A numbers game: mosquito-based arbovirus surveillance in two distinct geographic regions of Latin America.

Aedes mosquitoes, as vectors of medically important arthropod-borne viruses (arboviruses), constitute a major public health threat that requires entomological and epidemiological surveillance to guide vector control programs to prevent and reduce disease transmission. In this study, we present the collaborative effort of 1 year of Aedes ageypti (Linnaeus, 1762) mosquito-based arbovirus surveillance in 2 geographically distinct regions of Latin America (Nicaragua and Ecuador). Adult female mosquitoes were collected using backpack aspirators in over 2,800 randomly selected households (Nicaragua, Ecuador) and 100 key sites (Nicaragua) from 8 distinct communities (Nicaragua: 2, Ecuador: 6). A total of 1,358 mosquito female pools were processed for RNA extraction and viral RNA detection using real-time reverse transcription-polymerase chain reaction. Ten positive dengue virus (DENV) pools were detected (3 in Nicaragua and 7 in Ecuador), all of which were found during the rainy season and matched the serotypes found in humans (Nicaragua: DENV-1 and DENV-4; Ecuador: DENV-2). Infection rates ranged from 1.13 to 23.13, with the Nicaraguan communities having the lowest infection rates. Our results demonstrate the feasibility of detecting DENV-infected Aedes mosquitoes in low-resource settings and underscore the need for targeted mosquito arbovirus sampling and testing, providing valuable insights for future surveillance programs in the Latin American region.

Investigating knowledge, attitude, and self-care behavior concerning Aedes mosquito bites and the knowledge of dengue fever among Hormozgan residents in the south of Iran.

BACKGROUND: In recent years, with the identification of the Aedes mosquito as a vector of dengue fever (DF) in Hormozgan province, this southern province was recognized nationwide at a great risk of DF. The present study was conducted on the Aedes mosquito prevention and DF knowledge in this province. MATERIALS AND METHODS: An online survey was conducted using a self-administered questionnaire for 1 month in 2022. The sampling method was snowball. A total number of 4,004 questionnaires were completed online by the residents of Hormozgan province. Finally, the data were analyzed using descriptive statistics, statistical tests, and statistical software SPSS (Statistical Package for the Social Science) version 22. RESULTS: The level of knowledge, attitude, and behavior concerning the self-care behavior of Aedes mosquito bites was as follows. Most participants were knowledgeable and correctly answered the questions related to the preventive ways of Aedes mosquito bites. The average rate of correct answers was 94.5%. They also mostly held a positive attitude. Regarding the attitude items, most participants (more than 85%) strongly agreed and agreed with the statements. Regarding the behavioral items, most participants (70.4%) stated they showed the aforementioned preventive behaviors to impede Aedes mosquito bites. As for the state of participants' knowledge of how the DF is transmitted, overall, in the nine examined items, on average, 40.6% answered "I do not know" and "false". CONCLUSION: As the education level of the participants showed, they did not know adequately about the DF. Judging the state of knowledge, attitude, and behavior of the less privileged and less educated social classes in Hormozgan province requires further investigation of the public living in this province after some time passes since the implementation of educational and preventive interventions. It is also necessary to enrich population-based educational interventions aiming to improve knowledge and preventive behaviors of the DF and Aedes mosquito.

Intra-species quantification reveals differences in activity and sleep levels in the yellow fever mosquito, Aedes aegypti.

Aedes aegypti is an important mosquito vector of human disease with a wide distribution across the globe. Climatic conditions and ecological pressure drive differences in the biology of several populations of this mosquito species, including blood-feeding behaviour and vector competence. However, no study has compared activity and/or sleep among different populations/lineages of Ae. aegypti. Having recently established sleep-like states in three mosquito species with observable differences in timing and amount of sleep among species, we investigated differences in activity and sleep levels among 17 Ae. aegypti lines drawn from both its native range in Africa and its invasive range across the global tropics. Activity monitoring indicates that all the lines show consistent diurnal activity, but significant differences in activity level, sleep amount, number of sleep bouts and bout duration were observed among the lines. The variation in day activity was associated with differences in host preference and ancestry for the lineages collected in Africa. This study provides evidence that the diurnal sleep and activity profiles for Ae. aegypti are consistent, but there are significant population differences for Ae. aegypti sleep and activity levels and interactions with host species may significantly impact mosquito activity.

Automated classification of mixed populations of Aedes aegypti and Culex quinquefasciatus mosquitoes under field conditions.

BACKGROUND: The recent rise in the transmission of mosquito-borne diseases such as dengue virus (DENV), Zika (ZIKV), chikungunya (CHIKV), Oropouche (OROV), and West Nile (WNV) is a major concern for public health managers worldwide. Emerging technologies for automated remote mosquito classification can be supplemented to improve surveillance systems and provide valuable information regarding mosquito vector catches in real time. METHODS: We coupled an optical sensor to the entrance of a standard mosquito suction trap (BG-Mosquitaire) to record 9151 insect flights in two Brazilian cities: Rio de Janeiro and Brasilia. The traps and sensors remained in the field for approximately 1 year. A total of 1383 mosquito flights were recorded from the target species: Aedes aegypti and Culex quinquefasciatus. Mosquito classification was based on previous models developed and trained using European populations of Aedes albopictus and Culex pipiens. RESULTS: The VECTRACK sensor was able to discriminate the target mosquitoes (Aedes and Culex genera) from non-target insects with an accuracy of 99.8%. Considering only mosquito vectors, the classification between Aedes and Culex achieved an accuracy of 93.7%. The sex classification worked better for Cx. quinquefasciatus (accuracy: 95%; specificity: 95.3%) than for Ae. aegypti (accuracy: 92.1%; specificity: 88.4%). CONCLUSIONS: The data reported herein show high accuracy, sensitivity, specificity and precision of an automated optical sensor in classifying target mosquito species, genus and sex. Similar results were obtained in two different Brazilian cities, suggesting high reliability of our findings. Surprisingly, the model developed for European populations of Ae. albopictus worked well for Brazilian Ae. aegypti populations, and the model developed and trained for Cx. pipiens was able to classify Brazilian Cx. quinquefasciatus populations. Our findings suggest this optical sensor can be integrated into mosquito surveillance methods and generate accurate automatic real-time monitoring of medically relevant mosquito species.

Impact of storm drains on the maintenance of dengue endemicity in Presidente Prudente, São Paulo, Brazil: a geospatial and epidemiologic approach.

Introduction: Dengue is a public health challenge worldwide. Brazil registered about 70% of cases in Latin America in 2023; in 2024, the country is experiencing an unprecedented increase in the number of infected individuals. By May 2024, more than 4 million people were infected. Our goal was to: (1) determine the epidemiology of dengue cases and their spatiotemporal distribution and (2) carry out a survey of the storm drains and through a geospatial analysis to determine their possible correlation with cases of dengue in Presidente Prudente, São Paulo, Brazil. Methods: Cases and information on the habitat of mosquito in the storm drain underground drainage system from 2020 to 2021 were obtained from public agencies. Larvae, pupae, and Ae. aegypti were identified according to species and described in taxonomic keys. Kernel density maps were constructed. Results: From 1996 to 2023, the prevalence of cases peaked in 2016 and 2019, and in 2023 reached alarming levels, and the city was considered hyperendemic. In 2021, 2,609 cases were registered with 2 clusters of high density. Of 5,492 storm drains analyzed, 18.0% were found to have water, 9.0% had larvae or pupae of Aedes aegypti and 91.0% were classified as dirty or damaged. A direct correlation between the kernel layer of cases in 2021 with the kernel layer of storm drains containing water (r = 0.651) and larvae and pupae (r = 0.576) was found, suggesting that storm drains are risk factors and have an impact on the maintenance of dengue endemicity. The high number of damaged units found demonstrated the lack of storm drain management, compromising the urban drainage system and possibly contributing to dengue outbreaks. Conclusion: Policymakers may use these findings to improve existing dengue control strategies focusing on the control of storm drains and increase local and global perspectives on reducing dengue outbreaks.

Aedes albopictus colonies from different geographic origins differ in their sleep and activity levels but not in the time of peak activity.

Mosquitoes occupy a wide range of habitats where they experience various environmental conditions. The ability of some species, such as the tiger mosquito, Aedes albopictus, to adapt to local conditions certainly contributes to their invasive success. Among traits that remain to be examined, mosquitoes' ability to time their activity with that of the local host population has been suggested to be of significant epidemiological importance. However, whether different populations display heritable differences in their chronotype has not been examined. Here, we compared laboratory strains originating from eight populations from three continents, monitored their spontaneous locomotor activity patterns and analysed their sleep-like states. Overall, all strains showed conserved diurnal activity concentrated in the hours preceding the crepuscule. Similarly, they all showed increased sleep levels during the morning and night hours. However, we observed strain-specific differences in the activity levels at each phase of the day. We also observed differences in the fraction of time that each strain spends in a sleep-like state, explained by variations in the sleep architecture across strains. Human population density and the latitude of the site of the geographic origin of the tested strain showed significant effects on sleep and activity patterns. Altogether, these results suggest that Ae. albopictus mosquitoes adapt to local environmental conditions via heritable adaptations of their chronotype.

Resistance of Aedes albopictus (Diptera: Culicidae) larvae and adults to insecticides based on bioassays and knockdown resistance (kdr) mutations in Zhejiang Province, China.

Aedes albopictus, a common mosquito in Zhejiang Province, is a carrier of more than twenty arboviruses. There are dozens or even hundreds of imported cases of dengue fever every year in Zhejiang Province, and there have also been many local outbreaks caused by imported cases of dengue fever. The objectives were to assess the resistance of larvae and adults of several Ae. albopictus strains in Zhejiang Province to commonly used pyrethroid insecticides (beta-cypermethrin, deltamethrin and permethrin), and detect mutations in the sodium channel gene, to further analyse the relationship between phenotypic resistance and the frequency of mutations. The resistance of eight field strains of Ae. albopictus larvae to beta-cypermethrin, deltamethrin and permethrin ranged from 8.17 to 36.06, 12.12-107.3 and 1.55-81.9, respectively, and there was a significant positive correlation of interaction resistance among the three insecticides. The mutation frequencies of I1532T and F1534S in the larvae of Ae. albopictus were 0-6.25 % and 42.19-100.00 %. Moreover, the diagnostic doses of the three pyrethroids for adult Ae. albopictus mosquitoes were 0.2510 g/L, 0.1562 g/L, and 0.9072 g/L. Except for the Zhoushan strain, which was suspected to be resistant to beta-cypermethrin, the other field strains were resistant to the three pyrethroids, and there was a significant positive correlation of cross-resistance among the three insecticides. The mutation frequencies of I1532T and F1534S of adult Ae. albopictus were 0-1.56 % and 62.50-100.00 %. In addition, the LC50 of the larvae and the mortality rate of adult Ae. albopictus after treatment with the three pyrethroids were significantly and positively correlated with the frequency of the F1534S mutation. F1534S mutation occurred earlier than I1532T mutation in both larvae and adult Ae. albopictus. F1534S mutation in the sodium channel gene may be a particular biomolecular detection marker for resistance to pyrethroid insecticides in Ae. albopictus in Zhejiang Province.

Spatial heterogeneity in the potential distribution of Aedes mosquitoes in India under current and future climatic scenarios.

Aedes is the most globally distributed mosquito genus in the 21st century and transmits various arboviral diseases. The rapid expansion of Ae. Aegypti and Ae. albopictus breeding habitats is a significant threat to global public health, driven by temperature and precipitation changes. In this study, bioclimatic variables were employed to predict the spatial distribution of Ae. aegypti and Ae. albopictus in India. The reference coordinate points of (n = 583) Aedes occurrences at a scale of ∼1 km and nineteen bioclimatic factors were retrieved to train SDM (Species Distribution Models) for both species. Maximum entropy modelling was used to predict the species' fundamental climatic niche distributions. Future projections were made using global climate models for 2021-2040 and 2081-2100 separately. The models performed reasonably well (AUC > 0.77). Both species thrived in reduced diurnal temperature and higher annual mean temperatures, with suitability increasing alongside precipitation. Ae. aegypti's projected present and future distribution was broader than that of Ae. Albopictus. The expansion of Aedes suitability varied under different future climatic scenarios. Suitability for Ae. aegypti could expand from between 17.6 and 41.1 % in 2100 under SSP (shared socioeconomic pathways) scenarios 1 and 3, respectively, whereas for Ae. albopictus suitability increased from between 10.2 and 25 % under SSP scenarios 1 and 3 respectively. Preparing for future epidemics and outbreaks requires robust vector distribution models to identify high-risk areas, allocate resources for surveillance and control, and implement prevention strategies.

Climate change impacts on dengue transmission areas in Espírito Santo state, Brazil.

Espírito Santo state, in Brazil, is a dengue-endemic region predicted to suffer from an increase in temperature and drought due to climate change, which could affect the areas with active dengue virus transmission. The study objective was modeling climatic factors and climate change effects in zones suitable for dengue virus transmission in Espírito Santo state, Brazil. Data on dengue reports from 2022 were used to determine climatic variables related to spatial distribution. The climate change projections were generated for the 2030s, 2050s, 2070s, and 2090s for three distinct Shared Socioeconomic Pathways: SSP1-2.6, SSP2-4.5 and SSP5-8.5. A maximum entropy algorithm was used to construct the three models and projections, and the results were used to calculate the ensemble mean. Isothermality, the maximum temperature of the warmest month, precipitation of the wettest month, precipitation of the warmest quarter, and annual precipitation impacted the model. Projections indicated a change in areas suitable for dengue virus transmission, varying from -30.44% in the 2070s (SSP1-2.6) to +13.07% in the 2070s (SSP5-8.5) compared to 2022. The coastal regions were consistently suitable in all scenarios. Urbanized and highly populated areas were predicted to persist with active dengue transmission in Espírito Santo state, posing challenges for public health response.

Evolutionary adaptation under climate change: Aedes sp. demonstrates potential to adapt to warming.

Climate warming is expected to shift the distributions of mosquitoes and mosquito-borne diseases, facilitating expansions at cool range edges and contractions at warm range edges. However, whether mosquito populations could maintain their warm edges through evolutionary adaptation remains unknown. Here, we investigate the potential for thermal adaptation in Aedes sierrensis, a congener of the major disease vector species that experiences large thermal gradients in its native range, by assaying tolerance to prolonged and acute heat exposure, and its genetic basis in a diverse, field-derived population. We found pervasive evidence of heritable genetic variation in acute heat tolerance, which phenotypically trades off with tolerance to prolonged heat exposure. A simple evolutionary model based on our data shows that the estimated maximum rate of evolutionary adaptation in mosquito heat tolerance typically exceeds that of projected climate warming under idealized conditions. Our findings indicate that natural mosquito populations may have the potential to track projected warming via genetic adaptation. Prior climate-based projections may thus underestimate the range of mosquito and mosquito-borne disease distributions under future climate conditions.

The spread of the invasive mosquito Aedes albopictus (Diptera: Culicidae) in Portugal: a first genetic analysis.

BACKGROUND: Aedes albopictus, commonly known as the Asian tiger mosquito, has become one of the most invasive mosquito species. Over the last 5 decades, it has been introduced and established in various tropical and temperate regions worldwide. First reported in Europe in 1979 in Albania and later in Italy in 1990, the species is now established in 13 European Union (EU)/European Economic Area (EEA) countries and 337 regions (2023). In Portugal, Ae. albopictus was first detected in the Algarve and Penafiel regions in 2017, followed by Alentejo in 2022 and Lisbon in 2023. This mosquito species poses a significant public health risk as a vector for numerous pathogenic viruses, including dengue, Zika, and chikungunya. METHODS: Aedes albopictus collected in Lisbon in 2023 were analyzed using cytochrome c oxidase I (COX) gene sequencing to understand their genetic relationships. RESULTS: Our data indicate that the Ae. albopictus mosquito populations detected in three locations in Lisbon in 2023 correspond to recent but distinct introduction events. CONCLUSIONS: Although there has been no local transmission of Aedes-transmitted viruses in mainland Portugal to date, the spread of the mosquito and increased international travel increase the risk of Aedes-borne disease outbreaks. The ongoing spread of Ae. albopictus in the country and the confirmed multiple introductions in new locations raise awareness of the need to monitor mosquito vectors to control and prevent autochthonous Aedes-borne disease outbreaks.

Bioassay-guided isolation of insect repellent compounds from Ligusticum porteri root extract.

In our program of screening natural products against the pests of medical and veterinary importance, ethanolic extract of the roots of Ligusticum porteri J.M.Coult. & Rose showed significant repellency against mosquitoes. The extract was then fractionated to test different fractions to identify the active repellent compounds. This testing resulted in the isolation of different compounds including (Z)-3-butylidenephthalide, (E)-3-butylidenephthalide, and a mixture of (E/Z)-3-butylidenephthalide. Biting deterrence of all these compounds was similar to N,N-diethyl-3-methylbenzamide (DEET) against Aedes aegypti (L.) in Klun and Debboun (K & D) bioassay. (E/Z)-3-butylidenephthalide which is a mixture of the two compounds was further tested in Ali & Khan (A & K) bioassay. Based on these data repellency of this compound was similar whereas the MED values of the mixtures of (E/Z)-3-butylidenephthalide with carotol were lower (6.25 + 6.25 = 12.5 µg/cm2) than individual treatments (25 µg/cm2). In in vivo (direct skin application bioassay), (E/Z)-3-butylidenephthalide showed excellent repellency. The residual repellency of (E/Z)-3-butylidenephthalide at 8 and 16 % application rates was 4.5 and 10-h respectively which was equal to or better than DEET with the residual time of 5 and 9-h, respectively. The mixture of (E/Z)-3-butylidenephthalide with carotol (8 + 8 %) increased the residual repellency by 2-h (44 %) as compared to (E/Z)-3-butylidenephthalide alone at a dose of 8 %. These data indicated that (E/Z)-3-butylidenephthalide is an effective mosquito repellent that is stable and has a long shelf life. The activity of this compound is extraordinary and residual time is comparable to DEET. In vivo data demonstrated an enormous potential of (E/Z)-3-butylidenephthalide as a repellent that can be developed for commercial use. However, (E/Z)-3-butylidenephthalate was found in lower amounts of the L. porteri essential oil.

Insights from multigene analysis: first report of a Southeast Asian Mosquito, Aedes (Mucidus) laniger (Diptera: Culicidae) on Jeju Island from Korea.

BACKGROUND: Certain mosquitoes are known as dominant vectors worldwide, and transmit infectious diseases. The expansion of mosquito habitats due to climate change and increased human activities poses a significant health threat by facilitating the spread of various non-native infectious diseases. This study focused on the detection of the Southeast Asian mosquito species, Aedes (Mucidus) laniger (Wiedemann, 1820) on Jeju Island, the southernmost region of the Republic of Korea (ROK), highlighting the potential risks associated with the spread of vector-borne diseases, particularly emphasizing the elevated likelihood of invasion by Southeast Asian mosquitoes. METHODS: Field surveys were conducted in August 2023 on Jeju Island. Adult mosquitoes were collected using BG-sentinel traps and identified to the species level using taxonomic keys. Morphological and molecular analyses were employed to confirm species designations. Molecular data, including mitochondrial and nuclear genes, were used for phylogenetic analysis, which was performed to compare and identify among recorded subgenera in ROK. Species distribution modeling for Ae. laniger was performed to predict potential habitats using R package 'BIOMOD2'. RESULTS: The two specimens of Ae. laniger were collected for the first time on Jeju Island. Morphological and molecular analyses confirmed the identity of this species within the subgenus Mucidus and validated the first record of this species in the ROK. We employed a simple multigene phylogenetic analysis to confirm a new mosquito record at the genus and subgenus levels, finally validating the consistency between morphological identification and molecular phylogenetic outcomes. Furthermore, we have updated the taxonomic keys for the genus Aedes in the ROK, and revised mosquito lists for Jeju Island, incorporating the inclusion of Ae. laniger. On the basis of species distribution modeling, the area of suitable habitat for Ae. laniger is expected to expand due to climate change, but this change did not appear to be meaningful in East Asia. CONCLUSIONS: This case offers the first report of the Southeast Asian mosquito, Ae. laniger, in the ROK. The detection of this species on Jeju Island suggests the potential establishment of a breeding population their habitat and raises concerns about further expansion into the Korean Peninsula. Considering the annual occurrence of mosquito-borne disease cases in the Southeast Asia, it is essential to conduct monitoring not only in Jeju Island, where Ae. laniger has been identified, but also across the entire Korean Peninsula.

Evaluation of developmental milestones and of brain measurements in rats exposed to the pesticide pyriproxyfen in prenatal period.

Pyriproxyfen is a pesticide used in Brazil to control the Aedes aegypti mosquito, vector of arboviruses like Zika and dengue. However, this pesticide is structurally similar to retinoic acid, a metabolite of vitamin A that regulates neuronal differentiation and hindbrain development during the embryonic period. Due to the similarity between pyriproxyfen and retinoic acid, studies indicate that this pesticide may have cross-reactivity with retinoid receptors. Thus, pregnant exposure to pyriproxyfen could interfere in the nervous system development of the fetal. In this context, the present study evaluated whether prenatal exposure to pyriproxyfen affects neonatal development and brain structure in rats. Wistar rat pups were divided in three experimental groups: (1) negative control (CT-)-offspring of rats that drink potable water during pregnancy; (2) pyriproxyfen (PIR)-offspring of rats exposed to Sumilarv® prenatally, a pesticide that has pyriproxyfen as active ingredient; and (3) positive control (CT+)-offspring of rats exposed to an excess of vitamin A prenatally. Only vitamin A treated-pregnant showed lower weight gain, but gestation length was similar among pregnant that received potable water, water containing vitamin A and water containing Sumilarv. In relation to the offspring, PIR group exhibits a delayed front-limb suspension response but performed early the negative geotaxis reflex. On the other hand, CT+ group exhibited lower body weight in the 1st postnatal day, delayed audio startle response, but performed early the eyelids opening and hindlimb placing response. A reduction in the maximum brain width was observed both in PIR and CT+ groups, but a reduction in the number of neurons in the M1 cortex was showed only in CT+ group. The number of glial cells in this brain area was similar between the three experimental groups studied. Although prenatal exposure to pyriproxyfen did not alter neonatal milestones in the same way as vitamin A in excess, both substances caused a reduction in the maximum width of the brain, suggesting that this pesticide can produce neurotoxic effects during the embryonic period.

Larvicidal potential of Trachyspermum ammi essential oil and Delphinium speciosum extract against malaria, dengue, and filariasis mosquito vectors.

Mosquito-borne diseases, such as malaria, dengue, and Zika, pose major public health challenges globally, affecting millions of people. The growing resistance of mosquito populations to synthetic insecticides underscores the critical need for effective and environmentally friendly larvicides. Although chemical pesticides can initially be effective, they often lead to negative environmental consequences and health hazards for non-target species, including humans. This study aimed to evaluate the larvicidal effects of Trachyspermum ammi essential oil and Delphinium speciosum extract on the larvae of three major mosquito species: Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. Mosquito larvae of Ae. aegypti, An. stephensi, and Cx. quinquefasciatus were reared under controlled laboratory conditions. The larvicidal activity of T. ammi essential oil and D. speciosum extract was evaluated through standard bioassays, using various concentrations of essential oils (10, 20, 40, 80, and 160 ppm) and extracts (160, 320, 640, 1280, and 2560 ppm) to determine the lethal concentration (LC50) values after 24 h of exposure. Fresh plant materials were collected, with the essential oil extracted via hydro-distillation, and the extract prepared using methanol solvent extraction. The chemical composition of T. ammi essential oil was examined using gas chromatography-mass spectrometry (GC-MS). Additionally, the preliminary analysis of the chemical compounds in D. speciosum extract was carried out using thin layer chromatography (TLC) and nuclear magnetic resonance spectroscopy (NMR) techniques. The results indicated that the essential oil of T. ammi exhibited more effective larvicidal activity compared to the D. speciosum extract. Specifically, the essential oil demonstrated LC50 values of 18 ppm for Cx. quinquefasciatus and 19 ppm for Ae. aegypti. In contrast, the D. speciosum extract showed the strongest larvicidal effect against An. stephensi, with an LC50 of 517 ppm. Concentrations of 40 ppm of the essential oil and 1280 ppm of the extract resulted in 100% mortality across all three species. Both the essential oil of T. ammi and the D. speciosum extract exhibited concentration-dependent larvicidal activity, and these results were statistically significant (p < 0.001) compared to the no-treatment group. GC-MS analysis revealed thymol (88.95%), o-cymen-5-ol (4.11%), and γ-terpinene (2.10%) as the major constituents of the T. ammi essential oil. Additionally, TLC verified the presence of alkaloids in both chloroform and methanolic extracts. Proton NMR identified a diterpene structure for these alkaloids. These findings suggest that T. ammi essential oil is a promising candidate for natural mosquito control strategies. Given its efficacy, further research is warranted to explore its potential in integrated vector management programs.

Characterization of the microbiome of Aedes albopictus populations in different habitats from Spain and São Tomé.

The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes.

Identification and functional analysis of C-type lectin from mosquito Aedes albopictus in response to dengue virus infection.

BACKGROUND: C-type lectins (CTLs) are a large family of proteins with sugar-binding activity. CTLs contain an evolutionarily conserved C-type lectin domain (CTLD) that binds microbial carbohydrates in a calcium-dependent manner, thereby playing a key role in both microbial pathogenesis and innate immune responses. Aedes albopictus is an important vector for transmitting dengue virus (DENV) worldwide. Currently, the molecular characteristics and functions of CTLs in Ae. albopictus are largely unknown. METHODS: Transcripts encoding CTL proteins in the Ae. albopictus genome assembly were analyzed via sequence blast. Phylogenetic analysis and molecular characterization were performed to identify the functional domains of the CTLs. Quantitative analysis was performed to determine the gene expression features of CTLs during mosquito development and in different tissues of female adults after blood feeding. In addition, the functional role of CTLs in response to DENV infection was investigated in Ae. albopictus mosquito cells. RESULTS: We identified 39 transcripts encoding CTL proteins in the Ae. albopictus transcriptome. Aedes albopictus CTLs are classified into three groups based on the number of CTLDs and the domain architecture. These included 29 CTL-Ss (single-CTLDs), 1 immulectins (dual-CTLD) and 9 CTL-Xs (CTLDs with other domains). Phylogenetic analysis and structural modeling indicated that CTLs in Ae. albopictus are highly conserved with the homologous CTLs in Aedes aegypti. The expression profile assay revealed differential expression patterns of CTLs in both developmental stages and in adult female tissues. Knockdown and overexpression of three CTLs (CTL-S12, S17 and S19) confirmed that they can promote dengue virus infection in Ae. albopictus cells. CONCLUSIONS: The CTL genes in Ae. albopictus mosquito and other mosquito species are evolutionarily conserved and exhibit different developmental and tissue expression features. The functional assay indicated that three CTLs in Ae. albopictus mosquitoes are involved in promoting dengue virus infection. Our study revealed that CTLs play important roles in both the physiological processes and viral infection in mosquito vectors.

Abiotic factors shape mosquito microbiomes that enhance host development.

Metazoans rely on interactions with microorganisms through multiple life stages. For example, developmental trajectories of mosquitoes can vary depending on the microorganisms available during their aquatic larval phase. However, the role that the local environment plays in shaping such host-microbe dynamics and the consequences for the host organism remain inadequately understood. Here, we examine the influence of abiotic factors, locally available bacteria, and their interactions on the development and associated microbiota of the mosquito Aedes albopictus. Our findings reveal that leaf detritus infused into the larval habitat water, sourced from native Hawaiian tree "ohi'a lehua Metrosideros polymorpha, invasive strawberry guava Psidium cattleyanum, or a pure water control, displayed a more substantial influence than either temperature variations or simulated microbial dispersal regimes on bacterial community composition in adult mosquitoes. However, specific bacteria exhibited divergent patterns within mosquitoes across detrital infusions which did not align with their abundance in the larval habitat. Specifically, we observed a higher relative abundance of a Chryseobacterium sp. strain in mosquitoes from the strawberry guava infusion than the pure water control, whereas the opposite trend was observed for a Pseudomonas sp. strain. In a follow-up experiment, we manipulated the presence of these two bacterial strains and found larval developmental success was enhanced by including the Chryseobacterium sp. strain in the strawberry guava infusion and the Pseudomonas sp. strain in the pure water control. Collectively, these data suggest that interactions between abiotic factors and microbes of the larval environment can help shape mosquito populations" success.

Widespread geographic distribution of Aedes aegypti (Diptera: Culicidae) kdr variants in Panama.

We searched for evidence of knockdown resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes aegypti (Linnaeus) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) mosquitoes from Panama. Conventional PCR was performed on 469 Ae. aegypti and 349 Ae. albopictus. We did not discover kdr mutations in Ae. albopictus, but 2 nonsynonymous kdr mutations, V1016I (found in 101 mosquitoes) and F1534C (found in 29 of the mosquitoes with the V1016I), were detected in Ae. aegypti. These kdr mutations were present in all specimens that were successfully sequenced for both IIS5-S6 and IIIS6 regions, which included samples collected from 8 of the 10 provinces of Panama. No other kdr mutations were found in Ae. aegypti, including V1016G, which has already been reported in Panama. Findings suggest that the V1016I-F1534C variant is prevalent in Panama, which might be related to the introduction and passive movement of mosquitoes as part of the used-tire trade. However, we cannot rule out the possibility that selection on de novo replacement of kdr mutations also partially explains the widespread distribution pattern of these mutations. These 2 ecological and evolutionary processes are not mutually exclusive, though, as they can occur in tandem. Research in Panama needs to calculate the genotypic and allelic frequencies of kdr alleles in local Ae. aegypti populations and to test whether some combinations confer phenotypic resistance or not. Finally, future studies will have to track the introduction and spreading of new kdr mutations in both Aedes species.

Mesocosm Studies Suggest Climate Change May Release Aedes aegypti (Diptera: Culicidae) Larvae from Cold Inhibition and Enable Year-Round Development in a Desert City.

AbstractGlobal warming trends, human-assisted transport, and urbanization have allowed poleward expansion of many tropical vector species, but the specific mechanisms responsible for thermal mediation of range changes and ecological success of invaders remain poorly understood. Aedes aegypti (Diptera: Culicidae) is a tropical mosquito currently expanding into many higher-latitude regions, including the urban desert region of Maricopa County, Arizona. Here, adult populations virtually disappear in winter and spring and then increase exponentially through summer and fall, indicating that winter conditions remain a barrier to the development of some life stages of A. aegypti. To determine whether cold limits the winter development of A. aegypti larvae in Maricopa County, we surveyed for larval abundance and tested their capacity to develop in ambient and warmed conditions. Aedes aegypti larvae were not observed in artificial aquatic habitats in winter and spring but were abundant in summer and fall, suggesting winter suppression of adults, larvae, or both. Water temperatures in winter months fluctuated strongly; larvae were usually cold paralyzed at night but active during the day. Despite daytime temperatures that allowed activity and achieving similar degree-days as warmed mesocosms, larvae reared under ambient winter conditions were unable to develop to adulthood, perhaps due to repetitive cold damage. However, warming average temperature by 1.7°C allowed many larvae to successfully develop to adults. Because daytime highs in winter will often allow adult flight, it is likely that relatively minor additional winter warming may allow A. aegypti populations to develop and reproduce year-round in Maricopa County.