Vector Mosquito Species Research Articles

Ecological Niche and Positive Clusters of Two West Nile Virus Vectors in Ontario, Canada

West Nile virus (WNV) is a mosquito-borne pathogen associated with uncommon but severe neurological complications in humans, especially among the elderly and immune-compromised. In Northeastern North America, the Culex pipiens/restuans complex and Aedes vexans are the two principal vector mosquito species/species groups of WNV. Using a 10-year surveillance dataset of WNV vector captures at 118 sites across an area of 40,000 km2 in Eastern Ontario, Canada, the ecological niches of Cx. pipiens/restuans and Aedes vexans were modeled by random forest analysis. Spatiotemporal clusters of WNV-positive mosquito pools were identified using Kulldorf’s spatial scan statistic. The study region encompasses land cover types and climate representative of highly populated Southeastern Canada. We found highest vector habitat suitability in the eastern half of the study area, where temperatures are generally warmer (variable importance > 0.40) and residential and agricultural cropland cover is more prominent (variable importance > 0.25). We found spatiotemporal clusters of high WNV infection rates around the city of Ottawa in both mosquito vector species. These results support the previous literature in the same region and elsewhere suggesting areas surrounding highly populated areas are also high-risk areas for vector-borne zoonoses such as the WNV.

A Deep Transfer Learning Framework for the Multi-Class Classification of Vector Mosquito Species

A Deep Transfer Learning Framework for the Multi-Class Classification of Vector Mosquito Species

DAYTIME RESTING ACTIVITY OF AEDES AEGYPTI AND CULEX QUINQUEFASCIATUS POPULATIONS IN NORTHERN MEXICO.

Aedes aegypti and Culex quinquefasciatus are disease vectors distributed throughout much of the world and are responsible for a high burden of vector-borne disease, which has increased during the last 2 decades. Most pathogens vectored by these mosquitoes do not have therapeutic remedies; thus, combating these diseases is dependent upon vector control. Improvements in vector control strategies are urgently needed, but these hinge on understanding the biology and ecology of Ae. aegypti and Cx. quinquefasciatus. Both species have been extensively investigated, but further knowledge on diel resting activity of these vectors can improve vector surveillance and control tools for targeting resting vector populations. From April to December 2021, we determined outdoor daytime resting habits of Ae. aegypti and Cx. quinquefasciatus male, female, and blood-fed female populations in Reynosa, Mexico, using large red odor-baited wooden box traps. The daytime resting activity for Ae. aegypti males, females, and blood-fed females was restricted to a period between 0900 h and 1300 h, with a peak at 0900 h, while the resting activity of Cx. quinquefasciatus male, female, and blood-fed females was between 0700 h and 1100 h, with a peak at 0700 h. A generalized additive model was developed to relate relative humidity and temperature to resting Cx. quinquefasciatus and Ae. aegypti male, female, and blood-fed populations caught in traps. This study advances the understanding of outdoor resting behavior for 2 important vector mosquito species and discusses future studies to fill additional knowledge gaps.

Characterizing antibody responses to mosquito salivary antigens of the Southeast Asian vectors of malaria and dengue with a human challenge model of controlled exposure: a protocol.

Background: Measurement of antibody titers directed against mosquito salivary antigens in blood samples has been proposed as an outcome measure to assess human exposure to vector bites. However, only a handful of antigens have been identified and the specificity and longitudinal dynamics of antibody responses are not well known. We report the protocol of a clinical trial of controlled exposure to mosquito bites that aims to identify and validate biomarkers of exposure to bites of mosquito vector species that transmit malaria and dengue in Southeast Asia and some other parts of the world. Methods: This study is an exploratory factorial randomized control trial of controlled exposure to mosquito bites with 10 arms corresponding to different species ( Aedes aegypti, Ae. albopictus, Anopheles dirus, An. maculatus and An. minimus) and numbers of bites (35 or 305 bites in total over 6 weeks). Blood samples will be collected from study participants before, during and after mosquito biting challenges. Candidate peptides will be identified from published literature with antigen prediction algorithms using mosquito DNA sequence data and with immunoblotting assays carried out using protein extracts of dissected mosquito salivary glands and participants samples. Antibody titers against candidate peptides will be determined in participants samples with high-throughput cutting-edge immuno-assays. Quantification of the antibody response profile over time (including an estimate of the decay rate) and the effect of the number of bites on the antibody response will be determined using linear and logistic mixed-effects models for the continuous and the binary response, respectively. Conclusion: This research is expected to generate important knowledge for vector sero-surveillance and evaluation of vector-control interventions against malaria and dengue in the Greater Mekong Subregion. Registration: This study is registered with clinicaltrials.gov (NCT04478370) on July 20 th, 2020.

Using community science data to assess the association between urbanization and the presence of invasive Aedes species in Hungary

BackgroundUrbanization can be a significant contributor to the spread of invasive mosquito vector species, and the diseases they carry, as urbanized habitats provide access to a great density of food resources (humans and domestic animals) and offer abundant breeding sites for these vectors. Although anthropogenic landscapes are often associated with the presence of invasive mosquito species, we still have little understanding about the relationships between some of these and the built environment.MethodsThis study explores the association between urbanization level and the occurrence of invasive Aedes species, specifically Aedes albopictus, Aedes japonicus, and Aedes koreicus, in Hungary, using data from a community (or citizen) science program undertaken between 2019 and 2022.ResultsThe association between each of these species and urbanized landscapes within an extensive geographic area was found to differ. Using the same standardized approach, Ae. albopictus showed a statistically significant and positive relationship with urbanization, whereas Ae. japonicus and Ae. koreicus did not.ConclusionsThe findings highlight the importance of community science to mosquito research, as the data gathered using this approach can be used to make qualitative comparisons between species to explore their ecological requirements.Graphical

IN-HOUSE TESTING OF MOSQUITO POOLS FOR WEST NILE VIRUS USING COMMERCIALLY AVAILABLE IMMUNOASSAY AND REAL-TIME REVERSE-TRANSCRIPTASE POLYMERASE CHAIN REACTION KITS

Arbovirus surveillance methods are an integral part of integrated mosquito management programs, providing information on arboviral presence, location, and transmission potential. For many vector control agencies, surveillance entails collecting vector mosquito species from the field and testing representative mosquito pools using commercially available immunoassays or real-time reverse transcriptase polymerase chain reaction test kits. In 2016, the Collier Mosquito Control District established an in-house arbovirus surveillance program to screen mosquitoes for a variety of diseases, including the endemic arbovirus, West Nile virus (WNV). Although guidance on interpreting test results is provided by the manufacturer, end users of commercial test kits are encouraged to establish their own cut-off values signifying an arbovirus positive mosquito pool. Here we report the Collier Mosquito Control District’s efforts to develop cut-off values for mosquito pools using two commercially available WNV test kits.

The influence of weather on the population dynamics of common mosquito vector species in the Canadian Prairies

BackgroundMosquito seasonal activity is largely driven by weather conditions, most notably temperature, precipitation, and relative humidity. The extent by which these weather variables influence activity is intertwined with the animal’s biology and may differ by species. For mosquito vectors, changes in weather can also alter host–pathogen interactions thereby increasing or decreasing the burden of disease.MethodsIn this study, we performed weekly mosquito surveillance throughout the active season over a 2-year period in Manitoba, Canada. We then used Generalized Linear Mixed Models (GLMMs) to explore the relationships between weather variables over the preceding 2 weeks and mosquito trap counts for four of the most prevalent vector species in this region: Oc. dorsalis, Ae. vexans, Cx. tarsalis, and Cq. perturbans.ResultsMore than 265,000 mosquitoes were collected from 17 sampling sites throughout Manitoba in 2020 and 2021, with Ae. vexans the most commonly collected species followed by Cx. tarsalis. Aedes vexans favored high humidity, intermediate degree days, and low precipitation. Coquillettidia perturbans and Oc. dorsalis activity increased with high humidity and high rainfall, respectively. Culex tarsalis favored high degree days, with the relationship between number of mosquitoes captured and precipitation showing contrasting patterns between years. Minimum trapping temperature only impacted Ae. vexans and Cq. perturbans trap counts.ConclusionsThe activity of all four mosquito vectors was affected by weather conditions recorded in the 2 weeks prior to trapping, with each species favoring different conditions. Although some research has been done to explore the relationships between temperature/precipitation and Cx. tarsalis in the Canadian Prairies, to our knowledge this is the first study to investigate other commonly found vector species in this region. Overall, this study highlights how varying weather conditions can impact mosquito activity and in turn species-specific vector potential.Graphical

Geospatial modelling of lymphatic filariasis and malaria co-endemicity in Nigeria.

Lymphatic filariasis (LF) and malaria are important vector-borne diseases that are co-endemic throughout Nigeria. These infections are transmitted by the same mosquito vector species in Nigeria and their transmission is similarly influenced by climate and sociodemographic factors. The goal of this study was to assess the relationship between the geospatial distribution of both infections in Nigeria to better coordinate interventions. We used national survey data for malaria from the Demographic and Health Survey dataset and site-level LF mapping data from the Nigeria Lymphatic Filariasis Control Programme together with a suite of predictive climate and sociodemographic factors to build geospatial machine learning models. These models were then used to produce continuous gridded maps of both infections throughout Nigeria. The R2 values for the LF and malaria models were 0.68 and 0.59, respectively. Also, the correlation between pairs of observed and predicted values for LF and malaria models were 0.69 (95% confidence interval [CI] 0.61 to 0.79; p<0.001) and 0.61 (95% CI 0.52 to 0.71; p<0.001), respectively. However, we observed a very weak positive correlation between overall overlap of LF and malaria distribution in Nigeria. The reasons for this counterintuitive relationship are unclear. Differences in transmission dynamics of these parasites and vector competence may contribute to differences in the distribution of these co-endemic diseases.

Species Composition and Geographic Distribution of Culicinae Mosquitoes and Their Possible Infection with West Nile Virus in Hormozgan Province, Southern Iran.

West Nile virus (WNV) is a mosquito-borne virus that causes a febrile disease and may cause a fatal neurological illness in humans. We aimed to investigate the geographic distribution of Culicinae mosquitoes and their possible infection with WNV in Hormozgan Province, southern Iran. Field studies were carried out from June 2017 to May 2019. Different mosquito sampling methods were used monthly to collect mosquitoes from 22 sites. The Real-Time PCR technique was used to detect the virus infection in the mosquitoes. Overall, 6165 mosquitoes were caught. The species were Culex theileri (33.25%), Cx. pipiens (20.45%), Cx. quinquefasciatus (10.51%), Aedes caspius (6.33%), Cx. tritaeniorhynchus (5.82%), Ae. vexans (4.10%), Cx. sinaiticus (3.62%), Cx. antennatus (3.29%), Culiseta longiareolata (2.81%), Cx. perexiguus (2.03%), Cs. subochrea (1.95%), Cx. mimeticus (1.49%), Cx. pusillus (1.38%), Cx. univittatus (1.27%), Cx. modestus (1.14%), and Cx. sitiens (0.57%). The molecular detection of virus infection in mosquitoes found to be negative for WNV. The presence of many species of mosquito vectors and high population traffic increase the risk of disease transmission is very high. Therefore, the way to restrict WNV infection factors is increasing the knowledge for personal protection measures to prevent mosquito bites.

Mosquito community composition in Central District, Botswana: insights from a malaria endemic to non-endemic gradient

Spatial distribution of vector mosquitoes plays a critical role in the dynamics of associated diseases’ spread across diverse landscapes. In Botswana, six Districts are implicated as malaria endemic zones, one of which is the Central District comprising both malaria endemic and non-endemic sub-districts. Despite being the biggest in the country, mosquito diversity in this District is under-explored, more so in the malaria non-endemic sub-districts. Here, we thus sampled mosquito adults and larvae from the malaria endemic sub-district of Bobirwa and non-endemic sub-districts of Palapye and Serowe, to determine spatial mosquito abundance and diversity in the District. Overall, all the sub-districts had a representation of key mosquito taxa of medical and economic importance (Aedes, Culex and Anopheles), irrespective of malarial endemicity status. Bobirwa had the highest number of mosquitoes sampled (429) although the greatest species richness (0.8511) was observed from Palapye. Moreover, Palapye also recorded a species from another genus, Culiseta longiareolata, a known natural vector of avian malaria parasites. Given global climate shift projections for the region, there is a need for continuous area-wide surveillance for vector mosquitoes and associated parasites in curbing the risk of emerging and re-emerging infections. While the Anopheles-centric approach to mosquito control is still necessary, a holistic approach, incorporating other vector incriminated mosquito species is warranted, particularly given shifting climates and the presence of invasive disease associated vector mosquito species.

Characterizing antibody responses to mosquito salivary antigens of the Southeast Asian vectors of malaria and dengue with a human challenge model of controlled exposure: a protocol

Background: Measurement of antibody titers directed against mosquito salivary antigens in blood samples has been proposed as an outcome measure to assess human exposure to vector bites. However, only a handful of antigens have been identified and the specificity and longitudinal dynamics of antibody responses are not well known. We report the protocol of a clinical trial of controlled exposure to mosquito bites that aims to identify and validate biomarkers of exposure to bites of mosquito vector species that transmit malaria and dengue in Southeast Asia and some other parts of the world. Methods: This study is an exploratory factorial randomized control trial of controlled exposure to mosquito bites with 10 arms corresponding to different species (Aedes aegypt, Ae. albopictus, Anopheles dirus, An. maculatus and An. minimus) and numbers of bites (35 or 305 bites in total over 6 weeks). Blood samples will be collected from study participants before, during and after mosquito biting challenges. Candidate peptides will be identified from published literature with antigen prediction algorithms using mosquito DNA sequence data and with immunoblotting assays carried out using protein extracts of dissected mosquito salivary glands and participants samples. Antibody titers against candidate peptides will be determined in participants samples with high-throughput cutting-edge immuno-assays. Quantification of the antibody response profile over time (including an estimate of the decay rate) and the effect of the number of bites on the antibody response will be determined using linear and logistic mixed-effects models for the continuous and the binary response, respectively. Conclusion: This research is expected to generate important knowledge for vector sero-surveillance and evaluation of vector-control interventions against malaria and dengue in the Greater Mekong Subregion. Registration: This study is registered with clinicaltrials.gov (NCT04478370) on July 20th, 2020.

Mosquito-Borne Diseases and Their Control Strategies: An Overview Focused on Green Synthesized Plant-Based Metallic Nanoparticles.

Mosquitoes act as vectors of pathogens that cause most life-threatening diseases, such as malaria, Dengue, Chikungunya, Yellow fever, Zika, West Nile, Lymphatic filariasis, etc. To reduce the transmission of these mosquito-borne diseases in humans, several chemical, biological, mechanical, and pharmaceutical methods of control are used. However, these different strategies are facing important and timely challenges that include the rapid spread of highly invasive mosquitoes worldwide, the development of resistance in several mosquito species, and the recent outbreaks of novel arthropod-borne viruses (e.g., Dengue, Rift Valley fever, tick-borne encephalitis, West Nile, yellow fever, etc.). Therefore, the development of novel and effective methods of control is urgently needed to manage mosquito vectors. Adapting the principles of nanobiotechnology to mosquito vector control is one of the current approaches. As a single-step, eco-friendly, and biodegradable method that does not require the use of toxic chemicals, the green synthesis of nanoparticles using active toxic agents from plant extracts available since ancient times exhibits antagonistic responses and broad-spectrum target-specific activities against different species of vector mosquitoes. In this article, the current state of knowledge on the different mosquito control strategies in general, and on repellent and mosquitocidal plant-mediated synthesis of nanoparticles in particular, has been reviewed. By doing so, this review may open new doors for research on mosquito-borne diseases.

Spatiotemporal distribution of vector mosquito species and areas at risk for arbovirus transmission in Maricopa County, Arizona

Mosquito-borne diseases are a major global public health concern and mosquito surveillance systems are essential for the implementation of effective mosquito control strategies. The objective of our study is to determine the spatiotemporal distribution of vector mosquito species in Maricopa County, AZ from 2011 to 2021, and to identify the hotspot areas for West Nile virus (WNV) and St. Louis Encephalitis virus (SLEV) transmission in 2021. The Maricopa County Mosquito Control surveillance system utilizes BG-Sentinel and EVS-CDC traps throughout the entire urban and suburban areas of the county. We estimated specific mosquito species relative abundance per unit area using the Kernel density estimator in ArcGIS 10.2. We calculated the distance between all traps in the surveillance system and created a 4 km buffer radius around each trap to calculate the extent to which each trap deviated from the mean number of Culex quinquefasciatus and Culex tarsalis collected in 2021. Our results show that vector mosquito species are widely distributed and abundant in the urban areas of Maricopa County. A total of 691,170Cx. quinquefasciatus, 542,733 Cx. tarsalis, and 292,305 Aedes aegypti were collected from 2011 to 2022. The relative abundance of Ae. aegypti was highly seasonal peaking in the third and fourth quarters of the year. Culex quinquefasciatus, on the other hand, was abundant throughout the year with several regions consistently yielding high numbers of mosquitoes. Culex tarsalis was abundant but it only reached high numbers in well-defined areas near irrigated landscapes. We also detected high levels of heterogeneity in the risk of WNV and SLEV transmission to humans disregarding traps geographical proximity. The well-defined species-specific spatiotemporal and geographical patterns found in this study can be used to inform vector control operations.

Diel activity patterns of vector mosquito species in the urban environment: Implications for vector control strategies.

Mathematical models have been widely used to study the population dynamics of mosquitoes as well as to test and validate the effectiveness of arbovirus outbreak responses and mosquito control strategies. The objective of this study is to assess the diel activity of mosquitoes in Miami-Dade, Florida, and Brownsville, Texas, the most affected areas during the Zika outbreak in 2016-2017, and to evaluate the effectiveness of simulated adulticide treatments on local mosquito populations. To assess variations in the diel activity patterns, mosquitoes were collected hourly for 96 hours once a month from May through November 2019 in Miami-Dade County, Florida, and Brownsville, Texas. We then performed a PERMANOVA followed by a SIMPER analysis to assess whether the abundance and species richness significantly varies at different hours of the day. Finally, we used a mathematical model to simulate the population dynamics of 5 mosquito vector species and evaluate the effectiveness of the simulated adulticide applications. A total of 14,502 mosquitoes comprising 17 species were collected in Brownsville and 10,948 mosquitoes comprising 19 species were collected in Miami-Dade County. Aedes aegypti was the most common mosquito species collected every hour in both cities and peaking in abundance in the morning and the evening. Our modeling results indicate that the effectiveness of adulticide applications varied greatly depending on the hour of the treatment. In both study locations, 9 PM was the best time for adulticide applications targeting all mosquito vector species; mornings/afternoons (9 AM- 5 PM) yielded low effectiveness, especially for Culex species, while at night (12 AM- 6 AM) the effectiveness was particularly low for Aedes species. Our results indicate that the timing of adulticide spraying interventions should be carefully considered by local authorities based on the ecology of the target mosquito species in the focus area.

Entomopathogenic nematodes isolated from agricultural areas of Thailand and their activity against the larvae of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae)

Entomopathogenic nematodes (EPNs) of the genera Steinernema and Heterorhabditis have been considered to be effective biological control agents for several insects. In this study, we isolated and identified EPNs from soil samples in agricultural areas of northern Thailand and evaluated their efficacy for controlling larvae of three mosquito vector species, Aedes aegypti, Ae. albopictus and Culex quinquefasciatus. A total of 51 of 1,000 soil samples were positive (5.1% prevalence) for EPNs, which were identified through sequencing of the rDNA and ITS to 37 Steinernema isolates (3.7%) and 14 Heterorhabditis isolates (1.4%). For the bioassay, the larvae of mosquitoes were exposed to Steinernema surkhetense (eALN6.3_TH), Steinernema lamjungense (eALN11.5_TH), Heterorhabditis indica (eACM14.2_TH) and Heterorhabditis bacteriophora (eALN18.2_TH). Heterorhabditis bacteriophora showed the highest efficacy against Ae. aegypti and Cx. quinquefasciatus. At 96 h after exposure, the mortality rates were 60.0 and 91.7%, respectively. The EPNs were observed in the dead mosquito larvae, which were mostly found in the thorax followed by the head and abdomen. Some EPNs were dead with melanization, and some were able to survive in the cavity of mosquito larvae. Our results show the low prevalence of EPN in agricultural areas of Thailand. Moreover, H. bacteriophora may be considered an alternative biocontrol agent for managing and controlling these vector mosquitoes.

Phylogenetic Analysis of Wesselsbron Virus Isolated from Field-Captured Mosquitoes during a Rift Valley Fever Outbreak in Kabale District, Uganda-2016.

After confirmation of two human cases of Rift Valley fever (RVF) in March 2016 in the Kabale district of Uganda, an entomological investigation was conducted with a focus on mosquito species composition and abundance of known and potential mosquito vector species, and virus testing to identify species most likely involved in Rift Valley fever virus transmission. This information could be used to forecast risk and facilitate improvement of prevention and response tools for use in preventing or controlling future outbreaks. From these collections, two virus isolates were obtained, one each from a pool of Aedes tricholabis and Ae. gibbinsi. Next-generation sequencing identified both isolates as Wesselsbron virus, family Flaviviridae, a neglected arbovirus of economic importance. These are the first reported Wesselsbron virus isolates from Uganda since 1966.

Co-occurrence probabilities between mosquito vectors of West Nile and Eastern equine encephalitis viruses using Markov Random Fields (MRFcov)

Mosquito vectors of eastern equine encephalitis virus (EEEV) and West Nile virus (WNV) in the USA reside within broad multi-species assemblages that vary in spatial and temporal composition, relative abundances and vector competence. These variations impact the risk of pathogen transmission and the operational management of these species by local public health vector control districts. However, most models of mosquito vector dynamics focus on single species and do not account for co-occurrence probabilities between mosquito species pairs across environmental gradients. In this investigation, we use for the first time conditional Markov Random Fields (CRF) to evaluate spatial co-occurrence patterns between host-seeking mosquito vectors of EEEV and WNV around sampling sites in Manatee County, Florida. Specifically, we aimed to: (i) quantify correlations between mosquito vector species and other mosquito species; (ii) quantify correlations between mosquito vectors and landscape and climate variables; and (iii) investigate whether the strength of correlations between species pairs are conditional on landscape or climate variables. We hypothesized that either mosquito species pairs co-occur in patterns driven by the landscape and/or climate variables, or these vector species pairs are unconditionally dependent on each other regardless of the environmental variables. Our results indicated that landscape and bioclimatic covariates did not substantially improve the overall model performance and that the log abundances of the majority of WNV and EEEV vector species were positively dependent on other vector and non-vector mosquito species, unconditionally. Only five individual mosquito vectors were weakly dependent on environmental variables with one exception, Culiseta melanura, the primary vector for EEEV, which showed a strong correlation with woody wetland, precipitation seasonality and average temperature of driest quarter. Our analyses showed that majority of the studied mosquito species’ abundance and distribution are insignificantly better predicted by the biotic correlations than by environmental variables. Additionally, these mosquito vector species may be habitat generalists, as indicated by the unconditional correlation matrices between species pairs, which could have confounded our analysis, but also indicated that the approach could be operationalized to leverage species co-occurrences as indicators of vector abundances in unsampled areas, or under scenarios where environmental variables are not informative.Graphical

Efficacy of nanogels containing carvacrol, cinnamaldehyde, thymol, and a mix compared to a standard repellent against Anopheles stephensi

Mosquito-borne diseases are responsible for about one million death annually worldwide. Especially in endemic regions, using repellents is a practical approach to preventing such diseases. In this study, nanoemulsions containing carvacrol, cinnamaldehyde, thymol, and a mix of them were first prepared. The nanoemulsions were then gellified by adding carboxymethyl cellulose (CMC). Besides, viscosity and successful loading were investigated using viscometry and Attenuated Total Reflection-Fourier Transform InfraRed (ATR-FTIR) analyses. After that, the complete protection time of the nanogels compared to DEET (N, N-diethyl-meta-toluamide) was evaluated against the Anopheles stephensi. The repellent activity of commercial DEET (40% w/v) was 480 ± 30 min; interestingly, nanogel-containing carvacrol (2.4% w/v) showed 470 ± 17 min protection time. Besides, no significant difference was observed between nanogels containing cinnamaldehyde and a mix of them and DEET. The straightforward, repeatable preparation method and no need for advanced devices are some advantages of the developed nanogels. Results suggested that mentioned nanogels, especially carvacrol, could be considered novel natural repellents against A. stephensi and other species of vector mosquitoes.

Clock genes regulate mating activity rhythms in the vector mosquitoes, Aedes albopictus and Culex quinquefasciatus.

Endogenous circadian rhythms result from genetically-encoded molecular clocks, whose components and downstream output factors cooperate to generate cyclic changes in activity. Mating is an important activity of mosquitoes, however, the key aspects of mating rhythm patterns and their regulatory mechanisms in two vector mosquito species, Aedes albopictus and Culex quinquefasciatus, remain unclear. We determined and compared the diel mating activity rhythms of these two mosquito species and discovered that Ae. albopictus had mating peaks in the light/dark transition periods (ZT0-3 and ZT9-12), while Cx. quinquefasciatus only had a mating peak at ZT12-15. Knockouts of the clock (clk) orthologous genes (Aalclk and Cxqclk) resulted in phase delay or phase reversal of the mating peaks in Ae. albopictus and Cx. quinquefasciatus, respectively. In addition, the temporal expression pattern of the desaturase orthologous genes, desat1, in both mosquito species was also different in respective wild-type strains and showed phase changes similar to the mating rhythms in clk mutant strains. Inhibition of desat1 expression resulted in decreased mating activity in male mosquitoes of both species but not females. In addition, desat1 regulated cuticular hydrocarbons' synthesis in both species. Silencing desat1 in male Ae. albopictus resulted in decreases of nonadecane and tricosane, which promoted mating, with concomitant increases of heptacosane, which inhibited mating. Silencing desat1 in male Cx. quinquefasciatus also resulted in decreases of tricosane, which promoted mating. These results suggest that Aalclk and Cxqclk have significant roles in the mating activity rhythms in both Ae. albopictus and Cx. quinquefasciatus by regulating the temporal expression of the desat1 gene under LD cycles, which affects sex pheromone synthesis and mating. This work provides insights into the molecular regulatory mechanism of distinct mating rhythm of Ae. albopictus and Cx. quinquefasciatus and may provide a basis for the control of these two important vector mosquitoes.

Novel genome sequences and evolutionary dynamics of the North American anopheline species Anopheles freeborni, Anopheles crucians, Anopheles quadrimaculatus, and Anopheles albimanus.

Anopheles mosquitoes are the principal vectors for malaria and lymphatic filariasis, and evidence for arboviral transmission under laboratory and natural contexts has been demonstrated. Vector management approaches require an understanding of the ecological, epidemiological, and biological contexts of the species in question, and increased interest in gene drive systems for vector control applications has resulted in an increased need for genome assemblies from understudied mosquito vector species. In this study, we present novel genome assemblies for Anopheles crucians, Anopheles freeborni, Anopheles albimanus, and Anopheles quadrimaculatus and examine the evolutionary relationship between these species. We identified 790 shared single-copy orthologs between the newly sequenced genomes and created a phylogeny using 673 of the orthologs, identifying 289 orthologs with evidence for positive selection on at least 1 branch of the phylogeny. Gene ontology terms such as calcium ion signaling, histone binding, and protein acetylation identified as being biased in the set of selected genes. These novel genome sequences will be useful in developing our understanding of the diverse biological traits that drive vectorial capacity in anophelines.