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Bacterial Ecology in Arthropod Digestive Tracts Casey Eisele Significance of Research Arthropods aid in ruminant digestion Little research has been conducted using arthropods Studies suggest that arthropods benefit from both – enzymatic activity (cellulose degradation) – products of fermentation Within the guts of Arthropods Microbes have been isolated from cockroaches, crickets, mealworms, desert millipedes, and scarab beetles Normal flora – high ratio of microbes/biomass – attachment to endothelium cockroaches: 100 species crickets: 25 species Materials and Methods Arthropods were sacrificed, sterilized and dissected Bacteria was liberated from the lining of the gut using a dilute solution of EDTA and sonication 1/10,000 dilutions were plated on – T Soy Agar Isolated bacteria were used to inoculate – Phenol Red Broth – Lactate Broth – Cellulose plates and broth Results Bacteria were isolated from all three arthropod species T Soy Agar (Periplaneta americana) Careful, they’re quick. Isolated 5 species – Underestimation – Psychotolerant T Soy Agar (Zophobas morio) Isolated 2 species T Soy Agar (Acheta dometicus) Isolated 2 species Populations and Diversity My Data Mealworm Cockroach 10 9 8 7 6 5 4 3 2 1 0 Weight of gut – – – Cricket (1x10 ^5 cfu/mg) Bacterial Count Bacteria/Biomass Species Cricket = 2mg Mealworm =54mg Cockroach =25mg # of species 2 2 >5 Bacteria/Biomass ratios confirmed the findings of Cazemier et al. and other studies Biochemical Tests 1) Fermentation Glycolytic Pathway host/bacteria bacteria Lactose (6 C) — —> Lactate (3 C) —> Pyruvate,Acetic Acid, Ethanol Phenol red broth Lactate Broth Results Bacteria were isolated from all three arthropod species 6 species fermented lactose Glycolytic Pathway Lactose Cockroach – – – – – Species 1 Species 2 Species 3 Species 4 Species 5 no proteolytic Mealworm – – —> Species 6 Species 7 Cricket – – Species 8 Species 9 proteolytic Lactate —> Pyruvate/Acetic/Ethanol Lactose Fermentation lactose fermenters control proteolytic proteolytic cricket mealworm cockroach Results Bacteria were isolated from all three arthropod species 6 species fermented lactose All Bacteria reduced lactate Glycolytic Pathway Lactose Cockroach – – – – – —> Lactate Species Species Species Species Species 1 2 3 4 5 no proteolytic Mealworm – Species 6 – Species 7 Cricket – Species 8 – Species 9 proteolytic —> Pyruvate/Acetic/Ethanol Biochemical Tests 1) Fermentation 2) Cellulose Degradation bacteria Cellulose —> Simple Sugars (6 C) Cellulose plate/broth Cellulose Degradation Cellulose tests were inconclusive Conclusions My study confirmed the presence of large bacterial populations living within the gut of arthropods These bacteria exhibited the ability to ferment compounds, whose end-products may be used by the host for nutrition Further testing would be needed to determine cellulose-degrading capabilities Acknowledgements Thank You – Professor Rose – Professor Smith – Tony