Diversity and frequency of kdr mutations within Anopheles sinensis populations from Guangxi, China

Yang, Chan; Xu, Feng; Huang, Zhuo; Li, Mei; Qiu, Xinghui

doi:10.1186/s12936-016-1467-3

Cited by 16 publications

(33 citation statements)

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“…Both H3 and H17 were distributed in all the nine populations tested, and shared the type-2 intron. Similar to the status of AS - VGSC [ 12 ], the high polymorphism of AsRDL may be partly explained by their large population size, wide distribution range of An. sinensis [ 36 ], diverse natural landscapes and different local insecticide selective pressure in Guangxi.…”

Section: Discussionmentioning

confidence: 94%

“…The sampling sites were part of the malaria vector monitoring sites that were set up partly because there were imported malaria cases observed since 2010 in these regions. A brief description of the sampling sites was provided previously [ 12 ].…”

Section: Methodsmentioning

confidence: 99%

“…Although the distribution and frequency of insecticide resistance-conferring mutations in the acetylcholinesterase and voltage gated sodium channel were determined in An. sinensis collected extensively across Guangxi in previous studies [ 11 , 12 ], other questions related to insecticide resistance remain to be answered.…”

Section: Introductionmentioning

confidence: 99%

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RDL mutations predict multiple insecticide resistance in Anopheles sinensis in Guangxi, China

Yang

Huang

Mei

et al. 2017

Malar J

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Background Anopheles sinensis is a major vector of malaria in China. The gamma-aminobutyric acid (GABA)-gated chloride channel, encoded by the RDL (Resistant to dieldrin) gene, is the important target for insecticides of widely varied structures. The use of various insecticides in agriculture and vector control has inevitably led to the development of insecticide resistance, which may reduce the control effectiveness. Therefore, it is important to investigate the presence and distribution frequency of the resistance related mutation(s) in An. sinensis RDL to predict resistance to both the withdrawn cyclodienes (e.g. dieldrin) and currently used insecticides, such as fipronil.MethodsTwo hundred and forty adults of An. sinensis collected from nine locations across Guangxi Zhuang Autonomous Region were used. Two fragments of An. sinensis RDL (AsRDL) gene, covering the putative insecticide resistance related sites, were sequenced respectively. The haplotypes of each individual were reconstructed by the PHASE2.1 software, and confirmed by clone sequencing. The phylogenetic tree was built using maximum-likelihood and Bayesian inference methods. Genealogical relations among different haplotypes were also analysed using Network 5.0.ResultsThe coding region of AsRDL gene was 1674 bp long, encoding a protein of 557 amino acids. AsRDL had 98.0% amino acid identity to that from Anopheles funestus, and shared common structural features of Cys-loop ligand-gated ion channels. Three resistance-related amino acid substitutions (A296S, V327I and T345S) were detected in all the nine populations of An. sinensis in Guangxi, with the 296S mutation being the most abundant (77–100%), followed by 345S (22–47%) and 327I (8–60%). 38 AsRDL haplotypes were identified from 240 individuals at frequencies ranging from 0.2 to 34.8%. Genealogical analysis suggested multiple origins of the 345S mutation in AsRDL.ConclusionsThe near fixation of the 296S mutation and the occurrence of the 327I and 345S mutations in addition to 296S, in all the nine tested An. sinensis populations in Guangxi, strongly indicate a risk of multiple insecticide resistance. The haplotype diversity plus genetic heterogeneities in the geographical distribution, and multiple origins of AsRDL alleles call for a location-customized strategy for monitoring and management of insecticide resistance.

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Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

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RDL mutations predict multiple insecticide resistance in Anopheles sinensis in Guangxi, China

Yang

Huang

Mei

et al. 2017

Malar J

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“…In addition to 1014F, other two mutations (1014S and 1014C) were detected in An. sinensis from Asia, including China [16, 18–20]. Interestingly, kdr frequencies in samples from western Guangxi of China were low, while those in samples from northeast Guangxi were high [20].…”

Section: Introductionmentioning

confidence: 99%

Target-site mutations (AChE-G119S and kdr) in Guangxi Anopheles sinensis populations along the China-Vietnam border

Yang

Liu

et al. 2019

Parasites Vectors

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BackgroundIn South Asia, the epidemiology of malaria is complex, and transmission mainly occurs in remote areas near international borders. Vector control has been implemented as a key strategy in malaria prevention for decades. A rising threat to the efficacy of vector control efforts is the development of insecticide resistance, thus it is important to monitor the type and frequency of insecticide resistant alleles in the disease vectors such as An. sinensis along the China-Vietnam border. Such information is needed to synthesize effective malaria vector control strategies.MethodsA total of 208 adults of An. sinensis, collected from seven sites in southwest Guangxi along the China-Vietnam border, were inspected for the resistance-conferring G119S mutation in acetylcholinesterase (AChE) by PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism) and kdr mutations in the voltage-gated sodium channel (VGSC) by sequencing. In addition, the evolutionary origin of An. sinensis vgsc gene haplotypes was analyzed using Network 5.0.ResultsThe frequencies of mutant 119S of AChE were between 0.61–0.85 in the seven An. sinensis populations. No susceptible homozygote (119GG) was detected in three of the seven sites (DXEC, LZSK and FCGDX). Very low frequencies of kdr (0.00–0.01) were detected in the seven populations, with most individuals being susceptible homozygote (1014LL). The 1014F mutation was detected only in the southeast part (FCGDX) at a low frequency of 0.03. The 1014S mutation was distributed in six of the seven populations with frequencies ranging from 0.04 to 0.08, but absent in JXXW. Diverse haplotypes of 1014L and 1014S were found in An. sinensis along the China-Vietnam border, while only one 1014F haplotype was detected in this study. Consistent with a previous report, resistant 1014S haplotypes did not have a single origin.ConclusionsThe G119S mutation of AChE was present at high frequencies (0.61–0.85) in the An. sinensis populations along the China-Vietnam border, suggesting that the vector control authorities should be cautious when considering carbamates and organophosphates as chemicals for vector control. The low frequencies (0.00–0.11) of kdr in these populations suggest that pyrethroids remain suitable for use against An. sinensis in these regions.

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“…The evolution of pyrethroid resistance occurs through complicated mechanisms typically involving interactions between multiple categories of gene . Considerable progress has been made in the characterization and functional analyses of detoxification‐related genes and mutations in the target‐site membrane protein Kdr …”

Section: Introductionmentioning

confidence: 99%

Mitochondrial genes associated with pyrethroid resistance revealed by mitochondrial genome and transcriptome analyses in the malaria vector Anopheles sinensis (Diptera: Culicidae)

Ding

Yan

et al. 2019

Pest Management Science

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BACKGROUND: Insecticides are still the main method of mosquito control, but mosquito resistance presents a large obstacle. The function of mitochondrial genes in the evolution of insecticide resistance is still poorly understood. Pyrethroid is the most commonly used insecticide, and Anopheles sinensis is an important malaria vector in China and Southeast Asia. In this study, we investigated the mitochondrial genes associated with pyrethroid resistance through their genetic and expression variation based on analyses of transcriptomes and 36 individuals with resequencing in three geographical populations in China. RESULTS:The nucleotide diversity (Pi) in 18 resistant individuals was much lower than that in 18 susceptible individuals, which suggests that some sites experienced purifying selection subject to pyrethroid stress. Ka/Ks and amino acid analyses showed that ND4 experienced positive selection and had 23 amino acid mutations due to pyrethroid stress. These mutations might change the ND4 structure and function and thus alter the efficiency of the respiratory chain. ND5 was significantly upregulated, and ATP8 was significantly downregulated in these three pyrethroid resistant populations, which suggests that these two genes function in the production and maintenance of pyrethroid resistance. There are differences in mitochondrial genes involved in pyrethroid resistance among these three populations.CONCLUSION: This is the first study to reveal the association of mitochondrial genes in the evolution of insecticide resistance through amino acid mutation and expression patterns and can help us further understand insecticide resistance mechanisms.

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Diversity and frequency of kdr mutations within Anopheles sinensis populations from Guangxi, China

Cited by 16 publications

References 33 publications

RDL mutations predict multiple insecticide resistance in Anopheles sinensis in Guangxi, China

RDL mutations predict multiple insecticide resistance in Anopheles sinensis in Guangxi, China

Target-site mutations (AChE-G119S and kdr) in Guangxi Anopheles sinensis populations along the China-Vietnam border

Mitochondrial genes associated with pyrethroid resistance revealed by mitochondrial genome and transcriptome analyses in the malaria vector Anopheles sinensis (Diptera: Culicidae)

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