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Background   Lutzomyia evansi , a phlebotomine insect endemic to Colombia’s Caribbean coast, is considered to be the main vector of visceral and cutaneous leishmaniasis in the region. Although insects of this species can harbor pathogenic and non-pathogenic microorganisms in their intestinal microbiota, there is little information available about the diversity of gut bacteria present in  Lutzomyia evansi . In this study, conventional microbiological methods and molecular tools were used to assess the composition of bacterial communities associated with  Lutzomyia evansi  guts in immature and adult stages of natural populations from the department of Sucre (Caribbean coast of Colombia).    Methods  Sand flies were collected from two locations (peri-urban and jungle biotype) in the Department of Sucre (Caribbean coast of Colombia). A total of 752  Lutzomyia evansi  intestines were dissected. In this study, 125 bacterial strains were isolated from different culture media (LB Agar, MacConkey Agar). Different methods were used for bacterial identification, including ribosomal intergenic spacer analysis (RISA) and analysis of the 16S rRNA and  gyr B gene sequences. The genetic profiles of the bacterial populations were generated and temporal temperature gradient gel electrophoresis (TTGE) was used to compare them with total gut DNA. We also used PCR and DNA sequence analysis to determine the presence of  Wolbachia  endosymbiont bacteria and  Leishmania  parasites.    Results  The culture-dependent technique showed that the dominant intestinal bacteria isolated belong to  Acinetobacter ,  Enterobacter ,  Pseudomonas ,  Ochrobactrum ,  Shinella  and  Paenibacillus  in the larval stage;  Lysobacter ,  Microbacterium ,  Streptomyces ,  Bacillus  and  Rummeliibacillus  in the pupal stage; and  Staphylococcus ,  Streptomyces ,  Brevibacterium ,  Acinetobacter ,  Enterobacter  and  Pantoea  in the adult stage. Statistical analysis revealed significant differences between the fingerprint patterns of the PCR-TTGE bands in bacterial communities from immature and adult stages. Additionally, differences were found in bacterial community structure in fed females, unfed females, males and larvae. The intestinal bacteria detected by PCR-TTGE were  Enterobacter cloacae  and  Bacillus thuringiensis , which were present in different life stages of  Lu. evansi , and  Burkholderia cenocepacia  and  Bacillus gibsonii , which were detected only in the larval stage.  Wolbachia  and  Leishmania  were not detected in gut samples of  Lutzomyia evansi .    Conclusions  The analyses conducted using microbiological and molecular approaches indicated significant variations in the bacterial communities associated with the gut of  Lu. evansi , depending on the developmental stage and food source. We propose that these elements affect microbial diversity in  L. evansi  guts and may in turn influence pathogen transmission to humans bitten by this insect.

Structural differences in gut bacteria communities in developmental stages of natural populations of Lutzomyia evansi from Colombia's Caribbean coast