Gut microbiome Profiling of Healthy and Type 2 Diabetic Groups in Nagaland Using High-throughput Sequencing

Sengupta, Swapnil (2025) Gut microbiome Profiling of Healthy and Type 2 Diabetic Groups in Nagaland Using High-throughput Sequencing. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Swapnil Sengupta (20MS011)) 20MS011_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (3MB)

Abstract

India is regarded as the ‘diabetes capital of the world’, having the second most diabetic population in the world. Its prevalence is increasing rapidly, and recent studies have found out significant role of the human gut microbiota in the incidence of Type 2 Diabetes (T2D). However, most of these studies have been conducted outside of India, making it essential to explore the association in the Indian context. This is necessary to understand how the changes in gut microbiota can contribute to T2D in Indian population to address this growing crisis. We chose Nagaland state of India as our study site as it has a collection of the most ethnic tribes. As a part of the investigation, initially a survey was done in a few sites of the state focussing on various demographics and lifestyle factors to assess the risk associated with T2D. After finding some weak associations, we shifted our focus to the gut microbiome study where we collected samples from 24 individuals. 13 of the samples were from healthy individuals and 11 of them were from T2D patients. After extraction of the DNA from samples, third generation sequencing platform Illumina Miniseq was used for our study using metagenomic sequencing. Alpha diversity between the two groups showed an enrichment of the bacterial microbial diversity in the T2D patients. Beta diversity analysis using the Bray-Curtis distance showed not much of a difference between healthy and T2D gut. Furthermore, taxonomic profiling at the phylum, family and genus level revealed significant differences between the gut composition of the two groups. Indicator species analysis helped us to identify 12 species associated with T2D gut and 3 species associated with the healthy gut of the individuals of Nagaland. Our study also compared the results from Nagaland with three other Indian States-West Bengal, Jharkhand and. Significant differences and similarities were found in the gut compositions across the population. A total of 80 Indian samples were analysed overall where genera associated with Nagaland (positively and negatively), T2D, Healthy and older individuals were found. Our data from Nagaland was also globally compared with Indonesia for identifying similarities and differences. Finally, we did a fixative analysis between 95% ethanol and RNAprotect® for fixing our samples. Our study revealed significant bias created by the latter one and hence 95% ethanol emerged as a better fixative for further studies

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