Abstract

BackgroundSecondary symbionts of insects include a range of bacteria and fungi that perform various functional roles on their hosts, such as fitness, tolerance to heat stress, susceptibility to insecticides and effects on reproduction. These endosymbionts could have the potential to shape microbial communites and high potential to develop strategies for mosquito-borne disease control.Methodology/Principal findingsThe relative frequency and molecular phylogeny of Wolbachia, Microsporidia and Cardinium were determined of phlebotomine sand flies and mosquitoes in two regions from Colombia. Illumina Miseq using the 16S rRNA gene as a biomarker was conducted to examine the microbiota. Different percentages of natural infection by Wolbachia, Cardinium, and Microsporidia in phlebotomines and mosquitoes were detected. Phylogenetic analysis of Wolbachia shows putative new strains of Lutzomyia gomezi (wLgom), Brumptomyia hamata (wBrham), and a putative new group associated with Culex nigripalpus (Cnig) from the Andean region, located in Supergroup A and Supergroup B, respectively. The sequences of Microsporidia were obtained of Pi. pia and Cx. nigripalpus, which are located on phylogeny in the IV clade (terrestrial origin). The Cardinium of Tr. triramula and Ps. shannoni were located in group C next to Culicoides sequences while Cardinium of Mi. cayennensis formed two putative new subgroups of Cardinium in group A. In total were obtained 550 bacterial amplicon sequence variants (ASVs) and 189 taxa to the genus level. The microbiota profiles of Sand flies and mosquitoes showed mainly at the phylum level to Proteobacteria (67.6%), Firmicutes (17.9%) and Actinobacteria (7.4%). High percentages of relative abundance for Wolbachia (30%-83%) in Lu. gomezi, Ev. dubitans, Mi. micropyga, Br. hamata, and Cx. nigripalpus were found. ASVs assigned as Microsporidia were found in greater abundance in Pi. pia (23%) and Cx. nigripalpus (11%). An important finding is the detection of Rickettsia in Pi. pia (58,8%) and Bartonella sp. in Cx. nigripalpus.Conclusions/SignificanceWe found that Wolbachia infection significantly decreased the alpha diversity and negatively impacts the number of taxa on sand flies and Culex nigripalpus. The Principal Coordinate Analysis (PCoA) is consistent, which showed statistically significant differences (PERMANOVA, F = 2.4744; R2 = 0.18363; p-value = 0.007) between the microbiota of sand flies and mosquitoes depending on its origin, host and possibly for the abundance of some endosymbionts (Wolbachia, Rickettsia).

Highlights

  • A comprehensive understanding of the biology of insects requires that they must be studied in an ecological context where microorganisms are considered an important component [1], due to their essential role in disease transmission, and potential to develop strategies for mosquitoborne disease control [2]

  • Several studies have reported that multiple infections of Wolbachia-Cardinium occur commonly in several arthropod species (Drosophila simulans, Aedes albopictus, Nasonia vitripennis, Callosobruchus chinensis, Tetranychus cinnabarinus) [29,30,31,32], indicating that recently, we found multiple infection patterns in natural populations of Pintomyia evansi from Colombia in the intestinal microbiome [23], being the only report for sand flies

  • Conventional PCR used in this study could detect Wolbachia, Cardinium, and Microsporidia in total DNA extracted from sand flies and mosquitoes using reference primers (S1 Fig)

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Summary

Introduction

A comprehensive understanding of the biology of insects requires that they must be studied in an ecological context where microorganisms are considered an important component [1], due to their essential role in disease transmission, and potential to develop strategies for mosquitoborne disease control [2]. Some endosymbionts can be found in insect hemolymph and they can interact directly with secreted molecules of the humoral immune response In this regard, several studies have examined the interaction between endosymbionts infection in mosquitoes and their vector competence for important mammalian viral pathogens [4]. Secondary symbionts of insects include a range of bacteria and fungi that perform various functional roles on their hosts, such as fitness, tolerance to heat stress, susceptibility to insecticides, reproduction. These endosymbionts could have the potential to shape microbial communites and high potential to develop strategies for mosquito-borne disease control

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