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Abstract Background Culex tritaeniorhynchus is widely distributed in China, from Hainan Island in the south to Heilongjiang in the north, covering tropical, subtropical, and temperate climate zones. Culex tritaeniorhynchus carries 19 types of arboviruses. It is the main vector of the Japanese encephalitis virus (JEV), posing a serious threat to human health. Understanding the effects of environmental factors on Culex tritaeniorhynchus can provide important insights into its population structure or isolation patterns, which is currently unclear. Results In total, 138 COI haplotypes were detected in the 552 amplified sequences, and the haplotype diversity (Hd) value increased from temperate (0.534) to tropical (0.979) regions. The haplotype phylogeny analysis revealed that the haplotypes were divided into two high-support evolutionary branches. Temperate populations were predominantly distributed in evolutionary branch II, showing some genetic isolation from tropical/subtropical populations and less gene flow between groups. The neutral test results of HNQH (Qionghai) and HNHK(Haikou) populations were negative (P

Background: Culex tritaeniorhynchus is widely distributed in China, from Hainan Island in the south to Heilongjiang in the north, covering tropical, subtropical, and temperate climate zones. Culex tritaeniorhynchus carries 19 types of arboviruses. It is the main vector of the Japanese encephalitis virus (JEV), posing a serious threat to human health. Understanding the effects of environmental factors on Culex tritaeniorhynchus can provide important insights into its population structure or isolation patterns, which is currently unclear. Results: In total, 138 COI haplotypes were detected in the 552 amplified sequences, and the haplotype diversity (Hd) value increased from temperate (0.534) to tropical (0.979) regions. The haplotype phylogeny analysis revealed that the haplotypes were divided into two high-support evolutionary branches. Temperate populations were predominantly distributed in evolutionary branch II, showing some genetic isolation from tropical/subtropical populations and less gene flow between groups. The neutral test results of HNQH (Qionghai) and HNHK(Haikou) populations were negative (P < 0.05), indicating many low-frequency mutations in the populations and that the populations might be in the process of expansion. Moreover, Wolbachia infection was detected only in SDJN (Jining) (2.24%), and all Wolbachia genotypes belonged to supergroup B. To understand the influence of environmental factors on mosquito-borne viruses, we examined the prevalence of Culex tritaeniorhynchus infection in three ecological environments in Shandong Province. We discovered that the incidence of JEV infection was notably greater in Culex tritaeniorhynchus from lotus ponds compared to those from irrigation canal regions. In this study, the overall JEV infection rate was 15.27 per 1000, suggesting the current risk of Japanese encephalitis outbreaks in Shandong Province. Conclusions: Tropical and subtropical populations of Culex tritaeniorhynchus showed higher genetic diversity and those climatic conditions provide great advantages for the establishment and expansion of Culex tritaeniorhynchus. There are differences in JEV infection rates in wild populations of Culex tritaeniorhynchus under different ecological conditions. Our results suggest a complex interplay of genetic differentiation, population structure, and environmental factors in shaping the dynamics of Culex tritaeniorhynchus. The low prevalence of Wolbachia in wild populations may reflect the recent presence of Wolbachia invasion in Culex tritaeniorhynchus. [ABSTRACT FROM AUTHOR]

The Culex tritaeniorhynchus Giles, 1901 (Diptera: Culicidae) is major vector of Japanese encephalitis (JE) in China, and this study aimed to uncover the vector's spatiotemporal distribution and environmental correlation in Beijing. In study area, the Remote Sensing (RS), Global Position System (GPS), and Geographic Information System (GPS) were used to clarify the distribution characteristics of vector on spatial and temporal scales, and regressions analysis of cross-sectional study was performed to detect the environmental factors linked with the density and presence of Cx. tritaeniorhynchus. In study area, the scenic area was the major environmental area for breeding of the vector, August was the primary peak month, the new urban development area (NUDA) was major distribution subarea of Beijing, and the vector could be detected throughout the subarea of Beijing from June to September. In the scenic area, the total value of light index within buffer zones of 100 m (LT_100) and the total value of NDVI index within buffer zones of 800 m (NDVI_800) determined whether there was a positive or negative vector in the trapping sites, and the total value of NDVI index within buffer zones of 100 m (NDVI_100) and LT_100 was linked to the density of the vector. Our findings provide better insight into the spatio-temporal distribution pattern, associated environmental risk factors, and health risk of vector in Beijing. Based on the results here, we could predict the risk of JE and create and implement location-specific JE prevention and control measures to prevent future risks during the urbanization advancement of Beijing.

Information obtained about the blood feeding behavior of mosquito populations in nature is important for understanding their vectorial capacity and evaluating their potential role in the transmission of vector-borne diseases. This study investigated the feeding patterns of Culex pipiens, Anopheles sacharovi, and Cx. tritaeniorhynchus by using the ELISA method. A total of 435 blood-fed females were collected from the Aegean and Mediterranean regions of Türkiye between May 2021 and September 2023. The types and numbers of potential domestic animal hosts were recorded in each village. Of the 372 blood meals tested, 358 gave a single host positive result mainly on cow and human, chicken, respectively, in both villages. On the other hand, the host tendency for chicken was found in a very low degree in both villages. The human and other hosts were underselected, highlighting a marked zoophily for the vectors. Overall, these hosts were mainly selected in proportion to their abundance. These results highlight the zoophilic and exophagic behavior of these vectors. In conclusion, understanding the blood feeding behavior of mosquito populations is important for vector biology of these vectors. More research is needed to study the feeding behavior of outdoor mosquito populations and the influence of domestic animals on disease epidemiology. [ABSTRACT FROM AUTHOR]

Purpose: Mosquitoes are important vectors of pathogens that can affect humans and animals. Culex tritaeniorhynchus is an important vector of arboviruses such as Japanese encephalitis virus, West Nile virus among various human and animal communities. These diseases are of major public health concern and can have huge economic and health burdens in prevalent countries. Although populations of this important mosquito species have been detected in the Mediterranean and Aegean regions of Türkiye; little is known about its population structure. Our study is to examine the population genetics and genetic composition of Cx. tritaeniorhynchus mosquitoes collected from several localities using cytochrome oxidase subunit I (COI) and the NADH dehydrogenase subunit 5 genes (ND5). This is the first extensive study of Cx. tritaeniorhynchus in the mainland Türkiye with sampling spanning many of provinces. Methods: In this study, DNA extraction, amplification of mitochondrial COI and ND5 genes and population genetic analyses were performed on ten geographic populations of Culex tritaeniorhynchus in the Aegean and Mediterranean region of Türkiye. Results: Between 2019 and 2020, 96 samples were collected from 10 geographic populations in the Aegean and Mediterranean regions; they were molecularly analyzed and 139 sequences (50 sequence for COI and 89 sequence for ND5) were used to determine the population structure and genetic diversity. For ND5 gene region, the samples produced 24 haplotypes derived from 15 variable sites and for COI gene region, 43 haplotypes were derived from 17 variable sites. The haplotype for both gene regions was higher than nucleotide diversity. Haplotype phylogeny revealed two groups present in all populations. AMOVA test results show that the geographical populations were the same for all gene regions. Results suggest that Cx. tritaeniorhynchus is a native population in Türkiye, the species is progressing towards speciation and there is no genetic differentiation between provinces and regions. Conclusion: This study provides useful information on the molecular identifcation and genetic diversity of Cx. tritaeniorhynchus; these results are important to improve mosquito control programs. [ABSTRACT FROM AUTHOR]

Simple Summary: Mosquitoes, vectors of diseases like Japanese encephalitis, harbor complex microbial communities that influence their biology and disease transmission. The microbiome of Culex tritaeniorhynchus and Culex orientalis, key vectors of the Japanese encephalitis virus, remains poorly understood. Utilizing 16S rRNA Illumina sequencing, we investigated the microbiomes of these mosquitoes collected across diverse regions in the Republic of Korea. Our analysis revealed the presence of 36 prevalent bacterial families, with microbial composition variations predominantly influenced by geographic location. Moreover, the identification of species-specific biomarkers underscores the potential for ecological niche differentiation between Culex tritaeniorhynchus and Culex orientalis. This research sheds light on the intricate relationship between mosquito vectors, their microbiomes, and environmental factors, offering insights into vector ecology and disease transmission dynamics. Mosquitoes, the primary vectors of arboviruses, harbor a diverse microbiome that plays a crucial role in their development, immunity, and vector competence. The composition of the mosquito microbiome is heavily influenced by the environment and habitats. Therefore, identifying the relationship between the habitat and the mosquito's microbial community can improve the overall understanding of mosquito biology. However, The microbiome profiles of Culex tritaeniorhynchus and Culex orientalis, known as transmission vectors of the Japanese encephalitis virus, are poorly understood. Using 16S rRNA Illumina sequencing, we hereby investigated the microbial profiles in these two mosquito species collected in several areas in the Republic of Korea. Thirty-six prevalent bacterial families were identified from these mosquito species. The microbial composition variations were primarily influenced by the mosquito collecting sites. Moreover, species biomarkers were identified by utilizing the regional specificity of the mosquito microbiome. Based on the microbiome profiles representing high similarity, Culex orientalis may share an ecological niche with Culex tritaeniorhynchus. [ABSTRACT FROM AUTHOR]

Abstract Background Mosquitoes carry a variety of viruses that can cause disease in humans, animals and livestock. Surveys for viruses carried by wild mosquitoes can significantly contribute to surveillance efforts and early detection systems. In addition to mosquito-borne viruses, mosquitoes harbor many insect-specific viruses (ISVs). Quang Binh virus (QBV) is one such example, categorized as an ISV within the Flavivirus genus (family Flaviviridae). QBV has been specifically documented in Vietnam and China, with reports limited to several mosquito species. Methods The homogenate obtained from female mosquitoes was cultured on C6/36 (Aedes albopictus) and BHK-21 (baby hamster kidney) cell lines. Positive cultures were identified by reverse transcription-polymerase chain reaction (RT‒PCR) with taxon- or species-specific primers. Next-generation sequencing was employed to sequence the complete genomes of the identified positive samples. Subsequently, phylogenetic, gene homology, molecular evolutionary and genetic variation analyses were conducted. Result In 2021, a total of 32,177 adult female mosquitoes were collected from 15 counties in Guizhou Province, China. The predominant mosquito species identified were Culex tritaeniorhynchus, Armigeres subalbatus and Anopheles sinensis. Among the collected mosquitoes, three positive cultures were obtained from Cx. tritaeniorhynchus pools, revealing the presence of Quang Binh virus (QBV) RNA sequences. Phylogenetic analysis indicated that the three Guizhou isolates, along with the prototype isolate from Vietnam, formed distinct branches. These branches were primarily closely related to other QBV isolates reported in China. Comparative analysis revealed a high degree of nucleotide and amino acid homology between the Guizhou isolates and both Vietnamese and other indigenous Chinese isolates. Additionally, nonsynonymous single-nucleotide variants (SNVs) were observed in these strains compared to the QBV prototype strain. Conclusion This study represents the first report of QBV presences in Cx. tritaeniorhynchus mosquitoes in Guizhou Province, China. Phylogenetic tree analysis showed that the three Guizhou isolates were most closely related to the QBV genes found in China. In addition, the study of the genetic characteristics and variation of this virus provided a deeper understanding of QBV and enriched the baseline data of these insect-specific flaviviruses (ISFVs). Graphical Abstract

Wastes are the menace for the environment and weeds are one of the major categories of biowaste. They need to be managed properly for the well-being of environment. Sida acuta, also called as wireweed and Artemisia absinthium popularly known as wormwood, are the two major categories of weeds which are serious threat to the main vegetations and for animals as well, as they are toxic. Culex tritaeniorhynchus mosquito native to northern Asia is the vector of Japanese encephalitis, one of the concerned diseases declared by World Health Organization (WHO). Japanese encephalitis is endemic in 24 countries, causing 3 billion people at risk and has no fully developed cure. Therefore, the present investigation deals with integrated hand in hand management of weed waste and deadly disease vector with the utilization and blend of nanotechnology. In this study the larvicidal activity of the S. acuta and A. absinthium weeds silver nanoparticles were tested against the Japanese encephalitis vector. Green synthesis of silver nanoparticles was confirmed using Ultraviolet Visible (UV-Vis) Spectrophotometry, and peaks were observed at 439.2 nm with S. acuta and 477.6 nm with A. absinthium. The synthesized silver nanoparticles (AgNPs) were characterized using Field Emission-Scanning Electron Microscopy (FE-SEM), which depicted their spherical shape. The LC50 values for III instar larvae was found to be 0.121 μg/ml with S. acuta AgNPs and 0.104 μg/ml with AgNPs synthesized from A. absinthium. AgNPs of A. absinthium were found to be more effective larvicide in comparison to another weed taken into study. The study depicts a good strategy for hand in hand weed and deadly disease vector management. Nanolarvicides prepared using these weeds can be used in controlling Japanese encephalitis vector, as well as these can be explored for their larvicidal potential against vectors of other deadly mosquito borne diseases. [ABSTRACT FROM AUTHOR]

Background: Mosquitoes carry a variety of viruses that can cause disease in humans, animals and livestock. Surveys for viruses carried by wild mosquitoes can significantly contribute to surveillance efforts and early detection systems. In addition to mosquito-borne viruses, mosquitoes harbor many insect-specific viruses (ISVs). Quang Binh virus (QBV) is one such example, categorized as an ISV within the Flavivirus genus (family Flaviviridae). QBV has been specifically documented in Vietnam and China, with reports limited to several mosquito species. Methods: The homogenate obtained from female mosquitoes was cultured on C6/36 (Aedes albopictus) and BHK-21 (baby hamster kidney) cell lines. Positive cultures were identified by reverse transcription-polymerase chain reaction (RT‒PCR) with taxon- or species-specific primers. Next-generation sequencing was employed to sequence the complete genomes of the identified positive samples. Subsequently, phylogenetic, gene homology, molecular evolutionary and genetic variation analyses were conducted. Result: In 2021, a total of 32,177 adult female mosquitoes were collected from 15 counties in Guizhou Province, China. The predominant mosquito species identified were Culex tritaeniorhynchus, Armigeres subalbatus and Anopheles sinensis. Among the collected mosquitoes, three positive cultures were obtained from Cx. tritaeniorhynchus pools, revealing the presence of Quang Binh virus (QBV) RNA sequences. Phylogenetic analysis indicated that the three Guizhou isolates, along with the prototype isolate from Vietnam, formed distinct branches. These branches were primarily closely related to other QBV isolates reported in China. Comparative analysis revealed a high degree of nucleotide and amino acid homology between the Guizhou isolates and both Vietnamese and other indigenous Chinese isolates. Additionally, nonsynonymous single-nucleotide variants (SNVs) were observed in these strains compared to the QBV prototype strain. Conclusion: This study represents the first report of QBV presences in Cx. tritaeniorhynchus mosquitoes in Guizhou Province, China. Phylogenetic tree analysis showed that the three Guizhou isolates were most closely related to the QBV genes found in China. In addition, the study of the genetic characteristics and variation of this virus provided a deeper understanding of QBV and enriched the baseline data of these insect-specific flaviviruses (ISFVs). [ABSTRACT FROM AUTHOR]

Abstract Background The Getah virus (GETV) is a mosquito-borne Alphavirus (family Togaviridae) that is of significant importance in veterinary medicine. It has been associated with major polyarthritis outbreaks in animals, but there are insufficient data on its clinical symptoms in humans. Serological evidence of GETV exposure and the risk of zoonotic transmission makes GETV a potentially medically relevant arbovirus. However, minimal emphasis has been placed on investigating GETV vector transmission, which limits current knowledge of the factors facilitating the spread and outbreaks of GETV. Methods To examine the range of the mosquito hosts of GETV, we selected medically important mosquitoes, assessed them in vitro and in vivo and determined their relative competence in virus transmission. The susceptibility and growth kinetics of GETVs in various mosquito-derived cell lines were also determined and quantified using plaque assays. Vector competency assays were also conducted, and quantitative reverse transcription-PCR and plaque assays were used to determine the susceptibility and transmission capacity of each mosquito species evaluated in this study. Results GETV infection in all of the investigated mosquito cell lines resulted in detectable cytopathic effects. GETV reproduced the fastest in Culex tritaeniorhynchus- and Aedes albopictus-derived cell lines, as evidenced by the highest exponential titers we observed. Regarding viral RNA copy numbers, mosquito susceptibility to infection, spread, and transmission varied significantly between species. The highest vector competency indices for infection, dissemination and transmission were obtained for Cx. tritaeniorhynchus. This is the first study to investigate the ability of Ae. albopictus and Anopheles stephensi to transmit GETV, and the results emphasize the role and capacity of other mosquito species to transmit GETV upon exposure to GETV, in addition to the perceived vectors from which GETV has been isolated in nature. Conclusions This study highlights the importance of GETV vector competency studies to determine all possible transmission vectors, especially in endemic regions. Graphical Abstract

ABSTRACT Hubei mosquito virus 2 (HMV2) is a novel mosquito virus that was first identified in 2016 in Hubei Province, China. Until now, HMV2 has been shown to be endemic in some areas of China and Japan, but its biological characteristics, epidemiology, and pathogenicity are not yet known. This report describes the detection of HMV2 in mosquitoes that were collected in Shandong Province in 2019 and presents the first isolation and molecular characterization of the virus. In this study, a total of 2,813 mosquitoes were collected and then divided into 57 pools, according to location and species. qRT-PCR and nested PCR were performed to confirm the presence of HMV2, and its genomic features, phylogenetic relationships, growth characteristics, and potential pathogenicity were further analyzed. The results showed that HMV2 was detected in 28 of the 57 mosquito pools and that the minimum infection rate (MIR) for HMV2 was 1.00% (28/2,813). A HMV2 strain and 14 viral partial sequences were obtained from the HMV2-positive pools, including one complete genome sequence. A phylogenetic analysis revealed that HMV2 from Shandong Province shared over 90% identity with other reported isolates and was closely related to the Culex inatomii luteo-like virus. IMPORTANCE Our study provided important epidemiological evidence for the epidemic of HMV2 in Shandong Province. Here, we report the first isolation and molecular characteristics of this virus and enrich our knowledge of the distribution of HMV2 in China.

Mosquitoes, the primary vectors of arboviruses, harbor a diverse microbiome that plays a crucial role in their development, immunity, and vector competence. The composition of the mosquito microbiome is heavily influenced by the environment and habitats. Therefore, identifying the relationship between the habitat and the mosquito’s microbial community can improve the overall understanding of mosquito biology. However, The microbiome profiles of Culex tritaeniorhynchus and Culex orientalis, known as transmission vectors of the Japanese encephalitis virus, are poorly understood. Using 16S rRNA Illumina sequencing, we hereby investigated the microbial profiles in these two mosquito species collected in several areas in the Republic of Korea. Thirty-six prevalent bacterial families were identified from these mosquito species. The microbial composition variations were primarily influenced by the mosquito collecting sites. Moreover, species biomarkers were identified by utilizing the regional specificity of the mosquito microbiome. Based on the microbiome profiles representing high similarity, Culex orientalis may share an ecological niche with Culex tritaeniorhynchus.

Background: The Getah virus (GETV) is a mosquito-borne Alphavirus (family Togaviridae) that is of significant importance in veterinary medicine. It has been associated with major polyarthritis outbreaks in animals, but there are insufficient data on its clinical symptoms in humans. Serological evidence of GETV exposure and the risk of zoonotic transmission makes GETV a potentially medically relevant arbovirus. However, minimal emphasis has been placed on investigating GETV vector transmission, which limits current knowledge of the factors facilitating the spread and outbreaks of GETV. Methods: To examine the range of the mosquito hosts of GETV, we selected medically important mosquitoes, assessed them in vitro and in vivo and determined their relative competence in virus transmission. The susceptibility and growth kinetics of GETVs in various mosquito-derived cell lines were also determined and quantified using plaque assays. Vector competency assays were also conducted, and quantitative reverse transcription-PCR and plaque assays were used to determine the susceptibility and transmission capacity of each mosquito species evaluated in this study. Results: GETV infection in all of the investigated mosquito cell lines resulted in detectable cytopathic effects. GETV reproduced the fastest in Culex tritaeniorhynchus- and Aedes albopictus-derived cell lines, as evidenced by the highest exponential titers we observed. Regarding viral RNA copy numbers, mosquito susceptibility to infection, spread, and transmission varied significantly between species. The highest vector competency indices for infection, dissemination and transmission were obtained for Cx. tritaeniorhynchus. This is the first study to investigate the ability of Ae. albopictus and Anopheles stephensi to transmit GETV, and the results emphasize the role and capacity of other mosquito species to transmit GETV upon exposure to GETV, in addition to the perceived vectors from which GETV has been isolated in nature. Conclusions: This study highlights the importance of GETV vector competency studies to determine all possible transmission vectors, especially in endemic regions. [ABSTRACT FROM AUTHOR]

The Culex tritaeniorhynchus Giles, 1901 (Diptera: Culicidae) is major vector of Japanese encephalitis (JE) in China, and this study aimed to uncover the vector's spatiotemporal distribution and environmental correlation in Beijing. In study area, the Remote Sensing (RS), Global Position System (GPS), and Geographic Information System (GPS) were used to clarify the distribution characteristics of vector on spatial and temporal scales, and regressions analysis of cross-sectional study was performed to detect the environmental factors linked with the density and presence of Cx. tritaeniorhynchus . In study area, the scenic area was the major environmental area for breeding of the vector, August was the primary peak month, the new urban development area (NUDA) was major distribution subarea of Beijing, and the vector could be detected throughout the subarea of Beijing from June to September. In the scenic area, the total value of light index within buffer zones of 100 m (LT&#95;100) and the total value of NDVI index within buffer zones of 800 m (NDVI&#95;800) determined whether there was a positive or negative vector in the trapping sites, and the total value of NDVI index within buffer zones of 100 m (NDVI&#95;100) and LT&#95;100 was linked to the density of the vector. Our findings provide better insight into the spatio-temporal distribution pattern, associated environmental risk factors, and health risk of vector in Beijing. Based on the results here, we could predict the risk of JE and create and implement location-specific JE prevention and control measures to prevent future risks during the urbanization advancement of Beijing., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

To explore the Culex tritaeniorhynchuses-specific virome, 6400 C. tritaeniorhynchuses were collected in Honghe autonomous prefecture, China. Abundant virus sequences were obtained from 28 viral families using metavirome sequencing. Herein, several viruses in C. tritaeniorhynchuses virome were verified using the PCR technique, which covers Japanese encephalitis virus (JEV), Getah virus, and even Chikungunya virus (CHIKV). Seven JEV gene sequences were amplified successfully, of which JEV-China/CT2016E-1 shared the highest homology with the known JEV sequence isolated in Korea, 1946, with at least 96.1% nucleotide (nt) identity, which belonged to genotype III. Nine CHIKV gene sequences were amplified, which shared the highest with at least 93.0% nt identity with CHIKV from Thailand isolated in 2007, which was assigned to genotype Asian. Remarkably, CHIKV was isolated from C. tritaeniorhynchus in China for the first time. It was initially confirmed that the isolated virus CHIKV-China/CT2016-1 may increase infectivity after passaging in Vero cells from BHK-21 cells. Collectively, our study reveals the diversity, properties, and potential virus susceptibility dynamics of the C. tritaeniorhynchus virome and sheds new perspectives on the viral ecology in other important biological vectors. [ABSTRACT FROM AUTHOR]

This study investigated the feeding patterns of populations of Anopheles sacharovi, Culex tritaeniorhynchus, and Culex pipiens in the Mediterranean and Aegean regions of Turkey. Blood-fed females resting inside barns, houses, and chicken coops in the rural areas were collected between May 2017 and September 2019, and the genomic DNA from each female was isolated to determine the blood source after amplification of the mitochondrial cytochrome b gene region. Cytb results showed that out of 445 blood-fed An. sacharovi females 2 blood-fed from dogs, 9 from birds, and 434 from cows. The results also showed that all 216 blood-fed females Cx. tritaeniorhynchus captured fed from cows, 6 out of 97 Cx. pipiens females fed from horses and 91 from birds. The feeding pattern of mosquito populations can be affected by numerous factors such as sampling method, sample size, and host abundance. Although field studies on mosquito feeding patterns do not reveal the feeding preference of such vectors, they provide vital information to understanding of vector and host interactions in the research area. Moreover, these data can give an idea of possible human contact in an environment where various hosts are readily accessible to a given vector mosquito; this is important for the prevention of various human diseases caused by vector mosquitoes. [ABSTRACT FROM AUTHOR]

In order to study the spatial distribution feature of mosquito vectors (vectors of Japanese B Encephalitis (JBE) and malaria) in the village house of Guangxi province, China. The Yuhuachang village, one key mosquito-borne disease surveillance site for mosquito-borne diseases of Guangxi province, was selected as the study site. Mosquito vectors abundance was firstly assessed by Chinese manufactured light-traps in twenty-five village houses in the study site. Then, 3S(GPS-RS-GIS) analysis was performed to do the environmental factors quantitative Analysis. Finally, spatial statistical analysis and generalized linear regression were applied to analyze the relationship of the spatial distribution of mosquito vectors with environmental factors, using SPSS and Geoda (095i). As the results showed, the distribution of both vectors was not spatially auto-correlated. Additionally, the number of pigsty and pig was positively correlated with the density of mosquito vector of JBE, and the area of mid-season rice was positively correlated with density of malaria vector. Thus, this study not only provides a field basis for the prevention of mosquito-borne disease as well as its vector in the study city but also provides useful guidance for the warning aide on JBE and malaria prevention in Guangxi province, China. [ABSTRACT FROM AUTHOR]

Simple Summary: The disposal of agricultural waste products is an emerging concern and an alternative to this is the development of value-added products from these wastes. Here we extracted the essential oil from Citrus maxima (CMEO) and examined its larvicidal and pest control potentials. Results pointed out that CMEO can be effective biopesticides against two major insect pests of stored grains. Furthermore, CMEO had a significant larvicidal action against different mosquito species. This study provided useful information on the compositional aspects and insecticidal properties of CMEO. The wastes generated during the post-harvest handling of various agricultural commodities is rather under-utlilized. The peels of citrus fruits are often discarded as waste. Citrus peels are rich in essential oils and exhibit toxicity towards various insect species. The essential oils are also an eco-friendly option for insect pest management. The Citrus maxima peel essential oil (CMEO), a waste product, characterized it, and evaluated its potential for insect pest management. The major terpenoids present in CMEO are Limonene and α-Pinene. The CMEO displayed potentials in controlling the insect pests via contact and fumigant toxicity. Moreover, CMEO showed significant larvicidal activities against Culex tritaeniorhynchus and Aedes aegypti species of mosquitoes; however, Armigeres subalbatus was more resistant. The biological safety of the essential oil was also tested against the stored seeds, where no significant inhibition of seed germination was noticed compared to the control. Utilizing a waste product such as citrus peel for pest management can achieve the dual objective of waste utilization and eco-friendly pest management. Overall, the CMEO is therefore found to be a bioactive essential oil extracted from the wastes of pomelo (C. maxima). [ABSTRACT FROM AUTHOR]

Objective: To isolate the Japanese encephalitis virus carried by Culex tritaeniorhynchus in Dongchuan District of Yunnan Province and analyze its molecular characteristics, so as to provide insights into the prevention and control of Japanese encephalitis in Yunnan Province., Methods: Mosquito specimens were collected using mosquito-trapping lamps from pig farms in Batang Village and Xiaoxin Village, Dongchuan District, Kunming City, Yunnan Province in July 2016, and the mosquito species was identified according to the mosquito morphology. Then, 60 to 100 mosquitoes of each species served as a group and were ground. Baby hamster kidney-21 (BHK-21) cells and Aedes albopictus clone C6/36 cells were used for virus isolation, and positive isolates were identified using flavivirus primers. The positive isolates were amplified using reverse transcription polymerase chain reaction (RT-PCR) assay with 15 pairs of specific primers covering the full length of the genotype I Japanese encephalitis virus, and DNA sequence assembly was performed using the software SeqMan in the DNASTAR package. The obtained sequences were aligned with the complete sequences of 38 Japanese encephalitis virus downloaded from the GenBank with the software MegAlign, and the nucleotide and amino acid homology analyses of the obtained sequences were performed. The difference in amino acid sites was analyzed with the software GeneDoc, and phylogenetic trees were created based on the sequences of the coding region and E protein of the isolated Japanese encephalitis virus with the software Mega X. In addition, the secondary and tertiary structures of the E protein of the Japanese encephalitis virus were predicted using the online tool SOPMA and the software Swiss-Model., Results: A total of 5 820 mosquitoes were collected and 3 843 Cx. tritaeniorhynchus (66.03%) were identified according to the mosquito morphology. A positive virus isolate, termed YNDC55-33, was isolated from Cx. tritaeniorhynchoides following batches of virus isolation from mosquito specimens, and cytopathic effect was observed following inoculation into BHK-21 and C6/36 cells. The YNDC55-33 virus isolate was successfully amplified with the flavivirus primes, and a long sequence containing 300 nucleotides was obtained. Following sequence alignment using the BLAST tool, the sequence of the YNDC55-33 virus isolate had high homology with that of the genotype I Japanese encephalitis virus. A long sequence with 10 845 nucleotides in length, which encoded 3 432 amino acids, was obtained by splicing the full sequence of the YNDC55-33 virus isolate. Phylogenetic analysis based on the whole-genome sequence and E gene sequence of the YNDC55-33 virus isolate showed that the new YNDC55-33 virus isolate was most closely related to the genotype I Guizhou isolate (GenBank accession number: HM366552), with nucleotide homology of 98.5% and amino acid homology of 99.4%, and the YNDC55-33 virus isolate shared 97.96% ± 0.33% nucleotide homology and 99.35% ± 0.08% amino acid homology with other genotype I Japanese encephalitis virus isolates, and < 90% nucleotide homology and < 98% amino acid homology with other genotypes of Japanese encephalitis virus. The YNDC55-33 virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate differed at 16 amino acid sites on E gene, and 7 out of 8 key amino acid sites related to neurovirulence. The secondary and tertiary structures of the E protein of the YNDC55-33 virus isolate were predicted to be characterized by random coils., Conclusions: A genotype I Japanese encephalitis virus was isolated from Cx. tritaeniorhynchus in Dongchuan District, Kunming City. This virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate does not differ at antigenic epitopes-related key amino acid sites, and the major protein structure of the virus isolate is random coils. This study adds new data for the epidemiological distribution of Japanese encephalitis virus in Yunnan Province, which may provide insights into the prevention and control of Japanese encephalitis in the province.

• First report on the detection of LSDV in the saliva of mosquitoes. • LSDV was detected in the saliva of Ae. aegypti as early as 2 dpi. • LSDV remained detectable in the mosquitoes until 14 dpi. • EIP of Cx. tritaeniorhynchus and Cx. quinquefasciatus was 8 and 14 dpi respectively. • All three mosquito species may serve as biological vectors of LSDV. Lumpy skin disease virus (LSDV) is a transboundary viral disease in cattle and water buffaloes. Although this Poxvirus is supposedly transmitted by mechanical vectors, only a few studies have investigated the role of local vectors in the transmission of LSDV. This study examined the infection, dissemination, and transmission rates of LSDV in Aedes aegypti, Culex tritaeniorhynchus, and Culex quinquefasciatus following artificial membrane feeding of 102.7, 103.7, 104.7 TCID 50 /mL LSDV in sheep blood. The results demonstrated that these mosquito species were susceptible to LSDV, with Cx tritaeniorhynchus exhibiting significantly different characteristics from Ae. aegypti and Cx. quinquefasciatus. These three mosquito species were susceptible to LSDV. Ae. aegypti showed it as early as 2 days post-infection (dpi), indicating swift dissemination in this particular species. The extrinsic incubation period (EIP) of LSDV in Cx. tritaeniorhynchus and Cx. quinquefasciatus was 8 and 14 dpi, respectively. Ingestion of different viral titers in blood did not affect the infection, dissemination, or transmission rates of Cx. tritaeniorhynchus and Cx. quinquefasciatus. All rates remained consistently high at 8–14 dpi for Cx. tritaeniorhynchus. In all three species, LSDV remained detectable until 14 dpi. The present findings indicate that, Ae. aegypti, Cx. tritaeniorhynchus , and Cx. quinquefasciatus may act as vectors during the LSDV outbreak; their involvement may extend beyond being solely mechanical vectors. [Display omitted] [ABSTRACT FROM AUTHOR]

Recently, there has been a lot of interest in essential oils and plant-based extracts as potential bioactive ingredients. This study examined the antibacterial and antifungal effects of essential oils extracted from fresh peels of Citrus aurantifolia (lime), Citrus reticulata (mandarin), and Citrus paradisi (grapefruit) agsinst Escherichia coli (ATCC35218), Enterococcus faecalis (ATCC29212), Bacillus subtilis (ATCC-a-W), and Staphylococcus aureus (ATCC25923) and from Fusarium spp., Mucor spp., and Trichoderma viride. Additionally, the antilarvicidal activity of these oils was tested against Culex tritaeniorhynchus in a controlled laboratory environment. Using three distinct doses of 0.5 mg/ml, 1 mg/ml, and 2 mg/ml in the agar well-diffusion experiment, E. coli displayed a larger inhibition zone of 21 mm at the highest level of C. reticulata, but E. faecalis exhibited no inhibition at all for any of the three varieties of oil. The least amount of C. paradise oil used showed a minimal growth inhibition impact of around 5 mm on E. coli. T. viride was found to have larger inhibition zones (21.2 to 10 mm) for all citrus oils at the highest concentration. The fourth instar larval mortality rate was measured in each of the three replicates following a 24-hour treatment period. Following the observation period, no significant differences were seen in any of the three distinct concentrations that were evaluated. From all extracted oil, a maximum of 100% mortality was observed. According to the findings of these various studies, citrus oil has potential use as a medicine, an organic insecticide that can inhibit certain microorganisms and a low-cost, ecologically beneficial way of controlling mosquito larvae. Additionally, for possible future usage, screening of the variety of accessible phytochemicals contained in three distinct oils has to be investigated. [ABSTRACT FROM AUTHOR]

Hubei mosquito virus 2 (HMV2) is a novel mosquito virus that was first identified in 2016 in Hubei Province, China. Until now, HMV2 has been shown to be endemic in some areas of China and Japan, but its biological characteristics, epidemiology, and pathogenicity are not yet known. This report describes the detection of HMV2 in mosquitoes that were collected in Shandong Province in 2019 and presents the first isolation and molecular characterization of the virus. In this study, a total of 2,813 mosquitoes were collected and then divided into 57 pools, according to location and species. qRT-PCR and nested PCR were performed to confirm the presence of HMV2, and its genomic features, phylogenetic relationships, growth characteristics, and potential pathogenicity were further analyzed. The results showed that HMV2 was detected in 28 of the 57 mosquito pools and that the minimum infection rate (MIR) for HMV2 was 1.00% (28/2,813). A HMV2 strain and 14 viral partial sequences were obtained from the HMV2-positive pools, including one complete genome sequence. A phylogenetic analysis revealed that HMV2 from Shandong Province shared over 90% identity with other reported isolates and was closely related to the Culex inatomii luteo-like virus. IMPORTANCE Our study provided important epidemiological evidence for the epidemic of HMV2 in Shandong Province. Here, we report the first isolation and molecular characteristics of this virus and enrich our knowledge of the distribution of HMV2 in China., Competing Interests: The authors declare no conflict of interest.

In Asia, Culex mosquitoes are of particular interest because of their role in maintaining endemic mosquito-borne viral diseases, including the Japanese encephalitis virus (JEV). Nonetheless, host-feeding preferences, along with naturally infecting RNA viruses in certain Culex species, remain understudied. In this study, selected blood-fed mosquitoes were processed for avian and mammalian blood meal source identification. Concurrently, cell culture propagation and high-throughput sequencing (HTS) approaches were used to determine the RNA virome of Culex mosquitoes collected in Ishikawa Prefecture, Japan. The identification of blood meal sources from wild-caught Culex spp. revealed that Culex (Culex) tritaeniorhynchus Giles, 1901, has a robust preference toward wild boar (62%, 26/42), followed by heron (21%, 9/42). The other two species, Culex (Oculeomyia) bitaeniorhynchus Giles, 1901, and Culex (Culex) orientalis Edwards, 1921, showed a distinct preference for avian species, including migratory birds. From the HTS results, 34 virus sequences were detected, four of which were newly identified virus sequences of unclassified Aspiviridae, Qinviridae, Iflaviridae, and Picornaviridae. The absence of observable cytopathic effects in mammalian cells and phylogenetic analysis suggested that all identified virus sequences were insect-specific. Further investigations involving other mosquito populations collected in different areas are warranted to explore previously unknown vertebrate hosts that may be linked to JEV dispersal in nature., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Culex tritaeniorhynchus is the primary vector of Japanese encephalitis (JE) and has a wide global distribution. However, the current and future geographic distribution maps of Cx. tritaeniorhynchus in global are still incomplete. Our study aims to predict the potential distribution of Cx. tritaeniorhynchus in current and future conditions to provide a guideline for the formation and implementation of vector control strategies all over the world. We collected and screened the information on the occurrence of Cx. tritaeniorhynchus by searching the literature and online databases and used ten algorithms to investigate its global distribution and impact factors. Cx. tritaeniorhynchus had been detected in 41 countries from 5 continents. The final ensemble model (TSS = 0.864 and AUC = 0.982) indicated that human footprint was the most important factor for the occurrence of Cx. tritaeniorhynchus . The tropics and subtropics, including southeastern Asia, Central Africa, southeastern North America and eastern South America, showed high habitat suitability for Cx. tritaeniorhynchus . Cx. tritaeniorhynchus is predicted to have a wider distribution in all the continents, especially in Western Europe and South America in the future under two extreme emission scenarios (SSP5-8.5 and SSP1-2.6). Targeted strategies for the control and prevention of Cx. tritaeniorhynchus should be further strengthened.

Culex vishnui Theobald, 1901, a main vector of Japanese encephalitis virus (JEV), is widely distributed in the Oriental region where it often accounts for a great part of the culicid fauna. This species also has been found naturally infected with at least 13 other arboviruses of medical and veterinary importance. Females blood feed predominantly upon pigs and birds, but may readily bite cattle and humans. Because of its abundance, medical importance, and presence throughout ecological gradients among urban, peri-urban, and rural areas, Cx. vishnui potentially may serve as a bridge vector transmitting viruses from natural and wild hosts to humans. Being zoo- and anthropophagic, omnipresent in the Oriental region, and presenting strong resistance to many insecticide families, this overlooked mosquito species may pose a serious health risk in one of the most densely populated regions of the world., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

To explore the Culex tritaeniorhynchuses- specific virome, 6400 C . tritaeniorhynchuses were collected in Honghe autonomous prefecture, China. Abundant virus sequences were obtained from 28 viral families using metavirome sequencing. Herein, several viruses in C. tritaeniorhynchuses virome were verified using the PCR technique, which covers Japanese encephalitis virus (JEV), Getah virus, and even Chikungunya virus (CHIKV). Seven JEV gene sequences were amplified successfully, of which JEV-China/CT2016E-1 shared the highest homology with the known JEV sequence isolated in Korea, 1946, with at least 96.1% nucleotide (nt) identity, which belonged to genotype III. Nine CHIKV gene sequences were amplified, which shared the highest with at least 93.0% nt identity with CHIKV from Thailand isolated in 2007, which was assigned to genotype Asian. Remarkably, CHIKV was isolated from C. tritaeniorhynchus in China for the first time. It was initially confirmed that the isolated virus CHIKV-China/CT2016-1 may increase infectivity after passaging in Vero cells from BHK-21 cells. Collectively, our study reveals the diversity, properties, and potential virus susceptibility dynamics of the C. tritaeniorhynchus virome and sheds new perspectives on the viral ecology in other important biological vectors., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zhang, Peng, Li, Li, Zhang, Li and Xiao.)

Objectives: The aim of this study was to investigate the presence of Japanese encephalitis virus (JEV) in a rice-farming community in the Philippines and to determine its implications regarding the epidemiology of viral encephalitides in the Asia-Pacific Region., Methods: Mosquitoes were collected monthly from animal-baited traps close to flooded rice fields in two barangays (villages) in the Municipality of San Jose, Tarlac Province in Luzon, from May 2009 to July 2010. Virus was detected by nested reverse transcription PCR. Phylogenetic analysis of the amplified virus envelope gene was done using the maximum-likelihood method., Results: A total of 28 700 known vector mosquitoes were collected, namely Culex vishnui, Culex fuscocephala, Culex tritaeniorhynchus , and Culex gelidus . JEV genotype III was detected in C. tritaeniorhynchus , belonging to the same genotype but form a different clade from those reported in the 1980s and in 2020 in this country., Conclusions: Japanese encephalitis is associated with rice cultivation and the presence of infected mosquitoes in Tarlac, Philippines. It remains to be seen whether the observed genetic shift of genotype III to genotype I in Asia will in time have an impact on the epidemiology of Japanese encephalitis in the Philippines. For long-term disease control, regular surveillance and Japanese encephalitis immunization in children and travelers in high risk areas are recommended., (© 2022 The Authors.)