Molecular characterization of DDT resistance in Anopheles gambiae from Benin

Particle and Fibre Toxicology

Vincent Corbel⁶  Hilary Ranson¹  Martin Akogbéto⁴  Sylvie Cornelie²  Rodolphe Poupardin¹  Christopher M Jones³  Fiacre R Agossa⁴  Innocent Djègbè⁵ 

[1] Vector Group, Liverpool School of Tropical Medicine (LSTM), Liverpool L3 5QA, UK;Centre de Recherche Entomologique de Cotonou (CREC), 06 BP 2604, Cotonou, Bénin;Insect Migration & Spatial Ecology; Group AgroEcology Rothamsted, Research Harpenden, Hertfordshire AL5 2JQ, UK;Département de Zoologie, Faculté des Sciences et Techniques (FAST), Université d’Abomey Calavi (UAC), BP 526 Cotonou, Bénin;Ecole Normale Supérieure de Natitingou, Université de Parakou, BP 123 Parakou, Benin;Department of Entomology, Faculty of Agriculture, Kasetsart University, Bangkok 10900, Thailand

关键词: Metabolic enzymes;    Vector control;    Kdr mutation;    qPCR;    Insecticide resistance;    An. gambiae;   

Others  :  1150672
DOI  :  10.1186/1756-3305-7-409

 received in 2014-05-02, accepted in 2014-08-20,  发布年份 2014

【 摘 要 】

Background

Insecticide resistance in the mosquito vector is the one of the main obstacles against effective malaria control. In order to implement insecticide resistance management strategies, it is important to understand the genetic factors involved. In this context, we investigated the molecular basis of DDT resistance in the main malaria vector from Benin.

Methods

Anopheles gambiae mosquitoes were collected from four sites across Benin and identified to species/molecular form. Mosquitoes from Cotonou (M-form), Tori-Bossito (S-form) and Bohicon (S-form) were exposed to DDT 4% at a range of exposure times (30 min to 300 min). Another batch of mosquitoes from Cotonou and Malanville were exposed to DDT for 1 hour and the survivors 48 hours post exposure were used to quantify metabolic gene expression. Quantitative PCR assays were used to quantify mRNA levels of metabolic enzymes: GSTE2, GSTD3, CYP6P3 and CYP6M2. Expression (fold-change) was calculated using the ∆∆Ct method and compared to susceptible strains. Detection of target-site mutations (L1014F, L1014S and N1575Y) was performed using allelic discrimination TaqMan assays.

Results

DDT resistance was extremely high in all populations, regardless of molecular form, with no observed mortality after 300 min exposure. In both DDT-survivors and non-exposed mosquitoes, GSTE2 and GSTD3 were over-expressed in the M form at 4.4-fold and 3.5-fold in Cotonou and 1.5-fold and 2.5-fold in Malanville respectively, when compared to the susceptible strain. The CYP6M2 and CYP6P3 were over-expressed at 4.6-fold and 3.8-fold in Cotonou and 1.2-fold and 2.5-fold in Malanville respectively. In contrast, no differences in GSTE2 and CYP6M2 were observed between S form mosquitoes from Tori-Bossito and Bohicon compared to susceptible strain. The 1014 F allele was fixed in the S-form and at high frequency in the M-form (0.7-0.914). The frequency of 1575Y allele was 0.29-0.36 in the S-form and nil in the M-form. The 1014S allele was detected in the S form of An. gambiae in a 1014 F/1014S heterozygous specimen.

Conclusion

Our results show that the kdr 1014 F, 1014S and 1575Y alleles are widespread in Benin and the expression of two candidate metabolic markers (GSTE2 and CYP6M2) are over-expressed specifically in the M-form.

【 授权许可】

   
2014 Djègbè et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Hemingway J, Hawkes NJ, McCarroll L, Ranson H: The molecular basis of insecticide resistance in mosquitoes. Insect Biochem Mol Biol 2004, 34(7):653-665.
• [2]Santolamazza F, Calzetta M, Etang J, Barrese E, Dia I, Caccone A, Donnelly MJ, Petrarca V, Simard F, Pinto J, Torre A d: Distribution of knock-down resistance mutations in Anopheles gambiae molecular forms in west and west-central Africa. Malar J 2008, 7:74. BioMed Central Full Text
• [3]Lynd A, Ranson H, McCall PJ, Randle NP, Black WC, Walker ED, Donnelly MJ: A simplified high-throughput method for pyrethroid knock-down resistance (kdr) detection in Anopheles gambiae. Malar J 2005, 4(1):16. BioMed Central Full Text
• [4]Ranson H, N’Guessan R, Lines J, Moiroux N, Nkuni Z, Corbel V: Pyrethroid resistance in African anopheline mosquitoes: what are the implications for malaria control? Trends Parasitol 2011, 27(2):91-98.
• [5]Donnelly MJ, Corbel V, Weetman D, Wilding CS, Williamson MS, Black WC IV: Does kdr genotype predict insecticide-resistance phenotype in mosquitoes? Trends Parasitol 2009, 25(5):213-219.
• [6]Jones CM, Liyanapathirana M, Agossa FR, Weetman D, Ranson H, Donnelly MJ, Wilding CS: Footprints of positive selection associated with a mutation (N1575Y) in the voltage-gated sodium channel of Anopheles gambiae. Proc Natl Acad Sci 2012, 109(17):6614-6619.
• [7]Djouaka RF, Bakare AA, Coulibaly ON, Akogbeto MC, Ranson H, Hemingway J, Strode C: Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria. BMC Genomics 2008, 9:538. BioMed Central Full Text
• [8]Corbel V, N'Guessan R, Brengues C, Chandre F, Djogbenou L, Martin T, Akogbeto M, Hougard JM, Rowland M: Multiple insecticide resistance mechanisms in Anopheles gambiae and Culex quinquefasciatus from Benin. West Africa. Acta Trop 2007, 101(3):207-216.
• [9]Etang J, Manga L, Chandre F, Guillet P, Fondjo E, Mimpfoundi R, Toto JC, Fontenille D: Insecticide susceptibility status of Anopheles gambiae s.l. (Diptera: Culicidae) in the Republic of Cameroon. J Med Entomol 2003, 40(4):491-497.
• [10]Müller P, Warr E, Stevenson BJ, Pignatelli PM, Morgan JC, Steven A, Yawson AE, Mitchell SN, Ranson H, Hemingway J: Field-caught permethrin-resistant Anopheles gambiae overexpress CYP6P3, a P450 that metabolises pyrethroids. PLoS Genet 2008, 4(11):1000286.
• [11]Stevenson BJ, Bibby J, Pignatelli P, Muangnoicharoen S, O'Neill PM, Lian LY, Müller P, Nikou D, Steven A, Hemingway J: Cytochrome P450 6 M2 from the malaria vector Anopheles gambiae metabolizes pyrethroids: Sequential metabolism of deltamethrin revealed. Insect Biochem Mol Biol 2011, 41(7):492-502.
• [12]Nikou D, Ranson H, Hemingway J: An adult-specific CYP6 P450 gene is overexpressed in a pyrethroid-resistant strain of the malaria vector, Anopheles gambiae. Gene 2003, 318:91-102.
• [13]Nwane P, Etang J, Chouaibou M, Toto J, Koffi A, Mimpfoundi R, Simard F: Multiple insecticide resistance mechanisms in Anopheles gambiae sl populations from Cameroon, Central Africa. Parasit Vectors 2013, 6:41-54. BioMed Central Full Text
• [14]Jones CM, Haji KA, Khatib BO, Bagi J, Mcha J, Devine GJ, Daley M, Kabula B, Ali AS, Majambere S: The dynamics of pyrethroid resistance in Anopheles arabiensis from Zanzibar and an assessment of the underlying genetic basis. Parasit Vectors 2013, 6:343. BioMed Central Full Text
• [15]Mitchell SN, Stevenson BJ, Müller P, Wilding CS, Egyir-Yawson A, Field SG, Hemingway J, Paine MJI, Ranson H, Donnelly MJ: Identification and validation of a gene causing cross-resistance between insecticide classes in Anopheles gambiae from Ghana. Proc Natl Acad Sci 2012, 109(16):6147-6152.
• [16]Ranson H, Rossiter L, Ortelli F, Jensen B, Wang X, Roth CW, Collins FH, Hemingway J: Identification of a novel class of insect glutathione S-transferases involved in resistance to DDT in the malaria vector Anopheles gambiae. Biochem J 2001, 359:295-304.
• [17]WHO: Launch of the Global Plan for Insecticide Resistance Management in malaria vectors. Geneva: ᅟ; 2012. http://www.who.int/malaria/vector_control/gpirm_event_report_2012.pdf webcite
• [18]Djegbe I, Boussari O, Sidick A, Martin T, Ranson H, Chandre F, Akogbeto M, Corbel V: Dynamics of insecticide resistance in malaria vectors in Benin: first evidence of the presence of L1014S kdr mutation in Anopheles gambiae from West Africa. Malar J 2011, 10:261. BioMed Central Full Text
• [19]N'Guessan R, Corbel V, Akogbeto M, Rowland M: Reduced efficacy of insecticide-treated nets and indoor residual spraying for malaria control in pyrethroid resistance area, Benin. Emerg Infect Dis 2007, 13(2):199-206.
• [20]Corbel V, Akogbeto M, Damien GB, Djenontin A, Chandre F, Rogier C, Moiroux N, Chabi J, Padonou GG, Henry MC: Combination of malaria vector control interventions in pyrethroid resistance area in Benin: a cluster randomised controlled trial. The Lancet Infect Dis 2012, 12(8):617-626.
• [21]Asidi A, N’Guessan R, Akogbeto M, Curtis C, Rowland M: Loss of Household Protection from Use of Insecticide-Treated Nets against Pyrethroid-Resistant Mosquitoes, Benin. Emerg Infect Dis 2012, 18(7):1101.
• [22]WHO: Tests procedures for insecticide resistance monitoring in malaria vector, bio-efficacy and persistence of insecticides on treated surfaces. WHO/CDS/CPC/MAL/98 1998, 12:43.
• [23]Livak KJ: Organization and mapping of a sequence on the Drosophila melanogaster X and Y chromosomes that is transcribed during spermatogenesis. Genetics 1984, 107(4):611-634.
• [24]Bass C, Nikou D, Donnelly M, Williamson M, Ranson H, Ball A, Vontas J, Field L: Detection of knockdown resistance (kdr) mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods. Malar J 2007, 6(1):111. BioMed Central Full Text
• [25]Witzig C, Parry M, Morgan J, Irving H, Steven A, Cuamba N, Kerah-Hinzoumbe C, Ranson H, Wondji C: Genetic mapping identifies a major locus spanning P450 clusters associated with pyrethroid resistance in kdr-free Anopheles arabiensis from Chad. Heredity 2013, 110(4):389-397.
• [26]Schmittgen TD, Livak KJ: Analyzing real-time PCR data by the comparative CT method. Nat Protoc 2008, 3(6):1101-1108.
• [27]Namountougou M, Diabaté A, Etang J, Bass C, Sawadogo SP, Gnankinié O, Baldet T, Martin T, Chandre F, Simard F: First report of the L1014S kdr mutation in wild populations of Anopheles gambiae M and S molecular forms in Burkina Faso (West Africa). Acta Trop 2012, 7(11):48412.
• [28]Nardini L, Christian RN, Coetzer N, Koekemoer LL: DDT and pyrethroid resistance in Anopheles arabiensis from South Africa. Parasit Vectors 2013, 6:229. BioMed Central Full Text
• [29]Badolo A, Traoré A, Jones CM, Sanou A, Flood L, Guelbéogo WM, Ranson H, Sagnon NF: Three years of insecticide resistance monitoring in Anopheles gambiae in Burkina Faso: resistance on the rise? Malar J 2012, 11(1):232. BioMed Central Full Text
• [30]Jones CM, Toé HK, Sanou A, Namountougou M, Hughes A, Diabaté A, Dabiré R, Simard F, Ranson H: Additional selection for insecticide resistance in urban malaria vectors: DDT resistance in Anopheles arabiensis from Bobo-Dioulasso, Burkina Faso. PLoS One 2012, 7(9):45995.
• [31]Prapanthadara L, Hemingway J, Ketterman A: Partial purification and characterization of Glutathione-S-Transferases involved in DDT resistance from the mosquito Anopheles gambiae. Pesticide Biochem Physiol 1993, 47(2):119-133.
• [32]Grant DF, Dietze EC, Hammock BD: Glutathione-S-transferase isozymes in Aedes aegypti: purification, characterization, and isozyme-specific regulation. Insect Biochem 1991, 21(4):421-433.
• [33]Che-Mendoza A, Penilla RP, Rodríguez DA: Insecticide resistance and glutathione S-transferases in mosquitoes: a review. Afr J Biotechnol 2009, 8:8.
• [34]Ding Y, Hawkes N, Meredith J, Eggleston P, Hemingway J, Ranson H: Characterization of the promoters of Epsilon glutathione transferases in the mosquito Anopheles gambiae and their response to oxidative stress. Biochem J 2005, 387:879-888.
• [35]Vontas J, Blass C, Koutsos AC, David JP, Kafatos FC, Louis C, Hemingway J, Christophides GK, Ranson H: Gene expression in insecticide resistant and susceptible Anopheles gambiae strains constitutively or after insecticide exposure. Insect Mol Biol 2005, 14(5):509-521.
• [36]Enayati A, Ranson H, Hemingway J: Insect glutathione transferases and insecticide resistance. Insect Mol Biol 2005, 14(1):3-8.
• [37]David JP, Strode C, Vontas J, Nikou D, Vaughan A, Pignatelli PM, Louis C, Hemingway J, Ranson H: The Anopheles gambiae detoxification chip: a highly specific microarray to study metabolic-based insecticide resistance in malaria vectors. Proc Natl Acad Sci 2005, 102(11):4080-4084.
• [38]Djouaka R, Irving H, Tukur Z, Wondji CS: Exploring mechanisms of multiple insecticide resistance in a population of the malaria vector Anopheles funestus in Benin. PLoS One 2011, 6(11):27760.
• [39]Riveron JM, Yunta C, Ibrahim SS, Djouaka R, Irving H, Menze BD, Ismail HM, Hemingway J, Ranson H, Albert A: A single mutation in the GSTe2 gene allows tracking of metabolically based insecticide resistance in a major malaria vector. Genome Biol 2014, 15(2):27. BioMed Central Full Text
• [40]Mitchell SN, Rigden DJ, Dowd AJ, Lu F, Wilding CS, Weetman D, Dadzie S, Jenkins AM, Regna K, Boko P: Metabolic and target-site mechanisms combine to confer strong DDT resistance in Anopheles gambiae. PLoS One 2014, 9(3):92662.
• [41]della Torre A, Tu Z, Petrarca V: On the distribution and genetic differentiation of Anopheles gambiae ss molecular forms. Insect Biochem Mol Biol 2005, 35(7):755-769.
• [42]Costantini C, Ayala D, Guelbeogo WM, Pombi M, Some CY, Bassole IHN, Ose K, Fotsing JM, Sagnon NF, Fontenille D: Living at the edge: biogeographic patterns of habitat segregation conform to speciation by niche expansion in Anopheles gambiae. BMC Ecol 2009, 9(1):16. BioMed Central Full Text
• [43]Simard F, Ayala D, Kamdem GC, Pombi M, Etouna J, Ose K, Fotsing J-M, Fontenille D, Besansky NJ, Costantini C: Ecological niche partitioning between Anopheles gambiae molecular forms in Cameroon: the ecological side of speciation. BMC Ecol 2009, 9(1):17. BioMed Central Full Text
• [44]Poupardin R, Reynaud S, Strode C, Ranson H, Vontas J, David JP: Cross-induction of detoxification genes by environmental xenobiotics and insecticides in the mosquito Aedes aegypti: Impact on larval tolerance to chemical insecticides. Insect Biochem Molr Biol 2008, 38(5):540-551.
• [45]Djouaka RF, Bakare AA, Bankole HS, Doannio JM, Coulibaly ON, Kossou H, Tamo M, Basene HI, Popoola OK, Akogbeto MC: Does the spillage of petroleum products in Anopheles breeding sites have an impact on the pyrethroid resistance? Malar J 2007, 6:159. BioMed Central Full Text
• [46]Djouaka RF, Bakare AA, Bankole HS, Doannio JM, Kossou H, Akogbeto MC: Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin. Malar J 2007, 6:56. BioMed Central Full Text
• [47]Tene Fossog B, Kopya E, Ndo C, Menze-Djantio B, Costantini C, Njiokou F, Awono-Ambene P, Antonio-Nkondjio C: Water quality and Anopheles gambiae larval tolerance to Pyrethroids in the cities of Douala and Yaoundé (Cameroon). J Trop Med 2012, 2012(1155):429817.
• [48]Yu SJ, Hsu EL: Induction of detoxification enzymes in phytophagous insects: role of insecticide synergists, larval age, and species. Archives Insect Biochem Physiol 1993, 24(1):21-32.
• [49]N'Guessan R, Boko P, Odjo A, Chabi J, Akogbeto M, Rowland M: Control of pyrethroid and DDT-resistant Anopheles gambiae by application of indoor residual spraying or mosquito nets treated with a long-lasting organophosphate insecticide, chlorpyrifos-methyl. Malar J 2010, 9:44. BioMed Central Full Text