2010 International Conference on Challenges in Environmental Science and Computer Engineering

Effect of Substrate Concentration on Methane Fermentation of Cattle Dung

Jha Ajay Kumar, He Junguo, Li Jianzheng and Zheng Guocheng

State Key Laboratory of Urban Water Source and Environment

Harbin Institute of Technology
Harbin 150090, China
Emails: [email protected]; [email protected], [email protected]; [email protected]

Abstract- Methane fermentation of cattle manure (CM) action of various groups of anaerobic bacteria such as
methanogenic bacteria, acetogenic bacteria,
with different total solid contents were conducted in the
fermentative bacteria and so on. During the anaerobic
laboratory-scale batch reactors (2.5 L volume) in order digestion process, complex organic macromolecules
to determine the substrate concentration performance are first hydrolyzed and fermented into simpler
soluble molecules which are then converted by acid
and potential energy recovery at the optimal mesophilic
forming bacteria into volatile fatty acids (such as
(35 oC) and thermophilic (55 oC) temperatures. The acetic acid, propionic acid, butyric acid), carbon
three reactors containing CM and water in the ratios dioxide and hydrogen. Methane is formed by
methanogenic bacteria, either by breaking down the
(w/w) of 1:1 (wet fermentation), 2:1 and 1:0 (undiluted
acids to methane (approximately 70%) or by reducing
manure, dry fermentation) were utilized for the both carbon dioxide with hydrogen (approx. 30%) [2, 3].
conditions. The work was performed for a period of 63 As known, wet anaerobic fermentation of manure
and other organic wastes for methane production has
days to compare the volatile solids and dissolved been widely used and well developed by now [4]. In
organic carbon degradation, COD removal, and VFAs. agriculture-based developing countries, some farmers
The results showed that the specific biogas yields, in have benefited from manure to biogas household
technology. For instance, in Nepal, 189122 domestic
terms of per kilogram of CM, were 41.68, 43.72 and biogas plants have been constructed till now [5].
44.98 L/kg with 22.61, 23.69 and 24.96 L/kg methane Though wet anaerobic digestion is the familiar
contents respectively for the ratios of 1:1, 2:1 and 1:0 at technology for energy production, there are some
remarkable problems with the process, such as
35 oC, while 43.03, 44.89 and 45.71 L/kg biogas with having large digester size, requirement of liquid
23.45, 24.15 and 25.57 L/kg methane contents source, and slurry handling problem etc. Some
respectively for the ratios 1:1, 2:1 and 1:0 at 55 oC. The
factors like less land and no source of water have also
restricted the promotion. On the contrary, the dry
volume of the digester was increased in the wet anaerobic fermentation process stabilizes the manure
fermentation for the same loading rate. Though water and other organic wastes in its produced form. Dry
fermentation is an innovative anaerobic treatment
and thermophilic condition could promote the startup
approach to treat high solid contents bio-wastes
process and the biodegradability of the substrate, the without dilution. It offers great advantages like
total biogas and methane yields in the dry fermentation utilization of wastes in its produced form, no
requirement of liquid source, high organic loading
were found comparable to the conventional process (wet
rate, smaller digester, no liquid effluent, and no
fermentation). requirement of purification of effluent etc [6, 8]. It
Keywords- Cattle manure; Substrate concentration; Dry would be more feasible for semiarid climates and
fermentation; Wet fermentation; Biogas places where no easy access of water [7]. The process
has attracted an increasing attention all around the
world with a view to reduce cost of digesters as well
I. INTRODUCTION
as slurry handling problems [8].
Anaerobic digestion is the process of controlled Because the biodegradation of undiluted as well as
decomposition of organic matter by microbial diluted manures do not vary considerably, the biogas
consortia in an oxygen-free environment to produce production with their methane contents are similar [8].
clean energy and carbon dioxide and reclaim nutrient Dry fermentation of energy crops offers great
rich fertilizer [1]. This process is carried out by co- potential for production of methane and produces

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DOI 10.1109/CESCE.2010.245
 much more bio-energy compared to bio-ethanol and the analysis of various parameters like pH, total COD,
bio-diesel [9]. As dry anaerobic digestion has more soluble COD, TS, VS, VFA, dissolved organic
benefits compared to other technologies, research on carbon, TKN, ammonia N etc. while pH meter was
dry fermentation has great importance to make a set up at the other port.
more efficient and feasible process to solve
multifaceted and dreadful waste problem. In this II.B. Characteristics of feed stocks
study, the wet and dry fermentation are compared The study was conducted to evaluate the mesophilic
taking various substrate concentration i.e. total solids and thermophilic anaerobic digestion of cattle
on the optimum temperatures for the mesophilic and manure at three different initial concentrations of
thermophilic range. total solids in the process. The cattle manure was
obtained from a livestock farm of Harbin, China and
II. METHODS AND MATERIALS was stored at -20 oC. In the fermentation process, the
II.A. Experimental set up and procedure substrate was pretreated and fed into air tight digester
The experiments were carried out in six batch lab- under specified environmental conditions (e.g.
reactors of 2.5 L effective volume with an internal Temperature, pH) for 63 days with or without
diameter of 13 cm, and height of 25 cm. The three dilution. Pretreatment means separation of manure
reactors were kept in a water bath of operational from foreign materials like stones, metals etc. and the
temperature 35oC, the optimum temperature for addition of inoculants (20% digested manure) into the
mesophilic range, while other three were maintained feedstock. Table 1 shows the composition of the
at 55oC, the optimum temperature for the influent for each reactor and physical-chemical
thermophilic range, in an another water bath [6, 10]. characteristics of the initial substrate. For reactors
The cover of each reactor was incorporated with the R1-35-M and R2-55-M, the manure was used in its
two ports: one for measuring the biogas produced and produced form while for R3-35-M &W and R4-55-
to take the sample for the analysis of biogas M&W, the manure was diluted with half amount of
composition while another for adding NaOH to water on weight basis and for the reactor R5-35-
maintain pH. The other two ports were kept on the M&W and R6-55-M&W, the manure was diluted
side of the reactor : one was kept above 5 cm from with equal amount of water on weight basis.
the bottom from which sample could be taken out for

Table 1: Composition and condition of six reactors utilized for the experiments

Density Temperature VS
Reactors Denomination Composition pH TS (%) TS (g/kg)
(g/cm3) (oC) (% TS)
1000g
1 R1-35-M 1.153 35 7.5 15.18 151.8 80.7
manure
1000g
2 R2-55-M 1.153 55 7.5 15.18 151.8 80.7
manure
1000g
3 R3-35-M&W manure + 1.036 35 7.6 10.09 100.9 81.0
500 g water
1000g
4 R4-55-M&W manure + 1.036 55 7.6 10.09 100.9 81.0
500 g water
1000g
5 R5-35-M&W manure + 1.018 35 7.7 7.68 76.8 81.0
1000g water
1000g
6 R6-55-M&W manure+ 1.018 55 7.7 7.68 76.8 81.0
1000g water

II.C. Analytical methods analytical determinations were performed according to

The parameters analyzed were temperature, pH, the standard methods [11]. The yielded biogas was
density, total solids, volatile solids, total chemical measured per day by water displacement method using
oxygen demand (COD) and soluble COD, VFAs, jar of 1 or 2 litres capacity for each reactor. The
total nitrogen Kjeldahl (TNK), ammonia nitrogen constituents (CH4, CO2 and H2) of the biogas were
(NH4-N), dissolved organic carbon (DOC). All the determined using Gas Chromatography (SC-7,

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Shandong Lunan Instrument Factory) equipped with 3 50
a thermal conductivity detector and a 2 m stainless
column packed with Porapak TDS201 (60-80 mesh). 2.5 1 2 3 4
40
Nitrogen was employed as the carrier gas at a flow

B i o g a s y ie ld ( L )
5 6 c1 c2

C u m u l a ti v e b i o g a s ( L )
rate of 40 mL/min. The operation temperatures for 2
the injection port, oven and detector all were 80 °C. c3 c4 c5 c6 30
One gram sample was mixed with 500 mL water 1.5
properly. The mixture was utilized to determine COD. 20
The mixture was filtered to find soluble COD, TKN, 1
NH3-N and DOC. The mixture was centrifuged at
6000 rpm for 15 min. to determine VFAs using Gas 0.5 10
chromatography (GC-122, Shanghai Anal. Inst. Co.)
with a hydrogen flame ionization detector and a 2 m 0 0
stainless column packed with Porapak GDX103 (60-
80 mesh. The operating temperatures for the injection 0 5 10 15 20 25 30 35 40 45 50 55 60 65
port, oven and detector were 190 °C, 220 °C and
220 °C, respectively. Nitrogen was applied as the
Time (d)
carrier gas at a flow rate of 30 mL/min. Fig.1.Biogas yield and cumulative biogas

III. RESULTS AND DISCUSSION

Six lab-batch reactors were tested during a period
of 63 days to evaluate the influence of substrate
concentration on methane fermentation of cattle dung
at three different initial concentrations of total solids 1：R1-35-M；2：R2-55-M；3：R3-35-M&W
in the optimal mesophilic and thermophilic anaerobic 4：R4-55-M&W；5：R5-35-M&W；6：R6-55-M&W
digestion temperatures. The pH of cattle manure was C1：Cumulative R1-35-M; C2:cumulative R2-55-M; C3:
initially around 7.5. It was decreased and maintained Cumulative R3-35-M&W; C4: Cumulative R4-55-M&W; C5:
Cumulative R5-35-M&W; C6: Cumulative R6-55-M&W
stable in between 6.8 to 7.6 through out the
experiment by adding NaOH. The TKN and NH3-N
were less than 2060 and 920 mg/kg during the
fermentation period in all the reactors. So, nitrogen
accumulation did not reach inhibitory levels as the
critical ammonia concentration is 2800 mg/kg NH4-N
[12]. The maintained environmental condition and found that there was higher biogas and methane yield
obtained results were indicative of strong microbial in the reactor R6-55-M&W and R5-35-M&W but the
activities. size of these digesters had to be the double of the
reactors R1-35-M, R2-55-M. The production of biogas
III.A. Biogas generation and methane content and methane at optimal thermophilic temperature (55oC)
Fig.1 shows the specific biogas generation per day was found more than optimal mesophilic temperature
and cumulative biogas yield in litres in all the six (35oC).
reactors. The cumulative specific biogas generation
of the reactors R1-35-M, R2-55-M, R3-35-M&W, III.B. VS, COD and OC removal
R4-55-M&W, R5-35-M&W and R6-55-M&W were Table 2 shows the removal percentage of volatile solids
measured 41.68, 43.03, 43.72, 44.89, 44.98 and 45.11 (VS), total chemical oxygen demand (TCOD), soluble
L/kg with 22.60, 23.45, 23.69, 24.15, 24.96 and chemical oxygen demand (SCOD) and dissolved
25.57 L/kg methane contents, respectively. The organic carbon (DOC) and methane yield per g VS and
cumulative biogas and methane generation showed g COD in biomethanization processes of cattle manure
an increase day after day until reached the maximum with different initial total solids (TS) concentrations at
value and then decreased slowly day after day in the the optimal temperatures: 35 oC for mesophilic range
all reactors. The initial methane contents in the and 55 oC for thermophilic range. It can be observed
yielded biogas has increased and exceeded 50% after that the highest efficiency of VS, COD and DOC
one week in all the reactors. The hydrogen content in removals and methane yield were found in R6-55-
the biogas was observed around 1% initially but it M&W followed by R5-35-M&W, R4-55-M&W, R3-
became negligible after one week. It was detected 35-M&W, R2-55-M and R1-35-M but there were no
that the addition of water and thermophilic condition much differences.
has prompted the start up period with early
generation of biogas and biodegradability. It was also

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Table 2: Organic matter degradation and methane yield in each reactor

Organic matter & its removal Methane yield

Initial Total Initial DOC
Reactor Initial VS Soluble Initial L L
Total COD Soluble degra
VS removal COD DOC CH4/gV CH4/gCO
COD removal COD dation
(g/kg) (%) (%) (g/kg) S D
(g/kg) (%) (g/kg) (%)
1 122.32 47.84 112.78 43.78 58.34 55.82 34.84 30.58 0.3419 0.4619
2 122.32 48.29 112.78 44.42 58.34 57.15 34.84 32.10 0.3584 0.4681
3 81.81 48.84 76.44 44.64 40.44 57.90 23.92 32.88 0.3611 0.4691
4 81.81 49.43 76.44 45.46 40.44 58.98 23.92 34.18 0.3774 0.4695
5 62.17 50.11 58.55 46.13 32.12 61.32 18.14 35.84 0.3790 0.4702
6 62.17 51.13 58.55 47.20 32.12 63.50 18.14 37.32 0.3846 0.4712

III.C. Volatile fatty acids generation 1. Pain, B.F., Hepherd, R.Q., Anaerobic digestion of livestock
In this study, all the reactors showed high fatty acids in wastes. In: Pain, B.F., Hepherd, R.Q. (Eds.), Anaerobic
the start up phase. The degradation of complex organic Digestion of Farm Waste. NIRD Technical Bulletins,
matter into the intermediate VFAs was greater in this Reading, pp. 9-14, 1985.
hydrolysis and acidogenic stages. As methanogenesis 2. McInerney, M. J. and Bryant, M. P., In Fuel Gas
Production from Biomass (ed. Wise, D. L.), Chemical
and percentage methane content in the biogas have
Rubber Co. Press Inc., West Palm Beach, Florida, pp. 26–
stabilized since second week, the VFA concentrations 40, 1981.
decreased with superior acetic acid removal rate. 3. Chynoweth, D. P., Owens J. M., Legrand, R., Renewable
Methane from Anaerobic Digestion of Biomass.
IV. CONCLUSION Renewable Energy 22 (1-3), 1-8, 2000.
The initial substrate concentration persuades the 4. Raphael M. Jingura , Rutendo Matengaifa, Optimization of
anaerobic digestion of the cattle manure. The specific biogas production by anaerobic digestion for sustainable
methane generation of the cattle manure was found to energy development in Zimbabwe, Renewable and
Sustainable Energy Reviews 13 (2009) 1116–1120, 2007.
be 0.3419, 0.3584, 0.3611, 0.3774, 0.3790 and 0.3846
5. BSP Nepal, 2008. http://www.bspnepal.org.np/ (December
m3/kg VS while in terms of m3/kgCOD were 0.4619, 12, 2009).
0.4681, 0.4691, 0.4695, 0.4702 and 0.4712, 6. Ward, Alastair J., Hobbs, Phil J., Holliman, Peter J., Jones,
respectively. The biogas generation and biodegradation David L., Optimisation of the Anaerobic Digestion of
of the substrate have started early in the case of diluted Agricultural Resources, Bioresource Technology, 99(2008)
manure at optimal thermophilic condition followed by 7928-7940.
diluted manure at mesophilic condition than undiluted 7. Kottner, M., Biogas and Fertilizer Production from Solid
manure. There were no significant variations in the total Waste and Biomass through Dry Fermentation in Batch
Method, International Biogas and Bioenergy Centre of
biogas and methane yields and the quality of the biogas
Competence, Germany.
(percentage content of methane) but wet fermentation 8. Bhattacharya, T.K., Mishra, T.N., Biodegradability of
reactor requires larger reactor volume for the same Dairy Cattle Manure under Dry Anaerobic Fermentation
loading rate. Thus, the dry anaerobic fermentation Process, IE(I) Journal-AG vol 84, University of
might be a promising alternative to the conventional Agriculture and Technology, Uttaranchal, India, 2003.
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10. Tchobanoglous G., Burton F.L., Waste-water
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ACKNOWLEDGEMENTS
York: McGrraw-Hill; p1334, 1991.
The authors would like to thank the National High
11. APHA, Standard Methods for the Examination of Water
Technology Research and Development Program of China
and Wastewater [J]. 19th ed., American Public Health
(863 Program, Grant No. 2006AA05Z109), National S&T
Association; Washington, DC, 1995.
Major Projects (Grant No. 2008ZX07207-005-02) and Harbin
Science and Technology Bureau (Grant No. 2009RFXXS004) 12. Poggi-Varaldo, H. M., Rodriguez-Vazquez, R.,
for their supports for this study. Fernandez-Villagomez, G., Esparza-Garcia, F., Inhibition
of mesophilic solid-substrate anaerobic digestion by
ammonia nitrogen. Appl. Microbial, Biotechnol. 47, 284-
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