Larval ecology of malaria vectors and the impact of larviciding on malaria transmission in The Gambia

The study reported in this thesis explored the ecology of aquatic stages of mosquitoes in the middle reaches of the Gambia River in order to assess the feasibility and impact of microbial larviciding on malaria transmission in large river ecosystems in sub- Saharan Africa. All accessible water bodies in four study zones covering 400 km(^2) were mapped and sampled for mosquitoes. Microbial larvicides were applied in the four zones in across-over design and the impact of larviciding on mosquito densities assessed. Anopheline and culicine mosquitoes were found in all sampled habitats, apart from those with moving water. Similarly, all habitats, except puddles and water channels, had similar larval and pupal densities. Anopheles gambiae sensu lato, the major malaria vector in Africa, exploited a wide range of habitats and despite a decrease in population density during the dry season, could be found in breeding sites throughout the year. Mosquitoes shared habitats with other invertebrates including their predators. A closer look at rice fields revealed that mosquitoes were abundant in rice fields closer to the landward edge of the floodplains where water is fresher and contains high quantifies of nutrients. Mosquitoes of The Gambia were highly susceptible to both Bacillus thuringiensis var. israelensis (Bti) and B. sphaericus microbials, however no residual activity against anopheline larvae was observed. The basic training of personnel in identification of habitats, calibration of application equipment and active larviciding proved to be successful. Routine larviciding was associated with > 91 % reducfion (p < 0.001) in anophelines late stage larval density and 72 % (p < 0.001) in culicines. Overall, larviciding was associated with a 28% (p = 0.005) reduction in the number of adult female Anopheles gambiae s.l. found indoors, although this rose to 42%, when the study zone with the greatest abundance of breeding sites was excluded from the analysis. No significant reduction in adult culicines was observed. Ground application of Bti in areas with extensive floodplains is unlikely to contribute to a substantial reduction in malaria transmission in The Gambia, therefore vector control in such areas should target adult mosquitoes

A Simple and Efficient Tool for Trapping Gravid Anopheles at Breeding Sites.

No effective tool currently exists for trapping ovipositing malaria vectors. This creates a gap in our ability to investigate the behavior and ecology of gravid Anopheles.\ud Here we describe a simple trap that collects ovipositing Anopheline and Culicine mosquitoes. It consists of an acetate sheet coated in glue that floats on the water surface. Ten breeding sites were selected in rural Tanzania and 10 sticky traps set in each. These caught a total of 74 gravid Anopheles (54 An. arabiensis, 1 An. gambiae s.s. and 16 unamplified) and 1333 gravid Culicines, in just two trap nights. This simple sampling tool provides an opportunity to further our understanding of the behavior and ecology of gravid female Anophelines. It strongly implies that at least two of the major vectors of malaria in Africa land on the water surface during the oviposition process, and demonstrates that Anophelines and Culicines often share the same breeding sites. This simple and efficient trap has clear potential for the study of oviposition site choice and productivity, gravid dispersal, and vector control techniques which use oviposition behavior as a means of disseminating larvicides

Biologically meaningful coverage indicators for eliminating malaria transmission.

Mosquitoes, which evade contact with long-lasting insecticidal nets and indoor residual sprays, by feeding outdoors or upon animals, are primary malaria vectors in many tropical countries. They can also dominate residual transmission where high coverage of these front-line vector control measures is achieved. Complementary strategies, which extend insecticide coverage beyond houses and humans, are required to eliminate malaria transmission in most settings. The overwhelming diversity of the world's malaria transmission systems and optimal strategies for controlling them can be simply conceptualized and mapped across two-dimensional scenario space defined by the proportion of blood meals that vectors obtain from humans and the proportion of human exposure to them which occurs indoors

Larviciding to prevent malaria transmission

Background Larviciding refers to the regular application of chemical or microbial insecticides to water bodies or water containers to kill the aquatic immature forms of the mosquito (the larvae and pupae). Objectives To summarize research evidence evaluating whether larviciding with chemical or microbial insecticides prevents malaria transmission. Search methods We searched the Cochrane Infectious Diseases Group Specialized Register; the Cochrane Central Register of Controlled Trials (CENTRAL), published in the Cochrane Library; MEDLINE; Embase; CAB Abstracts; LILACS; the World Health Organization International Clinical Trials Registry Platform (WHO ICTRP); ClinicalTrials.gov; and the ISRCTN registry up to 6 June 2019. Selection criteria We included cluster‐randomized controlled trials (cRCTs), interrupted time series (ITS), randomized cross‐over studies, non‐randomized cross‐over studies, and controlled before‐and‐after studies (CBAs) that compared larviciding with no larviciding. Data collection and analysis We independently assessed trials for eligibility and risk of bias, and extracted data. We assessed the certainty of evidence using the GRADE approach. Main results Four studies (one cRCT, two CBAs, and one non‐randomized cross‐over design) met the inclusion criteria. All used ground application of larvicides (people hand‐delivering larvicides); one evaluated chemical and three evaluated microbial agents. Studies were carried out in The Gambia, Tanzania, Kenya, and Sri Lanka. Three studies were conducted in areas where mosquito aquatic habitats were less extensive ( 1 km²; a cross‐over study from The Gambia). For aquatic habitats of less than 1 km², one cRCT randomized eight villages in Sri Lanka to evaluate chemical larviciding using insect growth regulator; and two CBA studies undertaken in Kenya and Tanzania evaluated microbial larvicides. In the cRCT, larviciding across all villages was associated with lower malaria incidence (rate ratio 0.24, 4649 participants, low‐certainty evidence) and parasite prevalence (risk ratio (RR) 0.26, 5897 participants, low‐certainty evidence) compared to no larviciding. The two CBA studies reported lower malaria prevalence during the intervention period (parasite prevalence RR 0.79, 95% confidence interval (CI) 0.71 to 0.89; 70,902 participants; low‐certainty evidence). The Kenyan study also reported a reduction in the incidence of new malaria cases (RR 0.62, 95% CI 0.38 to 1.01; 720 participants; very low‐certainty evidence). For aquatic habitats of more than 1 km², the non‐randomized cross‐over trial using microbial larvicides did not detect an effect for malaria incidence (RR 1.58, 95% CI 0.94 to 2.65; 4226 participants), or parasite prevalence (RR 1.15, 95% CI 0.41 to 3.20; 3547 participants); both were very low‐certainty evidence. The Gambia trial also reported the mean haemoglobin level, and there was no difference across the four comparisons (mean difference –0.13, 95% CI –0.40 to 0.13; 3586 participants). We were unable to summarize or pool entomological outcomes due to unreported and missing data. Authors' conclusions Most controlled studies on larviciding have been performed with microbial agents. Ground larviciding for non‐extensive larval habitats may have an effect on malaria transmission, and we do not know if there is an effect in large‐scale aquatic habitats. We found no studies using larviciding application techniques that could cover large aquatic habitats, such as aerial spraying using aircraft

'Like sugar and honey': the embedded ethics of a larval control project in The Gambia.

This paper describes a malaria research project in The Gambia to provoke thinking on the social value of transnational research. The Larval Control Project (LCP) investigated the efficacy of a microbial insecticide to reduce vector density and, ultimately, clinical malaria in Gambian children. The LCP's protocol delineated a clinical surveillance scheme that involved Village Health Workers (VHWs) supported by project nurses. Combining insights from ethnographic fieldwork conducted at the Medical Research Council (MRC) Laboratories in Farafenni from 2005 to 2009, open-ended interviews with project nurses, and eight focus group discussions held with participant mothers in October 2007, we consider the social impact of the LCP's investigative method against the backdrop of several years of research activity. We found that while participants associated the LCP with the clinical care it provided, they also regarded the collaboration between the nurses and VHWs added additional benefits. Organised around the operational functions of the trial, small-scale collaborations provided the platform from which to build local capacity. While ethical guidelines emphasise the considerations that must be added to experimental endeavour in southern countries (e.g. elaborating processes of informed consent, developing strategies of community engagement or providing therapeutic access to participants after the trial concludes), these findings suggest that shifting attention from supplementing ethical protocols to the everyday work of research -embedding ethics through scientific activity - may provide a sounder basis to reinforce the relationship between scientific rigour and social value

Challenges for Malaria Elimination in Zanzibar: Pyrethroid Resistance in Malaria Vectors and Poor Performance of Long-Lasting Insecticide Nets.

Long-lasting insecticide treated nets (LLINs) and indoor residual house spraying (IRS) are the main interventions for the control of malaria vectors in Zanzibar. The aim of the present study was to assess the susceptibility status of malaria vectors against the insecticides used for LLINs and IRS and to determine the durability and efficacy of LLINs on the island. Mosquitoes were sampled from Pemba and Unguja islands in 2010--2011 for use in WHO susceptibility tests. One hundred and fifty LLINs were collected from households on Unguja, their physical state was recorded and then tested for efficacy as well as total insecticide content. Species identification revealed that over 90% of the Anopheles gambiae complex was An. arabiensis with a small number of An. gambiae s.s. and An. merus being present. Susceptibility tests showed that An. arabiensis on Pemba was resistant to the pyrethroids used for LLINs and IRS. Mosquitoes from Unguja Island, however, were fully susceptible to all pyrethroids tested. A physical examination of 150 LLINs showed that two thirds were damaged after only three years in use. All used nets had a significantly lower (p < 0.001) mean permethrin concentration of 791.6 mg/m2 compared with 944.2 mg/m2 for new ones. Their efficacy decreased significantly against both susceptible An. gambiae s.s. colony mosquitoes and wild-type mosquitoes from Pemba after just six washes (p < 0.001). The sustainability of the gains achieved in malaria control in Zanzibar is seriously threatened by the resistance of malaria vectors to pyrethroids and the short-lived efficacy of LLINs. This study has revealed that even in relatively well-resourced and logistically manageable places like Zanzibar, malaria elimination is going to be difficult to achieve with the current control measures

Agriculture and the promotion of insect pests: rice cultivation in river floodplains and malaria vectors in The Gambia

BACKGROUND: Anthropogenic modification of natural habitats can create conditions in which pest species associated with humans can thrive. In order to mitigate for these changes, it is necessary to determine which aspects of human management are associated with the promotion of those pests. Anopheles gambiae, the main Africa malaria vector, often breeds in rice fields. Here the impact of the ancient practice of 'swamp rice' cultivation, on the floodplains of the Gambia River, on the production of anopheline mosquitoes was investigated. METHODS: Routine surveys were carried out along 500 m transects crossing rice fields from the landward edge of the floodplains to the river during the 2006 rainy season. Aquatic invertebrates were sampled using area samplers and emergence traps and fish sampled using nets. Semi-field experiments were used to investigate whether nutrients used for swamp rice cultivation affected mosquito larval abundance. RESULTS: At the beginning of the rainy season rice is grown on the landward edge of the floodplain; the first area to flood with fresh water and one rich in cattle dung. Later, rice plants are transplanted close to the river, the last area to dry out on the floodplain. Nearly all larval and adult stages of malaria vectors were collected 0-100 m from the landward edge of the floodplains, where immature rice plants were grown. These paddies contained stagnant freshwater with high quantities of cattle faeces. Semi-field studies demonstrated that cattle faeces nearly doubled the number of anopheline larvae compared with untreated water. CONCLUSION: Swamp rice cultivation creates ideal breeding sites for malaria vectors. However, only those close to the landward edge harboured vectors. These sites were productive since they were large areas of standing freshwater, rich in nutrients, protected from fish, and situated close to human habitation, where egg-laying mosquitoes from the villages had short distances to fly. The traditional practice of 'swamp rice' cultivation uses different bodies of water on the floodplains to cultivate rice during the rainy season. A consequence of this cultivation is the provizion of ideal conditions for malaria vectors to thrive. As the demand for locally-produced rice grows, increased rice farming will generate great numbers of vectors; emphasizing the need to protect local communities against malaria

Effective autodissemination of pyriproxyfen to breeding sites by the exophilic malaria vector Anopheles arabiensis in semi-field settings in Tanzania

BACKGROUND Malaria vector control strategies that target adult female mosquitoes are challenged by the emergence of insecticide resistance and behavioural resilience. Conventional larviciding is restricted by high operational costs and inadequate knowledge of mosquito-breeding habitats in rural settings that might be overcome by the juvenile hormone analogue, Pyriproxyfen (PPF). This study assessed the potential for Anopheles arabiensis to pick up and transfer lethal doses of PPF from contamination sites to their breeding habitats (i.e. autodissemination of PPF). METHODS A semi-field system (SFS) with four identical separate chambers was used to evaluate PPF-treated clay pots for delivering PPF to resting adult female mosquitoes for subsequent autodissemination to artificial breeding habitats within the chambers. In each chamber, a tethered cow provided blood meals to laboratory-reared, unfed female An. arabiensis released in the SFS. In PPF-treated chambers, clay pot linings were dusted with 0.2 - 0.3 g AI PPF per pot. Pupae were removed from the artificial habitats daily, and emergence rates calculated. Impact of PPF on emergence was determined by comparing treatment with an appropriate control group. RESULTS Mean (95%CI) adult emergence rates were (0.21 +/- 0.299) and (0.95 +/- 0.39) from PPF-treated and controls respectively (p < 0.0001). Laboratory bioassay of water samples from artificial habitats in these experiments resulted in significantly lower emergence rates in treated chambers (0.16 +/- 0.23) compared to controls 0.97 +/- 0.05) (p < 0.0001). In experiments where no mosquitoes introduced, there were no significant differences between control and treatment, indicating that transfer of PPF to breeding sites only occurred when mosquitoes were present; i.e. that autodissemination had occurred. Treatment of a single clay pot reduced adult emergence in six habitats to (0.34 +/- 0.13) compared to (0.98 +/- 0.02) in the controls (p < 0.0001), showing a high level of habitats coverage amplification of the autodissemination event. CONCLUSION The study provides proof of principle for the autodissemination of PPF to breeding habitats by malaria vectors. These findings highlight the potential for this technique for outdoor control of malaria vectors and call for the testing of this technique in field trials

Predicting Scenarios for Successful Autodissemination of Pyriproxyfen by Malaria Vectors from Their Resting Sites to Aquatic Habitats; Description and Simulation Analysis of a Field-Parameterizable Model

Background Large-cage experiments indicate pyriproxifen (PPF) can be transferred from resting sites to aquatic habitats by Anopheles arabiensis - malaria vector mosquitoes to inhibit emergence of their own offspring. PPF coverage is amplified twice: (1) partial coverage of resting sites with PPF contamination results in far higher contamination coverage of adult mosquitoes because they are mobile and use numerous resting sites per gonotrophic cycle, and (2) even greater contamination coverage of aquatic habitats results from accumulation of PPF from multiple oviposition events. Methods and Findings Deterministic mathematical models are described that use only field-measurable input parameters and capture the biological processes that mediate PPF autodissemination. Recent successes in large cages can be rationalized, and the plausibility of success under full field conditions can be evaluated a priori. The model also defines measurable properties of PPF delivery prototypes that may be optimized under controlled experimental conditions to maximize chances of success in full field trials. The most obvious flaw in this model is the endogenous relationship that inevitably occurs between the larval habitat coverage and the measured rate of oviposition into those habitats if the target mosquito species is used to mediate PPF transfer. However, this inconsistency also illustrates the potential advantages of using a different, non-target mosquito species for contamination at selected resting sites that shares the same aquatic habitats as the primary target. For autodissemination interventions to eliminate malaria transmission or vector populations during the dry season window of opportunity will require comprehensive contamination of the most challenging subset of aquatic habitats that persist or retain PPF activity (Ux) for only one week , where Ux = 7 days). To achieve >99% contamination coverage of these habitats will necessitate values for the product of the proportional coverage of the ovipositing mosquito population with PPF contamination (CM) by the ovitrap-detectable rates of oviposition by wild mosquitoes into this subset of habitats , divided by the titre of contaminated mosquitoes required to render them unproductive , that approximately approach unity . Conclusions The simple multiplicative relationship between CM and , and the simple exponential decay effect they have upon uncontaminated aquatic habitats, allows application of this model by theoreticians and field biologists alike