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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.

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.

BackgroundAedesborne viral diseases, notably dengue, are increasingly reported in Cameroon withAedes aegyptibeing a major vector. Data on insecticide resistance of this vector and underlying mechanisms needed for outbreak preparedness remain scarce in Cameroon. Here, we present the nationwide distribution of insecticide resistance inAe. aegyptiand investigate the potential resistance mechanisms involved.MethodsImmature stages ofAe. aegyptiwere collected between March and July 2017 in 13 locations across Cameroon and reared until G1/G2/G3 generation. Larval, adult bioassays, and piperonyl butoxide (PBO) synergist assays were carried out according to World Health Organization guidelines. F1534C mutation was genotyped using allele specific polymerase chain reaction in field collected adults (Go) and the polymorphism of the sodium channel gene was assessed. Theχ2test was used to compare the mortality rate between bioassays with insecticides only and bioassays after preexposure to PBO synergist.ResultsLarval bioassay revealed that all the three populations tested with temephos were susceptible. Adult bioassays showed a good level of susceptibility toward both pyrethroids tested, 0.25% permethrin and 0.05% deltamethrin, with six out of 10 populations susceptible. However, two populations (Douala and Edéa) were resistant (deltamethrin [73.2–92.5% mortality], permethrin [2.6–76.3% mortality]). The resistance to 4% dichlorodiphenyltrichloroethane was observed in four out of 10 populations tested (16.8–87.1% mortality). Resistance was also reported to carbamates including 0.1% propoxur (60.8–87.1% mortality) and to 0.1% bendiocarb (82.9% mortality). All populations tested were fully susceptible to 1% fenitrothion. A partial recovery of susceptibility was observed in the pyrethroid resistant population of Douala after pre-exposed to PBO suggesting the implication of cytochrome P450 monoxygenases permethrin resistance. Genotyping and sequencing detected the F1534Ckdrmutation in the two pyrethroid resistant locations of Edéa and Douala, with allelic frequency of 3.3% and 33.3% respectively. However, the high genetic diversity of the sodium channel gene supports the recent introduction of this mutation in Cameroon.ConclusionsThis study revealed the contrasting resistance profiles to insecticides ofAe. aegyptipopulations in Cameroon suggesting that, instead of a unique nationwide control approach, a regionally adapted strategy will be needed to control this vector. The localised distribution of the F1534Ckdrmutation supports this region-specific control strategy.

Background In the Republic of Congo, with two massive outbreaks of chikungunya observed this decade, little is known about the insecticide resistance profile of the two major arbovirus vectors Aedes aegypti and Aedes albopictus. Here, we established the resistance profile of both species to insecticides and explored the resistance mechanisms to help Congo to better prepare for future outbreaks. Methods Immature stages of Ae. aegypti and Ae. albopictus were sampled in May 2017 in eight cities of the Republic of the Congo and reared to adult stage. Larval and adult bioassays, and synergist (piperonyl butoxide [PBO]) assays were carried out according to WHO guidelines. F1534C mutation was genotyped in field collected adults in both species and the polymorphism of the sodium channel gene assessed in Ae. aegypti. Results All tested populations were susceptible to temephos after larval bioassays. A high resistance level was observed to 4% DDT in both species countrywide (21.9–88.3% mortality). All but one population (Ae. aegypti from Ngo) exhibited resistance to type I pyrethroid, permethrin, but showed a full susceptibility to type II pyrethroid (deltamethrin) in almost all locations. Resistance was also reported to 1% propoxur in Ae. aegypti likewise in two Ae. albopictus populations (Owando and Ouesso), and the remaining were fully susceptible. All populations of both species were fully susceptible to 1% fenitrothion. A full recovery of susceptibility was observed in Ae. aegypti and Ae. albopictus when pre-exposed to PBO and then to propoxur and permethrin respectively. The F1534C kdr mutation was not detected in either species. The high genetic variability of the portion of sodium channel spanning the F1534C in Ae. aegypti further supported that knockdown resistance probably play no role in the permethrin resistance. Conclusions Our study showed that both Aedes species were susceptible to organophosphates (temephos and fenitrothion), while for other insecticide classes tested the profile of resistance vary according to the population origin. These findings could help to implement better and efficient strategies to control these species in the Congo in the advent of future arbovirus outbreaks.

Background: The arbovirus vector, Aedes albopictus, originating from Asia, has recently invaded African countries, including the Republic of the Congo, where it was associated with a chikungunya outbreak. Up until now, little was known about its distribution in relation to the native Aedes aegypti and how the invasion will modify the epidemiology of arboviral diseases. Here, we assessed the current distribution of Ae. albopictus and Ae. aegypti in the Republic of the Congo and explored the genetic diversity of the invading species, Ae. albopictus. Methods: Immature stages of Aedes were collected in nine locations in the Republic of the Congo in 2017 following a north-south transect and reared to adult stage. Adults were morphologically identified, counted and grouped according to species and location. Genetic diversity of Ae. albopictus was assessed by analyzing the cytochrome oxidase I ( COI) gene. Results: Ae. albopictus and Ae. aegypti were found together across the country in all the locations investigated. The invasive species is predominant over the native species in all locations except Brazzaville, suggesting that Ae. albopictus is displacing Ae. aegypti across Congo. When comparing the species distributions across the two largest cities, Brazzaville and Pointe Noire, Ae. albopictus was more prevalent than Ae. aegypti in the suburbs whereas the opposite situation was reported in the city centre. Mitochondrial DNA analysis revealed very low genetic diversity of Ae. albopictus with only three haplotypes recorded across the country supporting the recent introduction of this species in the Republic of the Congo. Phylogenetic tree analysis revealed that Ae. albopictus from Congo originated from other tropical Asian countries such as China, likely as a result of increasing trade links. Conclusion: These findings are important for the implementation of vector control strategies and can serve as a foundation for further research on these vectors in the country.

Background:The arbovirus vector,Aedes albopictus,originating from Asia, has recently invaded African countries, including the Republic of the Congo, where it was associated with a chikungunya outbreak. Up until now, little was known about its distribution in relation to the nativeAedes aegyptiand how the invasion will modify the epidemiology of arboviral diseases. Here, we assessed the current distribution ofAe. albopictusandAe. aegyptiin the Republic of the Congo and explored the genetic diversity of the invading species,Ae. albopictus.Methods:Immature stages ofAedeswere collected in nine locations in the Republic of the Congo in 2017 following a north-south transect and reared to adult stage. Adults were morphologically identified, counted and grouped according to species and location. Genetic diversity ofAe. albopictuswas assessed by analyzing the cytochrome oxidase I (COI) gene.Results:Ae.albopictusandAe. aegyptiwere found together across the country in all the locations investigated. The invasive species is predominant over the native species in all locations except Brazzaville, suggesting thatAe. albopictusis displacingAe. aegyptiacross Congo. When comparing the species distributions across the two largest cities, Brazzaville and Pointe Noire,Ae. albopictuswas more prevalent thanAe. aegyptiin the suburbs whereas the opposite situation was reported in the city centre. Mitochondrial DNA analysis revealed very low genetic diversity ofAe. albopictuswith only three haplotypes recorded across the country supporting the recent introduction of this species in the Republic of the Congo. Phylogenetic tree analysis revealed thatAe. albopictusfrom Congo originated from other tropical Asian countries such as China, likely as a result of increasing trade links.Conclusion:These findings are important for the implementation of vector control strategies and can serve as a foundation for further research on these vectors in the country.