Bioresource Technology 95 (2004) 1924

Anaerobic batch co-digestion of sisal pulp and sh wastes

Anthony Mshandete
b

a,b

, Amelia Kivaisi

a,*

, Mugassa Rubindamayugi a, Bo Mattiasson

a Applied Microbiology Unit, Department of Botany, University of Dar es Salaam, P.O. Box 35060, Dar es Salaam, Tanzania 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 sh waste was studied both with the wastes separately and with mixtures in various proportions. While the highest methane yields from sisal pulp and sh 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 sh 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 5994% in the methane yield as compared to that obtained from the digestion of pure fractions at 5% TS. 2004 Elsevier Ltd. All rights reserved.
Keywords: Anaerobic co-digestion; Batch; Sisal pulp; Fish waste; Sisal waste sludge

1. Introduction 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 rst severely inhibited and even may die. Therefore, it is important to have buering capacity in the system, i.e. products that will counteract the eects of the VFA need also to be formed. It is known that carbohydrate-rich substrates are good producers of VFA and that protein-rich substrates are yielding good buering capacity (Nyns, 1986; Mata-Alvarez et al., 2000). Despite, these co-digestion benets, it is not clear whether some wastes have adverse eects when used in conjunction with another waste
* Corresponding author. Tel.: +255-22-41-0223; fax: +255-22-2410078. E-mail address: akivaisi@amu.udsm.ac.tz (A. Kivaisi).

(Callaghan et al., 2002). However, digestion of mixtures of dierent wastes is seldom reported (De Baere, 2000). Based on this information it seems realistic to investigate the eects 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 sh processing industries along lake Victoria in Mwanza City alone produce 1600 tons of sh wastes per annum (UNDP, 1993). Currently these wastes are disposed o 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 anaerobic 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 emission. This is thus in line with Kyoto Summit agreement (Mata-Alvarez et al., 2000). For sustainable energy development in the developing world, low cost low 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 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.biortech.2004.01.011

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A. Mshandete et al. / Bioresource Technology 95 (2004) 1924 Table 1 Characteristics of pure fractions of sisal pulp and sh waste used in anaerobic degradation trials (mean values, n 3) Analyses pH Partial alkalinity (PA) (mg CaCO3 /l) Total alkalinity (TA) (mg CaCO3 /l) Total solids (TS) (% of fresh sample) Volatile solids (VS) (% of TS) Organic carbon (OC) % (dry wt) Total nitrogen (TN) % (dry wt) Total lipids % (dry wt) C:N Sisal pulp 5.6 9 87.5 49 1.08 5.7 45 Fish waste 6.9 530 2280 32.2 55.3 51 5.85 12.6 9

simplest designs and being least expensive for use as solid waste digesters hold great promises. However, there seems to be insucient knowledge on the conversion of substrates other than the traditionally used cow dung (Foresti, 2001). In this context, knowledge on solids content and waste to inoculum ratios are important. These are variables that aect the experimental assessment of the potential biodegradability of organic matter. The solids content is one of the parameters that has a huge impact on the cost, performance and reliability of the digestion process (Lissens et al., 2001). 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 fermentation process, notably due to lipids hydrolysis (Amon et al., 2001). In the literature, co-digestion of organic fractions with cow dung or with sewage sludge is frequently reported. This paper, therefore, reports for the rst time, the results on comparison of anaerobic batch digestion of sisal pulp and sh wastes separately as well as the co-digestion of both substrates.

sludge (SWS) was quantied using acetate and formate as carbon source. SWS was collected from the sedimentation pond at a depth of about 3 m, using a VANDORN grab sampler, at Ubena Zomozi sisal factory. SWS (TS of 10.9% out of which 52.4% being VS) was used as an inoculum for digester start up. 2.3. Batch bioreactors The biodegradability and co-digestion of sisal pulp and sh wastes were tested in 1000 ml bioreactors constructed by using conical glass asks 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 bioreactors were tted with gas sampling septa closed with n-butyl stoppers and sealed with aluminium caps. Initially, the digestion mixtures were ushed with nitrogen for 5 min to replace the air (oxygen) in order to achieve anaerobic conditions. Subsequently, the mouth of bioreactors 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

2. Methods 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 sh waste (FW) obtained from the landing beach in Dar es Salaam City in Tanzania, consisted of oals, 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, respectively. 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

Digestion of fresh sisal pulp or sh wastes, separately, was carried out at dierent wet biomass % (v/v) between 5 and 60. The waste to inoculum ratios were between 0.051.6 and 0.092.5 g VS waste/g VS inoculum, for sh waste and sisal pulp, respectively. The experimental set-

Table 2 Composition of the sisal pulp and sh waste combinations (% of wet weight) used in co-digestion trials (mean values, n 3) % Wet weight TS % VS % of TS OC % (dry wt) TN % (dry wt) Lipids % (dry wt) C:N ratio 50(FW):50(SP) 20.6 61.8 49.7 4.0 7.3 12 33(FW):67(SP) 16.6 67.6 46.6 2.8 4.8 16 25(FW):75(SP) 14.8 71.4 43.7 2.3 4.2 18 20(FW):80(SP) 13.6 73.6 38.7 1.7 3.8 23

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up for the pure sisal pulp and sh wastes consisted of 30 batch bioreactors for each substrate. A control bioreactor 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 sh waste, respectively. These experiments were terminated when no signicant biogas production was observed over a two-week period. In the co-digestion experiment, mixtures of FW and SP in varying proportions were digested in 15 bioreactors. The sh waste solid content at which the highest methane yield was obtained (5% of TS) was kept constant 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 sh 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 Ergder u 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, propionate and butyrate were analyzed by using a HewlettPackard gas chromatograph (type HP 5890). Samples were centrifuged, 3 ml of the ltrate was acidied with 20 ll conc. H2 SO4 and stored at )20 C. Before analysis the samples were thawed and ltered using 0.45 lm lters. Samples of 0.9 ml were mixed with 0.1 ml of 100 mM isobutyric acid as an internal standard. Subsequently, 0.25 ml 20% ortho-phosphoric acid was added and, after mixing thoroughly, the samples were allowed to stand for 30 min. Samples of 0.10.2 ll were injected into the glass column (1.8 m long and 2 mm internal diameter) lled with 10% SP1200/1% H3 PO4 on 80/ chromosorbWAW. Nitrogen was used as a carrier gas at a ow 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 Bjrnsson o et al. (2000) using a TIM titration manager with an ABU 901 Autoburette (Radiometer, Copenhagen, Denmark). 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

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 described by Nelson and Sommers (1996).

3. Results and discussion 3.1. Biodegradability of pure sh and sisal pulp wastes The extent on conversion of pure sh waste and sisal pulp at various substrate concentrations in terms of methane yield is shown in Fig. 1. Analysis of variance (ANOVA) for each sh and sisal pulp fractions showed that there was signicant dierence 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 previously observed for dierent 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 1736 days (depending on the material). Batch biomethanation of leafy biomass, comparable to sisal pulp biomass, gave a methane yield in range of 0.2710.429 CH4 m3 /kg VS added (Zubr, 1986; Sharma et al., 1988). In the case of biomethanation of sh wastes per se, literature data on methane yield are scarce. Ahring et al. (1992) reported methane yield in the range of 0.4500.500 m3 CH4 /kg VS added from sh 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 eect of dierent total wet biomass content % (v/v) of sisal pulp and sh wastes on the methane yield. The sisal pulp had TS of 9% and sh waste TS of 32%. The values are means standard deviations (vertical bars, n 3rd deviations).

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5%, since the methane yield increased signicantly when the waste/inoculum ratio decreased from 1.6 to 0.05 for sh 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 sh 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 6067% methane content (El-Shinnawi et al., 1989). 3.2. Co-digestion of sisal pulp and sh wastes The total methane production and methane yield by the digestions of co-digested sh and sisal pulp wastes are shown in Table 3. Analysis of variance (ANOVA) for methane yield for the substrates combinations showed that there was signicant dierences among the combinations tested (p < 0:0001). The total methane production and methane yields varied between 0.380.77 l and 0.300.62 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 methane production. Compared to the methane yields for the pure sisal pulp and sh waste, respectively co-digestion of the sh 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 5994%. 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 sh oal 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 dierent proportions of fresh wt% which represents dierent % of TS of sh and sisal pulp wastes (mean values, n 3) % Wet weight 50 33 25 20 FW:50 FW:67 FW:75 FW:80 SP SP SP SP % TS 20.6 16.6 14.8 13.6 Total (CH4 ) 0.38 0.77 0.57 0.50 Yield CH4 m3 /kg VS 0.31 0.62 0.48 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% methane content in this work is closer to the range of 5060% which is normally obtained from conventional anaerobic 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 sh 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 ratios between 25 and 30 as being optimal. However, some investigators argue that the C/N of approximately between 1619 (Nyns, 1986) and 16.818 (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 performance using Parthenium, a terrestrial weed, as feedstock 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, 0.020.08 mM for propionic acid and 0.0080.05 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 nal pH values range was 7.37.7 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 6.47.6 in a normal functioning digester, beyond which a state of inhibition may occur resulting from toxic eects 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 nal range was 6700 7700 mg CaCO3 /l. The latter, demonstrated an increased partial alkalinity in the range of 4352% compared to the initial values before anaerobic digestion. This provided further evidence that the co-digestion of sh and

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sisal pulp proportions studied was successful. Previously, laboratory studies on mesophilic and thermophilic anaerobic sludge digestion reported a range of 20004000 mg CaCO3 /l partial alkalinity as being typical for properly operating digesters (Pohland and Bloodgood, 1963). The initial values reported in this study fall within this range. However, the nal values are 24 times higher than the reported values. This increase could be due to generation of NH during the 4 digestion of protein in sh waste which resulted in an increased digester buering capacity and hence stability of the digesters. This is an interesting cost-eective approach since no external buer sources were added.

4. Conclusions This study has shown that anaerobic digestion of pure sisal pulp and sh wastes is a feasible process. Furthermore, anaerobic co-digestion of sh waste and sisal pulp is a viable alternative for recovering energy in the form of biogas with 6065% methane content while at the same time abating environmental pollution. To the best of our knowledge anaerobic co-digestion of sh waste and sisal pulp is being reported for the rst time. The results also indicate that co-digestion with 33% sh 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 sh wastes at 5% of TS. Therefore, further research is planned to run a continuous stirred tank reactor to examine the eect of adding dierent sh and sisal pulp waste blends to the system digesting sisal wastewater sludge to gain more information of the possible scale up of the process.

Acknowledgements This work was supported by Swedish International Development Agency (SIDA) through the BIO-EARNproject and their nancial support is grate fully acknowledged.

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