MICROBIAL DIVERSITY OF CULICOIDES REEVESI FROM CHIHUAHUA, MEXICO: A METAGENOMIC ANALYSIS OF RRNA 16S

Abstract

This study aimed to investigate the microbial diversity of bacteria in the composite microbial community associated with Culicoides reevesi biting midges from Buenaventura municipality in the state of Chihuahua, Mexico, using a Sanger sequencing 16s rRNA metagenomics approach. Adult females of Culicoides reevesi were collected by human landing catches in the rainy season of 2023 and morphologically identified. They were grouped into pools of 25 individuals from which genomic DNA (gDNA) was extracted. Sanger sequencing of 16S rRNA was performed for a total of 4 pools, and the amplicon sequencing of the V3-V4 hypervariable region was done on Illumina Mseq platform to detect bacterial communities. The bioinformatic analysis included quality assessment, taxonomic classification, and visualization. The evaluation of the microbial community involved assessing taxa abundance and diversity using Mothur and QIIME2 software included in Galaxy Tool Shed (https://usegalaxy.eu/). Our study presents, for the first time in México and worldwide, an in-depth analysis of the bacteriome composition in C. reevesi, utilizing a 16S rRNA metagenomic approach. We emphasize the prevalence of dominant bacterial phyla, particularly Proteobacteria, alongside varying abundances of Actinobacteria, Firmicutes, Acidobacteria, and Bacteroidota, with a notable occurrence of Tenericutes. We identified intriguing species of both human and animal pathogenic bacteria. Moreover, we observed the absence of unidentified bacterial sequences, alongside the presence of other bacterial groups associated with the environment or plants. This has implications for both healthcare and ecological management, potentially simplifying control measures but also posing risks if the dominant species are harmful. This research enhances our understanding of the microbiome associated with Culicoides species, such as Culicoides reevesi, underscoring the need for further investigation to fully grasp their ecological importance and impact on public health.