Shotgun Metagenomic Profiling of Phlebotomus papatasi (Diptera: Psychodidae, Phlebotomine) in Zoonotic Cutaneous Leishmaniasis Endemic Foci of Iraq: Microbiota, Resistome, and Virulome.

Abstract

This study aims to develop a regional baseline for future individual-level and comparative studies of microbial ecology and symbiont-based vector control. Phlebotomus papatasi is a major vector of Leishmania major and an important component of the transmission cycle of zoonotic cutaneous leishmaniasis (ZCL) in Iraq, although the internal microbial communities of this vector are poorly characterised in the region. In this study, shotgun metagenomic sequencing was employed to investigate the microbial communities of a pooled sample of 129 blood-fed female Ph. papatasi collected from ZCL-endemic foci in Nineveh Governorate, Iraq, from 2024 to 2025. Approximately 32.9 million raw reads were obtained from sequencing, with the quality-filtered, host-removed, and low-diversity-filtered reads analysed for taxonomic and functional profiles. The data revealed a complex microbial community inside blood-fed Ph. papatasi dominated by bacteria. Moreover, several species of Proteobacteria in blood-fed Ph. papatasi were detected, which accounted for 98.32% of the classified reads, with Enterobacter cloacae as the most common species (74.34% of classified reads). Strains associated with Wolbachia were also detected (approximately 8.96% of classified reads). The microbial data further revealed the presence of genes associated with antimicrobial resistance including oqxA/oqxB-associated efflux and AmpC-type β-lactamase families, and virulence factors associated with motility, iron acquisition, and secretion systems. Nevertheless, the data represented the distribution of sequencing reads within the pooled sample (single sample), rather than the prevalence or average abundance among individual sand flies.