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ScienceDirect
Energy Procedia 52 (2014) 328 336

2013 Alternative Energy in Developing Countries and Emerging Economies

Anaerobic Co-digestion of Canned Seafood Wastewater with

Glycerol Waste for Enhanced Biogas Production
Kiattisak Panponga,, Galaya Srisuwana, Sompong O-Thonga,b,c*, Prawit Kongjand
b

a
School of Engineering and Resources, Walailak University, Nakornsrithamarat 80161, Thailand
Department of Biology, c Research Center in Energy and Environment, Faculty of Science, Thaksin University, Phatthalung 93110,
Thailand
d
Department of Science, Faculty of Science and Technology, Prince of Songkla University, Pattani 94000, Thailand

Abstract
The potentiality in biogas production from anaerobic co-digestion of canned seafood wastewater (CSW) with
glycerol waste (GW) was investigated. Methane yields from anaerobic co-digestion of CSW with 1, 2, 3, 4, 5, 6, 7, 8,
9 and 10% (v/v) of GW were 577, 265, 101, 51, 11, 9, 4, 3, 2 and 2 mLCH4/g VS-added, respectively. The anaerobic
co-digestion of 99% CSW with 1%GW was the optimal mixture ratio for methane production with a maximum
methane yield was 577 mLCH4/g VS-added and 97% biodegradability. Meanwhile, the maximum methane yield of
1%GW and 100%CSW were 211 and 278 mLCH4/g VS-added. Methane yield increased by 108% when compared
with digested CSW alone. The maximum methane production from anaerobic co-digestion of 99%CSW with 1%GW
was 5.8 m3 CH4/m3 of mixed wastewater and electricity production of 1 m3 mixed wastewater would be 207 MJ or 58
kWh of electricity. Continuous methane production from anaerobic co-digestion of CSW with 1% GW in up-flow
anaerobic sludge blanket (UASB) reactors gave methane production rate of 2.33 LCH4/L-reactor.day.

Elsevier
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under and/or
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2014
2013
Published
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Selection and peer-review under responsibility of the Organizing Committee of 2013 AEDCEE

of the Research

Keyword: Co-digestion, Biogas production, Canned seafood wastewater, Glycerol waste, Synergistic effect ;

1. Introduction
Canned seafood industry (tuna, sardine, mackerel, etc.) is one of the major exports of Thailand. Most
of the seafood processing plants are located in the southern and eastern coast of Thailand. The amount of
wastewater from canned seafood industry ranges from 300 to 500 m3/day [1]. Canned seafood industry

* Corresponding author. Tel.: +66 746 93992 ; fax: +66 746 93992.
E-mail address: sompong.o@gmail.com

1876-6102 2014 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Selection and peer-review under responsibility of the Organizing Committee of 2013 AEDCEE
doi:10.1016/j.egypro.2014.07.084

Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

discharges the wastewater around 14 to 22 m3/ton of raw material (from washing (9%), cooling (34%),
thawing (26%), ice removal (21%) and sterilization (10%) [1, 2]. Presently, wastewater treatment systems
operating in the canned seafood industry are activated sludge, aerated lagoon, oxidation pond and
anaerobic lagoon, but anaerobic reactors are used less [2]. The wastewater treatment is particularly
difficult because of the high content salts protein and oil [2]. As a result, anaerobic reactor was not
popular due to the problem of high content of organic nitrogen in wastewater which inhibited the
anaerobic process [3]. The inhibitors generally occur in the canned seafood wastewater consisting of salts,
fat, oil, grease and ammonia [2]. Compositions of canned seafood wastewaters were contained of BOD5
(1003,000 mg/L), COD (1,00018,000 mg/L) and nitrogen content (801,000 mg/L) [4]. The total
ammonia nitrogen (TAN) is a combination of free ammonia nitrogen (NH3) and ionized ammonia
nitrogen (NH4+) which will be inhibited methanogenic activity by the concentrations of TAN in the range
of 0.17 to 14 g/L reduced the methane production about 50% [5-7]. Guerrero et al. [8] reported that the
toxicity of free ammonia nitrogen (NH3) for mesophilic conditions was in the range of 25 - 140 mg
NH3/L. Additionally, the anaerobic treatment of canned seafood wastewater is inhibited by the presence
of high sodium or chloride concentrations [2]. Methanogenesis is strongly inhibited by a sodium
concentration of more than 10 g/L [4]. The maximal biogas yield was 0.75 m3/kg COD for anaerobic
filter treatment of fishery wastewater which was operated at OLR of 1.3 kg COD/m3.day and HRT of
11days [3]. Therefore, the investment construction of biogas system in the canned seafood industry is not
worth due to the low biogas production and also poor quality of biogas. One approach to increase the
biogas production in the canned seafood wastewaters is the use the co-digestion process.
Glycerol waste (GW) is a by-product of biodiesel production. By one kilogram of biodiesel
production generated glycerol waste about 100 g or approximate 10% of raw material [9, 10]. Currently,
glycerol waste has a low price because of excessive supplies [9]. Advantages of glycerol are easily
digested, high COD, low prices and stored at room temperature for a long time [11]. Glycerol waste was
used as a co-substrate to improve the biogas production in the anaerobic fermentation process [12].
Glycerol waste could improve C/N ratio and dilute the toxic compounds by the values of C/N and COD/N
ratios were 20 and 70 which suggested a value in the process of anaerobic digestion [13]. Astals et al. [14]
reported that the co-digestion between pig manure and 4% of glycerol waste can be increased the biogas
production of about 400%. The maximum methane yield of 0.32 ml CH4/g COD-removed was achieved
at a mixing ratio of 80:20 (glycerol: pig manure) [15]. Co-digestion of pig manure (PM) with fish waste
(FW) or biodiesel waste (BW) could upgrade biogas volume and composition with compared to sole PM
digestion [16]. The addition of 2 ml of glycerol waste per litter of potato wastewater could be increased
biogas production by 0.74 L biogas/mL glycerol product [11]. Additionally, the glycerol is an
intermediate product of anaerobic degradation of fats. Lipids are hydrolysed to glycerol and free long
chain fatty acids (LCFAs) as the first step in anaerobic conditions [17].
This study aimed to enhance biogas production from canned seafood wastewater (CSW) by codigestion with glycerol waste from biodiesel industry.
2. Methodology
2.1 Granule sludge
The granular sludge samples were collected from wastewater treatment plant of the Kiang Huat Sea
Gull Trading Frozen Food Public Company Limited (KST in Hat-Yai District (Songkhla, Thailand). The
granular size was used in the experiment between 0.8 and 1 mm. The volatile suspended solid (VSS) of
the granular sludge was 33.876 g VSS/L.

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

2.2 Canned seafood wastewater and glycerol waste

The wastewater sample was collected from Kuang Pei San Food Products Public Co., Ltd. in Muang
Trang district, Trang Province, Thailand. The sample was stored at a temperature of 40C before it was
analyzed and used in the experiment. The glycerol waste was collected from the biodiesel plant at Prince
of Songkla University in Hat-Yai District, Songkhla Province, Thailand. The main characteristics of the
canned seafood wastewater and glycerol waste were shown in Table 1.
Table 1. Characteristics of CSW and GW used in the experiments
Parameter
pH
COD(g/L)
VFA(mg/L)
ALK(mg/L)
TN(mg/L)
TP(mg/L)
TS(g/L)
VS(g/L)
Protein(g/L)
Carbohydrate(g/L)
Fat(g/L)
C/N ratio
Na+(mg/L)
K+(mg/L)

CSW
6.3
10.4
2,230
2,560
870
53.6
9.37
7.76
3.90
1.91
0.13
11
560
-

GW
8.8
1,760
6,650
35,050
1,670
71,500
969
910
1.28
845
63.76
949
20
40

2.3 Batch reactor

Anaerobic co-digestions of canned seafood wastewater (CSW) with glycerol waste (GW) were tested
in 1,000 ml serum bottle with a working volume of 900 ml. The serum bottles were fitted with gas
sampling septa closed with rubber stoppers and sealed with aluminium caps. Initially, the digestion
mixtures were flushed with nitrogen gas for 5 min to replace the air (oxygen) in order to achieve
anaerobic conditions. All batch experiments were conducted under mesophilic condition with 11 different
concentrations of GW in the range of 0-10 % (v/v) to determine the appropriate amount of glycerol waste
for the best methane production. The volume of granular sludge of 125 mL (4.23 g VSS) was added in all
experiments. The mixed ratio of CSW with GW had the concentrations of VS in the range of 9.98 90.96
g VS/L (Table 2). During the experiments, biogas was daily collected by water displacement. The
composition of biogas was analysed periodically by GC-TCD. COD, VS, VFA, Alkalinity, VFA/Alk
ratio, pH and Alk/COD ratio were determined from liquid samples at the end. In the continuous operation,
the optimum condition from the batch experiment was selected to operate in continuous reactor (UASB).
2.4 Continuous reactor
Continuous anaerobic co-digestion of CSW with GW was operated in up-flow anaerobic sludge
blanket (UASB) at working volume 2.58 L. UASB reactor constructed by using clear acrylic in thickness
about 0.5 mm. The experiment operated in the optimum condition of batch test (99%CSW+ 1%GW) and
organic loading rate (OLR) of 2 g COD/L. day. The start-up of UASB system was started at OLR of 0.5 g
COD/L. day then gradually increased to 2 g COD/L. day by adjusting the flow rate of feeding. The HRT
was gradually reduced from 51 to 13 days. The biogas production was collected by water displacement
(gas counter) every day. The biogas samples were taken from the UASB reactor everyday for biogas
composition analysis by GC-TCD. The liquid samples were taken from the UASB reactor everyday to
analyze the pH and VFA.

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

Table 2. Initial conditions used in the experiments

Experiment
CSW
GW(1%)
CSW+ GW(1%)
CSW +GW(2%)
CSW +GW(3%)
CSW +GW(4%)
CSW +GW(5%)
CSW +GW(6%)
CSW +GW(7%)
CSW +GW(8%)
CSW +GW(9%)
CSW +GW(10%)

pH
6.30
8.00
6.90
7.10
7.40
7.60
7.80
7.90
8.00
8.10
8.20
8.30

COD
(g/L)
10.4
16.0
25.6
35.2
64.0
88.0
112
128
144
160
176
192

VS
(g/L)
7.76
4.50
9.98
12.20
24.42
36.64
48.86
51.08
63.30
75.52
83.74
90.96

TN
(g/L)
0.870
0.019
0.887
0.993
1.004
1.027
1.047
1.037
1.049
1.061
1.072
1.083

C/N
ratio
11
758
26
32
57
77
96
111
124
136
148
160

COD/VS
(g COD/g VS)
1.34
3.56
2.57
2.89
2.62
2.40
2.29
2.51
2.27
2.12
2.10
2.11

2.5 Analytical methods

pH was measured by using Sartorius Docu - pH meter. Chemical oxygen demand (COD), total solid
(VS), volatile suspended solid (VSS), total nitrogen (TN), total phosphorus (TP), volatile fatty acid,
alkalinity, protein, carbohydrate and fat were analyzed according to standard method for the examination
of water and wastewater [18]. The methane content was analyzed by a gas chromatography (GC)
equipped with a thermal conductivity detector [19]. The synergism calculated using the methane
production of the optimal mixture ratio of CSW with GW (%) in batch test by comparing with the
methane production of CSW and GW(%) alone [20]. Theoretical methane potential calculated according
Bushwells formula which derived from stoichiometric conversion of the compound to CH4, CO2 and NH3
[21].
3. Result and Discussion
3.1 Substrate characterization
The characteristics of CSW and GW before and after mixing were shown in Table 1 and 2. The CSW
was high protein and had a C/N ratio as 11 which was very low when compared to GW. GW consisted
mainly of carbohydrate and had a C/N ratio very high which had a value as 949. Additionally, adding of
GW into CSW at a concentration of 1-10% (v/v) could be increased the C/N ratio between 26 and 160.
Li et al. [22] reported that the suitable C/N ratio for anaerobic digestion in the range 20 30. As a cosubstrate, GW can be adjusted the C/N ratio and pH of CSW. Particularly, it can be balanced the C/N
ratio of the mixed wastewater and it can be diluted the ammonium nitrogen concentration in the anaerobic
digester [23]. It was found that CSW had a COD concentration of 10.4 g/L which not economic feasibility
for biogas production. However, the COD concentration of GW was very high which had a value as 1,760
g/L. So, using GW co-digested with CSW can be increased the COD concentration.
3.2 Methane potential of co-digestion CSW with GW
The accumulative methane production and methane yield by the co-digestion of the CSW and GW
were shown in Fig 1 and 2. Methane yields of co-digestion CSW with 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10%
(v/v) of GW were 577, 265, 101, 51, 11, 9, 4, 3, 2 and 2 mLCH4/g VS-added, respectively. The codigestion of 99%CSW with 1%GW was the optimal mixture ratio for methane production which had a
cumulative methane production of 5,184 mLCH4 and methane yield of 577 mLCH4/g VS-added with 97%

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

biodegradability (Table 3). The maximum methane yield of 100%CSW was 278 mL CH4/g VS-added
with 95% biodegradability. Meanwhile, the maximum methane yield of 1%GW was 211 mL CH4/g VSadded with 70% biodegradability. Methane yield was increased by 108% when comparing with digested
CSW alone. These results showed that adding 1% GW into CSW can be enhanced biogas production.
Maldenovska et al. [24] reported that the co-digestion between manure and lipids gave a methane yield of
382 mLCH4/g VS-added. Additionally, adding 1%GW into CSW also increased the C/N ratio from 11 to
26 (Table 2). Addition of GW at 2-10% (v/v) into CSW increased C/N ratio between 32 and 160 but had a
negative effect on methane production due to system overload. The methanogenic process was inhibited
when increased glycerol waste concentration up to 2%. Fountoulakis et al. [25] reported that adding GW
not exceeded a limiting 1% (v/v) can be boosted the biogas yield. The initial pH after adding GW into
CSW ranged 6.9 8.3 which had suitable for the methanogenic process more than CSW alone (pH 6.3)
(Table 2). In addition, the advantage in using GW as a co-substrate can be adjusted the pH of CSW which
could save the cost of chemicals in adjusting the pH value. The final volatile fatty acid and alkalinity were
in the range of 1,100 7,100 mg/L and 3,100 4,450 mg/L. As a result, the VFA/Alk ratio had a value in
the range of 0.25 2.29 which indicated the efficiency of anaerobic digestion (Table 3). The suitable
VFA/Alk ratio for anaerobic digestion must be no greater than 0.4 [26].
Table 3. Summary of methane yield, biodegradability and pH of anaerobic co-digestion of CSW with GW.

Exp.

Initial
Loading
(gVS/L)

CH4 Yield
(mL CH4/g VS
added)

biodegradability
(%)

pH After
digestion

VFA/Alk
ratio

7.76
4.50
9.98
12.20
24.42
36.64
48.86
51.08
63.30
75.52
83.74
90.96

278
211
577
265
101
51
11
9
4
3
2
2

95
70
97
80
78
77
70
70
62
56
36
33

6.7
5.1
7.3
6.8
5.3
5.0
4.8
4.8
4.7
4.8
4.9
4.8

0.31
3.26
0.25
0.76
1.71
1.92
1.81
1.69
1.83
1.99
2.12
2.29

CSW
GW(1%)
CSW+ GW(1%)
CSW+GW(2%)
CSW+GW(3%)
CSW+GW(4%)
CSW+GW(5%)
CSW+GW(6%)
CSW+GW(7%)
CSW +GW(8%)
CSW +GW(9%)
CSW+GW(10%)
5500

CSW
CSW+2%GW
CSW+4%GW
CSW+6%GW
CSW+8%GW
CSW+10%GW

Cumulative methane production (mL CH4 )

5000
4500
4000

CSW+1%GW
CSW+3%GW
CSW+5%GW
CSW+7%GW
CSW+9%GW

3500
3000
2500
2000
1500
1000
500
0
0

10

15

20

25

30

35

40

45

Time (days)

Fig. 1. Cumulative methane production from anaerobic co-digestion of CSW with GW.

50

55

60

64

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

700

M eth an e y ield (m L C H4 /g VS-ad d ed

600
500
400
300
200
100

C SW +1 0 % GW

C SW +9 % GW

C SW +8 % GW

C SW +7 % GW

C SW +6 % GW

C SW +5 % GW

C SW +4 % GW

C SW +3 % GW

C SW +2 % GW

C SW +1 % GW

C SW

Fig. 2. Methane production from anaerobic co-digestion of CSW with GW.

3.3 Synergism of co-digestion between CSW and GW

The anaerobic co-digestion of 99%CSW with 1%GW resulted in positive synergism by increasing the
methane production, methane yield and degradation efficiency. The total methane production was 5,184
mLCH4 while total methane production from CSW and GW alone were 1,945 and 951 mLCH4. The
synergistic methane production (Syn-MP) of co-digestion between CSW and GW was 2,288 mLCH4 (Fig
3A). Additionally, the methane yield of co-digestion was 577 mLCH4/g VS-added (Theoretical methane
yield = 630 mLCH4/g VS-added). Methane yield was increased by 108% when compared to the methane
yield of CSW (278 mLCH4/g VS-added) and GW (211 mLCH4/g VS-added). The synergistic methane
yield (Syn-MY) was 88 mLCH4/g VS-added (Fig 3B). The degradation efficiency increased from 95% to
97% after adding 1%GW into CSW. The maximum methane production from anaerobic co-digestion of
CSW with 1% GW was 5.8 m3 CH4/ m3 of mixed wastewater. Meanwhile, the maximum methane
production of CSW and GW alone was 2.2 m3 CH4/m3 of CSW wastewater and 0.2 m3 CH4/m3 of 1%GW
wastewater. Finally, the electricity production of 1 m3 of mixed wastewater would be 207 MJ or 58 kWh
of electricity by calculation from the content of energy as 36 MJ/m3 CH4 and 10 kWh/m3 CH4 which had
a conversion efficiency of approximate 40% in a gas motor [20].
3.4 Continuous methane production of co-digestion CSW with GW in UASB reactor
Anaerobic co-digestion of 99%CSW and 1%GW was selected to operate in the UASB reactor.
Granular sludge in the UASB could adapt quickly because the granular sludge was taken from the UASB
treating the same type of wastewater. The first phase, the wastewater entering into the UASB system at
OLR 0.5 g COD/L. day was acclimatized phase. The OLR was gradually increased up to 2 g COD/L. day
by adjusting the inflow rate of wastewater (fixed initial COD was 25,600 mg/L). The results showed that
the biogas production rate, methane composition and pH were 2.33 L/L reactor-day, 60% and 6.9 -7.2 in
steady state (Fig 5A). Khanal [27] reported that the optimal pH for anaerobic process in the range 6.8
7.4. The methane yield of 309 mLCH4/g COD-removed (640 mL CH4/g VS-added) with 80% COD
removal achieved under steady state (Fig 5B). The result of this experiment was nearby to that of
Nuchdang and Phalakornkule [15] which reported that the maximum methane yield of 320 mL CH4/g
COD-removed at an OLR of 1.6 g COD/L. day in a case of the co-digestion between glycerol and pig
manure. Volatile fatty acids (VFAs) were important mid-products in the production of methane; their

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

concentrations affected the efficiency of fermentation and their effected on methane yield and
methanogenic bacteria growth [28]. The results in Fig 4 showed that when the system go to steady state,
the VFA and alkalinity values were in the range 1,100 1,250 mg/ L and 2,600 2,850 mg/L.
Additionally, the VFA/Alk ratio was less than 0.4 and Alk/COD ratio was in the range of 0.5 - 0.65 when
the system went into a steady state. The suitable VFA for the anaerobic digestion must be had a maximum
of 2,000 mg/L and should have the VFA/Alk ratio less than 0.4 for the buffer capacity of a good system
and should have the Alk/COD ratio greater than 0.5 to keep the pH of the system decreased [26, 28].
700

(A)

5184
5000
4000
3000

2288
1945

2000

951
1000

Methane yield (mL CH4/gVS-added)

Cumulative methane production (mL CH4)

6000

(B)

577

600
500
400

278

300

211
200
88

100
0

T- MP

CSW-MP

GW -MP

T- MP

Syn -MP

CSW-MP

GW -MP

Syn -MY

3000

0.7

2500

0.6
0.5

2000

0.4
1500
0.3
1000
Total VFA(mg/L)
Alkalinity(mg/L)
VFA/Alk ratio
Alk/COD ratio

500
0
8

15

18

21
Time (day)

24

25

0.2

VFA/alkalinity ratio

Total VFA and alkalinity (mg/L)

Fig. 3. Methane production from anaerobic co-digestion of CSW with GW at ratio of 99:1; (A) cumulative methane production and
(B) methane yield: T-MP (Total methane production), CSW-MP (Canned seafood wastewater methane production), GW-MP
(Glycerol waste (1%) methane production and Syn-MY (Synergistic methane yield)

0.1
0.0

31

Fig. 4. Total VFA, alkalinity, VFA/alkalinity ratio and Alk/COD ratio at an OLR of 2 g COD/L. day in UASB reactor

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

70
Biogas(mL/day)
Methane (mL/day)
Methane(%)
pH

10000

60
50

8000

40
6000
30
4000

20

(A)

2000

10

Methane element (%) and pH

Biogas and methane production(mL/day)

12000

0
1 2 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 29 30 31

Time (day)
COD (in)

COD (out)

CODremove (%)

100
90

25000

80
70

20000

60
50

15000

40

10000

(B)

30

COD remove (%)

COD concentration (mg/L)

30000

20

5000

10
0

0
8

15

18

21
Time (day)

24

25

31

Fig. 5. (A) Biogas and methane production, methane element (%) and pH at an OLR of 2 g COD/L. day in UASB reactor (B) COD
(in), COD (out) and COD remove (%)

4. Conclusion
Anaerobic co-digestion of CSW with 1%GW had potential to improve the quality and quantity of
biogas. The cumulative methane production and methane yield were 5,184 mLCH4 and 577
mLCH4/g VS-added. The cumulative methane production and methane yield was increased by 167% and
108% when compared with digested CSW alone. The results also showed that the methane production in
continuous reactor was achieved with methane production rate of 2.33 LCH4/L-reactor.day and methane
yield was 309 mLCH4/g COD-removed (640 mLCH4/g VS-added) of removal COD 80%. Adding GW as
a carbon source into CSW was resulted in increasing C/N ratio from 11 to 26 and also diluted the toxicity
of ammonia in the system. The maximum methane production of co-digestion between 99%CSW and 1%
GW was 5.8 m3 CH4/ m3 of mixed wastewater and electricity production of 1 m3 of mixed wastewater
would be 207 MJ or 58 kWh of electricity. So, GW can be used a co-substrate because it enhanced a
potential the methane production in CSW.
Acknowledgements
I would like to thank the Office of the Higher Education Commission (OHEC) for support the funding
in this research.

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Kiattisak Panpong et al. / Energy Procedia 52 (2014) 328 336

References
[1] Palenzuela-Rollon A. Anaerobic Digestion of Fish Processing Wastewater with Special Emphasis on Hydrolysis of
Suspended Solids. Taylor and Francis 1999, London.
[2] Sunny N, Mathai PL. Physicochemical process for fish processing wastewater. International Journal of Innovative Research
in Science, Engineering and Technology 2013; 2(4): 901-905.
[3] Prasertsan P, Jung S, Buckle KA. Anaerobic filter treatment of fishery wastewater. World J. Microbiol. Biotechnol. 1994;
10: 1113.
[4] Chowdhury p, Viraraghavan T, Srinivasan A. Biological treatment processes for fish processing wastewater A review
.Bioresource Technology 2010; 10: 439449.
[5] Kayhanian M. (1999). Ammonia inhibition in high-solids biogasification: an overview and practical solutions.
Environmental Technology 1999; 20: 355365.
[6] Chen Y, Cheng JJ, Creamer KS. Inhibition of anaerobic digestion process: A review. Bioresource Technology 2008; 99(10):
4044-4064.
[7] Zeshan OP, Karthikeyan. Effect of C/N ratio and ammonia-N accumulation in a pilot-scale thermophilic dry anaerobic
digester. Bioresource Technology 2012; 113(0): 294-302.
[8] Guerrero L, Omil F, Mondez R, Lema JM. (1997). Treatment of saline wastewaters from fish meal factories in an anaerobic
filter under extreme ammonia concentrations. Bioresource Technology 1997; 61(1): 6978.
[9] Yazdani SS, Gonzalez R. Anaerobic fermentation of glycerol: A path to economic viability for the biofuels industry.
Current Opinion in Biotechnology 2007; 18: 213-219.
[10] Viana MM, Freitasb AV, Leitaoc RC, Pintoc GAS, Santaellad ST. (2012). Anaerobic digestion of crude glycerol: a
review. Environmental Technology Reviews 2012; 1(1): 8192.
[11] Ma J, Van Wam M, Carballa M, Verstraete W. Improvement of the anaerobic treatment of potato processing wastewater in
a UASB reactor by co-digestion with glycerol. Biotechnology Letter 2008; 30: 861867.
[12] Fountoulakis MS, Manios T. Enhanced methane and hydrogen production from municipal solid waste and agro-industrial
by-products co-digested with crude glycerol. Bioresource Technology 2009; 100: 3043-3047.
[13] lvarez JA, Otero L, Lema JM. A methodology for optimising feed composition for anaerobic co-digestion of agroindustrial wastes. Bioresource Technology 2010; 101(4):1153-1158.
[14] Astals S, Nolla-Ardvol V, Mata-Alvarez J. Anaerobic co-digestion of pig manure and crude glycerol at mesophilic
conditions: Biogas and digestate. Bioresource Technology 2012; 110(0): 63-70.
[15] Nuchdang S, Phalakornkule C. Anaerobic digestion of glycerol and co-digestion of glycerol and pig manure. Journal of
Environmental Management 2012; 101(0):164-172.
[16] Regueiro L, Carballa M, lvarez JA, Lema JM. Enhanced methane production from pig manure anaerobic digestion using
fish and biodiesel wastes as co-substrates. Bioresource Technology 2012; 123(0): 507-513.
[17] Hutnan M, Kolesarova N, Bodok I, Spalkova V. Lazor M. Possibilities of anaerobic treatment of crude glycerol from
biodiesel production. 36 th International Conferrece of Slovak Society of Chemical Engineering 2009; 156: 1-13.
[18] APHA. Standard methods for the examination of water and wastewater 22th edn, American Public Health Association,
Washington DC, USA; 2012.
[19] Hniman A, O-Thong S, Prasertsan P. Developing a thermophilic hydrogen producing microbial consortia from geothermal
spring for efficient utilization of xylose and glucose mixed substrates. Int J Hydrogen Energ 2011; 36: 8785-8793.
[20] O-Thong S, Boe K, Angelidaki I. Thermophilic anaerobic co-digestion of oil palm empty fruit bunches with palm oil mill
effluent for efficient biogas production. Applied Energy 2012; 93(0): 648-654.
[21] Symons GE, Buswell AM. The methane fermentation of carbohydrates. J. Am. Chem. Soc. 1993; 55(5): 20282036.
[22] Li Y, Park SY, Zhu J. Solid-state anaerobic digestion for methane production from organic waste. Renewable and
Sustainable Energy Reviews 2011; 15(1): 821-826.
[23] Astals S, Ariso M, Gal A, Mata-Alvarez J. Co-digestion of pig manure and glycerine: Experimental and modelling study.
Journal of Environmental Management 2011; 92(4): 1091-1096.
[24] Mladenovska Z, Dabrowski S, Ahring BK. Anaerobic digestion of manure and mixture of manure with lipids: Biogas
reactor performance and microbial community analysis. Water Science and Technology 2003; 48(6): 271-278.
[25] Fountoulakis MS, Petousi I, Manios T. Co-digestion of sewage sludge with glycerol to boost biogas production. Waste
Management 2010; 30(10): 1849-1853.
[26] Souza ME. Criteria for the utilization design and operation of UASB reactors. Water Science and Technology 1986;
18(12): 55-69.
[27] Khanal SK. Anaerobic Biotechnology for Bioenergy Production: Principles and Applications. Wiley-Blackwell 2008; p.4363.
[28] Wang J, Meng L. Effects of volatile fatty acid concentrations on methane yield and methanogenic bacteria. Biomass and
Bioenergy 2000; 33(5): 848-853.