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Background: The decision to imbibe a blood meal is predominantly dependent on the sensitivity and specificity of haematophagous arthropods to blood-derived adenine nucleotides, in particular adenosine triphosphate (ATP). Despite previous efforts to identify and characterise the specificity and sensitivity to ATP and other adenine nucleotides, as well as the role of other blood-derived phagostimulants across the Culicidae, comparisons across species remain difficult. Methods: The feeding response of the yellow fever mosquito Aedes aegypti and the African malaria vector Anopheles gambiae to adenine nucleotides in the presence of a carbonate buffer was assessed using a membrane feeding assay. The proportion of mosquitoes engorged and the volume imbibed by all mosquitoes was scored visually and spectrophotometrically. In addition, the proportion of prediuresing An. gambiae, as well as the volume engorged and prediuresed, was examined. Results: Aedes aegypti was more sensitive to adenine nucleotides than An. gambiae, but both species maintained specificity to these phagostimulants, demonstrating a dose-dependent bimodal feeding pattern, thereby expanding our understanding of the all-or-none blood-feeding hypothesis. Feeding on the bicarbonate buffer by An. gambiae—but not that of Ae. aegypti—demonstrated a species-specific variation in how blood phagostimulants are encoded. Adenine nucleotides, with and without bovine serum albumin, were observed to dose-dependently regulate the proportion of An. gambiae prediuresing and the volumes prediuresed but not volumes engorged. Conclusions: Taken together, the results of this study expand our understanding of how mosquitoes differentially assess and respond to blood meal constituents, and provide a basis for further physiological and molecular studies. [ABSTRACT FROM AUTHOR]

Wolbachia pipientis (= Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of transient somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB had more modest effects. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit. Author summary: In recent years, wild populations of Aedes aegypti mosquitoes in over a dozen countries have been deliberately infected with Wolbachia pipientis ("Wolbachia"); an intracellular bacterium that, in some circumstances, helps to curb the spread of mosquito-brone pathogens including dengue virus. But how does Wolbachia affect the ability of mosquitoes to become infected with and spread the many different viruses they encounter in nature? Here, we use transient somatic infections in Aedes aegypti to characterize the effects of Wolbachia on three different alphaviruses that cause illness in humans: Sindbis virus, O'nyong-nyong virus, and Mayaro virus. We find that transient Wolbachia infections have variable effects on these different pathogens, ranging from significant suppression of Mayaro virus to significant enhancement of Sindbis virus. Our research has important implications for the design of vector control strategies, and suggests further research is needed to understand how Wolbachia shapes the replication and transmission of diverse viruses in mosquitoes. [ABSTRACT FROM AUTHOR]

BACKGROUND: Controlling the spread of arboviral diseases remains a considerable challenge due to the rapid development of insecticide resistance in Aedes mosquitoes. This study evaluated the effects of boric acid‐containing toxic sugar bait (TSB) on field populations of resistant Aedes aegypti mosquitoes. In addition, this study examined the flight activity and wing beat frequency and amplitude of males and the flight activity, fecundity, and insemination of females after pairing with males exposed to TSB. The population dynamics of Aedes mosquitoes under imbalanced sex ratios were examined to simulate realistic field conditions for male suppression under the effect of TSB. RESULTS: The mortality of male mosquitoes was consistently high within 24 h after exposure. By contrast, the mortality of female mosquitoes was inconsistent, with over 70% mortality observed at 168 h. The flight activity and wing beat amplitude of treated males were significantly lower than those of controls, but no significant difference in wing beat frequency was detected. The fecundity and insemination of treated female mosquitoes were lower than those of controls. A simulation study indicated that considerably low male population densities led to mating failures, triggering a mate‐finding Allee effect and resulting in persistently low population levels. CONCLUSION: Boric acid‐containing TSB could effectively complement current chemical intervention approaches to control resistant mosquito populations. TSB is effective in reducing field male populations and impairing male flight activity and female‐seeking behavior, resulting in decreased fecundity and insemination. Male suppression due to TSB potentially results in a small mosquito population. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Orthoflaviviruses are arthropod-borne viruses that are transmitted by mosquitoes or ticks and cause a range of significant human diseases. Among the most important tick-borne orthoflaviviruses (TBFVs) is tick-borne encephalitis virus (TBEV), which is endemic in Eurasia, and Powassan virus, which is endemic in Asia and North America. There is a significant controversy regarding species assignment in the tick-borne encephalitis virus complex due to the complex phylogenetic, serological, ecological, and pathogenetic properties of viruses. Comparing the rate of non-synonymous to synonymous substitutions (dN/dS) over the course of tick-borne orthoflavivirus diversification suggests that there is a very strong stabilizing selection (Nei-Gojobori dN/dS < 0.1) among tick-borne orthoflaviviruses that differ by less than 13.5% amino acid/21.4% nucleotide sequences, and discretely more rapid accumulation of non-synonymous substitutions (dN/dS > 0.13) among more divergent viruses that belong to distinct species. This pattern was similarly observed in genome regions encoding structural (E) and non-structural (NS3) proteins. Below this distance threshold, viruses appear fit and strongly tied to their ecological niche, whereas above the threshold, a greater degree of adaptation appears necessary. This species criterion suggests that all subtypes of TBEV, all related ovine/caprine encephalomyelitis viruses, and Omsk hemorrhagic fever virus (OHFV) together correspond to a single species. Within this species, viruses make up 11 subtypes that are reliably segregated by a 10% nucleotide distance cut-off suggested earlier for TBEV. The same 10% subtype cut-off suggests that Powassan virus includes two subtypes, Powassan and Deer Tick virus. [ABSTRACT FROM AUTHOR]

Background: Surveillance of the host–anopheline mosquitoes' interaction is important for assessing malaria transmission risk and guiding vector control. We assume that changes in malaria vector species' feeding habits, as well as the surrounding environment, have a substantial impact on varied malaria transmission. In this study, we determined the vertebrate host feeding patterns of anopheline mosquitoes to characterize entomologic risk factors for malaria in Jabi Tehnan, Northwestern Ethiopia. Methods: Blood-fed anophelines surveyed during malaria surveillance in Jabi Tehnan district of northwestern Ethiopia were utilized in this study. They were collected using Centers for Disease Control and Prevention (CDC) light traps deployed in selected households per village, placed indoors and outdoors, spanning three agroecological settings (dry mountain, plateau, and semiarid highlands) between June 2020 and May 2021. The engorged mosquitoes were analyzed for host blood meal sources and Plasmodium infection via polymerase chain reaction (PCR) and/or sequencing. Infection rates and bovine and human blood indices were calculated and compared for abundant species; between indoors and outdoors and between agroecology using a chi-squared test for equality of proportion in R package at a significant level of p ≤ 0.05. Results: A total of 246 mosquitoes were successfully typed (indoor, 121; outdoor, 125), with greater relative abundance indoors in mountain and plateau highlands, and outdoors in semiarid areas. Despite ecological differences in blood-fed capture rates, cattle served as the most utilized blood meal source by 11 anopheline species with an overall bovine blood index (BBI) of 74.4%. This trend was dictated by Anopheles gambiae s.l. (198/246; BBI = 73.7%), which exhibited the most plastic feeding habits that included humans (human blood index = 15.7%) and other livestock and rodents. A total of five anopheline species (An. gambiae s.l., An. funestus s.l., An. coustani s.l., An. pretoriensis, and An. pharoensis) fed on humans, of which the first three were found infected with Plasmodium parasites. Most of the infected specimens were An. arabiensis (5.6%, 11/198) and had recently fed mainly on cattle (72.7%, 8/11); one each of infected An. funestus s.l. and An. coustani s.l. had fed on humans and cattle, respectively. Conclusions: The results demonstrate communal feeding on cattle by anophelines including primary and secondary malaria vectors. This study also indicates the importance of cattle-targeted interventions for sustainable control of malaria vectors in the study areas. [ABSTRACT FROM AUTHOR]

The RNA genome of orthoflaviviruses encodes a methyltransferase within the non-structural protein NS5, which is involved in 2'-O-methylation of the 5'-terminal nucleotide of the viral genome resulting in a cap1 structure. While a 2'-O-unmethylated cap0 structure is recognized in vertebrates by the RNA sensor RIG-I, the cap1 structure allows orthoflaviviruses to evade the vertebrate innate immune system. Here, we analyzed whether the cap0 structure is also recognized in mosquitoes. Replication analyses of 2'-O-methyltransferase deficient yellow fever virus mutants (YFV NS5-E218A) of the vaccine 17D and the wild-type Asibi strain in mosquito cells revealed a distinct downregulation of the cap0 viruses. Interestingly, the level of inhibition differed for various mosquito cells. The most striking difference was found in Aedes albopictus-derived C6/36 cells with YFV-17D cap0 replication being completely blocked. Replication of YFV-Asibi cap0 was also suppressed in mosquito cells but to a lower extent. Analyses using chimeras between YFV-17D and YFV-Asibi suggest that a synergistic effect of several mutations across the viral genome accompanied by a faster initial growth rate of YFV-Asibi cap1 correlates with the lower level of YFV-Asibi cap0 attenuation. Viral growth analyses in Dicer-2 knockout cells demonstrated that Dicer-2 is entirely dispensable for attenuating the YFV cap0 viruses. Translation of a replication-incompetent cap0 reporter YFV-17D genome was reduced in mosquito cells, indicating a cap0 sensing translation regulation mechanism. Further, oral infection of Aedes aegypti mosquitoes resulted in lower infection rates for YFV-Asibi cap0. The latter is related to lower viral loads found in the midguts, which largely diminished dissemination to secondary tissues. After intrathoracic infection, YFV-Asibi cap0 replicated slower and to decreased amounts in secondary tissues compared to YFV-Asibi cap1. These results suggest the existence of an ubiquitously expressed innate antiviral protein recognizing 5'-terminal RNA cap-modifications in mosquitoes, both in the midgut as well as in secondary tissues. Author summary: Arthropod-borne viruses infect both mosquito and vertebrate hosts. One factor influencing the level of viral replication in the host is the ability of the virus to evade or counteract the host's immune system. In vertebrates, it was shown that orthoflaviviruses circumvent the innate immune system by capping their viral genome with a cap1 structure characterized by 2'-O-methylation of the 5'-terminal nucleotide. In contrast, viral genomes lacking 2'-O-methylation (cap0) are recognized as foreign and activate the vertebrate innate immune system. Here, we used yellow fever virus (YFV) strain 17D and Asibi mutants lacking 2'-O-methylation, as well as a cap0 reporter YFV genome, to analyze cap0 recognition in mosquito cells and living mosquitoes. The replication of the YFV cap0 mutants was profoundly impaired in different mosquito cells and cap0 YFV genomes showed reduced translation levels. Based on viral infection experiments in Aedes aegypti mosquitoes, we further demonstrate that these vector mosquitoes recognize the cap0 structure and that a potential recognizing effector protein is localized in the midgut as well as in secondary tissues. Our findings provide the basis for future studies investigating the recognition of foreign RNAs in mosquitoes and should contribute to establish methods to prevent transmission of arboviruses. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the chemical composition of essential oils from Myroxylon peruiferum and Eugenia gracillima and determine the larvicidal potential of the oils against Aedes aegypti. Essential oils from the leaves and young stems of M. peruiferum (EOLMP and EOAMP) and the leaves of E. gracillima (EOLEG) were obtained through hydrodistillation. The major constituents of EOLMP were α-pinene (18.0%) and δ-cadinene (14.2%); the major constituents of EOAMP were spathulenol (22.9%) and β-elemene (12.4%) and the major constituents of EOLEG were δ-elemene (40.4%) and germacrene D (25.2%). The two oils extracted from M. peruiferum were very active against A. aegypti larvae and had similar lethal concentrations (LC50 = 0.7 mg/mL). The oil extracted from E. gracillima had an LC50 of 1.51 mg/mL. The essential oils from M. peruiferum and E. gracillima constitute promising sources for the formulation of novel insecticides for the control of the mosquito Aedes aegypti. [ABSTRACT FROM AUTHOR]

We report a convergent synthesis of heteroaryl/aryl-annulated pyridine and quinoline derivatives by a metal-free Povarov reaction. para -Toluene sulfonic acid was used as the catalyst in this reaction, which produced the products in good yields from the corresponding aromatic amines and ethyl vinyl ether. A pyridocoumarin and a pyridopyrimidine product were evaluated for their mosquito larvicidal activity against the third instar larvae of the dengue vector mosquito Aedes aegypti. Examination of morphological changes in the larvae showed damage to the target body part after treatment with both the pyridocoumarin and pyridopyrimidine products at the LC50 concentrations. [ABSTRACT FROM AUTHOR]

Mosquitoes are significant vectors of deadly diseases such as malaria and dengue, contributing to approximately 300,000 deaths annually, according to the World Health Organization (WHO). Traditional chemical repellents, while effective, often cause severe allergic reactions and contribute to mosquito resistance. This study explores the formulation of GAP-F essential oil-loaded microspheres as a natural and eco-friendly alternative for mosquito repellency. Essential oils have demonstrated insect-repellent properties; however, their volatility limits their effectiveness. Microencapsulation is employed to protect these oils from degradation and facilitate sustained release, enhancing their efficacy against mosquitoes. GAP-F essential oil was sourced from the CSIR-CIMAP laboratory through distillation. Laboratory tests were conducted to evaluate its effectiveness against Aedes aegypti, Anopheles stephensi and wild mosquitoes. At a concentration of 10%, GAP-F essential oil exhibited repellency rates of 70%, 85% and 75% against these species during a one-hour bioassay. Given the volatile nature of essential oils, the formulation of microspheres serves as a protective carrier that enhances stability and prolongs the duration of protection. The essential oil-loaded microspheres demonstrated satisfactory results in terms of repellency, showing effectiveness rates of 56.66%, 51.66% and 55% against Aedes aegypti, Anopheles stephensi and wild mosquitoes, respectively, during the same duration. Mosquitoes aged between three to five days were used for the bioassay. This research highlights the potential of microencapsulated natural repellents as a viable alternative to chemical products in mosquito control strategies, providing an effective solution that minimizes health risks while addressing the growing concern of insecticide resistance in mosquito populations. [ABSTRACT FROM AUTHOR]

Mosquitoes serve as vectors for many arthropod-borne viruses (arboviruses) that are responsible for millions of human infections and thousands of deaths each year. Among these arboviruses, O'nyong-nyong virus (ONNV) is an African alphavirus mainly transmitted by Anopheles mosquitoes. ONNV can be detected through serological or molecular tests, the first showing cross-reactivity to co-circulating alphaviruses and requiring technically demanding confirmation, while the latter, usually based on real-time PCR, are costly and demand specific equipment. Isothermal amplification approaches, such as Loop-Mediated Isothermal Amplification (LAMP), should therefore provide a cost-effective, sensitive, and specific alternative for virus detection, suitable for the resource-limited regions where ONNV circulates up to the present time. Here, we describe the development and optimization of a rapid and highly sensitive (10 pfu/reaction) RT-LAMP assay for ONNV detection. Additionally, we demonstrate that it is possible to bypass the RNA extraction step, reducing sample handling time and costs. The final RT-LAMPONNV is a promising field detection tool for ONNV, enabling a better understanding of its impact and serving as a point-of-care diagnostic method. [ABSTRACT FROM AUTHOR]

The global distribution of Aedes aegypti mosquitoes, particularly in tropical regions, poses a significant public health risk due to their apparent ability to transmit arboviruses such as West Nile virus (WNV). This study aimed to evaluate the vector competence of Ae. aegypti from São Tomé and Príncipe (STP) for the transmission of the WNV PT6.39 strain, considering its potential role as a bridge vector in a region where Culex quinquefasciatus would be the main vector. Aedes aegypti mosquitoes were collected, reared, and experimentally infected with WNV, with viral dissemination and transmission potential assessed 7, 14, and 21 days post infection (dpi). The results showed an increasing trend in infection rates, from 5% at 7 dpi to 35% at 21 dpi, with corresponding dissemination rates of 0%, 100%, and 43%. The transmission rates also increased from 0% at 7 dpi to 67% at 21 dpi, with a maximum transmission efficiency of 10% observed at the final time point. Although Ae. aegypti from STP demonstrated the potential to transmit WNV, the overall transmission efficiency remained relatively low. These findings provide necessary insights into the vector competence of Ae. aegypti in this region, highlighting the importance of continued monitoring and targeted vector control measures to mitigate the risk of potential WNV outbreaks. [ABSTRACT FROM AUTHOR]

Wolbachia are the most spread bacterial endosymbionts in the world. These bacteria can manipulate host reproduction or block virus transmission in mosquitoes. For this reason, Wolbachia-based strategies for vector control are seriously considered or have already been applied in several countries around the world. In South America, Wolbachia have been studied in human pathogen vectors such as sand flies and mosquitoes. In French Guiana, the diversity and distribution of Wolbachia are not well known in mosquitoes. In this study, we screened for Wolbachia natural infection in mosquitoes in French Guiana by using 16S rRNA, Wolbachia surface protein (WSP), and multi-locus sequence typing (MLST) molecular assays. A total of 29 out of 44 (65.9%) mosquito species were positive for natural Wolbachia infection according to the PCR results, and two Wolbachia strains co-infected three specimens of Mansonia titillans. Then, we analyzed the phylogenetic relationships among the Wolbachia detected. All of the tested specimens of Aedes aegypti, the major dengue vector of French Guiana, were negative. These results regarding Wolbachia strain, distribution, and prevalence in mosquitoes from French Guiana highlight Wolbachia–mosquito associations and pave the way for a future Wolbachia-based strategy for vector control in this Amazonian territory. [ABSTRACT FROM AUTHOR]

Issue Addressed Methods Results Conclusions So What? An important part of preventing and managing Aedes‐borne mosquito disease outbreak risk is engaging the community. Research shows that high‐income countries tend to use top‐down measures for Aedes mosquito management, favouring educational approaches to engage the community over participatory approaches that actively involve and empower the community in addressing disease risk. Little is known about the reasons behind the use of these approaches and how they could be strengthened. This research explores the community engagement approaches used in Aedes mosquito management in Cairns, Queensland, Australia and the factors influencing the choice of these approaches.A case study design was used, drawing on two qualitative methods—key informant, semi‐structured interviews (n = 25), and a document review (n = 20). Thematic analysis was used to identify, analyse and attribute meaning from the data.Various approaches were used to engage the community, including direct interaction through door‐to‐door inspections, broad outreach via mass media campaigns, and community participation in a novel mosquito replacement strategy. Factors influencing the choice of these approaches included government legislative responsibilities, research‐related ethical obligations, work norms within local government and public health units, the perceived importance of gaining community trust, constraints on workforce capacity, time and funding.There were multiple factors influencing the community engagement approaches used in this study. Resource constraints, institutional norms and prevailing attitudes and beliefs were identified as hindering the use of more empowering approaches to engaging the community. These barriers should be considered and addressed in the planning of Aedes mosquito management to better support community engagement in this setting.Community engagement is an important aspect of managing the Aedes mosquito disease threat. With the global increase in Aedes mosquito‐borne disease risk, these findings can help other at‐risk settings understand potential organisational impediments to engaging the community. This is particularly important when advocating for the inclusion of bottom‐up approaches in policy, and to ensure sufficient resources are allocated to strengthen community engagement in Aedes mosquito management. [ABSTRACT FROM AUTHOR]

The main vectors of Zika virus (ZIKV) and dengue virus (DENV) are Aedes aegypti and Ae. albopictus, with Ae. aegypti being more competent. However, the underlying mechanisms remain unclear. Here, we find Ae. albopictus shows comparable vector competence to ZIKV/DENV with Ae. aegypti by blood-feeding after antibiotic treatment or intrathoracic injection. This suggests that midgut microbiota can influence vector competence. Enterobacter hormaechei_B17 (Eh_B17) is isolated from field-collected Ae. albopictus and conferred resistance to ZIKV/DENV infection in Ae. aegypti after gut-transplantation. Sphingosine, a metabolite secreted by Eh_B17, effectively suppresses ZIKV infection in both Ae. aegypti and cell cultures by blocking viral entry during the fusion step, with an IC50 of approximately 10 μM. A field survey reveals that Eh_B17 preferentially colonizes Ae. albopictus compared to Ae. aegypti. And field Ae. albopictus positive for Eh_B17 are more resistant to ZIKV infection. These findings underscore the potential of gut symbiotic bacteria, such as Eh_B17, to modulate the arbovirus vector competence of Aedes mosquitoes. As a natural antiviral agent, Eh_B17 holds promise as a potential candidate for blocking ZIKV/DENV transmission. Here the authors show that arbovirus vector competence of Aedes mosquitoes is modulated by the midgut microbiota and identify the metabolite sphingosine from the gut symbiotic bacterium Enterobacter hormaechei_B17 (Eh_B17) as a factor mediating resistance to ZIKV and DENV infection. [ABSTRACT FROM AUTHOR]

Mosquitoes transmit viruses that affect public health in tropical regions. Identifying factors that influence their fitness can help control mosquito-borne diseases. We investigated the impacts of predation risk effects, from a fish predator, and food availability on the oviposition behavior of Aedes aegypti. We hypothesize that predation risk and food availability interactively affect Ae. aegypti oviposition. Gravid Ae. aegypti females were offered oviposition sites with all possible combinations of predation risk (with vs. without predator cues) and food availability (low vs. high). We found a significant interaction between predation risk and food availability. The magnitude of oviposition and the probability of egg laying in high food availability treatments exceeded the values of low food availability treatments, but only in treatments without predation risk. Predation risk critically modulated the effects of food availability on the oviposition of Ae. aegypti. This study demonstrates the interplay between predation risk and food availability in controlling mosquito-borne diseases. [ABSTRACT FROM AUTHOR]

Mosquito-borne diseases are a major global health threat. Traditional morphological or molecular methods for identifying mosquito species often require specialized expertise or expensive laboratory equipment. The use of convolutional neural networks (CNNs) to identify mosquito species based on images may offer a promising alternative, but their practical implementation often remains limited. This study explores the applicability of CNNs in classifying mosquito species. It compares the efficacy of body and wing depictions across three image collection methods: a smartphone, macro-lens attached to a smartphone and a professional stereomicroscope. The study included 796 specimens of four morphologically similar Aedes species, Aedes aegypti, Ae. albopictus, Ae. koreicus and Ae. japonicus japonicus. The findings of this study indicate that CNN models demonstrate superior performance in wing-based classification 87.6% (95% CI: 84.2–91.0) compared to body-based classification 78.9% (95% CI: 77.7–80.0). Nevertheless, there are notable limitations of CNNs as they perform reliably across multiple devices only when trained specifically on those devices, resulting in an average decline of mean accuracy by 14%, even with extensive image augmentation. Additionally, we also estimate the required training data volume for effective classification, noting a reduced requirement for wing-based classification compared to body-based methods. Our study underscores the viability of both body and wing classification methods for mosquito species identification while emphasizing the need to address practical constraints in developing accessible classification systems. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Insect-specific viruses (ISVs) include viruses that are restricted to the infection of mosquitoes and are spread mostly through transovarial transmission. Despite using a distinct mode of transmission, ISVs are often phylogenetically related to arthropod-borne viruses (arboviruses) that are responsible for human diseases and able to infect both mosquitoes and vertebrates. ISVs can also induce a phenomenon called "superinfection exclusion", whereby a primary ISV infection in an insect inhibits subsequent viral infections of the insect. This has sparked interest in the use of ISVs for the control of pathogenic arboviruses transmitted by mosquitoes. In particular, insect-specific flaviviruses (ISFs) have been shown to inhibit infection of vertebrate-infecting flaviviruses (VIFs) both in vitro and in vivo. This has shown potential as a new and ecologically friendly biological approach to the control of arboviral disease. For this intervention to have lasting impacts for biological control, it is imperative that ISFs are maintained in mosquito populations with high rates of vertical transmission. Therefore, these strategies will need to optimise vertical transmission of ISFs in order to establish persistently infected mosquito lines for sustainable arbovirus control. This review compares recent observations of vertical transmission of arboviral and insect-specific flaviviruses and potential determinants of transovarial transmission rates to understand how the vertical transmission of ISFs may be optimised for effective arboviral control. [ABSTRACT FROM AUTHOR]

Oral infection of mosquitoes by arboviruses often results in a large degree of variation in the amount of infectious virus between individual mosquitoes, even when the mosquitoes are from inbred laboratory strains. This variability in arbovirus load has been shown to affect virus transmissibility. Previously, our group described population genetic and specific infectivity differences between the virus populations found in high and low titer Aedes aegypti mosquitoes that had been orally infected with Sindbis virus (SINV). In this study, we sought to investigate whether there were also differences in transcriptomic response between these high and low titer mosquitoes. Results from the transcriptomic data analysis showed that more genes involved in antiviral activity, endopeptidase activity, and methyltransferase activity were upregulated in low titer mosquitoes than in high titer mosquitoes, relative to blood-fed controls. Meanwhile, genes involved in ion transport, energy metabolism, acetylation, glycosylation, lipid metabolism, and transport tended to be upregulated in high titer mosquitoes more than in low titer mosquitoes, relative to blood-fed mosquitoes. Overall, genes involved in antiviral activities tended to be upregulated in low titer mosquitoes while genes involved in proviral activities were mostly upregulated in high titer mosquitoes. This study has identified a number of candidate mosquito genes that are putatively associated with SINV titer variability after oral infection of Ae. aegypti, and these can now be investigated in order to ascertain their roles in virus replication and their contributions to determining vector competence. [ABSTRACT FROM AUTHOR]

Background: This study conducted in the departments of Oueme and Plateau aims to assess the presence of the dengue virus and its different serotypes in Aedes aegypti and Aedes albopictus, as well as the epidemic risk incurred by the populations. Methods: Collections of adult mosquitoes using human landing catches (HLC) were carried out in six communes, three (Porto-Novo, Adjarra, and Avrankou) in the Oueme department and the rest (Ifangni, Kétou, and Pobè) in the Plateau department. Pools of ten Aedes mosquitoes were formed, and stored at −80°C in RNA later. RT-PCR was used to detect dengue virus, and conventional PCR for the different serotypes. Inspection of water containers and collection of Aedes larvae was performed inside and around each house to calculate the stegomyan indices. Results: In the six communes, the dengue virus was present both in Ae. aegypti and Ae. albopictus. Combined data of the two Aedes species at the communes level revealed infection rates ranging from 80.00% (95% CI: 61.43–92.29) to 96.67% (95% CI: 82.78–99.92). In all the communes, the values of stegomyan indices reached the WHO threshold, which indicates the existence of the risk of an arbovirus epidemic. In addition, the infection rates were similar for Ae. aegypti [88.19% (95% CI: 81.27–93.24)] and Ae. albopictus [86.79% (95% CI: 74.66–94.52)]. The three virus serotypes detected in the pools of Aedes were DENV-1, DENV-3, and DENV-4, with a high prevalence for the first two. Conclusion: This study revealed that three serotypes (DENV-1, DENV-3, and DENV-4) of dengue virus circulate in Ae. aegypti and Ae. albopictus in the departments of Oueme and Plateau. Moreover, the risk of transmission of arboviruses was globally high and variable from commune to commune. This information is essential for informed decision-making in the preventive control of the disease. [ABSTRACT FROM AUTHOR]

Insects geographically separated into island and mainland populations often exhibit phenotypic variations, a phenomenon known as insular conditions. These conditions can lead to rapid evolutionary changes that affect the morphological characteristics of mosquito vectors. Nevertheless, studies that specifically examine phenotype differences between island and mainland mosquito populations have been limited. In this study, wing variation in size and shape was investigated using the geometric morphometric (GM) technique in two dominant mosquito vectors, Aedes albopictus and Armigeres subalbatus, in the Ranong and Trat archipelagos of Thailand. Significant differences in average wing centroid size (CS) were found in 6 out of 15 population pairs for Ae. albopictus (p < 0.05) and in 5 population pairs for Ar. subalbatus (p < 0.05). After removing the allometric effect, canonical variate analyses (CVA) based on wing shape analysis revealed overlap across all populations for both Ae. albopictus and Ar. subalbatus. However, the statistical analysis indicated that Ar. subalbatus exhibited wing shape differences across all populations (p < 0.05), and most Ae. albopictus populations also displayed distinct wing shapes (p < 0.05), except for the populations from Chang Island and the mainland of Ranong, which showed no significant differences (p > 0.05). These findings enhance our understanding of mosquito adaptability in island regions and provide valuable data for the surveillance and monitoring of vector evolution. [ABSTRACT FROM AUTHOR]

In an age of emerging zoonoses, it is important to understand the intricate system of vectors and reservoirs, or hosts, and their relation to humans. West Nile Virus (WNV) has been detected in a myriad of nonhuman hosts. Transmission of the virus to humans is reliant on amplified seroprevalence within the host, which occurs primarily in birds. However, recent studies have found that other animal groups, including crocodilians, can obtain seroprevalence amplification to levels that make them competent hosts able to transmit WNV to mosquitoes, which can then transmit to humans. Climate change could exacerbate this transmission risk by shifting the distributions of mosquito vectors towards novel geographic ranges. Here, we use maximum entropy models to map the current and future distributions of three mosquito vector species and four crocodilian species in North America to determine the emerging risk of WNV outbreaks associated with changing climates and WNV associated with crocodilians in North America. From our models, we determined that one mosquito species in particular, Culex quinquefasciatus, will increase its distribution across the ranges of all crocodilian species in all tested climate change scenarios. This poses a potential risk to public health for people visiting and living near crocodilian farms and high-density natural crocodilian populations. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Mosquito traps, historically used for surveillance and research, have gained prominence as a tool for mosquito control, amidst concern over the environmental impact and increased resistance to insecticide‐based methods. In this study, we tested the effectiveness of a mass trapping barrier design with two types of traps, Mosquito Magnet (MM) traps and BG‐Protector (BGP) traps. This experiment was conducted in three coastal camping areas in southern France between summer and autumn 2022, where the presence of floodwater mosquito species with anthropophilic preferences like Aedes caspius represents a year‐long nuisance. MM traps were set around the campsite as a barrier to interfere with mosquitoes from entering the campsites, whereas BGP traps were set within the campsites, with the aim of diverting mosquitoes away from humans at peak activity hours. Over 210,000 mosquitoes of 11 species from 4 genera were collected by both trap types across treatment campsites, with no significant differences in mosquito community samplings between BGP and MM traps. Barrier traps effectively targeted Ae. caspius, reducing total mosquito abundance in two of the three study sites by 34% and 55%. This study provides valuable insights into the efficacy and feasibility of using mass trapping barriers as a complementary control strategy for mosquito species in wetlands. [ABSTRACT FROM AUTHOR]

BACKGROUND: The incompatible insect technique (IIT) has been used for Aedes mosquito population suppression to curb the transmission of dengue. However, its wide application is limited owing to the low output of male mosquitoes and the risk of population replacement from the release of fertile Wolbachia‐infected females. This study aims to improve IIT efficiency for broader adoption. RESULTS: We assessed the impact of 10% pyriproxyfen (PPF) sticky powder exposure on Wolbachia (from Culex molestus)‐transinfected Aedes albopictus Guangzhou line (GUA line) (GC) mosquitoes. We found that the exposure caused chronic toxicity in adult mosquitoes without affecting the cytoplasmic incompatibility (CI)‐inducing capability of males. The PPF‐contaminated GC females exhibited significant sterilization and the ability to disseminate lethal doses of PPF to breeding sites. Subsequently, we conducted a field trial combining PPF with IIT aiming to suppress the Ae. albopictus population. This combined approach, termed boosted IIT (BIIT), showed a notable enhancement in population suppression efficiency. The improved efficacy of BIIT was attributed to the dispersion of PPF particles in the field via the released PPF‐contaminated male mosquitoes. During the BIIT field trial, no Wolbachia wPip‐positive Ae. albopictus larvae were detected, indicating the effective elimination of the risk of Wolbachia‐induced population replacement. Additionally, the field trial of BIIT against Ae. albopictus resulted in the suppression of the nontarget mosquito species Culex quinquefasciatus. CONCLUSION: Our results highlight the remarkable efficiency and feasibility of combining IIT with PPF in suppressing mosquito populations, facilitating the widespread implementation of IIT‐based management of mosquito‐borne diseases. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The detection of insecticide resistance in male mosquitoes has been treated with less importance in monitoring insecticide resistance spread in mosquitoes. There are no studies on the susceptibility and synergistic activity of DDT and lambda-cyhalothrin on male Anopheles gambiae and Aedes aegypti in Delta State, Nigeria. Even though studies have extensively reported resistance in female mosquitoes, the susceptibility of male mosquitoes to insecticide classes should be ascertained. In this study, we tested the susceptibility status and synergistic activity of DDT and Lambda-cyhalothrin on An. gambiae and Ae. aegypti in Delta State, Nigeria, in order to ascertain the level of resistance and knockdown. In addition, we modelled the knockdown time using Probit analysis model. WHO bioassay method was used to expose two days old adult mosquitoes to 4% DDT and 0.05% lambda-cyhalothrin. The results showed that An. gambiae mosquitoes exposed to DDT and lambda-cyhalothrin were confirmed resistant (61% and 53% respectively). However, pre-exposing the resistant mosquito population to piperonyl butoxide (4%) showed an increase in mortality to 90% (possible resistance) in DDT and 98% (susceptible) in lambda-cyhalothrin. Ae. aegypti mosquitoes exposed to DDT were susceptible (98%) while those exposed to lambda-cyhalothrin were confirmed resistant (87%) and this increased to complete mortality (100%) in PBO+lambda-cyhalothrin population. Furthermore, the results showed that the knockdown time (KDT50 and KDT95) in An. gambiae exposed to DDT was 39.5–71.2 minutes and 124.5–146.4 minutes respectively, while that of lambda-cyhalothrin was 33.0–81.8 minutes and 64.0–124.4 minutes respectively. In Ae. aegypti, KDT50 and KDT95 was 23.9 and 61.7minutes for DDT exposure whereas it was 5.6–15.3 minutes and 36.1–72.3 minutes for lambda-cyhalothrin exposure. It can be concluded that male An. gambiae mosquitoes exposed to the insecticides were resistant and the causes may be linked to certain resistant genes in the mosquitoes. The chances of transferring resistance are possible in wild species and molecular-based studies on the resistant gene in male mosquitoes as well as the tendencies of transfer are required to establish this focus. [ABSTRACT FROM AUTHOR]

Background: Understanding the relationship between blood-feeding and mating is important in effectively managing the most well-adapted vector insect, Aedes aegypti (Linnaeus). Although extensive studies have investigated the behavioural aspects of Aedes such as blood-feeding, mating, and their relationship, several knowledge gaps still exist. Therefore, the present study was undertaken to determine the possibility of successful mating by unfed, engorged, and partially to fully gravid (up to 5 days after blood-feeding with fully developed eggs) female Ae. aegypti mosquitoes and production of viable eggs. Methods: Mating of sexually mature adult Aedes aegypti was allowed in three different ways. In control 1, the females were allowed to mate before taking blood meal, and in control 2, the females were not at all allowed to mate. In the experiment, the females were separated into six categories, viz. D-0 to D-5. In D-0, the females were allowed to mate immediately after the bloodmeal and, in D-1, the females were allowed to mate on the first day of blood feeding, likewise, the females of D-2, D-3, D-4 and D-5 were allowed to mate on 2nd, 3rd, 4th and 5th day of blood feeding. Ovitrap was uniformly kept on the 4th day of blood feeding for the cages D-0 to D-3 for 1 h and then removed and for the cages D-4, and D-5, the ovitrap was kept on 4th and 5th day of blood feeding for 1h immediately after mating. The total number of eggs and the total number of hatching were counted. In the subsequent days, the entire experiment was replicated two times with different cohorts of mosquitoes, and the mean value of three experiments was used to draw Excel bars with 5% error bars and also for the statistical analysis. Results: It was found that mating just before oviposition was sufficient to produce 1581 eggs (70% compared with control) and fertilize 1369 eggs (85% compared with total eggs laid), which is far higher than the 676 non-hatching (unfertilized) eggs (30%) laid by unmated females. Although mating is not essential for producing eggs, our study shows that even brief exposure to the semen and seminal fluids greatly enhances the oviposition and hatching efficiency, even if the mating occurs just before oviposition. However, those females mating before blood-feeding and those mating after blood-feeding produced 2266 and 2128 eggs, with hatching rates of 96.78% and 95.54%, respectively. Hence, the retention time of seminal fluid in the female seems to influence the number of eggs laid and the number of eggs hatched. Conclusions: In general, mating is possible in Ae. aegypti even minutes before oviposition and is sufficient to produce a greater number of viable eggs. [ABSTRACT FROM AUTHOR]

Beauveria bassiana has potential for Aedes aegypti biological control. However, its efficacy depends on the strain's geographic location, host susceptibility, and virulence. The present study aimed to evaluate the effectiveness of B. bassiana strain BBPTG4 conidia in controlling Ae. aegypti adults and its detection via introns profile on exposed mosquito corpses. Morphologic characteristics among strains were highly similar. Comprehensive testing of these strains demonstrated that BBPT4 exhibited the ideal biological activity for Ae. aegypti control, with a median lethal time (TL50) of 7.5 d compared to ~3 d and ~10 d for BB01 and BB37 strains, respectively. Infected mosquitoes died after GHA and BBPTG4 exposure, and corpses were analyzed for infecting strains detection. Differences among the seven evaluated strains were determined, assessing five different insertion group I intron profiles in BBTG4, BB01, GHA, BB37, and BB02 strains. Mosquitoes infected by BBPTG4 and non-exposed (negative control) intron profiles were obtained. We detected the presence of introns in the BBPTG4 strain, which were not present in non-exposed mosquitoes. In conclusion, B. bassiana strains showed similarities in terms of their cultural and microscopic morphological characteristics and biologicals virulence level, but different intron profiles. BBPTG4 strain-infected Ae. aegypti adult corpses, showing specific amplicons, enabled us to identify B. bassiana at the strain level among infected mosquitoes. However, monitoring and detection of field-infected insects is essential for further verification. [ABSTRACT FROM AUTHOR]

Mosquitoes such as Aedes aegypti must consume a blood meal for the nutrients necessary for egg production. Several transcriptome and proteome changes occur post‐blood meal that likely corresponds with codon usage alterations. Transfer RNA (tRNA) is the adapter molecule that reads messenger RNA codons to add the appropriate amino acid during protein synthesis. Chemical modifications to tRNA enhance codon decoding, improving the accuracy and efficiency of protein synthesis. Here, we examined tRNA modifications and transcripts associated with the blood meal and subsequent periods of vitellogenesis in A. aegypti. More specifically, we assessed tRNA transcript abundance and modification levels in the fat body at critical times post blood‐feeding. Based on a combination of alternative codon usage and identification of particular modifications, we discovered that increased transcription of tyrosine tRNAs is likely critical during the synthesis of egg yolk proteins in the fat body following a blood meal. Altogether, changes in both the abundance and modification of tRNA are essential factors in the process of vitellogenin production after blood‐feeding in mosquitoes. [ABSTRACT FROM AUTHOR]

Recent reports have disclosed the occurrence of Wolbachia in Aedes aegypti. Our study detected Wolbachia infection in Ae. aegypti by screening wild adult mosquitoes using two Wolbachia‐specific molecular markers. Overall, 444 adult Ae. aegypti mosquitoes were collected from April 2022 to October 2022 in Jeddah, Saudi Arabia. Each individual sample was processed and screened for the presence of Wolbachia using selected markers, the Wolbachia‐specific 16S rDNA and the Wolbachia surface protein gene (wsp), under optimized polymerase chain reaction (PCR) conditions, and sequenced. In total, 39 (8.78%) and 48 (10.81%) individual mosquito samples were determined to be infected with Wolbachia using the wsp and 16S rDNA markers, respectively. By utilizing two Wolbachia‐specific molecular markers, our study demonstrated the presence of Wolbachia from individual Ae. aegypti samples. Our results showed a low rate of Wolbachia infection and inferred that the detected strain belongs to supergroup B. [ABSTRACT FROM AUTHOR]

Culex tarsalis Coquillett (Diptera: Culicidae) mosquitoes are capable of vectoring numerous pathogens affecting public and animal health. Unfortunately, the probing behaviors of mosquitoes are poorly understood because they occur in opaque tissues. Electropenetrography (EPG) has the potential to elucidate these behaviors by recording the electrical signals generated during probing. We used an AC–DC EPG with variable input resistors (Ri levels) to construct a waveform library for Cx. tarsalis feeding on human hands. Biological events associated with mosquito probing were used to characterize waveforms at four Ri levels and with two electrical current types. The optimal settings for EPG recordings of Cx. tarsalis probing on human hands was an Ri level of 107 Ohms using an applied signal of 150 millivolts alternating current. Waveforms for Cx. tarsalis included those previously observed and associated with probing behaviors in Aedes aegypti L. (Diptera: Culicidae): waveform families J (surface salivation), K (stylet penetration through the skin), L (types 1 and 2, search for a blood vessel/ingestion site), M (types 1 and 2, ingestion), N (type 1, an unknown behavior which may be a resting and digestion phase), and W (withdrawal). However, we also observed variations in the waveforms not described in Ae. aegypti, which we named types L3, M3, M4, and N2. This investigation enhances our understanding of mosquito probing behaviors. It also provides a new tool for the automated calculation of peak frequency. This work will facilitate future pathogen acquisition and transmission studies and help identify new pest and disease management targets. [ABSTRACT FROM AUTHOR]

Wolbachia symbionts introduced into Aedes mosquitoes provide a highly effective dengue virus transmission control strategy, increasingly utilised in many countries in an attempt to reduce disease burden. Whilst highly effective against dengue and other positive-sense RNA viruses, it remains unclear how effective Wolbachia is against negative-sense RNA viruses. Therefore, the effect of Wolbachia on Bunyamwera virus (BUNV) infection in Aedes aegypti was investigated using wMel and wAlbB, two strains currently used in Wolbachia releases for dengue control, as well as wAu, a strain that typically persists at a high density and is an extremely efficient blocker of positive-sense viruses. Wolbachia was found to reduce BUNV infection in vitro but not in vivo. Instead, BUNV caused significant impacts on density of all three Wolbachia strains following infection of Ae. aegypti mosquitoes. The ability of Wolbachia to successfully persist within the mosquito and block virus transmission is partially dependent on its intracellular density. However, reduction in Wolbachia density was not observed in offspring of infected mothers. This could be due in part to a lack of transovarial transmission of BUNV observed. The results highlight the importance of understanding the complex interactions between multiple arboviruses, mosquitoes and Wolbachia in natural environments, the impact this can have on maintaining protection against diseases, and the necessity for monitoring Wolbachia prevalence at release sites. [ABSTRACT FROM AUTHOR]

Simple Summary: The main mosquito species capable of transmitting arboviruses belong to the genera Aedes spp., Psorophora spp., Anopheles spp., Culex spp., Mansonia spp., Coquillettidia spp., Haemagogus spp., Sabethes spp., Culiseta spp., and Wyeomyia spp. Some neglected mosquito species have the potential to become significant disease vectors due to parameters such as global distribution, rapid adaptation to urban areas, and anthropophilic habits. This review discusses the epidemiological importance and biology of three neglected mosquitoes, Aedes scapularis, Aedes vittatus, and Aedes japonicus japonicus, in the context of vectorial capacity and how urbanization, climate change, and globalization alter disease transmission dynamics and may increase the participation of neglected species in propagating diseases. More than 3550 species of mosquitoes are known worldwide, and only a fraction is involved in the transmission of arboviruses. Mosquitoes in sylvatic and semi-sylvatic habitats may rapidly adapt to urban parks and metropolitan environments, increasing human contact. Many of these mosquitoes have been found naturally infected with arboviruses from the Alphaviridae, Flaviviridae, and Bunyaviridae families, with many being the cause of medically important diseases. However, there is a gap in knowledge about the vector status of newly invasive species and their potential threat to human and domestic animal populations. Due to their rapid distribution, adaptation to urban environments, and anthropophilic habits, some neglected mosquito species may deserve more attention regarding their role as secondary vectors. Taking these factors into account, we focus here on Aedes (Ochlerotatus) scapularis (Rondani), Aedes japonicus japonicus (Theobald), and Aedes (Fredwardsius) vittatus (Bigot) as species that have the potential to become important disease vectors. We further discuss the importance of these neglected mosquitoes and how factors such as urbanization, climate change, and globalization profoundly alter the dynamics of disease transmission and may increase the participation of neglected species in propagating diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: Over the last decade, optical sensors have demonstrated great potential to provide complementary data to monitor insect abundance. This article introduces a field-deployable and non-destructive optical instrument to monitor the abundance of flying insects, called an Entomological Bistatic Optical Sensor System, or eBoss. The study focuses on comparing abundance measurements made by the eBoss and physical traps over an 8-month field campaign. The eBoss made over 302,000 insect observations and evaluated the aerial density (#/m3) of all flying insects as well as specifically male and female mosquitoes with a 1 min resolution, allowing us to monitor both the abundance over the season and daily peak of activity. The study's objectives were to validate the optical sensor's data against physical trap collections, which confirmed the correlation between the two methods. However, the eBoss demonstrated superior temporal resolution (1 min versus approximately 3 days) and statistical power due to its larger sample size. These findings suggest that an eBoss can significantly enhance flying insect monitoring efforts, such as mosquitoes or pollinators, providing valuable insights for vector control strategies, agriculture and public health planning. Optical sensors have shown significant promise in offering additional data to track insect populations. This article presents a comparative study between abundance measurements obtained from a novel near-infrared optical sensor and physical traps. The optical instrument, named an Entomological Bistatic Optical Sensor System, or eBoss, is a non-destructive sensor operating in the near-infrared spectral range and designed to continuously monitor the population of flying insects. The research compares the mosquito aerial density (#/m3) obtained through the eBoss with trap counts from eight physical traps during an eight-month field study. The eBoss recorded over 302,000 insect sightings and assessed the aerial density of all airborne insects as well as male and female mosquitoes specifically with a resolution of one minute. This capability allows for monitoring population trends throughout the season as well as daily activity peaks. The results affirmed the correlation between the two methods. While optical instruments do not match traps in terms of taxonomic accuracy, the eBoss offered greater temporal resolution (one minute versus roughly three days) and statistical significance owing to its much larger sample size. These outcomes further indicate that entomological optical sensors can provide valuable complementary data to more common methods to monitor flying insect populations, such as mosquitoes or pollinators. [ABSTRACT FROM AUTHOR]

Multiple Wolbachia strains can block pathogen infection, replication and/or transmission in Aedes aegypti mosquitoes under both laboratory and field conditions. However, Wolbachia effects on pathogens can be highly variable across systems and the factors governing this variability are not well understood. It is increasingly clear that the mosquito host is not a passive player in which Wolbachia governs pathogen transmission phenotypes; rather, the genetics of the host can significantly modulate Wolbachia‐mediated pathogen blocking. Specifically, previous work linked variation in Wolbachia pathogen blocking to polymorphisms in the mosquito alpha‐mannosidase‐2 (αMan2) gene. Here we use CRISPR‐Cas9 mutagenesis to functionally test this association. We developed αMan2 knockouts and examined effects on both Wolbachia and virus levels, using dengue virus (DENV; Flaviviridae) and Mayaro virus (MAYV; Togaviridae). Wolbachia titres were significantly elevated in αMan2 knockout (KO) mosquitoes, but there were complex interactions with virus infection and replication. In Wolbachia‐uninfected mosquitoes, the αMan2 KO mutation was associated with decreased DENV titres, but in a Wolbachia‐infected background, the αMan2 KO mutation significantly increased virus titres. In contrast, the αMan2 KO mutation significantly increased MAYV replication in Wolbachia‐uninfected mosquitoes and did not affect Wolbachia‐mediated virus blocking. These results demonstrate that αMan2 modulates arbovirus infection in A. aegypti mosquitoes in a pathogen‐ and Wolbachia‐specific manner, and that Wolbachia‐mediated pathogen blocking is a complex phenotype dependent on the mosquito host genotype and the pathogen. These results have a significant impact for the design and use of Wolbachia‐based strategies to control vector‐borne pathogens. [ABSTRACT FROM AUTHOR]

In this paper, we explore and study a mosquito population suppression model, consisting of two sub-models switching each other, based on Wolbachia-infected mosquitoes. Under the assumptions that the releases of Wolbachia-infected males are impulsive and periodic and the waiting period between two consecutive releases is not less than the sexually active lifespan of Wolbachia-infected males, we give a fairly complete description of the periodic dynamics of the model including the nonexistence of periodic solutions and the existence of a unique periodic solution and exact two periodic solutions. We also analyze the asymptotic stability of each periodic solution and each constant equilibrium in detail. Finally, some numerical examples are provided to illustrate our theoretical results. [ABSTRACT FROM AUTHOR]

BACKGROUNDS: In order to provide a long‐lasting formulation for spinosad (SP) targeting larval stages of Aedes aegypti (Linnaeus) and others alike, a SP tablet was developed based on microspheres, using polylactic acid as inside coating material. The microspheres were encapsulated using polyethylene glycol and 1‐hexadecanol to form a sustained‐release SP tablet. Micromorphology, active ingredient loading, structure identification, photolysis resistance and biological activity were evaluated in this report. RESULTS: (i) The SP microspheres had an average particle size of 6.16 ± 2.28 μm, low adhesion and good dispersion as evaluated by scanning electron microscopy and morphology. (ii) The average active ingredient loading and encapsulation of SP microspheres were 32.80 ± 0.74% and 78.41 ± 2.22%, respectively. (iii) The chemical structure of encapsulated SP was confirmed by Fourier transform infrared and 1H‐nuclear magnetic resonance. (iv) The photostability of the microspheres and the tablets were evaluated. The results showed that DT50 (time required to dissipate 50% of the mass originally present) of SP was 0.95 days in microspheres and 6.94 days in tablets. (v) The long‐term insecticidal activity of SP tablets was investigated, and the tablet had a long‐lasting activity against the mosquito larvae, showing 100% larval mortality for 63 days. CONCLUSIONS: The study provided a new long‐lasting formulation of SP, which displayed good efficacy in the control of Ae. aegypti larvae. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Little is known about Native American adaptations to blood-sucking arthropods prior to and following European contact. Multiple accounts starting in the 16th century suggest that rancid animal grease was employed by Gulf Coast indigenes as a mosquito repellent. Although many Native American ethnobotanical remedies for biting insects have been recorded, the use of animal products for this purpose is not well documented. Moreover, few traditional Native American mosquito repellents have been examined using controlled laboratory methods for repellency testing. In this study, we tested the repellent efficacy of fats derived from alligator, bear, cod, and shark that were aged to various stages of rancidity. Using yellow fever mosquitoes, (Aedes aegypti), we performed an arm-in-cage assay to measure the complete protection times resulted from these fats, when applied to human skin. We used a Y-tube olfactometer assay to evaluate long-distance repellency and tested tick-repellency in a crawling assay. Our results suggest that rancid animal fats from cod, bear, and alligator are potent albeit short-lived mosquito repellents. We found that both rancid and fresh fats do not repel ticks. Our findings show the validity of traditional ethnozoological knowledge of Native American people and support aspects of the ethnohistorical record. [ABSTRACT FROM AUTHOR]

Due to limitations in conventional disease vector control strategies including the rise of insecticide resistance in natural populations of mosquitoes, genetic control strategies using CRISPR gene drive systems have been under serious consideration. The identification of CRISPR target sites in mosquito populations is a key aspect for developing efficient genetic vector control strategies. While genome-wide Cas9 target sites have been explored in mosquitoes, a precise evaluation of target sites focused on coding sequence (CDS) is lacking. Additionally, target site polymorphisms have not been characterized for other nucleases such as Cas12a, which require a different DNA recognition site (PAM) and would expand the accessibility of mosquito genomes for genetic engineering. We undertook a comprehensive analysis of potential target sites for both Cas9 and Cas12a nucleases within the genomes of natural populations of Anopheles gambiae and Aedes aegypti from multiple continents. We demonstrate that using two nucleases increases the number of targets per gene. Also, we identified differences in nucleotide diversity between North American and African Aedes populations, impacting the abundance of good target sites with a minimal degree of polymorphisms that can affect the binding of gRNA. Lastly, we screened for gRNAs targeting sex-determination genes that could be widely applicable for developing field genetic control strategies. Overall, this work highlights the utility of employing both Cas9 and Cas12a nucleases and underscores the importance of designing universal genetic strategies adaptable to diverse mosquito populations. [ABSTRACT FROM AUTHOR]

Over the past 50 years, there has been a marked increase in diseases like dengue fever, chikungunya, and Zika. The World Mosquito Program (WMP) has developed an approach that, instead of attempting to eliminate vector species, introduces Wolbachia into native Aedes aegypti populations through the release of Wolbachia-infected mosquitoes. Using this approach, a randomized controlled study recently demonstrated a 77% reduction in dengue across a treatment area within Yogyakarta, Indonesia. Existing release methods use the ground-based release of mosquito eggs or adults that are labor-intensive, are logistically challenging to scale up, and can be restrictive in areas where staff safety is a concern. To overcome these limitations, we developed a fully automated mosquito dosing release system that released smaller cohorts of mosquitoes over a wide area and integrated it into an uncrewed aerial vehicle. We established the effectiveness of this system using an aerial mark, release, and recapture approach. We then demonstrated that using only the aerial release method, we can establish Wolbachia infection in a naive Ae. aegypti population. In both cases, the use of aerial releases demonstrated comparable outcomes to ground-based releases without the required labor or risk. These two trials demonstrated the feasibility of using an aerial release approach for large-scale mosquito releases. Editor's summary: The World Mosquito Program recently developed a technique to control dengue transmission by releasing Wolbachia-infected Aedes aegypti mosquitoes. Wolbachia bacteria act as a virus-blocking agent and are maternally transmitted to the local mosquito populations. However, scaling ground-based releases to large geographic areas poses a challenge. To address this, Lin et al. developed an automated mosquito dosing release system and incorporated it into an uncrewed aerial vehicle (UAV). This system, which includes temperature and humidity control, is designed to release 150 mosquitos per dose. Two successful field trials in Fiji demonstrated that the UAV-based release achieved similar uniformity to ground release methods and successfully established Wolbachia infections in the native population over a 2-km2 area. —Melisa Yashinski [ABSTRACT FROM AUTHOR]

Effective mosquito population suppression has been repeatedly demonstrated in field trials through the release of male mosquitoes to induce sterile mating with wild females using the incompatible insect technique (IIT), the sterile insect technique (SIT), or their combination. However, upscaling these techniques requires a highly efficient and scalable approach for the sex separation of mass-reared mosquitoes to minimize the unintentional release of females, which can lead to either population replacement or biting nuisance, a major bottleneck up to now. Here, we report the successful development of an automated mosquito pupa sex sorter that can effectively separate large numbers of males from females for population suppression of Aedes aegypti, A. albopictus, and Culex quinquefasciatus. The male production capacity of the automated sex sorter was increased by ~17-fold compared with manual sex separation with the Fay-Morlan sorter and enabled one person to separate 16 million males per week. With ~0.5% female contamination, the produced males exhibited high flight ability and mating performance. The field trial demonstrates that the quality of A. albopictus males produced using the automated sex sorter is suitable for inducing population suppression. These results indicate that the automated sex sorter offers the potential to upscale IIT and SIT against mosquito vectors for disease control. Editor's summary: Several techniques have been developed to fight mosquito-borne diseases, including the release of sterile or incompatible male mosquitoes into the wild to reduce population growth. However, sorting mosquitoes by sex is a laborious process. Gong et al. have now developed an automated mosquito pupa sex sorter, which was tested on three mosquito species and shown to increase the production of males 17-fold when compared with a manual separation process. A field trial in Guangzhou, China showed that the process was capable of producing sufficient incompatible males without reducing their quality, which led to the suppression of wild populations of mosquitoes. —Amos Matsiko [ABSTRACT FROM AUTHOR]

Female mosquitoes produce eggs in gonadotrophic cycles that are divided between a previtellogenic and vitellogenic phase. Previtellogenic females consume water and sugar sources like nectar while also being attracted to hosts for blood feeding. Consumption of a blood meal activates the vitellogenic phase, which produces mature eggs and suppresses host attraction. In this study, we tested the hypothesis that neuropeptide Y-like hormones differentially modulate host attraction behavior in the mosquito Aedes aegypti. A series of experiments collectively indicated that enteroendocrine cells (EECs) in the posterior midgut produce and release neuropeptide F (NPF) into the hemolymph during the previtellogenic phase which stimulates attraction to humans and biting behavior. Consumption of a blood meal, which primarily consists of protein by dry weight, down-regulated NPF in EECs until mature eggs developed, which was associated with a decline in hemolymph titer. NPF depletion depended on protein digestion but was not associated with EEC loss. Other experiments showed that neurons in the terminal ganglion extend axons to the posterior midgut and produce RYamide, which showed evidence of increased secretion into circulation after a blood meal. Injection of RYamide-1 and -2 into previtellogenic females suppressed host attraction, while coinjection of RYamides with or without short NPF-2 also inhibited the host attraction activity of NPF. Overall, our results identify NPF and RYamide as gut-associated hormones in A. aegypti that link host attraction behavior to shifts in diet during sequential gonadotrophic cycles. [ABSTRACT FROM AUTHOR]

Background: The spread of mosquito-transmitted diseases such as dengue is a major public health issue worldwide. The Aedes aegypti mosquito, a primary vector for dengue, thrives in urban environments and breeds mainly in artificial or natural water containers. While the relationship between urban landscapes and potential breeding sites remains poorly understood, such a knowledge could help mitigate the risks associated with these diseases. This study aimed to analyze the relationships between urban landscape characteristics and potential breeding site abundance and type in cities of French Guiana (South America), and to evaluate the potential of such variables to be used in predictive models. Methods: We use Multifactorial Analysis to explore the relationship between urban landscape characteristics derived from very high resolution satellite imagery, and potential breeding sites recorded from in-situ surveys. We then applied Random Forest models with different sets of urban variables to predict the number of potential breeding sites where entomological data are not available. Results: Landscape analyses applied to satellite images showed that urban types can be clearly identified using texture indices. The Multiple Factor Analysis helped identify variables related to the distribution of potential breeding sites, such as buildings class area, landscape shape index, building number, and the first component of texture indices. Models predicting the number of potential breeding sites using the entire dataset provided an R² of 0.90, possibly influenced by overfitting, but allowing the prediction over all the study sites. Predictions of potential breeding sites varied highly depending on their type, with better results on breeding sites types commonly found in urban landscapes, such as containers of less than 200 L, large volumes and barrels. The study also outlined the limitation offered by the entomological data, whose sampling was not specifically designed for this study. Model outputs could be used as input to a mosquito dynamics model when no accurate field data are available. Conclusion: This study offers a first use of routinely collected data on potential breeding sites in a research study. It highlights the potential benefits of including satellite-based characterizations of the urban environment to improve vector control strategies. [ABSTRACT FROM AUTHOR]

Chironomid (Diptera: Chironomidae) larvae play a key role in aquatic food webs as prey for predators like amphibian and dragonfly larvae. This trophic link may be disrupted by anthropogenic stressors such as Bacillus thuringiensis var. israelensis (Bti), a biocide widely used in mosquito control. In a companion study, we recorded a 41% reduction of non-target larval chironomids abundance in outdoor floodplain pond mesocosms (FPMs) treated with Bti. Therefore, we examined the diet of two top predators in the FPMs, larvae of the palmate newt (Salamandridae: Lissotriton helveticus) and dragonfly (Aeshnidae: predominantly Anax imperator), using bulk stable isotope analyses of carbon and nitrogen. Additionally, we determined neutral lipid fatty acids in newt larvae to assess diet-related effects on their physiological condition. We did not find any effects of Bti on the diet proportions of newt larvae and no significant effects on the fatty acid content. We observed a trend in Aeshnidae larvae from Bti-FPMs consuming a higher proportion of large prey (Aeshnidae, newt, damselfly larvae; ~42%), and similar parts of smaller prey (chironomid, mayfly, Libellulidae, and zooplankton), compared to controls. Our findings may suggest bottom-up effects of Bti on aquatic predators but should be further evaluated, for instance, by using compound-specific stable isotope analyses of fatty acids or metabarcoding approaches. [ABSTRACT FROM AUTHOR]

Increasing reports of insecticide resistance continue to hamper the gains of vector control strategies in curbing malaria transmission. This makes identifying new insecticide targets or alternative vector control strategies necessary. CLassifier of Essentiality AcRoss EukaRyote (CLEARER), a leave-one-organism-out cross-validation machine learning classifier for essential genes, was used to predict essential genes in Anopheles gambiae and selected predicted genes experimentally validated. The CLEARER algorithm was trained on six model organisms: Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens, Mus musculus, Saccharomyces cerevisiae and Schizosaccharomyces pombe, and employed to identify essential genes in An. gambiae. Of the 10,426 genes in An. gambiae, 1,946 genes (18.7%) were predicted to be Cellular Essential Genes (CEGs), 1716 (16.5%) to be Organism Essential Genes (OEGs), and 852 genes (8.2%) to be essential as both OEGs and CEGs. RNA interference (RNAi) was used to validate the top three highly expressed non-ribosomal predictions as probable vector control targets, by determining the effect of these genes on the survival of An. gambiae G3 mosquitoes. In addition, the effect of knockdown of arginase (AGAP008783) on Plasmodium berghei infection in mosquitoes was evaluated, an enzyme we computationally inferred earlier to be essential based on chokepoint analysis. Arginase and the top three genes, AGAP007406 (Elongation factor 1-alpha, Elf1), AGAP002076 (Heat shock 70kDa protein 1/8, HSP), AGAP009441 (Elongation factor 2, Elf2), had knockdown efficiencies of 91%, 75%, 63%, and 61%, respectively. While knockdown of HSP or Elf2 significantly reduced longevity of the mosquitoes (p<0.0001) compared to control groups, Elf1 or arginase knockdown had no effect on survival. However, arginase knockdown significantly reduced P. berghei oocytes counts in the midgut of mosquitoes when compared to LacZ-injected controls. The study reveals HSP and Elf2 as important contributors to mosquito survival and arginase as important for parasite development, hence placing them as possible targets for vector control. [ABSTRACT FROM AUTHOR]

Background: Aedes and Anopheles mosquitoes are responsible for tremendous global health burdens from their transmission of pathogens causing malaria, lymphatic filariasis, dengue, and yellow fever. Innovative vector control strategies will help to reduce the prevalence of these diseases. Mass rearing of mosquitoes for research and support of these strategies presently depends on meals of vertebrate blood, which is subject to acquisition, handling, and storage issues. Various blood-free replacements have been formulated for these mosquitoes, but none of these replacements are in wide use, and little is known about their potential impact on competence of the mosquitoes for Plasmodium infection. Methods: Colonies of Aedes aegypti and Anopheles stephensi were continuously maintained on a blood-free replacement (SkitoSnack; SS) or bovine blood (BB) and monitored for engorgement and hatch rates. Infections of Ae. aegypti and An. stephensi were assessed with Plasmodium gallinaceum and P. falciparum, respectively. Results: Replicate colonies of mosquitoes were maintained on BB or SS for 10 generations of Ae. aegypti and more than 63 generations of An. stephensi. The odds of engorgement by SS- relative to BB-maintained mosquitoes were higher for both Ae. aegypti (OR = 2.6, 95% CI 1.3–5.2) and An. stephensi (OR 2.7, 95% CI 1.4–5.5), while lower odds of hatching were found for eggs from the SS-maintained mosquitoes of both species (Ae. aegypti OR = 0.40, 95% CI 0.26–0.62; An. stephensi OR = 0.59, 95% CI 0.36–0.96). Oocyst counts were similar for P. gallinaceum infections of Ae. aegypti mosquitoes maintained on SS or BB (mean ratio = [mean on SS]/[mean on BB] = 1.11, 95% CI 0.85–1.49). Similar oocyst counts were also observed from the P. falciparum infections of SS- or BB-maintained An. stephensi (mean ratio = 0.76, 95% CI 0.44–1.37). The average counts of sporozoites/mosquito showed no evidence of reductions in the SS-maintained relative to BB-maintained mosquitoes of both species. Conclusions: Aedes aegypti and An. stephensi can be reliably maintained on SS over multiple generations and are as competent for Plasmodium infection as mosquitoes maintained on BB. Use of SS alleviates the need to acquire and preserve blood for mosquito husbandry and may support new initiatives in fundamental and applied research, including novel manipulations of midgut microbiota and factors important to the mosquito life cycle and pathogen susceptibility. [ABSTRACT FROM AUTHOR]

Global 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. [ABSTRACT FROM AUTHOR]

Wolbachia continue to be reported in species previously thought to lack them, particularly Aedes aegypti mosquitoes. The presence of Wolbachia in this arbovirus vector is considered important because releases of mosquitoes with transinfected Wolbachia are being used around the world to suppress pathogen transmission and these efforts depend on a lack of Wolbachia in natural populations of this species. We previously assessed papers reporting Wolbachia in natural populations of Ae. aegypti and found little evidence that seemed convincing. However, since our review, more and more papers are emerging on Wolbachia detections in this species. Our purpose here is to evaluate these papers within the context of criteria we previously established but also new criteria that include the absence of releases of transinfections within the local areas being sampled which has contaminated natural populations in at least one case where novel detections have been reported. We also address the broader issue of Wolbachia detection in other insects where similar issues may arise which can affect overall estimates of this endosymbiont more generally. We note continuing shortcomings in papers purporting to find natural Wolbachia in Ae. aegypti which are applicable to other insects as well. [ABSTRACT FROM AUTHOR]

The dengue vector Aedes aegypti is exceptionally threatening worldwide as it causes a primary viral disease (dengue), even though the vector species has other viral loads. This study was aimed at investigating the Annona reticulata extract with respect to the larvicidal activity against the 4th instar larvae of Aedes aegypti, the toxicity on a non-target aquatic organism (Chironomus costatus), and the biochemical marker enzymes in the mosquito larvae. Among various solvent tested, ethyl acetate as extraction solvent showed the highest mortality (100%) at 200 ppm against Aedes aegypti with no adverse effect on the non-target aquatic organism tested. Besides, the ethyl acetate extract showed significant inhibition of acetylcholinesterase (19%), α- and β-carboxylesterases (47.2 & 70.3%, respectively), glutathione S-transferase (35.2%), and alkaline phosphatase (22.3%). Moreover, the protein (13.7%) and acid phosphatase (11.2%) were reduced without being significant in the quantitative analyses. In qualitative analyses, we also observed that all enzyme patterns showed differential expression upon exposure to the ethyl acetate extract against the 4th instar larvae of Aedes aegypti at a lethal concentration (LC50: 72.5 ppm) for 24 h. These findings precisely showed that the Annona reticulata active extract (using ethyl acetate) can be used as an alternative in pest and vector control management contributing towards not spreading infectious diseases. [ABSTRACT FROM AUTHOR]