Bioresource Technology 121 (2012) 31–35

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Antibiotic Fermentation Broth Treatment by a pilot upflow anaerobic sludge

bed reactor and kinetic modeling
T. Coskun a,⇑, H.A. Kabuk a, K.B. Varinca a, E. Debik a, I. Durak b, C. Kavurt c
a
Environmental Engineering Department, Yildiz Technical University, 34220 Esenler, Istanbul, Turkey
b
Deva Kartepe API Production Plant, 41135 Kocaeli, Turkey
c
Hidromer Aritma Sistemleri Ltd. Sti., 41040 Kocaeli, Turkey

h i g h l i g h t s

" Anaerobic fermentation broth could be efficiently treated by upflow anaerobic sludge blanket (UASB) reactor.
" The highest chemical oxygen demand (COD) removal efficiency was obtained as 95.7%.
" The highest methane production rate was 0:30 Lmethane g1
CODremoved .
" The results of the study were interpreted using the various kinetic models.
" The results showed a close relationship with all kinetic models used.

a r t i c l e i n f o a b s t r a c t

Article history: In this study, an upflow anaerobic sludge blanket (UASB) mesophilic reactor was used to remove antibi-
Received 17 May 2012 otic fermentation broth wastewater. The hydraulic retention time was held constant at 13.3 days. The
Received in revised form 27 June 2012 volumetric organic loading value increased from 0.33 to 7.43 kgCOD m3 d1 using antibiotic fermentation
Accepted 28 June 2012
broth wastewater gradually diluted with various ratios of domestic wastewater. A COD removal effi-
Available online 7 July 2012
ciency of 95.7% was obtained with a maximum yield of 3,700 L d1 methane gas production. The results
of the study were interpreted using the modified Stover–Kincannon, first-order, substrate mass balance
Keywords:
and Van der Meer and Heertjes kinetic models. The obtained kinetic coefficients showed that antibiotic
Anaerobic treatment
Antibiotic fermentation broth wastewater
fermentation broth wastewater can be successfully treated using a UASB reactor while taking COD
Kinetic model removal and methane production into account.
UASB Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction are required. Upflow anaerobic sludge blanket (UASB) reactors

Antibiotic production is a significant portion of the pharmaceu-
tical industry. In the past, antibiotics were obtained from a variety
of bacteria and fungi. Currently, antibiotics can be produced from
natural or synthetic chemicals. One of the most important methods
of antibiotic production is passing fermentation broth containing
clavulanic acid through membrane systems to separate out impu-
rities (Brites Alves et al., 2002). The portion that fails to pass the
ultrafiltration membrane is referred to as the waste of the fermen-
tation broth. A review of the literature indicates that there is no
study regarding the disposal of these wastes, and they pose impor-
tant problems for the industries producing the raw materials for
antibiotics in Turkey. These wastes cause damage to the natural
environment, due to the content of hardly biodegradable organic
materials. For this reason, studies on the treatment of these wastes
have not been used in the pharmaceutical industry to treat this
type of waste, even though they have been successfully used for
other types of wastewater (Chen et al., 2011; Guo et al., 2012; Ok-
tem et al., 2008; Satyanarayan et al., 2009). In this study, the treat-
ment of antibiotic fermentation broth waste was examined using a
UASB reactor. The results of this study were applied to the modi-
fied Stover–Kincannon, first-order, substrate mass balance and
Van der Meer and Heertjes kinetic models, and the kinetic coeffi-
cients obtained in this study were compared with those found in
the literature.
2. Methods
2.1. Anaerobic digester

⇑ Corresponding author. Tel.: +90 2123835379; fax: +90 2123835356. A pilot UASB reactor with a volume of approximately 2 m3 was
E-mail address: tcoskun@yildiz.edu.tr (T. Coskun). used in this study. Wastewater was fed to the reactor on a daily ba-

0960-8524/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.06.102
32 T. Coskun et al. / Bioresource Technology 121 (2012) 31–35
sis using a pump. There were two level sensors in the reactor set to
the minimum and maximum water levels. There was a discharge
valve that was automatically opened when the water reached the
maximum level and closed when the water reached the minimum
level. There was one additional manual valve to discharge the
sludge when the sludge level rose. The biogas production in the
reactor was continuously measured by a gas meter and the meth-
ane production values were calculated as regularly to determine
the methane content in the biogas by a gas analyzer (Geotec,
GA2000). The study results were evaluated according to the meth-
ane values for the kinetic models.
2.2. Wastewater
The wastewater used in this study was taken from an antibiotic
producing facility in which fermentation broth containing clavu-
lanic acid is passed through an ultrafiltration membrane to sepa-
rate out colloidal particles and impurities. The waste broth used
in this study was the portion that did not pass through the ultrafil-
tration membrane. Waste broth was mixed with domestic waste-
water for dilution before feeding the reactor to obtain the
desired volumetric organic loading rate value. A low volumetric or-
ganic loading rate was maintained during the acclimatization per-
iod. Next, the volumetric organic loading rate was increased
stepwise according to the chemical oxygen demand (COD) removal
efficiency and biogas production. The characteristics of the waste
broth are shown in Table 1. All of the analyses were conducted
according to standard methods (APHA, 2005).
2.3. Operating conditions
One hundred and fifty liters of wastewater was fed to the UASB
reactor daily to allow for a hydraulic retention time of approximately
13 days. Broth diluted with wastewater was fed into the reactor. The
organic loading rate was increased by reducing the dilution by waste-
water after the desired efficiency values were obtained. The values of
the organic loading rates ranged from 0.33 to 7.43 kgCOD m3 d1. The
reactor temperature was held constant at 35 °C by passing hot water
through the heat jacket surrounding the reactor.
2.4. Kinetic models
Mathematical models are used for the prediction of outcomes as
a result of various inputs. Mathematical models are also used to
determine system performance and optimal operating conditions.
Although there are various models developed for anaerobic treat-
ment, depending on the type of wastewater and operating condi-
tions, each kinetic model is only suitable for specific cases and
processes. The suitability of the results of this study was deter-
mined according to the modified Stover-Kincannon, substrate mass
balance, first-order and Van der Meer and Heertjes models. Next,
the kinetic coefficients obtained from models were compared to
values obtained in the literature, as shown in Table 2.
2.4.1. Modified Stover–Kincannon model
The original Stover–Kincannon model was carried out on a
rotary biodisc. For rotating biodiscs, suspended solids can be
Table 1
Waste fermentation broth characteristics.
Parameter Unit Value
pH 5.5 ± 0.5
Solid materials % 7.0 ± 1.0
Crude protein % 2.6 ± 0.5
Crude fats % 2.5 ± 0.5
COD g L1 150.0 ± 10.0
neglected due to the addition of biomass with the biofilm surface
area considered for the definition of biomass (Kincannon and Sto-
ver, 1982). On the other hand, biomass has been expressed by the
volume in the modified model (Yu et al., 1998). The substrate con-
sumption rate is explained as a function of the volumetric organic
loading in the modified Stover–Kincannon model (Isik and Sponza,
2005). Reactions for the modified Stover–Kincannon model are de-
scribed below (Yu et al., 1998):
dS Q
¼ ðSi  Se Þ ð1Þ
dt V
In this equation, Q is the wastewater flow (L d1), V is the vol-
ume of the reactor (L), Si is the input COD concentration and Se rep-
resents the concentration of the output COD. The dS/dt expression
in Eq. (1) can be explained as follows:
dS U max ðQSi =VÞ
¼ ð2Þ
dt K B þ ðQSi =VÞ
In this equation, Umax is the maximum substrate consumption
rate (g L1 d1), and KB is the constant saturation value (g L1 d1).
The new equation, combining Eqs. (1) and (2), can be written as
follows:
Q U max ðQSi=VÞ
ðSi  SeÞ ¼ ð3Þ
V K B þ ðQSi=VÞ
If both sides of the equation are reversed and rearranged, the
equation becomes
V KB V 1
¼ þ ð4Þ
QðSi  Se Þ U max QSi U max
According to Eq. (4), if a graph is drawn with V/QSi values corre-
sponding to the values of V/Q(SiSe), the slope of the line provides
the KB/Umax value. The point at which the line crosses the Y-axis
provides the value of 1/Umax.
2.4.2. First-order kinetic model
The first-order model refers to substrate removal in the system.
Accordingly, the equation is expressed as follows:
dS Q
 ¼ ðSi  Se Þ  kSe ð5Þ
dt V
In this equation, k refers to the first-order kinetic coefficient
(d1). For steady-state conditions, dS/dt is 0. Accordingly, this equa-
tion becomes
ðSi  Se Þ
¼ kSe ð6Þ
h
According to Eq. (6), if a graph is drawn for Se values corre-
sponding to the (SiSe)/h values, the slope of the line will provide
the k kinetic coefficient.
2.4.3. Substrate mass balance model
In an anaerobic reactor, while part of the organic matter is leav-
ing the system with the output water, the remainder passes
through a portion of the structure of the new cell, and a section
is used for energy. Biogas is released as a result of fragmenting
the organic matter for energy. If a mass balance equation was made
for the substrate, the following is obtained (Raposo et al., 2004):
ðTCODÞ0 ¼ ðSCODÞeffluent þ ðTCODÞm þ ðTCODÞVSSe
þ ðTCODÞbiogas ð7Þ
In this equation, (TCOD)0 indicates a portion of the total COD of
the input wastewater, (SCODE)effluent indicates the portion that re-
sults from the dissolved COD of the output wastewater, (TCOD)m
indicates the portion of COD consumption caused by the activities
 T. Coskun et al. / Bioresource Technology 121 (2012) 31–35 33

Table 2
The comparison of the kinetic constants for the models used in this study.

Models Wastewater type OLR Reactor T (°C) Kinetic References

(gCOD L1 d1) type parameters
Umax KB
Modified Stover–Kincannon Distillery spent wash 5.0–20.0 BFFR 37 2.0 1.69 Acharya et al. (2011)
model Starch wastewater 1.15–16.70 AF 35 49.8 50.6 Ahn and Forster (2000)
Starch wastewater 1.15-.16.70 AF 55 667 702 Ahn and Forster (2000)
Municipal wastewater 1.2–11.7 UASB 17.1– 2.0 1.5 Aydinol et al. (2011)
21.0
Synthetic wastewater 1.0–10.0 AHR 35 83.3 186.2 Buyukkamaci and Filibeli
(2002)
Poultry slaughterhouse 0.64–4.97 SGBR at 164.5 177.2 Debik and Coskun (2009)
wastewater
Simulated textile wastewater 1.01–16.86 UASB 37 7.5 8.2 Isik and Sponza (2005)
Dyestaff 2.0–8.0 APCR 20 13.0 37.7 Kapdan (2005)
Saline synthetic wastewater 2.3–7.7 UAPB 37 5.3 7.1 Kapdan and Erten (2007)
Pig fattening farm liquid manure 0.32–5.25 ABBR 38 80.9 91.6 Kosinska and Miskievwichz
(2009)
Synthetic wastewater 0.31–3.25 AMBR – 29.5 31.6 Kuscu and Sponza (2009)
Formaldehyde containing 0.18–3.16 UAFB 30 3.4 4.6 Raja Priya et al. (2009)
wastewater
Textile wastewater 1.04–8.21 UAFB 30 31.7 45.4 Sandhya and Swaminathan
(2006)
Food processing wastewater 1.1–5.0 MACR 35 22.9 23.6 Senturk et al. (2010)
Milk permeate 2.0–20.0 AMBBR 35 89.3 102.3 Wang et al. (2009)
Papermill wastewater 1.07–12.25 AF 35 86.2 104.2 Yilmaz et al. (2008)
Papermill wastewater 1.07–12.25 AF 55 666.7 843.9 Yilmaz et al. (2008)
Soybean wastewater 3.7–24.5 AF 35 83.3 85.5 Yu et al. (1998)
Antibiotic fermentation broth 0.33–7.43 UASB 35 399.0 445.5 This study
YG/S km
Substrate mass balance model Food processing wastewater 1.1–5.0 MACR 35 0.394 0.02 Senturk et al. (2010)
Milk permeate 2.0–20.0 AMBBR 35 0.341 0.03 Wang et al. (2009)
Antibiotic fermentation broth 0.33–7.43 UASB 35 0.322 0.01 This study
ksg
Van der Meer and Heertjes Synthetic wastewater 0.31–3.25 AMBR – 0.095 Kuscu and Sponza (2009)
model Antibiotic fermentation broth 0.33–7.43 UASB 35 0.297 This study
k
First-order kinetic model Simulated textile wastewater 1.01–16.86 UASB 37 0.615 Isik and Sponza (2005)
Antibiotic fermentation broth 0.33–7.43 UASB 35 0.448 This study

at: ambient temperature; ABBR: Anaerobic bioreactor with biomass recycle; AF: Anaerobic filter; AHR: Anaerobic hybrid reactor; AMBBR: Anaerobic moving bed biofilm
reactor; AMBR: Anaerobic migrating blanket reactor; APCR: Anaerobic packed column reactor; BFFR: Biphasic fixed film bioreactor; MACR: High-rate mesophilic anaerobic
contact reactor; SGBR: Static granular bed reactor; UAFB: Upflow anaerobic fixed film reactor; UAPB: Upflow anaerobic packed bed reactor.

of microorganisms, (TCOD)VSSe indicates that portion of the total
particulate COD from the effluent, and (TCOD)biogas indicates the
produced biogas equivalent to the concentration of the total COD.
If the equation is edited, it can be expressed as
QSto ¼ QSse þ km XV þ Q ðSte  Sse Þ þ Q CH4 Y S=G
In this equation, Sto refers to the total COD concentration of the
input (gCOD L1), Sse is the dissolved COD concentration of the out-
put (gCOD L1), km is the new cell production kinetic coefficient
(d1), X is the concentration of microorganisms in the reactor
(gVSS L1), V is the reactor volume (L), Ste is the dissolved COD con-
centration of output (gCOD L1), QCH4 is the amount of methane pro-
duced (L d1), and YS/G is the ratio of substrate conversion to
methane (g COD g 1methane ). If Eq. (8) is rearranged, it can be expressed
as
QSto  QSte ¼ km XV þ Q CH4 Y S=G
To obtain a linear equation, both sides of Eq. (9) can be divided
by XV as follows:
Q ðSto  Ste Þ Q CH4
¼ km þ Y S=G
XV XV
If a graph is drawn according to Eq. (10), with Q CH4 =XV values
corresponding to Q (StoSte)/XV values, the slope of the line gives
the YS/G kinetic coefficient. The point at which the line crosses
the Y-axis provides the kinetic coefficient, km. If YS/G is reversed,
YG/S is obtained, which provides the amount of methane produced
per COD removal.
2.4.4. Van der Meer and Heertjes model
ð8Þ
Van der Meer and Heertjes developed a model for estimating
the production of methane gas (Van der Meer and Heertjes,
1983). In this model, produced methane per organic matter re-
moved is proportional to the kinetic coefficient of the Van der Meer
and Heertjes model, as indicated by ksg (Kuscu and Sponza, 2009):
Q CH4 ¼ ksg Q ðSi  Se Þ ð11Þ
In this equation, ksg refers to the kinetic coefficient of the Van
der Meer and Heertjes model (Lmethane g1 CODrem ).
3. Results and discussion
ð9Þ
3.1. COD removal
The organic loading rate gradually increased from 0.33 to
ð10Þ 7.43 kg m3 d1. The changes in the COD removal efficiencies cor-
responding to the organic loading rate are shown in Fig. 1.
Although the COD removal efficiencies decreased slightly when
the organic loading rate was increased, they remained above 80%
in general. During the study, the highest COD removal efficiency
34 T. Coskun et al. / Bioresource Technology 121 (2012) 31–35

100 10 between methane production and organic loading rate. During the
study, high methane production values (3,700 L) were obtained daily
COD removal efficiency, %

80 8 for both organic loading values of 6.6 kg m3 d1 and 7.4 kg m3 d1.

OLR, kg/m3-d
60 6 3.3. Kinetic modeling

40 4
3.3.1. Modified Stover–Kincannon model
The results of the modified Stover–Kincannon model are shown in
Fig. 3. As shown in Fig. 3, the study results were closely related to the
20 2
COD removal efficiency modified Stover–Kincannon model (R2 = 0.9938). The values of Umax
Organic loading rate and Kb were calculated as 399.0 g L1 d1 and 445.5 g L1 d1, respec-
0 0 tively. The kinetic coefficients determined in the literature are shown
0 20 40 60 80 in Table 2. Table 2 contains the results of studies in which different
Time, days wastewater and reactors were used and provides important infer-
ences from the results. Table 2 shows that temperature affected the
Fig. 1. COD removal efficiencies vs. organic loading rates. values of Umax and Kb significantly. However, there is a significant par-
allelism between kinetic coefficients Kb and Umax in all of the studies.
The Umax and Kb values obtained in this study are considerably higher
5000 10
than those from studies performed under mesophilic conditions. The
Umax and Kb kinetic coefficients show the biodegradability of organic
Methane production, L/d

4000 8 matter at current conditions. Thus, the obtained kinetic coefficients

show that antibiotic fermentation broth could be successfully treated
OLR, kg/m3 -d

3000 6 in this reactor type and at this temperature.

2000 4 3.3.2. First-order model

The first-order model was also used to interpret the study re-
1000 2 sults. According to Fig. 3, the results showed a close relationship
Methane production with the first-order model (R2 = 0.8010) but a less significant rela-
Organic loading rate tionship with the modified Stover–Kincannon model. Using Fig. 3,
0 0
0 20 40 60 80 the kinetic coefficient k was found to be 0.448 d1. Only a small
number of studies in the literature have used the first-order model.
Time, days
In one of these studies, the kinetic coefficient k was found to be
Fig. 2. Methane production vs. organic loading rates. 0.615 d1(Isik and Sponza, 2005). The first-order kinetic model
essentially reveals the change in efficiency due to changes in
obtained was 95.7%, while the COD removal efficiency obtained hydraulic retention time. Although the hydraulic retention time
with the highest organic loading rate (7.43 kg m3 d1) was 87.4%. was constant in this study, the results were still reasonably repre-
sented by the first-order kinetic model.
3.2. Methane production
3.3.3. Substrate mass balance model
Methane production increased with increasing organic loading The substrate mass balance model results are shown in Fig. 3.
rate during the study (Fig. 2). There is a strong relationship Fig. 3 shows that the results are closely related with the substrate

4 8000
y = 1,1165x + 0,0025 y = 0,4481x
V/(Q(Si-Se))

3 6000 2
2
R = 0,9938 R = 0,801
(Si-S)/T

2 4000

1 2000
a b
0 0
0 0,5 1 1,5 2 2,5 3 0 4000 8000 12000 16000
1/OLR S
0,30 4000
(Si-Se)/(HRT X)

y = 3,1009x + 0,01 y = 0,2971x

3000
0,20 R2 = 0,9336 R2 = 0,9375
QCH4

2000
0,10
1000
c d
0,00 0
0 0,02 0,04 0,06 0,08 0 2500 5000 7500 10000 12500 15000
QCH4/(XV) Q(Si-Se)

Fig. 3. Model results, (a) modified Stover–Kincannon (b) first order (c) substrate mass balance (d) Van der Meer and Heertjes).
 T. Coskun et al. / Bioresource Technology 121 (2012) 31–35
mass balance model (R2 = 0.9336). From this figure, YS/G and Km
were determined to be 3:101 g COD g 1
methane and
1
0:01 g CODremoved g 1
VSS d , respectively, while YG/S was determined
to be 0:322 g methane g 1 COD by taking the inverse of YS/G. According
to Table 2, YG/S had been obtained as 0:394 g methane g 1
COD (Senturk
et al., 2010) and 0:341 g methane g COD (Wang et al., 2009) in previous
studies. Thus, the 0:322 g methane g 1 COD value obtained in this study
was suitable with regards to the literature. Likewise, in this study,
1
the Km value of 0:01 g CODremoved gVSS1 d was close to those re-
ported in the literature.
3.3.4. Van der Meer and Heertjes model
The results for the Van der Meer and Heertjes model are shown
in Fig. 3. As shown in Fig. 3, the study results were closely related
with the Van der Meer and Heertjes model (R2 = 0.9375). From this
figure, the kinetic coefficient ksg was found to be
0:297 Lmethane g1
CODremoved for this study. Only a limited number of
studies have used the Van der Meer and Heertjes model. According
to the available literature, the kinetic coefficient ksg is given as
0:095 Lmethane g1
CODremoved for a study performed with synthetic
wastewater (Kuscu and Sponza, 2009). The Van der Meer and
Heertjes kinetic model reveals the amount of methane that is com-
posed of fixed organic matter. The value of 0:297 Lmethane g1
CODremoved
obtained in this study indicates that treating antibiotic fermenta-
tion broth wastewater anaerobically allowed a significant portion
of organic matter to change into methane gas.
4. Conclusion
In this study, antibiotic fermentation broth was treated anaero-
bically, and the reactor performance was investigated with the
modified Stover-Kincannon, first-order, substrate mass balance
and Van der Meer and Heertjes models. During the study, the high-
est COD removal efficiency and methane production were obtained
as 95.7% and 0:30 Lmethane g1
CODremoved , respectively.
According to the results, antibiotic fermentation broth could be
treated effectively using anaerobic process. Because only a limited
number of studies on this subject have been performed, both the
impact of different operating conditions in anaerobic treatment
and the effectiveness of other treatment methods should also be
examined.
Acknowledgement
We thank Hidromer Aritma Sistemleri Ltd. Sti. and Deva Kar-
tepe API Production Plant for supporting the study.
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