Thermophilic Anaerobic Treatment of Cassava

Ethanol Wastewater in Continuously Stirred Tank

Reactor (CSTR)
Li Xie, Gang Luo, Qi Zhou, Wenjing Sang, Jiawei Sun
College of Environmental Science and Engineering
Tongji University
Shang Hai, China
sally.xieli@gmail.com
AbstractThe high content of organic matter and suspended
solids with relatively low pH of 4 characterizes the cassava
ethanol wastewater. A lab-scale anaerobic continuously stirred
tank reactor (CSTR) was used under thermophilic condition
(55
o
C) to treat cassava ethanol wastewater. Experimental results
showed that successful start up of the reactor inoculated with
mesophilic granular sludge could be obtained after 40 days
continuous operation with step-wise increases in the organic
loading rate. The organic loading rate could reach as high as
14kgCOD/(m
3
d) by decreasing the hydraulic retention time to 5
days. The performance of reactor was stable with TCOD removal
efficiency of 90% and biogas production of 18L/d containing
about 60% methane. Moreover, significant fluctuation in
performance of reactor was not observed with the changes of
organic loading rate. And results showed adjustment of pH or
alkalinity is not necessary due to the sound buffer capacity of the
system.
Keywords-thermophilic anaerobic; CSTR; cassava ethanol
wastewater; suspended solids
I. INTRODUCTION
Utilization of cassava for ethanol production could provide
the most effective use of natural resources, but generate around
15 tons wastewater with high strength organics for each ton
cassava ethanol production during the fermentation and
distillation process. The high content of organic matter and
solid residues from cassava fragments and peels are of great
environmental concern if not properly treated. Such wastewater
is well suited to anaerobic treatment and there are many high
efficiency reactors to treat cassava wastewater such as UASB.
However high suspended solids (SS) content may decrease the
capacity of UASB, it is common practice to remove solids
before treatment, therefore only the soluble part of the
wastewater with small amount of residual solids (up to 500
mg/L) is admitted to use the UASB reactor [1]. The SS
concentration of cassava ethanol production wastewater could
be as high as 30000mg/L, and the solids could not be easily
separated due to the high viscosity of wastewater and therefore
lower the performance of UASB. Additionally, the separated
solids contain low content of protein, fat and high
concentration of cellulose, which limits its usage as feed for
animals [2]. Therefore, it was necessary to develop an
anaerobic technology which could endure high solids
concentration for the treatment of cassava ethanol wastewater
without decantation process.
Anaerobic continuously stirred tank reactor (CSTR) is
widely used for high solids content organic material of
wastewater, such as sludge, piggery wastewater, food wastes
[3] [4], and herein the solids could be partly converted to
biogas without any separation. Meanwhile, CSTR could
provide improvement in performance and stability of wastes
treatment. In recent years, thermophilic anaerobic process has
gained much attention primarily due to its higher loading
capacity. Moreover, Mladenovska and Ahring observed that
specific biogas production rates were higher under
thermophilic conditions than that of under mesophilic
conditions, attributed mainly to the higher maximum specific
growth rate (2-3 times) of thermophilic microbes than that of
mesophilic analogues [5]. This higher gas production may lead
to an improvement in the energy balance of the biogas
technology, and thereby incur less capital cost as a result of
smaller digester size [6].In thermophilic condition, start-up of
reactor is crucial to establish a proper microbial community.
Poor start-up in biological treatment systems could lead to
prolonged period of acclimation [7] and ineffective removal of
organic matter [8].
Considering the temperature of cassava ethanol wastewater
above 60
o
C, the application of thermophilic anaerobic CSTR in
such wastewater may obtain the advantage of thermophilic
anaerobic. Meanwhile, it is not necessary to remove solids
before CSTR and SS can partly convert to biogas, which can
solve the problem of difficult disposal of separated solids.
Therefore the objectives of this study were to evaluate the
feasibility of thermophilic anaerobic CSTR for cassava ethanol
wastewater treatment and operational conditions such as pH,
alkaline, and biogas production in the reactor were investigated.
II. MATERIALS ANDMETHODS
A. Cassava ethanol wastewater
Cassava ethanol wastewater used in this study was
collected directly from Taicang cassava ethanol plant, located
in Jiangsu province, China. After collected, samples were
pretreated and submitted for analysis.
B. Experimental Set-up and Start-up Conditions
Laboratory-scale equipment was illustrated in Fig.1. An
automatic biological fermentation tank (5L) (Shiyuan,
Shanghai) was used as CSTR with working volume 4L. The
This work was sponsored by the Bayer Sustainable Development Foundation
and the Science Committee Project of Shanghai (No. 062307038)
978-1-4244-1748-3/08/$25.00 2008 IEEE 3001
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Fig.1.Schematic Diagram of Experimental Equipment
headspace about 1L was connected to a biogas collector. The
temperature was automatically maintained at 551
o
C in a
heated water bath. The pH of the mixture could be measured
on-line by the installed probe. Wastewater was pumped into
the feeding port, and was continuously mixed at 200rpm by a
mechanical stirrer in the reactor. The effluent was pumped
from the outlet port and separated in the followed settling tank.
Settled sludge was recycled to reactor periodically with
recycling ration of 1:1.
CSTR was started with granular sludge as inoculum from a
mesophilic UASB reactor of Taicang cassava ethanol plant
(Jiangsu Province, China). The pH, volatile suspended solids
and total suspended solids concentration of the inoculums were
7.5, 42g/L, 70g/L, respectively. The reactor was fed with 1L of
granular sludge and 3L raw wastewater diluted with tap water
by 5 times initially, and then operated with stirring for 3 days
without influent and effluent until the pH raised from 6.50 to
7.10. After that the CSTR was continuously operated for 2
weeks until the TCOD removal efficiency was 80%, and then
the organic loading rate (OLR) of the reactor was increased
step by step by decreasing the dilution times of raw wastewater.
Raw wastewater could be directly fed to the reactor after 53
days operation with stable TCOD removal efficiency of 90%.
Successful start-up of CSTR was obtained and later the
hydraulic retention time (HRT) was gradually shortened to
increase the OLR.
C. Analytical Methods
Influent and effluent COD, pH, Alkalinity, VFA, biogas
production were daily determined during the reactor operation.
The other parameters, SS and methane, were determined twice
every week.
Total and soluble COD, SS, VSS, bicarbonate alkalinity
were analyzed in duplicate according to Standard Methods of
APHA [9]. Organic and volatile fatty acids (VFA) were
determined in triplicate by gas chromatograph (HP5890II-FID)
using a CPWAX52CB column. Biogas flow was measured
with a device based on liquid displacement by the outgoing
biogas, and methane content of biogas was determined by
using a Shimadzu GC-8A gas chromatograph equipped with a
flame ionization detector (FID) and a 10m Heliflex AT-1000
capillary column.
Samples were centrifuged at 3500 rpm for 10 min and
filtrated through 0.45m filters for the determination of SCOD,
VFA.
III. RESULTS AND DISCUSSION
A. Characteristics of Cassava Ethanol Wastewater
TABLE I. CHARACTERISTICS OF CASSAVA ETHANOL WASTEWATER
Table 1 shows the physicochemical characteristics of the
cassava ethanol wastewater. The temperature of the wastewater
was about 60
o
C. High concentrations of COD and SS in the
range of 40000-70000mg/L and 20000-30000mg/L
respectively characterized the wastewater with relatively low
pH of 4.0. Soluble fraction of the COD varied between 50%
and 75%. The ratio of COD: N: P of the wastewater was about
200:5:1, which was suitable for its bioconversion to methane,
and it is not necessary to add nutrients into the anaerobic
reactor.
B. Performance of CSTR
Fig.2 shows the effect of varying OLR on TCOD removal
efficiency of the CSTR during the start-up and operation period.
OLR was increased progressively at constant step of
1.0kgCOD/(m
3
d) during the start-up period. Initially, the
TCOD of the diluted waste water was about 8000mg/L and the
average TCOD removal efficiency was 60%, which was
contributable to the inadaptability of mesophilic sludge to
thermophilic environment. After 20 days operation
continuously, the inoculum was gradually acclimated to the
Fig.2. Variation of OLR and TCOD removal efficiency during start-up and
operation of CSTR
Parameter Value
Total COD(mg/L) 4000070000
Soluble COD(mg/L) 3000035000
SS(mg/L) 2000030000
pH 4.04.2
Total N(mg/L) 800900
Total P(mg/L) 200400
automatic control
device
CSTR pump
distributing tank
settling tank
effluent
temperature probe
pH probe
biogas
mechanical
mixer
water
bath
gas gathering equipment
3002
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VFA
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Alkalinit y
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methane yield(TCOD)
methane yield(SCOD)
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Biogas Production
Fig.3. Variation of acidity and alkalinity during start-up and operation of
CSTR.a.Variation of Alkalinity and pH in the reactor; b.relation of VFA to
OLR
thermophilic condition, and 90% of the TCOD removal
efficiency was obtained. The reactor was gradually adapted to
the raw wastewater at the corresponding OLR of
5.0kgCOD/(m
3
d) with stable TCOD removal efficiency of
90% (Fig.2). After the start-up period (days 1-55), OLR was
continuously increased from 5.0kgCOD/(m
3
d) to
14.0kgCOD/(m
3
d) at the step of 2-3kgCOD/(m
3
d) with the
decrease of HRT from 10 days to 5 days, and TCOD removal
efficiency could still maintain at 90%. Sun and his colleagues
observed that the changes of OLR below 7kgCOD/(m
3
d) could
lead to obvious fluctuation in COD removal efficiency in the
study of stability of anaerobic baffled reactor [10]. However, in
this study, stable operation performance of thermophilic
anaerobic CSTR was observed with the increase of OLR, even
at 14kgCOD/(m
3
d). The performance of reactor was not
affected by the SS concentration of the wastewater. The
effluent SS concentration was lower than 3000mg/L with stable
removal efficiency of above 80% in the whole operation period.
The changes of operational parameters of pH and alkalinity
at different OLR are presented in Fig.3a. The pH of the mixture
was stabilized around 7.30 in the whole period with the
influent pH of 4.2. The alkalinity index could further reflect the
buffer capacity of reactor. As shown in Fig.3a, the alkalinity in
start-up period and operation period is between 1500~2000mg
CaCO
3
/L and 2000~5000mg CaCO
3
/L, respectively. The
higher OLR of 10-14kgCOD/(m
3
d) didnt lead to the decrease
of alkalinity in reactor, which may contributed to the maturity
Fig.4. Variation of Biogas in the start-up and operation process of CSTR.
a.Relation of biogas production to OLR; b.Variation of Methane yield in the
reactor
of anaerobic sludge obtained from start-up period. The range of
alkalinity obtained in this study is well in agreement with
Hashimotos research that alkalinity levels in anaerobic
digesters should maintain from 2500 to 5000mg CaCO
3
/L for
methane production [11]. Additionally, Fig.3b shows that the
total VFA concentration in reactor was almost below 200mg/L
at different OLR. The results described above indicated that the
thermophilic CSTR has strong buffer capacity for cassava
ethanol wastewater and could operate stably without influent
pH adjustment or alkaline addition into the reactor.
C. Biogas Production
The volumetric biogas production (L biogas/d) increased
with the increase in ORL as Fig.4a presents. Biogas production
of 18L/d was obtained at the highest OLR of 14kgCOD/(m
3
d).
Jin et al. noticed that OLR above 4kgCOD/(m
3
d) could inhibit
biogas production when using mesophilic anaerobic digester to
treat food waste [12]. However, no inhibition to biogas
production was observed in this study even at OLR of
11~14kgCOD/(m
3
d). Previous study of temperature effect on
reactor performance has confirmed that organic loading
potentials of anaerobic reactors in thermophilic condition are
substantially higher than that in mesophilic condition and the
reaction rate proceed faster [13]. Methane content in the biogas
was in the range of 55% to 61%. Specific methane yield
calculated by TCOD and SCOD respectively in the whole
period is shown in Fig.4b. The methane yield calculated by
TCOD was relatively stable in the range of 0.20~0.25m
3
, and
the value was obviously lower than theoretical methane yield
3003
of 0.35m
3
/COD removal. However, the methane yield
calculated by SCOD was from 0.37 m
3
to 0.52 m
3
that was
higher than theoretical value. The relatively lower or higher
methane yield counted by TCOD and SCOD should be
attributable to the high SS content of wastewater. Most of SS
was converted to SCOD in the anaerobic reactor, which led to
the higher methane yield calculated by SCOD. More studies
should be conducted to understand the transformation of SS in
CSTR in near future.
IV. CONCLUSIONS
The high content of organic matter and SS from cassava
fragments and peels with relatively low pH of 4 characterizes
the cassava ethanol wastewater. Thermophilic anaerobic
treatment of high strength cassava ethanol wastewater in CSTR
is a robust and reliable process. The overall conclusion from
the research results can be summarized as:
Successful start-up of thermophilic anaerobic CSTR
inoculated with mesophilic granular sludge could be
achieved with the increase in OLR gradually (step by
step) at 1kgCOD/(m
3
d). Raw wastewater without any
dilution could be fed into reactor and the OLR was
5kgCOD/(m
3
d).
In the operation period, the OLR was increased at the
step of 2~5kgCOD/(m
3
d), with the decrease in HRT.
After 80-days operation, the OLR could be as high as
14kgCOD/(m
3
d) with stable TCOD removal
efficiency of 90%, biogas production of 18L/d
containing about 60% methane.
The pH of the mixture was stabilized around 7.30 and
the range of alkalinity was from 2000 to 5000 mg
CaCO
3
/L, indicating the system has good buffer
capacity and influent pH adjustment or addition of
alkaline into reactor is not required.
The methane yield counted on SCOD was
0.37~0.52m
3
, which is higher than theoretical value
due to the bioconversion of SS to SCOD.
ACKNOWLEDGMENT
The authors wish to thank Taicang cassava ethanol plant for
their raw cassava ethanol wastewater samples and valuable
practical experience.
REFERENCES
[1] G.D. Zupani, M.Straiar, M.Ro, Treatment of brewery slurry in
thermophilic anaerobic sequencing batch reactor, bioresource
technology, vol 98, pp 2714-2722, 2007.
[2] K.X.Li, T.P.Wang, Z.J.Zhang, Study on the technology of potato
ethanol wastewater treatment, Chinese Journal of Industrial Water
Treatment, vol 25, pp 13-15, 2005.
[3] I.Angelidaki, X.Chen, J.Cui, P.Kaparaju, L.Ellegaard, Thermophilic
anaerobic digestion of source-sorted organic fraction of household
municipal solid waste: Start-up procedure for continuously stirred tank
reactor. Water Reasearch, vol 40, pp 2621-2628, 2006.
[4] M.A.Rubia, M.Perez, L.I.Romero, D.Sales, Effect of solids retention
time on pilot scale anaerobic thermophilic sludge digestion, Process
Biochemistry, vol 41, pp 79-86, 2005.
[5] Z.Mladenovska, B.K.Ahring, Growth kinetics of thermophilic
Methanosarcina spp. Isolated from full-scale biogas plants treating
animal manure. FEMS Microbiol. Ecol, vol 31, pp225-229, 2000.
[6] H.Bouallagui, O.Haouari, Y.Touhami, R.BenCheikh, L.Marounani,
M.Hamdi, Effect of temperature on the performance of an anaerobic
tubular reactor treating fruit and vegetable waste, Process Biochem, vol
39, pp2143-2148, 2004.
[7] J.H.Wu, W.T.Liu, I.C.Tseng, S.S.Cheng, Characterization of a 4-
methylbenzoate-degrading methanogenic consortium as determined by
small-subunit rDNA sequence analysis. Journal of Bioscience and
Bioengineering, vol 91, pp 449-455, 2001.
[8] M.E.Griffin, K.D.McMahon,R.I.Mackie, L.Raskin, Methanogenic
population dynamics during start-up of anaerobic digesters treating
municipal solid waste and biosolids. Biotechnol. Bioeng, vol 57, pp
342355, 1998.
[9] APHA(American Public Health Association, American Water Works
Association, Water Pollution Control Federation). Standard Methods for
the examination of water and wastewater.17
th
ed. Washington: Rhodes
Trussell Eds, 1995.
[10] J.H.Sun, J.F.Wu, B.Zhang, Effect of shock loads on performance of an
anaerobic reactor, Chinese Journal of Technology of Water Treatment,
vol 30, pp 362-364, 2004.
[11] A.Hashimoto, Y.Chen, V.Varel, S.Robinson, Anaerobic fermentation
of agricultural residue.In:M.L.Shuler,Editor,Utilization and Recycle of
Agricultural Wastes and Residues, CRC Press, Bocar Raton, Florida, pp
145-146,1980.
[12] J.X.Jin, S.H.Wang, XF Xiao, C.S.Yang, Experimental Study on
Traditional Anaerobic Digestion Process for Food Waste Treatment,
Chinese Journal of China Water & Wastewater, vol 23, pp 55-57, 2007.
[13] J.H.Ahn, C.F.Forster, The effect of temperature variations on the
performance of mesophilic and thermophilic anaerobic filters treating a
simulated papermill wastewater, Proc Biochem, vol 37, pp 589-
594,2002.
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