Evaluation of the DN-Mini (miniaturized double net) trap for sampling host-seeking Anopheles mosquitoes in malaria-endemic villages of southern Tanzania.

BACKGROUND: Surveillance of malaria vectors is crucial for assessing the transmission risk and impact of control measures. Human landing catches (HLC) directly estimate the biting rates but raise ethical concerns due to the exposure of volunteers to mosquito-borne pathogens. A common alternative is the CDC-light trap, which is effective for catching host-seeking mosquitoes indoors but not outdoors. New, exposure-free methods are needed for sampling mosquitoes indoors and outdoors in ways that reflect their natural risk profiles. The aim of this study was therefore to evaluate the efficacy of the miniaturized double net trap (DN-Mini) for sampling host-seeking mosquitoes in south-eastern Tanzania, where malaria transmission is dominated by Anopheles funestus. METHODS: Adult mosquitoes were collected from 222 randomly selected houses across three villages (74 per village) in Ulanga district, south-eastern Tanzania, using the DN-Mini traps, CDC-Light traps, and Prokopack aspirators. First, we compared CDC-light and DN-Mini traps for collecting indoor host-seeking mosquitoes, while Prokopack aspirators were used for indoor-resting mosquitoes. Second, we deployed the DN-Mini and Prokopack aspirators to collect host-seeking and resting mosquitoes indoors and outdoors. Generalized linear mixed models (GLMM) with a negative binomial distribution were used to compare the effectiveness of the traps for catching different mosquito species. RESULTS: The DN-Mini was 1.53 times more efficient in collecting An. funestus indoors (RR = 1.53, 95% CI: 1.190-1.98) compared to the CDC-Light trap. However, for Anopheles arabiensis, the DN-Mini caught only 0.32 times as many mosquitoes indoors as the CDC-Light traps (RR = 0.32, 95% CI: 0.183-0.567). Both An. funestus and An. arabiensis were found to be more abundant indoors than outdoors when collected using the DN-Mini trap. Similarly, the Prokopack aspirator was greater indoors than outdoors for both An. funestus and An. arabiensis. CONCLUSION: The DN-Mini outperformed the CDC-light trap in sampling the dominant malaria vector, An. funestus species, but was less effective in capturing An. arabiensis, and for both vector species, the biting risk was greater indoors than outdoors when measured using the DN-Mini trap. These findings highlight the importance of selecting appropriate trapping methods based on mosquito species and behaviors.

Corrigendum to "Persistently high proportions of plasmodium-infected Anopheles funestus mosquitoes in two villages in the Kilombero valley, South-Eastern Tanzania" [Parasite Epidemiology and Control 18 (2022) e00264].

[This corrects the article DOI: 10.1016/j.parepi.2022.e00264.].

Persistently high proportions of plasmodium-infected Anopheles funestus mosquitoes in two villages in the Kilombero valley, South-Eastern Tanzania.

Background: In south-eastern Tanzania where insecticide-treated nets have been widely used for >20 years, malaria transmission has greatly reduced but remains highly heterogenous over small distances. This study investigated the seasonal prevalence of Plasmodium sporozoite infections in the two main malaria vector species, Anopheles funestus and Anopheles arabiensis for 34 months, starting January 2018 to November 2020. Methods: Adult mosquitoes were collected using CDC-light traps and Prokopack aspirators inside local houses in Igumbiro and Sululu villages, where earlier surveys had found very high densities of An. funestus. Collected females were sorted by taxa, and the samples examined using ELISA assays for detecting Plasmodium circumsporozoite protein in their salivary glands. Results: Of 7859 An. funestus tested, 4.6% (n = 365) were positive for Pf sporozoites in the salivary glands. On the contrary, only 0.4% (n = 9) of the 2382 An. arabiensis tested were positive. The sporozoite prevalence did not vary significantly between the villages or seasons. Similarly, the proportions of parous females of either species were not significantly different between the two villages (p > 0.05) but was slightly higher in An. funestus (0.50) than in An. arabiensis (0.42). Analysis of the 2020 data determined that An. funestus contributed 97.7% of all malaria transmitted in households in these two villages. Conclusions: In contexts where individual vector species mediate most of the pathogen transmission, it may be most appropriate to pursue a species-focused approach to better understand the ecology of the dominant vectors and target them with effective interventions to suppress transmission. Despite the ongoing efforts on tackling malaria in the two study villages, there is still persistently high Plasmodium infection prevalence in local populations of An. funestus, which now carry ~97% of all malaria infections and mediates intense year-round transmission. Further reduction in malaria burden in these or other similar settings requires effective targeting of An. funestus.

Fine-scale distribution of malaria mosquitoes biting or resting outside human dwellings in three low-altitude Tanzanian villages.

BACKGROUND: While malaria transmission in Africa still happens primarily inside houses, there is a substantial proportion of Anopheles mosquitoes that bite or rest outdoors. This situation may compromise the performance of indoor insecticidal interventions such as insecticide-treated nets (ITNs). This study investigated the distribution of malaria mosquitoes biting or resting outside dwellings in three low-altitude villages in south-eastern Tanzania. The likelihood of malaria infections outdoors was also assessed. METHODS: Nightly trapping was done outdoors for 12 months to collect resting mosquitoes (using resting bucket traps) and host-seeking mosquitoes (using odour-baited Suna® traps). The mosquitoes were sorted by species and physiological states. Pooled samples of Anopheles were tested to estimate proportions infected with Plasmodium falciparum parasites, estimate proportions carrying human blood as opposed to other vertebrate blood and identify sibling species in the Anopheles gambiae complex and An. funestus group. Environmental and anthropogenic factors were observed and recorded within 100 meters from each trapping positions. Generalised additive models were used to investigate relationships between these variables and vector densities, produce predictive maps of expected abundance and compare outcomes within and between villages. RESULTS: A high degree of fine-scale heterogeneity in Anopheles densities was observed between and within villages. Water bodies covered with vegetation were associated with 22% higher densities of An. arabiensis and 51% lower densities of An. funestus. Increasing densities of houses and people outdoors were both associated with reduced densities of An. arabiensis and An. funestus. Vector densities were highest around the end of the rainy season and beginning of the dry seasons. More than half (14) 58.3% of blood-fed An. arabiensis had bovine blood, (6) 25% had human blood. None of the Anopheles mosquitoes caught outdoors was found infected with malaria parasites. CONCLUSION: Outdoor densities of both host-seeking and resting Anopheles mosquitoes had significant heterogeneities between and within villages, and were influenced by multiple environmental and anthropogenic factors. Despite the high Anopheles densities outside dwellings, the substantial proportion of non-human blood-meals and absence of malaria-infected mosquitoes after 12 months of nightly trapping suggests very low-levels of outdoor malaria transmission in these villages.

Evaluation of a push-pull system consisting of transfluthrin-treated eave ribbons and odour-baited traps for control of indoor- and outdoor-biting malaria vectors.

BACKGROUND: Push-pull strategies have been proposed as options to complement primary malaria prevention tools, indoor residual spraying (IRS) and long-lasting insecticide-treated nets (LLINs), by targeting particularly early-night biting and outdoor-biting mosquitoes. This study evaluated different configurations of a push-pull system consisting of spatial repellents [transfluthrin-treated eave ribbons (0.25 g/m2 ai)] and odour-baited traps (CO2-baited BG-Malaria traps), against indoor-biting and outdoor-biting malaria vectors inside large semi-field systems. METHODS: Two experimental huts were used to evaluate protective efficacy of the spatial repellents (push-only), traps (pull-only) or their combinations (push-pull), relative to controls. Adult volunteers sat outdoors (1830 h-2200 h) catching mosquitoes attempting to bite them (outdoor-biting risk), and then went indoors (2200 h-0630 h) to sleep under bed nets beside which CDC-light traps caught host-seeking mosquitoes (indoor-biting risk). Number of traps and their distance from huts were varied to optimize protection, and 500 laboratory-reared Anopheles arabiensis released nightly inside the semi-field chambers over 122 experimentation nights. RESULTS: Push-pull offered higher protection than traps alone against indoor-biting (83.4% vs. 35.0%) and outdoor-biting (79% vs. 31%), but its advantage over repellents alone was non-existent against indoor-biting (83.4% vs. 81%) and modest for outdoor-biting (79% vs. 63%). Using two traps (1 per hut) offered higher protection than either one trap (0.5 per hut) or four traps (2 per hut). Compared to original distance (5 m from huts), efficacy of push-pull against indoor-biting peaked when traps were 15 m away, while efficacy against outdoor-biting peaked when traps were 30 m away. CONCLUSION: The best configuration of push-pull comprised transfluthrin-treated eave ribbons plus two traps, each at least 15 m from huts. Efficacy of push-pull was mainly due to the spatial repellent component. Adding odour-baited traps slightly improved personal protection indoors, but excessive trap densities increased exposure near users outdoors. Given the marginal efficacy gains over spatial repellents alone and complexity of push-pull, it may be prudent to promote just spatial repellents alongside existing interventions, e.g. LLINs or non-pyrethroid IRS. However, since both transfluthrin and traps also kill mosquitoes, and because transfluthrin can inhibit blood-feeding, field studies should be done to assess potential community-level benefits that push-pull or its components may offer to users and non-users.

Eave ribbons treated with the spatial repellent, transfluthrin, can effectively protect against indoor-biting and outdoor-biting malaria mosquitoes.

BACKGROUND: Long-lasting insecticide-treated nets and indoor residual spraying protect against indoor-biting and indoor-resting mosquitoes but are largely ineffective for early-biting and outdoor-biting malaria vectors. Complementary tools are, therefore, needed to accelerate control efforts. This paper describes simple hessian ribbons treated with spatial repellents and wrapped around eaves of houses to prevent outdoor-biting and indoor-biting mosquitoes over long periods of time. METHODS: The eave ribbons are 15 cm-wide triple-layered hessian fabrics, in lengths starting 1 m. They can be fitted onto houses using nails, adhesives or Velcro, without completely closing eave-spaces. In 75 experimental nights, untreated ribbons and ribbons treated with 0.02%, 0.2%, 1.5% or 5% transfluthrin emulsion (spatial repellent) were evaluated against blank controls using two experimental huts inside a 202 m2 semi-field chamber where 500 laboratory-reared Anopheles arabiensis were released nightly. Two volunteers sat outdoors (one/hut) and collected mosquitoes attempting to bite them from 6 p.m. to 10 p.m. (outdoor-biting), then went indoors and slept under bed nets, beside which CDC-light traps collected mosquitoes from 10 p.m. to 6.30 a.m. (indoor-biting). To assess survival, 200 caged mosquitoes were suspended near the huts nightly and monitored for 24 h thereafter. Additionally, field tests were done in experimental huts in a rural Tanzanian village to evaluate treated ribbons (1.5% transfluthrin). Here, indoor-biting was assessed using window traps and Prokopack® aspirators, and outdoor-biting assessed using volunteer-occupied double-net traps. RESULTS: Indoor-biting and outdoor-biting decreased > 99% in huts fitted with eave ribbons having ≥ 0.2% transfluthrin. Even 0.02% transfluthrin-treated ribbons provided 79% protection indoors and 60% outdoors. Untreated ribbons however reduced indoor-biting by only 27% and increased outdoor-biting by 18%, though these were non-significant (P > 0.05). Of all caged mosquitoes exposed near treated huts, 99.5% died within 24 h. In field tests, the ribbons provided 96% protection indoors and 84% outdoors against An. arabiensis, plus 42% protection indoors and 40% outdoors against Anopheles funestus. Current prototypes cost ~ 7USD/hut, are made of widely-available hessian and require no specialized expertise. CONCLUSION: Transfluthrin-treated eave ribbons significantly prevented outdoor-biting and indoor-biting malaria vectors and could potentially complement current tools. The technique is simple, low-cost, highly-scalable and easy-to-use; making it suitable even for poorly-constructed houses and low-income groups.

Small-scale field evaluation of push-pull system against early- and outdoor-biting malaria mosquitoes in an area of high pyrethroid resistance in Tanzania.

Background: Despite high coverage of indoor interventions like insecticide-treated nets, mosquito-borne infections persist, partly because of outdoor-biting, early-biting and insecticide-resistant vectors. Push-pull systems, where mosquitoes are repelled from humans and attracted to nearby lethal targets, may constitute effective complementary interventions. Methods: A partially randomized cross-over design was used to test efficacy of push-pull in four experimental huts and four local houses, in an area with high pyrethroid resistance in Tanzania. The push-pull system consisted of 1.1% or 2.2% w/v transfluthrin repellent dispensers and an outdoor lure-and-kill device (odour-baited mosquito landing box). Matching controls were set up without push-pull. Adult male volunteers collected mosquitoes attempting to bite them outdoors, but collections were also done indoors using exit traps in experimental huts and by volunteers in the local houses. The collections were done hourly (1830hrs-0730hrs) and mosquito catches compared between push-pull and controls. An. gambiae s.l. and An. funestus s.l. were assessed by PCR to identify sibling species, and ELISA to detect Plasmodium falciparum and blood meal sources. Results: Push-pull in experimental huts reduced outdoor-biting for An. arabiensis and Mansonia species by 30% and 41.5% respectively. However, the reductions were marginal and insignificant for An. funestus (12.2%; p>0.05) and Culex (5%; p>0.05). Highest protection against all species occurred before 2200hrs. There was no significant difference in number of mosquitoes inside exit traps in huts with or without push-pull. In local households, push-pull significantly reduced indoor and outdoor-biting of An. arabiensis by 48% and 25% respectively, but had no effect on other species. Conclusion: This push-pull system offered modest protection against outdoor-biting An. arabiensis, without increasing indoor mosquito densities. Additional experimentation is required to assess how transfluthrin-based products affect mosquito blood-feeding and mortality in push-pull contexts. This approach, if optimised, could potentially complement existing malaria interventions even in areas with high pyrethroid resistance.