Isolation and identification of microflora from the midgut and salivary glands of Anopheles species in malaria endemic areas of Ethiopia.

BACKGROUND: Anopheles mosquitoes are of great importance to human health. A number 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 glands of Anopheles species. METHODS: A total of 20 pools (ten per pool) from insectary-reared and 56 pools (five per pool) of field-collected Anopheles mosquitoes were anesthetized by chloroform and dissected. 70% of ethanol was used for surface sterilization of mosquitoes and laboratory equipment, followed by rinsing Anopheles mosquitoes four times with 1X PBS. Each pool of dissected midgut and salivary gland sample was transferred in 1X PBS and squashed, incubated in the water bath and enriched in tryptic soya broth for 24 h at 35 ± 2 °C. As a control, the PBS solutions used to rinse the mosquitoes were also incubated in tryptic soya broth in the same conditions as the sample. After enrichment, a loopful of each sample was taken and inoculated on Blood, Chocolate, MacConkey, and Sabouraud Dextrose agar. Finally, the microbiota was isolated by colony characteristics, biochemical tests, and automated VITEK 2 Compact Analyzer. RESULTS: From all field and laboratory mosquitoes, Pseudomonas was found to be the dominant microbiota identified from all species of Anopheles mosquitoes. Acinetobacter and Klebsiellapneumonia and other families of gram-positive and gram-negative bacteria were identified. CONCLUSIONS: A number of bacteria were isolated and identified. This is the first report on isolation and identification of microbiota from midgut and salivary glands of Anopheles species in Ethiopia. It can be used as a baseline for studying the relationship between microbiota and mosquitoes, and for the development of a new malaria biological control.

Cutaneous leishmaniasis in Kutaber District, Ethiopia: Prevalence, sand fly fauna and community knowledge, attitude and practices.

Background: Cutaneous leishmaniasis (CL) affects 25% of the population living in the highlands of Ethiopia. CL intervention has not decreased the number of leishmaniasis patients. A cross-sectional study was conducted to determine CL prevalence, community's knowledge, attitude and practices (KAP), and the sand fly fauna in Kutaber district, northeast Ethiopia. Methods: A retrospective, community-based cross-sectional study was conducted in Boru Meda Hospital from December 2014-March 2021 to study CL prevalence of Kutaber district. A Pre-tested, well-structured questionnaire was used to collect data on the participants' socio-demographic characteristics, KAP towards CL and knowledge about sand fly vectors. Chi-square test and logistic regression analysis were used in the study, and data were analyzed using SPSS version 23 (p < 0.05). Results: A total of 10,002 (14.02%), of which 71,325 samples were confirmed as positive for CL. The infection rate of CL in females (7.1%) was a little bit higher than males (7.0%). More cases were recorded among 15-29 age category. The study also revealed that 77.1% of the respondents had poor knowledge about CL treatment, prevention, clinical presentation and disease transmission. Farmers tended to have poorer knowledge about sand flies than non-workers and students (32.7 vs. 35 and 44.1%; P = 0.049). Housewives had poorer knowledge about sand flies than farmers and workers (22.2 vs. 32.7 and 33.3%; P = 0.023). Phlebotomus longipes comprised the highest composition (80%) of the sand fly species identified in Kutaber district. Conclusions: The data showed that the community had poor knowledge about CL, vector, and transmission mode. CL preventive measures were prevalent, implying the need to raise CL awareness. Phlebotomus longipes was identified as the most dominant sand fly species which accounted for CL. The findings can be used in developing an effective control strategy to reduce CL transmission in the study area and elsewhere in Ethiopia.