Analysis of euoniticellus intermedius, larva gut micro-flora: potential application in the production of biofuels.

Mabhegedhe, Munamato
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Recent years have seen a dramatic increase in first generation bio-fuel production, mainly driven by concerns of climate change and rising prices of transportation fossil fuels. Due to significant pressure on the few available food sources, second generation bio-fuels have entered the fray, as a sustainable alternative. This research‟s aim was to search for cellulolytic micro-organisms and enzymes from the gut of the dung beetle, Euoniticellus intermedius, (Coleoptera: Scarabaeida) that can be used in the production of second generation bio-fuels. Dung beetle larvae were dissected and the gut micro-flora cultured in cellulose medium. Bacterial growth and cellulase activity was monitored on a daily basis. DNA isolation was then done on the cellulose medium-cultured microbes and the isolated DNA cloned in E. coli. The clones were screened for cellulase activity using plate assays. A total of 7 colonies out of 160 screened colonies showed positive CMC (endo-β-1,4-glucanase) and MUC (cellobiohydrolase) activities. Sequencing of these positive colonies yielded mostly bacteria belonging to the Enterobacteriaceae family, most of which have not been previously reported to have cellulase activity. This study‟s findings prove that in addition to this dung beetle‟s gut being a fruitful source of microbial biodiversity, it is also a potential source of cellulolytic micro-organisms and enzyme activities that will aid the function and design of future bioreactors for the bio-fuel industry.