Molecular and biochemical characterization of the oral microflora isolated from various regions of Nepal

Abstract

Formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment. Poor oral hygiene is one of the major causes for oral and dental diseases. And to understand the root cause of these oral diseases the identification of bacterial community is must. Along with oral community if biochemical mode of action of these microorganisms can be identified then possible mode of action towards situation of disease can be understood. So in this current work we try to obtain knowledge of microbial population and their biochemical mode of action among some of the Nepalese population using various molecular techniques to understand possible disease causing factor inside these oral cavity. Similarly, microbial enzymatic activity by these bacterial colonies was also explored. After the 16s rRNA sequencing 49 different organisms are identified having 19 different genera of oral microbial diversity in Nepalese population from various altitudes, ethnic groups, and various age groups. The bacterial isolates belonging to genus Enterococcus, Enterobacter, klebsiella, Bacillus, Pseudomonas, Staphylococcus, Serratia, Citrobacter, Stenotrophomonas, Achromobacter and their sub types are identified. GC-MS of bacterial extract provided the presence of 141 different compounds. Bacteria's with varying organic compounds secretion are obtained like alkyl, amines, esters phenols, chlorine and sulphur containing where these compounds either help them to establish formidably inside oral biofilm or help them to initiate and establish disease condition like oral malodors(sulphur compounds), carcinogenicity(acid and acid esters, chlorine compounds), caries(acid, acid eser). Escherichia marmotae the bacteria which was never isolated before in oral sample shows the anonymous nature in the oral cavity.The overall scientific data has paved the way to describe the distribution, pathogenic potential, physiology and variation of oral micro- organisms in Nepalese communities. Keywords: Biofilm, Enzyme assay, Sequencing, GC-MS