We show that, in the malaria vector Anopheles gambiae , expression of Cecropin 1 is regulated by REL2, an NF-κB-like transcription factor orthologous to Drosophila Relish. Through alternative splicing, REL2 produces a full-length (REL2-F) and a shorter (REL2-S) protein isoform lacking the inhibitory ankyrin repeats and death domain. RNA interference experiments show that, in contrast to Drosophila Relish, which responds solely to Gram-negative bacteria, the Anopheles REL2-F and REL2-S isoforms are involved in defense against the Gram-positive Staphylococcus aureus and the Gram-negative Escherichia coli bacteria, respectively. REL2-F also regulates the intensity of mosquito infection with the malaria parasite, Plasmodium berghei . The adaptor IMD shares the same activities as REL2-F. Microarray analysis identified 10 additional genes regulated by REL2, including CEC3 , GAM1 , and LRIM1 .

Aedes (Stegomyia) albopictus (Skuse) is an indigenous species and the predominant vector of dengue fever in China. Understanding of genetic diversity and structure of the mosquito would facilitate dengue prevention and vector control. Sympatric cryptic species have been identified in the Ae. albopictus subgroup in Southeast Asia; however, little is known about the presence and distribution of cryptic species in China. This study aimed to examine the genetic diversity, evaluate potential new cryptic sibling species, and assess the prevalence of Wolbachia infections in field populations. Aedes adult female specimens were collected from five provinces in southern and central China during 2015–2016. Morphological identification was performed under dissection microscope. The mitochondrial DNA cytochrome c oxidase subunit 1 (cox1, DNA barcoding) locus and the ribosomal DNA internal transcribed spacer region 2 (ITS2) marker were used to examine the genetic variation, evaluate cryptic sibling species, and population structure in the field populations. Screening for the presence of Wolbachia was performed using multiplex PCR. A total of 140 individual specimens with morphological characteristics similar to Ae. albopictus were sequenced for DNA barcoding. Among these, 129 specimens (92.1%) were confirmed and identified as Ae. albopictus. The remaining 11 specimens, from 2 provinces, were identified as 2 distinct sequence groups, which were confirmed by ITS2 marker sequencing, suggesting the existence of potential cryptic species of Ae. albopictus. In Ae. albopictus, we found significant genetic differentiation and population structure between populations collected from different climate zones. Medium to high frequencies of Wolbachia infections were observed in natural Ae. albopictus populations, whereas Wolbachia was infrequent or absent in cryptic species populations. Our findings highlight the population differentiation by climate zone and the presence of novel, cryptic Aedes species in China. The low prevalence of Wolbachia infections in cryptic species populations could reflect either a recent invasion of Wolbachia in Ae. albopictus or different host immune responses to this symbiont in the cryptic species. The study provides useful information for vector control and host-symbiont coevolution. Further study is needed to investigate the potential for arbovirus infection and disease transmission in the emerged cryptic species.

Background: Aedes albopictus is a highly invasive mosquito species and a major vector of numerous viral pathogens. Many recent dengue fever outbreaks in China have been caused solely by the vector. Mapping of the potential distribution ranges of Ae. albopictus is crucial for epidemic preparedness and the monitoring of vector populations for disease control. Climate is a key factor influencing the distribution of the species. Despite field studies indicating seasonal population variations, very little modeling work has been done to analyze how environmental conditions influence the seasonality of Ae. albopictus. The aim of the present study was to develop a model based on available observations, climatic and environmental data, and machine learning methods for the prediction of the potential seasonal ranges of Ae. albopictus in China. Methods: We collected comprehensive up-to-date surveillance data in China, particularly records from the northern distribution margin of Ae. albopictus. All records were assigned long-term (1970–2000) climatic data averages based on the WorldClim 2.0 data set. Machine learning regression tree models were developed using a 10-fold cross-validation method to predict the potential seasonal (or monthly) distribution ranges of Ae. albopictus in China at high resolution based on environmental conditions. The models were assessed based on sensitivity, specificity, and accuracy, using area under curve (AUC). WorldClim 2.0 and climatic and environmental data were used to produce environmental conduciveness (probability) prediction surfaces. Predicted probabilities were generated based on the averages of the 10 models. Results: During 1998–2017, Ae. albopictus was observed at 200 out of the 242 localities surveyed. In addition, at least 15 new Ae. albopictus occurrence sites lay outside the potential ranges that have been predicted using models previously. The average accuracy was 98.4% (97.1–99.5%), and the average AUC was 99.1% (95.6–99.9%). The predicted Ae. albopictus distribution in winter (December–February) was limited to a small subtropical-tropical area of China, and Ae. albopictus was predicted to occur in northern China only during the short summer season (usually June–September). The predicted distribution areas in summer could reach northeastern China bordering Russia and the eastern part of the Qinghai-Tibet Plateau in southwestern China. Ae. albopictus could remain active in expansive areas from central to southern China in October and November. Conclusions: Climate and environmental conditions are key factors influencing the seasonal distribution of Ae. albopictus in China. The areas predicted to potentially host Ae. albopictus seasonally in the present study could reach northeastern China and the eastern slope of the Qinghai-Tibet Plateau. Our results present new evidence and suggest the expansion of systematic vector population monitoring activities and regular re-assessment of epidemic risk potential.

Background: The Asian tiger mosquito,Aedes albopictus, is one of the 100 worst invasive species in the world and the vector for several arboviruses including dengue, Zika and chikungunya viruses. Understanding the population spatial genetic structure, migration, and gene flow of vector species is critical to effectively preventing and controlling vector-borne diseases. Little is known about the population structure and genetic differentiation of nativeAe. albopictusin China. The aim of this study was to examine the patterns of the spatial genetic structures of nativeAe. albopictuspopulations, and their relationship to dengue incidence, on a large geographical scale.Methods: During 2016–2018, adult femaleAe. albopictusmosquitoes were collected by human landing catch (HLC) or human-bait sweep-net collections in 34 localities across China. Thirteen microsatellite markers were used to examine the patterns of genetic diversity, population structure, and gene flow among nativeAe. albopictuspopulations. The correlation between population genetic indices and dengue incidence was also examined.Results: A total of 153 distinct alleles were identified at the 13 microsatellite loci in the tested populations. All loci were polymorphic, with the number of distinct alleles ranging from eight to sixteen. Genetic parameters such as PIC, heterozygosity, allelic richness and fixation index (F_ST) revealed highly polymorphic markers, high genetic diversity, and low population genetic differentiation. In addition, Bayesian analysis of population structure showed two distinct genetic groups in southern-western and eastern-central-northern China. The Mantel test indicated a positive correlation between genetic distance and geographical distance (R² = 0.245,P = 0.01). STRUCTURE analysis, PCoA and GLS interpolation analysis indicated thatAe. albopictuspopulations in China were regionally clustered. Gene flow and relatedness estimates were generally high between populations. We observed no correlation between population genetic indices of microsatellite loci inAe. albopictuspopulations and dengue incidence.Conclusion: Strong gene flow probably assisted by human activities inhibited population differentiation and promoted genetic diversity among populations ofAe. albopictus. This may represent a potential risk of rapid spread of mosquito-borne diseases. The spatial genetic structure, coupled with the association between genetic indices and dengue incidence, may have important implications for understanding the epidemiology, prevention, and control of vector-borne diseases.

Aedes albopictus is a vector of over 20 arboviruses that has spread throughout the world, mainly in the second half of the twentieth century. Approximately 50–100 million people are infected with dengue virus (DENV) transmitted by Aedes mosquitoes each year, leading to heavy economic burdens for both governments and individuals, among countless other negative consequences. Understanding the vector competence of vector species is critical for effectively preventing and controlling vector-borne diseases. Accordingly, in this study, vector competence was evaluated by quantitative analysis of DENV-2 loads in mosquito tissues (midguts, heads, and salivary glands) and whole mosquitoes through real-time quantitative polymerase chain reaction (RT-qPCR) analysis. Wolbachia and the expression of immune-associated genes (Rel1, Rel2, Dicer2, and STAT) in mosquitoes were also detected by RT-qPCR to explore their impact on vector competence. The amount of DENV-2 in the mosquito midguts, heads, and salivary glands from southern-western China were found to be lower than those from eastern-central-northern China. The DENV-2 loads in whole mosquitoes showed a negative correlation with Rel1 gene (r = -0.285, P = 0.011) and STAT gene expression levels (r = -0.289, P = 0.009). In terms of Wolbachia strains, the density of the wAlbB strain was found to be significantly higher than that of the wAlbA strain in the eight Ae. albopictus populations, and the relative density of the wAlbB strain in mosquitoes from southern-western China was higher than those from eastern-central-northern China. The relative density of the wAlbB strain showed a negative correlation with the mean loads of DENV-2 in the heads (r = -0.729, P = 0.040), salivary glands (r = -0.785, P = 0.021), and whole mosquitoes (r = -0.909, P = 0.002). Thus, there are lower DENV-2 loads in the mosquitoes from southern-western China, which may be related to the innate immunity of mosquitoes as affected by Rel1 in the Toll pathway, STAT in the JAK-STAT pathway, and the relative density of the wAlbB strain.

Aedes albopictus is a major vector for several tropical infectious diseases. Characterization of Ae. albopictus development under natural conditions is crucial for monitoring vector population expansion, dengue virus transmission, and disease outbreak preparedness. This study employed mosquito traits as a proxy to understanding life-table traits in mosquitoes using a semi–field study. Ae. albopictus larval and adult life-table experiments were conducted using microcosms under semi-field conditions in Guangzhou. Stage-specific development times and survivorship rates were determined and compared under semi-field conditions in different seasons from early summer (June) to winter (January), to determine the lower temperature limit for larval development and adult survivorship and reproductivity. The average egg- hatching rate was 60.1%, with the highest recorded in October (77.1%; mid-autumn). The larval development time was on average 13.2 days (range, 8.5–24.1 days), with the shortest time observed in September(8.7 days; early autumn) and longest in November (22.8 days). The pupation rates of Ae. albopictus larvae were on average 88.9% (range, 81.6–93.4%); they were stable from June to September but decreased from October to November. The adult emergence rates were on average 82.5% (range, 76.8–87.9%) and decreased from July to November. The median survival time of Ae. albopictus adults was on average 7.4 (range, 4.5–9.8), with the shortest time recorded in September. The average lifetime egg mass under semi-field conditions was 37.84 eggs/female. The larvae could develop into adults at temperatures as low as 12.3°C, and the adults could survive for 30.0 days at 16.3°C and still produce eggs. Overall, correlation analysis found that mean temperature and relative humidity were variables significantly affecting larval development and adult survivorship. Ae. albopictus larvae could develop and emerge and the adults could survive and produce eggs in early winter in Guangzhou. The major impact of changes in ambient temperature, relative humidity, and light intensity was on the egg hatching rates, adult survival time, and egg mass production, rather than on pupation or adult emergence rates.