RESUMO
In arthropods, the intracellular bacteria Wolbachia often induce cytoplasmic incompatibility (CI) between sperm and egg, which causes conditional embryonic death and promotes the spatial spread of Wolbachia infections into host populations. The ability of Wolbachia to spread in natural populations through CI has attracted attention for using these bacteria in vector-borne disease control. The dynamics of incompatible Wolbachia infections have been deeply investigated theoretically, whereas in natural populations, there are only few examples described, especially among incompatible infected hosts. Here, we have surveyed the distribution of two molecular Wolbachia strains (wPip11 and wPip31) infecting the mosquito Culex pipiens in Tunisia. We delineated a clear spatial structure of both infections, with a sharp contact zone separating their distribution areas. Crossing experiments with isofemale lines from different localities showed three crossing types: wPip11-infected males always sterilize wPip31-infected females; however, while most wPip31-infected males were compatible with wPip11-infected females, a few completely sterilize them. The wPip11 strain was thus expected to spread, but temporal dynamics over 7 years of monitoring shows the stability of the contact zone. We examined which factors may contribute to the observed stability, both theoretically and empirically. Population cage experiments, field samples and modelling did not support significant impacts of local adaptation or assortative mating on the stability of wPip infection structure. By contrast, low dispersal probability and metapopulation dynamics in the host Cx. pipiens probably play major roles. This study highlights the need of understanding CI dynamics in natural populations to design effective and sustainable Wolbachia-based control strategies.
Assuntos
Culex/microbiologia , Genética Populacional , Wolbachia/genética , Animais , Técnicas de Tipagem Bacteriana , Cruzamentos Genéticos , Feminino , Masculino , Repetições de Microssatélites , Reprodução , Tunísia , Wolbachia/classificaçãoRESUMO
Retroelements represent a considerable fraction of many eukaryotic genomes and are considered major drives for adaptive genetic innovations. Recent discoveries showed that despite not normally using DNA intermediates like retroviruses do, Mononegaviruses (i.e., viruses with nonsegmented, negative-sense RNA genomes) can integrate gene fragments into the genomes of their hosts. This was shown for Bornaviridae and Filoviridae, the sequences of which have been found integrated into the germ line cells of many vertebrate hosts. Here, we show that Rhabdoviridae sequences, the major Mononegavirales family, have integrated only into the genomes of arthropod species. We identified 185 integrated rhabdoviral elements (IREs) coding for nucleoproteins, glycoproteins, or RNA-dependent RNA polymerases; they were mostly found in the genomes of the mosquito Aedes aegypti and the blacklegged tick Ixodes scapularis. Phylogenetic analyses showed that most IREs in A. aegypti derived from multiple independent integration events. Since RNA viruses are submitted to much higher substitution rates as compared with their hosts, IREs thus represent fossil traces of the diversity of extinct Rhabdoviruses. Furthermore, analyses of orthologous IREs in A. aegypti field mosquitoes sampled worldwide identified an integrated polymerase IRE fragment that appeared under purifying selection within several million years, which supports a functional role in the host's biology. These results show that A. aegypti was subjected to repeated Rhabdovirus infectious episodes during its evolution history, which led to the accumulation of many integrated sequences. They also suggest that like retroviruses, integrated rhabdoviral sequences may participate actively in the evolution of their hosts.
Assuntos
Artrópodes/genética , Evolução Molecular , Genoma de Inseto , Rhabdoviridae/genética , Animais , Teorema de Bayes , Bases de Dados Genéticas , Transferência Genética Horizontal , FilogeniaRESUMO
The α-proteobacteria Wolbachia are among the most common intracellular bacteria and have recently emerged as important drivers of arthropod biology. Wolbachia commonly act as reproductive parasites in arthropods by inducing cytoplasmic incompatibility (CI), a type of conditional sterility between hosts harboring incompatible infections. In this study, we examined the evolutionary histories of Wolbachia infections, known as wPip, in the common house mosquito Culex pipiens, which exhibits the greatest variation in CI crossing patterns observed in any insect. We first investigated a panel of 20 wPip strains for their genetic diversity through a multilocus scheme combining 13 Wolbachia genes. Because Wolbachia depend primarily on maternal transmission for spreading within arthropod populations, we also studied the variability in the coinherited Cx. pipiens mitochondria. In total, we identified 14 wPip haplotypes, which all share a monophyletic origin and clearly cluster into five distinct wPip groups. The diversity of Cx. pipiens mitochondria was extremely reduced, which is likely a consequence of cytoplasmic hitchhiking driven by a unique and recent Wolbachia invasion. Phylogenetic evidence indicates that wPip infections and mitochondrial DNA have codiverged through stable cotransmission within the cytoplasm and shows that a rapid diversification of wPip has occurred. The observed pattern demonstrates that a considerable degree of Wolbachia diversity can evolve within a single host species over short evolutionary periods. In addition, multiple signatures of recombination were found in most wPip genomic regions, leading us to conclude that the mosaic nature of wPip genomes may play a key role in their evolution.
Assuntos
Culex/microbiologia , Wolbachia/genética , Animais , Análise por Conglomerados , Culex/genética , DNA Mitocondrial/genética , Rearranjo Gênico , Marcadores Genéticos , Filogenia , Filogeografia , SimbioseRESUMO
Wolbachia are maternally inherited endosymbionts that can invade arthropod populations through manipulation of their reproduction. In mosquitoes, Wolbachia induce embryonic death, known as cytoplasmic incompatibility (CI), whenever infected males mate with females either uninfected or infected with an incompatible strain. Although genetic determinants of CI are unknown, a functional model involving the so-called mod and resc factors has been proposed. Natural populations of Culex pipiens mosquito display a complex CI relationship pattern associated with the highest Wolbachia (wPip) genetic polymorphism reported so far. We show here that C. pipiens populations from La Réunion, a geographically isolated island in the southwest of the Indian Ocean, are infected with genetically closely related wPip strains. Crossing experiments reveal that these Wolbachia are all mutually compatible. However, crosses with genetically more distant wPip strains indicate that Wolbachia strains from La Réunion belong to at least five distinct incompatibility groups (or crossing types). These incompatibility properties which are strictly independent from the nuclear background, formally establish that in C. pipiens, CI is controlled by several Wolbachia mod/resc factors.
Assuntos
Culex/microbiologia , Culex/fisiologia , Polimorfismo Genético , Simbiose , Wolbachia/genética , Wolbachia/fisiologia , Animais , Evolução Biológica , Cruzamentos Genéticos , Culex/genética , Citoplasma/fisiologia , DNA Mitocondrial/genética , Feminino , Marcadores Genéticos , Oceano Índico , Masculino , Mitose , Reação em Cadeia da Polimerase , Análise de Sequência de DNARESUMO
BACKGROUND: Insecticide resistance is a growing concern for malaria control and vector control effectiveness relies on assessing it distribution and understanding its evolution. METHODS: We assessed resistance levels and the frequencies of two major target-site mutations, L1014F-VGSC and G119S-ace-1, conferring resistance to pyrethroids (PYRs) and carbamates/organophosphates (CXs/OPs) insecticides. These data were compared to those acquired between 2006 and 2010 to follow resistance evolutionary trends over ten years. RESULTS: We report the results of a 3-year survey (2013-2015) of insecticide resistance in 13 localities across the whole country of Benin. Permethrin (PYR) resistance was found in all populations tested, L1014F-VGSC being almost fixed everywhere, while bendiocarb resistance was limited to a few localities, G119S-ace-1 remaining rare, with very limited variations during surveyed period. Interestingly, we found no effect of the type of insecticide pressure on the dynamics of these mutations. CONCLUSIONS: These results confirm both the high prevalence of PYR resistance and the potential of CXs/OPs as short- to medium-term alternatives in Benin. They also underline the need for regular resistance monitoring and informed management in their usage, as the G119S-ace-1 mutation is already present in Benin and surrounding countries. Their unwise usage would rapidly lead to its spread, which would jeopardize PYR-resistant Anopheles control.
Assuntos
Alelos , Anopheles/efeitos dos fármacos , Anopheles/genética , Resistência a Múltiplos Medicamentos/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Animais , Benin , Carbamatos/farmacologia , Evolução Molecular , Feminino , Proteínas de Insetos/genética , Malária , Controle de Mosquitos/métodos , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/genética , Mutação , Organofosfatos/farmacologia , Permetrina/farmacologia , Fenilcarbamatos , Piretrinas/farmacologiaRESUMO
BACKGROUND: Insecticide resistance is a rapid and recent evolutionary phenomenon with serious economic and public health implications. In the mosquito Anopheles gambiae s.s., main vector of malaria, resistance to organophosphates and carbamates is mainly due to a single amino-acid substitution in acetylcholinesterase 1 (AChE1). This mutation entails a large fitness cost. However, a resistant duplicated allele of the gene encoding AChE1 (ace-1), potentially associated to a lower fitness cost, recently appeared in An. gambiae. METHODS: Using molecular phenotype data collected from natural populations from West Africa, the frequency of this duplicated allele was investigated by statistical inference. This method is based on the departure from Hardy-Weinberg phenotypic frequency equilibrium caused by the presence of this new allele. RESULTS: The duplicated allele, Ag-ace-1(D), reaches a frequency up to 0.65 in Ivory Coast and Burkina Faso, and is potentially present in Benin. A previous study showed that Ag-ace-1(D), present in both M and S molecular forms in different West Africa countries, was generated by a single genetic event. This single origin and its present distribution suggest that this new allele is currently spreading. CONCLUSION: The spread of this less costly resistance allele could represent a major threat to public health, as it may impede An. gambiae control strategies, and thus increases the risk of malaria outbreaks.
Assuntos
Acetilcolinesterase/genética , Anopheles/enzimologia , Anopheles/genética , Resistência a Inseticidas/genética , Inseticidas , Controle de Mosquitos , Acetilcolinesterase/metabolismo , Alelos , Substituição de Aminoácidos , Animais , Anopheles/efeitos dos fármacos , Duplicação Gênica/efeitos dos fármacos , Frequência do Gene/efeitos dos fármacos , Genética Populacional , Genótipo , Insetos Vetores/efeitos dos fármacos , Insetos Vetores/enzimologia , Insetos Vetores/genética , Dados de Sequência Molecular , Mutação/efeitos dos fármacos , Mutação/genéticaRESUMO
In Tunisia, the mosquito Culex pipiens shows various organophosphate resistance alleles at Ester and ace-1 loci. The characterization and the distribution pattern of these alleles were studied among 20 populations sampled from north to center of Tunisia. At the Ester locus, Ester4, Ester5, and Ester(B12) were present. A new esterase characterized by the same electrophoretic migration as esterase A1 was identified: A13, encoded by Ester(A13) allele. At the ace-1 locus, the presence of the ace-1(R), ace-1(D), and F290V mutated alleles was also detected. A large heterogeneity in allelic frequencies at Ester and ace-1 loci was observed among samples, with a high significant genotypic differentiation considering both loci (F, = 0.077, P < 10(-5)), depicting variations of insecticide treatment intensity between areas. A comparison between populations collected in 1996 and 2005 showed an absence of significant resistance evolution. However, the high frequencies of resistance alleles in 2005 populations suggested that the selection pressures are still important in Tunisia. Strategies for resistance management are discussed in the context of the current knowledge of the Tunisian situation.
Assuntos
Culex/efeitos dos fármacos , Esterases/genética , Proteínas de Insetos/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Compostos Organofosforados/farmacologia , Animais , Culex/genética , Frequência do Gene , Geografia , TunísiaRESUMO
AIn the mosquito Culex pipiens (L.) (Diptera: Culicidae) esterases contribute to insecticide resistance by their increased activity. These esterases display a heterogeneous geographical distribution, particularly in Tunisia, where they are very diverse. In this study, we extended the characterization of a highly active esterase first detected in 1996: B12. Esterase B12 displayed the fastest electrophoretic mobility of all the previously described highly active esterases. We showed that it was encoded by the Ester(B12) allele at the Ester locus, and we isolated a strain, TunB12, homozygous for this allele. TunB12 displayed a low (approximately two- to three-fold) but significant resistance to the organophosphates temephos and chlorpyrifos, and to the pyrethroid permethrin. Only temephos resistance was synergized by S,S,S-tributyl-phosphorotrithioate. Real-time quantitative polymerase chain reaction revealed that the Ester(B12) allele was not amplified in TunB12 strain, indicating that B12 high activity could be due to a gene up-regulation mechanism. Ester(B12) allele frequencies also were estimated in 20 Tunisian populations collected in 2005. Analyses revealed a large distribution of this allele all over the country. Finally, sequences of Ester(B12) were acquired and genetic distance trees were constructed with the resistance Ester alleles already published, providing indications about allele's origins. The diverse array of highly active esterases in C. pipiens from Tunisia and the possible scenario of the origin of their coding alleles are discussed in the context of their possible evolution.
Assuntos
Culex/enzimologia , Esterases/metabolismo , Proteínas de Insetos/metabolismo , Animais , Culex/genética , Esterases/genética , Regulação da Expressão Gênica , Proteínas de Insetos/genética , Filogenia , TunísiaRESUMO
Acetylcholinesterase (AChE) is the target of two major insecticide families, organophosphates (OPs) and carbamates. AChE insensitivity is a frequent resistance mechanism in insects and responsible mutations in the ace gene were identified in two Diptera, Drosophila melanogaster and Musca domestica. However, for other insects, the ace gene cloned by homology with Drosophila does not code for the insensitive AChE in resistant individuals, indicating the existence of a second ace locus. We identified two AChE loci in the genome of Anopheles gambiae, one (ace-1) being a new locus and the other (ace-2) being homologous to the gene previously described in Drosophila. The gene ace-1 has no obvious homologue in the Drosophila genome and was found in 15 mosquito species investigated. In An. gambiae, ace-1 and ace-2 display 53% similarity at the amino acid level and an overall phylogeny indicates that they probably diverged before the differentiation of insects. Thus, both genes are likely to be present in the majority of insects and the absence of ace-1 in Drosophila is probably due to a secondary loss. In one mosquito (Culex pipiens), ace-1 was found to be tightly linked with insecticide resistance and probably encodes the AChE OP target. These results have important implications for the design of new insecticides, as the target AChE is thus encoded by distinct genes in different insect groups, even within the Diptera: ace-2 in at least the Drosophilidae and Muscidae and ace-1 in at least the Culicidae. Evolutionary scenarios leading to such a peculiar situation are discussed.
Assuntos
Acetilcolinesterase/genética , Acetilcolinesterase/metabolismo , Culicidae/enzimologia , Culicidae/genética , Proteínas de Drosophila/genética , Inseticidas/farmacologia , Sequência de Aminoácidos , Animais , Anopheles/enzimologia , Anopheles/genética , Sequência de Bases , Culex/enzimologia , Culex/genética , Culicidae/efeitos dos fármacos , Drosophila melanogaster/genética , Evolução Molecular , Genes de Insetos/genética , Resistência a Inseticidas/genética , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Two groups of Anopheles claviger sensu stricto Meigen (Diptera: Culicidae) were recently found in France, representing unclear genetic entities. To better understand this situation, sampling was extended to 13 European countries, and 47 samples were analyzed by investigating the polymorphism of 11 autosomal and 1 sex-linked allozyme loci. Genetic differentiation, as measured by F(st), between these two groups was confirmed, with no isolation by distance within each group. Among the twelve loci studied, none had diagnostic alleles. Group I is mainly located in western Europe (UK and south-west of France), and Group II covers eastern France and eastern and northern Europe. Intermediate populations, sampled at the overlapping range between them, do not display a heterozygote deficit, suggesting that these two groups are probably not genetically isolated. The origin of each group and its biological significance is discussed within the context of differentiation in refugia during recent glaciations.
Assuntos
Anopheles/classificação , Anopheles/genética , Polimorfismo Genético , Animais , Anopheles/crescimento & desenvolvimento , Europa (Continente) , Geografia , Isoenzimas/genética , Larva , Filogenia , PupaRESUMO
This study was aimed at clarifying the nature of the resistance to Bacillus sphaericus Neide (Bs) that Culex pipiens L. has developed in west Mediterranean countries, France, and Tunisia. Two recessive and sex-linked mutants, sp-1R and sp-2R, were previously detected in southern France. Here, the Tunisian resistance was also shown to involve a single recessive and sex-linked gene that was temporarily named sp-T(R). In addition, sp-1R, sp-2R, and sp-T(R) were shown to separately confer a similar high resistance level (> 5,000-fold) in the homozygous state. Knowing that sp-1R resistance does not alter the binding of Bs binary toxin to its specific receptor, we investigated this character in sp-2RR and sp-T(RR) homozygotes. This was performed by in vitro experiments in which larval brush border membrane fractions (BBMF) were exposed to the 125I-Bin2 toxin of B. sphaericus strain 1593. The toxin-receptor binding was found disrupted by sp-2R but not by sp-T(R). Comparing the binding kinetics among nine Culex pipiens strains of diverse origins revealed that the Bs receptors of sp-1RR and Sp-T(RR) homozygous larvae were displaying the highest affinity toward Bs binary toxins. These results are discussed with regard to alternative assumptions on the dynamics of high Bs-resistance and on the emerging possibilities to test them in a near future.
Assuntos
Bacillus , Toxinas Bacterianas/farmacologia , Culex/efeitos dos fármacos , Inseticidas/farmacologia , Animais , Toxinas Bacterianas/metabolismo , Culex/genética , Feminino , França , Resistência a Inseticidas/genética , Inseticidas/metabolismo , Masculino , Região do Mediterrâneo , Receptores de Superfície Celular/metabolismo , TunísiaRESUMO
Population surveys of Culex pipiens quinquefasciatus carried out in 1991 and 1999 were compared with data collected in 1990 before the beginning of the control program against this mosquito. Larval samples collected in 1999 displayed resistance to the four tested insecticides: permethrin, propoxur, temephos, and chlorpyrifos. Temephos resistance ratio at LC50 (RR50) ranged between 8.1- and 42-fold compared with 2.9- and 4.6-fold in 1990, and chlorpyrifos RR50 ranged between 8.6- and 123-fold compared with 6.4- and 19-fold in 1990. This increased resistance to organophosphorus insecticides was associated with a sharp decrease of susceptible genotypes at two loci (Ester and ace-1), as well as to an allele replacement at the Ester locus.
Assuntos
Culex , Resistência a Medicamentos , Inseticidas/toxicidade , Animais , Clorpirifos/toxicidade , Culex/classificação , Culex/genética , Genótipo , Geografia , Martinica , Permetrina/toxicidade , Fenótipo , Densidade Demográfica , Propoxur/toxicidade , Temefós/toxicidadeRESUMO
Gene duplications have long been advocated to contribute to the evolution of new functions. The role of selection in their early spread is more controversial. Unless duplications are favored for a direct benefit of increased expression, they are likely detrimental. In this article, we investigated the case of duplications favored because they combine already functionally divergent alleles. Their gene-dosage/fitness relations are poorly known because selection may operate on both overall expression and duplicates relative dosage. Using the well-documented case of Culex pipiens resistance to insecticides, we compared strains with various ace-1 allele combinations, including two duplicated alleles carrying both susceptible and resistant copies. The overall protein activity was nearly additive, but, surprisingly, fitness correlated better with the relative proportion of susceptible and resistant copies rather than any absolute measure of activity. Gene dosage is thus crucial, duplications stabilizing a "heterozygote" phenotype. It corroborates the view that these were favored because they fix a permanent heterosis, thereby solving the irreducible trade-off between resistance and synaptic transmission. Moreover, we showed that the contrasted successes of the two duplicated alleles in natural populations depend on genetic changes unrelated to ace-1, confirming the probable implication of recessive sublethal mutations linked to structural rearrangements in some duplications.
Assuntos
Culex/genética , Dosagem de Genes , Duplicação Gênica , Aptidão Genética , Resistência a Inseticidas/genética , Animais , Culex/efeitos dos fármacos , Culex/metabolismo , Evolução Molecular , Genes de Insetos , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Inseticidas/farmacologia , Mutação , Fenótipo , Temefós/farmacologiaRESUMO
The status of genes conferring resistance to organophosphate and carbamate insecticides has been examined in Culex pipiens pipiens mosquitoes sampled in Algeria. Presence of overproduced esterases was sporadic, but acetylcholinesterase-1 resistant alleles were observed in almost all samples. We focused our study on the AChE1 G119S substitution characterized in almost all samples, mostly at the heterozygous state. A genetic test revealed the presence of ace-1 duplication associating a susceptible and a resistant ace-1 copy. Molecular characterization showed a high occurrence of ace-1 duplication with six distinct duplicated alleles out of four samples. The inferred frequency of duplicated allele suggests that it is replacing the single resistant G119S allele. Finally, we discuss the mechanism at the origin of these duplicated haplotypes and their consequences on the management of insecticide resistance.
Assuntos
Acetilcolinesterase/genética , Culex/genética , Duplicação Gênica , Proteínas de Insetos/genética , Resistência a Inseticidas/genética , Acetilcolinesterase/metabolismo , Argélia , Alelos , Substituição de Aminoácidos , Animais , Culex/efeitos dos fármacos , Culex/metabolismo , Feminino , Frequência do Gene , Haplótipos , Heterozigoto , Incidência , Proteínas de Insetos/metabolismo , Resistência a Inseticidas/efeitos dos fármacos , Inseticidas/farmacologia , Masculino , Organofosfatos/farmacologia , FilogeniaRESUMO
The use of the bacterium Wolbachia is an attractive alternative method to control vector populations. In mosquitoes, as in members of the Culex pipiens complex, Wolbachia induces a form of embryonic lethality called cytoplasmic incompatibility, a sperm-egg incompatibility occurring when infected males mate either with uninfected females or with females infected with incompatible Wolbachia strain(s). Here we explore the feasibility of the Incompatible Insect Technique (IIT), a species-specific control approach in which field females are sterilized by inundative releases of incompatible males. We show that the Wolbachia wPip(Is) strain, naturally infecting Cx. p. pipiens mosquitoes from Turkey, is a good candidate to control Cx. p. quinquefasciatus populations on four islands of the south-western Indian Ocean (La Réunion, Mauritius, Grande Glorieuse and Mayotte). The wPip(Is) strain was introduced into the nuclear background of Cx. p. quinquefasciatus mosquitoes from La Réunion, leading to the LR[wPip(Is)] line. Total embryonic lethality was observed in crosses between LR[wPip(Is)] males and all tested field females from the four islands. Interestingly, most crosses involving LR[wPip(Is)] females and field males were also incompatible, which is expected to reduce the impact of any accidental release of LR[wPip(Is)] females. Cage experiments demonstrate that LR[wPip(Is)] males are equally competitive with La Réunion males resulting in demographic crash when LR[wPip(Is)] males were introduced into La Réunion laboratory cages. These results, together with the geographic isolation of the four south-western Indian Ocean islands and their limited land area, support the feasibility of an IIT program using LR[wPip(Is)] males and stimulate the implementation of field tests for a Cx. p. quinquefasciatus control strategy on these islands.
Assuntos
Culex/crescimento & desenvolvimento , Culex/microbiologia , Controle de Mosquitos/métodos , Controle Biológico de Vetores/métodos , Wolbachia/crescimento & desenvolvimento , Animais , Feminino , Ilhas do Oceano Índico , Infertilidade , Masculino , Análise de SobrevidaRESUMO
Members of the Anopheles gambiae complex are among the best malaria vectors in the world, but their vectorial capacities vary between species and populations. A large-scale sampling of An. gambiae sensu lato was carried out in 2006 and 2007 in various bioclimatic areas of Benin (West Africa). The objective of this study was to collate data on the relative frequencies of species and forms within the An. gambiae complex and to produce a map of their spatial distribution. Sampling took place at 30 sites and 2122 females were analyzed. Two species were identified through molecular methods. The overall collection showed a preponderance of An. gambiae s.s., but unexpectedly, An. arabiensis was reported in the coastal-Guinean bioclimatic area characterized by a mean annual rainfall of >1500 mm where only An. gambiae s.s. was reported previously. Our study of Benin indicates that An. arabiensis would be adapted not only to the urban areas but also to the rural humid regions. Among 1717 An. gambiae s.s., 26.5% were of the M form and 73.3% were S form. Few hybrid specimens between the M and S forms were observed (0.2%). Only the spatial distribution of the M form appears to be mainly a function of bioclimatic area. Factors that influence the distribution of these malaria vectors are discussed. This study underlines the need of further investigations of biological, ecological, and behavioral traits of these species and forms to better appreciate their vectorial capacities. Acquisition of entomological field data appears essential to better estimate the stratification of malaria risk and help improve malaria vector control interventions.
Assuntos
Anopheles/classificação , Anopheles/genética , Vetores de Doenças , Animais , Anopheles/crescimento & desenvolvimento , Benin , Quimera , GeografiaRESUMO
Resistance to insecticides was monitored on Culex pipiens quinquefasciatus mosquitoes collected in twelve localities of La Réunion, a geographically isolated island of the Indian Ocean. This mosquito is of medical concern in the region as a known vector for filariasis and a potential vector for West Nile and Rift Valley Fever viruses. Our bioassays indicated the presence of resistance to all tested insecticides, i.e. organochlorides, organophosphates and pyrethroids. A molecular investigation revealed a higher frequency of resistance genes in the coastal areas compared to elevated rural sites, probably reflecting the different nature of insecticide pressures together with the genetic cost of resistance alleles. A simple molecular test was developed to detect Rdl allele, encoding a gamma-aminobutyric acid (GABA) receptor resistant to dieldrin. Unexpectedly high Rdl frequencies were recorded over the whole island, despite this insecticide having been banned for over 15 years. This resistant allele was also detected for the first time in two samples of Aedes albopictus, a species recently involved in severe Chikungunya epidemics on the island. Rdl selection in these two mosquito species discloses current insecticide pressures in urban areas, from unknown origins, that should be taken into account to develop vector control strategies.
Assuntos
Aedes/genética , Culex/genética , Resistência a Inseticidas/genética , Acetilcolinesterase/genética , Alelos , Animais , Sequência de Bases , Dieldrin , Feminino , Frequência do Gene , Genes de Insetos , Inseticidas , Masculino , Dados de Sequência Molecular , Organofosfatos , Piretrinas , Receptores de GABA/genética , Reunião , Alinhamento de SequênciaRESUMO
Two amino acid substitutions in acetylcholinesterase 1 (AChE1), G119S and F290V, are responsible for resistance to organophosphate and carbamate insecticides in Culex pipiens mosquitoes. These mutations generate very different levels of insensitivity to insecticide inhibitors. We described here a biochemical method that rapidly identifies AChE1 variants (susceptible, G119S and F290V, named S, R and V, respectively) present in individual mosquitoes. We investigated the frequency of AChE1 phenotypes in 41 field samples collected around the Mediterranean Sea. F290V substitution was found only in 15 samples and at low frequency, whereas G119S was highly spread in all samples. However, seven V distinct alleles were identified whereas only one R allele was present. The [V] enzymatic phenotype was never observed alone, and the V allele was always found associated with the susceptible and/or G119S AChE1 ([VS], [VR] or [VRS] phenotypes). Furthermore, we showed the presence of duplicated alleles, associating a susceptible and a V copy of the ace-1 gene, in most individuals analyzed for its presence. Evolutionary forces driving the large number of F290V ace-1 alleles and their low frequency in Mediterranean countries are discussed.
Assuntos
Acetilcolinesterase/genética , Acetilcolinesterase/metabolismo , Culex/efeitos dos fármacos , Culex/enzimologia , Acetilcolinesterase/química , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Sequência de Bases , Culex/fisiologia , Demografia , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Filogenia , Fatores de TempoRESUMO
Alkaline phosphatases and several dehydrogenases and oxidases separated by a microdisc electrophoresis technique have been studied during larval and early pupal development ofD. pseudoobscura salivary glands, fat body, hemolymph, body wall and whole body. Tissue-specific enzymes were observed and the qualitative differences occurring during the development are discussed.