Entomological surveys and insecticide susceptibility profile of Aedes aegypti during the dengue outbreak in Sao Tome and Principe in 2022.

BACKGROUND: The first dengue outbreak in Sao Tome and Principe was reported in 2022. Entomological investigations were undertaken to establish the typology of Aedes larval habitats, the distribution of Ae. aegypti and Ae. albopictus, the related entomological risk and the susceptibility profile of Ae. aegypti to insecticides, to provide evidence to inform the outbreak response. METHODOLOGY/PRINCIPAL FINDINGS: Entomological surveys were performed in all seven health districts of Sao Tome and Principe during the dry and rainy seasons in 2022. WHO tube and synergist assays using piperonyl butoxide (PBO) and diethyl maleate (DEM) were carried out, together with genotyping of F1534C/V1016I/V410L mutations in Ae. aegypti. Aedes aegypti and Ae. albopictus were found in all seven health districts of the country with high abundance of Ae. aegypti in the most urbanised district, Agua Grande. Both Aedes species bred mainly in used tyres, discarded tanks and water storage containers. In both survey periods, the Breteau (BI > 50), house (HI > 35%) and container (CI > 20%) indices were higher than the thresholds established by WHO to indicate high potential risk of dengue transmission. The Ae. aegypti sampled were susceptible to all insecticides tested except dichlorodiphenyltrichloroethane (DDT) (9.2% mortality, resistant), bendiocarb (61.4% mortality, resistant) and alpha-cypermethrin (97% mortality, probable resistant). A full recovery was observed in Ae. aegypti resistant to bendiocarb after pre-exposure to synergist PBO. Only one Ae. aegypti specimen was found carrying F1534C mutation. CONCLUSIONS/SIGNIFICANCE: These findings revealed a high potential risk for dengue transmission throughout the year, with the bulk of larval breeding occurring in used tyres, water storage and discarded containers. Most of the insecticides tested remain effective to control Aedes vectors in Sao Tome, except DDT and bendiocarb. These data underline the importance of raising community awareness and implementing routine dengue vector control strategies to prevent further outbreaks in Sao Tome and Principe, and elsewhere in the subregion.

Spatial distribution and insecticide resistance profile of Aedes aegypti and Aedes albopictus in Douala, the most important city of Cameroon.

Prevention and control of Aedes-borne viral diseases such as dengue rely on vector control, including the use of insecticides and reduction of larval sources. However, this is threatened by the emergence of insecticide resistance. This study aimed to update the spatial distribution, the insecticide resistance profile of A. aegypti and A. albopictus and the potential resistant mechanisms implicated in the city of Douala. Immature stages of Aedes were collected in August 2020 in eight neighbourhoods in Douala and reared to adult stages. Adult bioassays, and piperonyl butoxide (PBO) synergist assays were carried out according to World Health Organization recommendations. Expression of some candidate metabolic genes including Cyp9M6F88/87, Cyp9J28a, Cyp9J10 and Cyp9J32 in A. aegypti, and Cyp6P12 in A. albopictus were assessed using qPCR. A. aegypti adults G0 were screened using real time melting curve qPCR analyses to genotype the F1534C, V1016I and V410L Aedes kdr mutations. Overall, A. aegypti is the predominant Aedes species, but analyses revealed that both A. albopictus and A. aegypti coexist in all the prospected neighbourhoods of Douala. High level of resistance was observed to three pyrethroids tested in both Aedes species. In A. aegypti a lower mortality rate was reported to permethrin (5.83%) and a higher mortality rate to deltamethrin (63.74%). Meanwhile, for A. albopictus, lower (6.72%) and higher (84.11%) mortality rates were reported to deltamethrin. Similar analysis with bendiocarb, revealed for A. aegypti a loss of susceptibility. However, in A. albopictus samples, analyses revealed a susceptibility in Logbessou, and confirmed resistance in Kotto (59.78%). A partial recovery of mortality was found to insecticides after pre-exposure to PBO. Cyp6P12 was found significantly overexpressed in A. albopictus permethrin resistant and Cyp9M6F88/87 for A. aegypti deltamethrin resistant. F1534C, V1016I and V410L mutations were detected in A. aegypti from different neighbourhoods and by considering the combination of these three kdr 14 genotypes were found. These findings provide relevant information which should be capitalised in the implementation of arbovirus vector control strategies and insecticide resistance management.

Spatial distribution of insecticide resistant populations of Aedes aegypti and Ae. albopictus and first detection of V410L mutation in Ae. aegypti from Cameroon.

BACKGROUND: Dengue (DENV), chikungunya (CHIKV) and Zika virus (ZIKV), are mosquito-borne viruses of medical importance in most tropical and subtropical regions. Vector control, primarily through insecticides, remains the primary method to prevent their transmission. Here, we evaluated insecticide resistance profiles and identified important underlying resistance mechanisms in populations of Aedes aegypti and Ae. albopictus from six different regions in Cameroon to pesticides commonly used during military and civilian public health vector control operations. METHODS: Aedes mosquitoes were sampled as larvae or pupae between August 2020 and July 2021 in six locations across Cameroon and reared until the next generation, G1. Ae. aegypti and Ae. albopictus adults from G1 were tested following World Health Organization (WHO) recommendations and Ae. aegypti G0 adults screened with real time melting curve qPCR analyses to genotype the F1534C, V1016I and V410L Aedes kdr mutations. Piperonyl butoxide (PBO) assays and real time qPCR were carried out from some cytochrome p450 genes known to be involved in metabolic resistance. Statistical analyses were performed using Chi-square test and generalized linear models. RESULTS: Loss of susceptibility was observed to all insecticides tested. Mortality rates from tests with 0.25% permethrin varied from 24.27 to 85.89% in Ae. aegypti and from 17.35% to 68.08% in Ae. albopictus. Mortality rates for 0.03% deltamethrin were between 23.30% and 88.20% in Ae. aegypti and between 69.47 and 84.11% in Ae. albopictus. We found a moderate level of resistance against bendiocarb, with mortality rates ranging from 69.31% to 90.26% in Ae. aegypti and from 86.75 to 98.95% in Ae. albopictus. With PBO pre-exposure, we found partial or fully restored susceptibility to pyrethroids and bendiocarb. The genes Cyp9M6F88/87 and Cyp9J10 were overexpressed in Ae. aegypti populations from Douala sites resistant to permethrin and deltamethrin. Cyp6P12 was highly expressed in alphacypermethrin and permethrin resistant Ae. albopictus samples. F1534C and V1016I mutations were detected in A. aegypti mosquitoes and for the first time V410L was reported in Cameroon. CONCLUSIONS: This study revealed that Ae. aegypti and Ae. albopictus are resistant to multiple insecticide classes with multiple resistance mechanisms implicated. These findings could guide insecticide use to control arbovirus vectors in Cameroon.