Multiple insecticide resistance in Culex quinquefasciatus populations from Guadeloupe (French West Indies) and associated mechanisms.

West Nile (WN) virus has been detected in Guadeloupe since 2002. Even if no WN human cases have been detected so far, mosquitoes from Culex genus especially Culex quinquefasciatus are recognized as potential WN vectors in Guadeloupe. To evaluate the impact of local vector control activities on this mosquito species we assessed the resistance levels of Cx. quinquefasciatus populations from three different sites from Guadeloupe (Abymes, Saint François and Gourbeyre) to malathion, temephos and deltamethrin. In addition, the frequencies of the L1014F kdr and the G119S ace-1 mutations were established in Cx. quinquefasciatus populations, as well as the constitutive expressions of five cytochrome P450 genes. Mosquito populations tested displayed high resistance to deltamethrin, moderate resistance to malathion (Abymes, Gourbeyre) and low resistance to temephos (Abymes et Gourbeyre). Molecular analyses revealed high frequencies of both L1014F kdr and G119S ace-1 mutations in Cx. quinquefasciatus populations, as well as overexpression of cytochrome P450 genes CYP9J45, CYP9J40 and CYP6AA7. Finally, deltamethrin resistance and knock-down rates were strongly correlated with the frequency of the resistant kdr and ace-1 alleles, as well as with CYP9J40 overexpression. These results should be taken into account to refine the current vector control strategies to prevent the appearance of Cx. quinquefasciatus-borne diseases in Guadeloupe.

Levels of insecticide resistance to deltamethrin, malathion, and temephos, and associated mechanisms in Aedes aegypti mosquitoes from the Guadeloupe and Saint Martin islands (French West Indies).

BACKGROUND: In the Guadeloupe and Saint Martin islands, Aedes aegypti mosquitoes are the only recognized vectors of dengue, chikungunya, and Zika viruses. For around 40 years, malathion was used as a mosquito adulticide and temephos as a larvicide. Since the European Union banned the use of these two insecticide molecules in the first decade of the 21st century, deltamethrin and Bacillus thuringiensis var. israelensis are the remaining adulticide and larvicide, respectively, used in Guadeloupe. In order to improve the management of vector control activities in Guadeloupe and Saint Martin, we investigated Ae. aegypti resistance to and mechanisms associated with deltamethrin, malathion, and temephos. METHODS: Ae. aegypti mosquitoes were collected from six different localities of Guadeloupe and Saint Martin. Larvae were used for malathion and temephos bioassays, and adult mosquitoes for deltamethrin bioassays, following World Health Organization recommendations. Knockdown resistance (Kdr) genotyping for V1016I and F1534C mutations, and expression levels of eight enzymes involved in detoxification mechanisms were examined in comparison with the susceptible reference Bora Bora strain. RESULTS: Resistance ratios (RR50) calculated for Ae. aegypti larvae showed high resistance levels to temephos (from 8.9 to 33.1-fold) and low resistance levels to malathion (from 1.7 to 4.4-fold). Adult females displayed moderate resistance levels to deltamethrin regarding the time necessary to affect 50% of individuals, varying from 8.0 to 28.1-fold. Molecular investigations on adult mosquitoes showed high resistant allele frequencies for V1016I and F1534C (from 85 to 96% and from 90 to 98%, respectively), as well as an overexpression of the glutathione S-transferase gene, GSTe2, the carboxylesterase CCEae3a, and the cytochrome genes 014614, CYP6BB2, CYP6M11, and CYP9J23. CONCLUSIONS: Ae. aegypti populations from Guadeloupe and Saint Martin exhibit multiple resistance to organophosphates (temephos and malathion), and pyrethroids (deltamethrin). The mechanisms associated with these resistance patterns show strong frequencies of F1534C and V1016I Kdr mutations, and an over-expression of CCEae3a, GSTe2, and four cytochrome P450 genes (014614, CYP9J23, CYP6M11, CYP6BB2). These results will form the baseline for a deeper understanding of the insecticide resistance levels and associated mechanisms of Ae. aegypti populations and will be used to improve vector control strategies in Guadeloupe and Saint Martin.

Parity and longevity of Aedes aegypti according to temperatures in controlled conditions and consequences on dengue transmission risks.

BACKGROUND: In Guadeloupe, Aedes aegypti mosquitoes are the only vectors of dengue and chikungunya viruses. For both diseases, vector control is the only tool for preventing epidemics since no vaccine or specific treatment is available. However, to efficiently implement control of mosquitoes vectors, a reliable estimation of the transmission risks is necessary. To become infective an Ae. aegypti female must ingest the virus during a blood meal and will not be able to transmit the virus during another blood-meal until the extrinsic incubation period is completed. Consequently the aged females will carry more infectious risks. The objectives of the present study were to estimate under controlled conditions the expectation of infective life for females and thus the transmission risks in relation with their reproductive cycle and parity status. METHODOLOGY/PRINCIPAL FINDINGS: Larvae of Ae. aegypti were collected in central Guadeloupe and breed under laboratory conditions until adult emergence. The experiments were performed at constant temperatures (± 1.5°C) of 24°C, 27°C and 30°C on adults females from first generation (F1). Females were kept and fed individually and records of blood-feeding, egg-laying and survival were done daily. Some females were dissected at different physiological stages to observe the ovaries development. The data were analyzed to follow the evolution of parity rates, the number of gonotrophic cycles, the fecundity and to study the mean expectation of life and the mean expectation of infective life for Ae. aegypti females according to temperatures. The expectation of life varies with the parity rates and according to the temperatures, with durations from about 10 days at low parity rates at the higher temperature to an optimal duration of about 35 days when 70% of females are parous at 27°C. Infective life expectancy was found highly variable in the lower parous rates and again the optimal durations were found when more than 50% of females are parous for the mean temperatures of 27°C and 30°C. CONCLUSION: Parity rates can be determined for field collected females and could be a good proxy of the expectation of infective life according to temperatures. However, for the same parity rates, the estimation of infective life expectation is very different between Ae. aegypti and Anopheles gambiae mosquitoes. Correlation of field parity rates with transmission risks requires absolutely to be based on Ae. aegypti models, since available Anopheles sp. models underestimate greatly the females longevity.

Increasing Role of Roof Gutters as Aedes aegypti (Diptera: Culicidae) Breeding Sites in Guadeloupe (French West Indies) and Consequences on Dengue Transmission and Vector Control.

During the past ten years, the islands of Guadeloupe (French West Indies) are facing dengue epidemics with increasing numbers of cases and fatal occurrences. The vector Aedes aegypti is submitted to intensive control, with little effect on mosquito populations. The hypothesis that important Ae. aegypti breeding sites are not controlled is investigated herein. For that purpose, the roof gutters of 123 houses were systematically investigated, and the percentage of gutters positive for Ae. aegypti varied from 17.2% to 37.5%, from humid to dry locations. In the dryer location, most of houses had no other breeding sites. The results show that roof gutters are becoming the most important Ae. aegypti breeding sites in some locations in Guadeloupe, with consequences on dengue transmission and vector control.

High susceptibility to Chikungunya virus of Aedes aegypti from the French West Indies and French Guiana.

OBJECTIVES: To estimate the vector competence of Aedes aegypti populations sampled from distinct anthropogenic environments in French Guiana, Guadeloupe and Martinique for the strain CHIKV 06.21. METHODS: F(1)/F(2) females were orally infected at titres of 10(6) and 10(7.5) pfu/ml in blood-meals. Disseminated infection rates (DIR) of mosquitoes were estimated using indirect fluorescent antibody assay on heads' squashes, 7 or 14 days post-infection (pi). RESULTS: At a titre of 10(7.5) pfu/ml, DIR ranged from 88.9% to 100.0% and were not significantly different whether assessed at day 7 or 14 pi. At a titre of 10(6) pfu/ml, DIR observed 7 days pi ranged from 37.6 to 62.0%. CONCLUSIONS: Ae. aegypti from French Guiana and French West Indies are highly competent to transmit CHIKV. An evaluation of DIR 7 days rather than 14 days pi is adequate to estimate vector competence. The titre of 10(6) pfu/ml allows us to distinguish Ae. aegypti populations originating from distinct environments (dense or diffuse housing) by their vector competence. This assessment is a prerequisite to better evaluate the potential risk of Chikungunya outbreaks once the virus is introduced from endemic regions.