Genetic Polymorphism and Phylogenetics of Aedes aegypti from Sudan Based on ND4 Mitochondrial Gene Variations.

This study investigated the genetic differences between Aedes aegypti subspecies (Aedes aegypti aegypti (Aaa) and Aedes aegypti formosus (Aaf)) from Sudan using the NADH dehydrogenase subunit 4 (ND4) mitochondrial gene marker. Nineteen distinct haplotypes of the ND4 were identified in female Aedes aegypti mosquitoes from the study sites. The phylogenetic relationship of the 19 ND4 haplotypes was demonstrated in a median-joining haplotype network tree with Aaa and Aaf populations found to share three haplotypes. The genetic variance (Pairwise FST values) was estimated and found to range from 0.000 to 0.811. Isolation by distance test revealed that geographical distance was correlated to genetic variation (coefficient value (r) = 0.43). The Polar maximum likelihood tree showed the phylogenetic relationship of 91 female Aaa and Aaf from the study sites, with most of the Aaf haplotypes clustered in one group while most of the Aaa haplotypes gathered in another group, but there was an admixture of the subspecies in both clusters, especially the Aaa cluster. The Spatial Analysis of Molecular Variance (SAMOVA) test revealed that the eight populations clustered into two phylogeographic groups/clusters of the two subspecies populations. The 2 Aedes aegypti subspecies seemed not to be totally separated geographically with gene flow among the populations.

Distribution and Genetic Diversity of Aedes aegypti Subspecies across the Sahelian Belt in Sudan.

Aedes aegypti is the most important arboviral disease vector worldwide. In Africa, it exists as two morphologically distinct forms, often referred to as subspecies, Aaa and Aaf. There is a dearth of information on the distribution and genetic diversity of these two forms in Sudan and other African Sahelian region countries. This study aimed to explore the distribution and genetic diversity of Aedes aegypti subspecies using morphology and Cytochrome oxidase-1 mitochondrial marker in a large Sahelian zone in Sudan. An extensive cross-sectional survey of Aedes aegypti in Sudan was performed. Samples collected from eight locations were morphologically identified, subjected to DNA extraction, amplification, sequencing, and analyses. We classified four populations as Aaa and the other four as Aaf. Out of 140 sequence samples, forty-six distinct haplotypes were characterized. The haplotype and nucleotide diversity of the collected samples were 0.377-0.947 and 0.002-0.01, respectively. Isolation by distance was significantly evident (r = 0.586, p = 0.005). The SAMOVA test indicated that all Aaf populations are structured in one group, while the Aaa clustered into two groups. AMOVA showed 53.53% genetic differences within populations and 39.22% among groups. Phylogenetic relationships indicated two clusters in which the two subspecies were structured. Thus, the haplotype network consisted of three clusters.

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.

Exposure of a diurnal mosquito vector to floral mimics: Foraging responses, feeding patterns, and significance for sugar bait technology.

Food location by mosquitoes is mediated by resource-derived olfactory and visual signals. Smell sensation is intermittent and dependent on the environment, whereas visual signals are continual and precede olfactory cues. Success of mosquito bait technology, where olfactory cues are used for attraction, is being impeded by reduced attractiveness. Despite proof that mosquitoes respond to colored objects, including those mimicking floral shape, and that they can discriminate among flowers, the impacts of artificial flowers on foraging remain unexplored. Using artificial flowers with sugar rewards, we examined the foraging responses of Aedes aegypti to various colors in equal choice bioassays. Starved adults were exposed to single flowers with petals of a given color (Single Blue Flowers [SBFs]; Single Red Flowers [SRFs]; Single Yellow Flowers [SYFs]; Single Pink Flowers [SPIFs]; and Single Purple Flowers [SPFs]) and two others with white petals (SWFs). Discrepancies in response time, visitation, feeding, and resting of both sexes were compared between colored flowers and SWFs. Ae. aegypti exhibited shorter response times to colored flowers compared to SWFs, but this behavior was mostly seen for SBFs or SYFs in females, and SRFs, SYFs, SPIFs, or SPFs in males. When provided an option to land on colored flowers and SWFs, female visitation occurred at high rates on SBFs, SRFs, SYFs, SPIFs, and SPFs; for males, this preference for colored flowers was seen to a lesser degree on SBF and SPIFs. Both sexes exhibited preference for colored flowers as sugar sources, but with different patterns: SPIFs, SRFs, SYFs, and SPFs for females; SYFs, SPFs, SPIFs and SRFs for males. Females preferentially rested on colored flowers when in competition with SWFs, but this preference was more pronounced for SPFs, SRFs, and SBFs. Males exhibited an increased preference for SRFs, SPFs, and SYFs as resting sites. Our results indicated the attraction of Ae. aegypti to rewarding artificial flowers, in some cases in ways similar to live flowering plants. The discovery that both male and female Ae. aegypti can feed on nectar mimics held by artificial flowers opens new avenues for improving sugar bait technology and for developing new attract-and-kill devices.

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.

Indirect effects of cigarette butt waste on the dengue vector Aedes aegypti (Diptera: Culicidae).

Despite major insecticide-based vector control programs, dengue continues to be a major threat to public health in urban areas. The reasons for this failure include the emergence of insecticide resistance and the narrowing of the spectrum of efficient products. Cigarette butts (CBs), the most commonly discarded piece of waste, also represent a major health hazard to human and animal life. CBs are impregnated with thousands of chemical compounds, many of which are highly toxic and none of which has history of resistance in mosquitoes. This study was performed to examine whether exposure to CB alters various biological parameters of parents and their progeny. We examined whether the mosquito changes its ovipositional behaviors, egg hatching, reproductive capacity, longevity and fecundity in response to CB exposure at three different concentrations. Females tended to prefer microcosms containing CBs for egg deposition than those with water only. There were equivalent rates of eclosion success among larvae from eggs that matured in CB and water environments. We also observed decreased life span among adults that survived CB exposure. Extracts of CB waste have detrimental effects on the fecundity and longevity of its offspring, while being attractive to its gravid females. These results altogether indicate that CB waste indirectly affect key adult life traits of Aedes aegypti and could conceivably be developed as a novel dengue vector control strategy, referring to previously documented direct toxicity on the larval stage. But this will require further research on CB waste effects on non-target organisms including humans.