Isolation and Identification of Microflora From the Midgut and Salivary Glands of Laboratory Reared and Field Collected Anopheles Species from Some Malaria Endemic Areas of Ethiopia

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Addis Ababa University


Anopheles mosquitoes are of great importance to human health. They transmit pathogens including malaria parasites, filarial worms, and O'nyong-nyong and rift valley viruses. The numbers of studies have shown that, midgut and salivary gland microflora have an impact on malaria parasite burden through colonization mechanisms, involving either direct Plasmodium microbiota interaction or bacterial mediated induction of mosquito immune response. The objective of this study was to isolate and identify the microflora from the midgut and salivary gland of laboratory reared and field collected Anopheles species in some malaria endemic areas of Ethiopia. A total of twenty pools of mosquitoes, ten per pool (insectary), and ten pools, thirty per pool (field collected) were anesthetized by chloroform and dissected. 70% of ethanol was used for surface sterilization of mosquitoes followed by washing of each pool four times by 1x PBS and the environment. Each pool of dissected midgut and salivary gland samples was transferred in 3ml phosphate buffer saline, squashed and incubated in water bath until enriched in tryptic soya broth for 24hrs at 35±2oc. After enrichment, a loopful of each sample was taken and inoculated on Blood, Chocolate, Mac Conkey and Sabouraud dextrose agar for 24 hrs. at 35±2oc. Finally, the microbiota was isolated based on their colony characteristics and identified by conventional biochemical tests and automated VITEK 2 Compact Analyzer. All identified microbiota was stored in 20% tryptic soya broth with glycerol at -80 oc. From all field collected and insectaries, Pseudomonas 38 (29%) was found to be the dominant microbiota from all species. Anopheles gambiae s.l 77 (50.33%) had got the largest number of microbiota identified and An. arabiensis had identified diversified types of microbiota from the rest species. From this report, 40 genera of microbiota were identified and can be a milestone for studying relationship between microbiota and mosquitoes and for the development of a new malaria control strategy.



Anopheles Species, Identification, Malaria, Microflora, Midgut, Salivary Glands