Novel sampling methods for monitoring Anopheles arabiensis from Eritrea

Background Studies comparing novel collection methods for host seeking and resting mosquitoes A. arabiensis were undertaken in a village in Eritrea. Techniques included an odor baited trap, a novel tent-trap, human landing collection and three methods of resting collection. A technique for the collection of mosquitoes exiting vegetation is also described. Pre-gravid rates were determined by dissection of host seeking insects and post-prandial egg development among insects collected resting. Results Overall 5,382 host-seeking, 2,296 resting and 357 A. arabiensis exiting vegetation were collected. The Furvela tent-trap was the most efficient, risk-free method for the collection of outdoor host-seeking insects, whilst the Suna trap was the least effective method. Mechanical aspirators (the CDC backpack or the Prokopack aspirator) were superior to manual aspiration in a dark shelter but there was no advantage over manual aspiration in a well-lit one. An estimated two-thirds of newly-emerged mosquitoes went through a pre-gravid phase, feeding twice before producing eggs. Mosquitoes completed gonotrophic development in a dark shelter but left a well-lit shelter soon after feeding. One blood-fed female marked in the village was recaptured 2 days after release exiting vegetation close to the oviposition site and another, shortly after oviposition, attempting to feed on a human host 3 days after release. Exit rates of males from vegetation peaked 3 min after the initial male had left. Unfed and gravid females exited approximately 6 min after the first males. Conclusions Furvela tent-traps are suitable for the collection of outdoor biting A. arabiensis in Eritrea whilst the Prokopack sampler is the method of choice for the collection of resting insects. Constructing well-lit, rather than dark, animal shelters, may encourage otherwise endophilic mosquitoes to leave and so reduce their survival and hence their vectorial capacity.

Comparison of four outdoor mosquito trapping methods as potential replacements for human landing catches in western Kenya

Introduction Longitudinal monitoring of outdoor-biting malaria vector populations is becoming increasingly important in understanding the dynamics of residual malaria transmission. However, the human landing catch (HLC), the gold standard for measuring human biting rates indoors and outdoors, is costly and raises ethical concerns related to increased risk of infectious bites among collectors. Consequently, routine data on outdoor-feeding mosquito populations are usually limited because of the lack of a scalable tool with similar sensitivity to outdoor HLC. Methodology The Anopheles trapping sensitivity of four baited proxy outdoor trapping methods—Furvela tent trap (FTT), host decoy trap (HDT), mosquito electrocuting traps (MET) and outdoor CDC light traps (OLT)—was assessed relative to HLC in a 5 × 5 replicated Latin square conducted over 25 nights in two villages of western Kenya. Indoor CDC light trap (ILT) was run in one house in each of the compounds with outdoor traps, while additional non-Latin square indoor and outdoor HLC collections were performed in one of the study villages. Results The MET, FTT, HDT and OLT sampled approximately 4.67, 7.58, 5.69 and 1.98 times more An. arabiensis compared to HLC, respectively, in Kakola Ombaka. Only FTT was more sensitive relative to HLC in sampling An. funestus in Kakola Ombaka (RR = 5.59, 95% CI 2.49–12.55, P < 0.001) and Masogo (RR = 4.38, 95% CI 1.62–11.80, P = 0.004) and in sampling An. arabiensis in Masogo (RR = 5.37, 95% CI 2.17–13.24, P < 0.001). OLT sampled significantly higher numbers of An. coustani in Kakola Ombaka (RR = 3.03, 95% CI 1.65–5.56, P < 0.001) and Masogo (RR = 2.88, 95% CI 1.15–7.22, P = 0.02) compared to HLC. OLT, HLC and MET sampled mostly An. coustani, FTT had similar proportions of An. funestus and An. arabiensis, while HDT sampled predominantly An. arabiensis in both villages. FTT showed close correlation with ILT in vector abundance for all three species at both collection sites. Conclusion FTT and OLT are simple, easily scalable traps and are potential replacements for HLC in outdoor sampling of Anopheles mosquitoes. However, the FTT closely mirrored indoor CDC light trap in mosquito indices and therefore may be more of an indoor mimic than a true outdoor collection tool. HDT and MET show potential for sampling outdoor host-seeking mosquitoes. However, the traps as currently designed may not be feasible for large-scale, longitudinal entomological monitoring. Therefore, the baited outdoor CDC light trap may be the most appropriate tool currently available for assessment of outdoor-biting and malaria transmission risk. Graphic abstract

Heritability and phenotypic plasticity of biting time behaviors in the major African malaria vector Anopheles arabiensis

Use of Insecticide Treated Nets for malaria control has been associated with shifts in mosquito vector feeding behavior including earlier and outdoor biting on humans. The relative contribution of phenotypic plasticity and heritability to these behavioral shifts is unknown. Elucidation of the mechanisms behind these shifts is crucial for anticipating impacts on vector control. We used a novel portable semi-field system (PSFS) to experimentally measure heritability of biting time in the malaria vector Anopheles arabiensis in Tanzania. In PSFS assays, the biting time of F2 offspring (early: 18:00-21:00, mid: 22:00-04:00 or late: 05:00-07:00) was significantly associated with that of their wild-caught F0 grandmothers, corresponding to an estimated heritability of 0.30 (95% CI: 0.20, 0.41). F2 from early-biting F0 were more likely to bite early than F2 from mid or late late-biting F0. Similarly, the probability of biting late was higher in F2 derived from mid and late-biting F0 than from early-biting F0. Our results indicate that variation in biting time is attributable to additive genetic variation. Selection can therefore act efficiently on mosquito biting times, highlighting the need for control methods that target early and outdoor biting mosquitoes.

Towards Safer and More Sustainable Systems for Monitoring Exophagic Vectors of Malaria in Western Kenya

Introduction. Longitudinal monitoring of outdoor-biting malaria vector populations is becoming increasingly important in understanding the dynamics of residual malaria transmission. However, the human landing catch (HLC), the gold standard for measuring human biting rates indoors and outdoors, is costly and raises ethical concerns related to increased risk of infective bites among collectors. Consequently, routine data on outdoor-feeding mosquito populations are usually limited due to the lack of a scalable tool with similar sensitivity to outdoor HLC. Methodology. The Anopheles trapping sensitivity of four baited proxy outdoor trapping methods—Furvela tent trap (FTT), host decoy trap (HDT), mosquito electrocuting traps (MET) and outdoor CDC light traps (OLT)—was assessed relative to HLC in a 5x5 replicated Latin square conducted over 25 nights in two villages of western Kenya. Indoor CDC light trap (ILT) was run in one house in each of the compounds with outdoor traps, while additional non-Latin square indoor and outdoor HLC collections were performed in one of the study villages. Results. The MET, FTT, HDT and OLT sampled approximately 4.67, 7.58, 5.69 and 1.98 times more An. arabiensis compared to HLC, respectively, in Kakola Ombaka. Only FTT was more sensitive relative to HLC in sampling of An. funestus in Kakola Ombaka (RR=5.59, 95%CI: 2.49-12.55, P < 0.001) and Masogo (RR=4.38, 95%CI: 1.62-11.80, P = 0.004) and in sampling An. arabiensis in Masogo (RR=5.37, 95%CI: 2.17-13.24, P < 0.001). OLT sampled significantly higher numbers of An. coustani in Kakola Ombaka (RR=3.03, 95%CI: 1.65-5.56, P < 0.001) and Masogo (RR=2.88, 95%CI: 1.15-7.22, P=0.02) compared to HLC. OLT, HLC and MET sampled mostly An. coustani, FTT had similar proportions of An. funestus and An. arabiensis, while HDT sampled predominantly An. arabiensis in both villages. FTT showed close correlation with ILT in vector abundance for all three species at both collection sites. Conclusion. FTT and OLT are simple, easily scalable traps and are potential replacements for HLC in outdoor sampling of Anopheles mosquitoes. However, the FTT closely mirrored indoor CDC light trap in mosquito indices and therefore may be more of an indoor mimic than a true outdoor collection tool. HDT and MET show potential for sampling outdoor host seeking mosquitoes. However, the traps as currently designed may not be feasible for large scale, longitudinal entomological monitoring. Therefore, the baited outdoor CDC light trap may be the most appropriate tool currently available for assessment of outdoor-biting and malaria transmission risk.

Assessing the impact of low-technology emanators alongside long-lasting insecticidal nets to control malaria

Malaria control in sub-Saharan Africa relies on the widespread use of long-lasting insecticidal nets (LLINs) or the indoor residual spraying of insecticide. Disease transmission may be maintained even when these indoor interventions are universally used as some mosquitoes will bite in the early morning and evening when people are outside. As countries seek to eliminate malaria, they can target outdoor biting using new vector control tools such as spatial repellent emanators, which emit airborne insecticide to form a protective area around the user. Field data are used to incorporate a low-technology emanator into a mathematical model of malaria transmission to predict its public health impact across a range of scenarios. Targeting outdoor biting by repeatedly distributing emanators alongside LLINs increases the chance of elimination, but the additional benefit depends on the level of anthropophagy in the local mosquito population, emanator effectiveness and the pre-intervention proportion of mosquitoes biting outdoors. High proportions of pyrethroid-resistant mosquitoes diminish LLIN impact because of reduced mosquito mortality. When mosquitoes are highly anthropophagic, this reduced mortality leads to more outdoor biting and a reduced additional benefit of emanators, even if emanators are assumed to retain their effectiveness in the presence of pyrethroid resistance. Different target product profiles are examined, which show the extra epidemiological benefits of spatial repellents that induce mosquito mortality. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.

Veterinary endectocides for malaria control and elimination: prospects and challenges

Residual transmission is the persistence of malaria transmission after scale-up of appropriate vector control tools and is one of the key challenges for malaria elimination today. Although long associated with outdoor biting, other mosquito behaviours such as partly feeding upon animals contribute greatly to sustaining transmission. Peri-domestic livestock can be used as decoy to protect humans from blood-seeking vectors but this approach often leads to an increased malaria risk in a phenomenon known as zoopotentiation. Treating the said livestock with drugs capable of killing intestinal parasites as well as mosquitoes that feed upon them has the potential to tackle malaria through a previously unexplored mechanism. The advantages and challenges associated with this approach are briefly discussed here. Numerous references are purposely provided. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.

Single blinded semi-field evaluation of MAÏA® topical repellent ointment compared to unformulated 20% DEET against Anopheles gambiae, Anopheles arabiensis and Aedes aegypti in Tanzania

Background N, N-Diethyl-3-methylbenzamide (DEET) topical mosquito repellents are effective personal protection tools. However, DEET-based repellents tend to have low consumer acceptability because they are cosmetically unappealing. More attractive formulations are needed to encourage regular user compliance. This study evaluated the protective efficacy and protection duration of a new topical repellent ointment containing 15% DEET, MAÏA® compared to 20% DEET in ethanol using malaria and dengue mosquito vectors in Bagamoyo Tanzania.Methods Fully balanced 3x3 Latin square design studies were conducted in large semi-field chambers using laboratory strains of Anopheles gambiae sensu stricto, Anopheles arabiensis and Aedes aegypti. Human volunteers applied either MAÏA® ointment, 20% DEET or ethanol to their lower limbs six hours before the start of tests. Approximately 100 mosquitoes per strain per replicate were released inside each chamber, with 25 mosquitoes released at regular intervals during the collection period to maintain adequate biting pressure throughout the test. Volunteers recaptured mosquitoes landing on their lower limbs for six hours over a period of six to 12-hours post-application of repellents. Data analysis was conducted using mixed-effects logistic regression.Results The protective efficacy of MAÏA® and 20% DEET was not different for each of the mosquito strains: 95.9% vs 97.4% against An. gambiae (OR=1.53 [95% CI: 0.93–2.51] p=0.091); 96.8% vs 97.2% against An. arabiensis (OR =1.08 [95% CI: 0.66 –1.77] P=0.757); 93.1% vs 94.6% against Ae. aegypti (OR=0.76 [95% CI: 0.20-2.80] p=0.675). Average complete protection time (CPT) of MAÏA® and that of DEET was similar for each of the mosquito strains: 571.6 minutes (95% CI: 558.3-584.8) vs 575.0 minutes (95% CI: 562.1-587.9) against An. gambiae; 585.6 minutes (95% CI: 571.4-599.8) vs 580.9 minutes (95% CI: 571.1-590.7) against An. arabiensis; 444.1 minutes (95% CI: 401.8-486.5) vs 436.9 minutes (95% CI: 405.2-468.5) against Ae. aegypti.Conclusions MAÏA® repellent ointment provides complete protection for 9 hours against both An. gambiae and An. arabiensis, and 7 hours against Ae. aegypti similar to 20% DEET (in ethanol). MAÏA® repellent ointment can be recommended to be used as a tool for prevention of outdoor biting mosquitoes in tropical locations as it protects for more than 6 hours.

Field Evaluation of a Spatial Repellent Emanation Vest for Personal Protection Against Outdoor Biting Mosquitoes

Exophilic vectors are an important contributor to residual malaria transmission. Wearable spatial repellents (SR) can potentially provide personal protection in early evening hours before people retire indoors. An SR prototype for passive delivery of transfluthrin (TFT) for protecting humans against nocturnal mosquitoes in Kanchanaburi, western Thailand, is evaluated. A plastic polyethylene terephthalate (PET) sheet (676 cm2) treated with 55-mg TFT (TFT-PET), attached to the back of short-sleeve vest worn by human collector, was evaluated under semifield and outdoor conditions. Field-caught, nonblood-fed female Anopheles minimus s.l. were released in a 40 m length, semifield screened enclosure. Two collectors positioned at opposite ends conducted 12-h human-landing collections (HLC). The outdoor experiment was conducted between treatments among four collectors at four equidistant positions who performed HLC. Both trials were conducted for 30 consecutive nights. TFT-PET provided 67% greater protection (P &lt; 0.001) for 12 h compared with unprotected control, a threefold reduction in the attack. In outdoor trials, TFT-PET provided only 16% protection against An. harrisoni Harbach & Manguin (Diptera: Culicidae) compared with unprotected collector (P = 0.0213). The TFT-PET vest reduced nonanophelines landing by 1.4-fold compared with the PET control with a 29% protective efficacy. These findings suggest that TFT-PET had diminished protective efficacy in an open field environment. Nonetheless, the concept of a wearable TFT emanatory device has the potential for protecting against outdoor biting mosquitoes. Further development of portable SR tools is required, active ingredient selection and dose optimization, and more suitable device design and materials for advancing product feasibility.

Single blinded semi-field evaluation of MAÏA® topical repellent ointment compared to unformulated 20% DEET against Anopheles gambiae s.s., Anopheles arabiensis and Aedes aegypti in Tanzania

Background: N,N-Diethyl-3-methylbenzamide (DEET) topical mosquito repellents are effective personal protection tools. However, DEET-based repellents tend to have low consumer acceptability because they are cosmetically unappealing. More attractive formulations are needed to encourage regular user compliance. This study evaluated the protective efficacy and protection duration of a new topical repellent ointment containing 15% DEET, MAÏA® compared to 20% DEET in ethanol using malaria and dengue mosquito vectors in Bagamoyo Tanzania.Methods: Fully balanced 3x3 Latin square design studies were conducted in large semi-field chambers using laboratory strains of Anopheles gambiae s.s, An. arabiensis and Aedes aegypti. Human volunteers applied either MAÏA® ointment, 20% DEET or ethanol to their lower limbs six hours before the start of tests. Approximately 100 mosquitoes per strain per replicate were released inside each chamber, with 25 mosquitoes released at regular intervals during the collection period to maintain adequate biting pressure throughout the test. Volunteers recaptured mosquitoes landing on their lower limbs for six hours over a period of six to 12-hours post-application of repellents. Data analysis was conducted using mixed-effects logistic regression.Results: The protective efficacy of MAÏA® and 20% DEET was not different for each of the mosquito strains: 95.9% vs 97.4% against An. gambiae (OR=1.53 [95% CI: 0.93–2.51] p=0.091); 96.8% vs 97.2% against An. arabiensis (OR =1.08 [95% CI: 0.66 –1.77] P=0.757); 93.1% vs 94.6% against Ae. aegypti (OR=0.76 [95% CI: 0.20-2.80] p=0.675). Average complete protection time (CPT) of MAÏA® and that of DEET was similar for each of the mosquito strains: 571.6 minutes (95% CI: 558.3-584.8) vs 575.0 minutes (95% CI: 562.1-587.9) against An. gambiae; 585.6 minutes (95% CI: 571.4-599.8) vs 580.9 minutes (95% CI: 571.1-590.7) against An. arabiensis; 444.1 minutes (95% CI: 401.8-486.5) vs 436.9 minutes (95% CI: 405.2-468.5) against Ae. aegypti.Conclusions: MAÏA® repellent ointment provides complete protection for 9 hours against both An. gambiae and An. arabiensis, and 7 hours against Ae. aegypti similar to 20% DEET (in ethanol). MAÏA® repellent ointment can be recommended to be used as a tool for prevention of outdoor biting mosquitoes in tropical locations as it protects for more than 6 hours.