Anopheles mosquito diversity, entomological indicators of malaria transmission and challenges of morphological identification in southwestern Ethiopia.

Background: A number of Anopheles species play either a primary or secondary role in malaria transmission. This necessitates understanding the species composition, bionomics, and behaviors of malaria mosquitoes in a particular geographic area, which is relevant to design and implement tailored intervention tools. This study aimed to assess the species composition, sporozoite infection rate, and blood meal origins of malaria mosquitoes in two malaria-endemic villages of Boreda district in Gamo Zone, southwest Ethiopia.

Methods: Thirty houses, 20 for Center for Disease Control and Prevention (CDC) light traps and 10 for Pyrethrum Spray Catches (PSC) were randomly selected for bimonthly mosquito collection from October 2019 to February 2020. An enzyme-linked immunosorbent assay (ELISA) was carried out to detect the blood meal origins and circumsporozoite proteins (CSPs). The entomological inoculation rate (EIR) was calculated by multiplying the sporozoite and human biting rates from PSCs. Anopheles gambiae complex and An. funestus group samples were further identified to species by the polymerase chain reaction (PCR). Anopheles species with some morphological similarity with An. gambiae complex or An. funestus group were tested using the primers of the two species complexes.

Results: A total of 14 Anopheles species were documented, of which An. demeilloni was found to be the dominant species. An. arabiensis was found to be positive for P. falciparum CSP with the overall CSP rate of 0.53% (1/190: 95% CI 0.01-2.9). The overall estimated P. falciparum EIR of An. arabiensis from PSC was 1.5 infectious bites/person/5 months. Of the 145 freshly fed Anopheles mosquitoes tested for blood meal sources, 57.9% (84/145) had bovine blood meal, 15.2% (22/145) had human blood meal origin alone, and 16.5% (24/145) had a mixed blood meal origin of human and bovine. Anopheles demeilloni were more likely to feed on blood meals of bovine origin (102/126 = 80.9%), while An. arabiensis were more likely to have blood meals of human origin. Eleven samples (2.6%; 11/420) were morphologically categorized as An. demeilloni, but it has been identified as An. leesoni (the only An. funestus group identified in the area) by PCR, though it requires additional verification by sequencing, because different species genes may have amplified for these species specific primers. Similarly, a small number of An. arabiensis were morphologically identified as An. salbaii, An. maculipalpis and An. fuscivenosus.

Conclusions And Recommendations: In spite of the wide variety of Anopheles mosquito species, An. arabiensis dominates indoor malaria transmission, necessitating additional interventions targeting this species. In addition, increasing entomological knowledge may make morphological identification less difficult.