Abstract

The mouth is an important niche for bacterial colonization. Previous research used mouth microbiota to predict diseases like colon cancer and inflammatory bowel disease (IBD). It is still unclear how the sampling methodology influences microbial characterization. Our aim was to determine if the sampling methods, e.g., cotton swab or tissue biopsy, and the age influence the oral microbial composition of mice. Microbial DNA was extracted using a commercial kit and characterized targeting the 16s rRNA gene from mouth swabs and tissue biopsies from 2 and 15 months old C57BL/6 male mice kept in the same SPF facility. Our results show statistical different microbial community of the different ages, type of sampling, and the two fixed factors age x type of sample (p-value < 0.05). At the genus level, we identified that the genera Actinobacillus, Neisseria, Staphylococcus, and Streptococcus either increase or decrease in abundance depending on sampling and age. Additionally, the abundance of Streptococcus danieliae, Moraxella osloensis, and some unclassified Streptococcus was affected by the sampling method. While swab and tissue biopsies both identified the common colonizers of oral microbiota, cotton swabbing is a low-cost and practical method, validating the use of the swab as the preferred oral sampling approach.

Highlights

  • Gastrointestinal microbiota has a strong relation with metabolism and the modulation of individual health [1]

  • As we found clear differences related to the sampling approaches and the age effect on oral microbiota, further research is required to understand how the bacterial ecology in the mouth fluctuates through the aging process and to establish a protocol that can be comparable between research groups

  • This study suggests that the sampling method is a factor to consider when determining bacterial abundances and diversity in the oral cavity

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Summary

Introduction

Gastrointestinal microbiota has a strong relation with metabolism and the modulation of individual health [1]. The human oral cavity is colonized by more than 700 bacterial species, making it the second most diverse site after the colon [3]. The combination of the mucosal shredding surfaces from the tongue, internal cheeks, and the hard tooth surfaces creates different environments for the adherence of bacteria in the mouth [4]. The biofilms in the oral cavity vary in bacterial composition and abundance, making the mouth a polymicrobial niche [5]. Several aspects can influence the composition of the oral microbiota, as the mouth is in direct contact with the exterior. The salivary flow and composition changes, the cellular exchange modifies, and the loss of dental pieces is frequent [7,8], being those aspects that could influence the adherence and growth of different bacterial species

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