Abstract
Midgut symbiotic bacteria are known to play fundamental roles in the biology of mosquitoes, however knowledge of midgut bacterial communities associated with mosquitoes is scanty due to limitation of the isolation techniques based on culturing. In this study, the composition and diversity of midgut bacteria in field collected and lab reared adult female Anopheles gambiae and Culex quinquefasciatus mosquitoes was explored using the Illumina sequencing. Deoxyribonucleic acid was isolated from the pooled midgut extracts and their 16S rRNA gene sequenced using Illumina sequencing platform. Operational taxonomic units (OTUs) were analyzed using QIIME 1.8.0; taxonomy was assigned using BLASTn against SILVA 119 and hierarchical clustering was done using R program software. Out of the total number of sequence reads obtained, 145 OTUs were realized at 3% genetic distance. The 145 OTUs spanned 12 phyla; Proteobacteria, Firmicutes, Cyanobacteria, Euryarchaeota, Gemmatimonadetes, Spirochaetae, Archeabacteria Verrucomicrobia, Chloroflexi, Bacteriodetes, Acidobacteria and Actinobacteria. Microbial community composition based on OTUs showed significant difference between field collected and lab reared mosquitoes (aµa2 = 45.0799, p = 3.2 × 10-5). Similarly, there was a significant difference in community composition at OTU level between Anopheles gambiae and Culex quinquefasciatus (aµa2 = 31.2257, p = 7.7 × 10-4). The bacterial composition and diversity appeared to be influenced by the environment and the species of the mosquitoes. Key words: Anopheles gambiae, Culex quinquefasciatus, midgut, DNA, diversity.
Highlights
Mosquitoes transmit diseases like malaria, dengue, lymphatic filariasis, yellow fever among others
There was a significant difference in community composition at Operational taxonomic units (OTUs) level between Anopheles gambiae and Culex quinquefasciatus (χ2 = 31.2257, p = 7.7 × 10-4)
The sequencing depth as shown by the rarefaction curve was exhaustive enough to ensure the inclusion of the entire diversity of the microbes in the midgut of the two species of mosquitoes collected from field and lab reared
Summary
Mosquitoes transmit diseases like malaria, dengue, lymphatic filariasis, yellow fever among others. Among these diseases, malaria is the most important mosquito borne disease with an estimated 214 million new cases of malaria worldwide (World Health Organization (WHO), 2015). The African region accounted for most of the global cases of malaria (88%), followed by South-East Asia region (10%) and Eastern Mediterranean region (2%) (WHO, 2015). In Kenya, there were an estimated 6.7 million new clinical cases and 4,000 deaths each year
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