Entomological survey of sibling species in the Anopheles funestus group in Tanzania confirms the role of Anopheles parensis as a secondary malaria vector.

BACKGROUND: Malaria transmission in Tanzania is driven by mosquitoes of the Anopheles gambiae complex and Anopheles funestus group. The latter includes An. funestus s.s., an anthropophilic vector, which is now strongly resistant to public health insecticides, and several sibling species, which remain largely understudied despite their potential as secondary vectors. This paper provides the initial results of a cross-country study of the species composition, distribution and malaria transmission potential of members of the Anopheles funestus group in Tanzania. METHODS: Mosquitoes were collected inside homes in 12 regions across Tanzania between 2018 and 2022 using Centres for Disease Control and Prevention (CDC) light traps and Prokopack aspirators. Polymerase chain reaction (PCR) assays targeting the noncoding internal transcribed spacer 2 (ITS2) and 18S ribosomal DNA (18S rDNA) were used to identify sibling species in the An. funestus group and presence of Plasmodium infections, respectively. Where DNA fragments failed to amplify during PCR, we sequenced the ITS2 region to identify any polymorphisms. RESULTS: The following sibling species of the An. funestus group were found across Tanzania: An. funestus s.s. (50.3%), An. parensis (11.4%), An. rivulorum (1.1%), An. leesoni (0.3%). Sequencing of the ITS2 region in the nonamplified samples showed that polymorphisms at the priming sites of standard species-specific primers obstructed PCR amplification, although the ITS2 sequences closely matched those of An. funestus s.s., barring these polymorphisms. Of the 914 samples tested for Plasmodium infections, 11 An. funestus s.s. (1.2%), and 2 An. parensis (0.2%) individuals were confirmed positive for P. falciparum. The highest malaria transmission intensities [entomological inoculation rate (EIR)] contributed by the Funestus group were in the north-western region [108.3 infectious bites/person/year (ib/p/y)] and the south-eastern region (72.2 ib/p/y). CONCLUSIONS: Whereas An. funestus s.s. is the dominant malaria vector in the Funestus group in Tanzania, this survey confirms the occurrence of Plasmodium-infected An. parensis, an observation previously made in at least two other occasions in the country. The findings indicate the need to better understand the ecology and vectorial capacity of this and other secondary malaria vectors in the region to improve malaria control.

Variations in household microclimate affect outdoor-biting behaviour of malaria vectors.

Background: Mosquito behaviours including the degree to which they bite inside houses or outside is a crucial determinant of human exposure to malaria. Whilst seasonality in mosquito vector abundance is well documented, much less is known about the impact of climate on mosquito behaviour. We investigated how variations in household microclimate affect outdoor-biting by malaria vectors, Anopheles arabiensis and Anopheles funestus. Methods: Mosquitoes were sampled indoors and outdoors weekly using human landing catches at eight households in four villages in south-eastern Tanzania, resulting in 616 trap-nights over 12 months. Daily temperature, relative humidity and rainfall were recorded. Generalized additive mixed models (GAMMs) were used to test associations between mosquito abundance and the microclimatic conditions. Generalized linear mixed models (GLMMs) were used to investigate the influence of microclimatic conditions on the tendency of vectors to bite outdoors (proportion of outdoor biting). Results:  An. arabiensis abundance peaked during high rainfall months (February-May), whilst An. funestus density remained stable into the dry season (May-August) . Across the range of observed household temperatures, a rise of 1 ºC marginally increased nightly An. arabiensis abundance (~11%), but more prominently increased An. funestus abundance (~66%). The abundance of An. arabiensis and An. funestus showed strong positive associations with time-lagged rainfall (2-3 and 3-4 weeks before sampling). The degree of outdoor biting in An. arabiensis was significantly associated with the relative temperature difference between indoor and outdoor environments, with exophily increasing as temperature inside houses became relatively warmer. The exophily of An. funestus did not vary with temperature differences.   Conclusions: This study demonstrates that malaria vector An. arabiensis shifts the location of its biting from indoors to outdoors in association with relative differences in microclimatic conditions. These environmental impacts could give rise to seasonal variation in mosquito biting behaviour and degree of protection provided by indoor-based vector control strategies.

Crowdsourcing Vector Surveillance: Using Community Knowledge and Experiences to Predict Densities and Distribution of Outdoor-Biting Mosquitoes in Rural Tanzania.

Lack of reliable techniques for large-scale monitoring of disease-transmitting mosquitoes is a major public health challenge, especially where advanced geo-information systems are not regularly applicable. We tested an innovative crowd-sourcing approach, which relies simply on knowledge and experiences of residents to rapidly predict areas where disease-transmitting mosquitoes are most abundant. Guided by community-based resource persons, we mapped boundaries and major physical features in three rural Tanzanian villages. We then selected 60 community members, taught them basic map-reading skills, and offered them gridded maps of their own villages (grid size: 200m×200m) so they could identify locations where they believed mosquitoes were most abundant, by ranking the grids from one (highest density) to five (lowest density). The ranks were interpolated in ArcGIS-10 (ESRI-USA) using inverse distance weighting (IDW) method, and re-classified to depict areas people believed had high, medium and low mosquito densities. Finally, we used odor-baited mosquito traps to compare and verify actual outdoor mosquito densities in the same areas. We repeated this process for 12 months, each time with a different group of 60 residents. All entomological surveys depicted similar geographical stratification of mosquito densities in areas classified by community members as having high, medium and low vector abundance. These similarities were observed when all mosquito species were combined, and also when only malaria vectors were considered. Of the 12,412 mosquitoes caught, 60.9% (7,555) were from areas considered by community members as having high mosquito densities, 28% (3,470) from medium density areas, and 11.2% (1,387) from low density areas. This study provides evidence that we can rely on community knowledge and experiences to identify areas where mosquitoes are most abundant or least abundant, even without entomological surveys. This crowd-sourcing method could be further refined and validated to improve community-based planning of mosquito control operations at low-cost.