Project description:Bifidobacteria dominate the composition of the neonatal gut microbiota in the first number of weeks following birth. A number of species in particular are found with a significantly higher frequency in the microbiome of breastfed infants, owing to their ability to rely on Human Milk Oligosacchraides (HMOs) as their sole carbohydrate substrate; namely B. bifidum, B. longum spp. infantis and B. breve. Bifidobacterium kashiwanohense is a species that has been isolated previously only from the faeces of infants, but extremely infrequently at that. Relatively little is currently known about the species itself, let alone the metabolic pathways that allow it to successfully establish a population in the infant gut. We have isolated a novel strain of B. kashiwanohense from the faeces of a breastfed infant on the basis of its ability to utilise the HMO component fucosyllactose as its sole carbohydrate source. In this study, we read and annotate the full genome sequence of this novel strain, and use the data obtained to direct our further experimental analysis of fucosyllactose metabolism in B. kashiwanohense. Using transcriptomic and growth analysis results, we identify the genes responsible for B. kashiwanohense to utilise fucosyllactose, and employ a combination of cloning, in vitro hydrolysis assays, and further, recombinant transcriptomic and growth assays to elucidate the pathway for fucosyllactose metabolism in B. kashiwanohense, as well as revealing insight into fucosyllactose and fucose metabolism in Bifidobacteria as whole.
Project description:Faeces are considered easily accessible biological specimens that can be used to monitor gastrointestinal disorders. Extracellular vesicles (EVs) can be enriched from calf faeces, and these faecal extracellular vesicles (fEVs) consist of a diverse range of biological molecules, such as proteins, DNA, and RNA. The double membrane of EVs and the charged corona preserve their cargo, including proteins, from proteases, nucleases, and harsh environmental conditions. This enables the recovery of intact proteins secreted by the host and GI tract organisms, preserving their natural structure and function. While fEVs are suggested to mediate inter-species communication in the GI tract, the role of their protein cargo in GI infections remains unclear. Therefore, this study aimed to investigate the possibility of using fEVs as a non-invasive tool to investigate gut health and infections in claves. We considered Cryptosporidiosis as a model gut disease in calves to study the alteration in the proteomic profile of fEVs under gut infections.