Cellulolytic Activities of the Dung Beetle, Euoniticellus Intermedius, Larva Gut Micro-Flora

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Abstract

Background:

The life style and biology of dung beetles offer a significant opportunity for innovation in biofuel production. The larvae of the African dung beetle, Euoniticellus intermedius, feed solely on cow dung, eating and digesting the fibre while adults live on juices found in fresh dung. The larval gut system consists of a small, almost unrecognizable foregut and two distinct chambers; the midgut and hindgut. It is clear that these two chambers are the centres in which the dung material whose composition includes cellulose is processed. The goal of this study was to assess the cellulolytic activities of cultured gut micro-flora derived from E. intermedius, (Coleoptera: Scarabaeida).

Method:

Late second to third instar stage E. intermediuslarvae were dissected and the isolated gut micro-flora consortia aerobically cultured in media containing cellulose (filter paper) as the sole carbon source. Genomic DNA isolation was done on the gut consortia cultures after 10 days of culturing, using the ZR Fungal/Bacterial DNA MiniPrep kit (Zymo Research, USA). A complete and unbiased primary cosmid library was then constructed from the isolated genomic DNA using a cloning ready, pWEB-TNC™ Cosmid Cloning kit (EPICENTRE Biotechnologies, USA). The primary cosmid library clones were screened for endo-glucanase and cellobiohydrolase activities using Carboxymethyl Cellulose (CMC) and 4-Methylumbelliferyl-β-D-Cellobioside (MUC) plate assays respectively.

Results:

Results indicate that a total of 7 colonies out of 160 screened colonies showed positive CMC and MUC activities.

Conclusion:

This proves that E. intermediusis a potential source of cellulolytic micro-organisms and enzymes that can be used for cellulose derived biofuel production.

Related collections

Most cited references 27

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The diversity of the Insecta is reflected in the large and varied microbial communities inhabiting the gut. Studies, particularly with termites and cockroaches, have focused on the nutritional contributions of gut bacteria in insects living on suboptimal diets. The indigenous gut bacteria, however, also play a role in withstanding the colonization of the gut by non-indigenous species including pathogens. Gut bacterial consortia adapt by the transfer of plasmids and transconjugation between bacterial strains, and some insect species provide ideal conditions for bacterial conjugation, which suggests that the gut is a "hot spot" for gene transfer. Genomic analysis provides new avenues for the study of the gut microbial community and will reveal the molecular foundations of the relationships between the insect and its microbiome. In this review the intestinal bacteria is discussed in the context of developing our understanding of symbiotic relationships, of multitrophic interactions between insects and plant or animal host, and in developing new strategies for controlling insect pests.

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Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen.

R M Teather, P J Wood (1982)

The interaction of the direct dye Congo red with intact beta-D-glucans provides the basis for a rapid and sensitive assay system for bacterial strains possessing beta-(1 leads to 4),(1 leads to 3)-D-glucanohydrolase, beta-(1 leads to 4)-D-glucanohydrolase, and beta-(1 leads to 3)-D-glucanohydrolase activities. A close correspondence was observed between cellulolytic activity and beta-(1 leads to 4)-D-glucanohydrolase and beta-(1 leads to 4),(1 leads to 3)-D-glucanohydrolase activities in isolates from the bovine rumen. Many of these isolates also possessed beta-(1 leads to 3)-D-glucanohydrolase activity, and this characteristic may have taxonomic significance.