Diversity of Culturable Bacteria Isolated From the Feces of Wild Anopheles darlingi (Diptera: Culicidae) Mosquitoes From the Brazilian Amazon

• Published in: Journal of medical entomology Vol. 58; no. 4; pp. 1900 - 1907
• Main Authors: Arruda, Andrelisse, Ferreira, Gabriel E. M., Júnior, Antônio Santos, Matos, Najla B., Carvalho, Tatiane S., Ozaki, Luiz S., Stabeli, Rodrigo G., Silva, Alexandre A. E.
• Format: Journal Article
• Language: English
• Published: US Entomological Society of America 01-07-2021; Oxford University Press
• Subjects: 16S rRNA; Analysis; Anopheles; Anopheles darlingi; Aquatic insects; Bacteria; Culicidae; DNA sequencing; Feces; Frequency distribution; Geographical distribution; Glycerol; Homology; Infectious diseases; Malaria; malaria vector; MALDI-TOF; Medical research; Medicine, Experimental; Microbiota; Microbiota (Symbiotic organisms); Microorganisms; Midgut; Mosquitoes; Nucleotide sequencing; phylogenetic analyses; Phylogeny; RNA; rRNA 16S; Rural areas; Species richness; Vector-borne diseases; VECTOR/PATHOGEN/HOST INTERACTION, TRANSMISSION; Brazil; microbiota; 16S rRNA; MALDI-TOF; malaria vector; phylogenetic analyses
• ISSN: 0022-2585; 1938-2928
• DOI: 10.1093/jme/tjab028

Microorganisms living in the midgut of Anopheles mosquitoes have been studied to fight vector-borne diseases, such as malaria. Studies on the microbiota of the Neotropical Anopheles darlingi, the most important Brazilian vector for malaria, have been reported for the same purpose. Our aims were to isolate and identify culturable bacteria from An. darlingi mosquito guts through their feces and to estimate the species richness and the frequency distribution of the sampled bacteria. Sixty wild females of An. darlingi mosquitoes were captured at two rural locations, near Porto Velho, Rondônia, Brazil. Bacteria were isolated from mosquito feces, which were collected using cages which permit the collection of feces on LB nutrient agar plates. Sixty bacterial colonies were isolated and stored in glycerol at –80°C. Bacteria were identified by sequencing their 16S rRNA gene obtained using PCR and Sanger sequencing. To aid in species identification, MALDI-TOF, VITEK2, and BBL Crystal were used as complementary protocols.The sequences obtained from the 60 bacterial isolates were compared to sequences deposited in GenBank (NCBI) using BLAST. Homology greater than 97% between the query and the subject was used as the criteria for assigning the identity of each isolate. Fourteen species from eight different genera were identified among the 60 isolates. The most frequent species were Serratia liquefaciens (20%) and Serratia marcescens (15%). Due to their established apathogenicity and according to previous studies, we suggest Serratia and Pantoea species as suitable for paratransgenesis development to fight malaria in Brazilian Amazon.