Identification and detection of a novel point mutation in the Chitin Synthase gene of Culex pipiens associated with diflubenzuron resistance.


Journal

PLoS neglected tropical diseases
ISSN: 1935-2735
Titre abrégé: PLoS Negl Trop Dis
Pays: United States
ID NLM: 101291488

Informations de publication

Date de publication:
05 2020
Historique:
received: 13 12 2019
accepted: 09 04 2020
revised: 13 05 2020
pubmed: 2 5 2020
medline: 22 7 2020
entrez: 2 5 2020
Statut: epublish

Résumé

Diflubenzuron (DFB) is one of the most used insecticides in mosquito larval control including that of Culex pipiens, the proven vector of the recent West Nile Virus epidemics in Europe. Two mutations (I1043L and I1043M) in the chitin synthase (CHS) putative binding site of DFB have been previously reported in Cx. pipiens from Italy and associated with high levels of resistance against this larvicide. Here we report the identification of a third mutation at the same I1043 position of the CHS gene resulting in the substitution of Isoleucine to Phenylalanine (I1043F). This mutation has also been found in agricultural pests and has been functionally validated with genome editing in Drosophila, showing to confer striking levels (>15,000 fold) of DFB resistance. The frequency of the I1043F mutation was found to be substantially higher in Cx. pipiens mosquitoes surviving DFB doses largely exceeding the recommended field dose, raising concerns about the future efficient use of this insecticide. We monitored the presence and frequency of DFB mutations in Cx. pipiens mosquitoes from several Mediterranean countries, including Italy, France, Greece, Portugal and Israel. Among the Cx. pipiens populations collected in Northern Italy all but one had at least one of the three DFB mutations at allele frequencies reaching 93.3% for the I1043M, 64.8% for the I1043L and 10% for the I1043F. The newly reported I1043F mutation was also identified in two heterozygote individuals from France (4.2% allelic frequency). In contrast to Italy and France, no DFB resistant mutations were identified in the Cx. pipiens mosquitoes sampled from Greece, Portugal and Israel. The findings of our study are of major concern for mosquito control programs in Europe, that rely on the use of a limited number of available larvicides, and highlight the necessity for the development of appropriate Insecticide Resistance Management (IRM) programs, to ensure the sustainable use of DFB.

Sections du résumé

BACKGROUND
Diflubenzuron (DFB) is one of the most used insecticides in mosquito larval control including that of Culex pipiens, the proven vector of the recent West Nile Virus epidemics in Europe. Two mutations (I1043L and I1043M) in the chitin synthase (CHS) putative binding site of DFB have been previously reported in Cx. pipiens from Italy and associated with high levels of resistance against this larvicide.
METHODOLOGY/PRINCIPAL FINDINGS
Here we report the identification of a third mutation at the same I1043 position of the CHS gene resulting in the substitution of Isoleucine to Phenylalanine (I1043F). This mutation has also been found in agricultural pests and has been functionally validated with genome editing in Drosophila, showing to confer striking levels (>15,000 fold) of DFB resistance. The frequency of the I1043F mutation was found to be substantially higher in Cx. pipiens mosquitoes surviving DFB doses largely exceeding the recommended field dose, raising concerns about the future efficient use of this insecticide. We monitored the presence and frequency of DFB mutations in Cx. pipiens mosquitoes from several Mediterranean countries, including Italy, France, Greece, Portugal and Israel. Among the Cx. pipiens populations collected in Northern Italy all but one had at least one of the three DFB mutations at allele frequencies reaching 93.3% for the I1043M, 64.8% for the I1043L and 10% for the I1043F. The newly reported I1043F mutation was also identified in two heterozygote individuals from France (4.2% allelic frequency). In contrast to Italy and France, no DFB resistant mutations were identified in the Cx. pipiens mosquitoes sampled from Greece, Portugal and Israel.
CONCLUSIONS/SIGNIFICANCE
The findings of our study are of major concern for mosquito control programs in Europe, that rely on the use of a limited number of available larvicides, and highlight the necessity for the development of appropriate Insecticide Resistance Management (IRM) programs, to ensure the sustainable use of DFB.

Identifiants

pubmed: 32357192
doi: 10.1371/journal.pntd.0008284
pii: PNTD-D-19-02006
pmc: PMC7219787
doi:

Substances chimiques

Insect Proteins 0
Insecticides 0
Chitin Synthase EC 2.4.1.16
Diflubenzuron J76U6ZSI8D

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e0008284

Déclaration de conflit d'intérêts

he authors have declared that no competing interests exist.

Références

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Auteurs

Emmanouil A Fotakis (EA)

Department of Crop Science, Pesticide Science Lab, Agricultural University of Athens, Athens, Greece.
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Heraklion, Crete, Greece.

Valentina Mastrantonio (V)

Department of Environmental Biology, Sapienza University of Rome, Rome, Italy.

Linda Grigoraki (L)

Vector Biology Dept, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.

Daniele Porretta (D)

Department of Environmental Biology, Sapienza University of Rome, Rome, Italy.

Arianna Puggioli (A)

Medical and Veterinary Entomology Dept., Centro Agricoltura Ambiente "G. Nicoli", Bologna, Italy.

Alexandra Chaskopoulou (A)

European Biological Control Laboratory, USDA-ARS, Thessaloniki, Greece.

Hugo Osório (H)

National Institute of Health Doctor Ricardo Jorge, Centro de Estudos de Vetores e Doenças Infeciosas Doutor Francisco Cambournac, Avenida da Liberdade, Águas de Moura, Portugal.

Mylène Weill (M)

Institut des Sciences de l'Evolution (CNRS-Université de Montpellier-IRD-EPHE), Montpellier, France.

Romeo Bellini (R)

Medical and Veterinary Entomology Dept., Centro Agricoltura Ambiente "G. Nicoli", Bologna, Italy.

Sandra Urbanelli (S)

Department of Environmental Biology, Sapienza University of Rome, Rome, Italy.

John Vontas (J)

Department of Crop Science, Pesticide Science Lab, Agricultural University of Athens, Athens, Greece.
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Heraklion, Crete, Greece.

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