Culex Pipiens Pallens Research Articles

RNAi-mediated knockdown of arginine kinase genes leads to high mortality and negatively affect reproduction and blood-feeding behavior of Culex pipiens pallens.

Arginine kinase (AK) is one of the crucial enzymes involved in energy metabolism in invertebrates, and has been proposed as the target for RNA interference (RNAi)-based control of agricultural insect pests. While there is only one AK gene in most insects, two AK genes were identified in Culex pipiens pallens, the primary vector of lymphatic filariasis and epidemic encephalitis. The full-length cDNA sequences of CpAK1 and CpAK2 genes were obtained by reverse transcription PCR(RT-PCR) and rapid amplification of cDNA ends (RACE). The expression levels of CpAK1 and CpAK2 in different developmental stages and tissues were detected by reverse transcription quantitative PCR (RT-qPCR). The role of CpAK1 and CpAK2 in the reproduction and blood feeding behavior was analyzed using RNA interference (RNAi). Full-length cDNAs of CpAK1 and CpAK2 were isolated from Cx. pipiens pallens. Analysis of the expression pattern revealed that the mRNA level of CpAK1 was significantly higher than CpAK2 in all development stages and tissues examined, and the expressions of both CpAK1 and CpAK2 were upregulated in response to blood feeding. The co-knockdown of CpAK1 and CpAK2 mediated by RNAi led to high mortality (74.3%) of adult female mosquitoes and decreased hatchability (59.9%). Remarkably, the blood feeding rate and the engorgement rate of the female mosquitoes were negatively affected by co-injection of dsRNAs targeting CpAK1 and CpAK2. CpAK1 and CpAK2 were detected in all developmental stages and tissues, but showed divergence in expression level. RNAi-mediated knockdown of AK genes leads to high mortality and negatively affect blood-feeding behavior of Cx. pipiens pallens, suggesting that AK could be used for the target of RNAi-based mosquito control in the future.

Carboxymethyl Chitosan-Modified Graphene Oxide as a Multifunctional Vector for Deltamethrin Delivery and pH-Responsive Controlled Release, Enhanced Leaf Affinity, and Improved Mosquito-Killing Activity.

Traditional deltamethrin (DM) formulations (e.g., emulsifiable concentrates, wettable powders, etc.) have significant disadvantages of poor water dispersion stability, burst release, weak leaf affinity, short duration, poor efficacy, and high environmental toxicity. A nanomaterial-based pesticide delivery system (PDS) has provided effective strategies for green preparation and synergism of pesticide formulations. In this article, we developed carboxymethyl chitosan (CMCS)-modified graphene oxide (GO) as a vector for DM and constructed a pH-responsive PDS for Culex pipiens pallens control. GO-CMCS possesses excellent pesticide loading performance for DM (loading rate 87.76%). After being loading on GO-CMCS, the GO-CMCS-DM has a significantly improved dispersion stability in water. The GO-CMCS-DM exhibits pH-responsive controlled release performance, which can sustain the release of DM into the medium, maintaining an effective long-term concentration. Additionally, the leaf adhesion of GO-CMCS-DM is better than that for free DM, which can improve the pesticide utilization. Therefore, GO-CMCS-DM has a prolonged persistent period and sustained activity against Culex pipiens pallens. Considering the industrialization potential of GO, we believe that GO will play an important role in the pest control and antiepidemic fields.

Chromosome-level assembly of Culex pipiens molestus and improved reference genome of Culex pipiens pallens (Culicidae, Diptera).

Culex pipiens molestus and Culex pipiens pallens are two distinct bioforms in the Culex pipiens complex that are important vectors of several pathogens and are widely distributed around the world. In the current study, we present a high-quality chromosome-level genome of Cx. pipiens f. molestus and describe the genetic characteristics of this genome. The assembly genome was 559.749 Mb with contig and scaffold N50 values of 200.952 Mb and 0.370 Mb, and more than 94.78% of the assembled bases were located on 3 chromosomes. A total of 19,399 protein-coding genes were predicted. Many gene families were expanded in the genome of Cx. pipiens f. molestus, particularly those of the chemosensory protein (CSP) and gustatory receptor (GR) gene families. In addition, utilizing Hi-C data, we improved the previously assembled draft genome of Cx. pipiens f. pallens, with scaffold N50 of 186.195 Mb and contig N50 of 0.749 Mb, and more than 97.02% of the assembled bases were located on three chromosomes. This reference genome provides a foundation for genome-based investigations of the unique ecological and evolutionary characteristics of Cx. pipiens f. molestus, and the findings in this study will help to elucidate the mechanisms involved in species divergence in the Culex pipiens complex.

Design, Synthesis, and Bioactivities of Novel Tryptophan Derivatives Containing 2,5-Diketopiperazine and Acyl Hydrazine Moieties.

Based on the scaffolds widely used in drug design, a series of novel tryptophan derivatives containing 2,5-diketopiperazine and acyl hydrazine moieties have been designed, synthesized, characterized, and evaluated for their biological activities. The bioassay results showed that the target compounds possessed moderate to good antiviral activities against tobacco mosaic virus (TMV), among which compounds 4, 9, 14, 19, and 24 showed higher inactivation, curative, and protection activities in vivo than that of ribavirin (39 ± 1, 37 ± 1, 39 ± 1 at 500 mg/L) and comparable to that of ningnanmycin (58 ± 1, 55 ± 1, 57 ± 1% at 500 mg/L). Thus, these compounds are a promising candidate for anti-TMV development. Most of these compounds showed broad-spectrum fungicidal activities against 13 kinds of phytopathogenic fungi and selective fungicidal activities against Alternaria solani, Phytophthora capsica, and Sclerotinia sclerotiorum. Additionally, some of these compounds exhibited larvicidal activities against Tetranychus cinnabarinus, Plutella xylostella, Culex pipiens pallens, Mythimna separata, Helicoverpa armigera, and Pyrausta nubilalis.

Detection and persistence of environmental DNA (eDNA) of the different developmental stages of a vector mosquito, Culex pipiens pallens.

Preventing mosquito-borne infectious diseases requires that vector mosquitoes are monitored and controlled. Targeting immature mosquitoes (eggs, larvae, and pupae), which have less mobility than adults, is an effective management approach. However, conducting these surveys is often difficult due to the limitations of morphological classification and survey costs. The application of environmental DNA (eDNA) analysis can solve these issues because it allows easy estimation of species distribution and morphology-independent species identification. Although a few previous studies have reported mosquito eDNA detection, there is a gap in knowledge regarding the dynamics related to the persistence of immature mosquito eDNA. We used Culex pipiens pallens, a vector of West Nile fever, as a model species. First, we developed a species-specific detection assay and confirmed its specificity using in silico and in vitro tests. Next, we conducted laboratory experiments using breeding tanks. Water samples were collected at each developmental stage. In addition, water samples were collected daily until the seventh day after emergence from the pupae. We quantified eDNA using real-time PCR with the developed assay to investigate the dynamics of mosquito eDNA. The specificity of the developed assay was confirmed by in silico and in vitro tests. Mosquito eDNA was detected at all developmental stages and detected up to seven days after emergence of pupae. In particular, high concentrations of eDNA were detected immediately after hatching from eggs and after emergence from pupae. Highly frequent positive eDNA signals were continuously detected between egg hatching and pupa hatching. Mosquito eDNA was detected immediately after the eggs were introduced, and eDNA-positive detections continued until pupae emergence, suggesting that eDNA analysis is useful for monitoring mosquito larvae. In the future, monitoring immature mosquitoes using eDNA analysis will contribute to prevent mosquito-borne infectious diseases.

Species diversity of the Culex pipiens complex in the Republic of Korea

AbstractThe Culex pipiens complex, a common species in urban areas, has not yet caused a disease in the Republic of Korea, although it is abundant across residential areas. This complex is a major vector of West Nile virus in the United States. Additionally, Culex pipiens molestus and Culex pipiens pallens in the Republic of Korea are known to have vector competence. However, it is difficult to morphologically distinguish Culex pipiens complex species in the Republic of Korea. In this study, samples were collected from 48 sites near residential areas in the Republic of Korea, and the distribution of Culex pipiens complex species was firstly analyzed using species‐specific primers. Culex p. molestus and Cx. p. pallens were collected in large numbers in all the study regions except Ulleungdo, where the number of samples collected was five. Of the total 1063 samples, 70.1% were Cx. p. molestus and 29.9% were Cx. p. pallens. As there has been no research on the diversity of these species in the Republic of Korea, the results of this study would provide important baseline data for further studies on the Culex pipiens complex.

Single amino acids set apparent temperature thresholds for heat-evoked activation of mosquito transient receptor potential channel TRPA1

Animals detect heat using thermosensitive transient receptor potential (TRP) channels. In insects, these include TRP ankyrin 1 (TRPA1), which in mosquitoes is crucial for noxious heat avoidance and thus is an appealing pest control target. However, the molecular basis for heat-evoked activation has not been fully elucidated, impeding both studies of the molecular evolution of temperature sensitivity and rational design of inhibitors. In TRPA1 and other thermosensitive TRPs, the N-terminal cytoplasmic ankyrin repeat (AR) domain has been suggested to participate in heat-evoked activation, but the lack of a structure containing the full AR domain has hindered our mechanistic understanding of its role. Here, we focused on elucidating the structural basis of apparent temperature threshold determination by taking advantage of two closely related mosquito TRPA1s from Aedes aegypti and Culex pipiens pallens with 86.9% protein sequence identity but a 10 °C difference in apparent temperature threshold. We identified two positions in the N-terminal cytoplasmic AR domain of these proteins, E417 (A. aegypti)/Q414 (C. pipiens) and R459 (A. aegypti)/Q456 (C. pipiens), at which a single exchange of amino acid identity was sufficient to change apparent thresholds by 5 to 7 °C. We further found that the role of these positions is conserved in TRPA1 of a third related species, Anopheles stephensi. Our results suggest a structural basis for temperature threshold determination as well as for the evolutionary adaptation of mosquito TRPA1 to the wide range of climates inhabited by mosquitoes.

Pipiserpin, a Culex factor Xa inhibitor, affects female reproductive capacity and serves as a potential target for mosquito control.

Mosquito control is still the main prevention and control measure for numerous mosquito-borne diseases causing millions of deaths each year. New strategies for mosquito control are in demand. Proteases play an important role in mosquito physiology, therefore this study explored the inhibition of a serpin (serine protease inhibitor) in mosquitoes and its effect on reproductive capacity. A factor Xa inhibitor homolog (named Pipiserpin) was amplified and identified in Culex pipiens pallens mosquitoes. We expressed a recombinant Pipiserpin protein in vitro against which a mouse antiserum was generated. We found that female mosquitoes expressed more Pipiserpin protein than male mosquitoes. After mating, female mosquitoes were fed with blood mixed with different amounts of antisera and results showed that consumption of Pipiserpin impeded ovary development and decreased eggs hatching rates compared to that of the pre-immune serum group. We identified a Culex mosquito factor Xa inhibitor, Pipiserpin, which affects female reproductive potential. Our results suggest that Pipiserpin may be a novel target for mosquito population control. The conclusions from our study on Cx. pipiens pallens might serve as a reference for the development of control measures for other mosquitoes as well. © 2022 Society of Chemical Industry.

CircRNA expression profiles in deltamethrin-susceptible and -resistant Culex pipiens pallens (Diptera: Culicidae)

The excessive and improper application of insecticides has caused the evolution of resistance in many mosquito populations, including Culex pipiens pallens (L.). Deltamethrin, a representative pyrethroid insecticide, is the most widely used synthetic insecticide in mosquito-borne control field. Comprehensively identifying genes and regulators associated with deltamethrin resistance and elucidating the manner in which they regulate this process is critical for effective control of mosquitoes. CircRNAs are the upstream regulatory factors of miRNAs and mRNAs, which play a role via the competitive endogenous RNA mechanism. In this study, we used high-throughput circRNA sequencing to identify circRNAs that were expressed differently in deltamethrin-susceptible strain (DS strain) and -resistant strain (DR strain) mosquitoes [NCBI Sequence Read Archive (SRA) database accession number: PRJNA714543]. We detected a total of 12,816 significantly differentially expressed circRNAs (DE-circRNAs). Among them, 6769 circRNAs were up-regulated and 6047 circRNAs were down-regulated in the DR strain compared to DS strain. Among the DE-circRNAs, we further screened that supercont3.352:252102|253283 was significantly over-expressed in the DR strain through qPCR multiple verification (P < 0.05).We used the divergent primer to amplify the rolling circle product and obtained the full-length sequence of supercont3.352:252102|253283 (GeneBank accession number: MW729338). Through software comparison and bioinformatics analysis, we predicted that supercont3.352:252102|253283 might participate in deltamethrin resistance by sponging cpp-miR-1671 and blocking its inhibition on CYP4G15.We further found that the expression of cpp-miR-1671 was significantly lower in DR strain (P < 0.01), while the expression of CYP4G15 was significantly higher in DR strain (P < 0.05).Taken together, the present study provided the most comprehensive circRNA expression profile of mosquitoes, and suggested that supercont3.352:252102|253283 might participate in deltamethrin resistance through the supercont3.352:252102|253283/cpp-miR-1671/CYP4G15 pathway.

CPR63 promotes pyrethroid resistance by increasing cuticle thickness in Culex pipiens pallens

The cuticle protein (CP) encoded by CPR63 plays a role in deltamethrin resistance in Culex pipiens pallens. Herein, we investigated the distribution of CPR63 transcripts in this organism and observed high expression levels in legs and wings. Furthermore, expression of CPR63 in the legs of deltamethrin-resistant (DR) strains was 2.17-fold higher than in deltamethrin-susceptible (DS) strains. Cuticle analysis of small interfering RNA (siRNA) groups by scanning electron microscopy (SEM) revealed a significantly thinner cuticle of the tarsi in the siCPR63 group than in the siNC (negative control siRNA) group. Transmission electron microscopy (TEM) revealed that the exocuticle and endocuticle thickness of the tarsi were significantly thinner, which contributes the thinner procuticle of tarsi in the siCPR63 group than in the siNC group. Our results suggested that CPR63 might contribute to the resistance phenotype by thickening the cuticle and thereby possibly increasing the tolerance of mosquitoes to deltamethrin.

Density and Distribution of the Mosquitoes Population Inhabiting Jeonbuk Region

Following the increase of vector-borne infectious diseases due to climate change and international exchange, the study collected mosquitoes using BL and BG twice a month for nine months, from March to November 2019, at cattle sheds, habitats for migratory birds, and city centers, after which they were classified, identified, and checked for the presence of flavivirus by using real time RT-PCR in order to classify and identify the species of disease vector mosquitoes in the Jeonbuk, estimate their population, and investigate their regional distribution and the presence of viral pathogen infection. A total of 8,234 mosquitoes were collected, which were categorized as seven genera and twelve species. In terms of environment types, the highest number of mosquitoes &#8211; 3,884 (Trap Index 215.8) &#8211; were collected at the Wanju cattle shed. In terms of mosquito populations, Anopheles spp. was the dominant species with 2,538 mosquitoes (30.9%) collected. As the dominant species, Anopheles spp. mosquitoes were first collected in April, and the greatest numbers were collected in the order of July, August, and June, with numbers gradually decreasing starting in August. In terms of the mosquito species collected in each environment, Anopheles spp. mosquitoes were collected in cattle sheds, Aedes vexans mosquitoes were collected in habitats for migratory birds, and Culex pipiens pallens mosquitoes were collected in city centers, revealing a difference in the species of mosquitoes collected depending on the environmental characteristics. The collected mosquitoes were tested for flavivirus using real time RT-PCR, but all tests turned out negative. The results of the collection confirmed the habitation of mosquitoes related to vector-borne diseases, predicting further results if an abnormal climate further progresses. The study is also expected to be used as comprehensive pest control standards for future mosquitoes.

Azobenzene-Avermectin B1a Derivatives for Optical Modulation of Insect Behaviors.

Naturally occurring avermectins are allosteric modulators of glutamate-gated chloride channels (GluCls) and possess exceptionally potent anthelmintic, acaricidal, and insecticidal activities. Here, we develop photoswitchable azobenzene-avermectin (ABAVM) derivatives, which can be photoactivated upon ultraviolet irradiation. After illumination, the best compound p-AB4″AVM had a 1.88-fold and 2.74-fold insecticidal activity enhancement toward Culex pipiens pallens and Mythimna separata larvae, respectively. p-AB4″AVM allows for optical regulation of dorsal unpaired median neuron membrane potential with a 2.15-fold fluorescence intensity decrease after illumination. p-AB4″AVM and p-AB5AVM enable optical modulation of the behavioral response of Culex pipiens pallens larvae with 1- and 4-fold reduced mobility upon irradiation, respectively. The ABAVMs could be used to reversibly manipulate GluCls with light and may be useful for the mechanistic study of macrocyclic lactone insecticides.

Inductions of a CYP6 cluster conferring deltamethrin resistance in colonized and field-collected Culex pipiens pallens.

Mosquitoes transmit many damaging vector-borne diseases. Unfortunately, the rise of insecticide resistance has become a major obstacle to mosquito control. A preliminary study showed that a CYP6 cluster is significant for deltamethrin resistance in colonized Culex pipiens pallens. Here, several field strains were collected to explore the association of the cluster in deltamethrin tolerance. We examined the effect of deltamethrin treatment on the cluster expression at a deltamethrin concentration of LC50 in these strains using five time points. As a result, both P450 induction and constitutive overexpression were associated with deltamethrin resistance. Deltamethrin could stimulate different expression sets in the P450 cluster in different strains, predominately correlated with the resistance level of the strain. Our results will offer more insight into working with the characterization of P450s related to insecticide resistance.

Surveillance of Culex mosquitoes in Jiangsu Province from 2018 to 2019

To investigate the distribution and density of Culex mosquito populations and the resistance of Culex pipiens pallens to insecticides in Jiangsu Province in 2018 and 2019. During the period from June to October in 2018 and 2019, six counties (districts, cities) were sampled in southern, northern and central Jiangsu Province as surveillance sites. The density of Culex mosquitoes was measured overnight using the light trapping technique. In addition, Culex pipiens pallens mosquitoes were collected from Hai'an of Nantong City and Yandu District of Yancheng City, central Jiangsu Province, and the sensitivity of female first filial generations to dichlorodiphenyltrichloroethane (DDT), malation, proposur, beta cypermethrin and deltamethrin was tested using the standard WHO insecticide susceptibility test assay. A total of 104 423 Culex mosquitoes were captured in six surveillance sites of Jiangsu Province in 2018 and 2019, and Culex quinquefasciatus (49.11%), Culex pipiens pallens (28.38%), and Culex tritaeniorhynchus (21.04%) were predominant species. The density of Culex mosquitoes started to increase since early June, peaked in July and tended to be low in late October. Culex pipiens pallens mosquitoes captured from Hai'an was susceptible to malation, while those from Yandu District were moderately resistant to malation. Culex pipiens pallens mosquitoes from both Yandu and Hai'an were moderately resistant to proposur, and were resistant to DDT, beta cypermethrin and deltamethrin. Culex quinquefasciatus, Culex pipiens pallens and Culex tritaeniorhynchus are predominant Culex species in Jiangsu Province. Culex pipiens pallens is resistant to DT, beta cypermethrin and deltamethrin in central Jiangsu Province.

MiR-279-3p regulates deltamethrin resistance through CYP325BB1 in Culex pipiens pallens

BackgroundThe overuse of insecticides to control insect vectors has promoted extensive insecticide resistance in mosquitoes. In this study, the functions of microRNA (miR)-279-3p and its target CYP325BB1 in the regulation of deltamethrin resistance in Culex pipiens pallens was investigated.MethodsQuantitative real-time reverse transcription PCR was used to detect the expression levels of miR-279-3p and CYP325BB1. Then, the dual-luciferase reporter assay system, RNA interference, CDC bottle bioassay and Cell Counting Kit-8 (CCK-8) assay were used to explore the roles of these molecules in deltamethrin resistance both in vivo and in vitro.ResultsThe expression patterns of miR-279-3p and CYP325BB1 were compared between deltamethrin-sensitive (DS-strain) and deltamethrin-resistant (DR-strain) mosquitoes. Luciferase activity was downregulated by miR-279-3p, the effect of which was ablated by a mutation of the putative binding site for CYP325BB1. In DR-strain mosquitoes, the expression of miR-279-3p was increased by microinjection and oral feeding of miR-279-3p agomir (mimic). CYP325BB1 mRNA levels were downregulated, which resulted in a higher mortality of the mosquitoes in miR-279-3p mimic-treated groups. In the DS-strain mosquitoes, microinjection of a miR-279-3p inhibitor decreased miR-279-3p expression, whereas the expression of CYP325BB1 was increased; the mortality of these mosquitoes decreased significantly. In addition, overexpression of pIB/V5-His-CYP325BB1 changed the sensitivity of C6/36 cells to deltamethrin in vitro. Also in DR-strain mosquitoes, downregulation of CYP325BB1 expression by microinjection of si-CYP325BB1 increased mosquito mortality in vivo.ConclusionsThese findings provide empirical evidence of the involvement of miRNAs in the regulation of insecticide resistance and indicate that miR-279-3p suppresses the expression of CYP325BB1, which in turn decreases deltamethrin resistance, resulting in increased mosquito mortality. Taken together, the results provide important information for use in the development of future mosquito control strategies.Graphical abstract

Microbiota and transcriptome changes of Culex pipiens pallens larvae exposed to Bacillus thuringiensis israelensis

Culex pipiens pallens is an important vector of lymphatic filariasis and epidemic encephalitis. Mosquito control is the main strategy used for the prevention of mosquito-borne diseases. Bacillus thuringiensis israelensis (Bti) is an entomopathogenic bacterium widely used in mosquito control. In this study, we profiled the microbiota and transcriptional response of the larvae of Cx. pipiens pallens exposed to different concentrations of Bti. The results demonstrated that Bti induced a significant effect on both the microbiota and gene expression of Cx. pipiens pallens. Compared to the control group, the predominant bacteria changed from Actinobacteria to Firmicutes, and with increase in the concentration of Bti, the abundance of Actinobacteria was gradually reduced. Similar changes were also detected at the genus level, where Bacillus replaced Microbacterium, becoming the predominant genus in Bti-exposed groups. Furthermore, alpha diversity analysis indicated that Bti exposure changed the diversity of the microbota, possibly because the dysbiosis caused by the Bti infection inhibits some bacteria and provides opportunities to other opportunistic taxa. Pathway analysis revealed significant enhancement for processes associated with sphingolipid metabolism, glutathione metabolism and glycerophospholipid metabolism between all Bti-exposed groups and control group. Additionally, genes associated with the Toll and Imd signaling pathway were found to be notably upregulated. Bti infection significantly changed the bacterial community of larvae of Cx. pipiens pallens.

Design, Synthesis, and Insecticidal Activity of Novel Triazone Derivatives Containing Sulfonamide or Sulfonimide Moieties.

On the basis of sulfonamide moieties widely used in drugs and the mode of action of pymetrozine and the structures of some transient receptor potential channel antagonists and to improve the insecticidal activities of triazone insecticides, a series of triazone derivatives containing sulfonamide or sulfonimide moieties were designed, synthesized, and characterized. The bioassay results showed that most of these derivatives containing sulfonamide moieties exhibited excellent insecticidal activities against Aphis craccivora. Especially, (substituted)phenylsulfonylamino triazones I-9 (LC50 = 2.9869 mg/kg), I-10 (LC50 = 3.4957 mg/kg), I-11 (LC50 = 2.3154 mg/kg), I-12 (LC50 = 4.1614 mg/kg), I-13 (LC50 = 2.1690 mg/kg), and I-14 (LC50 = 3.0077 mg/kg) exhibited much higher aphicidal activity than pymetrozine (LC50 = 6.0635 mg/kg). All monosulfonyl-substituted 4-amino-triazinones had better activities than the corresponding disulfonyl-substituted compounds. In addition, some derivatives also exhibited insecticidal activities against Culex pipiens pallens. Among Helicoverpa armigera, Mythimna separata, and Ostrinia nubilalis, compounds I-18 and I-21 exhibited good larvicidal activities (LC50 values were 0.6929 and 0.2592 mg/kg, respectively) against C. pipiens pallens.

Molecular detection and identification of Culex flavivirus in mosquito species from Jeju, Republic of Korea

BackgroundMosquito-borne flaviviruses are prime pathogens and have been a major hazard to humans and animals. They comprise several arthropod-borne viruses, including dengue virus, yellow fever virus, Japanese encephalitis virus, and West Nile virus. Culex flavivirus (CxFV) is a member of the insect-specific flavivirus (ISF) group belonging to the genus Flavivirus, which is widely distributed in a variety of mosquito populations.MethodsViral nucleic acid was extracted from adult mosquito pools and subjected to reverse transcriptase nested polymerase chain reaction (PCR) using target-specific primers for detecting CxFV nonstructural protein 5 (NS5). The PCR-positive samples were then sequenced, and a phylogenetic tree was constructed, including reference sequences obtained from GenBank.Results21 pools, belonging to Culex pipiens pallens (Cx. p. pallens) were found to be positive for the CxFV RNA sequence, with a minimum infection rate of 14.5/1000 mosquitoes. The phylogenetic analysis of the NS5 protein sequences indicated that the detected sequences were closely related to strains identified in China, with 95–98% sequence similarities.ConclusionOur findings highlight the presence of CxFV in Cx. p. pallens mosquito species in Jeju province, Republic of Korea. This is the first study reporting the prevalence of CxFV in Culex Pipiens (Cx. pipiens) host in the Jeju province, which can create possible interaction with other flaviviruses causing human and animal diseases. Although, mosquito-borne disease causing viruses were not identified properly, more detailed surveillance and investigation of both the host and viruses are essential to understand the prevalence, evolutionary relationship and genetic characteristic with other species.

Repellency of Veratraldehyde (3,4-Dimethoxy Benzaldehyde) against Mosquito Females and Tick Nymphs

Arthropod-borne infectious diseases cause many deaths and a major economic burden worldwide. Repellents play an important role in protecting people from infectious biting arthropods. The repellency of veratraldehyde, a known food additive, and the WJ-1041 formulation containing 10% veratraldehyde was tested against Aedes albopictus and Culex pipiens pallens females and Haemaphysalis longicornis nymphs using arm-in-cage, indoor or filter paper tests. Veratraldehyde exhibited repellency similar to or lower than that of n,n-diethyl-meta-toluamide (DEET) against A. albopictus, but in H. longicornis, the activity of veratraldehyde was better than that of DEET. The repellency of the 10% veratraldehyde solution was comparable to that of 20% DEET against the two mosquitoes. When comparing repellency between the WJ-1041 formulation (10% veratraldehyde) and 10% DEET against C. pipiens pallens, A. Albopictus and H. longicornis, the two showed similar repellency and complete protection time (CPT) values. However, there was a small difference depending on the tested insects. The absorption of veratraldehyde via skin was minimal, if at all. The pharmacokinetic parameters (Cmax and Tmax) of veratraldehyde in blood samples of rats were not different from those of the control group. Based on these results, veratraldehyde has high potential to be commercialized as a repellent agent against infectious disease-borne pests in the near future.

Physiological characterization of chitin synthase A responsible for the biosynthesis of cuticle chitin in Culex pipiens pallens (Diptera: Culicidae)

BackgroundThe pathogens transmitted by mosquitoes to humans and animals cause several emerging and resurgent infectious diseases. Increasing insecticide resistance requires rational action to control the target vector population. Chitin is indispensable for insect growth and development and absent from vertebrates and higher plants. Chitin synthase A (CHSA) is a crucial enzyme in chitin synthesis; therefore, identifying and characterizing how CHSA determines chitin content may contribute to the development of novel vector control strategies.ResultsThe injection of small interfering RNA targeting CHSA (siCHSA) to knockdown CHSA transcripts in larval, pupal and adult stages of Culex pipiens pallens resulted in the appearance of different lethal phenotypes. When larval and pupal stages were injected with siCHSA, CHSA knockdown prevented larval molting, pupation and adult eclosion, and affected the production of chitin and chitin degradation, which resulted in an ecdysis defect phenotype of mosquitoes. When siCHSA was injected into mosquitoes in the adult stage, CHSA knockdown also affected the laminar organization of the mesoderm and the formation of pseudo-orthogonal patterns of the large fibers of the endoderm.ConclusionWe provide a systematic and comprehensive description of the effects of CHSA on morphogenesis and metamorphosis. The results show that CHSA not only affects chitin synthesis during molting, but also might be involved in chitin degradation. Our results further show that CHSA is important for the structural integrity of the adult mosquito cuticle.Graphical