Temporal and spatial trends in insecticide resistance in Anopheles arabiensis in Sudan: outcomes from an evaluation of implications of insecticide resistance for malaria vector control

BACKGROUND: Long-lasting insecticidal nets (LLINs) (with pyrethroids) and indoor residual spraying (IRS) are the cornerstones of the Sudanese malaria control program. Insecticide resistance to the principal insecticides in LLINs and IRS is a major concern. This study was designed to monitor insecticide resistance in Anopheles arabiensis from 140 clusters in four malaria-endemic areas of Sudan from 2011 to 2014. All clusters received LLINs, while half (n = 70), distributed across the four regions, had additional IRS campaigns. METHODS: Anopheles gambiae (s.l.) mosquitoes were identified to species level using PCR techniques. Standard WHO insecticide susceptibility bioassays were carried out to detect resistance to deltamethrin (0.05%), DDT (4%) and bendiocarb (0.1%). TaqMan assays were performed on random samples of deltamethrin-resistant phenotyped and pyrethrum spray collected individuals to determine Vgsc-1014 knockdown resistance mutations. RESULTS: Anopheles arabiensis accounted for 99.9% of any anopheline species collected across all sites. Bioassay screening indicated that mosquitoes remained susceptible to bendiocarb but were resistance to deltamethrin and DDT in all areas. There were significant increases in deltamethrin resistance over the four years, with overall mean percent mortality to deltamethrin declining from 81.0% (95% CI: 77.6-84.3%) in 2011 to 47.7% (95% CI: 43.5-51.8%) in 2014. The rate of increase in phenotypic deltamethrin-resistance was significantly slower in the LLIN + IRS arm than in the LLIN-only arm (Odds ratio 1.34; 95% CI: 1.02-1.77). The frequency of Vgsc-1014F mutation varied spatiotemporally with highest frequencies in Galabat (range 0.375-0.616) and New Halfa (range 0.241-0.447). Deltamethrin phenotypic-resistance correlated with Vgsc-1014F frequency. CONCLUSION: Combining LLIN and IRS, with different classes of insecticide, may delay pyrethroid resistance development, but the speed at which resistance develops may be area-specific. Continued monitoring is vital to ensure optimal management and control

Coffee and its waste repel gravid Aedes albopictus females and inhibit the development of their embryos

Background Dengue is a prevalent arboviral disease and the development of insecticide resistance among its vectors impedes endeavors to control it. Coffee is drunk by millions of people daily worldwide, which is associated with the discarding of large amounts of waste. Coffee and its waste contain large amounts of chemicals many of which are highly toxic and none of which have a history of resistance in mosquitoes. Once in solution, coffee is brownish in colour, resembling leaf infusion, which is highly attractive to gravid mosquitoes. To anticipate the environmental issues related to the increasing popularity of coffee as a drink, and also to combat insecticide resistance, we explored the deterrence potentials of coffee leachates against the ovipositing and embryonic stages of the dengue vector, Aedes albopictus. Methods In a series of choice, no-choice, and embryo toxicity bioassays, we examined changes in the ovipositional behaviours and larval eclosion of Ae. albopictus in response to coffee extracts at different concentrations. Results Oviposition responses were extremely low when ovicups holding highly concentrated extract (HCE) of coffee were the only oviposition sites. Gravid females retained increased numbers of mature eggs until 5 days post-blood feeding. When provided an opportunity to oviposit in cups containing coffee extracts and with water, egg deposition occurred at lower rates in those containing coffee, and HCE cups were far less attractive to females than those containing water only. Females that successfully developed in a coffee environment preferentially oviposited in such cups when in competition with preferred oviposition sites (water cups), but this trait did not continue into the fourth generation. Larval eclosion occurred at lower rates among eggs that matured in a coffee environment, especially among those that were maintained on HCE-moistened substrates. Conclusions The observations of the present study indicate a pronounced vulnerability of Ae. albopictus to the presence of coffee in its habitats during the early phases of its life cycle. The observations that coffee repels gravid females and inhibits larval eclosion provide novel possibilities in the search for novel oviposition deterrents and anti-larval eclosion agents against dengue vectors.This work was supported by funds (No. 096010) from the Central Research Institute of Fukuoka University and “Long Term Research Grant (LRGS) for Infectious Diseases, 2011 – 2014, Ministry of Higher Education, Malaysia” and USM (304/Pbiology/650575/U112)

Contribution of public places in proliferation of dengue vectors in Penang Island, Malaysia

Objective: To determine abundance, distribution and diversity of potential breeding container habitats of the dengue vectors in public places including schools, restaurants, mosques and parks in southwest areas of Penang Island, Malaysia. Methods: Premises at restaurants, schools, parks and mosques were surveyed simultaneously and inspected visually for container habitats and production of immature mosquitoes from March 2015 to March 2016. Abundance (mean ± SE) of breeding containers between sites was compared using One-way ANOVA. Independent sample t-test was used to compare total number of Aedes albopictus (Ae. albopictus) and Aedes aegypti (Ae. aegypti) surveyed. Results: The surveyed locations yielded a total of 3741 breeding containers and 19537 immature mosquitoes from four areas. Concurrent artificial and natural containers produced 78.4% immature Ae. albopictus and 6.3% Ae. aegypti mosquitoes in wet season, with 14.2% Ae. albopictus and 1.1% Ae. aegypti mosquitoes in dry season. Artificial containers accounted for 98.1% of the total containers recorded, with restaurants being the most productive locations (8012) and schools being the least productive (2234). Conclusions: It was concluded that public places are good sources of potential container habitats of Aedes mosquitoes in Penang Island, Malaysia and Ae. albopictus has exclusively replaced the home-grown Ae. aegypti even in urban areas. Therefore, treatment of artificial containers in such locations is critical in Aedes mosquito control campaigns during dengue outbreaks

Contribution of public places in proliferation of dengue vectors in Penang Island, Malaysia

Objective: To determine abundance, distribution and diversity of potential breeding container habitats of the dengue vectors in public places including schools, restaurants, mosques and parks in southwest areas of Penang Island, Malaysia. Methods: Premises at restaurants, schools, parks and mosques were surveyed simultaneously and inspected visually for container habitats and production of immature mosquitoes from March 2015 to March 2016. Abundance (mean ± SE) of breeding containers between sites was compared using One-way ANOVA. Independent sample t-test was used to compare total number of Aedes albopictus (Ae. albopictus) and Aedes aegypti (Ae. aegypti) surveyed. Results: The surveyed locations yielded a total of 3. 741 breeding containers and 19. 537 immature mosquitoes from four areas. Concurrent artificial and natural containers produced 78.4% immature Ae. albopictus and 6.3% Ae. aegypti mosquitoes in wet season, with 14.2% Ae. albopictus and 1.1% Ae. aegypti mosquitoes in dry season. Artificial containers accounted for 98.1% of the total containers recorded, with restaurants being the most productive locations (8. 012) and schools being the least productive (2. 234). Conclusions: It was concluded that public places are good sources of potential container habitats of Aedes mosquitoes in Penang Island, Malaysia and Ae. albopictus has exclusively replaced the home-grown Ae. aegypti even in urban areas. Therefore, treatment of artificial containers in such locations is critical in Aedes mosquito control campaigns during dengue outbreaks

Sex before or after blood feeding : Mating activities of Aedes aegypti males under conditions of different densities and female blood feeding opportunities

Blood feeding and mating are critical behaviors that regulate both mosquito population maintenance and disease transmission. However, our understanding of mosquito mating systems remains incomplete. One of the most critical issues is a lack of understanding regarding how and where males and females encounter one another. This study was performed to investigate changes in key mating behaviors of Ae. aegypti relative to female blood feeding opportunities, taking into account male density. We compared courtship latency and copulation activity between single and pooled males in a range of assays performed in the presence or absence of a blood source and after blood feeding. The time taken by grouped males to initiate courtship in the presence of a host was much shorter than that in single males. There was no significant difference in courtship latency between pooled and single males in the absence of a blood source or after blood feeding. At low male density, the presence of the host and blood meal ingestion provided better conditions for copulation. At high male density, however, copulation activity was decreased after blood feeding, but remained high regardless of the presence or absence of the host. In addition to providing insight into the mating ecology of Aedes aegypti, this study indicated that the presence of a blood source influences how males encounter and copulate with females. The observation that copulation activity decreases after blood feeding when males are numerous provides new avenues for improving mass release programs of sterile mosquitoes

Biting rhythm and demographic attributes of Aedes albopictus (Skuse) females from different urbanized settings in Penang Island, Malaysia under uncontrolled laboratory conditions.

Urbanization could potentially modify Aedes albopictus' ecology by changing the dynamics of the species, and affecting their breeding sites due to environmental changes, and thus contribute to dengue outbreaks. Thus, this study was conducted to evaluate the biting rhythm, fecundity and longevity of adult female Ae. albopictus in relation to urbanization strata; urban, suburban and rural areas in Penang Island, Malaysia. The experiments were done in comparison to a laboratory strain. Twenty-four hours biting activity of all the mosquito strains showed a clear bimodal biting activity, with morning and evening twilight peaks. The interaction effect between biting time and mosquito strains was not significant. Meanwhile, differences in fecundity among mosquito strains were statistically significant (F(3,442) = 10.559, P < 0.05) with urban areas having higher mean number of eggs (mean = 107.69, standard error = 3.98) than suburban (mean = 94.48, standard error = 5.18), and rural areas (mean = 72.52, standard error = 3.87). Longevity of adult females were significantly higher (F(3,441) = 31.259, P < 0.05) for mosquito strains from urban areas compared to the other strains. These findings would provide crucial information for the planning of control programs in Malaysia, particularly Penang

Population genetic structure of Aedes aegypti subspecies in selected geographical locations in Sudan

Abstract Although knowledge of the composition and genetic diversity of disease vectors is important for their management, this is limiting in many instances. In this study, the population structure and phylogenetic relationship of the two Aedes aegypti subspecies namely Aedes aegypti aegypti (Aaa) and Aedes aegypti formosus (Aaf) in eight geographical areas in Sudan were analyzed using seven microsatellite markers. Hardy–Weinberg Equilibrium (HWE) for the two subspecies revealed that Aaa deviated from HWE among the seven microsatellite loci, while Aaf exhibited departure in five loci and no departure in two loci (A10 and M201). The Factorial Correspondence Analysis (FCA) plots revealed that the Aaa populations from Port Sudan, Tokar, and Kassala clustered together (which is consistent with the unrooted phylogenetic tree), Aaf from Fasher and Nyala populations clustered together, and Gezira, Kadugli, and Junaynah populations also clustered together. The Bayesian cluster analysis structured the populations into two groups suggesting two genetically distinct groups (subspecies). Isolation by distance test revealed a moderate to strong significant correlation between geographical distance and genetic variations (p = 0.003, r = 0.391). The migration network created using divMigrate demonstrated that migration and gene exchange between subspecies populations appear to occur based on their geographical proximity. The genetic structure of the Ae. aegypti subspecies population and the gene flow among them, which may be interpreted as the mosquito vector's capacity for dispersal, were revealed in this study. These findings will help in the improvement of dengue epidemiology research including information on the identity of the target vector/subspecies and the arboviruses vector surveillance program