Rapid entomological assessment in eight high malaria endemic regencies in Papua Province revealed the presence of indoor and outdoor malaria transmissions.

Malaria in eastern Indonesia remains high despite significant reduction and elimination in other parts of the country. A rapid entomological assessment was conducted in eight high malaria endemic regencies of Papua Province, Indonesia, to expedite malaria elimination efforts in this region. This study aims to characterize specific, actionable endpoints toward understanding where and when malaria transmission is happening, where interventions may function best, and identify gaps in protection that result in continued transmission. The entomological assessment included identifying potential vectors through human landing catch (HLC), indoor morning and night resting collections, identification of larval sites through surveillance of water bodies, and vector incrimination toward understanding exposure to malaria transmission. Human landing catches (HLCs) and larval collections identified 10 Anopheles species, namely Anopheles koliensis, Anopheles punctulatus, Anopheles farauti, Anopheles hinesorum, Anopheles longirostris, Anopheles peditaeniatus, Anopheles tesselatus, Anopheles vagus, Anopheles subpictus and Anopheles kochi. The most common and abundant species found overall were An. koliensis and An. punctulatus, while An. farauti was found in large numbers in the coastal areas of Mimika and Sarmi Regencies. Vector incrimination on Anopheles collected from HLCs and night indoor resting demonstrated that An. koliensis and An. punctulatus carried Plasmodium in Keerom, Jayapura, and Sarmi Regencies. Analysis of HLCs for the most common species revealed that the An. koliensis and An. punctulatus, bite indoors and outdoors at equal rates, while An. farauti predominantly bite outdoors. Larval surveillance demonstrated that most water bodies in and surrounding residential areas contained Anopheles larvae. This study demonstrated indoor and outdoor exposure to mosquito bites and gaps in protection, enabling exposure to infectious bites in all regencies. This explains why current malaria control efforts focusing on indoor protection have failed to substantially reduce malaria incidence in the region. Optimization of insecticide-treated bed nets (ITNs), as well as installment of mosquito screens in houses, may further reduce indoor transmission. For outdoor transmission, the use of community-centric approaches to reduce or eliminate larval sources within and surrounding the village through the guidance of locally stationed entomologists, along with Social and Behavior Change mediated health education towards the local adoption of mosquito protection tools during outdoor activities, may reduce malaria transmission.

Comparison of different trapping methods to collect malaria vectors indoors and outdoors in western Kenya.

BACKGROUND: Vector surveillance is among the World Health Organization global vector control response (2017-2030) pillars. Human landing catches are a gold standard but difficult to implement and potentially expose collectors to malaria infection. Other methods like light traps, pyrethrum spray catches and aspiration are less expensive and less risky to collectors. METHODS: Three mosquito sampling methods (UV light traps, CDC light traps and Prokopack aspiration) were evaluated against human landing catches (HLC) in two villages of Rarieda sub-county, Siaya County, Kenya. UV-LTs, CDC-LTs and HLCs were conducted hourly between 17:00 and 07:00. Aspiration was done indoors and outdoors between 07:00 and 11:00 a.m. Analyses of mosquito densities, species abundance and sporozoite infectivity were performed across all sampling methods. Species identification PCR and ELISAs were done for Anopheles gambiae and Anopheles funestus complexes and data analysis was done in R. RESULTS: Anopheles mosquitoes sampled from 608 trapping efforts were 5,370 constituting 70.3% Anopheles funestus sensu lato (s.l.), 19.7% Anopheles coustani and 7.2% An. gambiae s.l. 93.8% of An. funestus s.l. were An. funestus sensu stricto (s.s.) and 97.8% of An. gambiae s.l. were Anopheles arabiensis. Only An. funestus were sporozoite positive with 3.1% infection prevalence. Indoors, aspiration captured higher An. funestus (mean = 6.74; RR = 8.83, P < 0.001) then UV-LT (mean = 3.70; RR = 3.97, P < 0.001) and CDC-LT (mean = 1.74; RR = 1.89, P = 0.03) compared to HLC. UV-LT and CDC-LT indoors captured averagely 0.18 An. arabiensis RR = 5.75, P = 0.028 and RR = 5.87, P = 0.028 respectively. Outdoors, UV-LT collected significantly higher Anopheles mosquitoes compared to HLC (An. funestus: RR = 5.18, P < 0.001; An. arabiensis: RR = 15.64, P = 0.009; An. coustani: RR = 11.65, P < 0.001). Anopheles funestus hourly biting indoors in UV-LT and CDC-LT indicated different peaks compared to HLC. CONCLUSIONS: Anopheles funestus remains the predominant mosquito species. More mosquitoes were collected using aspiration, CDC-LTs and UV-LTs indoors and UV-LTs and CD-LTs outdoors compared to HLCs. UV-LTs collected more mosquitoes than CDC-LTs. The varied trends observed at different times of the night suggest that these methods collect mosquitoes with diverse activities and care must be taken when interpreting the results.

Evaluation of the effectiveness of the human-baited double net traps and BG traps compared with the human landing catches for collecting outdoor Aedes albopictus in China.

Dengue fever is one of the biggest threats to public health in China, causing huge disease burden and economic loss. Aedes-mosquito surveillance could be a cornerstone for predicting the risk of Aedes-borne diseases and evaluating the effect of vector management during diseases outbreaks. The human landing catch (HLC) method is regarded as the "gold standard" for catching Aedes mosquitoes, but it potentially exposes field professionals to vectors of known or unknown pathogens. Human-baited double net (HDN) was recommended to replace HLC for emergency monitoring in China when Aedes-borne diseases break out, but it had been reported with low efficiency for capturing Aedes mosquitoes. In this study, we compared HLC with HDN and BG traps for field Aedes albopictus monitoring, with the aim of evaluating the effectiveness of HDN replacing HLC and finding an effective and safe alternative to the HLC for monitoring Aedes albopictus. Six sites in Hangzhou, Shaoxing, and Yiwu, Zhejiang Province, China, were chosen to conduct outdoor HLC, HDN, and BG trap catches from June to October 2021. The tests were performed 3 h apart: 8:30-9:30 AM, 16:30-17:30 PM, and 17:30-18:30 PM. A total of 2330 adult mosquitoes were collected, and Aedes albopictus was the most abundant species in all three catches with 848(98.95%), 559(97.39%) and 867 (96.44%) caught in HLC, HDN and BG traps respectively. Compared to HLC, HDN collected significantly less Ae. albopictus and Ae. albopictus females per trapping period (P < 0.001, P < 0.001), whereas no statistical differences were found between the HLC and BG trap (P = 0.970, P > 0.05). Statistically significant positive spatial correlations for Ae. albopictus sampling was found between HLC and HDN traps (r = 0.543, P < 0.001) and HLC and BG traps (r = 0.658, P < 0.001). In conclusion, both the BG trap and HDN have a significant positive spatial correlation with HLC, making them safer alternatives to HLC for Ae. albopictus monitoring in China. However, with better a sampling efficiency, being less labor intensive, and no human-baited attraction bias, the BG trap could be a better choice than the HDN trap.

Comparison of standard and modified human landing catching techniques for blackfly collection.

BACKGROUND: Human landing catches (HLCs) are required to collect blackflies for entomological evaluation to verify onchocerciasis elimination. However, there are ethical concerns regarding exposure of vector collectors to infectious blackflies and safer alternative methods are needed. This study evaluated a modified HLC technique where collectors wore coloured trousers (blue, black or blue-black), protecting them from bites during fly collection, and their performance was compared with the standard. METHODS: The study was conducted in Makouopsap, Cameroon, in the Massangam health district for 4 months. Four collector pairs-one 'standard' (bare-legged) and three modified-were placed 50 m apart along known breeding sites on the Mbam and Nja Rivers. Collections were performed from 07:00 to 17:00 h, 4 d/month. Hourly rates of flies caught were analysed using a negative binomial generalised linear model to explore associations between flies caught and collection techniques and seasons. RESULTS: Overall, 17 246 blackflies were caught. There was no significant statistical difference in the number of blackflies and parous flies caught between black trousers and the standard. Thus there is a strong indication that wearing black trousers is a viable non-inferior alternative to the standard HLC. CONCLUSIONS: Further studies are needed to confirm generalisability in different ecozones and transmission environments and among different blackfly species.

A semi-field evaluation in Thailand of the use of human landing catches (HLC) versus human-baited double net trap (HDN) for assessing the impact of a volatile pyrethroid spatial repellent and pyrethroid-treated clothing on Anopheles minimus landing.

BACKGROUND: The mosquito landing rate measured by human landing catches (HLC) is the conventional endpoint used to evaluate the impact of vector control interventions on human-vector exposure. Non-exposure based alternatives to the HLC are desirable to minimize the risk of accidental mosquito bites. One such alternative is the human-baited double net trap (HDN), but the estimated personal protection of interventions using the HDN has not been compared to the efficacy estimated using HLC. This semi-field study in Sai Yok District, Kanchanaburi Province, Thailand, evaluates the performance of the HLC and the HDN for estimating the effect on Anopheles minimus landing rates of two intervention types characterized by contrasting modes of action, a volatile pyrethroid spatial repellent (VSPR) and insecticide-treated clothing (ITC). METHODS: Two experiments to evaluate the protective efficacy of (1) a VPSR and (2) ITC, were performed. A block randomized cross-over design over 32 nights was carried out with both the HLC or HDN. Eight replicates per combination of collection method and intervention or control arm were conducted. For each replicate, 100 An. minimus were released and were collected for 6 h. The odds ratio (OR) of the released An. minimus mosquitoes landing in the intervention compared to the control arm was estimated using logistic regression, including collection method, treatment, and experimental day as fixed effects. RESULTS: For the VPSR, the protective efficacy was similar for the two methods: 99.3%, 95% CI (99.5-99.0) when measured by HLC, and 100% (100, Inf) when measured by HDN where no mosquitoes were caught (interaction test p = 0.99). For the ITC, the protective efficacy was 70% (60-77%) measured by HLC but there was no evidence of protection when measured by HDN [4% increase (15-27%)] (interaction test p < 0.001). CONCLUSIONS: Interactions between mosquitoes, bite prevention tools and the sampling method may impact the estimated intervention protective efficacy. Consequently, the sampling method must be considered when evaluating these interventions. The HDN is a valid alternative trapping method (relative to the HLC) for evaluating the impact of bite prevention methods that affect mosquito behaviour at a distance (e.g. VPSR), but not for interventions that operate through tarsal contact (e.g., ITC).

Composition of Anopheles species and bionomic characteristics over the peak malaria transmission season in Bandarban, Bangladesh.

BACKGROUND: Joint efforts by government and non-government organizations have helped to reduce malaria in Bangladesh and set the country on a clear path to eventual malaria elimination. However, achieving that goal would be challenging without a comprehensive understanding of vector bionomics. METHODS: Targeted capturing of Anopheles mosquitoes over a rainy season, utilizing specific sampling methods, including human landing catches (HLCs), CDC-light traps (CDC-LTs), and pyrethrum spray catches (PSCs) were aimed to characterize entomological drivers of transmission in four sites of Bandarban, Bangladesh. RESULTS: Molecular characterization of a subset of 4637 mosquitoes has demonstrated the presence of at least 17 species whose capture rates were representative of the rainy season. Species compositions and bionomic traits did not vary between sites with Anopheles maculatus having the highest landing rate by HLCs and Anopheles vagus having the highest capture rate with CDC-LTs. Interestingly, Anopheles species compositions and capture rates varied significantly (p < 0.05) for An. vagus, between HLCs and its often-used proxy-CDC-LTs- suggesting impacts on downstream analysis. CDC-LTs capture rates demonstrated differing compositions with indoor and outdoor biting rates. For example, Anopheles nigerrimus and Anopheles nivipes were more endophagic by HLCs and more exophagic by CDC-LTs. The use of a cow-baited CDC-LT also demonstrated significantly different results when compared to a human-baited CDC-LT considering the high degree of anthropophily in these species. The exception to both zoophily and indoor resting was An. vagus, which demonstrated both anthropophily and high resting rates indoors-pointing to this species being a possible primary vector at this site. CONCLUSION: A diverse Anopheles fauna in Bandarban has been confirmed through molecular methods, highlighting the potential impact of sampling techniques. Given the complexity of the local ecosystem, a better understanding of mosquito behaviour and ecology is required to achieve the goal of malaria elimination in Bangladesh.

The MTego trap: a potential tool for monitoring malaria and arbovirus vectors.

BACKGROUND: Odour-baited traps are useful for vector surveillance and control. However, most existing traps have shown inconsistent recapture rates across different mosquito species, necessitating the need for more effective and efficient traps. The MTego trap with integrated thermal stimuli has been developed as an alternative trap. This study was undertaken to determine and compare the efficacy of the MTego trap to that of the Biogents (BG) modular BG-Pro (BGP) trap for sampling different mosquito species in a semi-field system. METHODS: Fully balanced Latin square design experiments (no-choice and dual choice) were conducted in semi-field chambers using laboratory-reared female Anopheles gambiae sensu stricto, Anopheles funestus, Anopheles arabiensis, Culex quinquefasciatus and Aedes aegypti. There were 16 replicates, and 50 mosquitoes of each species were released in each chamber per replicate. The evaluated traps were as follows: the MTego trap baited with PM6 (MT-PM6), the MTego trap baited with BG-Lure (BGL) (MT-BGL), and the BGP trap baited with BG-Lure (BGP-BGL). RESULTS: In the no-choice test, the MT-BGL and BGP-BGL traps captured a similar proportion of An. gambiae (31% vs 29%, P-value = 0.519) and An. funestus (32% vs 33%, P = 0.520). The MT-PM6 and BGP-BGL traps showed no significant difference in capturing Ae. aegypti (33% vs 31%, P = 0.324). However, the BGP-BGL caught more An. arabiensis and Cx. quinquefasciatus mosquitoes than the other traps (P < 0.0001). In the dual-choice test of MT-PM6 vs BGP-BGL, similar proportions of An. funestus (25% vs 27%, P = 0.473) and Ae. aegypti (29% vs 25%, P = 0.264) were captured in the traps, while the BGP-BGL captured more An. gambiae, An. arabiensis and Cx. quinquefasciatus mosquitoes than the MT-PM6 (P < 0.0001). CONCLUSIONS: This study demonstrated that the MTego trap has potential as a tool that can be used interchangeably with the BGP trap for sampling anthropophilic mosquitoes including African malaria vectors An. gambiae and An. funestus and the principal arbovirus vector Ae. aegypti.

Vertical dispersal of Aedes albopictus within multi-story buildings in downtown Shanghai, China.

BACKGROUND: Shanghai has numerous high-rise apartment and office buildings, but the effects of these high-rise spaces on the vertical dispersal, oviposition and blood feeding behavior of Aedes albopictus are unknown. METHODS: In six multi-story building blocks in downtown Shanghai, 174 mosq-ovitraps (MOT) were placed both indoors and outdoors for Ae. albopictus collection at different vertical heights from the 1st to 6th floors and a terrace on the 8th floor. Collections were made for 4 months. The human landing catch (HLC) method for Ae. albopictus monitoring was also conducted on 6 consecutive days on six floors of two of the six buildings to study the feeding behavior of Ae. albopictus at different heights. RESULTS: Both MOTs and HLCs collected Ae. albopictus at all monitored heights. The vertical distribution, oviposition pattern and biting behavior varied significantly among the seven heights (1st-6th floors and 8th floor) (mosq-ovitrap index (MOI): X2 = 140.616, df = 6, P < 0.001; HLC: F (5, 138) = 15.111, P < 0.001). The MOI at low heights (1st + 2nd floors) was significantly higher than that at medium (3rd + 4th floor, P < 0.001) and high heights (5th + 6th floors, P < 0.001), and there was no significant difference in the MOI for the 3rd-6th floors. The outdoor MOIs were significantly higher than indoor MOIs at all heights (outdoor 23.09% vs. indoor 9.58%, X2 = 74.121, df = 1, P < 0.001). Aedes albopictus HLC density on the ground floor was significantly higher than that on all other heights (5.04 vs. 0.13, 0.29, 0.58, 0.79 and 1.50 per half hour, P < 0.05), while no difference was detected among the heights above the ground floor (P > 0.05). CONCLUSIONS: Aedes albopictus is more common near the ground level, but it can easily disperse to higher floors in the multi-story buildings of urban Shanghai. No significant differences in Ae. albopictus density were detected within the 3rd-6th floors using MOT or HLC. This suggests that Ae. albopictus might also disperse to areas above the 6th floor and seek hosts there. Aedes albopictus prefers to oviposit outdoors; however, Ae. albopictus was also able to inhabit, oviposit and engage in blood-feeding behavior indoors on different floors. The three-dimensional dispersal pattern of Ae. albopictus in urban areas could facilitate arbovirus transmission and increase the difficulty of dengue control.

Influence of meteorological and seasonal parameters on the activity of Culicoides paraensis (Diptera: Ceratopogonidae), an annoying anthropophilic biting midge and putative vector of Oropouche Virus in Rondônia, Brazilian Amazon.

Biting midges of the genus Culicoides are insects of proven medical and veterinary importance, because of their role in the transmission of viruses, bacteria, protozoa and nematodes. Culicoides paraensis has been considered the main vector of the Oropouche Virus (OROV) in the urban cycle of the disease in the neotropics. Due to the great abundance of Culicoides spp. in the State of Rondônia and its epidemiological history of OROV, we investigated the biting activity in humans, the abundance as a function of meteorological parameters and seasonality, and the detection of OROV. Entomological collections occurred in three municipalities from Brazilian State of Rondônia: Porto Velho, Ariquemes, and Ouro Preto do Oeste. GLMM's were used to determine if Culicoides spp. abundance was predicted by seasonal, diurnal, and meteorological factors. Total RNA was extracted from insects and viral RNA detection was performed using the S segment as the target region of OROV via RT-qPCR. In total, 7315 individuals were captured and identified as C. paraensis. In the dry season, 1488 individuals (24.5%) were recorded, 4591 (75.5%) in the rainy season, with peaks of biting activity between 4pm and 6pm. All variables showed a significative effect on the midge abundance. The rainy season, temperature between 30 °C and 32 °C and relative air humidity between 75% and 85% were the main predictive parameters for capturing the highest average number of insects. Our results confirm diurnal activity of C. paraensis and its greatest abundance in rainy periods. No sample was positive for the OROV, which could be explained by the virus absence in local human populations, C. paraensis as a minor vector species in the sampled localities, and probable low rate of infection of biting midges. Our findings on hourly and seasonal biting activities can provide support to intervention actions regarding vector control and surveillance of this species. This was the first study to collect and analyze biting midges in a region where human OROV cases had already been detected, but without previous information on entomovirological surveillance.

Human landing catches provide a useful measure of protective efficacy for the evaluation of volatile pyrethroid spatial repellents.

BACKGROUND: The human landing catch (HLC) method, in which human volunteers collect mosquitoes that land on them before they can bite, is used to quantify human exposure to mosquito vectors of disease. Comparing HLCs in the presence and absence of interventions such as repellents is often used to measure protective efficacy (PE). Some repellents have multiple actions, including feeding inhibition, whereby mosquitoes may be unable to bite even if they land on a host. A comparison was made between the PE of the volatile pyrethroid spatial repellent (VPSR) transfluthrin determined using a landing method (HLC) and a biting method (allowing the mosquitoes that landed to blood-feed) to evaluate whether HLC is a suitable method for the estimation of the personal PE of a VPSR. METHODS: A fully balanced, two-arm crossover design study was conducted using a 6 × 6 × 2-m netted cage within a semi-field system. Hessian strips (4 m × 0.1 m) treated with a 5-, 10-, 15-, or 20-g dose of transfluthrin were evaluated against a paired negative control for three strains of laboratory-reared Anopheles and Aedes aegypti mosquitoes. Six replicates were performed per dose using either the landing or the biting method. The number of recaptured mosquitoes was analysed by negative binomial regression, and the PEs calculated using the two methods were compared by Bland-Altman plots. RESULTS: For Anopheles, fewer mosquitoes blood-fed in the biting arm than landed in the landing arm (incidence rate ratio = 0.87, 95% confidence interval 0.81-0.93, P < 0.001). For Ae. aegypti, biting was overestimated by around 37% with the landing method (incidence rate ratio = 0.63, 95% confidence interval 0.57-0.70, P = 0.001). However, the PEs calculated for each method were in close agreement when tested by the Bland Altman plot. CONCLUSIONS: The HLC method led to underestimation of mosquito feeding inhibition as a mode of action of transfluthrin, and there were species- and dose-dependent differences in the relationship between landing and biting. However, the estimated PEs were similar between the two methods. The results of this study indicate that HLC can be used as a proxy for personal PE for the evaluation of a VPSR, especially when the difficulties associated with enumerating blood-fed mosquitoes in a field setting are taken into consideration.

The effect of transfluthrin-treated jute and cotton emanator vests on human landing and fecundity of Anopheles minimus in Thailand.

Complementary approaches to tackle outdoor and early evening biting mosquitoes are urgently required. Transfluthrin (TFT) is a volatile pyrethroid capable of altering mosquito feeding behavior. This study investigated the efficacy of TFT-treated jute (TFT-J) and cotton (TFT-C) fabrics on human landing activity, feeding and fecundity of Anopheles minimus in Thailand. Jute and cotton fabrics each measuring 1024 cm2 were impregnated with 7.34 × 10-4 g/cm2 TFT (20%, w/v), and evaluated in a semi-field screen house system. Two collectors, wearing an untreated control or TFT-treated vests, conducted human-landing collections of released 100 laboratory-reared adult females of An. minimus from 18:00-00:00 h for 16 consecutive nights. Recaptured mosquitoes were given a blood meal for 30 min. with a membrane feeding system for assessment of blood feeding and fecundity. TFT-J, relative to control, significantly reduced human landings (Odds Ratio (OR) =0.27 (95% Confidence Interval (CI) [0.10-0.74], p = 0.011)), however no significant reduction was observed for TFT-C (OR=0.67 [95% CI 0.24-1.82], p = 0.43). Blood feeding was significantly lower among mosquitoes exposed to TFT-J (12.45% [95% CI, 2.04-22.85], p = 0.029) and TFT-C (13% [95% CI, 0.99-26.84], p = 0.016) relative to control. Impregnated fabrics had no effect on the mean number of egg oviposition. However, egg hatchability was reduced in TFT-J (49.5% [95% CI, 21.74-77.26], p = 0.029) and TFT-C (40.2% [95% CI, 17.21-63.19], p = 0.008) relative to control. TFT-J significantly reduced the landing, blood feeding, and fertility of An. minimus. Further studies are needed to evaluate different treatment methods on fabrics and their incorporation in integrated mosquito management.

Human biting mosquitoes and implications for West Nile virus transmission.

BACKGROUND: West Nile virus (WNV), primarily vectored by mosquitoes of the genus Culex, is the most important mosquito-borne pathogen in North America, having infected thousands of humans and countless wildlife since its arrival in the USA in 1999. In locations with dedicated mosquito control programs, surveillance methods often rely on frequent testing of mosquitoes collected in a network of gravid traps (GTs) and CO2-baited light traps (LTs). Traps specifically targeting oviposition-seeking (e.g. GTs) and host-seeking (e.g. LTs) mosquitoes are vulnerable to trap bias, and captured specimens are often damaged, making morphological identification difficult. METHODS: This study leverages an alternative mosquito collection method, the human landing catch (HLC), as a means to compare sampling of potential WNV vectors to traditional trapping methods. Human collectors exposed one limb for 15 min at crepuscular periods (5:00-8:30 am and 6:00-9:30 pm daily, the time when Culex species are most actively host-seeking) at each of 55 study sites in suburban Chicago, Illinois, for two summers (2018 and 2019). RESULTS: A total of 223 human-seeking mosquitoes were caught by HLC, of which 46 (20.6%) were mosquitoes of genus Culex. Of these 46 collected Culex specimens, 34 (73.9%) were Cx. salinarius, a potential WNV vector species not thought to be highly abundant in upper Midwest USA. Per trapping effort, GTs and LTs collected > 7.5-fold the number of individual Culex specimens than HLC efforts. CONCLUSIONS: The less commonly used HLC method provides important insight into the complement of human-biting mosquitoes in a region with consistent WNV epidemics. This study underscores the value of the HLC collection method as a complementary tool for surveillance to aid in WNV vector species characterization. However, given the added risk to the collector, novel mitigation methods or alternative approaches must be explored to incorporate HLC collections safely and strategically into control programs.

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

BACKGROUND: Methods 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. METHODS: Between 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. RESULTS: Overall, 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%). CONCLUSION: In 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.

Outdoor and early hour human biting activities of malaria mosquitoes and the suitability of clay pot for outdoor resting mosquito collection in malaria endemic villages of southern Rift Valley, Ethiopia.

Background: Sampling adult Anopheles mosquitoes is important for assessing vector density, estimating the sporozoite infection rate, and quantifying the impact of vector control interventions. The objective of this study was to assess the Anopheles mosquito species composition, and their outdoor and indoor biting activities, and to evaluate the suitability of clay pots for indoor and outdoor resting mosquito collections. Methods: Two malaria-endemic villages in the Gamo zone were purposely selected. Forty clay pots were deployed for outdoor resting mosquitoes sampling and another forty for indoor resting sampling. Twenty pit shelters were constructed for outdoor resting mosquito collection. The human landing catch (HLC) technique was employed to collect indoor and outdoor host-seeking mosquitoes in two households in each village. Morphological identification of the Anopheles mosquito was done using an identification key. Enzyme-linked immunosorbent assay technique was used for blood meal origin and circumsporozoite proteins (CSP) test. Speciation of An. gambiae complex was done using polymerase chain reaction. A Chi-square test was used to compare the effectiveness of clay pot and pit shelters for outdoor resting sampling. Results: A total of 904 female Anopheles mosquitoes comprising An. gambiae complex, An. pharoensis, An. tenebrosus, An. dencalicus and An. demelloni were sampled. The majority (64%) of them were sampled by the HLC technique. There was a slight difference between the outdoor clay pot (19%) and pit shelter (17%) collection. No Anopheles mosquitoes were collected indoor using clay pots. All mosquitoes were tested for CSPs, but none of them were found to be positive. Anopheles mosquitoes were tending to bite humans outdoor than indoors, and their peak biting hours was 10-11 pm. The human blood meal index of Anopheles mosquitoes was 0.07 from pit shelters and it was 0.04 from clay pots. The bovine blood meal index was 0.45 for mosquitoes from both pit shelters and clay pot collections. Conclusion: Anopheles arabiensis was the predominant species and it was tending to bite cattle more than humans. Clay pot could be suitable for outdoor resting mosquito collection, but not for indoor resting species.

Comparison of Vector Trapping Methods for Outdoor Biting Malaria Vector Surveillance in Thailand and Vietnam.

The performances of the human-baited double net trap (HDNT) and the human-baited host decoy trap (HDT) methods were compared against the outdoor human landing catch (OHLC) method in Thailand and Vietnam. Two study sites were selected in each country: a rural village and a nearby forest setting. The three outdoor trap methods were rotated nightly between three set trapping positions, in a pre-assigned Latin square design. Volunteers were rotated following the trap rotation to avoid bias. The greatest number of adult mosquitoes was collected from the forest sites in both countries, showing Anopheles minimus (s.s.) Theobald (96.54%) and Anopheles dirus (s.s.) Peyton & Harrison (25.71%) as the primary malaria vectors in Thailand and Vietnam, respectively. At the Thai forest site, OHLC collected significantly more anopheline mosquitoes per trap night than HDNT and HDT, with mean ± standard error values of 14.17 ± 4.42, 4.83 ± 1.56, and 4.44 ± 1.45, respectively, whilst HDNT and HDT were significantly less productive at 0.34 times and 0.31 times, respectively, than OHLC in capturing anopheline mosquitoes. However, there were no significant differences among the three methods of trapping malaria vectors for the village site. At the Vietnamese forest site, HDNT achieved the highest performance in collecting Anopheline mosquitoes at 1.54 times compared to OHLC, but there was no significant difference between the two traps. The results suggested HDNT could be a possible alternative trap to OHLC in this area. Although HDT was less efficient at attracting Anopheline mosquitoes, it was highly efficient at trapping culicine mosquitoes.

Anopheline diversity in urban and peri-urban malaria foci: comparison between alternative traps and seasonal effects in a city in the Western Brazilian Amazon.

BACKGROUND: Continuous vector surveillance and sustainable interventions are mandatory in order to prevent anopheline proliferation (or spread to new areas) and interrupt malaria transmission. Anopheline abundance and richness were evaluated in urban and peri-urban malaria foci at a medium-sized city in the Brazilian Amazon, comparing the protected human landing catch technique (PHLC) and alternative sampling methods over different seasonal periods. Additional information was assessed for female feeding behaviour and faunal composition. METHODS: Anophelines were sampled bimonthly in four urban and peri-urban sites in the city of Porto Velho, state of Rondônia, Brazil. The average number of captured mosquitoes was compared between an PHLC (gold standard), a tent trap (Gazetrap), and a barrier screen by means of generalized linear mixed models (GLMM), which also included season and environment (peri-urban/urban) as predictors. RESULTS: Overall, 2962 Anopheles individuals belonging to 12 species and one complex were caught; Anopheles darlingi represented 86% of the individuals. More mosquitoes were captured in the peri-urban setting, and the urban setting was more diverse. The model estimates that significantly more anophelines were collected by PHLC than by the Screen method, and Gazetrap captured fewer individuals. However, the Screen technique yielded more blood-engorged females. The peak hours of biting activity were from 6 to 7 p.m. in urban areas and from 7 to 8 p.m. in peri-urban areas. CONCLUSIONS: Although peri-urban settings presented a greater abundance of anophelines, Shannon and Simpson diversities were higher in urban sites. Each technique proved to be useful, depending on the purpose: PHLC was more effective in capturing the highest anopheline densities, Gazetrap caught the greatest number of species, and the barrier screen technique captured more engorged individuals. There was no seasonal effect on Anopheles assemblage structure; however, a more diverse fauna was caught in the transitional season. Biting activity was more intense from 6 to 8 p.m., with a predominance of An. darlingi.

Less is more: repellent-treated fabric strips as a substitute for full screening of open eave gaps for indoor and outdoor protection from malaria mosquito bites.

BACKGROUND: Providing protection from malaria vector bites, both indoors and outdoors, is crucial to curbing malaria parasite transmission. Screening of house entry points, especially with incorporated insecticides, confers significant protection but remains a costly and labour-intensive application. Use of spatial repellents has shown promise in creating areas of protection in peri-domestic areas. METHODS: This study aimed at comparing the protection provided by transfluthrin-treated and untreated complete screens over open eave gaps with incomplete transfluthrin-treated eave strips as a potential replacement for a full screen. Human landing catches were implemented independently inside and outside an experimental hut under controlled semi-field conditions, with insectary-reared Anopheles arabiensis mosquitoes. RESULTS: The odds of a female mosquito finding a human volunteer indoors and attempting to bite were similar whether the eaves were completely open or there was an untreated fabric strip fixed around the eaves. However, when the eave gap was completely screened without insecticide, the odds of receiving a bite indoors were reduced by 70% (OR 0.30, 95% CI 0.20-0.47). Adding transfluthrin to the full screen, further increased the protection indoors, with the odds of receiving a bite reduced by 92% (0.08, 95% CI 0.04-0.16) compared to the untreated screen. Importantly, the same protection was conferred when only a narrow transfluthrin-treated fabric strip was loosely fixed around the eave gap (OR 0.07, 95% CI 0.04-0.13). The impact of the transfluthrin treatment on outdoor biting was correlated with evening temperatures during the experiments. At lower evening temperatures, a transfluthrin-treated, complete screen provided moderate and variable protection from bites (OR 0.62, 95% CI 0.37-1.03), whilst at higher evening temperatures the odds of receiving a bite outdoors was over four times lower in the presence of transfluthrin, on either a full screen (OR 0.22 95% 0.12-0.38) or a fabric strip (OR 0.25, 95% 0.15-0.42), than when no treatment was present. CONCLUSION: The findings suggest that transfluthrin-treated fabric strips can provide a substitute for complete eave screens. They are a simple, easy-to-handle tool for protecting people from malaria mosquito bites indoors and potentially around the house in climatic areas where evening and night-time temperatures are relatively high.

Blood feeding habits of mosquitoes: hardly a bite in South America.

Mosquito blood feeding plays a key role in epidemiology. Despite its importance and large number of studies worldwide, less attention has been paid in South America. We summarized some general concepts and methodological issues related to the study of mosquito blood feeding habits, and compiled and analyzed all published information regarding the subject in the continent until 2020. Available literature comprised 152 scientific studies, that pursued different approaches: human landing catches (102 studies), baited trap (19), and blood meal analyses of collected specimens (38). Among the latter, 23 used serological and 15 molecular techniques. Species most frequently studied were those incriminated in malaria transmission, whereas relevant vectors such as Aedes aegypti, Ae. albopictus, and Haemagogus janthinomys were surprisingly neglected. Brazil was the leading country both in number of works and species studied. For over 70% of the species and three out of 13 South American countries there is no single information on mosquito blood feeding habits. Data from baited traps included 143 mosquito species, 83.9% of which were attracted to humans, either exclusively (10.5%) or in combination with other vertebrates (73.4%). Host blood identification of field collected specimens provided data on 102 mosquito species, and 60.8% of these fed on humans (55.9% combined with other vertebrates). Only 17 of the 73 species assessed by both methods yielded similar feeding patterns. Finally, supplementary tables are provided in a comprehensive summary of all information available and information gaps are highlighted for future research in the continent.

Larvae ecology and adult activity of Aedes mariae (Diptera: Culicidae) in a touristic rock-pool area of the Balearic Islands (Western Mediterranean).

Mosquitoes are vectors of several diseases of medical concern such as malaria or dengue and can also negatively affect tourism and the life-quality of the neighbourhood. The species Aedes mariae (Sergent and Sergent, 1903) is a poorly studied mosquito that breeds in rock-pools of the Mediterranean coast. General Linear Mixed Models (GLMM) were used to determine drivers affecting the presence and abundance of this species. Abiotic and biotic factors were recorded in rock-pools with the presence of Ae. mariae sub-adults across a supralittoral area of Majorca Island (Balearic Islands, Spain) from July 2018 to June 2019. We tested how abiotic factors affected the presence of larvae, while the biotic factors were used to check their effect on larvae abundance. human landing collection was also conducted to assess the adult activity of this species. Valuable data were recorded to improve our knowledge about the bioecology of Ae. mariae in a touristic area of the island of Majorca. Salinity and pH were the most explanatory variables for the presence of Ae. mariae larvae. The presence of Posidonia oceanica (L.) Delile 1813 leaves negatively affected the abundance of Ae. mariae larvae while the presence of other fauna enhanced it. Adult females of Ae. mariae were active for 26 min after sunset in June and its host-seeking activity decreased during autumn months. Control methods against this species should be focussed on rock-pools and planning treatments according to tides, waves and precipitation.

Semi-field evaluation of the exposure-free mosquito electrocuting trap and BG-Sentinel trap as an alternative to the human landing catch for measuring the efficacy of transfluthrin emanators against Aedes aegypti.

BACKGROUND: The human landing catch (HLC) measures human exposure to mosquito bites and evaluates the efficacy of vector control tools. However, it may expose volunteers to potentially infected mosquitoes. The mosquito electrocuting trap (MET) and BG-Sentinel traps (BGS) represent alternative, exposure-free methods for sampling host-seeking mosquitoes. This study investigates whether these methods can be effectively used as alternatives to HLC for measuring the efficacy of transfluthrin emanator against Aedes aegypti. METHODS: The protective efficacy (PE) of freestanding passive transfluthrin emanators (FTPEs), measured by HLC, MET and BGS, was compared in no-choice and choice tests. The collection methods were conducted 2 m from an experimental hut with FTPEs positioned at 3 m on either side of them. For the choice experiment, a competitor HLC was included 10 m from the first collection point. One hundred laboratory-reared Ae. aegypti mosquitoes were released and collected for 3 consecutive h. RESULTS: In the no-choice test, each method measured similar PE: HLC: 66% (95% confidence interval [CI]: 50-82), MET: 55% (95% CI: 48-63) and BGS: 64% (95% CI: 54-73). The proportion of mosquitoes recaptured was consistent between methods (20-24%) in treatment and varied (47-71%) in the control. However, in choice tests, the PE measured by each method varied: HLC: 37% (95% CI: 25-50%), MET: 76% (95% CI: 61-92) and BGS trap: 0% (95% CI: 0-100). Recaptured mosquitoes were no longer consistent between methods in treatment (2-26%) and remained variable in the control (7-42%). FTPE provided 50% PE to the second HLC 10 m away. In the control, the MET and the BGS were less efficacious in collecting mosquitoes in the presence of a second HLC. CONCLUSIONS: Measuring the PE in isolation was fairly consistent for HLC, MET and BGS. Because HLC is not advisable, it is reasonable to use either MET or BGS as a proxy for HLC for testing volatile pyrethroid (VP) in areas of active arbovirus-endemic areas. The presence of a human host in close proximity invalidated the PE estimates from BGS and METs. Findings also indicated that transfluthrin can protect multiple people in the peridomestic area and that at short range mosquitoes select humans over the BGS.