Temporally consistent predominance and distribution of secondary malaria vectors in the Anopheles community of the upper Zambezi floodplain

Regional optimisation of malaria vector control approaches requires detailed understanding both of the species composition of Anopheles mosquito communities, and how they vary over spatial and temporal scales. Knowledge of vector community dynamics is particularly important in settings where ecohydrological conditions fluctuate seasonally and inter-annually, such as the Barotse floodplain of the upper Zambezi river. DNA barcoding of anopheline larvae sampled in the 2019 wet season revealed the predominance of secondary vector species, with An. coustani comprising > 80% of sampled larvae and distributed ubiquitously across all ecological zones. Extensive larval sampling, plus a smaller survey of adult mosquitoes, identified geographic clusters of primary vectors, but represented only 2% of anopheline larvae. Comparisons with larval surveys in 2017/2018 and a contemporaneous independent 5-year dataset from adult trapping corroborated this paucity of primary vectors across years, and the consistent numerical dominance of An. coustani and other secondary vectors in both dry and wet seasons, despite substantial inter-annual variation in hydrological conditions. This marked temporal consistency of spatial distribution and anopheline community composition presents an opportunity to target predominant secondary vectors outdoors. Larval source management should be considered, alongside prevalent indoor-based approaches, amongst a diversification of vector control approaches to more effectively combat residual malaria transmission.

High efficacy of microbial larvicides for malaria vectors control in the city of Yaounde Cameroon: a cluster randomised study

The rapid expansion of insecticide resistance and outdoor malaria transmission are affecting the efficacy of current malaria control measures. In urban settings, where malaria transmission is focal and breeding habitats are few, fix and findable, the addition of anti-larval control measures could be efficient for malaria vector control. But field evidences for this approach remains scarce. Here we provide findings of a randomized-control larviciding trial conducted in the city of Yaoundé that support the efficacy of this approach. A two arms random control trial design including 26 clusters of 2 to 4 km2 each (13 clusters in the intervention area and 13 in the non-intervention area) was used to assess larviciding efficacy. The microbial larvicide VectoMax®G combining Bacillus thuringiensis var israelensis (Bti) and Bacillus sphaericus in a single granule was applied twice per month in all standing water collection points. The biting anopheline density collected using CDC light traps was used as the primary outcome, secondary outcomes included the entomological inoculation rate, breeding habitats with anopheline larvae, and larval density. Baseline entomological data collection was conducted for 17 months from March 2017 to July 2018 and the intervention lasted 26 months from September 2018 to November 2020. The intervention was associated with a reduction of over 85% of habitats with anopheline larvae. The application of the larvicide also resulted in a reduction of 68% of adult anopheline biting density and of 79% of the entomological inoculation rate (OR 0.21; 95% CI 0.14–0.30, P < 0.0001). A reduction of 68.27% was recorded for indoor biting anophelines and 57.74% for outdoor biting anophelines. No impact on the composition of anopheline species was recorded. A reduction of over 35% of adult Culex biting densities was recorded. The study also assessed the impact of the microbial larvicide on non-target organisms and registered no significant impact of the larvicide VectoMax on the aquatic microfauna diversity. The study indicated high efficacy of larviciding for reducing malaria transmission intensity in the city of Yaoundé. Larviciding could be part of an integrated control approach for controlling malaria vectors and other mosquito species in the urban environment.

Mapping the breeding sites of Anopheles gambiae s. l. in areas of residual malaria transmission in central western Senegal

Despite the deployment of several effective control interventions in central-western Senegal, residual malaria transmission is still occurring in some hotspots. To better tailor targeted control actions, it is critical to unravel the underlying environmental and geographical factors that cause the persistence infection in hotspot villages. “Hotspots villages” were defined in our study as those reporting more than six indigenous malaria cases during the previous year. A total of ten villages, including seven hotspots and three non-hotspots, were surveyed. All potential mosquito breeding sites identified in and around the ten study villages were regularly monitored between 2013 and 2017. Monitoring comprised the detection of anopheline larvae and the collection of epidemiological, hydrogeological, topographical, and biogeographical data. The number of larval breeding sites described and monitored during the study period ranged from 50 to 62. Breeding sites were more numerous in hotspot sites in each year of monitoring, with 90.3% (56/62) in 2013, 90.9% (50/55) in 2014, 90.3% (56/62) in 2015 and 86% (43/50) in 2017 (Fisher exact test; p = 1). In the non-hotspot areas, the data for the same years were, respectively, 9.7% (6/62), 9.1% (5/55), 9.7% (6/62) and 14% (7/50) (p = 1). The Hotspot villages were characterized mostly by saline or moderately saline hydro-morphic and halomorphic soils allowing water retention and a potential larval breeding sites. By contrast, non-hotspot villages were characterized mainly by a high proportion of extremely permeable sandy-textured soils, which due to their porosity had low water retention. The annual number of confirmed malaria cases was correlated with the frequency and extent of breeding sites. Malaria cases were significantly more frequent in the hamlets located near breeding sites of An. gambiae s.l., gradually decreasing with increasing remoteness. This study shows that the characteristics of larval breeding sites, as measured by their longevity, stability, proximity to human habitation, and their positivity in Anopheles larvae are likely determining factors in the persistence of malaria hotspots in central-western Senegal. The results of this study shed more light on the environmental factors underlying the residual transmission and should make it possible to better target vector control interventions for malaria elimination in west-central Senegal.

Environmental Factors Associated with Larval Habitats of Anopheline Mosquitoes (Diptera: Culicidae) in Metema District, Northwestern Ethiopia

Background: Malaria is one of the major public health concerns in Ethiopia. Control options available for the management of malaria, include case detection, personal protection and larval source management. Effective control of Anopheles larvae largely depends on understanding of the habitats of the vectors. The aims of this study were to identify the breeding habitats of mosquitoes and characterize the larval habitats in Gende Wuha Town in northwestern Ethiopia. Methods: Different aquatic habitats were sampled and characterized for anopheline larvae from November 2012 to June 2013 Results: In total, 2784 larvae of Anopheles mosquitoes were collected from various breeding habitats. Microscopic identification of the III and IV instars revealed the presence of seven Anopheles species. Of the Anopheles spp, Anopheles gambiae s.l. (80%) was the most predominant species in the study area. Spearman correlation coefficient results also determined that the density of An. gambiae s.l. increased significantly with habitat temperature (r= 0.346, p< 0.01). Significantly higher An. gambiae s.l. larvae were obtained in non-shaded habitats (z= -3.120, p< 0.05) when compared with shaded habitats. Conclusions: The current study demonstrated An. gambiae s.l., the principal malaria vector in the country, is the predominant species in the larval sampling habitats. It was also noted the importance of edge of stream as larva breeding habitats for this species during the dry season of the year. Therefore, attention should be given for this breeding habitat for control of the vector during dry season.

Spatial distribution of Anopheles gambiae sensu lato larvae in the urban environment of Yaoundé, Cameroon

Background The rapid and unplanned urbanization of African cities is considered to increase the risk of urban malaria transmission. The present study objective was to assess factors influencing the spatio-temporal distribution of Anopheles gambiae s.l. larvae in the city of Yaoundé, Cameroon. Methods All water bodies were checked once every 2 months for the presence of mosquito larvae from March 2017 to May 2018 in 32 districts of Yaoundé. Physico-chemical characteristics including the size, depth, turbidity, pH, temperature, conductivity, sulfates, organophosphates, hydrogen peroxide (H2O2), conductivity, iron and calcium were recorded and analyzed according to anopheline larvae presence or absence. High resolution satellite images from landsat sentinel Enhanced Thematic Mapper were used for spatial mapping of both field and environmental variables. Bivariate and multivariate logistic regression models were used to identify variables closely associated with anopheline larvae distribution. Results A total of 18 696 aquatic habitats were checked and only 2942 sites (15.7%) contained anopheline larvae. A high number of sites with anopheline larvae (≥ 69%) presented late instar larvae (L3, L4 and pupae). Anopheline mosquito larvae were sampled from a variety of breeding sites including puddles (51.6%), tire prints (12.9%), wells (11.7%) and drains (11.3%). Bivariate logistic regression analyses associated anopheline larvae presence with the absence of predators, absence of algae, absence of vegetation and depth of less than 1 m. Conductivity, turbidity, organophosphates, H2O2 and temperature were significantly high in breeding sites with anopheline larvae than in breeding sites without these larvae (P < 0.1). Anopheline species collected included An. coluzzii (91.1%) and An. gambiae s.s. (8.9%). GIS mapping indicated a heterogeneous distribution of anopheline breeding habitats in the city of Yaoundé. Land cover analysis indicated high variability of the city of Yaoundé’s landscape. Conclusions The data confirms adaptation of An. gambiae s.l. to the urban domain in the city of Yaoundé and calls for urgent actions to improve malaria vector control.

Isolation ofBacillus sphaericusfrom Lombok Island, Indonesia, and Their Toxicity againstAnopheles aconitus

Malaria is endemic to Lombok Island, Indonesia. One approach to suppress malaria spread is to eliminate anopheline larvae in their habitat and the environmentally safe agent is bacteria, that is,Bacillus sphaericus. However, there is no information regarding local isolate ofB. sphaericusthat is toxic to mosquito larvae from Lombok. The aims of the study were to isolateB. sphaericusfrom soil in areas close to beach surrounding Lombok Island and to test their toxicity against 3rd instarAnopheles aconituslarvae. Soil samples were collected from 20 different sampling locations from Lombok Island and homogenized with sterile physiological salt solution. Suspension was heat-shocked at 80°C for 30 minutes and then spread onto antibiotic-supplemented NYSM solid medium. Colonies grown were characterized and subjected to initial toxicity test against anopheline larvae. Isolates with more than 50% killing percentage were subjected to bioassay testing against anopheline larvae. From 20 locations, 1 isolate showed mild toxicity (namely, isolate MNT) and 2 isolates showed high toxicity (namely, isolates SLG and TJL2) againstAn. aconitus. Those 3 isolates were potentially useful isolates, as they killed almost all larvae in 24 hours. The discovery of toxic indigenous isolates ofB. sphaericusfrom Lombok Island opens opportunity to develop a biopesticide from local resources.