Transcriptome Analysis of Response to Zika Virus Infection in Two Aedes albopictus Strains with Different Vector Competence.

Zika virus (ZIKV), which is mainly transmitted by Aedes albopictus in temperate zones, can causes serious neurological disorders. However, the molecular mechanisms that influence the vector competence of Ae. albopictus for ZIKV are poorly understood. In this study, the vector competence of Ae. albopictus mosquitoes from Jinghong (JH) and Guangzhou (GZ) Cities of China were evaluated, and transcripts in the midgut and salivary gland tissues were sequenced on 10 days post-infection. The results showed that both Ae. albopictus JH and GZ strains were susceptible to ZIKV, but the GZ strain was more competent. The categories and functions of differentially expressed genes (DEGs) in response to ZIKV infection were quite different between tissues and strains. Through a bioinformatics analysis, a total of 59 DEGs that may affect vector competence were screened-among which, cytochrome P450 304a1 (CYP304a1) was the only gene significantly downregulated in both tissues of two strains. However, CYP304a1 did not influence ZIKV infection and replication in Ae. albopictus under the conditions set in this study. Our results demonstrated that the different vector competence of Ae. albopictus for ZIKV may be determined by the transcripts in the midgut and salivary gland, which will contribute to understanding ZIKV-mosquito interactions and develop arbovirus disease prevention strategies.

The polymorphism and geographical distribution of knockdown resistance of adult Anopheles sinensis populations in eastern China.

BACKGROUND: Anopheles sinensis is one of the major malaria vectors in China and other southeast Asian countries, including Vietnam, Cambodia, Thailand. Vector control is considered to be the critical measure for malaria control, while the increasing prevalence of insecticide resistance caused by long-term use of insecticides, especially pyrethroids, is threatening the successful control of An. sinensis. In order to understand the underlying resistance mechanisms involved and molecular basis, the principal malaria vector, An. sinensis from Jiangsu and Anhui provinces, Southeast China, was investigated. METHODS: The adult Anopheles mosquitoes were sampled from multiple sites across Jiangsu and Anhui provinces, and sufficient mosquitoes collected from eleven sites for insecticide susceptibility bioassays. The DIIS4-DIIS6 region of the para-type sodium channel gene was amplified and sequenced, then multiple PCR and Taqman assays were used to assess the frequencies of kdr mutations at the target gene. RESULTS: In the present study, most of the adult An. sinensis populations were pyrethroids resistant, which indicated the presence of kdr resistance mutations in the para-type sodium channel gene. Sequence analyses demonstrated the kdr mutation existed at codon 1014 in Jiangsu and Anhui provinces. In adult An. sinensis, three mutant types (TTT L1014F, TTC L1014F, and TGT L1014C) of kdr alleles were detected, while no wild type (TTG L1014) was observed. The TTC L1014F mutation was first reported in Anhui province. CONCLUSIONS: The highly polymorphic kdr alleles were observed in all the adult An. sinensis populations, which suggested that in-depth studies are required for carrying on insecticide resistance monitoring and specific resistance mechanisms studying into establish effective long-term malaria vector control program in eastern China.

Correction to: The polymorphism and geographical distribution of knockdown resistance of adult Anopheles sinensis populations in eastern China.

Please be advised that since publication of the original article [1] the authors have flagged that they omitted to provide the up-to-date version of Fig. 1 and, as such, the wrong version of Fig. 1 is present in the article.

Multiple-Insecticide Resistance and Classic Gene Mutations to Japanese Encephalitis Vector Culex tritaeniorhynchus from China.

Widespread resistance of insect pests to insecticides has been widely reported in China and there is consequently an urgent need to adjust pest management strategies appropriately. This requires detailed information on the extent and causes of resistance. The aim of the present study was to investigate levels of resistance to 5 insecticides among 12 strains of Culex tritaeniorhynchus, a major vector of Japanese encephalitis in China. Resistance to deltamethrin, beta-cypermethrin, permethrin, dichlorvos, and propoxur were measured using larval bioassays. The allelic frequency of knockdown resistance (kdr) and acetylcholinesterase (AChE) mutations were determined in all strains. Larval bioassay results indicated that the field strains collected from different sites were resistant to deltamethrin, beta-cypermethrin, permethrin, dichlorvos, and propoxur, with resistance ratio values ranging from 1.70- to 71.98-fold, 7.83- to 43.07-fold, 3.54- to 40.03-fold, 291.85- to 530.89-fold, and 51.32- to 108.83-fold, respectively. A polymerase chain reaction amplification of specific alleles method for individual was developed to detect genotypes of the AChE gene mutation F455W in Cx. tritaeniorhynchus. The frequency of the AChE gene mutation F455W was 100.00% in all strains, making this mutation of no value as a marker of resistance to organophosphorous and carbamate pesticides in Cx. tritaeniorhynchus in China. The kdr allele was present in all strains at frequencies of 10.00-29.55%. Regression analysis indicated a significant correlation between kdr allele frequencies and levels of resistance to deltamethrin, beta-cypermethrin, and permethrin. These results highlight the need to monitor and map insecticide resistance in Cx. tritaeniorhynchus and to adjust pesticide use to minimize the development of resistance in these mosquitoes.

An Unmanned Aerial Vehicle-Mounted Cold Mist Spray of Permethrin and Tetramethylfluthrin Targeting Aedes albopictus in China.

Aedes albopictus is the primary vector of dengue fever and dengue hemorrhagic fever in China. Although there are previous studies on the application of adulticides to control this species, the application methods have either been back-pack or vehicle-mounted systems. However, many sites are too large to be effectively treated with back-pack sprayers, and the lack of roads restricts the use of vehicle-mounted sprayers. This paper provides the first study of using unmanned aerial vehicles to conduct cold mist sprays on Ae. albopictus habitats. A spray containing 4% permethrin and 1% tetramethylfluthrin was applied at an effective application rate of 9.0 mg/m(2). This method reduced Ae. albopictus populations by more than 90%. The results indicate this novel spray system is a powerful method to achieve a rapid decline of mosquito population in Ae. albopictus habitats in China.

Occurrence and reassortment of avian influenza A (H7N9) viruses derived from coinfected birds in China.

UNLABELLED: Over the course of two waves of infection, H7N9 avian influenza A virus has caused 436 human infections and claimed 170 lives in China as of July 2014. To investigate the prevalence and genetic diversity of H7N9, we surveyed avian influenza viruses in poultry in Jiangsu province within the outbreak epicenter. We found frequent occurrence of H7N9/H9N2 coinfection in chickens. Molecular clock phylogenetic analysis confirms coinfection by H7N9/H9N2 viruses and also reveals that the identity of the H7N9 outbreak lineage is confounded by ongoing reassortment between outbreak viruses and diverse H9N2 viruses in domestic birds. Experimental inoculation of a coinfected sample in cell culture yielded two reassortant H7N9 strains with polymerase segments from the original H9N2 strain. Ongoing reassortment between the H7N9 outbreak lineage and diverse H9N2 viruses may generate new strains with the potential to infect humans, highlighting the need for continued viral surveillance in poultry and humans. IMPORTANCE: We found frequent occurrence of H7N9/H9N2 coinfection in chickens. The H7N9 outbreak lineage is confounded by ongoing reassortment between H7N9 and H9N2 viruses. The importance of H9N2 viruses as the source of novel avian influenza virus infections in humans requires continuous attention.

Artificial Selection for Different Host Preferences in Culex pipiens pallens (Diptera: Culicidae) Mosquitoes.

Most mosquito species display host preferences that are a crucial determinant of the transmission rate of mosquito-borne pathogens. Although a transgenic approach, based on driving genes for zoophily into vector populations, has been advocated as a malaria control strategy by the World Health Organization since 1982, the genes involved in mosquito host choice remain poorly understood. Culex pipiens pallens Coquillet mosquitoes were artificially selected for two different host preferences in a specially designed experimental enclosure. Of 3,035 mosquitoes obtained from larvae and pupae collected from the wild (the F0 generation), 27% preferentially fed on pigeons and 16% fed on mice. Following artificial selection for these host preferences over successive generations, the percentage of mosquitoes that preferred to feed on pigeons or mice gradually increased, eventually stabilizing at ∼55 and 34%, respectively, after the sixth generation. Intergenerational differences in host preferences were significant (P < 0.001). Furthermore, differences in host preferences between mosquitoes selected to prefer pigeons and those selected to prefer mice were both significant and consistent over almost six generations.

Host-Seeking Behavior and Arbovirus Detection in Mosquitoes of Habahe County, Xinjiang Uigur Autonomous Region, China.

Mosquitoes in Habahe County of Xinjiang Uigur Autonomous Region in China are considered a serious nuisance problem to local residents, but little is known of their role in enzootic disease. Therefore, host-seeking behavior and virus detection in mosquitoes were investigated in this study. Adult host-seeking mosquitoes were sampled using the Centers for Disease Control and Prevention (CDC) light traps operated at three locations in June through August 2008. Nine traps were used at each location at 3 different heights (1 m, 3 m, and 5 m). Seven mosquito species from 4 genera were collected by CDC light traps in different habitats. In total, 90,055 mosquitoes were captured, of which Aedes vexans was the most abundant species, comprising 88.02% of all mosquitoes collected. The second most abundant species was Anopheles messese, which comprised about 5.86%. Other species caught were Culex modestus (2.89%), Aedes caspius (1.11%), Coquillettidia richiardii (0.61%), Ae. dorsalis (1.36%), and An. hyrcanus (0.14%). About 93.5% of Ae. vexans individuals were caught in CO2-baited CDC light traps at 1 m above the ground. The highest numbers of Cx. modestus were caught at the highest trap level, 5 m above ground. Overall, significantly more mosquitoes of all species were collected at dusk than at dawn. Based on blood-meal analyses, Ae. vexans and An. messese fed on various vertebrate hosts, whereas Cx. modestus fed on ducks only. From a total of 335 mosquito pools tested, 10 pools of Ae. vexans were found positive for alphavirus. Comparison with the gene database revealed that the alphavirus deoxyribonucleic acid fragment obtained (GenBank accession no. HM160530) was 100% homologous at the nucleotide level to chikungunya virus isolate LK (PB) chik3408, chikungunya virus isolate SGEHICHD122508, and chikungunya virus strain FD080231. The results of this study suggest that ongoing, integrated mosquito and arbovirus surveillance is necessary in this river wetland.

Evaluation of Vegetation Treatment With Lambda-Cyhalothrin to Control Aedes vexans and Simulium maculatum in Northern China.

The mosquito Aedes vexans and black fly Simulium maculatum are important blood-sucking pests in northern China. Control of these pests using current methods is difficult because of complex larval developmental sites. In the present study, lambda-cyhalothrin was applied as a barrier spray to vegetation and various structures surrounding human dwellings for control against the adults of each species. The barrier spray resulted in an 88.2-99.4% reduction in Ae. vexans abundance and a 67.8-89.6% reduction in S. maculatum abundance for 2 wk postapplication. Our results indicated that lambda-cyhalothrin applied as a barrier spray to vegetation and other structures that serve as harborage for adults of both species could be adopted as part of an integrated pest management approach against these pests.

Host feeding patterns of mosquitoes in a rural malaria-endemic region in hainan island, china.

Malaria is endemic in Wangxia Village of Hainan Island. In this area little is known about the host seeking behavior and feeding habit of mosquitoes. Three sites representing the most common habitat types in the village were selected to study the host seeking behavior and feeding habit of mosquitoes. Of the total 9 species belonging to 4 genera (Armigeres, Culex, Aedes, and Anopheles) collected in Wangxia Village, Culex tritaeniorhynchus and Cx. pipiens quinquefasciatus were the most commonly collected species. Armigeres subalbatus and Anopheles sinensis were moderately common species. Blood meal analysis confirmed that Cx. tritaeniorhynchus and Cx. p. quinquefasciatus fed on multiple hosts, mainly poultry but occasionally other animals. Anopheles sinensis, a vector of malaria, fed predominately on cattle hosts, followed by humans. Anopheles maculatus and An. barbirostris fed on both humans and domestic animals. Our results indicate that most mosquitoes in this area preferred domestic animals over humans and showed a tendency to feed on multiple hosts within the same gonotrophic cycle. Therefore, the potential role of domestic animals in arbovirus transmission should be evaluated as part of a strategy for controlling mosquito-borne diseases in this region.

The effect of artificial light at night (ALAN) on the characteristics of diapause of Aedes albopictus.

BACKGROUND: Recently, the effect of artificial light at night (ALAN) on the physiology and behavior of insects has gradually attracted the attention of researchers and has become a new research topic. Aedes albopictus is an important vector that poses a great public health risk. Further studies on the diapause of Ae. albopictus can provide a basis for new vector control, and it is also worth exploring whether the effect of ALAN on the diapause of Ae. albopictus will provide a reference for the prevention and control of infectious diseases mediated by Ae. albopictus. METHODS: In this study, we experimentally studied the diapause characteristics of different geographical strains of Ae. albopictus under the interference of ALAN, explored the effect of ALAN on the diapause of Ae. albopictus and explored the molecular mechanism of ALAN on the diapause process through RNA-seq. RESULTS: As seen from the diapause incidence, Ae. albopictus of the same geographic strain showed a lower diapause incidence when exposed to ALAN. The differentially expressed genes (DEGs) were mainly enriched in signaling and metabolism-related pathways in the parental females and diapause eggs of the ALAN group. CONCLUSIONS: ALAN inhibits Ae. albopictus diapause. In the short photoperiod induced diapause of Ae. albopictus in temperate strain Beijing and subtropical strain Guangzhou, the disturbance of ALAN reduced the egg diapause rate and increased the egg hatching rate of Ae. albopictus, and the disturbance of ALAN also shortened the life cycle of Ae. albopictus eggs after hatching.

Replication and transmission of West Nile virus in simulated overwintering adults of Culex pipiens pallens (Diptera: Culicidae) in China.

West Nile virus is perhaps the widest distributed arbovirus globally, being maintained in nature by transmission among ornithophagic Culex mosquitoes and a wide-variety of birds. A factor contributing to the success of the WNV invasion is the ability of the virus to successfully overwinter. Experimental studies on the preservation and transmission of West Nile virus by Culex pipiens pallens under simulated overwintering conditions have shown that Cx. p. pallens has a preservative effect on West Nile virus under low-temperature conditions. After the temperature rises, the virus reproduces rapidly and can spread to susceptible animals while the mosquitos feed on blood. This research is of great significance for exploring how West Nile virus enters new areas and how the cycle of transmission is maintained China.

Population genetic characteristics of Aedes aegypti in 2019 and 2020 under the distinct circumstances of dengue outbreak and the COVID-19 pandemic in Yunnan Province, China.

Introduction: Since Aedes aegypti invaded Yunnan Province in 2002, its total population has continued to expand. Shi et al. used microsatellite and mitochondrial molecular markers to study the Ae. aegypti populations in Yunnan Province in 2015 and 2016, found that it showed high genetic diversity and genetic structure. However, there are few studies on the population genetic characteristics of Ae. aegypti in Yunnan Province under different levels of human intervention. This study mainly used two common types of molecular markers to analyze the genetic characteristics of Ae. aegypti, revealing the influence of different input, prevention and control pressures on the genetic diversity and structure of this species. Understanding the genetic characteristics of Ae. aegypti populations and clarifying the diversity, spread status, and source of invasion are essential for the prevention, control and elimination of this disease vector. Methods: We analyzed the genetic diversity and genetic structure of 22 populations sampled in Yunnan Province in 2019 and 17 populations sampled in 2020 through nine microsatellite loci and COI and ND4 fragments of mitochondrial DNA. In 2019, a total of 22 natural populations were obtained, each containing 30 samples, a total of 660 samples. In 2020, a total of 17 natural populations were obtained. Similarly, each population had 30 samples, and a total of 510 samples were obtained. Results: Analysis of Ae. aegypti populations in 2019 and 2020 based on microsatellite markers revealed 67 and 72 alleles, respectively. The average allelic richness of the populations in 2019 was 3.659, while that in 2020 was 3.965. The HWE analysis of the 22 populations sampled in 2019 revealed significant departure only in the QSH-2 population. The 17 populations sampled in 2020 were all in HWE. The average polymorphic information content (PIC) values were 0.546 and 0.545, respectively, showing high polymorphism. The average observed heterozygosity of the 2019 and 2020 populations was 0.538 and 0.514, respectively, and the expected average heterozygosity was 0.517 and 0.519, showing high genetic diversity in all mosquito populations. By analyzing the COI and ND4 fragments in the mitochondrial DNA of Ae. aegypti, the populations sampled in 2019 had a total of 10 COI haplotypes and 17 ND4 haplotypes. A total of 20 COI haplotypes were found in the populations sampled in 2020, and a total of 24 ND4 haplotypes were obtained. STRUCTURE, UPGMA and DAPC cluster analyses and a network diagram constructed based on COI and ND4 fragments showed that the populations of Ae. aegypti in Yunnan Province sampled in 2019 and 2020 could be divided into two clusters. At the beginning of 2020, due to the impact of COVID-19, the flow of goods between the port areas of Yunnan Province and neighboring countries was reduced, and the sterilization was more effective when goods enter the customs, leading to different immigration pressures on Ae. aegypti population in Yunnan Province between 2019 and 2020, the source populations of the 2019 and 2020 populations changed. Mantel test is generally used to detect the correlation between genetic distance and geographical distance, the analysis indicated that population geographic distance and genetic distance had a moderately significant correlation in 2019 and 2020 (2019: p < 0.05 R2 = 0.4807, 2020: p < 0.05 R2 = 0.4233). Conclusion: Ae. aegypti in Yunnan Province maintains a high degree of genetic diversity. Human interference is one reason for the changes in the genetic characteristics of this disease vector.

The predicted potential distribution of Aedes albopictus in China under the shared socioeconomic pathway (SSP)1-2.6.

Aedes albopictus (Skuse) (Diptera: Culicidae) is one of the 100 most invasive species in the world and represents a significant threat to public health. The distribution of Ae. albopictus has been expanding rapidly due to increased international trade, population movement, global warming and accelerated urbanization. Consequently, it is very important to know the potential distribution area of Ae. albopictus in advance for early warning and control of its spread and invasion. We randomly selected 282 distribution sites from 27 provincial-level administrative regions in China, and used the GARP and MaxEnt models to analyze and predict the current and future distribution areas of Ae. albopictus in China. The results showed that the current range of Ae. albopictus in China covers most provinces such as Yunnan and Guizhou Provinces, and the distribution of Ae. albopictus in border provinces such as Tibet, Gansu and Jilin Provinces tend to expand westwards. In addition, the potential distribution area of Ae. albopictus in China will continue to expand westwards due to future climate change under the SSP126 climate scenario. Furthermore, the results of environmental factor filtering showed that temperature and precipitation had a large effect on the distribution probability of Ae. albopictus.

Identification and quantitative analysis of genes encoding odorant binding proteins in Aedes albopictus (Diptera: Culicidae).

Odorant binding proteins (OBPs) play a critical role in mediating mosquito behaviors. In the current study, four AealOBP genes were cloned and sequenced. Basic Local Alignment Search Tool Program and phylogenetic analysis of the deduced amino acid sequences identified a unique putative ortholog in Aedes aegypti (L.) for each Aedes albopictus (Skuse) OBP. Quantitative analysis showed some AealOBPs with a strong female/male expression ratio, which we proposed to be involved in the host seeking of female mosquitoes. Other OBPs are expressed at higher levels in male antennae and are considered candidates for the detection of floral volatiles. The current study provides basis for further detailed molecular characterization of Ae. albopictus olfactory systems.

Survival and Replication of Zika Virus in Diapause Eggs of Aedes Albopictus From Beijing, China.

Zika virus (ZIKV) has emerged as a globally important arbovirus. The virus is primarily transmitted to humans through the bite of an infective Aedes albopictus in temperate area. Vertical transmission of ZIKV by Ae. albopictus is determined and has been suggested to be a means by which the virus could persist in nature. Ae. albopictus undergoes a well-characterized photoperiodic diapause. Viruses are harbored by overwintering mosquitoes in diapause that contributes to the resurgence of vertebrate diseases in the following spring, yet little is known about the impact of diapause on the regulation of viral replication and survival. The purpose of this study is to determine that Ae. albopictus in Beijing are highly susceptible to ZIKV (92.3%), and viable virus is passed to their organs of progeny via vertical transmission. Moreover, diapause eggs (diapause incidence 97.8%) had significantly lower minimum infection rates and filial infection rates of the first gonotrophic cycle than those of the second gonotrophic cycle in the short-day photoperiod group. Regarding the development of diapause eggs, the minimum infection rates and ZIKV RNA copy number increased significantly, suggesting that virus RNA replication occurred in the diapause eggs. Meanwhile, eggs from the ZIKV-infected mosquitoes had a significantly lower hatching rate compared with uninfected mosquitoes, implying an intriguing interaction between diapause eggs and virus. The findings here suggest that vertical transmission of ZIKV from diapause eggs to progeny may have a critical epidemiological role in the dissemination and maintenance of ZIKV circulating in the vector.

The Potential Vector Competence and Overwintering of West Nile Virus in Vector Aedes Albopictus in China.

West Nile virus (WNV) is an arbovirus, which causes widespread zoonotic disease globally. In China, it was first isolated in Jiashi County, Kashgar Region, Xinjiang in 2011. Determining the vector competence of WNV infection has important implications for the control of disease outbreaks. Four geographical strains of Aedes Albopictus (Ae. Albopictus) in China were allowed to feed on artificial infectious blood meal with WNV to determine the infection and transmission rate. The results indicated that four strains of Ae. Albopictus mosquitoes could infect and transmit WNV to 1- to 3-day-old Leghorn chickens. The infection rates of different strains were ranged from 16.7 to 60.0% and were statistically different (χ2 = 12.81, p < 0.05). The highest infection rate was obtained from the Shanghai strain (60.0%). The transmission rates of Ae. Albopictus Shanghai, Guangzhou, Beijing, and Chengdu strains were 28.6, 15.2, 13.3, and 6.7%, respectively. Furtherly, the results reveal that Ae. Albopictus Beijing strain infected orally can transmit WNV transovarially even the eggs are induced diapausing. The study confirmed that WNV could survive in the diapause eggs of Ae. Albopictus and could be transmitted to progeny after diapause termination. This is of great significance for clarifying that the WNV maintains its natural circulation in harsh environments through inter-epidemic seasons.

Wolbachia infection in field-collected Aedes aegypti in Yunnan Province, southwestern China.

Background: Wolbachia is gram-negative and common intracellular bacteria, which is maternally inherited endosymbionts and could expand their propagation in host populations by means of various manipulations. Recent reports reveal the natural infection of Wolbachia in Aedes Aegypti in Malaysia, India, Philippines, Thailand and the United States. At present, none of Wolbachia natural infection in Ae. aegypti has been reported in China. Methods: A total of 480 Ae. aegypti adult mosquitoes were collected from October and November 2018 based on the results of previous investigations and the distribution of Ae. aegypti in Yunnan. Each individual sample was processed and screened for the presence of Wolbachia by PCR with wsp primers. Phylogenetic trees for the wsp gene was constructed using the neighbour-joining method with 1,000 bootstrap replicates, and the p-distance distribution model of molecular evolution was applied. Results: 24 individual adult mosquito samples and 10 sample sites were positive for Wolbachia infection. The Wolbachia infection rate (IR) of each population ranged from 0 - 41.7%. The infection rate of group A alone was 0%-10%, the infection rate of group B alone was 0%-7.7%, and the infection rate of co-infection with A and B was 0-33.3%. Conclusions: Wolbachia infection in wild Ae. aegypti in China is the first report based on PCR amplification of the Wolbachia wsp gene. The Wolbachia infection is 5%, and the wAlbA and wAlbB strains were found to be prevalent in the natural population of Ae. aegypti in Yunnan Province.

Genetic Diversity and Population Genetic Structure of Aedes albopictus in the Yangtze River Basin, China.

Aedes albopictus is an indigenous primary vector of dengue and Zika viruses in China. Understanding the population spatial genetic structure, migration, and gene flow of vector species is critical to effectively preventing and controlling vector-borne diseases. The genetic variation and population structure of Ae. albopictus populations collected from 22 cities along the Yangtze River Basin were investigated with nine microsatellite loci and the mitochondrial CoxI gene. The polymorphic information content (PIC) values ranged from 0.534 to 0.871. The observed number of alleles (Na) values ranged from 5.455 to 11.455, and the effective number of alleles (Ne) values ranged from 3.106 to 4.041. The Shannon Index (I) ranged from 1.209 to 1.639. The observed heterozygosity (Ho) values ranged from 0.487 to 0.545. The FIS value ranged from 0.047 to 0.212. All Ae. albopictus populations were adequately allocated to three clades with significant genetic differences. Haplotype 2 is the most primitive molecular type and forms 26 other haplotypes after one or more site mutations. The rapid expansion of high-speed rail, aircraft routes and highways along the Yangtze River Basin have accelerated the dispersal and communication of mosquitoes, which appears to have contributed to inhibited population differentiation and promoted genetic diversity among Ae. albopictus populations.

Screening and analysis of immune-related genes of Aedes aegypti infected with DENV2.

Dengue virus type Ⅱ (DENV2) is a primary serotype responsible for the dengue fever epidemic, and Aedes aegypti is the main DENV2 vector. Understanding the Aedes aegypti immune mechanism against DENV2 is the basis for research on immune blockade in mosquitoes. Some preliminary studies lack validation in the literature, so this study was performed to further study and validate the potential target genes to provide a further basis for screening key target genes. We screened 51 genes possibly related to Aedes aegypti infection and immunity from the literature for further verification. First, bioinformatic methods such as GO, KEGG and PPI analysis were used, and then RT-qPCR was used to detect the changes in mRNA expression in the midguts and salivary glands of Aedes aegypti infected with DENV2.Bioinformatic analysis showed that mostly genes of the glucose metabolism pathway and myoprotein were influenced. In salivary glands, the Gst (xa) and Toll (xb) expression levels were significantly correlated with DENV2 load (y, lg[DENV2 RNA copies]), y = -3436xa+0.2287xb+3.8194 (adjusted R2 = 0.5563, F = 9.148, PF = 0.0045). In midguts, DENV2 load was significantly correlated with the relative Fba(R2 = 0.4381, t = 2.497, p < 0.05, df = 8), UcCr(R2 = 0.4072, t = 2.344, p < 0.05, df = 8) and Gbps1(R2 = 0.4678, t = 2.652, p < 0.05, df = 8) expression levels, but multiple regression did not yield significant results. This study shows that genes related to glucose metabolism and muscle proteins contribute to the interaction between Aedes aegypti and dengue virus. It was confirmed that SAAG-4, histone H4, endoplasmin, catalase and other genes are involved in the regulation of DENV2 infection in Aedes aegypti. It was revealed that GST and Toll in salivary glands may have antagonistic effects on the regulation of DENV2 load. Fba, UcCr and Gbps1 in the midgut may increase DENV2 load. These study results further condensed the potential target gene range of the Aedes aegypti immune mechanism against DENV2 infection and provided basic information for research on the Aedes aegypti in vivo blockade strategy against DENV2.