Suppressed expression of oxidoreductin-like protein, Oxidor, increases follicle degeneration and decreases survival during the overwintering diapause of the mosquito Culex pipiens.

Throughout diapause in mosquitoes, stress resistance and subsequent prolonged lifespan are a few important features of diapause that are crucial for overwintering success. In the mosquito Culex pipiens, we suggest that oxidoreductin-like protein is involved with these diapause characteristics for overwintering survival. Expression of oxidor was more than two-fold higher in early stage diapausing females compared to their non-diapausing counterparts. Suppression of the gene that encodes oxidoreductin-like protein by RNAi significantly increased the proportion of degenerating follicles in early-stage adult diapausing females. Inhibition of oxidor also significantly reduced the survivability of diapausing females which indicates that this protein plays a key role in protecting multiple tissues during early diapause.

First field evidence infection of Culex perexiguus by West Nile virus in Sahara Oasis of Algeria.

BACKGROUND & OBJECTIVES: West Nile virus (WNV) is considered one of the most widely distributed arboviruses in the world which is transmitted by several mosquito species including the Culex genus. Culex pipiens is the major vector of this virus in Europe and USA whereas in African countries, other species such as Cx. perexiguus is considered as an important vector. This paper aimed to study the mosquito species involved in WNV transmission in Aougrout, one of the highly populated Oasis of Timimoun Province in Algeria where human WNV neuroinvasive diseases are prevalent. METHODS: CDC light-traps were installed in animal and human shelters for three nights. Collected mosquitoes were pooled and real-time PCR was performed to detect and identify WNV lineages 1 and 2 in the samples. Results: CDC light-traps collected 270 mosquitoes belonging to three genera. Culex genus was predominant with Cx. perexiguus as main species followed by Aedes and Anopheles genus. A total of 33 pools were tested; one pool containing Cx. perexiguus was found positive for WNV lineage 1. INTERPRETATION & CONCLUSION: This study reports for the first time a WNV natural infection of Culex perexiguus in the study region indicating that species other than Cx. pipiens should be taken into consideration in WNV surveillance, especially in specific environments like Saharan Oasis ecosystem.

Experimental investigation of the susceptibility of Italian Culex pipiens mosquitoes to Zika virus infection.

We investigated the susceptibility of an Italian population of Culex pipiens mosquitoes to Zika virus (ZIKV) infection, tested in parallel with Aedes aegypti, as a positive control. We analysed mosquitoes at 0, 3, 7, 10, 14, 20 and 24 days after an infectious blood meal. Viral RNA was detected in the body of Cx. pipiens up to three days post-infection, but not at later time points. Our results indicate that Cx. pipiens is not susceptible to ZIKV infection.

The cuticle proteins: a putative role for deltamethrin resistance in Culex pipiens pallens.

Insecticide resistance has been a major public health challenge. It is impendent to study the mechanism on insecticide resistance. In our previous study, 14 differentially accumulated insect cuticle proteins (ICPs) based on insecticide resistance proteomes and transcriptomes were found in the deltamethrin-resistant (DR) and -susceptible (DS) strains of Culex pipiens pallens. To investigate if these ICPs are associated with deltamethrin resistance, different transcriptional levels of the 14 ICPs were detected in the DS and DR strains from laboratory and field populations by using quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of the 14 ICPs were also measured after short-term exposure of the DS strain to deltamethrin. The full-length complementary DNA (cDNA) of CpCPLCG5 gene, which encodes one of the 14 ICPs, was cloned from Cx. pipiens pallens. Homology analysis and phylogenetic analysis were carried out with some other insects. Furthermore, small interfering RNA (siRNA) was used to knockdown the expression level of CpCPLCG5 gene for characterizing its contribution to deltamethrin resistance. The results showed that the expression level of CpCPLCG5 gene was higher in DR strain than in DS strain both in laboratory and field populations while the other 13 ICPs were downregulated. The full-length cDNA of CpCPLCG5 gene was 732 bp, with the ORF of 390 bp and deduced 129 amino acids (GenBank/KF723314,2013). Knockdown of CpCPLCG5 gene increased the susceptibility of the DR strain while the expression level of the other 13 ICPs elevated. Our findings indicate that the cuticle proteins are associated with deltamethrin resistance in Cx. pipiens pallens.

Predicting Culex pipiens/restuans Population Dynamics Using a Weather-Driven Dynamic Compartmental Population Model.

Mosquitoes of the genus Culex are important vectors of a variety of arthropod-borne viral infections. In most of the northern parts of the USA, Cx. pipiens/restuans is the predominant representative of this genus. As vectors, they play a key role in the spreading of arboviruses and thus, knowledge of the population dynamic of mosquitoes is important to understand the disease ecology of these viruses. As poikilotherm animals, the vital rates of mosquitoes are highly dependent on ambient temperature, and also on precipitation. We present a compartmental model for the population dynamics of Cx. pipiens/restuans. The model is driven by temperature, precipitation, and daytime length (which can be calculated from the geographic latitude). For model evaluation, we used long-term mosquito capture data, which were averaged from multiple sites in Cook County, Illinois. The model fitted the observation data and was able to reproduce between-year differences in the abundance of the Cx. pipiens/restuans mosquitoes, as well as the different seasonal trends. Using this model, we evaluated the effectiveness of targeting different vital rates for mosquito control strategies. The final model is able to reproduce the weekly mean Cx. pipiens/restuans abundance for Cook County with a high accuracy, and over a long time period of 20 years.

Culex pipiens distribution in Tunisia: Identification of suitable areas through Random Forest and MaxEnt approaches.

BACKGROUND: Tunisia has experienced several West Nile virus (WNV) outbreaks since 1997. Yet, there is limited information on the spatial distribution of the main WNV mosquito vector Culex pipiens suitability at the national level. OBJECTIVES: In the present study, our aim was to predict and evaluate the potential and current distribution of Cx. pipiens in Tunisia. METHODS: To this end, two species distribution models were used, i.e. MaxEnt and Random Forest. Occurrence records for Cx. pipiens were obtained from adult and larvae sampled in Tunisia from 2014 to 2017. Climatic and human factors were used as predictors to model the Cx. pipiens geographical distribution. Mean decrease accuracy and mean decrease Gini indices were calculated to evaluate the importance of the impact of different environmental and human variables on the probability distribution of Cx. pipiens. RESULTS: Suitable habitats were mainly distributed next to oases, in the north and eastern part of the country. The most important predictor was the population density in both models. The study found out that the governorates of Monastir, Nabeul, Manouba, Ariana, Bizerte, Gabes, Medenine and Kairouan are at highest epidemic risk. CONCLUSIONS: The potential distribution of Cx. pipiens coincides geographically with the observed distribution of the disease in humans in Tunisia. Our study has the potential for driving control effort in the fight against West Nile vector in Tunisia.

Autochthonous Transmission of West Nile Virus by a New Vector in Iran, Vector-Host Interaction Modeling and Virulence Gene Determinants.

Using molecular techniques and bioinformatics tools, we studied the vector-host interactions and the molecular epidemiology of West Nile virus (WNV) in western Iran. Mosquitoes were collected during 2017 and 2018. DNA typing assays were used to study vector-host interactions. Mosquitoes were screened by RT-PCR for the genomes of five virus families. WNV-positive samples were fully sequenced and evolutionary tree and molecular architecture were constructed by Geneious software and SWISS-MODEL workspace, respectively. A total of 5028 mosquito specimens were collected and identified. The most prevalent species was Culex (Cx.) pipiens complex (57.3%). Analysis of the blood-feeding preferences of blood-fed mosquitoes revealed six mammalian and one bird species as hosts. One mosquito pool containing non-blood-fed Cx. theileri and one blood-fed Culex pipiens pipiens (Cpp.) biotype pipiens were positive for WNV. A phylogram indicated that the obtained WNV sequences belonged to lineage 2, subclade 2 g. Several amino acid substitutions suspected as virulence markers were observed in the Iranian WNV strains. The three-dimensional structural homology model of the E-protein identified hot spot domains known to facilitate virus invasion and neurotropism. The recent detection of WNV lineage 2 in mosquitoes from several regions of Iran in consecutive years suggests that the virus is established in the country.

The Effect of Weather Variables on Mosquito Activity: A Snapshot of the Main Point of Entry of Cyprus.

Mosquitoes are vectors of pathogens, causing human and animal diseases. Their ability to adapt and expand worldwide increases spread of mosquito-borne diseases. Climate changes contribute in enhancing these "epidemic conditions". Understanding the effect of weather variables on mosquito seasonality and host searching activity contributes towards risk control of the mosquito-borne disease outbreaks. To enable early detection of Aedes invasive species we developed a surveillance network for both invasive and native mosquitoes at the main point of entry for the first time in Cyprus. Mosquito sampling was carried out for one year (May 2017-June 2018), at bimonthly intervals around Limassol port. Morphological and molecular identification confirmed the presence of 5 species in the study region: Culex. pipiens, Aedes detritus, Ae. caspius, Culiseta longiareolata and Cs. annulata. No invasive Aedes mosquito species were detected. The Pearson's correlation and multiple linear regression were used to compare number of sampled mosquitoes and weather variables for three most numerous species (Cx. pipiens, Ae. detritus and Ae. caspius). The population densities of the most numerous species were highest from February to April. Number of Cx. pipiens (-0.48), Ae. detritus (-0.40) and Ae. caspius (-0.38) specimens sampled was negatively correlated with average daily temperature. Monthly relative humidity showed positive correlation with the numbers of the species sampled, Cx. pipiens (0.66) Ae. detritus (0.68), and Ae. caspius (0.71). Mosquito abundance of Cx. pipiens (0.97) and Ae. detritus (0.98) was strongly correlated to seasonal precipitation as well. Our work is a stepping stone to further stimulate implementation of International Health Regulations and implementation of early warning surveillance system for detection of invasive Aedes mosquitoes, native mosquitoes and arboviruses they may transmit. A network for the surveillance of both invasive and native mosquito species at the main point of entry for the first time in Cyprus was developed. Number of mosquitoes sampled was correlated with weather factors to identify parameters that might predict mosquito activity and species distribution to the prevention of international spread of vector mosquitoes and vector-borne diseases.

Bacillus thuringiensis novel toxin Epp is toxic to mosquitoes and prodenia litura larvae.

As a pathogenic bacterium, Bacillus thuringiensis (Bt) has become an alternative to chemical insecticides in commercial agricultural to control forestry pests and mosquitoes. To prevent pest resistance, many novel Bt strains have been isolated. Strain S3580-1 (WGS: VHPX0000000) used in this research and originally isolated from Hainan Qixianling National Forest Park (China) showed significant toxicity to Culex pipiens pallens. Here, using whole genome sequencing, assembly, and bioinformatics analysis, the predicted S3580-1CG_5163 (GenBank Accession No. MK124137) gene-encoded protein was found to share low homology with known toxins designated by the Bt toxin nomenclature system. It was considered to be an ETX/MTX2-type toxin and was designated Epp. Bioinformatics analysis showed that the predicted S3580-1CG_5163 gene-encoded protein Epp shared low identity with other known toxic protein sequences containing Cry-ETX/MTX conserved domains at the amino acid level, but significant similarity at the structural level. In addition, bioassays showed that Epp was toxic against Spodoptera litura (LC50 296.133 µg/mL; 95% FL 200.555-471.318 µg/mL) and Cx. pipiens pallens (LC50 322.193 µg/mL; 95% FL 238.217-477.243 µg/mL). On pathological observation, the peritrophic membrane of Cx. pipiens pallens larvae was degraded causing the midgut structure to become incomplete, resulting in larval death. Further bioassays are required to fully elucidate the insecticidal spectrum of the ETX/MTX2-type toxin Epp, and thereby provide future research directions.

Insecticide resistance of Culex pipiens (L.) populations (Diptera: Culicidae) from Riyadh city, Saudi Arabia: Status and overcome.

Three field populations of Cx. pipiens (L.) mosquitoes were collected from three different localities in Riyadh city. They were tested for developing resistance against commonly used insecticides to control mosquitoes in Riyadh. Two populations from Wadi Namar (WN1 and WN2) were highly resistant to deltamethrin (187.1- and 161.4-folds respectively). The field population from AL-Wadi district (AL-W) showed low resistance to lambda-cyhalothrin (3.8-folds) and moderate resistance to beta-cyfluthrin and bifenthrin (14- and 38.4-folds respectively). No resistance to fenitrothion was observed in WN1 population. Fenitrothion concentrations required to inhibit 50% of Acetylcholinesterase (AChE) activity in both WN1 population and the laboratory susceptible strain (S-LAB) were 0.073 and 0.078 ppm respectively. Piperonyl butoxide suppressed resistance to pyrethroid insecticides (>90%) in field populations indicating that oxidases and/or esterases play an important role in the reduction of pyrethroids toxicity. These results should be considered in the current mosquitoes control programs in Riyadh.