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Abstract BackgroundInsecticide resistance in malaria vectors can be spatially highly heterogeneous, yet population structure analyses frequently find relatively high levels of gene flow among mosquito populations. Few studies have contemporaneously assessed phenotypic, genotypic and population structure analysis on mosquito populations and none at fine geographical scales. In this study, genetic diversity, population structure, and insecticide resistance profiles ofAnopheles funestusandAnopheles arabiensiswere examined across mosquito populations from and within neighbouring villages. MethodsMosquitoes were collected from 11 towns in southern Mozambique, as well as from different neighbourhoods within the town of Palmeira, during the peak malaria transmission season in 2016. CDC bottle bioassay and PCR assays were performed withAnophelesmosquitoes at each site to determine phenotypic and molecular insecticide resistance profiles, respectively. Microsatellite analysis was conducted on a subsample of mosquitoes to estimate genetic diversity and population structure. ResultsPhenotypic insecticide resistance to deltamethrin was observed inAn. funestussensu stricto (s.s.) throughout the area, though a high level of mortality variation was seen. However, 98% ofAn. funestus s.s.wereCYP6P9ahomozygous resistant.An. arabiensiswas phenotypically susceptible to deltamethrin and 99% werekdrhomozygous susceptible. BothAnophelesspecies exhibited high allelic richness and heterozygosity. Significant deviations from Hardy–Weinberg equilibrium were observed, and high linkage disequilibrium was seen forAn. funestus s.s.,supporting population subdivision. However, the F_STvalues were low for both anophelines (− 0.00457 to 0.04213), N_mvalues were high (9.4–71.8 migrants per generation), AMOVA results showed almost 100% genetic variation among and within individuals, andStructureanalysis showed no clustering ofAn. funestus s.s.andAn. arabiensispopulations. These results suggest high gene flow among mosquito populations. ConclusionDespite a relatively high level of phenotypic variation in theAn. funestuspopulation, molecular analysis shows the population is admixed. These data indicate thatCYP6P9aresistance markers do not capture all phenotypic variation in the area, but also that resistance genes of high impact are likely to easily spread in the area. Conversely, other strategies, such as transgenic mosquito release programmes will likely not face challenges in this locality. 

Abstract BackgroundInsecticide resistance remains a major public health problem. Resistance surveillance is critical for effective vector control and resistance management planning. Commonly used insecticide susceptibility bioassays for mosquitoes are the CDC bottle bioassay and the WHO tube test. Less commonly used in the field but considered the gold standard for assessing insecticide susceptibility in the development of novel insecticides is the topical application bioassay. Each of these bioassays has critical differences in how they assess insecticide susceptibility that impacts their ability to differentiate between resistant and susceptible populations or determine different levels of resistance intensity. MethodsWe compared the CDC bottle bioassay, the WHO tube test, and the topical application bioassay in establishing the dose–response against deltamethrin (DM) using the DM-resistantAedes aegyptistrain MC1. Mosquitoes were exposed to a range of insecticide concentrations to establish a dose–response curve and assess variation around model predictions. In addition, 10 replicates of 20–25 mosquitoes were exposed to a fixed dose with intermediate mortality to assess the degree of variation in mortality. ResultsThe topical application bioassay exhibited the lowest amount of variation in the dose–response data, followed by the WHO tube test. The CDC bottle bioassay had the highest level of variation. In the fixed-dose experiment, a higher variance was similarly found for the CDC bottle bioassay compared with the WHO tube test and topical application bioassay. ConclusionThese data suggest that the CDC bottle bioassay has the lowest power and the topical application bioassay the highest power to differentiate between resistant and susceptible populations and assess changes over time and between populations. This observation has significant implications for the interpretation of surveillance results from different assays. Ultimately, it will be important to discuss optimal insecticide resistance surveillance tools in terms of the surveillance objective, practicality in the field, and accuracy of the tool to reach that objective. Graphical Abstract 

West Nile virus (WNV) is the leading mosquito-borne disease causing-pathogen in the United States. Concerningly, there are no prophylactics or drug treatments for WNV and public health programs rely heavily on vector control efforts to lessen disease incidence. Insecticides can be effective in reducing vector numbers if implemented strategically, but can diminish in efficacy and promote insecticide resistance otherwise. Vector control programs which employ mass-fogging applications of insecticides, often conduct these methods during the late-night hours, when diel temperatures are coldest, and without a-priori knowledge on daily mosquito activity patterns. This study’s aims were to 1) quantify the effect of temperature on the toxicity of two conventional insecticides used in fogging applications (malathion and deltamethrin) toCulex tarsalis, an important WNV vector, and 2) quantify the time of host-seeking ofCx.tarsalisand other local mosquito species in Maricopa County, Arizona. The temperature-toxicity relationship of insecticides was assessed using the WHO tube bioassay, and adultCx.tarsalis, collected as larvae, were exposed to three different insecticide doses at three temperature regimes (15, 25, and 35°C; 80% RH). Time of host-seeking was assessed using collection bottle rotators with encephalitis vector survey traps baited with dry ice, first at 3h intervals during a full day, followed by 1h intervals during the night-time. Malathion became less toxic at cooler temperatures at all doses, while deltamethrin was less toxic at cooler temperatures at the low dose. Regarding time of host-seeking,Cx.tarsalis,Aedes vexans, andCulex quinquefasciatuswere the most abundant vectors captured. During the 3-hour interval surveillance over a full day,Cx.tarsaliswere most-active during post-midnight biting (00:00–6:00), accounting for 69.0% of allCx.tarsalis, while pre-midnight biting (18:00–24:00) accounted for 30.0% ofCx.tarsalis. During the 1-hour interval surveillance overnight,Cx.tarsaliswere most-active during pre-midnight hours (18:00–24:00), accounting for 50.2% ofCx.tarsaliscaptures, while post-midnight biting (00:00–6:00) accounted for 49.8% ofCx.tarsalis. Our results suggest that programs employing large-scale applications of insecticidal fogging should consider temperature-toxicity relationships coupled with time of host-seeking data to maximize the efficacy of vector control interventions in reducing mosquito-borne disease burden. 

Arizona is home to many mosquito species, some of which are known vectors of infectious diseases that harm both humans and animals. Here, we provide an overview of the 56 mosquito species that have been identified in the State to date, but also discuss their known feeding preference and the diseases they can (potentially) transmit to humans and animals. This list is unlikely to be complete for several reasons: (i) Arizona’s mosquitoes are not systematically surveyed in many areas, (ii) surveillance efforts often target specific species of interest, and (iii) doubts have been raised by one or more scientists about the accuracy of some collection records, which has been noted in this article. There needs to be an integrated and multifaceted surveillance approach that involves entomologists and epidemiologists, but also social scientists, wildlife ecologists, ornithologists, representatives from the agricultural department, and irrigation and drainage districts. This will allow public health officials to (i) monitor changes in current mosquito species diversity and abundance, (ii) monitor the introduction of new or invasive species, (iii) identify locations or specific populations that are more at risk for mosquito-borne diseases, and (iv) effectively guide vector control.