated filter size. The difference between two succes-
sively measured COD values is defined as differential COD and
corresponds to the COD content originating only from the size
range specified by the two consecutive filter sizes. PSD-based
have a soluble rapidly hydrolysable COD fraction (SH) and a
particulate slowly hydrolysable COD fraction (XS) (Orhon
et al., 1999b, c).
As an initial step, model simulation and calibration of the
OUR profiles were carried out with ASM3 proposed by Gujer
et al. (2000) and with the simultaneous storage and direct growth
model proposed by Karahan et al. (2006). The simulation and
calibration exercise was first started with models previously
suggested in the literature, i.e., ASM3 and simultaneous storage
ASM3-The original model

COD fractionation of the raw tannery wastewater, in terms of 45

differential COD values, is visually presented in Fig. 1. OURdata
40
As apparent from the figure, the investigated wastewater
35 OURmodel.total
has a fairly simple COD fingerprint with two significant
portions at the two ends of size distribution—the particulate 30 OURend.dec.

OUR (mg L-1 h-1)

and the soluble ranges. 60% of the total COD originates from 25 OURstorage
the particulate fraction, while 25% comes from the soluble
20
portion. The remaining 15% is well distributed among the OURend.resp.Xsto
colloidal range. This fingerprint is predictable, since raw 15
OURgrowthXsto
tannery wastewaters are generally characterized with high 10
particulate matter (contributing to particulate COD fraction)
5
and high soluble organic content (contributing to soluble COD
fraction), both originating from the raw organic materials 0
(sheepskin, etc.) processed during production. 0 2 4 6
Time (h)

3.3. Respirometric analyses and modeling studies

ASM3-including simultaneous direct growth on
3.3.1. Conceptual framework for modeling and storage of SS

Tannery wastewaters have been characterized to include a 45

OURdata
readily biodegradable COD fraction (SS), which contains a 40
considerable amount of volatile fatty acids (VFAs). VFAs can OURmodel.total
35
be stored as poly-hydroxy-alkanoates by bacteria under OURend.dec.
30
OUR (mg L-1 h-1)

dynamic conditions and recent studies have shown that

OURstorage
substrate storage phenomena play an important role on the 25
OURend.resp.Xsto
biochemical transformations involved in the biological treat- 20
ment of tannery effluents (Dizdaroglu-Risvanoglu et al., 2007). OURgrowthXsto
15
It has been previously stated that tannery wastewaters also OURgrowthSS
10

2000 0
0 2 4 6
Differential COD (mg L-1)

Time (h)
1500
Fig. 2 – Simulation results for tannery wastewater (filtered
through 450 nm) with (a) the original ASM3 model and
1000
(b) simultaneous storage and direct growth model (Karahan
et al., 2006). OURdata: Experimental OUR data;
500 OURmodel.total: Total OUR estimated with the
corresponding model; OURend.dec: OUR estimated for
endogenous decay; OURstorage: OUR estimated for storage;
0
OURend.respXsto: OUR estimated for endogenous
>1

45

22

13

8-

5-

3-

2-

<2
13

3
0-

0-

-2
60

respiration of stored polymers; OURgrowthXsto: OUR

nm
nm

nm

nm
16

45

20
0

nm
nm

00

nm

estimated for growth on stored polymers; OURgrowthSs:

nm
nm

OUR estimated for direct growth on readily biodegradable

Fig. 1 – COD fingerprint of the raw tannery wastewater. substrate.
 ARTICLE IN PRESS
WAT E R R E S E A R C H 42 (2008) 1083 – 1092 1087

and direct growth model proposed by Karahan et al. (2006).
The calibration approach involved the use of calculated COD
fractions using the process yields reported by Dizdaroglu-
Risvanoglu et al. (2007) for a similar tannery wastewater from
a different district. Model calibrations were carried out in an
iterative manner, i.e. a change in one parameter for the best
fit of a particular OUR curve was applied to the simulations
performed for all six different OUR profiles. However, the
results of the best fits obtained with both models were not
able to describe the observed OUR curves with two local
maxima, indicating two distinct biochemical processes
(Fig. 2).
Although it was not the initial intention of this study, an
amended mechanistic model was designed by necessity, in
order to simulate the biodegradation of different COD
fractions of the tannery effluent under investigation. The
matrix presentation of the amended model is given in Table 2.
The schematic representation of the conversions of different
COD fraction in view of the proposed model structure is given
in Fig. 3. The first row in the table includes the model
components and the last column defines the rates for each
process involved in the model. The amended model suggests
that the readily biodegradable COD fraction (SS) is utilized to
be converted to storage polymers (XSTO). It is assumed that SS
fraction is first converted to the storage polymers, and growth
on these polymers starts after the total depletion of all readily
biodegradable COD in the bulk solution. This sequence is
obtained with the use of the switch function KS/(KS+SS) in the
process rate expression of growth on XSTO as given in Table 2.
The model also involves two hydrolysis processes for rapidly
and slowly hydrolysable COD fractions (SH and XS). Hydrolysis
products of both processes are labeled as hydrolyzed easily
biodegradable COD (SShyd) and this portion of COD is directly
used for biomass growth according to the model. This
assumption is acceptable based on the fact that hydrolysis
processes are rate-limiting processes and generate easily
utilizable products at moderate rates, which cause the
activated sludge culture to establish a balanced growth
mechanism (Karahan et al., 2006). The endogenous decay of
heterotrophic biomass and the decay of storage polymers in
the model are defined as similar to those proposed in ASM3
(Gujer et al., 2000).

Table 2 – Matrix presentation of the amended model for degradation of tannery wastewater

Component process SO SI SS SH SShyd XI XS XH XSTO Rate

O2 COD COD COD COD COD COD COD COD

Rapid hydrolysis 1 1 kSH

ðSH =XH Þ
XH
½KSH þðSH =XH Þ
Slow hydrolysis 1 1 ðXS =XH Þ
kXS ½K þðX XH
XS S =XH Þ
Aerobic storage of SS ð1YSTO Þ 1/YSTO 1 SS SO
 YSTO kSTO K XH
S þSS KO þSO
Direct growth on SShyd ð1Y Þ 1/YH1 1 SShyd SO
 Y H1 mH1 K XH
H1 Shyd þSShyd KO þSO

Growth on XSTO 
ð1YH2 Þ 1 1/YH2 mH2 ½K
ðXSTO =XH Þ SO KS
XH
YH2
STO þðXSTO =XH Þ KO þSO KS þSS
Endogenous respiration (1fSIfI) fSI fI 1 SO
bH K þS XH
O O
Respiration of XSTO 1 1 SO
bSTO K XSTO
O þSO

Fig. 3 – Schematic representation for conversion processes of different COD fractions.

 ARTICLE IN PRESS
1088 WAT E R R E S E A R C H 42 (2008) 1083– 1092

As shown in Fig. 4, this approach enables successful model aliquot from 2 nm filter and was carried out for bigger pore
simulation of the two distinctive OUR responses obtained in sizes considering that the same COD fractions have to be
the system. present in those permeates. The obtained COD fractions were
then used as data for model simulations, which gave
3.3.2. Process stoichiometry and kinetics for tannery effluents consistent results with the same parameter set for all six
Values for storage yield (YSTO), heterotrophic yield for direct OUR profiles. Thus, the stoichiometric estimations of COD
growth (YH1), and secondary heterotrophic growth yield on fractions were justified by model simulations.
stored material (YH2) used for simulations in this study were The readily biodegradable fraction (SS) of tannery effluent
chosen as 0.83, 0.68, and 0.79 gCOD (gCOD)1, respectively. was found as 230 mg L1 corresponding to 7% of the total COD.
Instead of using default ASM3 values, the above-mentioned The rapidly hydrolysable fraction (SH) was 615 mg L1 (20%),
stoichiometric values were adopted from the study by and the soluble inert fraction (SI) was 395 mg L1 (13%). The
Dizdaroglu-Risvanoglu et al. (2007), which was conducted on value of soluble inert COD (SI) was found to be 13% of total
tannery effluents from a different source (Tuzla Organized COD and this result is in agreement with the reported range
District of Tannery Industries). Model calibration results for of 9–14% (Orhon et al., 1999d).
raw wastewater and filtered samples are summarized in
Table 3. Values of the kinetic coefficients obtained by model
simulations given in Table 3 also agree with those previously
reported for tannery wastewaters by Dizdaroglu-Risvanoglu
Table 3 – Kinetic coefficients determined for raw and
et al. (2007). The simulation results for six respirometric tests
filtered wastewater samples
conducted with wastewater samples obtained from succes-
sive filtration/ultrafiltration are shown in Fig. 5. The simula-
tion result for the wastewater sample filtered through 450 nm Kinetic Units Filtered Raw
coefficients samples sample
was already given in Fig. 4.
bH d1 0.10 0.08
3.3.3. COD fractions based on PSD bSTO d1 0.10 0.08
Model simulation and calibration of OUR profiles require kSTO d1 3 3
correct assessment of the COD fractions as respective model KS mg COD L1 12 12
kSH d1 2 1.8
components. Accordingly, the overall COD fractions esti-
KSH gCOD (gCOD)1 0.01 0.01
mated by combining physical segregation based PSD and
kXS d1 – 1.7
biodegradability based modeling studies are given in Table 4. KXS gCOD (gCOD)1 – 0.05
This estimation procedure involves the calculation of biode- mH1 d1 2 1.8
gradable COD for each OUR profile obtained for each KShyd mg COD L1 20 20
permeate, as described by Karahan et al. (2002). The mH2 d1 2 2
procedure was started by calculating the COD fractions for KSTO gCOD (gCOD)1 0.2 0.2

ASM3-including dual hydrolysis, storage of SS and direct

growth on hydrolysis products
45
OURdata
40
OURmodel.total
35 OURend.dec.
OUR (mg L-1 h-1)

30 OURstorage
OURend.resp.Xsto
25
OURgrowthXsto
20
OURgrowthSShyd
15

10

0
0 2 4 6 8 10 12 14
Time (h)

Fig. 4 – Simulation results for tannery wastewater (filtered through 450 nm) with ASM3 amended to include dual hydrolysis
and direct growth on hydrolysis products. OURdata: Experimental OUR data; OURmodel.total: Total OUR estimated with the
adopted model; OURend.dec: OUR estimated for endogenous decay; OURstorage: OUR estimated for storage;
OURend.respXsto: OUR estimated for endogenous respiration of stored polymers; OURgrowthXsto: OUR estimated for growth
on stored polymers; OURgrowthSshyd: OUR estimated for direct growth on hydrolyzed slowly biodegradable substrate.
 ARTICLE IN PRESS
WAT E R R E S E A R C H 42 (2008) 1083 – 1092 1089

Part icle Size < 3 nm Part icle Size < 5 nm

30 30
OURdata OURdata
25 OURmodel 25 OURmodel
OUR (mg L -1 h -1)

OUR (mg L -1 h -1)

20 20

15 15

10 10

5 5

0 0
0 2 4 6 8 10 12 14 0 2 4 6 8 10 12 14
Ti me (h) Ti me (h)

Part icle Size < 8 nm Part icle Size < 13 nm

30 35
OURdata OURdata
25 OURmodel 30 OURmodel
25
OUR (mg L -1 h -1)

OUR (mg L -1 h -1)

20
20
15
15
10
10

5 5

0 0
0 2 4 6 8 10 12 14 0 2 4 6 8 10 12 14
Ti me (h) Ti me (h)
Part icle Size < 1600 nm
40

35 OURdata
OURmodel
30
OUR (mg L -1 h -1)

25

20

15

10

0
0 2 4 6 8 10 12 14
Ti me (h)

Fig. 5 – Simulation results obtained for filtrates collected after each filtration/ultrafiltration step.

Calibration of the OUR profiles of the filtrates after each of 75% was predicted to be involved in the calculation of
filtration/ultrafiltration step provided, as given in Table 4, biodegradable COD fractions, corresponding to a maximum
related COD fractionation for all major soluble components. error of 15 mgCOD L1 for SS, 30 mgCOD L1 for SH and
The value of each COD component also indicated the 15 mgCOD L1 for XS, which would be acceptable in terms of
cumulative level associated with all the lower sizes. This the scope of this study. Both COD fractions and modeling
way, the adopted approach enabled verification of applicable were performed using the same yield coefficients.
COD fractionation by model simulation and yielded a particle As given in Table 4, the COD fraction referred to as the
size distribution of all major COD fractions. The confidence readily biodegradable fraction (SS) was entirely at the soluble
intervals related to the calculation of COD fractions were range (o2 nm). The majority (83%) of the COD portion defined
determined by the experimental errors involved in COD as the rapidly hydrolysable fraction (SH) and accepted as
measurements (75 mgCOD L1) and those of OUR measure- soluble in the model was actually at the soluble range, and
ments (70.5 mg OUR L1 h1). In this context, an overall error only a minor portion (17%) of SH was at the colloidal range
 ARTICLE IN PRESS
1090 WAT E R R E S E A R C H 42 (2008) 1083– 1092

(13–220 nm). Interestingly, only 11% of the COD fraction

1860
CTb

220

780
45

95
25
65
5

5
addressed as the soluble inert fraction (SI) in the model was
at the soluble range, whereas the rest (89%) was determined

240

105

740
to be distributed over the entire colloidal range (2–1600 nm).
CS
Table 4 – Size distribution of the COD fractions of the tannery wastewater obtained by sequential filtration/ultrafiltration, respirometric analyses and modeling

0
0

0
0
0
0
The particle size distribution mentioned above is found to be
Differential COD (mg L1)

quite wastewater specific, as it differs, for example, from that

1620
XI

0
0

0
0
0
0
0
0
of a textile wastewater where the majority of SI was located
below 2 nm (Dulekgurgen et al., 2006).
The area under the OUR curve obtained in the respirometric
240
XS

0
0

0
0
0
0
0
0
test conducted with raw tannery wastewater (Fig. 6) can be
used to estimate the total amount of biodegradable substrate
115
45

95
25
65
40
SI

(CS) utilized during the test. The evaluation initially presumed

0

that all biodegradable substrate was consumed during the

test. The amount of dissolved oxygen consumed was
105

510
SH

0
0
0

0
0
0
0

relatively low and could only be justified with an overall yield

coefficient of 0.79 gCOD (gCOD)1. However, the repeated
230
SS

0
0
0

0
0
0
0
0

evaluation indicated that around 20–22% of the slowly

biodegradable COD remained unutilized after the test re-
adjusting the yield coefficient to 0.64–0.66 gCOD (gCOD)1,
Size category (nm)

which compares well with the adopted values for other tests.
Accordingly, the CS fraction was estimated as 1085 mg L1
450–1600
220–450

13–220
41600

8–13
5–8
3–5
2–3
o2

with an average net yield value of 0.79 gCOD (gCOD)1, the

slowly hydrolysable COD fraction (XS) was found as
240 mg L1, and the particulate inert fraction (XI) as
1620 mg L1. In the literature, XI was reported to range
between 7% and 19% of the initial total COD content of
3100

1240
1195
1190

tannery wastewater samples (Orhon et al., 1999d). Comparing

CTa

970
965
870
845
780

the values for XI and XS obtained via modeling in this study to

those given in the literature, it is apparent that biodegrada-
1085

845
845
845

740
740
740
740
740

tion of XS was hindered, and a significant portion of that COD

CS

fraction appeared to be inert. Calibration of the model using

Cumulative COD (mg L1)

the respirometric data of the raw wastewater also revealed

1620
XI

rapid hydrolysis and direct growth rates being 10% lower than
0
0
0

0
0
0
0
0

the values obtained for the filtered portions (Table 3). The
endogenous decay of heterotrophic biomass and stored
240
XS

0
0
0

0
0
0
0
0

polymers were also 20% slower for the biomass fed with
raw wastewater sample. In fact, a 10–20% inhibition is, in
395

395
350
345

230
225
130
105

general, not so important, but when reflected on the OUR

42
SI

profile as illustrated in Fig. 6, it induces a significant

difference which may be interpreted in terms of rates of
CT: differential COD values (at each size category) seen in Fig. 1.
CT: COD measured after each filtration step (cumulative values).
615

615
615
615

510
510
510
510
510
SH

different processes.
These results showed that the particulate portion of the
230

230
230
230

230
230
230
230
230

tannery effluent imposed inhibitory effects on the biochem-

SS

ical reactions and confirmed the use of respirometry as the

most useful tool for the assessment of inhibitory effects as
Particle size (nm)

suggested in previous studies (Insel et al., 2006). This

1200–1600

inhibition was observed as a decrease in the amount of

450
220

13

biodegradable COD (CS), namely only 20% of the slowly

8
5
3
2

hydrolysable COD (XS) could be degraded during the test.

Inhibition was also detected as a decrease in the rates of
hydrolysis, growth and decay related processes, indicating a
non-competitive inhibition where a portion of the available
Separation technique

enzymes is irreversibly blocked by the inhibitor. This inhibi-

tion, among other possible complex organic compounds, can
be mostly attributed to the presence of trivalent chromium,
Ultrafiltration

which is reported to be of particulate nature, probably

AP40 filter
HV filter
GV filter

entrapped in the particulate organic matter (Ates et al.,

100 kDa
Filtration

30 kDa
10 kDa
3 kDa
1 kDa

1997). These inhibitory effects can clearly be seen in Fig. 6,

Total

where OUR responses of the system were simulated for

b
a

both the real case, when the process is inhibited, and for the
 ARTICLE IN PRESS
WAT E R R E S E A R C H
Verification of Inhibition via Modeling
45
40
35
OUR (mg L -1 h -1)
30
25
20
15
10
0
0 2
Fig. 6 – Simulation result obtained for raw wastewater.
non-inhibitory conditions, when no inhibition was imposed
on the system, namely using the same set of parameters
obtained for the filtered effluents. The results on the raw
wastewater have been included in this study for comparison
only and due to the merit of the filtration/ultrafiltration,
which practically removes and eliminates inhibitors beyond a
given size range, general biodegradation kinetics could be
applied.
4. Conclusions
In the light of the results presented in the preceding sections,
the concluding remarks of the study may be outlined as
follows:
(i) The particle size analysis yielded a specific COD finger-
print for the tannery wastewater, with two significant
portions at the two ends of size distribution: 60% of the
total COD was at the particulate range, 25% was at the
soluble range and the remaining 15% was well distributed
among the colloidal range.
(ii) The results also showed that respirometry could be used
as a useful complement of particle size distribution by
ultrafiltration for the assessment of biodegradation
characteristics of COD fractions within each size interval.
The OUR profiles associated with different size ranges
between 2 and 1600 nm could be calibrated with the same
set of stoichiometric and kinetic coefficients. Calibration
also verified and confirmed the validity of the amended
model defined as a modified version of ASM3, including
direct growth on hydrolysis products.
(iii) The combined analysis of filtration/ultrafiltration and
OUR modeling yielded not only the particle size distribu-
tion of all major COD fractions, but also the extent of
their biodegradability. In particular, this approach indi-
cated that, while the majority of SH was in the soluble
range (o 2 nm) for the tannery wastewater selected in the
study, SI was well distributed among the colloidal range
42 (2008) 1083 – 1092 1091
OURdata
OURmodel
(inhibited)
OURmodel
(no inhibition)
4 6 8 10 12 14
Time (h)
and therefore removable by physical entrapment and
adsorption.
(iv) Joint evaluation of particle size distribution and respiro-
metric analysis of COD fractions for significant size
intervals offers a significant potential, as demonstrated
for tannery wastewater in this study, for providing a new
insight for the biodegradation pattern of complex waste-
waters. Similar studies are recommended for different
wastewaters and also on process effluents, particularly
for the assessment and particle size distribution of
residual soluble microbial products.
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