Isolation and Characterization of Mucosa Associated Microbiota (MAM) of Stomach in Functional Dyspepsia Patients Among Pakistani Population

Abstract

Modern advancements in the sequencing technology for analyzing microbial communities have provided valuable insights on the intricate microbiota present in mammalian gastrointestinal tract whose composition have a significant impact in determining the overall health of the host. Functional dyspepsia (FD) is a prevalent chronic functional condition affecting the gastrointestinal system. Due to scarcity of the information of its pathogenesis, the treatment of functional dyspepsia poses challenges, and for the vast majority of patients, the problem persists chronically with symptoms that vary over time. Current study was designed for the characterization of mucosa associated microbiota (MAM) of stomach among functional dyspepsia patients. Biopsy samples of patients presenting symptoms of dyspepsia were collected from Holy Family Hospital Rawalpindi. Samples were processed and cultured repeatedly to isolate pure bacterial colonies. The pure isolates were then subjected to colony PCR for the amplification of 16srRNA gene using universal primers 27F and 1492R. For the identification of the isolates, Sanger Sequencing was done with 518F and 800R primers which cover the V4 region of the 16srRNA gene. Sequencing and Phylogenetic Analysis revealed that 41% of the isolates identified belong to phyla Firmicutes and 59% to Actinobacteria. Among Firmicutes most of the isolates belong to Lactobacillus, while among Actinobacteria most of isolates belongs to the family Micrococcaceae. The phylum Actinobacteria was found to be highly abundant in the analyzed samples. Thus, our results indicate high prevalence of Firmicutes and Actinobacteria in stomach microbiota of patients suffering from Functional Dyspepsia in Pakistan. Overall, the current study provides a better understanding of stomach microflora associated with functional dyspepsia. Exploration of the intricate relationship between the host and microbiota holds the potential to uncover innovative therapeutic strategies that specifically and effectively cure different aspects of the disease