Bioresource Technology 95 (2004) 19–24

Anaerobic batch co-digestion of sisal pulp and fish wastes

a,b a,*
Anthony Mshandete , Amelia Kivaisi , Mugassa Rubindamayugi a, Bo Mattiasson b

a
Applied Microbiology Unit, Department of Botany, University of Dar es Salaam, P.O. Box 35060, Dar es Salaam, Tanzania
b
Department of Biotechnology, Center for Chemistry and Chemical Engineering, University of Lund, P.O. Box 124, S-22100 Lund, Sweden
Received 26 June 2003; accepted 27 January 2004
Available online 6 March 2004

Abstract
Co-digestion of various wastes has been shown to improve the digestibility of the materials and biogas yield. Batchwise digestion
of sisal pulp and fish waste was studied both with the wastes separately and with mixtures in various proportions. While the highest
methane yields from sisal pulp and fish waste alone were 0.32 and 0.39 m3 CH4 /kg volatile solids (VS), respectively, at total solid
(TS) of 5%, co-digestion with 33% of fish waste and 67% of sisal pulp representing 16.6% of TS gave a methane yield of 0.62 m3 CH4 /
kg VS added. This is an increase of 59–94% in the methane yield as compared to that obtained from the digestion of pure fractions at
5% TS.
Ó 2004 Elsevier Ltd. All rights reserved.

Keywords: Anaerobic co-digestion; Batch; Sisal pulp; Fish waste; Sisal waste sludge

1. Introduction (Callaghan et al., 2002). However, digestion of mixtures

Digestion of biomass in order to produce biogas and
biofertilizer is an attractive mode of treating waste
biomass. The process as such is very complex and it is
catalyzed by a consortium of microorganisms that in a
joined action convert complex macromolecules into low
molecular weight compounds such as methane, carbon
dioxide, water and ammonia. The composition of the
starting material is important in the sense that there is a
need for a suitable ratio between carbon and nitrogen.
Furthermore, main intermediates in the conversion are
volatile fatty acids (VFA). If a high concentration of
VFA is formed, pH will be reduced and that can reach
levels when the methanogenic bacteria are first severely
inhibited and even may die. Therefore, it is important to
have buffering capacity in the system, i.e. products that
will counteract the effects of the VFA need also to be
formed. It is known that carbohydrate-rich substrates
are good producers of VFA and that protein-rich sub-
strates are yielding good buffering capacity (Nyns, 1986;
Mata-Alvarez et al., 2000). Despite, these co-digestion
benefits, it is not clear whether some wastes have adverse
effects when used in conjunction with another waste
*
Corresponding author. Tel.: +255-22-41-0223; fax: +255-22-241-
0078.
E-mail address: akivaisi@amu.udsm.ac.tz (A. Kivaisi).
of different wastes is seldom reported (De Baere, 2000).
Based on this information it seems realistic to investigate
the effects of mixing biomass wastes when setting up an
anaerobic digestion process with the aim to convert the
biomass into biogas and biofertilizer.
In Tanzania, huge quantities of waste organic matter
are generated. Fifty-two sisal factories produce about
444,000 tons of sisal pulp annually. It has also been
estimated that fish processing industries along lake
Victoria in Mwanza City alone produce 1600 tons of fish
wastes per annum (UNDP, 1993). Currently these
wastes are disposed off untreated hence resulting in
environmental deterioration (Kivaisi and Mtila, 1998).
However, these wastes being abundant and rich in easily
biodegradable substrates, such as carbohydrates, lipids
and proteins are cheap potential feed stocks for anaero-
bic digestion in the production of biogas. It is important
to treat organic wastes under controlled conditions in to
order reduce spontaneous dissipation of CH4 to the
atmosphere. Moreover, production of biogas will reduce
the use of fossil fuels, thereby reducing the CO2 emis-
sion. This is thus in line with Kyoto Summit agreement
(Mata-Alvarez et al., 2000).
For sustainable energy development in the developing
world, low cost Ôlow tech’ renewable energy systems for
rural areas and peri-urban regions, have high potential
for application. To this end, batch systems having the

0960-8524/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2004.01.011
20 A. Mshandete et al. / Bioresource Technology 95 (2004) 19–24

simplest designs and being least expensive for use as Table 1

solid waste digesters hold great promises. However, Characteristics of pure fractions of sisal pulp and fish waste used in
anaerobic degradation trials (mean values, n ¼ 3)
there seems to be insufficient knowledge on the con-
version of substrates other than the traditionally used Analyses Sisal pulp Fish waste
cow dung (Foresti, 2001). In this context, knowledge on pH 5.6 6.9
Partial alkalinity (PA) (mg CaCO3 /l) – 530
solids content and waste to inoculum ratios are impor-
Total alkalinity (TA) (mg CaCO3 /l) – 2280
tant. These are variables that affect the experimental Total solids (TS) (% of fresh sample) 9 32.2
assessment of the potential biodegradability of organic Volatile solids (VS) (% of TS) 87.5 55.3
matter. The solids content is one of the parameters that Organic carbon (OC) % (dry wt) 49 51
has a huge impact on the cost, performance and reli- Total nitrogen (TN) % (dry wt) 1.08 5.85
Total lipids % (dry wt) 5.7 12.6
ability of the digestion process (Lissens et al., 2001).
C:N 45 9
Indeed, it has been suggested earlier by other workers
that the ratio of animal waste to other organic residues
must be optimized in order to avoid disturbances in the
sludge (SWS) was quantified using acetate and formate
fermentation process, notably due to lipids hydrolysis
as carbon source. SWS was collected from the sedi-
(Amon et al., 2001). In the literature, co-digestion of
mentation pond at a depth of about 3 m, using a
organic fractions with cow dung or with sewage sludge is
VANDORN grab sampler, at Ubena Zomozi sisal fac-
frequently reported. This paper, therefore, reports for
tory. SWS (TS of 10.9% out of which 52.4% being VS)
the first time, the results on comparison of anaerobic
was used as an inoculum for digester start up.
batch digestion of sisal pulp and fish wastes separately
as well as the co-digestion of both substrates.
2.3. Batch bioreactors

2. Methods The biodegradability and co-digestion of sisal pulp

2.1. Waste sources
Sisal pulp (SP), a leafy biomass waste produced
during sisal decortication, was obtained from a sisal
processing factory (Ubena Zomozi, Coast Region,
Tanzania). The fish waste (FW) obtained from the
landing beach in Dar es Salaam City in Tanzania, con-
sisted of offal’s, scales, gills and washing water. After
collection, FW was homogenized using a Moulinex
Masterchef 55 electronic kitchen blender and both SP
and FW were stored at )20 °C. Characteristics of the
pure SP, FW and mixed waste fractions used in the
experiments are described in Tables 1 and 2, respec-
tively.
2.2. Inoculum source
The lack of bulk anaerobic inocula in developing
countries like Tanzania has been cited as a bottle neck in
digester start up (Poggi-Varaldo et al., 1997). Therefore
in this study, methanogenic activity of sisal wastewater
and fish wastes were tested in 1000 ml bioreactors con-
structed by using conical glass flasks with a working
volume of 600 ml. The biogas produced was collected in
gas tight plastic tubings which were connected to gas
tight aluminium enforced polyethylene bags. The bio-
reactors were fitted with gas sampling septa closed with
n-butyl stoppers and sealed with aluminium caps. Ini-
tially, the digestion mixtures were flushed with nitrogen
for 5 min to replace the air (oxygen) in order to achieve
anaerobic conditions. Subsequently, the mouth of bio-
reactors were closed with n-butyl stoppers to ensure gas
tightness. The bioreactors were kept at an ambient
temperature of 27 ± 1 °C and were shaken manually for
1 min twice daily to mix their content.
2.4. Experimental set-up
Digestion of fresh sisal pulp or fish wastes, separately,
was carried out at different wet biomass % (v/v) between
5 and 60. The waste to inoculum ratios were between
0.05–1.6 and 0.09–2.5 g VS waste/g VS inoculum, for fish
waste and sisal pulp, respectively. The experimental set-

Table 2
Composition of the sisal pulp and fish waste combinations (% of wet weight) used in co-digestion trials (mean values, n ¼ 3)
% Wet weight 50(FW):50(SP) 33(FW):67(SP) 25(FW):75(SP) 20(FW):80(SP)
TS % 20.6 16.6 14.8 13.6
VS % of TS 61.8 67.6 71.4 73.6
OC % (dry wt) 49.7 46.6 43.7 38.7
TN % (dry wt) 4.0 2.8 2.3 1.7
Lipids % (dry wt) 7.3 4.8 4.2 3.8
C:N ratio 12 16 18 23
 A. Mshandete et al. / Bioresource Technology 95 (2004) 19–24
up for the pure sisal pulp and fish wastes consisted of 30
batch bioreactors for each substrate. A control biore-
actor containing only SWS (without waste) was included
and the biogas produced was subtracted from those
registered for the substrates used. All the digestions were
run in triplicates for 25 and 29 days for sisal pulp and
fish waste, respectively. These experiments were termi-
nated when no significant biogas production was ob-
served over a two-week period.
In the co-digestion experiment, mixtures of FW and
SP in varying proportions were digested in 15 bioreac-
tors. The fish waste solid content at which the highest
methane yield was obtained (5% of TS) was kept con-
stant in the preparation of four proportions namely,
50:50%, 33:67%, 25:75% and 20:80% (on wet weight
basis) of FW and SP, respectively. The volume of
inoculum added was kept constant at 590 ml (34 g VS),
the same volume used during digestion of fish waste at
5% TS. In this trial, digestions were run in triplicates for
24 days.
2.5. Analytical methods
The volume of biogas formed was measured by using
a graduated 100 ml gas tight plastic syringe with a
sample lock. The gas composition of 5 ml samples of the
biogas was estimated by the absorption of carbon
dioxide and hydrogen sulphide in concentrated alkaline
solution using serum bottles as described by Erg€ uder
et al. (2001). The bottles were shaken at 250 rpm for 4
min. In this method only CH4 was determined. Methane
yield was calculated as the net amount of methane
produced per unit VS added to the digester. Acetate, pro-
pionate and butyrate were analyzed by using a Hewlett-
Packard gas chromatograph (type HP 5890). Samples
were centrifuged, 3 ml of the filtrate was acidified with
20 ll conc. H2 SO4 and stored at )20 °C. Before analysis
the samples were thawed and filtered using 0.45 lm fil-
ters. Samples of 0.9 ml were mixed with 0.1 ml of 100
mM isobutyric acid as an internal standard. Subse-
quently, 0.25 ml 20% ortho-phosphoric acid was added
and, after mixing thoroughly, the samples were allowed
to stand for 30 min. Samples of 0.1–0.2 ll were injected
into the glass column (1.8 m long and 2 mm internal
diameter) filled with 10% SP1200/1% H3 PO4 on 80/
chromosorbWAW. Nitrogen was used as a carrier gas at
a flow rate of 40 ml/min. Oven, injection and detection
temperatures were, 130, 170 and 175 °C, respectively.
Partial alkalinity (PA), total alkalinity (TA) and pH,
were measured as previously described by Bj€ ornsson
et al. (2000) using a TIM titration manager with an
ABU 901 Autoburette (Radiometer, Copenhagen, Den-
mark). Samples were centrifuged at 6000 rpm for 3 min;
and 6 ml of the supernatant were used. TS and VS were
determined according to APHA Standard Methods
(APHA, 1995). Total nitrogen was determined by the
21
Kjeldahl method, total lipids were determined by Soxlet
extraction method using petroleum ether solvent
extraction, as described in APHA Standard Methods
(APHA, 1995). The organic carbon was determined by
rapid dichromate oxidation method previously de-
scribed by Nelson and Sommers (1996).
3. Results and discussion
3.1. Biodegradability of pure fish and sisal pulp wastes
The extent on conversion of pure fish waste and sisal
pulp at various substrate concentrations in terms of
methane yield is shown in Fig. 1. Analysis of variance
(ANOVA) for each fish and sisal pulp fractions showed
that there was significant difference in methane yield
when varying the % TS in the incubations (p < 0:0001).
Methane yield decreased with an increase in the total
solid substrate content. A similar tendency was previ-
ously observed for different types of animal, crop and
organic wastes (Badger et al., 1979; Itodo and Awulu,
1999). In this study the highest methane yield (m3 CH4 /
kg VS added) of 0.32 for SP and 0.39 for FW, were
obtained at 5% of TS after 25 and 29 days, respectively.
Similarly, Badger et al. (1979) reported highest methane
yield at 5% of TS when digesting various crops and
organic wastes in 11 batch-reactors during 17–36 days
(depending on the material). Batch biomethanation of
leafy biomass, comparable to sisal pulp biomass, gave a
methane yield in range of 0.271–0.429 CH4 m3 /kg VS
added (Zubr, 1986; Sharma et al., 1988). In the case of
biomethanation of fish wastes per se, literature data on
methane yield are scarce. Ahring et al. (1992) reported
methane yield in the range of 0.450–0.500 m3 CH4 /kg VS
added from fish oil sludge which is similar to what was
found in the present investigation. The ratio of waste/
inoculum was found to be a critical parameter especially
for incubations with a total solid content higher than
Methane yield CH4 m3 /Kg VS added
0.5
0.45
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0 5 10 15 20 25 30 35 40 45 50 55 60 65
Total wet biomass content %(v/v).
Sisal pulp Fish waste
Fig. 1. The effect of different total wet biomass content % (v/v) of sisal
pulp and fish wastes on the methane yield. The sisal pulp had TS of 9%
and fish waste TS of 32%. The values are means ± standard deviations
(vertical bars, n ¼ 3rd deviations).
22 A. Mshandete et al. / Bioresource Technology 95 (2004) 19–24
5%, since the methane yield increased significantly when
the waste/inoculum ratio decreased from 1.6 to 0.05 for
fish and 2.5 to 0.09 for sisal pulp, respectively. These
results are in line with those reported for conversion of
solid poultry slaughterhouse waste (Saliminen et al.,
2000 and Neves et al., 2002). The average approximate
methane content of the biogas increased with incubation
time and reached 59% for sisal pulp and 58% for fish
wastes at the end of digestion. Previous studies on batch
anaerobic digestion of rice straw, maize stalks, cotton
stalks and water hyacinth gave biogas with 60–67%
methane content (El-Shinnawi et al., 1989).
3.2. Co-digestion of sisal pulp and fish wastes
The total methane production and methane yield by
the digestions of co-digested fish and sisal pulp wastes
are shown in Table 3. Analysis of variance (ANOVA)
for methane yield for the substrates combinations
showed that there was significant differences among the
combinations tested (p < 0:0001). The total methane
production and methane yields varied between 0.38–0.77
l and 0.30–0.62 CH4 m3 /kg VS added, respectively with
values being highest for mixture containing 33%
FW:67% SP and lowest for 50% FW:50% SP. Generally,
based purely on total methane production and methane
yield among mixtures tested, the results suggest that, the
fraction with 33% FW:67% SP wet weight could be
suitable for successful co-digestion for enhanced meth-
ane production. Compared to the methane yields for the
pure sisal pulp and fish waste, respectively co-digestion
of the fish waste and sisal pulp at 33% FW:67% SP wet
weight proportions, with 16.6% of TS and a C:N ratio of
16, enhanced the methane yield by 59–94%. This could
be due to positive synergism in the digestion medium,
supplying missing nutrients and reducing/diluting of
inhibitory materials in feed stocks by the co-substrates
(Mata-Alvarez et al., 2000). This concurs with a recent
study on batch co-digestion of waste organic solids
which reported that fish offal and brewery solids mixed
with cattle slurry produced an enhancement in the
methane yield compared with that of a control digestion
of using cattle slurry alone (Callaghan et al., 1999).
Furthermore, Kaparaju et al. (2001) reported an
enhancement of about 60% in the methane yield with
Table 3
Total methane production and methane yield at different proportions
of fresh wt% which represents different % of TS of fish and sisal pulp
wastes (mean values, n ¼ 3)
% Wet weight % TS Total (CH4 ) Yield CH4 m3 /kg VS
50 FW:50 SP 20.6 0.38 0.31
33 FW:67 SP 16.6 0.77 0.62
25 FW:75 SP 14.8 0.57 0.48
20 FW:80 SP 13.6 0.50 0.44
co-digestion of industry confectionery waste with cow
manure.
The average CH4 content of the biogas produced
from 50:50 was (61%), 33:67 (64%); 25:75 (65%) and
20:80% (58%). The range between 60% and 65% meth-
ane content in this work is closer to the range of 50–60%
which is normally obtained from conventional anaero-
bic digestion of organic waste conducted in a single
stage-slurry digesters (Samani et al., 2001).
The C:N ratios of the co-digested sisal pulp and fish
wastes which ranged between 12 and 23 were within the
C:N ratios required for stable biological conversions
reported by others on anaerobic digestion of organic
wastes. Kayhanian and Hardy (1994) reported C:N ra-
tios between 25 and 30 as being optimal. However, some
investigators argue that the C/N of approximately be-
tween 16–19 (Nyns, 1986) and 16.8–18 (Kivaisi and
Mtila, 1998) are optimal for methanogenic performance
if poorly degradable compounds such as lignin are taken
into account. In addition, Gunaseelan (1995) suggested
a C:N of 11 being satisfactory for methanogenic per-
formance using Parthenium, a terrestrial weed, as feed-
stock for the digesters. Furthermore, Itodo and Awulu
(1999) reported successful anaerobic batch digestion for
poultry, cattle and piggery wastes slurries with a C:N of
6:1 and 9:1.
In a well balanced anaerobic digestion process, VFA
levels are low. In this study all the four combinations
examined showed lower levels of VFAs in their digested
slurry which ranged between 3.7 and 6.3 mM for acetic
acid, 0.02–0.08 mM for propionic acid and 0.008–0.05
mM for butyric acid. The initial propionic acid to acetic
acid ratio (P/A) at the beginning ranged between 0.007
and 0.01. It has been shown earlier by others examining
anaerobic digestion that increase in P/A ratio greater
than 1.4 and a build-up of acetic acid and butyrate to
above 200 mM as well as 100 mM of proprionate, can
explain process inhibition and ultimate digester failure
(Hill et al., 1987; Ahring et al., 1995).
The pH values and alkalinity before and after the
digestion trials indicate that the pH and alkalinity in
the digesters were conducive for biogas production. The
initial pH ranged between 7.7 and 7.8 while the final pH
values range was 7.3–7.7 which suggest that souring of
the digesters was not occurring in the co-digestion
mixtures tested. Similarly, Anderson and Yang (1992),
reported a range of pH 6.4–7.6 in a normal functioning
digester, beyond which a state of inhibition may occur
resulting from toxic effects of the hydrogen ions which
are believed to be closely related to the accumulation of
VFAs. The initial partial alkalinity ranged between 3600
and 3800 mg CaCO3 /l while the final range was 6700–
7700 mg CaCO3 /l. The latter, demonstrated an increased
partial alkalinity in the range of 43–52% compared to
the initial values before anaerobic digestion. This pro-
vided further evidence that the co-digestion of fish and
 A. Mshandete et al. / Bioresource Technology 95 (2004) 19–24
sisal pulp proportions studied was successful. Previ-
ously, laboratory studies on mesophilic and thermo-
philic anaerobic sludge digestion reported a range of
2000–4000 mg CaCO3 /l partial alkalinity as being typi-
cal for properly operating digesters (Pohland and
Bloodgood, 1963). The initial values reported in this
study fall within this range. However, the final values
are 2–4 times higher than the reported values. This in-
crease could be due to generation of NHþ 4 during the
digestion of protein in fish waste which resulted in an
increased digester buffering capacity and hence stability
of the digesters. This is an interesting cost-effective ap-
proach since no external buffer sources were added.
4. Conclusions
This study has shown that anaerobic digestion of
pure sisal pulp and fish wastes is a feasible process.
Furthermore, anaerobic co-digestion of fish waste and
sisal pulp is a viable alternative for recovering energy in
the form of biogas with 60–65% methane content while
at the same time abating environmental pollution. To
the best of our knowledge anaerobic co-digestion of fish
waste and sisal pulp is being reported for the first time.
The results also indicate that co-digestion with 33% fish
waste and 67% sisal pulp which represented 16.6% of TS
and a C:N ratio of 16 gave the highest methane yield of
0.62 m3 CH4 /kg VS added. This was an increase of 59–
94% in the methane yield compared to that obtained for
the digestion of pure sisal pulp and fish wastes at 5% of
TS. Therefore, further research is planned to run a
continuous stirred tank reactor to examine the effect of
adding different fish and sisal pulp waste blends to
the system digesting sisal wastewater sludge to gain
more information of the possible scale up of the pro-
cess.
Acknowledgements
This work was supported by Swedish International
Development Agency (SIDA) through the BIO-EARN-
project and their financial support is grate fully
acknowledged.
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