CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >
Resistance to deltamethrin and knockdown resistance mutation in Aedes albopictus from Jiangsu Province
Received date: 2021-12-27
Revised date: 2022-03-15
Online published: 2022-09-07
Supported by
National Key R&D Program of China(2020YFC1200105);Jiangsu Provincial Department of Science and Technology(BM2018020)
Objective To investigate the resistance of deltamethrin in Aedes albopictus in Jiangsu Province, and explore the association with knockdown resistance (kdr) mutation, to provide a theoretical basis for scientific control of Ae. albopictus. Methods The diagnostic doses were determined by preparing test bottles containing deltamethrin of 1, 2, 4, 6 or 8 μg/bottle and control bottles without deltamethrin using the CDC bottle bioassay with 10-25 susceptible laboratory-cultured Ae. albopictus in each bottle. Ae. albopictus larvae and pupae were collected from Suining County in Xuzhou City, Huaiyin District in Huai’an City, and Yixing City in Wuxi City, in Jiangsu Province during August and September 2020 and reared to the first filial generation (F1 generation). The mortality rate of Ae. albopictus was calculated by using 10-25 F1 generation female mosquitoes for exposure to a diagnostic dose of deltamethrin in the test bottle and the blank in the control bottle for 30 min to evaluate the resistance level. The DNA was extracted from the mosquitoes after bioassay, and the gene coding region of voltage-gated sodium channel (VGSC) transmembrane structural domains Ⅱ, Ⅲ and Ⅳwere amplified by PCR and sequenced. Bioedit and Seqman software were used for statistical analysis of the kdr mutations, genotypes and mutation frequency. A Chi-square test was used to compare the kdr mutation frequency in the resistant and susceptible phenotype mosquitoes. Results The diagnostic doses of deltamethrin to Ae. albopictus was 4 μg/bottle. A total of 149 Ae. albopictus were collected from Suining County, Huaiyin District and Yixing City. The corrected mean mortality rate of Suining County was (28.2 ± 1.4)%, and the mean mortality rates in Huaiyin District and Yixing City were (64.4 ± 5.5)% and (56.3 ± 17.7)%, respectively. Ae. albopictus collected from all three counties’ (cities and districts) were resistant to deltamethrin. There were 447 DNA sequences obtained, and the sequencing results showed that the kdr mutations existed in the gene coding region at codons 1532 and 1534 of the VGSC structural domain Ⅳ of Ae. albopictus in three counties (cities and districts). Two alleles at codon 1532 were wild-type ATC/I (279/298, 93.6%) and mutant ACC/T (19/298, 6.4%); three genotypes were the wild-type homozygote I/I (133/149, 89.3%), wild/mutant heterozygote I/T (13/149, 8.7%), and mutant homozygote T/T (3/149, 2.0%). Four alleles at codon 1534 were wildtype TTC/F (275/298, 92.3%), mutant TCC/S (5/298, 1.7%), mutant TTA/L (14/298, 4.7%), and mutant TTG/L (4/298, 1.3%); four genotypes were the wild-type homozygote F/F (128/149, 85.9%), wild/mutant heterozygote F/S (5/149, 3.4%), wild/mutant heterozygote F/L (14/149, 9.4%), and mutant homozygote I/I (2/149, 1.3%). Among the 72 resistant mosquitoes phenotyped, 87.5% (63/72) and 86.1% (62/72) were wild-type at codon 1532 and 1534, respectively, and 12.5% (9/72) and 13.9% (10/72) were kdr mutant, respectively. Among the 77 sensitive adults, 90.9% (70/77) and 85.7% (66/77) were wild type at codon 1532 and 1534, respectively, and 9.2% (7/77) and 14.3% (11/77) were kdr mutant, respectively. Chi-square test revealed no significant difference between kdr mutant genotypes at codon 1532 and 1534 with resistant and susceptible phenotypes Ae. albopictus( χ2 = 0.168, 4.111, P > 0.05). Conclusion In Suining County, Huaiyin District and Yixing City of Jiangsu Province, Ae. albopictus mosquitoes were found resistant to deltamethrin. The frequencies of kdr mutations at codon 1532 and 1534 in Ae. albopictus was low. No significant correlation was found between the resistance phenotype and kdr mutations at codon 1532 and 1534.
WANG Guan-xi , LI Ya-shu , LI Yue-yue , CAO Yuan-yuan , YANG Meng-meng , ZHANG Mei-hua , WU Jing-yao , LIANG Cheng , LI Ju-lin , ZHOU Hua-yun , TANG Jian-xia , ZHU Guo-ding . Resistance to deltamethrin and knockdown resistance mutation in Aedes albopictus from Jiangsu Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022 , 40(4) : 468 -474 . DOI: 10.12140/j.issn.1000-7423.2022.04.008
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