Anopheles Arabiensis Research Articles

The Laboratory and Semi-Field Larvicidal Effects of Essential Oil Extracted from Feronia limonia against Anopheles arabiensis Patton.

This study intended to evaluate the larvicidal activity of Feronia limonia leaf essential oil against the wild population of Anopheles arabiensis Patton larvae in laboratory and semi-field environments. Larvae mortality was observed after 12, 24, 48, and 72 hours of exposure. In laboratory condition, the essential oil showed good larvicidal activity against An. arabiensis (LC50 = 85.61 and LC95 = 138.03 ppm (after 12 hours); LC50 = 65.53 and LC95 = 117.95 ppm (after 24 hours); LC50 = 32.18 and LC95 = 84.59 ppm (after 48 hours); LC50 = 8.03 and LC95 = 60.45 ppm (after 72 hours), while in semi-field experiments, larvicidal activity was (LC50 = 91.89 and LC95 = 134.93 ppm (after 12 hours); LC50 = 83.34 and LC95 = 109.81 ppm (after 24 hours); LC50 = 66.78 and LC95 = 109.81 (after 28 hours); LC50 = 47.64 and 90.67 ppm (after 72 hours). These results give an insight on the future use of F. limonia essential oils for mosquitoes control.

Species Composition, Seasonal Abundance, and Biting Behavior of Malaria Vectors in Rural Conhane Village, Southern Mozambique.

Malaria vector surveillance provides important data to inform the effective planning of vector control interventions at a local level. The aim of this study was to determine the species diversity and abundance, biting activity, and Plasmodium infectivity of Anopheles mosquitoes from a rural village in southern Mozambique. Human landing catches were performed monthly between December 2020 and August 2021. All collected Anopheles were identified to the species level and tested for the presence of malaria parasites. Eight Anopheles species were identified among the 1802 collected anophelines. Anopheles gambiae sensu lato (s.l.) were the most abundant (51.9%) and were represented by Anopheles quadriannulatus and Anopheles arabiensis. Anopheles funestus s.l. represented 4.5%. The biting activity of An. arabiensis was more pronounced early in the evening and outdoors, whereas that of An. funestus sensu stricto (s.s.) was more intense late in the night, with no significant differences in location. One An. funestus s.s. and one An. arabiensis, both collected outdoors, were infected with Plasmodium falciparum. The overall entomologic inoculation rate was estimated at 0.015 infective bites per person per night. The significant outdoor and early evening biting activity of An. arabiensis and An. funestus found in this village may negatively impact the effectiveness of current vector control interventions. Additional vector control tools that can target these mosquitoes are needed.

Laboratory-based efficacy evaluation of Bacillus thuringiensis var. israelensis and temephos larvicides against larvae of Anopheles stephensi in ethiopia

BackgroundMalaria, transmitted by the bite of infective female Anopheles mosquitoes, remains a global public health problem. The presence of an invasive Anopheles stephensi, capable of transmitting Plasmodium vivax and Plasmodium falciparum parasites was first reported in Ethiopia in 2016. The ecology of An. stephensi is different from that of Anopheles arabiensis, the primary Ethiopian malaria vector, and this suggests that alternative control strategies may be necessary. Larviciding may be an effective alternative strategy, but there is limited information on the susceptibility of Ethiopian An. stephensi to common larvicides. This study aimed to evaluate the efficacy of temephos and Bacillus thuringiensis var. israelensis (Bti) larvicides against larvae of invasive An. stephensi.MethodsThe diagnostic doses of two larvicides, temephos (0.25 ml/l) and Bti (0.05 mg/l) were tested in the laboratory against the immature stages (late third to early fourth stages larvae) of An. stephensi collected from the field and reared in a bio-secure insectary. Larvae were collected from two sites (Haro Adi and Awash Subuh Kilo). For each site, three hundred larvae were tested against each insecticide (as well as an untreated control), in batches of 25. The data from all replicates were pooled and descriptive statistics prepared.ResultsThe mortality of larvae exposed to temephos was 100% for both sites. Mortality to Bti was 99.7% at Awash and 100% at Haro Adi site.ConclusionsLarvae of An. stephensi are susceptible to temephos and Bti larvicides suggesting that larviciding with these insecticides through vector control programmes may be effective against An. stephensi in these localities.

Small-scale field evaluation of transfluthrin-treated eave ribbons and sandals for the control of malaria vectors in rural Tanzania

BackgroundEarly-evening and outdoor-biting mosquitoes may compromise the effectiveness of frontline malaria interventions, notably insecticide-treated nets (ITNs). This study aimed to evaluate the efficacy of low-cost insecticide-treated eave ribbons and sandals as supplementary interventions against indoor-biting and outdoor-biting mosquitoes in south-eastern Tanzania, where ITNs are already widely used.MethodsThis study was conducted in three villages, with 72 households participating (24 households per village). The households were divided into four study arms and assigned: transfluthrin-treated sandals (TS), transfluthrin-treated eave ribbons (TER), a combination of TER and TS, or experimental controls. Each arm had 18 households, and all households received new ITNs. Mosquitoes were collected using double net traps (to assess outdoor biting), CDC light traps (to assess indoor biting), and Prokopack aspirators (to assess indoor resting). Protection provided by the interventions was evaluated by comparing mosquito densities between the treatment and control arms. Additional tests were done in experimental huts to assess the mortality of wild mosquitoes exposed to the treatments or controls.ResultsTERs reduced indoor-biting, indoor-resting and outdoor-biting Anopheles arabiensis by 60%, 73% and 41%, respectively, while TS reduced the densities by 18%, 40% and 42%, respectively. When used together, TER & TS reduced indoor-biting, indoor-resting and outdoor-biting An. arabiensis by 53%, 67% and 57%, respectively. Protection against Anopheles funestus ranged from 42 to 69% with TER and from 57 to 74% with TER & TS combined. Mortality of field-collected mosquitoes exposed to TER, TS or both interventions was 56–78% for An. arabiensis and 47–74% for An. funestus.ConclusionTransfluthrin-treated eave ribbons and sandals or their combination can offer significant household-level protection against malaria vectors. Their efficacy is magnified by the transfluthrin-induced mortality, which was observed despite the prevailing pyrethroid resistance in the study area. These results suggest that TER and TS could be useful supplementary tools against residual malaria transmission in areas where ITN coverage is high but additional protection is needed against early-evening and outdoor-biting mosquitoes. Further research is needed to validate the performance of these tools in different settings, and assess their long-term effectiveness and feasibility for malaria control.

Entomological drivers of uneven malaria transmission in urban lowland areas in Bouaké, Côte d’Ivoire

BackgroundThe use of urban lowlands for agriculture contributes to the food security of city- dwellers, but promotes malaria transmission. The objective of the study was to characterize the entomological drivers of malaria transmission in two lowlands (N’Gattakro and Odiennekourani) in the city of Bouaké, Côte d’Ivoire.MethodsThe human landing catch technique was used to capture mosquitoes in houses located at the edge of two lowlands in Bouaké from February to December 2019. Cultivated surfaces were calculated monthly in both lowlands for each crop type (rice and market gardening) using images acquired by a drone. The different mosquito species were identified morphologically and by PCR analysis for the Anopheles gambiae complex. Anopheles infection by Plasmodium parasites was assessed by quantitative PCR. Mosquito diversity, biting behaviour and rhythmicity, and malaria transmission were determined in each lowland and compared.ResultsAnopheles gambiae sensu lato (s.l.) was predominant in N’Gattakro and Culex quinquefasciatus in Odiennekourani. Four Anopheles species were identified: An. gambiae s.l. and Anopheles funestus s.l. in both lowlands, Anopheles pharoensis in N’Gattakro, and Anopheles ziemanni in Odiennekourani. Within the An. gambiae complex, three species were caught: An. gambiae sensu stricto (s.s.), Anopheles coluzzii, and Anopheles arabiensis for the first time in Côte d’Ivoire (30.1%, 69.9% and 0% in N’Gattakro, and 45.1%, 52.6% and 2.4% in Odiennekourani, respectively). Anopheles gambiae s.l. species exhibited a significant exophagic behaviour in N’Gattakro (77.1% of outdoor bites versus 52.2% in Odiennekourani). In N’Gattakro, 12.6% of captures occurred before bedtime (09.00 pm) and after waking up (05.00 am), 15.1% in Odiennekourani. The mean human biting rate was higher in N’Gattakro than in Odiennekourani (61.6 versus 15.5 bites per person per night). Overall, Anopheles infection rate was 0.68%, with 0.539 and 0.029 infected bites per person per night in N’Gattakro and Odiennekourani, respectively.ConclusionThe risk of malaria in urban agricultural lowland areas is uneven. The role of agricultural developments and irrigation patterns in the production of larval habitat should be explored. The exophagic behaviour of Anopheles vectors raises the question of the residual transmission that needs to be assessed to implement appropriate control strategies.

A non-inferiority and GLP-compliant study of broflanilide IRS (VECTRON™ T500), a novel meta-diamide insecticide against Anopheles arabiensis

Management of insecticide resistance in vector control requires development and evaluation of active ingredients (AIs) with new modes of action. VECTRON™ T500 is a wettable powder formulation used for Indoor Residual Spraying (IRS) containing 50% of broflanilide as an AI. This study evaluated the efficacy of VECTRON™ T500 sprayed on blocks of different substrates (concrete, mud and plywood) against pyrethroid susceptible and resistant Anopheles gambiae sensu stricto (s.s.) strains, and wild An. arabiensis. It also assessed the efficacy of VECTRON™ T500 in experimental huts plastered with mud and concrete against wild free-flying An. arabiensis; and non-inferiority to a World Health Organization listed indoor residual spraying product Actellic® 300CS in terms of mortality in Moshi, Tanzania.Monthly cone bioassays on blocks and in experimental huts (against pyrethroid susceptible and resistant An. gambiae s.s.) were conducted over a 12-month period after spraying of VECTRON™ T500 and Actellic® CS300. Collections of wild free-flying An. arabiensis were also done in the sprayed huts. The main outcome for cone bioassays was mortality while for the wild hut trial collections, it was mortality and blood feeding inhibition. Grouped logistic regressions with random effects were used to analyse all dichotomous outcome variables from wild collections.The results showed residual efficacy of VECTRON™ T500 of at least 80% mortality was longest on concrete, followed by plywood and then mud substrates for all mosquito strains. Furthermore, VECTRON™ T500 significantly increased the likelihood of mortality (OR:> 1.37, P<0.001) in wild collections of An. arabiensis compared to Actellic® 300CS. Blood feeding was not significantly different in the wild collection of An. arabiensis between VECTRON™ T500 and Actellic® 300CS arms.These results show that VECTRON™ T500 is efficacious against pyrethroid-resistant An. gambiae s.s. and non-inferior to Actellic® 300CS. Therefore, it should be an important addition to the current arsenal of insecticides used for insecticide resistance management and vector control.

Naturally acquired antibodies to gametocyte antigens are associated with reduced transmission of Plasmodium vivax gametocytes to Anopheles arabiensis mosquitoes.

Naturally acquired antibodies may reduce the transmission of Plasmodium gametocytes to mosquitoes. Here, we investigated associations between antibody prevalence and P. vivax infectivity to mosquitoes. A total of 368 microscopy confirmed P. vivax symptomatic patients were passively recruited from health centers in Ethiopia and supplemented with 56 observations from asymptomatic P. vivax parasite carriers. Direct membrane feeding assays (DMFA) were performed to assess mosquito infectivity; for selected feeds these experiments were also performed after replacing autologous plasma with malaria naïve control serum (n=61). The prevalence of antibodies against 6 sexual stage antigens (Pvs47, Pvs48/45, Pvs230, PvsHAP2, Pvs25 and PvCelTOS) and an array of asexual antigens was determined by ELISA and multiplexed bead-based assays. Gametocyte (ρ< 0.42; p = 0.0001) and parasite (ρ = 0.21; p = 0.0001) densities were positively associated with mosquito infection rates. Antibodies against Pvs47, Pvs230 and Pvs25 were associated with 23 and 34% reductions in mosquito infection rates (p<0.0001), respectively. Individuals who showed evidence of transmission blockade in serum-replacement DMFAs (n=8) were significantly more likely to have PvsHAP2 or Pvs47 antibodies. Further studies may demonstrate causality for the observed associations, improve our understanding of the natural transmission of P. vivax and support vaccine development.

Using transfer learning and dimensionality reduction techniques to improve generalisability of machine-learning predictions of mosquito ages from mid-infrared spectra

BackgroundOld mosquitoes are more likely to transmit malaria than young ones. Therefore, accurate prediction of mosquito population age can drastically improve the evaluation of mosquito-targeted interventions. However, standard methods for age-grading mosquitoes are laborious and costly. We have shown that Mid-infrared spectroscopy (MIRS) can be used to detect age-specific patterns in mosquito cuticles and thus can be used to train age-grading machine learning models. However, these models tend to transfer poorly across populations. Here, we investigate whether applying dimensionality reduction and transfer learning to MIRS data can improve the transferability of MIRS-based predictions for mosquito ages.MethodsWe reared adults of the malaria vector Anopheles arabiensis in two insectaries. The heads and thoraces of female mosquitoes were scanned using an attenuated total reflection-Fourier transform infrared spectrometer, which were grouped into two different age classes. The dimensionality of the spectra data was reduced using unsupervised principal component analysis or t-distributed stochastic neighbour embedding, and then used to train deep learning and standard machine learning classifiers. Transfer learning was also evaluated to improve transferability of the models when predicting mosquito age classes from new populations.ResultsModel accuracies for predicting the age of mosquitoes from the same population as the training samples reached 99% for deep learning and 92% for standard machine learning. However, these models did not generalise to a different population, achieving only 46% and 48% accuracy for deep learning and standard machine learning, respectively. Dimensionality reduction did not improve model generalizability but reduced computational time. Transfer learning by updating pre-trained models with 2% of mosquitoes from the alternate population improved performance to ~ 98% accuracy for predicting mosquito age classes in the alternative population.ConclusionCombining dimensionality reduction and transfer learning can reduce computational costs and improve the transferability of both deep learning and standard machine learning models for predicting the age of mosquitoes. Future studies should investigate the optimal quantities and diversity of training data necessary for transfer learning and the implications for broader generalisability to unseen datasets.

Effect of Ivermectin® on survivorship and fertility of Anopheles arabiensis in Ethiopia: an in vitro study

BackgroundInnovative vector control tools are needed to counteract insecticide resistance and residual malaria transmission. One of such innovative methods is an ivermectin (IVM) treatment to reduce vector survival. In this study, a laboratory experiment was conducted to investigate the effect of ivermectin on survivorship, fertility and egg hatchability rate of Anopheles arabiensis in Ethiopia.MethodsAn in vitro experiment was conducted using 3–5 days old An. arabiensis adults from a colony maintained at insectary of Tropical and Infectious Diseases Research Center, Jimma University (laboratory population) and Anopheles mosquitoes reared from larvae collected from natural mosquito breeding sites (wild population). The mosquitoes were allowed to feed on cattle blood treated with different doses of ivermectin (0 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml and 80 ng/ml). During each feeding experiment, the mosquitoes were held in cages and blood-fed using a Hemotek feeder. Mortality and egg production were then recorded daily for up to 9 days. Time to death was analysed by a Cox frailty model with replicate as frailty term and source of mosquito (wild versus laboratory), treatment type (ivermectin vs control) and their interaction as categorical fixed effects. Kaplan Meier curves were plotted separately for wild and laboratory populations for a visual interpretation of mosquito survival as a function of treatment.ResultsBoth mosquito source and treatment had a significant effect on survival (P < 0.001), but their interaction was not significant (P = 0.197). Compared to the controls, the death hazard of An. arabiensis that fed on ivermectin-treated blood was 2.3, 3.5, 6.5, 11.5 and 17.9 times that of the control for the 5 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml, and 80 ng/ml dose, respectively. With respect to the number of hatched larvae, hatched pupae and emerged adults per fed mosquitoes, a significant difference was found between the control and the 5 ng/ml dose group (P < 0.001). The number of hatched larvae and pupae, and emerged adults decreased further for the 10 ng/ml dose group and falls to zero for the higher doses.ConclusionTreating cattle blood with ivermectin reduced mosquito survival, fertility, egg hatchability, larval development and adult emergence of An. arabiensis in all tested concentrations of ivermectin in both the wild and laboratory populations. Thus, ivermectin application in cattle could be used as a supplementary vector control method to tackle residual malaria transmission and ultimately achieve malaria elimination in Ethiopia.

Distance from human dwellings differentially affects the efficacy of a synthetic cattle urine odour lure to trap malaria vectors

BackgroundCost–effective outdoor–based devices for surveillance and control of outdoor mosquito vector populations can substantially improve their efficacy when baited with synthetic human and animal odours. This study aimed at assessing the dose–dependent efficacy of a previously developed synthetic cattle urine odour to lure malaria vectors, and other mosquito species, to traps placed at different distances from human dwellings outdoors.MethodsThe efficacy of the cattle urine odour lure was assessed through a 5 × 5 Latin square design, using two sets of 5 Suna traps placed at either 1.5 m or 5 m from an adjacent human dwelling, in the rural village of Sagamaganga, Tanzania. Each trap was deployed with one of four doses of the synthetic cattle urine odour blend or a solvent control (heptane). Traps were rotated daily so that each dose and control visited each position twice over a period of 20 experimental nights. The relative attractiveness of each treatment dose and control was compared using a generalized linear mixed model for each species caught.ResultsA total of 1568 mosquitoes were caught, of which 783 were anophelines and 785 were culicines. Of the anophelines, 41.6 and 58.3% were primary and secondary vector species, respectively. Unfed and fed females of the primary vector, Anopheles arabiensis, were caught dose–dependently, close to human dwellings (1.5 m), whereas unfed, fed and gravid secondary vector Anopheles pharoensis females were caught dose–dependently, but at a farther distance from the dwellings (5 m). Females of Culex spp. were caught dose–dependently in similar numbers irrespective of the distance from human dwellings.ConclusionsThis study further clarifies the factors to be considered for the implementation of outdoor trapping using the synthetic cattle urine lure to target exophilic and exophagic malaria vectors, for which efficient surveillance and control tools are currently lacking. The findings resulting from this study make significant progress in providing the needed information to overcome the regulatory obstacles to make this tool available for integrated vector management programs, including registration, as well as evaluation and regulation by the World Health Organization.

Infectivity of symptomatic Plasmodium vivax cases to different generations of wild-caught and laboratory-adapted Anopheles arabiensis using a membrane feeding assay, Ethiopia

When measuring human to mosquito transmission of Plasmodium spp., laboratory-adapted (colony) mosquitoes can be utilized. To connect transmission studies to the local epidemiology, it can be important to comprehend the relationship between infectivity in laboratory-adapted (colony) and wild-caught (wild) mosquitoes of the same species. Microscopically confirmed Plasmodium vivax cases were recruited from health facilities in Arba Minch town, and a nested polymerase chain reaction (nPCR) was used for subsequent confirmation. We performed paired membrane-feeding assays using colony An. arabiensis and three generations of wild origin An. arabiensis. Anopheles arabiensis aged 3–6 days were fed after being starved for 8–14 h. Microscopically, the oocyst development was evaluated at day 7 after feeding. Circumsporozoite proteins (CSPs) assay was carried out by enzyme-linked immunosorbent assay (ELISA). In 19 paired feeding experiments, the feeding efficiency was more than doubled in colony (median: 62.5%; interquartile range, IQR: 35–78%) than in wild mosquitoes (median: 28.5%; IQR: 17.5–40%; P < 0.001). Among the 19 P. vivax gametocyte-positive blood samples, 63.2% (n = 12) were infective to wild An. arabiensis and 73.7% (n = 14) were infective to colony An. arabiensis. The median infection rate was twice as high (26%) in the colony than in the wild (13%) An. arabiensis, although the difference was marginally insignificant (P = 0.06). Although the observed difference was not statistically significant (P = 0.19), the median number of oocysts per midgut was more than twice as high (17.8/midgut) in colony than in wild (7.2/midgut) An. arabiensis. The median feeding efficiency was 26.5% (IQR: 18–37%) in F1, 29.3% (IQR: 28–40%) in F2 and 31.2% (IQR: 30–37%) in F3 generations of wild An. arabiensis. Also, no significant difference was observed in oocyst infection rate and load between generations of wild An. arabiensis. CSP rate of P. vivax was 3.1% (3/97; 95% CI: 0.6–8.8%) in wild and 3.6% (3/84; 95% CI: 0.7–10.1%) in colony An. arabiensis. The results of the present study revealed that oocyst infection and load/midgut, and CSP rate were roughly comparable, indicating that colony mosquitoes can be employed for infectivity studies, while larger sample sizes may be necessary in future studies.

Synthesis &amp; Pharmacological Activity of Cytotoxicity Assay of Coumarins SVCM1-SVCM7

The therapeutic properties of natural and synthetic compounds against human illnesses have attracted a lot of interest. The pharmacological and biological effects of coumarins are widely exploited in medicine; they include anti-inflammatory, anticoagulant, antihypertensive, anticonvulsant, antioxidant, antibacterial, and neuroprotective properties. Signaling pathways affecting many cellular functions may also be modulated by coumarin derivates. Whether or not a coumarin has pharmacological, biochemical, or therapeutic characteristics is dependent on the nature of the replacement surrounding the coumarin core structure. The capacity to kill, repel, or otherwise impact the development of Anopheles arabiensis mosquitoes was evaluated using seven synthetic halogenated coumarins (SVCM1-SVCM7). The compounds SVCM1-SVCM7 were evaluated for their bactericidal and fungicidal activities using the disc diffusion method.

Seasonal variation in abundance and blood meal sources of primary and secondary malaria vectors within Kilombero Valley, Southern Tanzania

BackgroundExisting control tools have significantly reduced malaria over the past two decades. However, progress has been stalled due to increased resistance in primary vectors and the increasing role of secondary vectors. This study aimed to investigate the impact of seasonal change on primary and secondary vector abundance and host preference. Understanding the impact of seasonal dynamics of primary and secondary vectors on disease transmission will inform effective strategies for vector management and control.MethodsVector abundance was measured through longitudinal collection of mosquitoes, conducted monthly during the wet and dry seasons, in Sagamaganga, a village in the Kilombero Valley, Tanzania. Mosquitoes were collected indoors using CDC light traps and backpack aspirators, and outdoors using resting buckets baited with cattle urine. In addition, a direct measure of host preference was taken monthly using human- and cattle-baited mosquito electrocuting traps. A host census was conducted to provide an indirect measure of host preference together with monthly blood meal source analysis. All collected mosquitoes were assayed for Plasmodium sporozoites.ResultsA total of 2828 anophelines were collected, of which 78.5% and 21.4%, were primary and secondary vectors, respectively. The abundance of the primary vectors, Anopheles arabiensis and Anopheles funestus, and of the secondary vectors varied seasonally. Indirect measures of host preference indicated that all vectors varied blood meal choice seasonally, with the direct measure confirming this for An. arabiensis. All anopheline mosquitoes tested negative for sporozoites.ConclusionsAt the study location, the abundance of both primary and secondary vectors changed seasonally. Indirect and direct measures of host preference demonstrated that An. arabiensis varied from being zoophilic to being more opportunistic during the wet and dry seasons. A similar trend was observed for the other vectors.Graphical

Impact of different mosquito collection methods on indicators of Anopheles malaria vectors in Uganda

BackgroundMethods used to sample mosquitoes are important to consider when estimating entomologic metrics. Human landing catches (HLCs) are considered the gold standard for collecting malaria vectors. However, HLCs are labour intensive, can expose collectors to transmission risk, and are difficult to implement at scale. This study compared alternative methods to HLCs for collecting Anopheles mosquitoes in eastern Uganda.MethodsBetween June and November 2021, mosquitoes were collected from randomly selected households in three parishes in Tororo and Busia districts. Mosquitoes were collected indoors and outdoors using HLCs in 16 households every 4 weeks. Additional collections were done indoors with prokopack aspirators, and outdoors with pit traps, in these 16 households every 2 weeks. CDC light trap collections were done indoors in 80 households every 4 weeks. Female Anopheles mosquitoes were identified morphologically and Anopheles gambiae sensu lato were speciated using PCR. Plasmodium falciparum sporozoite testing was done with ELISA.ResultsOverall, 4,891 female Anopheles were collected, including 3,318 indoors and 1,573 outdoors. Compared to indoor HLCs, vector density (mosquitoes per unit collection) was lower using CDC light traps (4.24 vs 2.96, density ratio [DR] 0.70, 95% CIs 0.63–0.77, p < 0.001) and prokopacks (4.24 vs 1.82, DR 0.43, 95% CIs 0.37–0.49, p < 0.001). Sporozoite rates were similar between indoor methods, although precision was limited. Compared to outdoor HLCs, vector density was higher using pit trap collections (3.53 vs 6.43, DR 1.82, 95% CIs 1.61–2.05, p < 0.001), while the sporozoite rate was lower (0.018 vs 0.004, rate ratio [RR] 0.23, 95% CIs 0.07–0.75, p = 0.008). Prokopacks collected a higher proportion of Anopheles funestus (75.0%) than indoor HLCs (25.8%), while pit traps collected a higher proportion of Anopheles arabiensis (84.3%) than outdoor HLCs (36.9%).ConclusionIn this setting, the density and species of mosquitoes collected with alternative methods varied, reflecting the feeding and resting characteristics of the common vectors and the different collection approaches. These differences could impact on the accuracy of entomological indicators and estimates of malaria transmission, when using the alternative methods for sampling mosquitos, as compared to HLCs.

Manure and mosquitoes: life history traits of two malaria vector species enhanced by larval exposure to cow dung, whilst chicken dung has a strong negative effect

BackgroundMalaria vectors have a strong ecological association with rice agroecosystems, which can provide abundant aquatic habitats for larval development. Climate-adapted rice cultivation practices, such as the System of Rice Intensification (SRI), are gaining popularity in malaria-endemic countries seeking to expand rice production; however, the potential impact of these practices on vector populations has not been well characterised. In particular, SRI encourages the use of organic fertilisers (OFs), such as animal manures, as low-cost and environmentally friendly alternatives to industrially produced inorganic fertilisers. We therefore set out to understand the effects of two common manure-based OFs on the life history traits of two major African malaria vectors, Anopheles arabiensis and Anopheles gambiae sensu stricto (s.s.).MethodsLarvae of An. arabiensis and An. gambiae s.s. were reared from first instar to emergence in water containing either cow or chicken dung at one of four concentrations (0.25, 0.5, 0.75, and 1.0 g/100 ml), or in a clean water control. Their life history traits were recorded, including survival, development rate, adult production, and adult wing length.ResultsExposure to cow dung significantly increased the development rate of An. gambiae s.s. independent of concentration, but did not affect the overall survival and adult production of either species. Chicken dung, however, significantly reduced survival and adult production in both species, with a greater effect as concentration increased. Interestingly, An. arabiensis exhibited a relative tolerance to the lowest chicken dung concentration, in that survival was unaffected and adult production was not reduced to the same extent as in An. gambiae s.s. The effects of chicken dung on development rate were less clear in both species owing to high larval mortality overall, though there was some indication that it may reduce development rate. Adult wing lengths in males and females increased with higher concentrations of both cow and chicken dung.ConclusionsOur findings suggest that manure-based OFs significantly alter the life history traits of An. gambiae s.s. and An. arabiensis. In both species, exposure to cow dung may improve fitness, whereas exposure to chicken dung may reduce it. These findings have implications for understanding vector population dynamics in rice agroecosystems and may inform the use of OFs in SRI, and rice agriculture more widely, to avoid their adverse effects in enhancing vector fitness.Graphical

Countrywide insecticide resistance monitoring and first report of the presence of the L1014S knock down resistance in Niger, West Africa

BackgroundMass distribution of insecticide-treated nets (ITNs) is the principal malaria vector control strategy adopted by Niger. To better inform on the most appropriate ITN to distribute, the National Malaria Control Programme (NMCP) of Niger and its partners, conducted insecticide resistance monitoring in selected sites across the country.MethodsThe susceptibility of Anopheles gambiae sensu lato (s.l.) to chlorfenapyr and pyrethroid insecticides was investigated in a total of sixteen sites in 2019 and 2020, using 2–5-day-old adults reared from wild collected larvae per site. The susceptibility status, pyrethroid resistance intensity at 5 and 10 times the diagnostic concentrations, and piperonyl butoxide (PBO) synergism with diagnostic concentrations of deltamethrin, permethrin and alpha-cypermethrin were assessed using WHO bioassays. Two doses (100 and 200 µg/bottle) of chlorfenapyr were tested using the CDC bottle assay method. Species composition and allele frequencies for knock-down resistance (kdr-L1014F and L1014S) and acetylcholinesterase (ace-1 G119S) mutations were further characterized using polymerase chain reaction (PCR).ResultsHigh resistance intensity to all pyrethroids tested was observed in all sites except for alpha-cypermethrin in Gaya and Tessaoua and permethrin in Gaya in 2019 recording moderate resistance intensity. Similarly, Balleyara, Keita and Tillabery yielded moderate resistance intensity for alpha-cypermethrin and deltamethrin, and Niamey V low resistance intensity against deltamethrin and permethrin in 2020. Pre-exposure to PBO substantially increased susceptibility with average increases in mortality between 0 and 70% for tested pyrethroids. Susceptibility to chlorfenapyr (100 µg/bottle) was recorded in all sites except in Tessaoua and Magaria where susceptibility was recorded at the dose of 200 µg/bottle. Anopheles coluzzii was the predominant malaria vector species in most of the sites followed by An. gambiae sensu stricto (s.s.) and Anopheles arabiensis. The kdr-L1014S allele, investigated for the first time, was detected in the country. Both kdr-L1014F (frequencies [0.46–0.81]) and L1014S (frequencies [0.41–0.87]) were present in all sites while the ace-1 G119S was between 0.08 and 0.20.ConclusionThe data collected will guide the NMCP in making evidence-based decisions to better adapt vector control strategies and insecticide resistance management in Niger, starting with mass distribution of new generation ITNs such as interceptor G2 and PBO ITNs.

Characterizing pyrethroid resistance and mechanisms in Anopheles gambiae (s.s.) and Anopheles arabiensis from 11 districts in Uganda

Insecticide resistance threatens recent progress on malaria control in Africa. To characterize pyrethroid resistance in Uganda, Anopheles gambiae (s.s.) and Anopheles arabiensis were analyzed from 11 sites with varied vector control strategies. Mosquito larvae were collected between May 2018 and December 2020. Sites were categorized as receiving no indoor-residual spraying (‘no IRS’, n ​= ​3); where IRS was delivered from 2009 to 2014 and in 2017 and then discontinued (‘IRS stopped’, n ​= ​4); and where IRS had been sustained since 2014 (‘IRS active’, n ​= ​4). IRS included bendiocarb, pirimiphos methyl and clothianidin. All sites received long-lasting insecticidal nets (LLINs) in 2017. Adult mosquitoes were exposed to pyrethroids; with or without piperonyl butoxide (PBO). Anopheles gambiae (s.s.) and An. arabiensis were identified using PCR. Anopheles gambiae (s.s.) were genotyped for Vgsc-995S/F, Cyp6aa1, Cyp6p4-I236M, ZZB-TE, Cyp4j5-L43F and Coeae1d, while An. arabiensis were examined for Vgsc-1014S/F. Overall, 2753 An. gambiae (s.l.), including 1105 An. gambiae (s.s.) and 1648 An. arabiensis were evaluated. Species composition varied by site; only nine An. gambiae (s.s.) were collected from ‘IRS active’ sites, precluding species-specific comparisons. Overall, mortality following exposure to permethrin and deltamethrin was 18.8% (148/788) in An. gambiae (s.s.) and 74.6% (912/1222) in An. arabiensis. Mortality was significantly lower in An. gambiae (s.s.) than in An. arabiensis in ‘no IRS’ sites (permethrin: 16.1 vs 67.7%, P ​< ​0.001; deltamethrin: 24.6 vs 83.7%, P ​< ​0.001) and in ‘IRS stopped’ sites (permethrin: 11.3 vs 63.6%, P ​< ​0.001; deltamethrin: 25.6 vs 88.9%, P ​< ​0.001). When PBO was added, mortality increased for An. gambiae (s.s.) and An. arabiensis. Most An. gambiae (s.s.) had the Vgsc-995S/F mutation (95% frequency) and the Cyp6p4-I236M resistance allele (87%), while the frequency of Cyp4j5 and Coeae1d were lower (52% and 55%, respectively). Resistance to pyrethroids was widespread and higher in An. gambiae (s.s.). Where IRS was active, An. arabiensis dominated. Addition of PBO to pyrethroids increased mortality, supporting deployment of PBO LLINs. Further surveillance of insecticide resistance and assessment of associations between genotypic markers and phenotypic outcomes are needed to better understand mechanisms of pyrethroid resistance and to guide vector control.

Residual efficacy of SumiShield™ 50WG for indoor residual spraying in Ethiopia

BackgroundThe rate of decay of the biological efficacy of insecticides used for indoor residual spraying (IRS) is an important factor when making decisions on insecticide choice for national malaria control programmes. A key roadblock to IRS programme is insecticide resistance. If resistance is detected to most of the existing insecticides used for IRS (DDT, pyrethroids, organophosphates and carbamates), the logical next choice could be neonicotinoid insecticides, as pyrethroids are used to treat nets. SumiShield™ 50WG belongs to the neonicotinoid class of insecticides and has shown promising results in several phase I, II and III trials in different settings. The aim of this study was to assess the persistence of SumiShield™ 50WG by spraying on different wall surfaces and determine its decay rates over time in Ethiopia.MethodsFive huts with different wall surface types (mud, dung, paint and cement) which represented the Ethiopian house wall surfaces were used to evaluate the residual efficacy of SumiShield™ 50WG. Actellic 300CS sprayed on similar wall surfaces of another five huts was used as a comparator insecticide and two huts sprayed with water were used as a control. All huts were sprayed uniformly by an experienced spray operator; non-stop starting from the door and moving clockwise to cover the entire wall surface of the hut. The treatments were assigned to huts randomly. The residual efficacy of the insecticide formulations was evaluated against a susceptible insectary-reared population of Anopheles arabiensis using WHO cone bioassays.ResultsSumiShield™ 50WG resulted in mortality rates of over 80% at 120 h post-exposure on all surface types for up to nine months post-spray, while Actellic 300CS yielded mortality rates of over 80% for eight months after spray.ConclusionsThe results of this trial demonstrated that the residual efficacy of SumiShield™ 50WG extends up to nine months on all treated wall surface types. The long-lasting residual efficacy and unique mode of action of the SemiShield™ 50WG shows that it could be an ideal product to be considered as a potential candidate insecticide formulation for IRS in malaria endemic countries such as Ethiopia or other sub-Saharan countries where the transmission season lasts up to four months or longer.

Entomological impact of mass administration of ivermectin and dihydroartemisinin-piperaquine in The Gambia: a cluster-randomized controlled trial

BackgroundVector control interventions in sub-Saharan Africa rely on insecticide-treated nets and indoor residual spraying. Insecticide resistance, poor coverage of interventions, poor quality nets and changes in vector behavior threaten the effectiveness of these interventions and, consequently, alternative tools are needed. Mosquitoes die after feeding on humans or animals treated with ivermectin (IVM). Mass drug administration (MDA) with IVM could reduce vector survival and decrease malaria transmission. The entomological impact of MDA of combined IVM and dihydroartemisinin-piperaquine was assessed in a community-based, cluster-randomized trial.MethodsA cluster-randomized trial was implemented in 2018 and 2019 in 32 villages in the Upper River Region, The Gambia. The with the inhabitants of 16 intervention villages eligible to receive three monthly rounds of MDA at the beginning of the malaria transmission season. Entomological surveillance with light traps and human landing catches (HLC) was carried out during a 7- to 14-day period after each round of MDA, and then monthly until the end of the year. The mosquitocidal effect of IVM was determined by direct membrane feeding assays.ResultsOf the 15,017 mosquitoes collected during the study period, 99.65% (n = 14,965) were Anopheles gambiae sensu lato (An. gambiae s.l.), comprising Anopheles arabiensis (56.2%), Anopheles coluzzii (24.5%), Anopheles gambiae sensu stricto (An. gembiae s.s.; 16.0%) and Anopheles funestus sensu lato (An. funestus s.l.; 0.35%). No effect of the intervention on vector parity was observed. Vector density determined on light trap collections was significantly lower in the intervention villages in 2019 (adjusted incidence rate ratio: 0.39; 95% confidence interval [CI]: 0.20, 0.74; P = 0.005) but not in 2018. However, vector density determined in HLC collections was similar in both the intervention and control villages. The entomological inoculation rate was significantly lower in the intervention villages than in the control villages (odds ratio: 0.36, 95% CI: 0.19, 0.70; P = 0·003). Mosquito mortality was significantly higher when blood fed on IVM-treated individuals up to 21 days post-treatment, particularly in adults and individuals with a higher body mass index.ConclusionMass drug administration with IVM decreased vector density and the entomological inoculation rate while the effect on vector parity was less clear. Survival of mosquitoes fed on blood collected from IVM-treated individuals was significantly lower than that in mosquitoes which fed on controls. The influence of host characteristics on mosquito survivorship indicated that dose optimization could improve IVM efficacy. Future detailed entomological evaluation trials in which IVM is administered as stand-alone intervention may elucidate the contribution of this drug to the observed reduction in transmission.Graphical

Human and vector behaviors determine exposure to Anopheles in Namibia

BackgroundAlthough the Republic of Namibia has significantly reduced malaria transmission, regular outbreaks and persistent transmission impede progress towards elimination. Towards an understanding of the protective efficacy, as well as gaps in protection, associated with long-lasting insecticidal nets (LLINs), human and Anopheles behaviors were evaluated in parallel in three malaria endemic regions, Kavango East, Ohangwena and Zambezi, using the Entomological Surveillance Planning Tool to answer the question: where and when are humans being exposed to bites of Anopheles mosquitoes?MethodsSurveillance activities were conducted during the malaria transmission season in March 2018 for eight consecutive nights. Four sentinel structures per site were selected, and human landing catches and human behavior observations were consented to for a total of 32 collection nights per site. The selected structures were representative of local constructions (with respect to building materials and size) and were at least 100 m from each other. For each house where human landing catches were undertaken, a two-person team collected mosquitoes from 1800 to 0600 hours.ResultsSurveillance revealed the presence of the primary vectors Anopheles arabiensis, Anopheles gambiae sensu stricto (s.s.) and Anopheles funestus s.s., along with secondary vectors (Anopheles coustani sensu lato and Anopheles squamosus), with both indoor and outdoor biting behaviors based on the site. Site-specific human behaviors considerably increased human exposure to vector biting. The interaction between local human behaviors (spatial and temporal presence alongside LLIN use) and vector behaviors (spatial and temporal host seeking), and also species composition, dictated where and when exposure to infectious bites occurred, and showed that exposure was primarily indoors in Kavango East (78.6%) and outdoors in Ohangwena (66.7%) and Zambezi (81.4%). Human behavior-adjusted exposure was significantly different from raw vector biting rate.ConclusionsIncreased LLIN use may significantly increase protection and reduce exposure to malaria, but may not be enough to eliminate the disease, as gaps in protection will remain both indoors (when people are awake and not using LLINs) and outdoors. Alternative interventions are required to address these exposure gaps. Focused and question-based operational entomological surveillance together with human behavioral observations may considerably improve our understanding of transmission dynamics as well as intervention efficacy and gaps in protection.Graphical