Impact of cell lysis treatment before saliva metagenomic DNA extraction on the oral microbiome and the associated resistome

Author:

Tansirichaiya Supathep123ORCID,Songsomboon Kittikun4ORCID,Chaianant Nichamon5ORCID,Lertsivawinyu Wasawat1ORCID,Al‐Haroni Mohammed23ORCID

Affiliation:

1. Department of Microbiology, Faculty of Medicine Siriraj Hospital Mahidol University Bangkok Thailand

2. Department of Clinical Dentistry, Faculty of Health Sciences UiT the Arctic University of Norway Tromsø Norway

3. Centre for New Antimicrobial Strategies UiT the Arctic University of Norway Tromsø Norway

4. School of Life and Environmental Sciences The University of Sydney Sydney Australia

5. Faculty of Dentistry and Research Unit in Mineralized Tissue Reconstruction Thammasat University Pathumthani Thailand

Abstract

AbstractObjectivesThe human oral microbiome, a complex ecosystem linked to oral and systemic health, harbors a diverse array of microbial populations, including antimicrobial resistance genes (ARGs). As a critical component of the One Health approach to tackle antibiotic resistance, comprehending the oral resistome's composition and diversity is imperative. The objective of this study was to investigate the impact of chemical cell lysis treatment using MetaPolyzyme on the detectability of the oral microbiome, resistome, and DNA quality and quantity.Materials and MethodsSaliva samples were collected from five healthy individuals, and each of the samples was subjected to DNA extraction with and without the treatment with MetaPolyzyme. Through metagenomic sequencing, we analyzed, assessed, and compared the microbial composition, resistome, and DNA characteristics between both groups of extracted DNA.ResultsOur study revealed that MetaPolyzyme treatment led to significant shifts in the detectability of microbial composition, favoring Gram‐positive bacteria, notably Streptococcus, over Gram‐negative counterparts. Moreover, the MetaPolyzyme treatment also resulted in a distinct change in ARG distribution. This shift was characterized by an elevated proportion of ARGs linked to fluoroquinolones and efflux pumps, coupled with a reduction in the prevalence of tetracycline and β‐lactam resistance genes when compared with the nontreated group. Alpha diversity analysis demonstrated altered species and ARG distribution without affecting overall diversity, while beta diversity analysis confirmed significant differences in the taxonomical composition and oral resistome between treated and nontreated groups.ConclusionsThese findings underscore the critical role of cell lysis treatment in optimizing oral metagenomic studies and enhance our understanding of the oral resistome's dynamics in the context of antimicrobial resistance.

Publisher

Wiley

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