Isolation and characterization of midgut microbiota in Anopheles mosquitoes from areas under varying insecticide pressure and malaria transmission settings of Ethiopia
- 1Department of Biology, College of Natural Sciences, Jimma University
- 2School of Medical Laboratory Sciences, Faculty of Health Sciences, Jimma University
Abstract
Investigating the composition and diversity of Anopheles midgut microbiota is essential for understanding vector–pathogen interactions, insecticide resistance, and malaria transmission dynamics across diverse eco-epidemiological settings. This study characterized culture-dependent midgut microbiota of Anopheles mosquitoes collected between July 2021 and March 2023 from three Ethiopian sites (Lare, Asendabo, and Batu), representing differing insecticide use intensity and malaria transmission risk. Midgut microbiota of adult female Anopheles funestus, An. coustani, An. pharaensis, and both larvae and adult Anopheles gambiae s.l. were analysed using culture-dependent methods. Microbiota from insecticide-survived and susceptible An. gambiae s.l. were also examined under semi-field conditions. Generalized linear models (GLMs) with a negative binomial distribution and log link were used to assess the effects of mosquito species, site, and life stage on midgut bacterial count and composition. A total of 129 mosquito specimens were dissected, yielding 659 bacterial and 5 fungal isolates. Eleven bacterial genera were detected, dominated by Staphylococcus (32%), Klebsiella (17.5%), Proteus (11.4%), and Bacillus (11.7%). Anopheles species appear to be a key driver of midgut microbiota abundance (Wald χ² = 8.33, df = 3, p = 0.040), with insecticide survivors also showing numerically higher (63%, n = 113) culturable bacterial count and genus richness along with site-level differences as compared to bacterial counts recorded from the susceptible counterparts (37%, n = 66). An. gambiae s.l. harboured over 70% of bacterial counts, with larvae also showing high culturable colonization intensity. Bacterial load and diversity with Shannon index showed small differences and may not be biologically meaningful as no statistical comparison of diversity indices was performed. The composition and abundance of culturable midgut microbiota were influenced more by mosquito species than by environmental variation, with larvae and adults of An. gambiae s.l. harbouring relatively higher number of culturable fraction of bacterial isolates. This study provides baseline information on culturable midgut microbiota of Anopheles mosquitoes in malaria-endemic areas of Ethiopia and highlights the need for integrated culture-dependent and sequencing-based studies to better understand their functional roles and potential applications in malaria control.
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BibTeX
@article{woyessa2026isolation,
title = {{Isolation and characterization of midgut microbiota in Anopheles mosquitoes from areas under varying insecticide pressure and malaria transmission settings of Ethiopia}},
author = {Delelegn Woyessa and Lijalem Hailu and Delenasaw Yewhalaw},
journal = {Scientific Reports},
year = {2026},
month = jul,
doi = {10.1038/s41598-026-62586-0},
url = {https://doi.org/10.1038/s41598-026-62586-0},
note = {CC BY},
}
RIS
TY - JOUR
TI - Isolation and characterization of midgut microbiota in Anopheles mosquitoes from areas under varying insecticide pressure and malaria transmission settings of Ethiopia
AU - Delelegn Woyessa
AU - Lijalem Hailu
AU - Delenasaw Yewhalaw
PY - 2026
DA - 2026/07/01
JO - Scientific Reports
DO - 10.1038/s41598-026-62586-0
UR - https://doi.org/10.1038/s41598-026-62586-0
AB - Investigating the composition and diversity of Anopheles midgut microbiota is essential for understanding vector–pathogen interactions, insecticide resistance, and malaria transmission dynamics across diverse eco-epidemiological settings. This study characterized culture-dependent midgut microbiota of Anopheles mosquitoes collected between July 2021 and March 2023 from three Ethiopian sites (Lare, Asendabo, and Batu), representing differing insecticide use intensity and malaria transmission risk. Midgut microbiota of adult female Anopheles funestus, An. coustani, An. pharaensis, and both larvae and adult Anopheles gambiae s.l. were analysed using culture-dependent methods. Microbiota from insecticide-survived and susceptible An. gambiae s.l. were also examined under semi-field conditions. Generalized linear models (GLMs) with a negative binomial distribution and log link were used to assess the effects of mosquito species, site, and life stage on midgut bacterial count and composition. A total of 129 mosquito specimens were dissected, yielding 659 bacterial and 5 fungal isolates. Eleven bacterial genera were detected, dominated by Staphylococcus (32%), Klebsiella (17.5%), Proteus (11.4%), and Bacillus (11.7%). Anopheles species appear to be a key driver of midgut microbiota abundance (Wald χ² = 8.33, df = 3, p = 0.040), with insecticide survivors also showing numerically higher (63%, n = 113) culturable bacterial count and genus richness along with site-level differences as compared to bacterial counts recorded from the susceptible counterparts (37%, n = 66). An. gambiae s.l. harboured over 70% of bacterial counts, with larvae also showing high culturable colonization intensity. Bacterial load and diversity with Shannon index showed small differences and may not be biologically meaningful as no statistical comparison of diversity indices was performed. The composition and abundance of culturable midgut microbiota were influenced more by mosquito species than by environmental variation, with larvae and adults of An. gambiae s.l. harbouring relatively higher number of culturable fraction of bacterial isolates. This study provides baseline information on culturable midgut microbiota of Anopheles mosquitoes in malaria-endemic areas of Ethiopia and highlights the need for integrated culture-dependent and sequencing-based studies to better understand their functional roles and potential applications in malaria control.
SN - 2045-2322
ER -