Analysis of the 24-h biting patterns and human exposures to malaria vectors in south-eastern Tanzania.

BACKGROUND: Afrotropical malaria vectors are generally believed to bite nocturnally, leading to the predominant use of insecticide-treated nets (ITNs), which target indoor, nighttime-biting mosquitoes. This focus is reinforced by biases in entomological surveys, which largely overlook daytime mosquito activity. However, recent evidence challenges this paradigm, showing that Anopheles biting can extend way into the daytime, coinciding with human activities at dawn, daytime and evenings, suggesting a broader risk spectrum and potential protection gaps. We have therefore investigated the diurnal and nocturnal biting patterns of the malaria vectors Anopheles arabiensis and Anopheles funestus in south-eastern Tanzania, to better understand the scope of residual transmission and inform strategies for improved control. METHODS: Host-seeking mosquitoes were collected hourly using miniaturized double net traps, both indoors and outdoors over 24-h periods between June 2023 and February 2024. Concurrently, human activities indoors and outdoors were monitored half-hourly to correlate with mosquito collections. A structured questionnaire was used to assess household members' knowledge, perceptions and experiences regarding exposure to mosquito bites during both nighttime and daytime. RESULTS: Nocturnal biting by An. arabiensis peaked between 7 p.m. and 11 p.m. while that of An. funestus peaked later, between 1 a.m. and 3 a.m. Daytime biting accounted for 15.03% of An. arabiensis catches, with peaks around 7-11 a.m. and after 4 p.m., and for 14.15% of An. funestus catches, peaking around mid-mornings, from 10 a.m. to 12 p.m. Nighttime exposure to An. arabiensis was greater outdoors (54.5%), while daytime exposure was greater indoors (80.4%). For An. funestus, higher exposure was observed indoors, both at nighttime (57.1%) and daytime (69%). Plasmodium falciparum sporozoites were detected in both day-biting and night-biting An. arabiensis. Common daytime activities potentially exposing residents during peak biting hours included household chores, eating, sleeping (including due to sickness), resting in the shade or under verandas and playing (children). From evenings onwards, exposures coincided with resting, socializing before bedtime and playtime (children). Nearly all survey respondents (95.6%) reported experiencing daytime mosquito bites, but only 28% believed malaria was transmissible diurnally. CONCLUSIONS: This study updates our understanding of malaria vector biting patterns in south-eastern Tanzania, revealing considerable additional risk in the mornings, daytime and evenings. Consequently, there may be more gaps in the protection provided by ITNs, which primarily target nocturnal mosquitoes, than previously thought. Complementary strategies are needed to holistically suppress vectors regardless of biting patterns (e.g. using larval source management) and to extend personal protection limits (e.g. using repellents). Additionally, community engagement and education on mosquito activity and protective measures could significantly reduce malaria transmission risk.

Semi-field evaluation of aquatic predators for the control of Anopheles funestus in rural south-eastern Tanzania.

BACKGROUND: Biological control is a promising alternative or complementary approach for controlling vector populations in response to the spread of insecticide resistance in malaria vectors. This study evaluated the efficacy of three selected potential predators on the density and fitness parameters of Anopheles funestus larvae in rural Tanzania. METHODS: Common predator families Aeshnidae (dragonflies), Coenagrionidae (damselflies), and Notonectidae (backswimmers) and An. funestus group larvae were collected from natural aquatic habitats in rural south-eastern Tanzania. Predators were starved for 12-h while An. funestus larvae were given fish food before starting the experiment. Anopheles funestus larvae were placed into artificial habitats containing predators, exposing them to potential predation. The number of surviving An. funestus larvae were counted every 24-h. An emergence traps were placed at the top of artificial habitats to capture emerging mosquitoes. Emerged mosquitoes were monitored until they died. Female wings were measured and used as a proxy for body size. Generalized linear mixed models (GLMM) with binomial variates at 95% CI and Cox proportional hazard models were used to assess the proportion of dead mosquitoes and the daily survival determined. RESULTS: There were significant differences in the number of emerged mosquitoes between the treatment and control groups (P < 0.001). Thus, all predator species played a significant role in reducing the density of An. funestus mosquitoes (P < 0.001). Furthermore, these predators had notable effects on the fitness parameters and survival of emerged mosquitoes (P < 0.001). Among the three predators studied, Coenagrionidae (damselflies) were most efficient followed by Notonectidae (backswimmers), with Aeshnidae (dragonflies) being the least efficient. CONCLUSION: Selected aquatic predators have the potential to reduce the survival and density of An. funestus larvae. They might eventually be included within an integrated malaria vector control strategy, ultimately leading to a reduction in malaria transmission.

Livestock keeping, mosquitoes and community viewpoints: a mixed methods assessment of relationships between livestock management, malaria vector biting risk and community perspectives in rural Tanzania.

BACKGROUND: Livestock keeping is one of the potential factors related to malaria transmission. To date, the impact of livestock keeping on malaria transmission remains inconclusive, as some studies suggest a zooprophylactic effect while others indicate a zoopotentiation effect. This study assessed the impact of livestock management on malaria transmission risks in rural Tanzania. Additionally, the study explored the knowledge and perceptions of residents about the relationships between livestock keeping and malaria transmission risks in a selected village. METHODS: In a longitudinal entomological study in Minepa village, South Eastern Tanzania, 40 households were randomly selected (20 with livestock, 20 without). Weekly mosquito collection was performed from January to April 2023. Indoor and outdoor collections used CDC-Light traps, Prokopack aspirators, human-baited double-net traps, and resting buckets. A subsample of mosquitoes was analysed using PCR and ELISA for mosquito species identification and blood meal detection. Livestock's impact on mosquito density was assessed using negative binomial GLMMs. Additionally, in-depth interviews explored community knowledge and perceptions of the relationship between livestock keeping and malaria transmission risks. RESULTS: A total of 48,677 female Anopheles mosquitoes were collected. Out of these, 89% were Anopheles gambiae sensu lato (s.l.) while other species were Anopheles funestus s.l., Anopheles pharoensis, Anopheles coustani, and Anopheles squamosus. The findings revealed a statistically significant increase in the overall number of An. gambiae s.l. outdoors (RR = 1.181, 95%CI 1.050-1.862, p = 0.043). Also, there was an increase of the mean number of An. funestus s.l. mosquitoes collected in households with livestock indoors (RR = 2.866, 95%CI: 1.471-5.582, p = 0.002) and outdoors (RR = 1.579,95%CI 1.080-2.865, p = 0.023). The human blood index of Anopheles arabiensis mosquitoes from houses with livestock was less than those without livestock (OR = 0.149, 95%CI 0.110-0.178, p < 0.001). The majority of participants in the in-depth interviews reported a perceived high density of mosquitoes in houses with livestock compared to houses without livestock. CONCLUSION: Despite the potential for zooprophylaxis, this study indicates a higher malaria transmission risk in livestock-keeping communities. It is crucial to prioritize and implement targeted interventions to control vector populations within these communities. Furthermore, it is important to enhance community education and awareness regarding covariates such as livestock that influence malaria transmission.

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.

Challenges of proper disposal of old long-lasting insecticidal nets and its alternative uses in rural south-eastern Tanzania.

INTRODUCTION: Insecticide-treated nets (ITNs), specifically long-lasting insecticidal nets (LLINs), are the most commonly used, scalable, and cost-effective tools for controlling malaria transmission in sub-Saharan Africa. However, the multiple alternative uses of retired LLINs have been associated with poor disposal practices. The World Health Organization (WHO) has provided guidelines and recommendations for the proper management of worn-out LLINs. This study assessed the existing alternative uses and disposal practices of old LLINs. METHODS: An explanatory sequential mixed-methods approach was used to assess LLINs existing alternative uses, disposal practices, knowledge, and perceptions regarding WHO recommendations on proper disposal of old LLINs among stakeholders in Kilombero and Ulanga districts, south-eastern Tanzania. A survey questionnaire was administered to 384 participants. Furthermore, the study employed focus group discussions (FGD) and key informant interviews (KII) to elucidate responses regarding existing disposal practices, associated challenges, and alternative uses of LLINs. The insights derived from both study components were subsequently used for inferential analysis. RESULTS: The major challenge influencing the proper disposal of LLINs was limited awareness of how to properly dispose of them. Of the 384 people surveyed, 97.0% were not aware of the WHO recommendations for the proper disposal of old LLINs. All key informants were unaware of the WHO guidelines for proper disposal of old LLINs. The common methods used to dispose of LLINs were burning (30.7%), disposing them into garbage pits (14.8%), and alternative uses (12.2%). Of the 239 respondents with LLINs, 41.0% had alternative use, while 59.0% had no alternative use. The common alternative uses were ropes for tying or covering items (20.9%), garden fencing (7.5%), chicken coops (5.0%), and 7.5% for other minor alternative uses. CONCLUSION: Strengthening awareness and education on proper LLIN disposal practices among community members and key stakeholders is essential for enhancing malaria control efforts and preventing environmental pollution.

Sterilized Anopheles funestus can autodisseminate sufficient pyriproxyfen to the breeding habitat under semi-field settings.

BACKGROUND: Anopheles funestus, the main malaria vector, prefer to oviposit in permanent and/or semi-permanent breeding habitats located far from human dwellings. Difficulties in identifying and accessing these habitats jeopardize the feasibility of conventional larviciding. In this way, a semi-field study was conducted to assess the potential of autodissemination of pyriproxyfen (PPF) by An. funestus for its control. METHODS: The study was conducted inside a semi-field system (SFS). Therein, two identical separate chambers, the treatment chamber with a PPF-treated clay pot (0.25 g AI), and the control chamber with an untreated clay pot. In both chambers, one artificial breeding habitat made of a plastic basin with one litre of water was provided. Three hundred blood-fed female An. funestus aged 5-9 days were held inside untreated and treated clay pots for 30 min and 48 h before being released for oviposition. The impact of PPF on adult emergence, fecundity, and fertility through autodissemination and sterilization effects were assessed by comparing the treatment with its appropriate control group. RESULTS: Mean (95% CI) percentage of adult emergence was 15.5% (14.9-16.1%) and 70.3% (69-71%) in the PPF and control chamber for females exposed for 30 min (p < 0.001); and 19% (12-28%) and 95% (88-98%) in the PPF and control chamber for females exposed for 48 h (p < 0.001) respectively. Eggs laid by exposed mosquitoes and their hatch rate were significantly reduced compared to unexposed mosquitoes (p < 0.001). Approximately, 90% of females exposed for 48 h retained abnormal ovarian follicles and only 42% in females exposed for 30 min. CONCLUSION: The study demonstrated sterilization and adult emergence inhibition via autodissemination of PPF by An. funestus. Also, it offers proof that sterilized An. funestus can transfer PPF to prevent adult emergence at breeding habitats. These findings warrant further assessment of the autodissemination of PPF in controlling wild population of An. funestus, and highlights its potential for complementing long-lasting insecticidal nets.