In the recent study by Cirne (2006), it has beenreported that direct lipase enzyme addition and bioaug-mentation of anaerobic digester with a lipolytic anaerobicmicrobial strain enhanced lipid hydrolysis but theintermediate products (LCFAs) inhibited the latter stepsof biodegradation, lowering the overall efficiency of theprocess. It was also concluded that the bioaugmentingstrain was unable to compete with the “native” biomass ofthe digester (Cirne, 2006).Other studies on chemical pretreatment of lipid-richwastewaters revealed that addition of alkaline chemicalssuch as NaOH, KOH and Ca OH)
to slaughterhousewastewater and oil mill effluents improved hydrolysis ofsolid fatty residuals (Beccari et al., 1999; Masse et al.,2001; Mouneimne et al., 2003). Nevertheless, Masse andco-workers do not recommend pretreatment with an alkalibecause it results in an increase in pH in the bioreactor.Furthermore, results from pretreatment of lipid-richwastewaters by addition of calcium chloride and bento-nite clay indicated that these chemicals could reduce theinhibitory effects of LCFAs (Angelidaki et al., 1990;Broughton et al., 1998; Beccari et al., 1999). However,use of these chemicals was not recommended since theywork by limiting the concentration of dissolved LCFA thatsubsequently led to reduced methane yields. Lefebre etal. (1998) suggested the removal of large proportions oflipids (fats and grease) from the lipid-rich wastewatersbefore treatment but the study revealed that dissolvedand/or emulsified lipids can not be filtered off and gainaccess into treatment system (Cammarota et al., 2001).In addition, complete removal of oil and grease leads toloss of lipids that have very high methane productionpotential.A study by Callaghan et al. (1999) on batch co-digestion of waste organic solids reported that co-digestion of cattle slurry with fish offal and brewery solidsenhanced methane yield. Mshandete et al. (2004) alsoreported that anaerobic batch co-digestion of sisal pulpwith fish solid waste (wet weight proportions; 33% fishwaste: 67% sisal pulp) enhanced methane yield by 59-94% compared to sisal pulp and fish wastes alone.Furthermore, Ahring (2003) reported a two-fold increasein the yield of methane, from 25 to 50 m
cattle waste when fish oil (total concentration 5%) wasadded to manure digester. Ahring (2003) recommendedthat adding lipids to anaerobic digester to enhance theproduction of methane is a promising approach thatneeds continued exploration. Similarly, Cirne (2006)recommended that lipid-containing waste has a very highmethane production potential and therefore research onthe limiting steps of the conversion of lipids to methaneneed to be continued. Therefore, this study aimed at
investigating new alternative methods that can minimiseand/or alleviate inhibition caused by lipids and LCFAs, andenhance the anaerobic digestion of Nile perch fishprocessing wastewater (FPW) generated along Lake
Victoria. To the best of our know-ledge, this is the firstreport on enhancement of AD of FPW by co-digestion,Gumisiriza et al. 329physical and biological pre-treatment methods.
MATERIAL AND METHODSSubstrate and inoculum
The substrate investigated in this study was Nile perch fish pro-cessing wastewater (FPW) collected from a mixed stream at Vicfishlimited in Mwanza City on the southern shore of Lake Victoria. Theinoculum was collected from a stabilization pond receiving highstrength FPW at the same industry where the substrate was collec-ted. Brewery wastewater used in co-digestion experiment wassampled from a mixed stream at Tanzania breweries limited (TBL)in Mwanza City, Tanzania.
Bioreactor configuration and operation
Biogas production from FPW sample was investigated in 500 mlbatch bioreactors consisting of wide mouth Erlenmeyer conicalflasks with a working volume of 0.36 L. The bioreactors weredesigned and operated according to Mshandete et al
(2004). Themethane content and biogas volume were measured after every 72h. The biogas volume and methane content were determinedaccording to Ergüder et al. (2001).
Pretreatment of FPW by addition of microbial cultures
The microbial strains used in this study were two bacterial strains:CBR 11 and BR10 isolated from a local stabilization pond treatinghigh strength FPW at Vicfish industry in Mwanza city, Tanzania.The two strains exhibited lipolytic activity when cultured in TributyrinAgar (TBA) and broth media with lipid as the sole carbon source.The experiment consisted of two set-up: addition of separate straincultures and addition of a mixed culture of the two strains. Themicrobial cultures were added to the FPW according to Cirne(2006). The amount of the lipolytic strain added corresponded to1.3% VS of the inoculum added for a 1:1 inoculum-to-substrate gVSloading ratio. For each set up, the experiment consisted of thirteenaerobic incubation periods: 0, 3, 6, 9, 12, 15, 18, 21, 24, 36, 48, 60and 72 h. All batch bioreactors were incubated at 28
C whileshaking at 75 rpm. For addition of mixed cultures, 72-h brothcultures of the two bacterial strains were mixed in the ratio of 1:1. Atthe end of all the pretreatment periods, inoculum was added in aratio of 1:1 inoculum-to-substrate gVS. The experiment was set upin triplicate and was run for 36 days when biogas production hadceased.
Co-digestion of FPW with brewery wastewater
The brewery wastewater was added to 160 ml (0.534 gVS) of FPWat increasing %VS concentrations of: 0, 2, 4, 6, 8, 10, 12, 14, 16, 18and 20. Inoculum was added to each of the fractions including thecontrol at the ratio of 1:1 (inoculum gVS-to-substrate (FPW) gVS).The investigation was carried out in triplicate and was run for 36days when biogas production had ceased.
Pretreatment of FPW by LCFA removal
The bacterial strain (CBR11) which showed lipolytic activity withoutproteolysis, was cultured in Tributyrin broth medium. The lipaseenzyme was extracted according to the standard protocol describedby Bezerra et al. (2006).The broth culture media were centrifuged