Pipiens Mosquitoes Research Articles

Unveiling Antimicrobial and Insecticidal Activities of Biosynthesized Selenium Nanoparticles Using Prickly Pear Peel Waste

In the current study, prickly pear peel waste (PPPW) extract was used for the biosynthesis of selenium nanoparticles through a green and eco-friendly method for the first time. The biosynthesized SeNPs were characterized using UV-Vis, XRD, FTIR, TEM, SEM, EDX, and mapping. Characterization results revealed that biosynthesized SeNPs were spherical, polydisperse, highly crystalline, and had sizes in the range of 10–87.4 nm. Antibacterial, antifungal, and insecticidal activities of biosynthesized SeNPs were evaluated. Results revealed that SeNPs exhibited promising antibacterial against Gram negative (E. coli and P. aeruginosa) and Gram positive (B. subtilis and S. aureus) bacteria where MICs were 125, 125, 62.5, and 15.62 µg/mL, respectively. Moreover, SeNPs showed potential antifungal activity toward Candida albicans and Cryptococcus neoformans where MICs were 3.9 and 7.81 µg/mL, respectively. Furthermore, tested crud extract and SeNPs severely induced larvicidal activity for tested mosquitoes with LC50 and LC90 of 219.841, 950.087 mg/L and 75.411, 208.289 mg/L, respectively. The fecundity and hatchability of C. pipiens mosquito were significantly decreased as applied concentrations increased either for the crude or the fabricated SeNPs extracts. In conclusion, the biosynthesized SeNPs using prickly pear peel waste have antibacterial, antifungal, and insecticidal activities, which can be used in biomedical and environmental applications.

Environmental Study of Effect of Some Plants Extracts on Culex pipiens mosquitoes

Background 
 In the nature there are many plants that play important roles to control mosquitoes, but some of these plants can increased their effect after mixed with another plant. For that, this study aimed to know the effect of mixture of two types of different plants on Culex pipiens mosquitoes in the larva stages and the process of hatching the eggs. 
 Materials and methods 
 In this research the effect of parsley water extract (Petroselinum crispum) and mosses water extract (Funaria sp.) have been taken as mixture in ratio (1:1) to know their effect on Culex pipiens mosquitoes after using different concentrations (4.5, 3.5, 2.5, 1.5, 0.5) mg/l from it.
 Results 
 The results show that percentage of mortalities for C. pipiens larva mosquitoes were (89, 72, 55, 42, 39)%, (89, 62, 56, 36, 38)%, (75, 67, 54, 46, 26)% and (77, 65, 63, 56, 40)% with LC50 (1.8, 2, 2.3, 1.3) mg/l for first, second, third and fourth stage respectively. While the percentage of hatching eggs that exposed to mixture extracts were (31, 43, 65, 71, 83)% with LC50 (3.2) mg/l. Results haven’t shown a significant differentiation among percentage of larva mortalities stages, but they have shown among percentage of hatching eggs in level of probability (0.05), also concentrations have a positive correlation coefficient with percentage of larva mortalities stages and negative with percentage of hatching eggs. 
 Conclusion
 The water extract mixture from water extract of parsley and mosses can be killed larva stages in low concentration and the mortality increases with increased concentration of extract, and can prevented eggs hatching.

Larvicidal and adulticidal activity of essential oils from plants of the Lamiaceae family against the West Nile virus vector, Culex pipiens (Diptera: Culicidae)

Culex pipiens mosquitoes are the most widely distributed primary vector of the West Nile virus worldwide. Many attempts for investigation of botanical pesticides to avoid the development of pesticide resistance to conventional synthetic pesticides that are recognized as a threat to the diversity of ecosystems. The study aimed to determine the components of three essential oils of Lamiaceae family, lavender (Lavandula angustifolia), peppermint (Mentha piperita L.), and rosemary (Rosmarinus officinalis L.) by gas chromatography-mass spectrometry (GC–MS) analysis. Furthermore, aimed to validate the insecticidal activities of these oils as larvicidal agents against the third instar larvae of Culex pipiens using five different concentrations (62.5, 125, 250, 500, and 1000 ppm) for each oil in five replicates and as an adulticidal agent against approximately three-day-old female adults of Cx. Pipiens using 0.5, 1, 2, 4, and 5% concentrations in three replicates. The results generally showed a dose-related response. At 1000 ppm, rosemary oil showed the highest larvicidal (100%) (LC50, 214.97 ppm), followed by peppermint oil (92.00% mortality and LC50 (269.35 ppm). Lavender oil showed the lowest efficacy with 87.20% mortality and LC50 (301.11 ppm). At 5% oil concentration, the highest knockdown rate at 1 h was recorded for lavender oil (95.55%), followed by peppermint oil (88.89%) and lastly rosemary oil (84.44%). After 24 h, rosemary oil showed the lowest adult mortality rate (88.89%; LC50, 1.44%), while lavender and peppermint oils both showed a 100% mortality rate, with (LC50, 0.81% and 0.91%, respectively). The chemical constituents of the oils consisted of monoterpenes and sesquiterpenes that determined their insecticidal activities against the target insect stage. The study proposed that rosemary essential oil may be useful for the control of Cx. pipiens larvae as part of an integrated water treatment strategy, and lavender and peppermint oils may be used in an integrated plan for adult’s control.

Cyclodepsipeptides: Isolation from Endophytic Fungi of Sarcophyton ehrenbergi and Verification of Their Larvicidal Activity via In-Vitro and In-Silico Studies.

Culex pipiens mosquitoes are vectors to many viruses and can transmit diseases such as filariasis and avian malaria. The present study evaluated the larvicidal activity of marine-derived endophytic fungi Aspergillus nomius and Aspergillus flavus from the soft coral Sarcophyton ehrenbergi along with two known cyclodepsipeptide compounds, scopularide A (1) and B (2), isolated from A. flavus extract, against third-instar larvae of C. pipiens, using distilled water as a negative control and toosenedanin as a positive control. The structures of the isolated compounds were confirmed by various spectroscopic analyses. The lethal concentrations (LC50 and LC90) were calculated by probit analysis. Scopularide A was the most potent after 96 h treatment, with LC50 and LC90 values of 58.96 and 994.31 ppm, respectively, and with 82.66% mortality at a concentration of 300 ppm. To unravel the biochemical mechanism of the tested extracts and compounds, their effects against protease, chitinase, phenoloxidases and lipase enzymes from the whole-body tissue of C. pipiens were evaluated after 72 h treatment at LC50 dose. Superior activity was observed for A. flavus extract against all tested enzymes. A molecular docking study was conducted for scopularide A and B on the four tested enzymes, to further verify the observed activity. Results revealed good binding affinities for both compounds as compared to the docked ligands, mainly via a number of hydrogen bonds. This was the first study to report the isolation of endophytic fungi A. flavus and A. nomius from the marine soft coral S. ehrenbergi. The endophytic fungal extract of A. flavus was found to be a promising source for a natural larvicidal agent against C. pipiens populations.

Evaluation efficiency of Metarhizium anisopliae fungi in larva mortality Culex pipiens

This study has targeted the evaluation of the efficiency metarhizium anisopliae fungi in different concentrations of bio control four stages Culex Pipiens mosquitoes. This study has been showed different results by different concentrations and exposure periods for all stages larva Culex Pipiens that was sensitive to selective concentrations 2x105, 2x107 and 2x109 spore/ml . The concentration 2x109 was recorded highest mortality in first stage reach (63.55), while lowest concentration While the rate of floating the same pours when concentration is 2x105 and reached (49.57) The rate of secondary and third, third and fourth phase has reached the top 2x109 (56.87, 48.55%, 44.80%), respectively, While the percharge rate itself is at least 2x105 (45.22%, 38.75%, 34.75%, 34.38%), respectively. With regard to exposure periods, the exposure period after (96) hours achieved a significant output at other exposure periods, where the rate of land phase The first (81.1%) is the rate of perhaps, while the exposure period (24) has given less rate for the same phase, amounting to 30.82%.

Insecticide Resistance in the Disease Vector Culex pipiens in Morocco: Intensity, Mechanisms, and Contribution in Insecticide Resistance Management

West Nile (WN) virus has been detected in Morocco in 1996, 2003, and 2010. One WN human case and 94 equine cases with 42 deaths were reported in the first outbreak. The second and the last epizootics were in Kénitra and Mohammedia regions, respectively. Culex pipiens mosquitoes were strongly suspected as WN vectors during these epizootics in Morocco. To help propose the first insecticide resistance management strategy in Morocco, we provide baseline data on the resistance status of C. pipiens mosquitoes to insecticides most used in public health in Morocco. We investigated the resistance intensity of C. pipiens and the role of monooxygenases in pyrethroids resistance using the synergist piperonyl butoxide (PBO) in three prefectures of central Morocco: Mohammedia, Benslimane, and Skhirate. Bioassays were carried out using WHO (World Health Organization) diagnostic test kits for adult mosquitoes, with DDT 4%; deltamethrin 0.05%, 0.25%, and 0.5%; bendiocarb 0.1%, 0.5%, and 1%; and malathion 5%. The synergist PBO 4% was used to check for the involvement of detoxification enzymes in pyrethroids resistant populations. Mosquito populations tested displayed high resistance intensity to pyrethroids and carbamates in Mohammedia and moderate resistance intensity in Skhirate. In Benslimane, the resistance intensity of C. pipiens was high to carbamate and moderate to pyrethroids. The pre-exposure to PBO restored full or partial susceptibility to pyrethroids in the different regions. According to our finding, Mohammedia and Benslimane are at risk level 3 of control failure, while Skhirate is at risk level 2 of it. The employment of IRM is therefore imperative in all study sites.

Concurrent Infection With Dengue and Chikungunya Viruses in Humans and Mosquitoes: A Field Survey in Lower Moshi, Tanzania

Introduction: Dengue and Chikungunya have re-emerged as important diseases of global concern. Co-infections with Dengue virus (DENV) and Chikungunya virus (CHIKV) could have serious outcomes if not diagnosed and managed optimally. However, the key focal points for the maintenance of CHIKV and DENV infections and the extent of their co-infection remain poorly understood in many geo-ecologically distinct parts of Tanzania. Objective: We aimed to comparatively examine the prevalence and factors for seropositivity to DENV and CHIKV and their infection rates in humans and mosquitoes Methods: A cross-sectional study was performed in the Lower Moshi area of the Kilimanjaro region from April to July 2020. DENV and CHIKV exposure was determined by detecting IgM to the viruses using enzyme linked immunosorbent assay whereas infection was determined by real time quantitative polymerase chain reaction (RT-qPCR) assay. Results: Insecticide Treated Bed Net (ITN) use (χ2=3.504; p< 0.05), being ≥7 individuals living in the same household (χ2=4.655; p<0.05) and a recent travel to an urban destination (χ2=3.39; p< 0.05) were the only factors associated with CHIKV seropositivity. ITN use was the only factor associated with CHIKV infection (χ2=5.204; p<0.05). A recent travel to an urban destination (χ2=4.401; p< 0.05) was the only factor associated with DENV seropositivity. Five (1.5%) Ae. aegypti pools were positive for CHIKV whereas 1 (0.3%) was positive for DENV. Two Cx. pipiens, pools (1.9%) were positive for CHIKV. None of the Cx. pipiens mosquitoes was positive for DENV. No associations between DENV and CHIKV seropositivity was observed in humans but DENV infection was strongly associated with CHIKV infection (χ2 = 238.45; p<0.01). CHIKV infection was observed to be consistently higher in both, humans and mosquitoes. Conclusion: Detection of DENV and CHIKV in both humans and vector mosquitoes confirms that both viruses are actively circulating in the Lower Moshi area of Kilimanjaro region in Tanzania. Our findings point out the Lower Moshi area as a potential focal point for the maintenance of the two viruses and possibly other vector borne viruses. We call upon sustained active surveillance of arboviruses and other re-emerging infections to be better prepared for possible outbreaks by the viruses.

Modelling the temperature suitability for the risk of West Nile Virus establishment in European Culex pipiens populations.

Increases in temperature and extreme weather events due to global warming can create an environment that is beneficial to mosquito populations, changing and possibly increasing the suitable geographical range for many vector-borne diseases. West Nile Virus (WNV) is a flavivirus, maintained in a mosquito-avian host cycle that is usually asymptomatic but can cause primarily flu-like symptoms in human and equid accidental hosts. In rare circumstances, serious disease and death are possible outcomes for both humans and horses. The main European vector of WNV is the Culex pipiens mosquito. This study examines the effect of environmental temperature on WNV establishment in Europe via Culex pipiens populations through use of a basic reproduction number ( ) model. A metric of thermal suitability derived from was developed by collating thermal responses of different Culex pipiens traits and combining them through use of a next-generation matrix. WNV establishment was determined to be possible between 14°C and 34.3°C, with the optimal temperature at 23.7°C. The suitability measure was plotted against monthly average temperatures in 2020 and the number of months with high suitability mapped across Europe. The average number of suitable months for each year from 2013 to 2019 was also calculated and validated with reported equine West Nile fever cases from 2013 to 2019. The widespread thermal suitability for WNV establishment highlights the importance of European surveillance for this disease and the need for increased research into mosquito and bird distribution.

Detection and molecular characterization of West Nile virus in Culex pipiens mosquitoes in Central Macedonia, Greece, 2019–2021

Since 2010 when West Nile virus (WNV) emerged in Greece, it causes seasonal outbreaks of human infections almost every year. During May-October of 2019–2021 a total number of 51,504 Culex pipiens mosquitoes were trapped in all seven regional units of Central Macedonia in northern Greece. They were grouped into 1099 pools and tested for WNV. The virus was detected in 5% of the mosquito pools (1.5%, 3.6% and 9.6% pools in 2019, 2020, and 2021, respectively), with significant rate differences among the regional units and years. The highest maximum likelihood estimation for WNV infection rates calculated per 1000 mosquitoes for 2019 and 2020 were 1.89 and 3.84 in Serres, and 7.08 for 2021 in Pella regional unit. Sixteen whole genome sequences were taken by applying a recently described PCR-based next generation sequencing protocol. Phylogenetic analysis showed that the sequences belonged to the Central European clade of WNV lineage 2, and that a virus strain introduced in Greece in 2019 continued to circulate and spread further during 2020–2021. The data are useful for public health and mosquito control programs' operational scheduling, while the whole genome sequences are an added value for molecular epidemiology and evolutionary studies.

Comparison of Mosquito Larvicidal Formulations of Diflubenzuron on Culex Pipiens Mosquitoes in Belgrade, Serbia

Abstract Culex mosquitos are important infectious diseases vectors in temperate and tropical regions of the World, affecting nearly 350 million people in both developed and developing countries. Our approach was to “recycle” the well-established larvicide, and by studying the tablets, pellets and granules as floating or sinking formulations, we found a method to optimise the use of diflubenzuron against Culex pipiens mosquitoes in field conditions. A standard WHO procedure was used to test the larvicide efficacy. The combined efficacy of all floating formulations was 10.7% higher than sinking preparations (p-value =0.002) and that maximised throughout the experiment on days 14 and 21, (p-values 0.012 and 0.008, respectively). All floating formulations kept their efficacies above 70% until day 21 of the experiment, while the mortality of sinking designs dropped significantly after day 14. The lowest efficacy was observed when sinking granules were used and the highest when floating tablets were applied in the canals. Only the floating tablets showed no significant changes in efficacy from day 1 to 21, but then that efficacy drops sharp until day 35. Since the larvae spend most of their time on the surface of the slow-moving waters to breathe, and floating pellets and tablets are made of materials that can serve as food sources, we conclude that these formulations have a higher efficacy, at least in conditions of non- or slow-moving waters. This study shows the importance of a systematic approach to reformulation of old, already proven and environmentally safe larvicides which can control the mosquito populations and their spreading of various pathogens.

Effect of peppermint and anise oil and pesticide SIVANTO in the control of Culex pipiens (Diptera:Culicidae) mosquitoes

The study dealt with the effect of peppermint oil and anise oil and the pesticide SIVANTO in the control of C. pipiens mosquitoes, and the concentrations used for peppermint oil and anise oil and pesticide SIVANTO (0.5,1,1.5,2) the lowest killing rate for peppermint oil was 2.6% after 48 hours of treatment, and the highest killing rate was 4.6% after 48 hours Of the treatment, while anise oil had the lowest killing rate of 3.3% after 48 hours of treatment, and the highest killing rate of 4.6% after 48 hours of treatment, and the lowest killing rate of SIVANTO was 8% after 48 hours of treatment, and the highest killing rate of 10% after treatment. 48 hours of treatment compared to the control treatment in which no homicide occurred.

Impacts of fungal entomopathogens on survival and immune responses of Aedes albopictus and Culex pipiens mosquitoes in the context of native Wolbachia infections.

Microbial control of mosquitoes via the use of symbiotic or pathogenic microbes, such as Wolbachia and entomopathogenic fungi, are promising alternatives to synthetic insecticides to tackle the rapid increase in insecticide resistance and vector-borne disease outbreaks. This study evaluated the susceptibility and host responses of two important mosquito vectors, Ae. albopictus and Cx. pipiens, that naturally carry Wolbachia, to infections by entomopathogenic fungi. Our study indicated that while Wolbachia presence did not provide a protective advantage against entomopathogenic fungal infection, it nevertheless influenced the bacterial / fungal load and the expression of select anti-microbial effectors and phenoloxidase cascade genes in mosquitoes. Furthermore, although host responses from Ae. albopictus and Cx. pipiens were mostly similar, we observed contrasting phenotypes with regards to susceptibility and immune responses to fungal entomopathogenic infection in these two mosquitoes. This study provides new insights into the intricate multipartite interaction between the mosquito host, its native symbiont and pathogenic microbes that might be employed to control mosquito populations.

Enhancement the larvicidal activity of nanostructure copper oxide against Culex pipiens mosquito by yttrium replacement based on crystallite size reduction and topographic surface nature

The current work aims to improve the metal oxide characteristics for mosquito control. Un-doped and Y-doped CuO have been synthesized by simple chemical route. Structural, composition, and morphological properties were characterized by XRD, Raman spectra, EDX, SEM, and TEM techniques. The obtained results revealed that CuO was strongly affected by Y3+ support, in which the crystallite size decreased, and the surface area increased. Larvicidal performance was assessed against Culex pipiens suggesting that the nanocomposite CuO/Y of higher efficiency (LC50 = 7.67 mg /l, R2 = 0.977) compared with pure CuO. Light microscopy and SEM images exhibited larvae malformations owing to using the fabricated nanomaterials.

Mosquito abundance in relation to extremely high temperatures in urban and rural areas of Incheon Metropolitan City, South Korea from 2015 to 2020: an observational study

BackgroundDespite concerns regarding increasingly frequent and intense heat waves due to global warming, there is still a lack of information on the effects of extremely high temperatures on the adult abundance of mosquito species that are known to transmit vector-borne diseases. This study aimed to evaluate the effects of extremely high temperatures on the abundance of mosquitoes by analyzing time series data for temperature and mosquito abundance in Incheon Metropolitan City (IMC), Republic of Korea, for the period from 2015 to 2020.MethodsA generalized linear model with Poisson distribution and overdispersion was used to model the nonlinear association between temperature and mosquito count for the whole study area and for its constituent urban and rural regions. The association parameters were pooled using multivariate meta-regression. The temperature–mosquito abundance curve was estimated from the pooled estimates, and the ambient temperature at which mosquito populations reached maximum abundance (TMA) was estimated using a Monte Carlo simulation method. To quantify the effect of extremely high temperatures on mosquito abundance, we estimated the mosquito abundance ratio (AR) at the 99th temperature percentile (AR99th) against the TMA.ResultsCulex pipiens was the most common mosquito species (51.7%) in the urban region of the IMC, while mosquitoes of the genus Aedes (Ochlerotatus) were the most common in the rural region (47.8%). Mosquito abundance reached a maximum at 23.5 °C for Cx. pipiens and 26.4 °C for Aedes vexans. Exposure to extremely high temperatures reduced the abundance of Cx. pipiens mosquitoes {AR99th 0.34 [95% confidence interval (CI) 0.21–0.54]} to a greater extent than that of Anopheles spp. [AR99th 0.64 (95% CI 0.40–1.03)]. When stratified by region, Ae. vexans and Ochlerotatus koreicus mosquitoes showed higher TMA and a smaller reduction in abundance at extreme heat in urban Incheon than in Ganghwa, suggesting that urban mosquitoes can thrive at extremely high temperatures as they adapt to urban thermal environments.ConclusionsWe confirmed that the temperature-related abundance of the adult mosquitoes was species and location specific. Tailoring measures for mosquito prevention and control according to mosquito species and anticipated extreme temperature conditions would help to improve the effectiveness of mosquito-borne disease control programs.Graphical abstract

Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China.

Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.

Growth and Maintenance of Wolbachia in Insect Cell Lines

Simple SummaryWolbachia is an intracellular bacterium that occurs in arthropods and in filarial worms. First described nearly a century ago in the reproductive tissues of Culex pipiens mosquitoes, Wolbachia is now known to occur in roughly 50% of insect species, and has been considered the most abundant intracellular bacterium on earth. In insect hosts, Wolbachia modifies reproduction in ways that facilitate spread of the microbe within the host population, but otherwise is relatively benign. In this “gene drive” capacity, Wolbachia provides a tool for manipulating mosquito populations. In mosquitoes, Wolbachia causes cytoplasmic incompatibility, in which the fusion of egg and sperm nuclei is disrupted, and eggs fail to hatch, depending on the presence/absence of Wolbachia in the parent insects. Recent findings demonstrate that Wolbachia from infected insects can be transferred into mosquito species that do not host a natural infection. When transinfected into Aedes aegypti, an important vector of dengue and Zika viruses, Wolbachia causes cytoplasmic incompatibility and, in addition, decreases the mosquito’s ability to transmit viruses to humans. This review addresses the maintenance of Wolbachia in insect cell lines, which provide a tool for high-level production of infectious bacteria. In vitro technologies will improve use of Wolbachia for pest control, and provide the microbiological framework for genetic engineering of this promising biocontrol agent.The obligate intracellular microbe, Wolbachia pipientis (Rickettsiales; Anaplasmataceae), is a Gram-negative member of the alpha proteobacteria that infects arthropods and filarial worms. Although closely related to the genera Anaplasma and Ehrlichia, which include pathogens of humans, Wolbachia is uniquely associated with invertebrate hosts in the clade Ecdysozoa. Originally described in Culex pipiens mosquitoes, Wolbachia is currently represented by 17 supergroups and is believed to occur in half of all insect species. In mosquitoes, Wolbachia acts as a gene drive agent, with the potential to modify vector populations; in filarial worms, Wolbachia functions as a symbiont, and is a target for drug therapy. A small number of Wolbachia strains from supergroups A, B, and F have been maintained in insect cell lines, which are thought to provide a more permissive environment than the natural host. When transferred back to an insect host, Wolbachia produced in cultured cells are infectious and retain reproductive phenotypes. Here, I review applications of insect cell lines in Wolbachia research and describe conditions that facilitate Wolbachia infection and replication in naive host cells. Progress in manipulation of Wolbachia in vitro will enable genetic and biochemical advances that will facilitate eventual genetic engineering of this important biological control agent.

Target and non-target effects of Foeniculum vulgare and Matricaria chamomilla combined extract on Culex pipiens mosquitoes.

The present study focused on extracting green larvicides from extracts of the combination of Foeniculum vulgare and Matricaria chamomilla using different solvents of increasing polarity in a Soxhlet extractor and evaluating their ovicidal, larvicidal, and cytotoxic activities. The most promising among all tested extracts was hexane extract. The ovicidal activity of the hexane PH2 extract resulted in a significant (p < 0.05) decrease in egg hatchability from 95.00 ± 6.16% to 15 ± 9.04% at doses ranging from 62.5 to 500 µg/mL. The larval mortality with the hexane extract ranged from 13.33 ± 3.3% to 93.33 ± 3.3% at doses ranging from 31.25 to 250 µg/mL, respectively. The LC50 and LC90 values of the larvicidal activity of the hexane extract were estimated to be 148.3 and 242.17 µg/mL, respectively, after 24 h of exposure. Similarly, the LC50 values after 48 and 72 h of exposure were 124.93 and 100.3 µg/mL, respectively, against the third instar of Cx. pipiens. PH2 treatment of larvae resulted in histopathological changes such as degenerated epithelial cells and destruction of microvilli on the epithelial cells. The PH2 extract achieved a dose-dependent decrease in the rate of cell survival. The IC50 value of PH2-treated HUVECs was 192.07 µg/mL after 24 h of incubation. The cells showed changes in cellular and nuclear morphology. In conclusion, the hexane extract of PH2 could be used in mosquito management programs.

Short-Term Selection to Diflubenzuron and Bacillus thuringiensis Var. Israelensis Differentially Affects the Winter Survival of Culex pipiens f. Pipiens and Culex pipiens f. Molestus (Diptera: Culicidae).

Simple SummaryIn Europe, Culex pipiens (Diptera: Culicidae) mosquito, the prime vector of West Nile virus, consists of two forms, named pipiens and molestus, that exhibit substantial differences in their biology, including overwintering behavior. Diflubenzuron (DFB) and Bacillus thuringiensis var. israelensis (Bti) are among the most widely used larvicides which pose major concerns for resistance development. In temperate areas, winter represents a very challenging period for the survival of many insects, including mosquitoes, and therefore potential fitness costs associated with insecticide selection may reduce their overwintering success. In this context, we investigated how short-term selection of Cx. pipiens f. pipiens and molestus forms to DFB and Bti affect their overwintering success. Our findings revealed that selection to both larvicides induced a high fitness cost in terms of reduced winter survival of Cx. pipiens f. molestus but not of pipiens form, suggesting potential differences in the persistence of the selected individuals in the wild from year to year. The Culex pipiens (Diptera: Culicidae) mosquito is of high medical importance as it is considered the prime vector of West Nile virus. In Europe, this species consists of two forms, named pipiens and molestus, that exhibit substantial differences in their overwintering biology. Diflubenzuron (DFB) and Bacillus thuringiensis var. israelensis (Bti) are two of the most used larvicides in mosquito control, including that of Culex pipiens. The high dependency on these two larvicides poses major concerns for resistance development. The evolution and stability of resistance to insecticides has been associated with fitness costs that may be manifested under stressful conditions, such as the winter period. This study investigated how short-term selection of pipiens and molestus forms to both larvicides affect their overwintering success. Larvae from each form were subjected to the same selective pressure (80% mortality) for three successive generations with DFB and Bti. At the end of this process, the winter survival between the selected populations and the controls (colonies without selection) was determined for each form. Selection to both larvicides significantly reduced the winter survival rates of molestus but not of pipiens form, indicating potential differences in the persistence of the selected individuals from year to year between the two forms.

Assessment of West Nile Virus Lineage 2 Dynamics in Greece and Future Implications.

The emergence of West Nile Virus lineage 2 (WNV-2) has contributed to multiple major human outbreaks in Greece since 2010. Studies to date investigating biological and environmental factors that contribute to West Nile Virus (WNV) transmission have resulted in complex statistical models. We sought to examine open publicly available data to ascertain if a predictive risk assessment could be employed for WNV-2 in Greece. Based on accessible data, factors such as precipitation, temperature, and range of avian host species did not yield conclusive outcomes. However, by measuring the average rate of temperature change leading up to peak caseloads, we found a predictive characteristic to the timing of outbreaks. Detailed evolutionary studies revealed possible multiple introductions of WNV-2 in Europe, and that Greece acts through a source-sink inversion model, thereby allowing continued reseeding of WNV transmission each year by overwintering the Culex pipiens mosquito vector. Greece has proven an excellent model in WNV surveillance and has demonstrated the importance of rapid reporting for proper preparedness and response to vector-borne diseases.

Chemical Composition and Larvicidal Activity of Lavandula angustifolia Subsp. angustifolia and Lavandula dentata Spp. dentata Essential Oils against Culex pipiens Larvae, Vector of West Nile Virus

The Culex pipiens mosquito (Diptera: Culicidae) is highly suspected to be the vector responsible for the spread of several parasitic and viral diseases. The use of synthetic insecticides is generally the preferred method of controlling these mosquitoes’ proliferation. However, it has led to resistance problems in target mosquitoes and environmental damage. Hence, diverse plant extracts could be considered as an alternative and potential source as mosquito control agents. In this study, essential oils of Lavandula angustifolia subsp. angustifolia and Lavandula dentata spp. dentata that are growing in Morocco were examined for their insecticidal effects on Culex pipiens larvae. The bioassay was performed according to a methodology inspired by the standard protocol of the World Health Organization. The mortality rate was determined after 24 hours of exposure, and probit regression analysis was used to calculate LC50 and LC90. The chemical analysis revealed that the principal compounds of L. angustifolia subsp. essential oils include linalool, linalyl acetate, geraniol, lavandulyl acetate, camphor, β-caryophyllene, terpinen-4-ol, β-myrcene, and 1,8-cineole, while the essential oil of L. dentata spp. was mainly composed of 1,8-cineole, camphor, α-pinene, trans-pinocarveol, linalool, and borneol. These volatile compounds have shown a toxic effect against Culex pipiens larvae, with lethal concentrations LC50 and LC90 being, respectively, 140 µg/ml and 450 µg/ml, for the L. angustifolia subsp. essential oil. Meanwhile, they were estimated at 2670 µg/ml and 7400 µg/ml, respectively, for the L. dentata spp. essential oil. These results suggest using essential oils of two species of Lavandula to control the Culex pipiens mosquito. It could be useful for the study of new natural larvicidal compounds.