Evidence of natural Wolbachia infections and molecular identification of field populations of Culex pipiens complex (Diptera: Culicidae) mosquitoes in western Turkey

Establishing reliable risk projection information about the distribution pattern of members of the Culex pipiens complex is of particular interest, as these mosquitoes are competent vectors for certain disease‐causing pathogens. Wolbachia, a maternally inherited bacterial symbiont, are distributed in various arthropod species and can induce cytoplasmic incompatibility, i.e., reduced egg hatch, in certain crosses. It is being considered as a tool for population control of mosquito disease vectors. The Aegean region is characterized by highly populated, rural, and agricultural areas and is also on the route of the migratory birds. In this study, a fragment of the 658 bp of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene, which includes the barcode region, was employed to differentiate Cx. pipiens complex species found in this region. Moreover, for the first time, the prevalence of Wolbachia endobacteria in these natural populations was examined using PCR amplification of a specific wsp gene. Our results revealed a widespread (more than 90%, n=121) presence of the highly efficient West Nile virus vector Cx. quinquefasciatus in the region. We also found that Wolbachia infection is widespread; the average prevalence was 62% in populations throughout the region. This study provided valuable information about the composition of Cx. pipiens complex mosquitoes and the prevalence of Wolbachia infection in these populations in the Aegean region. This information will be helpful in tracking mosquito‐borne diseases and designing and implementing Wolbachia‐based control strategies in the region.     

Attraction of Culex pipiens to uropygial gland secretions does not explain feeding preference for American robins

Culex pipiens, the endemic mosquito vector of West Nile virus in eastern North America, is responsible for maintenance of the virus in avian reservoir hosts, the most important of which appears to be the American robin. One reason for the greater involvement of robins is believed to be the feeding preference of Cx. pipiens, however, the basis of this preference is not understood. We tested the hypothesis that the species‐specific chemical profile of avian uropygial gland secretions are used by Cx. pipiens as cues to locate birds and, therefore, may contribute to the observed feeding preferences. We used gas chromatography‐mass spectrometry to identify the semi‐volatile components of the uropygial gland secretions of American robins and two other common reservoir host species, the house sparrow and European starling. We found that the chemical composition of the robin secretions was different from those of the sparrows and starlings. Through behavioral choice trials conducted in a dual‐port olfactometer, we also found that Cx. pipiens did not prefer the secretions of robins over the other two species. Surprisingly, however, we found that Cx pipiens were more often attracted to live starlings over robins and to the secretions of starlings over those of robins.     

Mosquito traps for urban surveillance: collection efficacy and potential for use by citizen scientists

Mosquito‐borne diseases are a pervasive public health problem on a global scale, and effective management of them requires well‐designed surveillance programs for both vectors and pathogens. Mosquito traps are a common component of such programs, and their reach can be expanded by engaging citizen scientists. In this study in a southern Australian city, we compared the mosquito collection efficacy of two types of traps and assessed their suitability for use in citizen science programs. BG Sentinels and BG Gravid Aedes Trap (BG‐GAT) traps both collected Aedes and Culex species in similar proportions, albeit with the former collecting approximately nine times as many mosquitoes. However, BG Sentinels have a greater per unit cost than BG‐GATs and are restricted to deployment near power outlets. Importantly, despite being devised for collection of Aedes (Stegomyia) dengue vectors (such as Aedes aegypti), both traps can be effectively used in temperate climates for collection of a range of mosquito species. These traps could conceivably be used in citizen science programs to enhance the reach of surveillance at reduced cost.     

Screening of Brazilian Bacillus sphaericus strains for high toxicity against Culex quinquefasciatus and Aedes aegypti

Abstract:  In this work, 246 Bacillus sphaericus strains were evaluated against Aedes aegypti and Culex quinquefasciatus larvae to select the most effective ones to be used as the basis of a national product. All strains were isolated from different regions of Brazil and they are stored in a Bacillus spp. collection at Embrapa Genetic Resources and Biotechnology. The selected strains were characterized by biochemical and molecular methods. Based on selective bioassays, 87 strains were identified as toxic to one or both target species. All of these strains contain genes that encode the 42, 51 kDa proteins that constitute the binary toxin and the 100 kDa Mtx1 toxin. All toxic strains presented a very high LC₅₀ against A. aegypti , so, a product based on any of these B. sphaericus strains would not be recommended for use in programmes to control A. aegypti . S201 had highest activity against C. quinquefasciatus , presenting the lowest LC₅₀ and LC₉₀ in bioassays.     

香芹酮和薄荷醇手性异构体对淡色库蚊熏蒸及驱避活性

【目的】比较香芹酮和薄荷醇各手性异构体对淡色库蚊Culex pipiens pallens成蚊的熏蒸及驱避活性,为其应用提供理论依据。【方法】以3日龄淡色库蚊雌成蚊为供试昆虫,采用三角瓶熏蒸法比较测定香芹酮和薄荷醇的5种手性异构体的击倒与熏蒸致死活性;用Y-型嗅觉仪法比较测定驱避作用;将活性较好单体制成电热蚊香片,用方箱法进一步评价药效。【结果】5种异构体对淡色库蚊均具有一定的击倒和致死作用,其中L-香芹酮的击倒速度最快,KT50为9.75 min;D-香芹酮的致死作用最强,LC50为0.26μL/L。L-香芹酮和D-香芹酮对淡色库蚊均具较好的驱避作用,在0.5μL剂量下,处理30 min后的驱避率仍达100%。方箱法实验表明,以200μL的L-香芹酮和D-香芹酮为有效成分的电热蚊香片加热1 h后,对淡色库蚊雌成蚊的KT50分别为13.70和17.90 min。【结论】香芹酮对淡色库蚊雌成蚊的熏杀活性优于薄荷醇,且不同异构体的活性存在显著差异,L-香芹酮具备被开发为植物源卫生害虫防控剂的潜力。     

小蓬草精油对两种蚊虫的毒杀活性和成分分析

【目的】明确小蓬草Conyza canadensis ( L.) Cronq.精油的杀虫潜力及其活性成分。【方法】通过浸虫法和密闭熏蒸法测试了小蓬草精油对白纹伊蚊Aedes albopictus和致倦库蚊Culex pipiens quinquefasciatus幼虫及成蚊的毒杀活性, 并利用气相色谱 质谱联用仪（GC-MS）对精油的挥发性成分进行了定性分析。【结果】 小蓬草精油对白纹伊蚊1-4龄期幼虫及蛹的24 h LC50值分别为25.01, 45.88, 56.94, 64.60和346.23 μg/mL; 对致倦库蚊幼虫1-4龄期幼虫及蛹的24 h LC₅₀值分别为9.16, 8.65, 32.12, 43.68和197.83 μg/mL。在剂量分别为48, 64, 80, 96, 112和128 μg/cm3时, 小蓬草精油对白纹伊蚊成蚊的KT₅₀值分别为28.81, 22.31, 20.38, 17.05, 13.92和9.74 min; 对致倦库蚊的KT50值分别为34.90, 32.97, 23.97, 19.60, 15.20和10.34 min。 24 h熏蒸对白纹伊蚊和致倦库蚊成蚊的LC50值分别是75.46和99.19 μg/cm³。小蓬草精油的GC MS定性分析共分离鉴定出31种化合物, 其中单萜类物质6种, 倍半萜烯类物质17种, 含氧萜烯类6种。【结论】结果表明小蓬草精油对这两种蚊虫的毒杀活性较高, 具有深开发潜力。     

Evaluation of factors for rapid development of Culex quinquefasciatus in belowground stormwater treatment devices

Water samples from 11 belowground stormwater treatment Best Management Practices (BMPs) were evaluated for their capacity to support rapid development of the West Nile virus (WNV) mosquito vector, Culex quinquefasciatus. The observed minimum development time from egg to pupa ranged from six to over 30 days. Concentrations of potential food resources (total suspended solids and the particulate organic matter in water samples) were significantly correlated to development times. In addition, the rate of immature mosquito development was both site‐dependent and variable in time, suggesting that factors favorable to rapid development were strongly influenced by watershed characteristics and seasonal changes in temperature. Measured temperatures in belowground BMPs suggest that these structures may remain amenable to WNV virus activity longer each year than sites aboveground.     

Vector competence of Australian Culex gelidus Theobald (Diptera: Culicidae) for endemic and exotic arboviruses

The recent recognition of established populations of the mosquito, Culex gelidus Theobald, in Australia has raised concerns about local transmission of arboviruses. The vector competence of a mainland population of Cx. gelidus was investigated for two local alphaviruses, Ross River (RRV) and Barmah Forest (BFV) viruses, and three flaviviruses, Japanese encephalitis (JEV), Kunjin (KUNV) and Murray Valley encephalitis (MVEV) viruses. Colonised mosquitoes were exposed to virus via blood-soaked pledgets and transmission was tested using a capillary-tube method. The important Australian vectors, Aedes vigilax (Skuse) and Culex annulirostris Skuse, were used as internal controls for the alphaviruses and flaviviruses, respectively. Overall, Cx. gelidus was a more efficient vector of flaviviruses than alphaviruses. Culex gelidus was refractory to infection with BFV, and nearly 25% transmitted RRV, which was comparable to Ae. vigilax . Culex gelidus was susceptible to all three flaviviruses, with transmission rates of 96%, 95% and 41% for JEV, KUNV and MVEV, respectively. JEV transmission rates in Cx. annulirostris were unexpectedly low and this was possibly due to differences in susceptibility to JEV genotypes I and II. Considering the high susceptibility to the flaviviruses demonstrated here, and the natural infections with RRV and JEV that have been detected from northern Australian populations, the establishment of the exotic mosquito, Cx. gelidus , in Australia is potentially a significant public health concern.     

Identification of two mosquitocidal Bacillus cereus strains showing different host ranges

Mosquitocidal bacteria, M413 and C32 have been isolated from sediment samples collected from woodland and ditch, respectively. Gas chromatographic analysis of fatty acids methyl esters (GC-FAME) and 16S rRNA gene sequence alignment results showed these isolates belong to Bacillus cereus. The SDS-PAGE analysis of sporulated cultures of both isolates showed two major bands very similar in size. Interestingly, however, M413 is mainly toxic to 4th instars of Ochlerotatus taeniorhynchus whereas C32 is to those of Culex quinquefasciatus.