Population genetics of Aedes albopictus in the port cities of Hainan Island and Leizhou Peninsula, China.

BACKGROUND: Aedes albopictus is an important vector of arboviral diseases, transmitting yellow fever, dengue fever, chikungunya and Zika. Monitoring its population genetic diversity and genetic differentiation has become essential for the control of infectious disease epidemics, especially in the functional areas of ports of entry. Population genetic monitoring of Ae. albopictus in the port area can help in the monitoring of port mosquito invasions and establishing port sanitary and quarantine measures to prevent the introduction and transmission of vector-borne diseases. METHODS: Seventeen populations of Ae. albopictus were collected from five port cities on Hainan Island and the Leizhou Peninsula, 8 populations were collected from port areas, 4 from urban areas and 5 from rural areas. Nine microsatellite loci and the mitochondrial COI gene were used to study the population genetic diversity, population genetic structure and interpopulation gene flow of Ae. albopictus. RESULTS: The nine microsatellite loci used were highly polymorphic, with an average PIC value of 0.768. The UPGMA genetic tree, STRUCTURE barplot and PCoA analyses showed that the 17 Ae. albopictus populations could be divided into three genetic groups. All 17 populations showed high haplotype diversity (Hd = 0.8069-0.9678) and formed 133 distinct haplotypes. These haplotypes can be divided into four genetic clades, but they are not associated with the geographical distribution of Ae. albopictus. Fst and Nm showed strong gene flow and little differentiation among populations. CONCLUSION: Ae. albopictus in port areas are not significantly different from urban and rural populations due to strong gene flow, which prevents differentiation and increases the genetic diversity of the populations. High genetic diversity facilitates mosquito adaptation to complex environmental changes, which is a challenge for vector-borne disease control in port areas.

Evolution of knockdown resistance (kdr) mutations of Aedes aegypti and Aedes albopictus in Hainan Island and Leizhou Peninsula, China.

Background: Aedes aegypti and Aedes albopictus are important vectors of human arboviruses, transmitting arboviral diseases such as yellow fever, dengue, chikungunya and Zika. These two mosquitoes coexist on Hainan Island and the Leizhou Peninsula in China. Over the past 40 years, the distribution of Ae. albopictus has gradually expanded in these areas, while the distribution of Ae. aegypti has declined dramatically mainly due to the ecological changes and some other factors such as heavy use of insecticide indoor based on endophagic bloodfeeding of the species. Methods: This study focused on the knockdown resistance (kdr) genes of both mosquitoes, investigated their mutations, and analyzed their haplotype and evolutionary diversity combined with population genetic features based on the ND4/ND5 genes to further elucidate the molecular mechanisms underlying the development of insecticide resistance in both mosquitoes. Results: Three mutations, S989P, V1016G and F1534C, were found to be present in Ae. aegypti populations, and the three mutations occurred synergistically. Multiple mutation types (F1534C/S/L/W) of the F1534 locus are found in Ae. albopictus populations, with the three common mutations F1534C, F1534S and F1534L all having multiple independent origins. The F1534W (TTC/TGG) mutation is thought to have evolved from the F1534L (TTC/TTG) mutation. The F1534S (TTC/TCG) mutation has evolved from the F1534S (TTC/TCC) mutation. The most common form of mutation at the F1534 locus found in this study was S1534C, accounting for 20.97%, which may have evolved from the F1534C mutation. In addition, a new non-synonymous mutation M1524I and 28 synonymous mutations were identified in Ae. albopictus populations. Correlation analysis showed that the genetic diversity of Ae. aegypti and Ae. albopictus populations did not correlate with their kdr haplotype diversity (P>0.05), but strong gene flow between populations may have contributed to the evolution of the kdr gene. Conclusion: The study of kdr gene evolution in the two mosquito species may help to identify the evolutionary trend of insecticide resistance at an early stage and provide a theoretical basis for improving the efficiency of biological vector control and subsequent research into new insecticides.

First Report of Rubber Collection Bowls & Plastic and Bamboo Water Containers as the Major Breeding Source of Ae. albopictus with the Indigenous Transmission of Dengue and Chikungunya in Rural Forested Malaria-Endemic Villages of Dhalai District, Tripura, India: The Importance of Molecular Identification.

BACKGROUND: With the reports of indigenous cases of dengue and chikungunya in the forest-covered rural tribal malaria-endemic villages of Dhalai District, Tripura, India, an exploratory study was undertaken to identify the vector breeding sites. METHODS: From June 2021 to August 2022, mosquito larvae were collected from both natural and artificial sources in the villages, house premises, and their nearby forested areas outside of the houses. Other than morphological characterisation, Aedes species were confirmed by polymerase chain reaction targeting both nuclear (ITS2) and mitochondrial genes (COI) followed by bidirectional Sanger sequencing. RESULTS: Aedes albopictus was abundantly found in this area in both natural and artificial containers, whereas Ae. aegypti was absent. Among the breeding sources of molecularly confirmed Ae. albopictus species, rubber collection bowls were found to be a breeding source reported for the first time. Plastic and indigenously made bamboo-polythene containers for storing supply water and harvesting rainwater in the villages with a shortage of water were found to be other major breeding sources, which calls for specific vector control strategies. Natural sources like ponds and rainwater collected on Tectona grandis leaves and Colocasia axil were also found to harbour the breeding, along with other commonly found sources like bamboo stumps and tree holes. No artificial containers as a breeding source were found inside the houses. Mixed breeding was observed in many containers with other Aedes and other mosquito species, necessitating molecular identification. We report six haplotypes in this study, among which two are reported for the first time. However, Aedes aegypti was not found in the area. Additionally, rubber collection bowls, ponds, and water containers also showed the presence of Culex quinquefasciatus and Culex vishnui, known JE vectors from this area, and reported JE cases as well. Different Anopheles vector spp. from this known malaria-endemic area were also found, corroborating this area as a hotbed of several vectors and vector-borne diseases. CONCLUSIONS: This study, for the first time, reports the breeding sources of Aedes albopictus in the forested areas of Tripura, with rubber collection bowls and large water storage containers as major sources. Also, for the first time, this study reports the molecular characterisation of the Ae. albopictus species of Tripura, elucidating the limitations of morphological identification and highlighting the importance of molecular studies for designing appropriate vector control strategies. The study also reports the co-breeding of JE and malaria vectors for the first time in the area reporting these vector-borne diseases.

Gut Bacterial Diversity of Field and Laboratory-Reared Aedes albopictus Populations of Rio de Janeiro, Brazil.

BACKGROUND: The mosquito microbiota impacts different parameters in host biology, such as development, metabolism, immune response and vector competence to pathogens. As the environment is an important source of acquisition of host associate microbes, we described the microbiota and the vector competence to Zika virus (ZIKV) of Aedes albopictus from three areas with distinct landscapes. METHODS: Adult females were collected during two different seasons, while eggs were used to rear F1 colonies. Midgut bacterial communities were described in field and F1 mosquitoes as well as in insects from a laboratory colony (>30 generations, LAB) using 16S rRNA gene sequencing. F1 mosquitoes were infected with ZIKV to determine virus infection rates (IRs) and dissemination rates (DRs). Collection season significantly affected the bacterial microbiota diversity and composition, e.g., diversity levels decreased from the wet to the dry season. Field-collected and LAB mosquitoes' microbiota had similar diversity levels, which were higher compared to F1 mosquitoes. However, the gut microbiota composition of field mosquitoes was distinct from that of laboratory-reared mosquitoes (LAB and F1), regardless of the collection season and location. A possible negative correlation was detected between Acetobacteraceae and Wolbachia, with the former dominating the gut microbiota of F1 Ae. albopictus, while the latter was absent/undetectable. Furthermore, we detected significant differences in infection and dissemination rates (but not in the viral load) between the mosquito populations, but it does not seem to be related to gut microbiota composition, as it was similar between F1 mosquitoes regardless of their population. CONCLUSIONS: Our results indicate that the environment and the collection season play a significant role in shaping mosquitoes' bacterial microbiota.

Repellent Screening of Selected Plant Essential Oils Against Dengue Fever Mosquitoes Using Behavior Bioassays.

Among the efforts to reduce mosquito-transmitted diseases, such as malaria and dengue fever, essential oils (EOs) have become increasingly popular as natural replacements for the repellant DEET. In this study, seven commercially available plant EOs against Aedes species mosquitoes were evaluated for their complete protection time (CPT, min) in vivo using human-hand in cage tests (GB2009/China and WHO2009). Among the EOs with the highest efficacy in repelling mosquitoes, Aedes albopictus (Skuse) were clove bud oil and patchouli oil. Both were further assessed according to the in vivo method recommended by the WHO, to determine their minimum effective dose and CPT. A comparison of the ED50 values (dose yielding a 50% repellent response) of these two EOs against Aedes aegypti(L.) showed that the ED50 (2.496 µg/cm2) of patchouli oil was 1248 times higher than that of clove bud oil (0.002 µg/cm2), thus demonstrating them greater efficacy of the latter in repelling Ae. aegypti mosquitoes. For the 2 EOs, eugenol was the major component with higher than 80% in relative amount of the clove bud oil. The patchouli oil had more than 30% of character chemical patchouli alcohol along with α-bulnesene (10.962%), α-guaiene (9.227%), and seychellene (7.566%). Clove bud oil was found to confer longer complete protection than patchouli oil against a common species of mosquito. These results suggest use of EOs as safe, highly potent repellents for use in daily life and against mosquito-transmitted diseases, such as malaria and dengue fever.

Isobaric tags for relative and absolute quantification-based proteomic analysis of host-pathogen protein interactions in the midgut of Aedes albopictus during dengue virus infection.

Aedes albopictus (Ae. albopictus), an important vector of dengue virus (DENV), is distributed worldwide. Identifying host proteins involved in flavivirus replication in Ae. albopictus and determining their natural antiviral mechanisms are critical to control virus transmission. Revealing the key proteins related to virus replication and exploring the host-pathogen interaction are of great significance in finding new pathways of the natural immune response in Ae. albopictus. Isobaric tags for relative and absolute quantification (iTRAQ) was used to perform a comparative proteomic analysis between the midgut of Ae. albopictus infected with DENV and the control. 3,419 proteins were detected, of which 162 were ≥ 1.2-fold differentially upregulated or ≤ 0.8-fold differentially downregulated (p < 0.05) during DENV infections. Differentially expressed proteins (DEPs) were mainly enriched in ubiquitin ligase complex, structural constituent of cuticle, carbohydrate metabolism, and lipid metabolism pathways. We found that one of the DEPs, a putative pupal cuticle (PC) protein could inhibit the replication of DENV and interact with the DENV-E protein. In addition, the result of immunofluorescence (IF) test showed that there was co-localization between ubiquitin carboxyl-terminal hydrolase (UCH) protein and the DENV-E protein, and virus infection reduced the level of this protein. iTRAQ-based proteomic analysis of the Ae. albopictus midgut identified dengue infection-induced upregulated and downregulated proteins. The interaction between the PC and UCH proteins in the midgut of Ae. albopictus might exert a natural antiviral mechanism in mosquito.

Snow-Covered Tires Generate Microhabitats That Enhance Overwintering Survival of Aedes albopictus (Diptera: Culicidae) in the Midwest, USA.

The Asian tiger mosquito, Aedes albopictus (Skuse), is a public health threat because it can potentially transmit multiple pathogenic arboviruses, exhibits aggressive diurnal biting, and is highly invasive. As Ae. albopictus moved northward into the United States, the limits of expansion were predicted as locations with a mean January temperature warmer than -2.5°C. We postulated that the range of Ae. albopictus could exceed these temperature limits if eggs in diapause overwinter in tires that provide an insulating effect from extreme temperatures. Fifteen tires with Ae. albopictus and Aedes triseriatus (Say) eggs, a native cold hardy species, were placed outside at five locations along a latitudinal gradient in Wisconsin and Illinois during the winter of 2018-2019; notably, in January 2019, a regional arctic air event brought the lowest temperatures recorded in over 20 yr. External and internal tire temperatures were recorded at 3 hr intervals, and egg survival was recorded after six months. Aedes albopictus eggs survived only from tires at northernmost locations. The mean internal January temperature of tires that supported survival was -1.8°C, while externally the mean temperature was -5.3°C, indicating that tires provided an average of +3.5°C of insulation. Tires that supported egg survival also had over 100 mm of snow cover during January. In the absence of snow cover, tires across the study area provided an average +0.79°C [95% CI 0.34-1.11] insulation. This work provides strong argument for the inclusion of microhabitats in models of dispersal and establishment of Ae. albopictus and other vector species.

Maternally Instigated Diapause in Aedes albopictus: Coordinating Experience and Internal State for Survival in Variable Environments.

The Asian tiger mosquito, Aedes albopictus, is one of the most dangerous invasive species in the world. Females bite mammalian hosts, including humans, to obtain blood for egg development. The ancestral range of Ae. albopictus likely spanned from India to Japan and this species has since invaded a substantial portion of the globe. Ae. albopictus can be broadly categorized into temperate and tropical populations. One key to their ability to invade diverse ecological spaces is the capacity of females to detect seasonal changes and produce stress-resistant eggs that survive harsh winters. Females living in temperate regions respond to cues that predict the onset of unfavorable environmental conditions by producing eggs that enter maternally instigated embryonic diapause, a developmentally arrested state, which allows species survival by protecting the embryos until favorable conditions return. To appropriately produce diapause eggs, the female must integrate environmental cues and internal physiological state (blood feeding and reproductive status) to allocate nutrients and regulate reproduction. There is variation in reproductive responses to environmental cues between interfertile tropical and temperate populations depending on whether females are actively producing diapause vs. non-diapause eggs and whether they originate from populations that are capable of diapause. Although diapause-inducing environmental cues and diapause eggs have been extensively characterized, little is known about how the female detects gradual environmental changes and coordinates her reproductive status with seasonal dynamics to lay diapause eggs in order to maximize offspring survival. Previous studies suggest that the circadian system is involved in detecting daylength as a critical cue. However, it is unknown which clock network components are important, how these connect to reproductive physiology, and how they may differ between behavioral states or across populations with variable diapause competence. In this review, we showcase Ae. albopictus as an emerging species for neurogenetics to study how the nervous system combines environmental conditions and internal state to optimize reproductive behavior. We review environmental cues for diapause induction, downstream pathways that control female metabolic changes and reproductive capacity, as well as diapause heterogeneity between populations with different evolutionary histories. We highlight genetic tools that can be implemented in Ae. albopictus to identify signaling molecules and cellular circuits that control diapause. The tools and discoveries made in this species could translate to a broader understanding of how environmental cues are interpreted to alter reproductive physiology in other species and how populations with similar genetic and circuit organizations diversify behavioral patterns. These approaches may yield new targets to interfere with mosquito reproductive capacity, which could be exploited to reduce mosquito populations and the burden of the pathogens they transmit.

Recombinant Mosquito Densovirus with Bti Toxins Significantly Improves Pathogenicity against Aedes albopictus.

Mosquito densoviruses (MDVs) are mosquito-specific viruses that are recommended as mosquito bio-control agents. The MDV Aedes aegypti densovirus (AeDNV) is a good candidate for controlling mosquitoes. However, the slow activity restricts their widespread use for vector control. In this study, we introduced the Bacillus thuringiensis (Bti) toxin Cry11Aa domain II loop α8 and Cyt1Aa loop ß6-αE peptides into the AeDNV genome to improve its mosquitocidal efficiency; protein expression was confirmed using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS). Recombinant plasmids were transfected into mosquito C6/36 cell lines, and the expression of specific peptides was detected through RT-PCR. A toxicity bioassay against the first instar Aedes albopictus larvae revealed that the pathogenic activity of recombinant AeDNV was significantly higher and faster than the wild-type (wt) viruses, and mortality increased in a dose-dependent manner. The recombinant viruses were genetically stable and displayed growth phenotype and virus proliferation ability, similar to wild-type AeDNV. Our novel results offer further insights by combining two mosquitocidal pathogens to improve viral toxicity for mosquito control.

High Prevalence of Rickettsia bellii in Mosquitoes From Eastern China.

Mosquitoes are the most important vectors carrying significant numbers of human pathogens. Recent studies implicated that mosquitoes play an important role in circulation and transmission of multiple Rickettsia species. In this study, Rickettsia bellii was identified in four mosquito species (Culex pipiens, C. tritaeniorhynchus, Aedes albopictus, and Anopheles sinensis) collected from three Eastern China provinces during 2019-2020. Rickettsia bellii was detected in 37.50 and 26.32% of the C. pipiens pools from Beijing and Jiangsu province, respectively. In C. tritaeniorhynchus and An. sinensis from Shandong, the infection rate is 20.00 and 6.25%, respectively. Additionally, three Ae. albopictus pools (3/42, 7.14%) from Beijing were also detected positive for R. bellii. Genetic and phylogenetic analysis on 16S, gltA, and groEL genes indicates that sequences from all these strains are highly homologous and closely related to other R. bellii strains. This is the first report that Ae. albopictus and C. tritaeniorhynchus harbor R. bellii. The wide host range and high infection rate in certain areas may dramatically increase the exposure of R. bellii to human and other vertebrates. The role of mosquitoes in transmission of rickettsiosis and its potential risk to public health should be further considered.

Bioassay studies on the reaction of Aedes aegypti & Aedes albopictus (Diptera: Culicidae) on different attractants.

BACKGROUND: The studies on mosquito attractants had been develop intensively in the recent years. However, the study on fruit peel extract as mosquito attractant was scarce, even though various fruits had demonstrated the ability to attract different types of mosquito species. OBJECTIVE: This study aims to determine the potential of Carica papaya (papaya) and Ananas comosus (pineapple) peel extracts to attract Aedes albopictus and Aedes aegypti. METHODS: The Aedes mosquitoes response to the fruit peel extracts were conducted in the no-choice and choice assay using modified olfactometer. The Preference Index (PI) in each assay was calculated and arcsine transformed before conducting independent t-test to determine the significant different between the mean arcsine transformed PI and the tested hypothesis mean PI. RESULT: No choice assay indicate both Aedes species have significant attraction to the papaya and pineapple peel extracts (p < 0.05). In choice assay, Ae. albopictus is revealed to equally attracted to the papaya and pineapple peel extracts (p > 0.05) while Ae. aegypti is significantly attracted to the papaya peel extract (p < 0.05). CONCLUSION: The study had identified that both fruit peel extracts able to attract Aedes mosquitoes with Ae. albopictus is equally attracted to papaya and pineapple peel extracts while Ae. aegypti is more attracted to the papaya peel extract than the pineapple peel extract.

Improved reference genome of the arboviral vector Aedes albopictus.

BACKGROUND: The Asian tiger mosquito Aedes albopictus is globally expanding and has become the main vector for human arboviruses in Europe. With limited antiviral drugs and vaccines available, vector control is the primary approach to prevent mosquito-borne diseases. A reliable and accurate DNA sequence of the Ae. albopictus genome is essential to develop new approaches that involve genetic manipulation of mosquitoes. RESULTS: We use long-read sequencing methods and modern scaffolding techniques (PacBio, 10X, and Hi-C) to produce AalbF2, a dramatically improved assembly of the Ae. albopictus genome. AalbF2 reveals widespread viral insertions, novel microRNAs and piRNA clusters, the sex-determining locus, and new immunity genes, and enables genome-wide studies of geographically diverse Ae. albopictus populations and analyses of the developmental and stage-dependent network of expression data. Additionally, we build the first physical map for this species with 75% of the assembled genome anchored to the chromosomes. CONCLUSION: The AalbF2 genome assembly represents the most up-to-date collective knowledge of the Ae. albopictus genome. These resources represent a foundation to improve understanding of the adaptation potential and the epidemiological relevance of this species and foster the development of innovative control measures.

Population structure and genetic diversity of Aedes aegypti and Aedes albopictus in Penang as revealed by mitochondrial DNA cytochrome oxidase I.

The population genetics study is crucial as it helps in understanding the epidemiological aspects of dengue and help improving a vector control measures. This research aims to investigate the population genetics structure of two common species of Aedes mosquitoes in Penang; Aedes aegypti and Aedes albopictus using Cytochrome Oxidase I (COI) mitochondrial DNA (mtDNA) marker. Molecular investigations were derived from 440 bp and 418 bp mtDNA COI on 125 and 334 larvae of Aedes aegypti and Aedes albopictus respectively, from 32 locations in Penang. All samples were employed in the BLASTn for species identification. The haplotype diversity, nucleotide diversity, neutrality test and mismatch distribution analysis were conducted in DnaSP version 5.10.1. AMOVA analysis was conducted in ARLEQUIN version 3.5 and the phylogenetic reconstructions based on maximum likelihood (ML) and neighbor-joining (NJ) methods were implemented in MEGA X. The relationships among haplotypes were further tested by creating a minimum spanning tree using Network version 4.6.1. All samples were genetically identified and clustered into six distinct species. Among the species, Ae. albopictus was the most abundant (67.2%), followed by Ae. aegypti (25.2%) and the rest were counted for Culex sp. and Toxorhynchites sp. Both Ae. aegypti and Ae. albopictus show low nucleotide diversity (π) and high haplotype diversity (h), while the neutrality test shows a negative value in most of the population for both species. There are a total of 39 and 64 haplotypes recorded for Ae. aegypti and Ae. albopictus respectively. AMOVA analysis revealed that most of the variation occurred within population for both species. Mismatch distribution analysis showed bimodal characteristic of population differentiation for Ae. aegypti but Ae. albopictus showed unimodal characteristics of population differentiation. Genetic distance based on Tamura-Nei parameter showed low genetic divergent within population and high genetic divergent among population for both species. The maximum likelihood tree showed no obvious pattern of population genetic structure for both Ae. aegypti and Ae. albopictus from Penang and a moderate to high bootstrap values has supported this conclusion. The minimum spanning network for Ae. aegypti and Ae. albopictus showed five and three dominant haplotypes respectively, which indicates a mixture of haplotypes from the regions analysed. This study revealed that there is no population genetic structure exhibited by both Ae. aegypti and Ae. albopictus in Penang. Mutation has occurred rapidly in both species and this will be challenging in controlling the populations. However, further analysis needed to confirm this statement.

Development and calibration of a model for the potential establishment and impact of Aedes albopictus in Europe.

The Asian tiger mosquito (Aedes albopictus) is one of the most invasive disease vectors worldwide. The species is a competent vector of dengue, chikungunya, Zika viruses and other severe parasites and pathogens threatening human health. The capacity of this mosquito to colonize and establish in new areas (including temperate regions) is enhanced by its ability of producing diapausing eggs that survive relatively cold winters. The main drivers of population dynamics for this mosquito are water and air temperature and photoperiod. In this paper, we present a mechanistic model that predicts the potential distribution, abundance and activity of Asian tiger mosquito in Europe. The model includes a comprehensive description of: i) the individual life-history strategies, including diapause, ii) the influence of weather-driven individual physiological responses on population dynamics and iii) the density-dependent regulation of larval mortality rate. The model is calibrated using field data from several locations along an altitudinal gradient in the Italian Alps, which enabled accurate prediction of cold temperature effects on population abundance, including identification of conditions that prevent overwintering of the species. Model predictions are consistent with the most updated information on species' presence and absence. Predicted population abundance shows a clear south-north decreasing gradient. A similar yet less evident pattern in the activity of the species is also predicted. The model represents a valuable tool for the development of strategies aimed at the management of Ae. albopictus and for the implementation of effective control measures against vector-borne diseases in Europe.

Neglected Urban Villages in Current Vector Surveillance System: Evidences in Guangzhou, China.

: Numerous urban villages (UVs) with substandard living conditions that cause people to live there with vulnerability to health impacts, including vector-borne diseases such as dengue fever (DF), are major environmental and public health concerns in highly urbanized regions, especially in developing countries. It is necessary to explore the relationship between UVs and vector for effectively dealing with these problems. In this study, land-use types, including UVs, normal construction land (NCL), unused land (UL), vegetation, and water, were retrieved from the high-resolution remotely sensed imagery in the central area of Guangzhou in 2017. The vector density from May to October in 2017, including Aedes.albopictus(Ae.albopictus)'s Breteau index (BI), standard space index (SSI), and adult density index (ADI) were obtained from the vector surveillance system implemented by the Guangzhou Center for Disease Control and Prevention (CDC). Furthermore, the spatial and temporal patterns of vector monitoring sites and vector density were analyzed on a fine scale, and then the Geodetector tool was further employed to explore the relationships between vector density and land-use types. The monitoring sites were mainly located in NCL (55.70%-56.44%) and UV (13.14%-13.92%). Among the total monitoring sites of BI (79), SSI (312), and ADI (326), the random sites accounted for about 88.61%, 97.12%, and 98.47%, respectively. The density of Ae.albopictus was temporally related to rainfall and temperature and was obviously differentiated among different land-use types. Meanwhile, the grids with higher density, which were mostly concentrated in the Pearl River fork zone that collects a large number of UVs, showed that the density of Ae.albopictus was spatially associated with the UVs. Next, the results of the Geodetector illustrated that UVs posed great impact on the density of Ae.albopictus across the central region of Guangzhou. We suggest that the number of monitoring sites in the UVs should be appropriately increased to strengthen the current vector surveillance system in Guangzhou. This study will provide targeted guidance for local authorities, making more effective control and prevention measures on the DF epidemics.

Vertical transmission of dengue virus in Aedes aegypti and its role in the epidemiological persistence of dengue in Central and Southern Mexico.

BACKGROUND: Aedes aegypti and Aedes albopictus are the main mosquito species responsible for dengue virus (DENV) transmission to humans in the tropical and subtropical regions of the world. The role of vertical transmission in the epidemiology of dengue and the maintenance of this arbovirus in nature during interepidemic periods remain poorly understood, and DENV vertical transmission could sustain the existence of virus reservoirs within Aedes populations. METHODS: Between April 2011 and October 2012, we monitored vertical transmission of DENV in Ae. aegypti and Ae. albopictus in 9 cities of 4 Mexican states. Aedes eggs were collected in ovitraps, then adults were reared under laboratory conditions and their heads were used to infect C6/36 cells. The presence of flavivirus was detected by immunofluorescence assays (IFA), and DENV infection was confirmed by RT-PCR. RESULTS: About 96% of reared adults were Ae. aegypti and 4.0% were Ae. albopictus. No infection was detected in Ae. albopictus, whereas 54 of 713 (7.8%) of Ae. aegypti pools tested positive. A minimum infection rate (MIR) of 2.52 per 1000 mosquitoes was estimated for Ae. aegypti. DENV-1, DENV-2 & DENV-3 serotypes were detected even during interepidemic periods. CONCLUSIONS: This study reports the evidence of vertical transmission of dengue virus with viral isolation and molecular confirmation in Ae. aegypti eggs collected in four endemic regions of Central and Southern Mexico. Vertical transmission may play a role as a reservoir mechanism during mosquito dormancy in interepidemic periods but with minor participation in transmission during epidemic periods.

TRANSMISSION VERTICALE DU VIRUS DE LA DENGUE CHEZ AEDES AEGYPTI ET SON RÔLE DANS LA PERSISTANCE ÉPIDÉMIOLOGIQUE DE LA DENGUE DANS LE CENTRE ET LE SUD DU MEXIQUE: OBJECTIF: Aedes aegypti et Aedes albopictus sont les principales espèces de moustiques responsables de la transmission du virus de la dengue (DENV) à l'homme dans les régions tropicales et subtropicales du monde. Le rôle de la transmission verticale dans l'épidémiologie de la dengue et le maintien de cet arbovirus dans la nature pendant les périodes d'inter-épidémiques restent mal compris, et la transmission verticale du DENV pourrait maintenir l'existence de réservoirs de virus au sein des populations d'Aedes. Notre objectif était d'évaluer la transmission verticale du DENV au Mexique. MÉTHODES: Entre avril 2011 et octobre 2012, nous avons surveillé la transmission verticale du DENV chez Ae. aegypti et Ae. albopictus dans 9 villes de 4 états mexicains. Les œufs d'Aedes ont été collectés dans des ovitraps, puis les adultes ont été élevés dans des conditions de laboratoire et leur tête a été utilisée pour infecter les cellules C6/36. La présence de flavivirus a été détectée par des tests d'immunofluorescence (IFA) et l'infection par DENV a été confirmée par RT-PCR. RÉSULTATS: 96% des adultes élevés étaient Ae. aegypti et 4,0% étaient Ae. albopictus. Aucune infection n'a été détectée chez Ae. albopictus, alors que 54 des 713 (7,8%) des pools d'Ae. aegypti ont été testés positifs. Un taux d'infection minimum (MIR) de 2,52 pour 1000 moustiques a été estimé pour Ae. aegypti. Les sérotypes DENV-1, DENV-2 et DENV-3 ont été détectés même pendant les périodes inter-épidémiques. CONCLUSIONS: Cette étude rapporte les preuves de transmission verticale du virus de la dengue avec isolement viral et confirmation moléculaire dans les œufs d'Ae. Aegypti collectés dans quatre régions d'endémie du centre et du sud du Mexique. La transmission verticale pourrait jouer un rôle de mécanisme réservoir lors de la dormance des moustiques en période inter-épidémique, mais avec une participation mineure à la transmission en période d'épidémie.

Applying fuzzy logic to assess the biogeographical risk of dengue in South America.

BACKGROUND: Over the last decade, reports about dengue cases have increase worldwide, which is particularly worrisome in South America due to the historic record of dengue outbreaks from the seventeenth century until the first half of the twentieth century. Dengue is a viral disease that involves insect vectors, namely Aedes aegypti and Ae. albopictus, which implies that, to prevent and combat outbreaks, it is necessary to understand the set of ecological and biogeographical factors affecting both the vector species and the virus. METHODS: We contribute with a methodology based on fuzzy logic that is helpful to disentangle the main factors that determine favorable environmental conditions for vectors and diseases. Using favorability functions as fuzzy logic modelling technique and the fuzzy intersection, union and inclusion as fuzzy operators, we were able to specify the territories at biogeographical risk of dengue outbreaks in South America. RESULTS: Our results indicate that the distribution of Ae. aegypti mostly encompasses the biogeographical framework of dengue in South America, which suggests that this species is the principal vector responsible for the geographical extent of dengue cases in the continent. Nevertheless, the intersection between the favorability for dengue cases and the union of the favorability for any of the vector species provided a comprehensive map of the biogeographical risk for dengue. CONCLUSIONS: Fuzzy logic is an appropriate conceptual and operational tool to tackle the nuances of the vector-illness biogeographical interaction. The application of fuzzy logic may be useful in decision-making by the public health authorities to prevent, control and mitigate vector-borne diseases.

Arbovirus surveillance: first dengue virus detection in local Aedes albopictus mosquitoes in Europe, Catalonia, Spain, 2015.

Dengue has emerged as the most important viral mosquito-borne disease globally. The current risk of dengue outbreaks in Europe appeared with the introduction of the vector Aedes albopictus mosquito in Mediterranean countries. Considering the increasing frequency of dengue epidemics worldwide and the movement of viraemic hosts, it is expected that new autochthonous cases will occur in the future in Europe. Arbovirus surveillance started in Catalonia in 2015 to monitor imported cases and detect possible local arboviral transmission. During 2015, 131 patients with a recent travel history to endemic countries were tested for dengue virus (DENV) and 65 dengue cases were detected. Twenty-eight patients with a febrile illness were viraemic, as demonstrated by a positive real-time RT-PCR test for DENV in serum samples. Entomological investigations around the viraemic cases led to the detection of DENV in a pool of local Ae. albopictus captured in the residency of one case. The sequence of the DENV envelope gene detected in the mosquito pool was identical to that detected in the patient. Our results show how entomological surveillance conducted around viraemic travellers can be effective for early detection of DENV in mosquitoes and thus might help to prevent possible autochthonous transmission.

Salinity tolerant Aedes aegypti and Ae. albopictus-Infection with dengue virus and contribution to dengue transmission in a coastal peninsula.

BACKGROUND & OBJECTIVES: Aedes aegypti and Ae. albopictus are major arboviral vectors that are considered to lay eggs, and undergo preimaginal development only in fresh water collections. However, recently they have been also shown to develop in coastal brackish water habitats. The ability of the biologically variant salinity-tolerant Aedes vectors to transmit arboviral diseases is unknown. We therefore, investigated the infection of salinity-tolerant Aedes mosquitoes with dengue virus (DENV) and analysed dengue incidence and rainfall data to assess the contribution of salinity-tolerant Aedes vectors to dengue transmission in the coastal Jaffna peninsula in Sri Lanka. METHODS: Brackish and fresh water developing female Ae. aegypti and Ae. albopictus were tested for their ability to become infected with DENV through in vitro blood feeding and then transmit DENV vertically to their progeny. An immunochromatographic test for the NS1 antigen was used to detect DENV. Temporal variation in dengue incidence in relation to rainfall was analysed for the peninsula and other parts of Sri Lanka. RESULTS: Aedes aegypti and Ae. albopictus developing in brackish water, became infected with DENV through in vitro blood feeding and the infected mosquitoes were able to vertically transmit DENV to their progeny. Monsoonal rainfall was the discernible factor responsible for the seasonal increase in dengue incidence in the peninsula and elsewhere in Sri Lanka. INTERPRETATION & CONCLUSION: Fresh water Aedes vectors are main contributors to the increased dengue incidence that typically follows monsoons in the Jaffna peninsula and elsewhere in Sri Lanka. It is possible however, that brackish water-developing Aedes constitute a perennial reservoir for DENV to maintain a basal level of dengue transmission in coastal areas of the peninsula during the dry season, and this supports increased transmission when monsoonal rains expand populations of fresh water Aedes.

Participation of 14-3-3ε and 14-3-3ζ proteins in the phagocytosis, component of cellular immune response, in Aedes mosquito cell lines.

BACKGROUND: Better knowledge of the innate immune system of insects will improve our understanding of mosquitoes as potential vectors of diverse pathogens. The ubiquitously expressed 14-3-3 protein family is evolutionarily conserved from yeast to mammals, and at least two isoforms of 14-3-3, the ε and ζ, have been identified in insects. These proteins have been shown to participate in both humoral and cellular immune responses in Drosophila. As mosquitoes of the genus Aedes are the primary vectors for arboviruses, causing several diseases such as dengue fever, yellow fever, Zika and chikungunya fevers, cell lines derived from these mosquitoes, Aag-2 from Aedes aegypti and C6/36 HT from Aedes albopictus, are currently used to study the insect immune system. Here, we investigated the role of 14-3-3 proteins (ε and ζ isoform) in phagocytosis, the main cellular immune responses executed by the insects, using Aedes spp. cell lines. RESULTS: We evaluated the mRNA and protein expression of 14-3-3ε and 14-3-3ζ in C6/36 HT and Aag-2 cells, and demonstrated that both proteins were localised in the cytoplasm. Further, in C6/36 HT cells treated with a 14-3-3 specific inhibitor we observed a notable modification of cell morphology with filopodia-like structure caused through cytoskeleton reorganisation (co-localization of 14-3-3 proteins with F-actin), more importantly the decrease in Salmonella typhimurium, Staphylococcus aureus and E. coli phagocytosis and reduction in phagolysosome formation. Additionally, silencing of 14-3-3ε and 14-3-3ζ expression by mean of specific DsiRNA confirmed the decreased phagocytosis and phagolysosome formation of pHrodo labelled E. coli and S. aureus bacteria by Aag-2 cells. CONCLUSION: The 14-3-3ε and 14-3-3ζ proteins modulate cytoskeletal remodelling, and are essential for phagocytosis of Gram-positive and Gram-negative bacteria in Aedes spp. cell lines.