Dataset Information

Susceptibility Status and Resistance Mechanisms in Permethrin-Selected, Laboratory Susceptible and Field-Collected Aedes aegypti from Malaysia.

ABSTRACT: This study is intended to provide a comprehensive characterization of the resistance mechanisms in the permethrin-selected (IMR-PSS) and laboratory susceptible (IMR-LS) Aedes aegypti strain from Malaysia. Both IMR-PSS and IMR-LS provide a standard model for use in assessing the pyrethroid resistance in field-collected strains collected from three dengue hotspots: the Taman Seri Bayu (TSB), the Flat Camar (FC), and the Taman Dahlia (TD). Two established methods for determining the resistance mechanisms of the pyrethroid are the quantification of detoxification enzymes via enzyme microassay and the nucleotide sequencing of the domain 2 region from segment 1 to 6 via classical polymerase chain reaction (PCR) amplification-were employed. Enzyme activities in IMR-LS served as the resistance threshold reference, providing a significant standard for comparison with IMR-PSS and other field-collected strains. The amino acids in the domain 2 region of voltage-gated sodium channel (Vgsc) of IMR-LS were served as the reference for detection of any changes of the knockdown resistance (kdr) alleles in IMR-PSS and field-collected strains. Studies clearly indicated that the IMR-LS was highly susceptible to insecticides, whilst the IMR-PSS was highly resistant to pyrethroids and conferred with two resistance mechanisms: the elevated oxidase enzyme activity and the altered target-site mutations. Mutations of V1023G alone, and the combination mutations of V1023G with S996P in IMR-PSS, as well as the in field-collected Aedes aegypti strain, indicate the spread of the (kdr) gene in Aedes aegypti, particularly in dengue-endemic areas in Malaysia.

SUBMITTER: Rasli R

PROVIDER: S-EPMC6023340 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Publications

Susceptibility Status and Resistance Mechanisms in Permethrin-Selected, Laboratory Susceptible and Field-Collected Aedes aegypti from Malaysia.

Rasli Rosilawati R Lee Han Lim HL Wasi Ahmad Nazni N Fikri Siti Futri Farahininajua SFF Ali Roziah R Muhamed Khairul Asuad KA Hadi Azahari Abdul AA Liu Qi-Yong QY Meng Feng Xia FX

Insects 20180418 2

This study is intended to provide a comprehensive characterization of the resistance mechanisms in the permethrin-selected (IMR-PSS) and laboratory susceptible (IMR-LS) <i>Aedes aegypti</i> strain from Malaysia. Both IMR-PSS and IMR-LS provide a standard model for use in assessing the pyrethroid resistance in field-collected strains collected from three dengue hotspots: the Taman Seri Bayu (TSB), the Flat Camar (FC), and the Taman Dahlia (TD). Two established methods for determining the resistan ...[more]

PMID: 29670071

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Project description:Background & objectivesWolbachia-based vector control strategies have been proposed as a mean to augment the existing measures for controlling dengue vector. Prior to utilizing Wolbachia in novel vector control strategies, it is crucial to understand the Wolbachia-mosquito interactions. Many studies have only focused on the prevalence of Wolbachia in female Aedes albopictus with lack of attention on Wolbachia infection on the male Ae. albopictus which also affects the effective expression of Wolbachia induced- cytoplasmic incompatibility (CI). In this study, field surveys were conducted to screen for the infection status of Wolbachia in female and male Ae. albopictus from various habitats including housing areas, islands and seashore.MethodsAdult Ae. albopictus (n=104) were collected using human landing catches and hand aspirator. Standard ovitraps were also set in the selected areas for five days and the larvae were identified to species level. All the collected Ae. albopictus were screened for the presence of Wolbachia using multiplex polymerase chain reaction (PCR) and gene sequencing of Wolbachia surface protein (wsp) gene.ResultsA 100 per cent positivity of Wolbachia infection was observed for individual Ae. albopictus screened. For pooled mosquitoes, 73 of the 76 pools (female) and 83 of the 87 pools (male) were positive with Wolbachia infection. The wsp gene sequence of the Wolbachia strain isolated from individual and pooled mosquitoes showed a 100 per cent homology with Wolbachia sp. of Ae. albopictus isolated from various geographical regions. Phylogenetic analysis based on wsp gene fragments showed that the isolates were clustered into groups A and B, respectively.Interpretation & conclusionsThe results indicated that Wolbachia infection was widespread in Ae. albopictus population both in female and male Ae. albopictus. All the infected females were superinfected with both A and B strains while the infected males showed a combination of superinfection of A and B strains and single infection of B strain.

Project description:The survival of adult female Aedes mosquitoes is a critical component of their ability to transmit pathogens such as dengue viruses. One of the principal determinants of Aedes survival is temperature, which has been associated with seasonal changes in Aedes populations and limits their geographical distribution. The effects of temperature and other sources of mortality have been studied in the field, often via mark-release-recapture experiments, and under controlled conditions in the laboratory. Survival results differ and reconciling predictions between the two settings has been hindered by variable measurements from different experimental protocols, lack of precision in measuring survival of free-ranging mosquitoes, and uncertainty about the role of age-dependent mortality in the field.Here we apply generalised additive models to data from 351 published adult Ae. aegypti and Ae. albopictus survival experiments in the laboratory to create survival models for each species across their range of viable temperatures. These models are then adjusted to estimate survival at different temperatures in the field using data from 59 Ae. aegypti and Ae. albopictus field survivorship experiments. The uncertainty at each stage of the modelling process is propagated through to provide confidence intervals around our predictions.Our results indicate that adult Ae. albopictus has higher survival than Ae. aegypti in the laboratory and field, however, Ae. aegypti can tolerate a wider range of temperatures. A full breakdown of survival by age and temperature is given for both species. The differences between laboratory and field models also give insight into the relative contributions to mortality from temperature, other environmental factors, and senescence and over what ranges these factors can be important.Our results support the importance of producing site-specific mosquito survival estimates. By including fluctuating temperature regimes, our models provide insight into seasonal patterns of Ae. aegypti and Ae. albopictus population dynamics that may be relevant to seasonal changes in dengue virus transmission. Our models can be integrated with Aedes and dengue modelling efforts to guide and evaluate vector control, better map the distribution of disease and produce early warning systems for dengue epidemics.

Project description:Aedes aegypti poses a serious risk to human health due to its wide global distribution, high vector competence for several arboviruses, frequent human biting, and ability to thrive in urban environments. Pyrethroid insecticides remain the primary means of controlling adult A. aegypti populations during disease outbreaks. As a result of decades of use, pyrethroid resistance is a global problem. Cytochrome P450 monooxygenase (CYP)-mediated detoxification is one of the primary mechanisms of pyrethroid resistance. However, the specific CYP(s) responsible for resistance have not been unequivocally determined. We introgressed the resistance alleles from the resistant A. aegypti strain, Singapore (SP), into the genetic background of the susceptible ROCK strain. The resulting strain (CKR) was congenic to ROCK. Our primary goal was to determine which CYPs in SP are linked to resistance. To do this, we first determined which CYPs overexpressed in SP are also overexpressed in CKR, with the assumption that only the CYPs linked to resistance will be overexpressed in CKR relative to ROCK. Next, we determined whether any of the overexpressed CYPs were genetically linked to resistance (cis-regulated) or not (trans-regulated). We found that CYP6BB2, CYP6Z8, CYP9M5 and CYP9M6 were overexpressed in SP as well as in CKR. Based on the genomic sequences and polymorphisms of five single copy CYPs (CYP4C50, 6BB2, 6F2, 6F3 and 6Z8) in each strain, none of these genes were linked to resistance, except for CYP6BB2, which was partially linked to the resistance locus. Hence, overexpression of these four CYPs is due to a trans-regulatory factor(s). Knowledge on the specific CYPs and their regulators involved in resistance is critical for resistance management strategies because it aids in the development of new control chemicals, provides information on potential environmental modulators of resistance, and allows for the detection of resistance markers before resistance becomes fixed in the population.

Dataset Information

Susceptibility Status and Resistance Mechanisms in Permethrin-Selected, Laboratory Susceptible and Field-Collected Aedes aegypti from Malaysia.

Publications

Susceptibility Status and Resistance Mechanisms in Permethrin-Selected, Laboratory Susceptible and Field-Collected Aedes aegypti from Malaysia.

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