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Understanding mosquito behaviours is vital for the development of insecticide-treated nets (ITNs), which have been successfully deployed in sub-Saharan Africa to reduce disease transmission, particularly malaria. However, rising insecticide resistance (IR) among mosquito populations, owing to genetic and behavioural changes, poses a significant challenge. We present a machine learning pipeline that successfully distinguishes between innate IR and insecticide-susceptible (IS) mosquito flight behaviours independent of insecticidal exposure by analysing trajectory data. Data-driven methods are introduced to accommodate common tracking system shortcomings that occur due to mosquito positions being occluded by the bednet or other objects. Trajectories, obtained from room-scale tracking of two IR and two IS strains around a human-baited, untreated bednet, were analysed using features such as velocity, acceleration, and geometric descriptors. Using these features, an XGBoost model achieved a balanced accuracy of 0.743 and a ROC AUC of 0.813 in classifying IR from IS mosquitoes. SHAP analysis helped decipher that IR mosquitoes tend to fly slower with more directed flight paths and lower variability than IS-traits that are likely a fitness advantage by enhancing their ability to respond more quickly to bloodmeal cues. This approach provides valuable insights based on flight behaviour that can reveal the action of interventions and insecticides on mosquito physiology., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Malaria remains a major public health threat in Burkina Faso, as in most sub-Saharan Africa countries. Malaria control relies mainly on long-lasting insecticide-treated nets (LLINs) and indoor residual spraying. In Burkina Faso, an escalating of insecticide resistance has been observed over the last decades. This study aimed to investigate insecticide resistance and the underlying mechanisms in Anopheles gambiae complex in Ouagadougou. Anopheles gambiae s.l. larvae were collected from gutters and ponds, in Zogona, Tampouy and Tanghin, 3 localities in Ouagadougou from July to October 2018. The larvae were reared in the laboratory to adults stage and susceptibility profile to pyrethroid, carbamate, and organophosphate insecticides was assessed using WHO tube assays. Mosquito species and mutations linked with insecticide resistance, were identified through PCR. More than 95% of the collected An. gambiae s.l. were An. arabiensis. An. arabiensis displayed high resistance to permethrin and deltamethrin, with mortalities below 30%, but was fully susceptible to bendiocarb, fenitrothion, and malathion. A high-frequency of the pyrethroid resistance-associated kdr mutation 1014F (0.81) was recorded, while the frequency of 1014S mutation (0.18) was lower. However, the carbamate and organophosphate-associated Ace-1 119S mutation was not detected. Localities and breeding site type appear to influence pyrethroid resistance in the An. arabiensis population of Ouagadougou. The high resistance to pyrethroids in An. arabiensis of urban Ouagadougou is underpinned, at least in part by high-frequency kdr mutations. This result supports the switch to next-generation LLINs, in well-established pyrethroid resistance zones of Burkina Faso including Ouagadougout., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Resistance to insecticides is spreading among populations of Aedes aegypti, the primary vector of important human arboviruses. The escalating insecticide resistance poses a significant threat to dengue vector control, with an expanding number of countries affected by the disease. To gain a deeper insight into the evolution of insecticide resistance, it is essential to have longitudinal surveillance results, which are currently lacking, particularly from African Ae. aegypti populations. Here we report on three-years of surveillance of Ae. aegypti susceptibility to insecticide resistance phenotypes and associated kdr mutations in Burkina Faso, a country with regular dengue outbreaks., Methods: Ae. aegypti susceptibility to insecticides and the V410L, V1016I, and F1534C kdr target site mutations linked to pyrethroid insecticide resistance were monitored in Ouagadougou from 2016 to 2018. Larvae were collected from artificial containers at two sites and reared to adulthood in an insectary. Bioassays were conducted on female adults, along with a laboratory-susceptible strain, following standard WHO protocols. Allele-specific PCR genotyping assays were utilized to identify the V410L, V1016I, and F1534C kdr pyrethroid target site mutations., Results: Bioassays revealed a high level of resistance to permethrin and deltamethrin that progressively increased over the three-year period in both localities. The 1534C mutation was nearly fixed throughout the three years at each locality, and while the closely-related 410L and 1016I mutations did not vary between localities, their frequency notably increased from 2016 to 2018. Interestingly, Ae. aegypti populations in both areas remained susceptible to bendiocarb, fenitrothion, and malathion. Modelling the mortality data further confirmed the escalating resistance trend over the years and emphasized the significant role played by the three kdr mutations in conferring resistance to pyrethroids., Conclusion: Mortality rates indicate that Ae. aegypti populations from Ouagadougou are becoming increasingly resistant to pyrethroid insecticides, likely due to an increase in the frequencies of the 410L and 1016I kdr mutations. Organophosphate insecticides are likely to be better alternative options for control., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Yaméogo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Recent dengue outbreaks have occurred in Ouagadougou and Bobo-Dioulasso, the two major cities of Burkina Faso. Dengue is a viral disease transmitted primarily by Aedes aegypti, a highly anthropophilic mosquito that thrives in human-transformed environments and breeds predominantly in artificial containers. In 2018, we investigated the resting and blood-feeding habits of Ae. aegypti in urban settings of Ouagadougou. In a 3-month cross-sectional study starting in August 2018, indoors and outdoors resting adult mosquitoes were collected using Prokopack aspirators in three health districts (HD). All mosquitoes were morphologically identified, and DNA was extracted from blood-fed Ae. aegypti females. A multiplex polymerase chain reaction with specific primers was used to identify the origin of the blood meal. A total of 4,256 adult Ae. aegypti mosquitoes, including 1,908 females, were collected. A preference for exophily was recorded in Bogodogo and Nongremassom, although an unexpectedly higher proportion of blood-fed females were found indoors than outdoors. Respectively, 96.09%, 91.03%, and 95.54% of the blood meals successfully analyzed in Baskuy, Bogodogo, and Nongremassom were from a single human host, with the remainder from domestic mammals as single or multiple hosts. Modeling total Ae. aegypti and blood-fed female counts showed that among other predictors, human density, outdoor environment, and house type affect their total densities. Our study revealed an exophilic tendency as well as a pronounced anthropophilic preference of Ae. aegypti adults, critical findings to consider when planning accurate entomological surveillance and effective interventions against Ae. aegypti in urban settings.

Background: The widespread use of insecticide-treated nets (ITNs) has significantly contributed to the reduction in malaria cases and deaths observed across Africa. Unfortunately, this control strategy is threatened by the rapid spread of pyrethroid resistance in malaria vectors. Dual-active-ingredient insecticidal nets are now available to mitigate the impact of pyrethroid resistance. To facilitate evidence-based decisions regarding product selection in specific use settings, data are needed on the efficacy of these different nets against local mosquito populations., Methods: Two experimental hut trials were performed in Za-Kpota, southern Benin in 2021 to evaluate the performance of Interceptor G2 (BASF), Royal Guard (Disease Control Technologies) and PermaNet 3.0 (Vestergaard Frandsen), all dual-active-ingredient bednets, in comparison to untreated or standard pyrethroid-treated bednets, against free-flying wild Anopheles gambiae mosquitoes. The performance of some of these next-generation nets was compared to the same type of nets that have been in use for up to 2 years. Mosquitoes collected in the huts were followed up after exposure to assess the sublethal effects of treatments on certain life-history traits., Results: The predominant species in the study site was Anopheles gambiae sensu stricto (An. gambiae s.s.). Both Anopheles coluzzii and An. gambiae s.s. were resistant to pyrethroids (deltamethrin susceptibility was restored by piperonyl butoxide pre-exposure). In the experimental hut trials, the highest blood-feeding inhibition (5.56%) was recorded for the Royal Guard net, relative to the standard PermaNet 2.0 net (44.44% inhibition). The highest 72-h mortality rate (90.11%) was recorded for the Interceptor G2 net compared to the PermaNet 2.0 net (56.04%). After exposure, the risk of death of An. gambiae sensu lato (An. gambiae s.l.) was 6.5-fold higher with the Interceptor G2 net and 4.4-fold higher with the PermaNet 3.0 net compared to the respective untreated net. Lower mosquito mortality was recorded with an aged Interceptor G2 net compared to a new Interceptor G2 net. Oviposition rates were lower in mosquitoes collected from huts containing ITNs compared to those of untreated controls. None of the mosquitoes collected from huts equipped with Royal Guard nets laid any eggs., Conclusions: The Royal Guard and Interceptor G2 nets showed a potential to significantly improve the control of malaria-transmitting vectors. However, the PermaNet 3.0 net remains effective in pyrethroid-resistant areas., (© 2024. The Author(s).)

Background: Ae. aegypti is the vector of important μ arboviruses, including dengue, Zika, chikungunya and yellow fever. Despite not being specifically targeted by insecticide-based control programs in West Africa, resistance to insecticides in Ae. aegypti has been reported in countries within this region. In this study, we investigated the status and mechanisms of Ae. aegypti resistance in Niamey, the capital of Niger. This research aims to provide baseline data necessary for arbovirus outbreak prevention and preparedness in the country., Methods: Ovitraps were used to collect Ae. aegypti eggs, which were subsequently hatched in the insectary for bioassay tests. The hatched larvae were then reared to 3-5-day-old adults for WHO tube and CDC bottle bioassays, including synergist tests. The kdr mutations F1534C, V1016I, and V410L were genotyped using allele-specific PCR and TaqMan qPCR methods., Results: Ae. aegypti from Niamey exhibited moderate resistance to pyrethroids but susceptibility to organophosphates and carbamates. The kdr mutations, F1534C, V1016I and V410L were detected with the resistant tri-locus haplotype 1534C+1016L+410L associated with both permethrin and deltamethrin resistance. Whereas the homozygote tri-locus resistant genotype 1534CC+1016LL+410LL was linked only to permethrin resistance. The involvement of oxidase and esterase enzymes in resistance mechanisms was suggested by partial restoration of mosquitoes' susceptibility to pyrethroids in synergist bioassays., Conclusion: This study is the first report of Ae. aegypti resistance to pyrethroid insecticides in Niamey. The resistance is underpinned by target site mutations and potentially involves metabolic enzymes. The observed resistance to pyrethroids coupled with susceptibility to other insecticides, provides data to support evidence-based decision-making for Ae. aegypti control in Niger., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Maiga et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Insecticide-treated nets (ITNs) are the most effective method for malaria prevention in Africa. Using near-infrared video tracking in a laboratory environment, we recorded and assessed bednet entry and exit by a northern Tanzanian population of Anopheles arabiensis at a human-occupied untreated net and a PermaNet® 2.0 ITN. Both had 12 holes, each 10 cm in diameter, punctured at specific locations, and the ITN was washed 20 times to further simulate the wear and tear of ageing. Washing reduced the insecticide content of ITNs by 61%, which then showed similar rates to the untreated nets for net entry (39% entered untreated net and 41% entered ITN; p = 0.84) and exit (37% and 43%, respectively; p = 0.67). Regardless of treatment, approximately 40% of mosquitoes entered nets within 20 s of first appearing in the field of view and reached the volunteer's skin within 5 s of entering the net. Mortality rates post-exposure were significantly higher ( p = 0.048) at ITNs (26.6%; 95% CI 13.4%-39.7%) than at untreated controls (6.4%; 95% CI 1.8%-14.6%). The washed and aged ITN provided little additional personal protection for the sleeper over an untreated net. Simple adjustments to materials and design that could extend the effective lifespan of ITNs are discussed., Competing Interests: We declare we have no competing interests., (© 2024 The Authors.)

Barrier bednets (BBnets), regular bednets with a vertical insecticidal panel to target mosquitoes above the bednet roof, where they are most active, have the potential to improve existing Insecticidal Treated Bednets (ITNs), by reducing the quantity of insecticide required per net, reducing the toxic risks to those using the net, potentially increasing insecticide choice. We evaluated the performance of PermaNet 3.0 (P3) and untreated (Ut) bed nets with and without pyrethroid and piperonyl butoxide roof barriers in killing pyrethroid-resistant and susceptible Anopheles gambiae, simultaneously video-recording mosquito flight tracks. Bioassay results showed that treated roof barriers, particularly the longitudinal P3 barrier (P3L) could be an effective addition to a bed net: P3 + P3L were consistently significantly more effective than the reference P3 bednet while performance of untreated nets could be raised to equal that of the reference P3 following the addition of a P3 barrier. The BBnet's potential to augment existing bednets and enhance their performance is considered., (© 2023. The Author(s).)

Background: Insecticide-treated nets (ITNs) using pyrethroids have been the main vector control tools deployed in malaria endemic countries and are responsible for the dramatic reduction in African malaria cases in the early 2000s. The World Health Organization (WHO) cone test was designed to assess the rapid toxicity effects of pyrethroid exposure on mosquito vectors but has yielded no insights beyond 60-min knockdown and 24-h mortality. As dual-active-ingredient (AI) ITNs become more widespread, bioassays that can provide realistic assessment of single- and dual-treated ITNs (i.e. nets with more than one active ingredient) are urgently needed., Methods: We present an augmentation of the cone test that enables accurate quantification of vector behavioural responses (specifically movement, spatial and temporal occupancy) to ITNs using video recording and bespoke software that uses background segmentation methods to detect spatial changes in the movement of mosquitoes within the cone. Four strains of Anopheles gambiae sensu lato (s.l.) were exposed to four ITNs (PermaNet 2.0, PermaNet 3.0, Olyset Net, Interceptor G2) and untreated nets in these modified cone tests. Life history data (post-exposure blood-feeding, blood meal weight, longevity) for individual mosquitoes were recorded., Results: All mosquitoes responded to the presence of ITNs, spending from 1.48 to 3.67 times more time in the upper region of the cone, depending on the ITN type. Of all ITNs, PermaNet 2.0 provoked the smallest change in behavioural response. Activity in the cone influenced observed post-exposure longevity, and in resistant strains exposed to Interceptor G2, the higher the activity, the greater the risk of dying, as long as the proportion of activity at the net surface was less than 50%. All ITNs inhibited blood-feeding, and smaller blood meals were taken when mosquitoes fed., Conclusions: The additional mosquito behaviour data obtained by using this modification to the WHO cone test provides unique insight into the innate responses of different mosquito strains on untreated nets and the entomological mode of action of ITNs, important evidence when evaluating ITN characteristics., (© 2023. The Author(s).)

Background: The human biting rate (MBR) and entomological inoculation rate (EIR) are common parameters routinely used to measure the risk of malaria transmission. Both parameters can be estimated using human landing catches (HLC). Although it is considered the gold-standard, HLC puts collectors at higher risk of infection with mosquito-transmitted pathogens. Methods: A novel exposure-free host-seeking mosquito electrocution trap, the Shockwè trap (SHK), was developed and its efficiency for monitoring mosquito community composition and abundance was compared with human landing catches (HLC) as the gold-standard. Field experiments were performed in Massavasse village, southern Mozambique. Simultaneous indoor and outdoor collections of nocturnal host-seeking mosquitoes were carried out using the SHK and HLC methods. The relative sampling efficiency of SHK was estimated as the ratio of the numbers of mosquitoes caught in SHK compared HLC. Proportionality and density-dependence between SHK and HLC catches were estimated by mean of Bayesian regression approaches. Results: A total of 69,758 and 27,359 host-seeking mosquitoes comprising nineteen species and four genera, were collected by HLC and SHK respectively. In general, SHK and HLC sampled similar numbers of mosquito species, with the exceptions of the least common species Aedes sudanensis , Ae. subargenteus , and Coquillettidia versicolor that were caught only by HLC. The relative sampling efficiency and proportionality between SHK and matched HLC catches varied greatly between species and collection site. However, all mosquitoes collected by SHK were unfed, confirming the Shockwè trap design's performance and reliability as a successful mosquito exposure free sampling approach. Conclusions: Results demonstrate that SHK is a safe and reliable human-exposure free device for monitoring the occurrence of a wide range of mosquito, including major malaria and arboviruses vector species. However, improvements are needed to increase its sampling efficiency for less abundant mosquito species., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Kampango A et al.)

Many mosquito vectors rest inside human habitations, a behavioral trait that is exploited for vector control by indoor residual spraying (IRS) of interior walls with insecticide. Although IRS and its refined version targeted IRS are very effective against Aedes aegypti , they are expensive and logistically challenging to deliver in densely populated urban areas where outbreaks of dengue and other arboviruses are the greatest challenge. In experiments in Recife, Brazil, we set out to quantify the indoor resting behavior of Ae. aegypti at a level beyond that previously reported. We found that significantly more Ae. aegypti males, unfed and fed females visited the base of walls (height 0-20 cm, corresponding to 12.3% of the total wall surface) more frequently than upper wall areas, with the difference more pronounced at higher temperatures. When the lowest 20 cm of the walls was treated with an appropriate insecticide and colored black, we recorded up to 85% cumulative mortality after 24-h exposure in the experimental room. The findings are significant because feasibly, householders could treat this small and accessible target zone manually, without the need for visits by costly IRS teams or equipment, reducing insecticide use and enabling communities to actively protect their own indoor environment., (© The Author(s) 2023. Published by Oxford University Press on behalf of National Academy of Sciences.)

The blood feeding performance of female mosquitoes directly impacts their ability to transmit malaria. Yet their host seeking and blood feeding behaviours in the presence of insecticide-treated nets (ITNs) are still poorly understood. This work explores how both insecticide resistant and susceptible Anopheles gambiae s.l. mosquitoes interact with pyrethroid nets (PermaNet 2.0 or Olyset net) or an untreated net (UTN) while attempting to blood feed on a human arm. Regardless of mosquito resistance status, the ITNs did not efficiently prevent host searching but reduced blood feeding success by 34.1 (29.31-38.95) %. The Permanet and Olyset net reduced to 227.5 (208.19-246.77) sec and 235.9 (214.03-257.74) sec the average blood feeding duration from 369.9 (342.78-397.04) sec with the UTN. The ingested blood volume was on average 22% lower for all mosquitoes exposed to insecticide. When feeding through ITNs, the blood volume flow rate of the susceptible strain increased by 35%, but no significant difference was found in the resistant strain. Thus, whilst the presence of the insecticide in ITNs reduced mosquito blood feeding success and blood volume, the mosquito's ability to respond by accelerating her rate of blood ingestion may further reduce the impact of ITNs on resistant mosquitoes., (© 2023. The Author(s).)

Background: Understanding the magnitude of human exposure to mosquito biting is fundamental to reduce pathogen transmission. Here we report on a study quantifying the levels of mosquitoes attacking humans throughout the night in a rural area of Southern Mozambique. Methods: Surveys were carried out in Massavasse village, southern Mozambique. The abundance and composition of host-seeking mosquito communities at night were assessed by human-landing catches (HLC) at one-hour intervals. Periods when people were located predominantly outdoors or indoors were used to estimate the amount of residents' exposure to mosquito bites in either location, to explore the potential impact a bed net could have had in reducing biting by each vector species. Results: A total of 69,758 host-seeking female mosquitoes comprising 23 species in four genera were collected. The exposure to biting by virtually all vector species was consistently high outdoors, typically at early evening and morning, with exception of An. gambiae s.l which was likely of biting a person with nearly same intensity indoors and outdoors throughout the night. Bed nets use could have reduced biting by An. gambiae s.l (dominated by An. arabiensis ), Ma. africana , Ma. uniformis , Cx. pipiens , Cx. antennatus , and Cx. poicilipes by 53%, 47%, 46%, 38%, 31%, and 28% respectively, compared to non-users. Conversely, a bed net user would have had little protection against An. pharoensis , An. ziemanni , An. tenebrosus , and Cx. tritaeniorhynchus biting exposures. Conclusions: This study showed that Massavasse residents were exposed to high levels of outdoor biting by malaria and arbovirus vectors that abound in the village. The findings help to identify entomological drivers of persistent malaria transmission in Mozambique and identify a wide range of arbovirus vectors nocturnally active in rural areas, many with outbreak potential. The study highlights the need for a surveillance system for monitoring arboviral diseases vectors in Mozambique., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Kampango A et al.)

Background: The success of insecticide treated bed nets (ITNs) for malaria vector control in Africa relies on the behaviour of various species of Anopheles. Previous research has described mosquito behavioural alterations resulting from widespread ITN coverage, which could result in a decrease in net efficacy. Here, behaviours were compared including timings of net contact, willingness to refeed and longevity post-exposure to two next-generation nets, PermaNet ® 3.0 (P3 net) and Interceptor ® G2 (IG2 net) in comparison with a standard pyrethroid-only net (Olyset Net™ (OL net)) and an untreated net., Methods: Susceptible and resistant Anopheles gambiae mosquitoes were exposed to the nets with a human volunteer host in a room-scale assay. Mosquito movements were tracked for 2 h using an infrared video system, collecting flight trajectory, spatial position and net contact data. Post-assay, mosquitoes were monitored for a range of sublethal insecticide effects., Results: Mosquito net contact was focused predominantly on the roof for all four bed nets. A steep decay in activity was observed for both susceptible strains when P3 net and OL net were present and with IG2 net for one of the two susceptible strains. Total mosquito activity was higher around untreated nets than ITNs. There was no difference in total activity, the number, or duration, of net contact, between any mosquito strain, with similar behaviours recorded in susceptible and resistant strains at all ITNs. OL net, P3 net and IG2 net all killed over 90% of susceptible mosquitoes 24 h after exposure, but this effect was not seen with resistant mosquitoes where mortality ranged from 16 to 72%. All treated nets reduced the willingness of resistant strains to re-feed when offered blood 1-h post-exposure, with a more pronounced effect seen with P3 net and OL net than IG2 net., Conclusion: These are the first results to provide an in-depth description of the behaviour of susceptible and resistant Anopheles gambiae strains around next-generation bed nets using a room-scale tracking system to capture multiple behaviours. These results indicate that there is no major difference in behavioural responses between mosquito strains of differing pyrethroid susceptibility when exposed to these new ITNs under the experimental conditions used., (© 2023. The Author(s).)

Background: Since 2000, Burkina Faso has experienced regular dengue cases and outbreaks, making dengue an increasingly important health concern for the country. Previous studies in Burkina Faso reported that resistance of Aedes aegypti to pyrethroid insecticides was associated with the F1534C and V1016I kdr mutations. The current study reports high resistance of Ae. aegypti populations to pyrethroid insecticides, likely supported by mutations in the voltage-gated sodium channel, here evidenced by genotyping the kdr SNPs V410L, V1016I and F1534C. We also describe a new multiplex PCR-based diagnostic of F1534C and V1016I kdr SNPs., Methods: Larvae of Ae. aegypti were collected from three health districts of Ouagadougou in 2018. The resistance status of Ae. aegypti to permethrin (15 μg/ml) and deltamethrin (10 μg/ml) was tested using bottles and to malathion (5%) using WHO tube tests. All bioassays used 1-h exposure and mortality recorded 24 h post-exposure. Bioassay results were interpreted according to WHO thresholds for resistance diagnosis. The kdr mutations were screened using AS-PCR and TaqMan methods in exposed and non-exposed Aedes mosquitoes., Results: Females from all health districts were resistant to permethrin and deltamethrin (< 20% mortality) but were fully susceptible to 5% malathion. The F1534C and V1016I kdr mutations were successfully detected using a newly developed multiplex PCR in perfect agreement with TaqMan method. The 1534C/1016I/410L haplotype was correlated with permethrin resistance but not with deltamethrin resistance; however, the test power was limited by a low frequency of dead individuals in deltamethrin exposure., Conclusions: Resistance to pyrethroid insecticides is associated with kdr mutant haplotypes, while the absence of substantial resistance to malathion suggests that it remains a viable option for dengue vector control in Ouagadougou., (© 2023. The Author(s).)

Introduction: Despite a high fatality rate in humans, little is known about the occurrence of Crimean-Congo hemorrhagic fever virus (CCHFV) in Cameroon. Hence, this pioneer study was started with the aim of determining the prevalence of CCHFV in domestic ruminants and its potential vector ticks in Cameroon., Methods: A cross-sectional study was carried out in two livestock markets of Yaoundé to collect blood and ticks from cattle, sheep, and goats. CCHFV-specific antibodies were detected in the plasma using a commercial ELISA assay and confirmed using a modified seroneutralization test. Ticks were screened for the presence of orthonairoviruses by amplification of a fragment of the L segment using RT-PCR. Phylogeny was used to infer the genetic evolution of the virus., Results: Overall, 756 plasma samples were collected from 441 cattle, 168 goats, and 147 sheep. The seroprevalence of CCHFV was 61.77% for all animals, with the highest rate found in cattle (433/441, 98.18%) followed by sheep (23/147, 15.65%), and goats (11/168, 6.55%), ( p -value < 0.0001). The highest seroprevalence rate was found in cattle from the Far North region (100%). Overall, 1500 ticks of the Rhipicephalus (773/1500, 51.53%), Amblyomma (341/1500, 22.73%), and Hyalomma (386/1500, 25.73%) genera were screened. CCHFV was identified in one Hyalomma truncatum pool collected from cattle. Phylogenetic analysis of the L segment classified this CCHFV strain within the African genotype III., Conclusion: These seroprevalence results call for additional epidemiological studies on CCHFV, especially among at-risk human and animal populations in high-risk areas of the country., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Simo Tchetgna, Yousseu, Cosset, de Freitas, Kamgang, McCall, Ndip, Legros and Wondji.)

Mosquito-borne diseases account for around one million deaths annually. There is a constant need for novel intervention mechanisms to mitigate transmission, especially as current insecticidal methods become less effective with the rise of insecticide resistance among mosquito populations. Previously, we used a near infra-red tracking system to describe the behaviour of mosquitoes at a human-occupied bed net, work that eventually led to an entirely novel bed net design. Advancing that approach, here we report on the use of trajectory analysis of a mosquito flight, using machine learning methods. This largely unexplored application has significant potential for providing useful insights into the behaviour of mosquitoes and other insects. In this work, a novel methodology applies anomaly detection to distinguish male mosquito tracks from females and couples. The proposed pipeline uses new feature engineering techniques and splits each track into segments such that detailed flight behaviour differences influence the classifier rather than the experimental constraints such as the field of view of the tracking system. Each segment is individually classified and the outcomes are combined to classify whole tracks. By interpreting the model using SHAP values, the features of flight that contribute to the differences between sexes are found and are explained by expert opinion. This methodology was tested using 3D tracks generated from mosquito mating swarms in the field and obtained a balanced accuracy of 64.5% and an ROC AUC score of 68.4%. Such a system can be used in a wide variety of trajectory domains to detect and analyse the behaviours of different classes, e.g., sex, strain, and species. The results of this study can support genetic mosquito control interventions for which mating represents a key event for their success.

Aedes aegypti is a ubiquitous vector of arboviruses mostly in urbanised areas throughout the tropics and subtropics and a growing threat beyond. Control of Ae. aegypti is difficult and costly, and no vaccines are available for most of the viruses it transmits. With practical control solutions our goal, ideally suitable for delivery by householders in affected communities, we reviewed the literature on adult Ae. aegypti biology and behaviour, within and close to the human home, the arena where such interventions must impact. We found that knowledge was vague or important details were missing for multiple events or activities in the mosquito life cycle, such as the duration or location of the many periods when females rest between blood feeding and oviposition. The existing body of literature, though substantial, is not wholly reliable, and evidence for commonly held "facts" range from untraceable to extensive. Source references of some basic information are poor or date back more than 60 years, while other information that today is accepted widely as "fact" is not supported by evidence in the literature. Many topics, e.g., sugar feeding, resting preferences (location and duration), and blood feeding, merit being revisited in new geographical regions and ecological contexts to identify vulnerabilities for exploitation in control.

Background: Outbreaks of dengue fever caused by viruses transmitted by Aedes aegypti mosquitoes are repeated occurrences in West Africa. In recent years, Burkina Faso has experienced major dengue outbreaks, most notably in 2016 and 2017 when 80% of cases were recorded in Ouagadougou City (Central Health Region). In order to better understand the ecology of this vector and to provide information for use in developing control measures, a study on the characteristics of Aedes container breeding sites and the productivity of such sites, as measured by the abundance of immature stages and resultant adult body size, was undertaken in three health districts (Baskuy, Bogodogo and Nongremassom) of Ouagadougou., Methods: Adult mosquitoes were collected indoors and outdoors in 643 households during the rainy season from August to October 2018. The presence of water containers was systematically recorded and the containers examined for the presence or absence of larvae. Characteristics of the container breeding sites, including size of the container and temperature, pH and conductivity of the water contained within, were recorded as well as the volume of water. Traditional Stegomyia indices were calculated as quantitative indicators of the risk of dengue outbreaks; generalised mixed models were fitted to larval and pupal densities, and the contribution of each covariate to the model was evaluated by the Z-value and associated P-value., Results: A total of 1061 container breeding sites were inspected, of which 760 contained immature stages of Ae. aegypti ('positive' containers). The most frequent container breeding sites found in each health district were tyres and both medium (buckets/cans/pots) and large (bins/barrels/drums) containers; these containers were also the most productive larval habitats and the types that most frequently tested positive. Of the Stegomyia indices, the Breteau, House and Container indices exceeded WHO dengue risk thresholds. Generalised linear mixed models showed that larval and pupal abundances were associated with container type, physicochemical characteristics of the water and collection month, but there were significant differences among container types and among health districts. Aedes aegypti body size was positively associated with type and diameter of the container, as well as with electrical conductivity of the water, and negatively associated with pH and temperature of the water and with the level of exposure of the container to sunlight., Conclusion: This study provides data on putative determinants of the productivity of habitats regarding Ae. aegypti immature stages. These data are useful to better understand Ae. aegypti proliferation. The results suggest that identifying and targeting the most productive container breeding sites could contribute to dengue vector control strategies in Burkina Faso., (© 2022. The Author(s).)

Little is known about the impact of ticks on livestock and humans in Cameroon. This study aimed to determine the prevalence, seasonal variation, and genetic diversity of hard ticks in the country. Ticks were collected during a cross-sectional survey on domestic livestock in two markets of Yaoundé in 2019 and 2020 and identified using morphological keys, 16S ribosomal DNA, (16S rDNA), and the cytochrome c oxidase subunit 1 (Cox1) genes. The infestation rates were 39.18%, 11.53%, and 2.74% in cattle, sheep, and goats respectively. Three genera of ticks were identified, Rhipicephalus, Amblyomma, and Hyalomma comprising eleven tick species. The main species were Rhipicephalus decoloratus (30.25%), R. microplus (24.43%), and Amblyomma variegatum (12.96%). Rhipicephalus spp. (81.31%) and Amblyomma variegatum (51.54%) were abundant during the rainy season, while Hyalomma spp. (83.86%) during the dry season (p-value <0.00001). Cox1 and 16S rDNA analysis showed a high level of genetic diversity among tick species with sequences close to those observed across Africa. Phylogenetic analysis revealed that our R. microplus belong to clade A and we identified R. sanguineus s.l. as R. linnea. This study shows a high tick infestation rate in cattle, while low in small ruminants with an extensive diversity of tick species, including several known vectors of important tick-borne diseases., (© 2022 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

The response to recent dengue outbreaks in Burkina Faso was insecticide-based, despite poor knowledge of the vector population's susceptibility to the insecticides used. Here, we report on the susceptibility to the main insecticide classes and identify important underlying mechanisms in Aedes aegypti populations in Ouagadougou and Banfora, in 2019 and 2020. Wild Ae. aegypti were tested as adults in WHO bioassays and then screened in real time melting curve qPCR analyses to genotype the F1534C, V1016I, and V410L Aedes kdr mutations. Ae. aegypti showed moderate resistance to 0.1% bendiocarb (80-95% survival post-exposure), 0.8% Malathion (60-100%), 0.21% pirimiphos-methyl (75% - 97%), and high resistance to 0.03% deltamethrin (20-70%). PBO pre-exposure partially restored pyrethroid susceptibility. Genotyping detected high frequency of 1534C allele (0.92) and moderate 1016I (0.1-0.32). The V410L mutation was detected in Burkina Faso for the first time (frequency 0.1-0.36). Mosquitoes surviving 4 h exposure to 0.03% deltamethrin had significantly higher frequencies of the F1534C mutation than dead mosquitoes (0.70 vs. 0.96, p < 0.0001) and mosquitoes surviving 2 - 4 h exposure had a significantly reduced life span. Ae. aegypti from Burkina Faso are resistant to multiple insecticide classes with multiple mechanisms involved, demonstrating the essential role of insecticide resistance monitoring within national dengue control programmes., (© 2022 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

Despite the ethical issues concerning the continued use of Human Landing Catches (HLC) to monitor the Simulium damnosum complex for epidemiological monitoring of onchocericasis, few attempts to develop alternatives have been reported. In studies on a wild population of S. damnosum in Burkina Faso, we tested visual targets (different sizes and shapes) and olfactory stimuli (CO 2 , and POCA and BG-lure® odour blends) for their ability to attract and collect host seeking blackflies. At each trap, blackflies were caught with appropriately sized electrocuting grids and results from Latin square design tests were compared. Throughout, HLCs captured more blackflies than the targets. Of the traps tested, small targets (0.0625 and 0.5 m 2 ) were the most efficient visual lure in terms of the number of S. damnosum captured per unit area 1.7-5× more than larger targets. Overall, results suggested that sticky black targets of horizontal rectangular shape (0.125-0.5 m 2 ) and baited with a POCA and/or CO 2 mixture could provide a cheap practical field alternative to HLC for onchocerciasis xenomonitoring, subject to confirmation that the design has no inherent bias for certain members of the S. damnosum species complex., (© 2022 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

Background: Rift Valley fever (RVF) is a mosquito-borne zoonosis endemic in Africa. With little known of the burden or epidemiology of RVF virus (RVFV) in Cameroon, this study aimed to determine the seroprevalence of RVFV in domestic ruminants of various origins in two markets of Yaoundé, Cameroon., Methodology/principal Findings: The origin of animals randomly sampled at two livestock markets in Yaoundé were recorded and plasma samples collected for competitive and capture Enzyme-linked Immunosorbent Assay (ELISA) to determine the prevalence of Immunoglobulins G (IgG) and Immunoglobulins M (IgM) antibodies. Following ELISA IgM results, a real-time reverse transcription-polymerase chain reaction (qRT-PCR) was performed to detect RVFV RNA. In June-August 2019, February-March 2020, and March-April 2021, 756 plasma samples were collected from 441 cattle, 168 goats, and 147 sheep. RVFV IgG seroprevalence was 25.7% for all animals, 42.2% in cattle, 2.7% in sheep, and 2.4% in goats. However, IgM seroprevalence was low, at 0.9% in all animals, 1.1% in cattle, 1.4% in sheep, and 0% in goats. The seroprevalence rates varied according to the animal's origin with the highest rate (52.6%) in cattle from Sudan. In Cameroon, IgG and IgM rates respectively were 45.1% and 2.8% in the North, 44.8% and 0% in the Adamawa, 38.6% and 1.7% in the Far-North. All IgM positive samples were from Cameroon. In cattle, 2/5 IgM positive samples were also IgG positive, but both IgM positive samples in sheep were IgG negative. Three (42.9%) IgM positive samples were positive for viral RVFV RNA using qRT-PCR but given the high ct values, no amplicon was obtained., Conclusion/significance: These findings confirm the circulation of RVFV in livestock in Cameroon with prevalence rates varying by location. Despite low IgM seroprevalence rates, RVF outbreaks can occur without being noticed. Further epidemiological studies are needed to have a broad understanding of RVFV transmission in Cameroon., Competing Interests: The authors have declared that no competing interests exist.

Background: Dengue's emergence in West Africa was typified by the Burkina Faso outbreaks in 2016 and 2017, the nation's largest to date. In both years, we undertook three-month surveys of Aedes populations in or near the capital city Ouagadougou, where the outbreaks were centered., Methodology: In 1200LG (urban), Tabtenga (peri-urban) and Goundry (rural) localities, we collected indoor and outdoor resting mosquito adults, characterized larval habitats and containers producing pupae and reared immature stages to adulthood in the laboratory for identification. All mosquito adults were identified morphologically. Host species (from which bloodmeals were taken) were identified by PCR. Generalized mixed models were used to investigate relationships between adult or larval densities and multiple explanatory variables., Results: From samples in 1,780 houses, adult Ae. aegypti were significantly more abundant in the two urban localities (Tabtenga and 1200 LG) in both years than in the rural site (Goundry), where Anopheles spp. were far more common. Results from adult collections indicated a highly exophilic and anthropophilic (>90% bloodmeals of human origin) vector population, but with a relatively high proportion of bloodfed females caught inside houses. Habitats producing most pupae were waste tires (37% of total pupae), animal troughs (44%) and large water barrels (30%). While Stegomyia indices were not reliable indicators of adult mosquito abundance, shared influences on adult and immature stage densities included rainfall and container water level, collection month and container type/purpose. Spatial analysis showed autocorrelation of densities, with a partial overlap in adult and immature stage hotspots., Conclusion: Results provide an evidence base for the selection of appropriate vector control methods to minimize the risk, frequency and magnitude of future outbreaks in Ouagadougou. An integrated strategy combining community-driven practices, waste disposal and insecticide-based interventions is proposed. The prospects for developing a regional approach to arbovirus control in West Africa or across Africa are discussed., Competing Interests: The authors have declared that no competing interests exist.

Cardiovascular complications due to COVID-19, such as right ventricular dysfunction, are common. The combination of acute respiratory distress syndrome, invasive mechanical ventilation, thromboembolic disease and direct myocardial injury creates conditions where right ventricular dysfunction is likely to occur. We undertook a prospective, multicentre cohort study in 10 Scottish intensive care units of patients with COVID-19 pneumonitis whose lungs were mechanically ventilated. Right ventricular dysfunction was defined as the presence of severe right ventricular dilation and interventricular septal flattening. To explore the role of myocardial injury, high-sensitivity troponin and N-terminal pro B-type natriuretic peptide plasma levels were measured in all patients. We recruited 121 patients and 118 (98%) underwent imaging. It was possible to determine the primary outcome in 112 (91%). Severe right ventricular dilation was present in 31 (28%), with interventricular septal flattening present in nine (8%). Right ventricular dysfunction (the combination of these two parameters) was present in seven (6%, 95%CI 3-13%). Thirty-day mortality was 86% in those with right ventricular dysfunction as compared with 45% in those without (p = 0.051). Patients with right ventricular dysfunction were more likely to have: pulmonary thromboembolism (p < 0.001); higher plateau airway pressure (p = 0.048); lower dynamic compliance (p = 0.031); higher plasma N-terminal pro B-type natriuretic peptide levels (p = 0.006); and raised plasma troponin levels (p = 0.048). Our results demonstrate a prevalence of right ventricular dysfunction of 6%, which was associated with increased mortality (86%). Associations were also observed between right ventricular dysfunction and aetiological domains of: acute respiratory distress syndrome; ventilation; thromboembolic disease; and direct myocardial injury, implying a complex multifactorial pathophysiology., (© 2022 The Authors. Anaesthesia published by John Wiley & Sons Ltd on behalf of Association of Anaesthetists.)

Background: The WHO cone test is one of three tests currently used to evaluate the efficacy of insecticide-treated bed nets (ITNs). It generates two test outputs, knockdown and 24-h mortality, both indicative of immediate toxicity but that reveal little about the nature of mosquito and ITN interaction or how results translate to real-world settings., Methods: A human arm held 5 mm behind the net surface acted as a host attractant during cone tests and a smartphone was used to capture mosquito behaviour in the cone. Post-exposure blood feeding and survival for nine days were recorded; ingested blood meal size was determined by measuring excreted haematin. Four strains of Anopheles gambiae (insecticide susceptible: Kisumu and N'gousso; insecticide resistant: Banfora and VK7) were tested with and without the host attractant using untreated, Permanet 2.0 and Olyset nets. Video recordings were scan sampled every five seconds to record mosquito positions on either the net, in flight or in contact with the cone. Generalized estimating equations were used to analyse all data except survival within nine days which was analysed using Weighted Cox Regression., Results: Net contact was the most frequently recorded behaviour in all Anopheles spp. strains on all nets. Adding the human host as attractant triggered excitatory behaviours: in all strains, the magnitude of net contact was significantly decreased compared to tests without a host. ITN exposure altered the observed behaviour of the two susceptible strains, which exhibited a decreased response to the host during ITN tests. The resistant strains did not alter their behaviour during ITN tests. Significantly less net contact was observed during Olyset Net tests compared to Permanet 2.0. The host presence affected survival after exposure: Banfora and VK7 mosquitoes exposed to Permanet 2.0 with a host lived longer compared to tests performed without a host. However, mosquitoes that blood-fed and survived long enough to digest the blood meal did not exhibit significantly reduced longevity regardless of the presence of the host attractant., Conclusions: Simple modifications to the WHO cone test and extension of post-test monitoring beyond the current 24 h enable detailed behavioural characterizations of individual ITNs to be compiled. The effects observed from testing with a host and including blood feeding suggest that more representative estimates of true of ITN efficacy are gained with these modifications than when using the current testing protocol., (© 2022. The Author(s).)

Background: Following earlier trials indicating that their potential in dengue vector control was constrained by housing structure, a large-scale cluster-randomized trial of insecticide treated curtains (ITCs) and water jar covers (ITJCs) was undertaken in Venezuela., Methods: In Trujillo, Venezuela, 60 clusters (6223 houses total) were randomized so that 15 clusters each received either PermaNet insecticide-treated window curtains (ITCs), permanent insecticide-treated water storage jar covers (ITJCs), a combination of both ITCs and ITJCs, or no insecticide treated materials (ITMs). A further 15 clusters located at least 5km from the edge of the study site were selected to act as an external control. Entomological surveys were carried out immediately before and after intervention, and then at 6-month intervals over the following 27 months. The Breteau and House indices were used as primary outcome measures and ovitrap indices as secondary. Negative binomial regression models were used to compare cluster-level values of these indices between the trial arms., Results: Reductions in entomological indices followed deployment of all ITMs and throughout the trial, indices in the external control arm remained substantially higher than in the ITM study arms including the internal control. Comparing the ratios of between-arm means to summarise the entomological indices throughout the study, the combined ITC+ITJC intervention had the greatest impact on the indices, with a 63% difference in the pupae per person indices between the ITC+ITJC arm and the internal control. However, coverage had fallen below 60% by 14-months post-intervention and remained below 40% for most of the remaining study period., Conclusions: ITMs can impact dengue vector populations in the long term, particularly when ITCs and ITJCs are deployed in combination., Trial Registration: ClinicalTrials.gov ISRCTN08474420; www.isrctn.com., Competing Interests: The authors have declared that no competing interests exist.

Western Africa is vulnerable to arboviral disease transmission, having recently experienced major outbreaks of chikungunya, dengue, yellow fever and Zika. However, there have been relatively few studies on the natural history of the two major human arbovirus vectors in this region, Aedes aegypti and Ae. albopictus , potentially limiting the implementation of effective vector control. We systematically searched for and reviewed relevant studies on the behaviour and ecology of Ae. aegypti and Ae. albopictus in Western Africa, published over the last 40 years. We identified 73 relevant studies, over half of which were conducted in Nigeria, Senegal, or Côte d'Ivoire. Most studies investigated the ecology of Ae. aegypti and Ae. albopictus , exploring the impact of seasonality and land cover on mosquito populations and identifying aquatic habitats. This review highlights the adaptation of Ae. albopictus to urban environments and its invasive potential, and the year-round maintenance of Ae. aegypti populations in water storage containers. However, important gaps were identified in the literature on the behaviour of both species, particularly Ae. albopictus . In Western Africa, Ae. aegypti and Ae. albopictus appear to be mainly anthropophilic and to bite predominantly during the day, but further research is needed to confirm this to inform planning of effective vector control strategies. We discuss the public health implications of these findings and comment on the suitability of existing and novel options for control in Western Africa., Competing Interests: Declaration of competing interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acute febrile patients presenting at hospitals in Douala, Cameroon between July and December 2020, were screened for dengue infections using real time RT-PCR on fragments of the 5' and 3' UTR genomic regions. In total, 12.8% (41/320) of cases examined were positive for dengue. Dengue virus 3 (DENV-3) was the most common serotype found (68.3%), followed by DENV-2 (19.5%) and DENV-1 (4.9%). Co-infections of DENV-3 and DENV-2 were found in 3 cases. Jaundice and headache were the most frequent clinical signs associated with infection and 56% (23/41) of the cases were co-infections with malaria. Phylogenetic analysis of the envelope gene identified DENV-1 as belonging to genotype V, DENV-2 to genotype II and DENV-3 to genotype III. The simultaneous occurrence of three serotypes in Douala reveals dengue as a serious public health threat for Cameroon and highlights the need for further epidemiological studies in the major cities of this region., Competing Interests: The authors have declared that no competing interests exist.

Pyrethroid resistance is widespread in malaria vectors. However, differential mortality in discriminating dose assays to different pyrethroids is often observed in wild populations. When this occurs, it is unclear if this differential mortality should be interpreted as an indication of differential levels of susceptibility within the pyrethroid class, and if so, if countries should consider selecting one specific pyrethroid for programmatic use over another. A review of evidence from molecular studies, resistance testing with laboratory colonies and wild populations, and mosquito behavioural assays were conducted to answer these questions. Evidence suggested that in areas where pyrethroid resistance exists, different results in insecticide susceptibility assays with specific pyrethroids currently in common use (deltamethrin, permethrin, α-cypermethrin, and λ-cyhalothrin) are not necessarily indicative of an operationally relevant difference in potential performance. Consequently, it is not advisable to use rotation between these pyrethroids as an insecticide-resistance management strategy. Less commonly used pyrethroids (bifenthrin and etofenprox) may have sufficiently different modes of action, though further work is needed to examine how this may apply to insecticide resistance management.

Background: The efficacy of insecticide-treated nets (ITNs) containing the insect growth regulator pyriproxyfen (PPF) and pyrethroid insecticides (PPF-ITNs) is being assessed in clinical trials to determine whether they provide greater protection from malaria than standard pyrethroid-treated ITNs in areas where mosquitoes are resistant to pyrethroids. Understanding the entomological mode of action of this new ITN class will aide interpretation of the results from these trials., Methods: Anopheles gambiae sensu lato (s.l.) mosquitoes from a susceptible laboratory strain were exposed to PPF-treated netting 24 h, 6 h, and immediately prior to, or 24 h post blood feeding, and the impact on fecundity, fertility and longevity recorded. Pyrethroid-resistant populations were exposed to nets containing permethrin and PPF (PPF-ITNs) in cone bioassays and daily mortality recorded. Mosquitoes were also collected from inside houses pre- and post-distribution of PPF-ITNs in a clinical trial conduced in Burkina Faso; female An. gambiae s.l. were then assessed for fecundity and fertility., Results: PPF exposure reduced the median adult lifespan of insecticide-susceptible mosquitoes by 4 to 5 days in all exposure times (p < 0.05) other than 6 h pre-blood meal and resulted in almost complete lifelong sterilization. The longevity of pyrethroid-resistant mosquitoes was also reduced by at least 5 days after exposure to PPF-ITNs compared to untreated nets, but was unaffected by exposure to standard pyrethroid only ITNs. A total of 386 blood-fed or gravid An. gambiae s.l. females were collected from five villages between 1 and 12 months before distribution of PPF-ITNs. Of these mosquitoes, 75% laid eggs and the remaining 25% appeared to have normal ovaries upon dissection. In contrast, only 8.6% of the 631 blood-fed or gravid An. gambiae s.l. collected post PPF-ITN distribution successfully oviposited; 276 (43.7%) did not oviposit but had apparently normal ovaries upon dissection, and 301 (47.7%) did not oviposit and had abnormal eggs upon dissection. Egg numbers were also significantly lower (average of 138/female prior distribution vs 85 post distribution, p < 0.05)., Conclusion: Exposure to a mixture of PPF and pyrethroids on netting shortens the lifespan of mosquitoes and reduces reproductive output. Sterilization of vectors lasted at least one year under operational conditions. These findings suggest a longer effective lifespan of PPF-pyrethroid nets than reported previously.

Microbiome research has gained considerable interest due to the emerging evidence of its impact on human and animal health. As in other animals, the gut-associated microbiota of mosquitoes affect host fitness and other phenotypes. It is now well established that microbes can alter pathogen transmission in mosquitoes, either positively or negatively, and avenues are being explored to exploit microbes for vector control. However, less attention has been paid to how microbiota affect phenotypes that impact vectorial capacity. Several mosquito and pathogen components, such as vector density, biting rate, survival, vector competence, and the pathogen extrinsic incubation period all influence pathogen transmission. Recent studies also indicate that mosquito gut-associated microbes can impact each of these components, and therefore ultimately modulate vectorial capacity. Promisingly, this expands the options available to exploit microbes for vector control by also targeting parameters that affect vectorial capacity. However, there are still many knowledge gaps regarding  mosquito-microbe interactions  that need to be addressed in order to exploit them efficiently. Here, we review current evidence of impacts of the microbiome on aspects of vectorial capacity, and we highlight likely opportunities for novel vector control strategies and areas where further studies are required. Video abstract.

Background: Advances in digitized video-tracking and behavioural analysis have enabled accurate recording and quantification of mosquito flight and host-seeking behaviours, facilitating development of individual (agent) based models at much finer spatial scales than previously possible., Methods: Quantified behavioural parameters were used to create a novel virtual testing model, capable of accurately simulating indoor flight behaviour by a virtual population of host-seeking mosquitoes as they interact with and respond to simulated stimuli from a human-occupied bed net. The model is described, including base mosquito behaviour, state transitions, environmental representation and host stimulus representation., Results: In the absence of a bed net and human host bait, flight distribution of the model population was relatively uniform throughout the arena. Introducing an unbaited untreated bed net induced a change in distribution with an increase in landing events on the net surface, predominantly on the sides of the net. Adding the presence of a simulated human bait dramatically impacted flight distribution patterns, exploratory foraging and, the number and distribution of landing positions on the net, which were determined largely by the orientation of the human within. The model replicates experimental results with free-flying living mosquitoes at human-occupied bed nets, where contact occurs predominantly on the top surface of the net. This accuracy is important as it quantifies exposure to the lethal insecticide residues that may be unique to the net roof (or theoretically any other surface). Number of net contacts and height of contacts decreased with increasing attractant dispersal noise., Conclusions: Results generated by the model are an accurate representation of actual mosquito behaviour recorded at and around a human-occupied bed net in untreated and insecticide-treated nets. This fine-grained model is highly flexible and has significant potential for in silico screening of novel bed net designs, potentially reducing time and cost and accelerating the deployment of new and more effective tools for protecting against malaria in sub-Saharan Africa.

Vector control methods that mobilize and impact rapidly during dengue, Zika, and chikungunya outbreaks are urgently needed in urban contexts. We investigated whether one person using a handheld aerosolized insecticide could achieve efficacy levels comparable to targeted indoor residual spraying (TIRS), using pyrethroid-resistant Aedes aegypti in a semi-field setting with experimental houses in Mexico. The insecticide product (H24, a carbamate and pyrethroid mixture), available over-the-counter locally, was sprayed only on known Ae. aegypti -resting surfaces, for example, walls less than 1.5 m and dark hidden areas. In six identical houses with paired bedrooms, one bedroom was treated, and the other remained an untreated control. Each week for 8 weeks, 100 female pyrethroid-resistant Ae. aegypti were released in each bedroom and followed up daily. Mortality rates in treated bedrooms exceeded 90% for at least 2 weeks, and more than 80% (89.2; 95% CI: 79.98-98.35) for 3 weeks or more. Mortality rates in control houses were zero. Results demonstrate that the immediate impact of TIRS can be delivered by one person using existing products, at an estimated cost for the average household in Mexico of under US$3 per month. Triggered by early outbreak signs, dissemination via community hubs and mass/social media of instructions to treat the home immediately, with monthly re-treatment thereafter, provides a simple means to engage and empower householders. Compatible with integrated vector management strategies, it enables self-protection even if existing agencies falter, a situation exemplified by the potential impact on vector control of the restrictions imposed during the 2020 COVID-19 pandemic.

Mosquito-borne Zika virus (ZIKV) transmission has almost exclusively been detected in the tropics despite the distributions of its primary vectors extending farther into temperate regions. Therefore, it is unknown whether ZIKV's range has reached a temperature-dependent limit, or if it can spread into temperate climates. Using field-collected mosquitoes for biological relevance, we found that two common temperate mosquito species, Aedes albopictus and Ochlerotatus detritus , were competent for ZIKV. We orally exposed mosquitoes to ZIKV and held them at between 17 and 31°C, estimated the time required for mosquitoes to become infectious, and applied these data to a ZIKV spatial risk model. We identified a minimum temperature threshold for the transmission of ZIKV by mosquitoes between 17 and 19°C. Using these data, we generated standardized basic reproduction number R 0 -based risk maps and we derived estimates for the length of the transmission season for recent and future climate conditions. Our standardized R 0 -based risk maps show potential risk of ZIKV transmission beyond the current observed range in southern USA, southern China and southern European countries. Transmission risk is simulated to increase over southern and Eastern Europe, northern USA and temperate regions of Asia (northern China, southern Japan) in future climate scenarios.

Robust imaging techniques for tracking insects have been essential tools in numerous laboratory and field studies on pests, beneficial insects and model systems. Recent innovations in optical imaging systems and associated signal processing have enabled detailed characterization of nocturnal mosquito behaviour around bednets and improvements in bednet design, a global essential for protecting populations against malaria. Nonetheless, there remain challenges around ease of use for large-scale in situ recordings and extracting data reliably in the critical areas of the bednet where the optical signal is attenuated. Here, we introduce a retro-reflective screen at the back of the measurement volume, which can simultaneously provide diffuse illumination, and remove optical alignment issues while requiring only one-sided access to the measurement space. The illumination becomes significantly more uniform, although noise removal algorithms are needed to reduce the effects of shot noise, particularly across low-intensity bednet regions. By systematically introducing mosquitoes in front of and behind the bednet in laboratory experiments, we are able to demonstrate robust tracking in these challenging areas. Overall, the retro-reflective imaging set-up delivers mosquito segmentation rates in excess of 90% compared to less than 70% with backlit systems., Competing Interests: The authors declare no competing interests., (© 2020 The Authors.)

Dengue is one of the most important vector-borne diseases, resulting in an estimated hundreds of millions of infections annually throughout the tropics. Control of dengue is heavily dependent upon control of its primary mosquito vector, Aedes aegypti. Innovative interventions that are effective at targeting the adult stage of the mosquito are needed to increase the options for effective control. The use of insecticide-treated curtains (ITCs) has previously been shown to significantly reduce the abundance of Ae. aegypti in and around homes, but the impact of ITCs on dengue virus (DENV) transmission has not been rigorously quantified. A parallel arm cluster-randomized controlled trial was conducted in Iquitos, Peru to quantify the impact of ITCs on DENV seroconversion as measured through plaque-reduction neutralization tests. Seroconversion data showed that individuals living in the clusters that received ITCs were at greater risk to seroconverting to DENV, with an average seroconversion rate of 50.6 per 100 person-years (PY) (CI: 29.9-71.9), while those in the control arm had an average seroconversion rate of 37.4 per 100 PY (CI: 15.2-51.7). ITCs lost their insecticidal efficacy within 6 months of deployment, necessitating re-treatment with insecticide. Entomological indicators did not show statistically significant differences between ITC and non-ITC clusters. It's unclear how the lack of protective efficacy reported here is attributable to simple failure of the intervention to protect against Ae. aegypti bites, or the presence of a faulty intervention during much of the follow-up period. The higher risk of dengue seroconversion that was detected in the ITC clusters may have arisen due to a false sense of security that inadvertently led to less routine protective behaviors on the part of households that received the ITCs. Our study provides important lessons learned for conducting cluster randomized trials for vector control interventions against Aedes-transmitted virus infections., Competing Interests: The authors have declared that no competing interests exist.

Background: Insecticide-treated nets (ITNs) are losing efficacy against pyrethroid-resistant malaria vector populations throughout Africa. Safeguarding bed net efficacy, vital for effective malaria control, requires greater knowledge of mosquito-ITN interactions and how this impacts on the mosquito., Methods: A purpose-built benchtop apparatus with a closed 10 cm cubic chamber (the 'Baited-box') was used to video record behaviour of individual free-flying female Anopheles gambiae during approach and blood-feeding on a human hand through untreated nets and ITNs at close range. Time and duration of defined behavioural events, and knockdown and mortality at 1- and 24-h post-exposure respectively, were recorded for pyrethroid susceptible and resistant mosquitoes., Results: Using three human volunteers differing in relative attractiveness to mosquitoes, 328 mosquitoes were individually tested. There were no significant differences between response rates to ITNs and untreated nets (P > 0.1) or between resistant (Tiassalé) and susceptible (Kisumu) mosquito strains, at untreated nets (P = 0.39) or PermaNet 2.0 (P = 1). The sequence of behavioural events from host-seeking to completion of blood-feeding was consistent in all tests but duration and start time of events involving net contact were reduced or delayed respectively with ITNs. Blood-feeding durations at untreated nets (means from 4.25 to 8.47 min (95% confidence interval (CI) = 3.39-9.89) at 3 human volunteers) were reduced by 37-50% at PermaNet 2.0, in susceptible (mean 2.59-4.72 min, 95% CI = 1.54-5.5, P = < 0.05) and resistant (mean 4.20 min, 95% CI = 3.42-4.97, P = 0.01) strains. Total accumulated net contact was approximately 50% lower at PermaNet and Olyset ITNs (P < 0.0001) in susceptible (two of the three volunteers) and resistant mosquitoes. Times prior to first net contact were similar at untreated nets and ITNs (P > 0.2), and neither ITN type showed detectable spatial repellency. After initial contact, blood-feeding commenced later at Olyset (mean 2.76 min, 95% CI = 1.74-3.76, P = 0.0009) and PermaNet (mean 2.4 min, 95% CI = 1.52-3.33, P = 0.0058) than untreated netting (mean 0.68 min, 95% CI = 0.42-0.94)., Conclusions: The baited box offers a simple method for detailed characterization of mosquito behavioural responses to insecticidal nets, for comparing entomological modes of action between nets and for defining the behavioural responses of particular mosquito strains or populations. The device has potential as a screening assay in the search for novel net treatments and for investigations into behavioural resistance mechanisms.

Transmission of Plasmodium falciparum malaria parasites occurs when nocturnal Anopheles mosquito vectors feed on human blood. In Africa, where malaria burden is highest, bednets treated with pyrethroid insecticide were highly effective in preventing mosquito bites and reducing transmission, and essential to achieving unprecedented reductions in malaria until 2015 (ref. 1 ). Since then, progress has stalled 2 , and with insecticidal bednets losing efficacy against pyrethroid-resistant Anopheles vectors 3,4 , methods that restore performance are urgently needed to eliminate any risk of malaria returning to the levels seen before their widespread use throughout sub-Saharan Africa 5 . Here, we show that the primary malaria vector Anopheles gambiae is targeted and killed by small insecticidal net barriers positioned above a standard bednet in a spatial region of high mosquito activity but zero contact with sleepers, opening the way for deploying many more insecticides on bednets than is currently possible. Tested against wild pyrethroid-resistant A. gambiae in Burkina Faso, pyrethroid bednets with organophosphate barriers achieved significantly higher killing rates than bednets alone. Treated barriers on untreated bednets were equally effective, without significant loss of personal protection. Mathematical modelling of transmission dynamics predicted reductions in clinical malaria incidence with barrier bednets that matched those of 'next-generation' nets recommended by the World Health Organization against resistant vectors. Mathematical models of mosquito-barrier interactions identified alternative barrier designs to increase performance. Barrier bednets that overcome insecticide resistance are feasible using existing insecticides and production technology, and early implementation of affordable vector control tools is a realistic prospect.

Unplanned intensive care admission is a devastating complication of lung resection and is associated with significantly increased mortality. We carried out a two-year retrospective national multicentre cohort study to investigate the influence of anaesthetic and analgesic technique on the need for unplanned postoperative intensive care admission. All patients undergoing lung resection surgery in 16 thoracic surgical centres in the UK in the calendar years 2013 and 2014 were included. We defined critical care admission as the unplanned need for either tracheal intubation and mechanical ventilation or renal replacement therapy, and sought an association between mode of anaesthesia (total intravenous anaesthesia vs. volatile) and analgesic technique (epidural vs. paravertebral) and need for intensive care admission. A total of 253 out of 11,208 patients undergoing lung resection in the study period had an unplanned admission to intensive care in the postoperative period, giving an incidence of intensive care unit admission of 2.3% (95%CI 2.0-2.6%). Patients who had an unplanned admission to intensive care unit had a higher mortality (29.00% vs. 0.03%, p < 0.001), and hospital length of stay was increased (26 vs. 6 days, p < 0.001). Across univariate, complete case and multiple imputation (multivariate) models, there was a strong and significant effect of both anaesthetic and analgesic technique on the need for intensive care admission. Patients receiving total intravenous anaesthesia (OR 0.50 (95%CI 0.34-0.70)), and patients receiving epidural analgesia (OR 0.56 (95%CI 0.41-0.78)) were less likely to have an unplanned admission to intensive care after thoracic surgery. This large retrospective study suggests a significant effect of both anaesthetic and analgesic technique on outcome in patients undergoing lung resection. We must emphasise that the observed association does not directly imply causation, and suggest that well-conducted, large-scale randomised controlled trials are required to address these fundamental questions., (© 2019 Association of Anaesthetists.)

Objectives: Lung cancer is a leading cause of cancer death and in suitable cases the best chance of cure is offered by surgery. Lung resection is associated with significant postoperative cardiorespiratory morbidity, with dyspnea and reduced functional capacity as dominant features. These changes are poorly associated with deterioration in pulmonary function and a potential role of right ventricular (RV) dysfunction has been hypothesized. Cardiovascular magnetic resonance imaging is a reference method for noninvasive assessment of RV function and has not previously been applied to this population., Methods: We used cardiovascular magnetic resonance imaging to assess the RV response to lung resection. Cardiovascular magnetic resonance imaging with volume and flow analysis was performed on 27 patients preoperatively, on postoperative day 2 and at 2 months. Left ventricular ejection fraction and RV ejection fraction, the ratio of stroke volume to end systolic volume, pulmonary artery acceleration time, and distensibility of main and branch pulmonary arteries were studied., Results: Mean ± standard deviation RV ejection fraction deteriorated from 50.5% ± 6.9% preoperatively to 45.6% ± 4.5% on postoperative day 2 and remained depressed at 44.9% ± 7.7% by 2 months (P = .003). The ratio of stroke volume to end systolic volume deteriorated from median 1.0 (quartile 1, quartile 3: 0.9, 1.2) preoperatively to median 0.8 (quartile 1, quartile 3: 0.7, 1.0) on postoperative day 2 (P = .011). On postoperative day 2 there was a decrease in pulmonary artery acceleration time and operative pulmonary artery distensibility (P < .030 for both). There were no changes in left ventricular ejection fraction during the study period (P = .621)., Conclusions: These findings suggest RV dysfunction occurs following lung resection and persists 2 months after surgery. The deterioration in the ratio of stroke volume to end systolic volume suggests a mismatch between afterload and contractility. There is an increase in indices of pulsatile afterload resulting from the operative pulmonary artery., (Copyright © 2019 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.)

Context: Right ventricular (RV) dysfunction occurs after lung resection and is associated with postoperative morbidity. Noninvasive evaluation of the RV is challenging, particularly in the postoperative period. A reliable measure of RV function would have value in this population., Aims: This study compares eccentricity index (EI) obtained by transthoracic echocardiography (TTE) with cardiovascular magnetic resonance (CMR) determined measures of RV function in a lung resection cohort. CMR is the reference method for noninvasive assessment of RV function., Design and Setting: Prospective observational cohort study at a single tertiary hospital., Materials and Methods: Twenty-eight patients scheduled for elective lung resection underwent contemporaneous TTE and CMR imaging preoperatively, on postoperative day (POD) 2 and at 2-month. Systolic and diastolic EI was measured offline from anonymized and randomized TTE and CMR images., Statistical Analysis: Bland-Altman analysis was performed to determine agreement between EI TTE and EI CMR . Changes over time and comparison with CMR determined RV ejection fraction (RVEF CMR ) was assessed., Results: Bland-Altman analysis showed a negligible mean difference between EI TTE and EI CMR , but limits of agreement were wide (SD 0.24 and 0.28). There were no significant changes in EI TTE and EI CMR over time ( P > 0.35). We found no association between EI TTE with RVEF CMR at all-time points ( P > 0.22). Systolic and diastolic EI CMR on POD 2 demonstrated moderate association with RVEF CMR ( r = -0.54 and r = -0.59, P ≤ 0.01). At 2-month, only diastolic EI CMR correlated with RVEF CMR ( r = -0.43, P = 0.03). There were no meaningful associations between EI TTE and EI CMR with TTE-derived RV systolic pressure ( P > 0.31)., Conclusions: TTE determined EI is not useful as a noninvasive method of assessing RV function following lung resection., Competing Interests: There are no conflicts of interest., (Copyright: © 2019 Journal of Cardiovascular Echography.)

Objectives: Following lung resection, there is a decrease in the functional capacity and quality of life, which is not fully explained by changes in pulmonary function. Previous work demonstrates that B-type natriuretic peptide (BNP) is associated with short- and long-term complications following lung resection, leading to the suggestion that cardiac dysfunction may contribute to functional deterioration. Our aim was to investigate any relationship between BNP and subjective and objective indices of functional deterioration following lung resection surgery., Methods: Twenty-seven patients undergoing lung resection had serum BNP measured preoperatively, on postoperative day (POD)1 and POD2, and at 2 months postoperatively. The functional deterioration was assessed using 6-min walk tests and the Medical Research Council dyspnoea scale. 'Deterioration in functional capacity' was defined as either an increase in the Medical Research Council dyspnoea score or a significant decrease in the 6-min walk test distance., Results: BNP increased over time (P < 0.01) and was significantly elevated on POD1 and POD2 (P < 0.02 for both). Seventeen patients demonstrated functional deterioration 2 months postoperatively. At all perioperative time points, BNP was significantly higher in patients showing deterioration (P < 0.05 for all). Preoperative BNP was predictive of functional deterioration at 2 months with an area under the receiver-operating characteristic curve of 0.82 (P = 0.01, 95% confidence interval 0.65-0.99)., Conclusions: This study has demonstrated, using subjective and objective measures, that preoperative BNP is a predictor of functional deterioration following lung resection. BNP may have a role in preoperative risk stratification in this population, allowing therapy in future to be targeted towards high-risk patients with the aim of preventing postoperative cardiac dysfunction., Clinical Trial Registration Number: NCT01892800., (© The Author(s) 2019. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.)

Background: Recent outbreaks of dengue and other Aedes aegypti-borne arboviruses highlight the importance of a rapid response for effective vector control. Data on insecticide resistance and underlying mechanisms are essential for outbreak preparedness, but are sparse in much of Africa. We investigated the levels and heterogeneity of insecticide resistance and mechanisms of Ae. aegypti from contrasting settings within and around Ouagadougou, Burkina Faso., Methodology/principal Findings: Bioassays were performed on larvae and adults to diagnose prevalence of resistance, and to assess levels where resistance was detected. Investigation of resistance mechanisms was performed using synergist bioassays, knockdown resistance (kdr) target site mutation genotyping and quantitative PCR expression analysis of candidate P450 genes. Larval dose-response assays indicated susceptibility to the organophosphates tested. Adult females were also susceptible to organophosphates, but resistance to carbamates was suspected in urban and semi-urban localities. Females from all localities showed resistance to pyrethroids but resistance prevalence and level were higher in urban and especially in semi-urban areas, compared to the rural population. Environment was also associated with susceptibility: adults reared from larvae collected in tires from the semi-urban site were significantly less resistant to pyrethroids than those collected from large outdoor drinking water containers ('drums'). Susceptibility to both pyrethroids tested was largely restored by pre-exposure to Piperonyl Butoxide (PBO), suggesting a strong metabolic basis to resistance. The 1534C kdr mutation was nearly fixed in semi-urban and urban areas but was far less common in the rural area, where the 1016I kdr mutation frequency was also significantly lower. P450 gene analysis detected limited over-expression of single candidates but significantly elevated average expression in the semi-urban site compared to both a susceptible laboratory colony, and females from the other collection sites., Conclusions/significance: Our results reveal pyrethroid resistance and paired kdr mutations in both urban and semi-urban sites at levels that are unprecedented for mainland Africa. The combination of target site and metabolic mechanisms is common in Ae. aegypti populations from other continents but is a worrying finding for African populations. However, organophosphate insecticides are still active against both larvae and adults of Ae. aegypti, providing useful insecticidal options for control and resistance management., Competing Interests: The authors have declared that no competing interests exist.

Background: Resistance to pyrethroid insecticides involving kdr mutations is widespread in Aedes aegypti (L.) (Diptera: Culicidae) and potentially could impact control efforts in endemic countries. Dengue cases had been sporadic in Burkina Faso for over a decade prior to the 2016-2017 outbreak that resulted in 15,074 suspected cases and 36 deaths, mainly in Ouagadougou. These outbreaks highlighted the lack of information on numerous aspects of the biology, behaviour and insecticide status of local dengue vector populations that are fundamental to vector control., Results: We investigated the insecticide resistance profiles and the kdr mutations involved in pyrethroid resistance of Ae . aegypti from Somgandé, a district of Ouagadougou. WHO bioassays revealed that the local Ae . aegypti populations were highly resistant to pyrethroids with mortalities of 15% for permethrin and 37% for deltamethrin. Resistance to carbamates was also detected with mortalities of 55% for propoxur and 90% for bendiocarb, but high mortalities (> 97%) to organophosphates (malathion and fenitrothion) indicated susceptibility. Allele-specific PCR and voltage-gated sodium channel gene sequencing showed a very high frequency (97%) of the F1534C kdr allele whilst the V1016I kdr mutation frequency was 46%. Association of dual-locus kdr mutations was detected for permethrin resistance., Conclusion: We conclude that in this locality of Burkina Faso, Ae . aegypti is resistant to pyrethroid and carbamate insecticides but remains susceptible to organophosphates, providing useful information for possible future control., Competing Interests: Not applicable.Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Background: There is a need for effective methods to control Aedes aegypti and prevent the transmission of dengue, chikungunya, yellow fever and Zika viruses. Insecticide treated screening (ITS) is a promising approach, particularly as it targets adult mosquitoes to reduce human-mosquito contact., Methodology/principal Findings: A cluster-randomised controlled trial evaluated the entomological efficacy of ITS based intervention, which consisted of the installation of pyrethroid-impregnated long-lasting insecticide-treated netting material fixed as framed screens on external doors and windows. A total of 10 treatment and 10 control clusters (100 houses/cluster) were distributed throughout the city of Merida, Mexico. Cross-sectional entomological surveys quantified indoor adult mosquito infestation at baseline (pre-intervention) and throughout four post-intervention (PI) surveys spaced at 6-month intervals corresponding to dry/rainy seasons over two years (2012-2014). A total of 844 households from intervention clusters (86% coverage) were protected with ITS at the start of the trial. Significant reductions in the indoor presence and abundance of Ae. aegypti adults (OR = 0.48 and IRR = 0.45, P<0.05 respectively) and the indoor presence and abundance of Ae. aegypti female mosquitoes (OR = 0.47 and IRR = 0.44, P<0.05 respectively) were detected in intervention clusters compared to controls. This high level of protective effect was sustained for up to 24 months PI. Insecticidal activity of the ITS material declined with time, with ~70% mortality being demonstrated in susceptible mosquito cohorts up to 24 months after installation., Conclusions/significance: The strong and sustained entomological impact observed in this study demonstrates the potential of house screening as a feasible, alternative approach to a sustained long-term impact on household infestations of Ae. aegypti. Larger trials quantifying the effectiveness of ITS on epidemiological endpoints are warranted and therefore recommended.