Search

Abstract: In natural populations of mosquitoes, high level of resistance to carbamates (CX) and organophosphates (OP) is provided by insensitive acetylcholinesterase (AChE1). Different alleles conferring resistance have been identified at the ace1 locus. They differ from the wild-type by only one amino-acid substitution. The comparison of the biochemical characteristics of mutated recombinant proteins and AChE1 in resistant mosquito extracts confirmed the role of each substitution in insensitivity. Selection of these different resistant alleles in field populations depends likely on the insecticides used locally. Theoretical modelling studies are initiated to develop novel strategies of mosquito control. [Copyright &y& Elsevier]

Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and have evolved several different ways for manipulating their host, the most frequent being cytoplasmic incompatibility (CI). CI leads to embryo death in crosses between infected males and uninfected females, as well as in crosses between individuals infected by incompatible Wolbachia strains. In the mosquito Culex pipiens, previous studies suggested developmental variation in embryos stemming from different incompatible crosses. We have investigated this variation in different incompatible crosses. Unhatched eggs were separated into three classes based upon the developmental stage reached by the embryos. We found that incompatible crosses involving uninfected females produced only embryos whose development was arrested at a very early stage, irrespective of the Wolbachia variant infecting the male. These results differ from other host species where a developmental gradient that could reach late stages of embryogenesis or even living larvae was observed, and indicate a novel peculiarity of CI mechanism in C. pipiens. By contrast, all incompatible crosses with infected C. pipiens females produced embryos of all three classes. The proportion of embryo classes appeared to be associated with the strains involved, suggesting specific CI properties in different incompatible crosses. In addition, the contribution of parental genome was characterized in embryo classes using molecular markers for each chromosome. Embryo phenotypes appeared linked to the paternal chromosomes' contribution, as described in Drosophila simulans. However, this contribution varied according to maternal infection and independently of male factors.Heredity (2006) 96, 493–500. doi:10.1038/sj.hdy.6800831; published online 26 April 2006 [ABSTRACT FROM AUTHOR]

High insecticide resistance resulting from insensitive acetylcholinesterase (AChE) has emerged in mosquitoes. A single mutation (G119S of the ace-1 gene) explains this high resistance in Culex pipiens and in Anopheles gambiae. In order to provide better documentation of the ace-1 gene and the effect of the G119S mutation, we present a three-dimension structure model of AChE, showing that this unique substitution is localized in the oxyanion hole, explaining the insecticide insensitivity and its interference with the enzyme catalytic functions. As the G119S creates a restriction site, a simple PCR test was devised to detect its presence in both A. gambiae and C. pipiens, two mosquito species belonging to different subfamilies (Culicinae and Anophelinae). It is possibile that this mutation also explains the high resistance found in other mosquitoes, and the present results indicate that the PCR test detects the G119S mutation in the malaria vector A. albimanus. The G119S has thus occurred independently at least four times in mosquitoes and this PCR test is probably of broad applicability within the Culicidae family. [ABSTRACT FROM AUTHOR]

Anopheles gambiaes.s. is a complex of sibling taxa characterized by various paracentric inversions. In west and central Africa, where several taxa are sympatric, a kdr mutation responsible for pyrethroid resistance has been described in only one (the S taxon), suggesting an absence of gene flow between them. Following a thorough sampling, we have found a kdr mutation in another taxon (M). To establish whether this mutation is the same event or not, the large intron upstream of the kdr mutation was sequenced to find polymorphic sites in susceptible/resistant and M/S mosquitoes. The low genetic diversity found in this DNA region indicates that a local genetic sweep has recently occurred. However, some polymorphic sites were found, and it is therefore concluded that the kdr mutation in the M taxon is not an independent mutation event, and is best explained by an introgression from the S taxon. These results are discussed within the context of possible gene flow between members of An. gambiae s.s. taxa, and with the possible spread of the kdr mutation in other closely related malaria vectors of the An. gambiae complex. [ABSTRACT FROM AUTHOR]

Presents a corrected reprint of an article previously published in the periodical 'Nature.'

Maternally inherited Wolbachia often manipulate the reproduction of arthropods to promote their transmission. In most species, Wolbachia exert a form of conditional sterility termed cytoplasmic incompatibility (CI), characterized by the death of embryos produced by the mating between individuals with incompatible Wolbachia infections. From a theoretical perspective, no stable coexistence of incompatible Wolbachia infections is expected within host populations and CI should induce the invasion of one strain or of a set of compatible strains. In this study, we investigated this prediction on CI dynamics in natural populations of the common house mosquito Culex pipiens. We surveyed the Wolbachia diversity and the expression of CI in breeding sites of the south of France between 1990 and 2005. We found that geographically close C. pipiens populations harbor considerable Wolbachia diversity, which is stably maintained over 15 years. We also observed a very low frequency of infertile clutches within each sampled site. Meanwhile, mating choice experiments conducted in laboratory conditions showed that assortative mating does not occur. Overall, this suggests that a large set of compatible Wolbachia strains are always locally dominant within mosquito populations thus, fitting with the theoretical expectations on CI dynamics. [ABSTRACT FROM AUTHOR]

Wolbachia are maternally inherited endocellular bacteria, widespread in invertebrates and capable of altering several aspects of host reproduction. Cytoplasmic incompatibility (CI) is commonly found in arthropods and induces hatching failure of eggs from crosses between Wolbachia-infected males and uninfected females (or females infected by incompatible strains). Several factors such as bacterial and host genotypes or bacterial density contribute to CI strength and it has been proposed, mostly from Drosophila data, that older males have a lower Wolbachia load in testes which, thus, induces a lighter CI. Here, we challenge this hypothesis using different incompatible Culex pipiens mosquito strains and show that CI persists at the same intensity throughout the mosquito life span. Embryos from incompatible crosses showed even distributions of abortive phenotypes over time, suggesting that host ageing does not reduce the sperm-modification induced by Wolbachia. CI remained constant when sperm was placed in the spermathecae of incompatible females, indicating that sperm modification is also stable over time. The capacity of infected females to rescue CI was independent of age. Last, the density of Wolbachia in whole testes was highly strain-dependent and increased dramatically with age. Taken together, these data stress the peculiarity of the C.pipiens/Wolbachia interaction and suggest that the bacterial dosage model should be rejected in the case of this association.Heredity (2007) 98, 368–374. doi:10.1038/sj.hdy.6800948; published online 7 March 2007 [ABSTRACT FROM AUTHOR]

To obtain a good estimation of particle size distribution and concentration encountered by car passengers during their journeys, an electrical low-pressure impactor was installed in a car. Two measurement campaigns were carried out. The first one covered various traffic conditions in urban and rural areas near Rouen (France). The second campaign covered several journeys up and down the same tunnel. Measurements were recorded in the range 30 nm–10 μm at 1 Hz frequency.The size distributions show two main shapes depending mainly on the concentration. For high concentrations the distributions correspond to diesel emissions with the mode comprised between 60 and 100 nm. For low concentrations the maximum of the distribution is located in the finest particle class of the device, between 30 and 60 nm.The concentrations observed are generally less than 50 000 part cm−3 but may reach more than 106 part cm−3 during specific road events. Likewise high-concentration values may be regularly observed at the end of the tunnel along which the concentration increases linearly, after a surprising decrease. One main conclusion is that on-road measurements are necessary for a correct evaluation of the conditions encountered by car passengers. [Copyright &y& Elsevier]

Graphical abstract Frequency of the diflubenzuron (DFB) susceptible and resistant alleles found in the Emilia-Romagna provinces (Italy) associated with DFB pressures from mosquito and orchard pest controls. Highlights • Diflubenzuron resistant mutations occur in Italian populations of Culex pipiens , reaching over 70%. • Resistant allele frequencies increased from West to East of Emilia-Romagna region. • The intense insecticide pressure due to past agricultural applications, along with current mosquito control, can account for the observed pattern of focal resistance. • Ongoing Cx. pipiens resistance surveillance is of utmost importance to prevent its diffusion. Abstract Insecticide resistance is a major threat for vector control and prevention of mosquito borne diseases. In the Culex pipiens mosquitoes, resistance against diflubenzuron (DFB) was firstly detected in Ravenna (Emilia-Romagna region, Northern Italy), in 2015. The resistant phenotypes were associated with two mutations, I1043 M and I1043 L, at the amino acid 1043 of the chitin synthase gene. In this study, we monitored the presence, frequency and geographical distribution of the DFB resistant mutations in Cx. pipiens populations from Northern Italy, and in populations from Greece and France. In the Emilia-Romagna region, the resistant mutations were detected in 20 out of the 30 populations analysed, reaching allelic frequencies over 70%. The presence and distribution of the resistance mutations was highly focal, with a clear pattern of increasing resistant allelic frequencies moving from the Western towards the Eastern provinces of Emilia-Romagna. Contrary to Italy, DFB resistant alleles were not detected in the Cx. pipiens mosquitoes sampled from Greece and France. Following statistical, literature and bibliographical database analyses on the history of DFB insecticide use in the study areas, we suggest that the selection pressures from the intense agricultural DFB applications occurring throughout the' 80-'90 s against orchard pests, followed, from 2000s onwards by mosquito control DFB applications, may account for the high mutation frequencies observed in the Cx. pipiens populations of the Eastern provinces of Emilia-Romagna. The findings are of major concern for public health in Italy and Europe, as DFB remains a very important insecticide used for controlling arbovirus mosquito vectors, where alternative larvicides are extremely limited. [ABSTRACT FROM AUTHOR]

The Culex pipiens mosquito complex is a group of evolutionarily closely related species including C. pipiens and Culex quinquefasciatus, both infected by the cytoplasmically inherited Wolbachia symbiont. A Wolbachia-uninfected population of C. pipiens was however described in South Africa and was recently proposed to represent a cryptic species. In this study, we reconsidered the existence of this species by undertaking an extensive screening for the presence of Wolbachia-uninfected C. pipiens specimens and by characterizing their genetic relatedness with known members of the complex. We first report on the presence of Wolbachia-uninfected specimens in several breeding sites. We next confirm that these uninfected specimens unambiguously belong to the C. pipiens complex. Remarkably, all uninfected specimens harbour mitochondrial haplotypes that are either novel or identical to those previously found in South Africa. In all cases, these mitochondrial haplotypes are closely related, but different, to those found in other C. pipiens complex members known to be infected by Wolbachia. Altogether, these results corroborate the presence of a widespread cryptic species within the C. pipiens species complex. The potential role of this cryptic C. pipiens species in the transmission of pathogens remains however to be determined. The designation ' Culex juppi nov. sp.' is proposed for this mosquito species. [ABSTRACT FROM AUTHOR]

Current views about the impact of Wolbachia on Plasmodium infections are almost entirely based on data regarding artificially transfected mosquitoes. This work has shown that Wolbachia reduces the intensity of Plasmodium infections inmosquitoes, raising the exciting possibility of using Wolbachia to control or limit the spread of malaria. Whether natural Wolbachia infections have the same parasite-inhibiting properties is not yet clear. Wolbachia�mosquito combinations with a long evolutionary history are, however, key for understanding what may happen with Wolbachia-transfected mosquitoes after several generations of coevolution. We investigate this issue using an entirely natural mosquito�Wolbachia�Plasmodium combination. In contrast to most previous studies, which have been centred on the quantification of the midgut stages of Plasmodium, we obtain a measurement of parasitaemia that relates directly to transmission by following infections to the salivary gland stages. We show that Wolbachia increases the susceptibility of Culex pipiens mosquitoes to Plasmodium relictum, significantly increasing the prevalence of salivary gland stage infections. This effect is independent of the density of Wolbachia in the mosquito. These results suggest that naturally Wolbachia-infected mosquitoes may, in fact, be better vectors of malaria than Wolbachia-free ones. [ABSTRACT FROM AUTHOR]

Benin has embraced World Health Organization-recommended preventive strategies to control malaria. Its National Malaria Control Programme is implementing and/or coordinating various actions and conducting evaluation trials of mosquito control strategies. Mosquito control is based on the use of insecticide-treated nets and indoor residual spraying, but the efficacy of these strategies to control malaria vectors is endangered by insecticide resistance. Here, we present the results of a nationwide survey on the status of insecticide susceptibility and resistance in Anopheles gambiae s.l. (Diptera: Culicidae) carried out in Benin in 2006-2007 (i.e. before extensive vector control was undertaken). Overall, our study showed that the S molecular form of An. gambiae s.s. predominates and is widely distributed across the country, whereas the frequency of the M form shows a strong decline with increasing latitude. Susceptibility to DDT, permethrin, carbosulfan and chlorpyrifos-methyl was assessed; individual mosquitoes were identified for species and molecular forms, and genotyped for the kdr and ace-1 loci. Full susceptibility to chlorpyrifos-methyl was recorded and very few samples displayed resistance to carbosulfan. High resistance levels to permethrin were detected in most samples and almost all samples displayed resistance to DDT. The kdr-Leu-Phe mutation was present in all localities and in both molecular forms of An. gambiae s.s. Furthermore, the ace-1 R mutation was predominant in the S form, but absent from the M form. By contrast, no target modification was observed in Anopheles arabiensis. Resistance in the An. gambiae S molecular form in this study seemed to be associated with agricultural practices. Our study showed important geographic variations which must be taken into account in the vector control strategies that will be applied in different regions of Benin. It also emphasizes the need to regularly monitor insecticide resistance across the country and to adapt measures to manage resistance. [ABSTRACT FROM AUTHOR]

The maternally inherited symbiotic Wolbachia have been previously shown to have much greater densities in insecticide-resistant Culex pipiens mosquitoes than in insecticide-susceptible individuals. These high densities were shown to be at least partially responsible for the costs related to insecticide resistance in this species. We report here the rapid evolution, on the order of 50 generations, of bacterial densities both in laboratory and field populations. Along with other recently published studies, this report shows that Wolbachia–host interactions are very dynamic. [ABSTRACT FROM AUTHOR]

A severe Chikungunya (CHIK) outbreak recently hit several countries of the Indian Ocean. On La Réunion Island, Aedes albopictus was incriminated as the major vector. This mosquito species is naturally co-infected with two distinct strains of the endosymbiont Wolbachia, namely wAlbA and wAlbB, which are increasingly attracting interest as potential tools for vector control. A PCR quantitative assay was developed to investigate Wolbachia/mosquito host interactions. We show that Wolbachia densities are slightly decreased in CHIK virus (CHIKV)-infected females. We measured the impact of CHIKV replication on a lysogenic virus: WO bacteriophage. Our data indicate that WO is sheltered by wAlbB, likely at a single copy per bacteria, and that CHIKV replication is not a physiological stress triggering WO entrance into the lytic cycle. [ABSTRACT FROM AUTHOR]

In most parts of China, mosquitoes have been subjected to organophosphate (OP) insecticide treatments since the mid-1960s, and resistance gene monitoring in the Culex pipiens complex (Diptera: Culicidae) started in only a few locations from the end of the 1980s. Many resistant alleles at the Ester locus have been found in field populations, including those commonly found around the world ( Ester B1 and Ester 2), and those endemic to China ( Ester B6, Ester B7, Ester 8, and Ester 9). This situation is atypical, and may represent a complex situation for the evolution of insecticide resistance genes in China. To increase our understanding of the Chinese situation and our ability to manage resistance in the C. pipiens complex, a large study was performed. Twenty field populations were sampled from Beijing to Guangzhou. Bioassays with five insecticides (dichlorvos, parathion, chlorpyrifos, 2-sec-butylphenyl methyl carbamate, and propoxur) disclosed resistance levels variable according to the geographic origin, and up to 85-fold for dichlorvos. Six overproduced esterases were identified, including two that have not been previously described. Most of them were found in all samples, although at variable frequencies, suggesting variable selection or a transient situation, e.g., each one was recently restricted to a particular geographic area. The results are discussed in the context of recent alterations to insecticide campaigns, and of the evolution of resistance genes in Chinese C. pipiens populations. [ABSTRACT FROM AUTHOR]

Microsatellite markers and chromosomal inversion polymorphisms are useful genetic markers for determining population structure in Anopheline mosquitoes. In Anopheles funestus (2N = 6), only chromosome arms 2R, 3R, and 3L are known to carry polymorphic inversions. The physical location of microsatellite markers with respect to polymorphic inversions is potentially important information for interpreting population genetic structure, yet none of the available marker sets have been physically mapped in this species. Accordingly, we mapped 32 polymorphic A. funestus microsatellite markers to the polytene chromosomes using fluorescent in situ hybridization (FISH) and identified 16 markers outside of known polymorphic inversions. Here we provide an integrated polytene chromosome map for A. funestus that includes the breakpoints of all known polymorphic inversions as well as the physical locations of microsatellite loci developed to date. Based on this map, we suggest a standard set of 16 polymorphic microsatellite markers that are distributed evenly across the chromosome complement, occur predominantly outside of inversions, and amplify reliably. Adoption of this set by researchers working in different regions of Africa will facilitate metapopulation analyses of this primary malaria vector. [ABSTRACT FROM AUTHOR]