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The most widespread arboviral diseases such as Dengue, Chikungunya, and Zika are transmitted mainly by Aedes mosquitoes. Due to the lack of effective therapeutics for most of these diseases, vector control remains the most effective preventative and control measure. This study investigated and compared the species composition, insecticide susceptibility, and resistance mechanisms in Aedes mosquito populations from a forest reserve converted to an eco-park and a peri-domestic sites in urban Accra, Ghana. Immature Aedes were sampled from the study sites, raised to adults, and exposed to deltamethrin, permethrin, DDT, fenitrothion, bendiocarb, permethrin + PBO, and deltamethrin + PBO using WHO tube assays. Melting curve analyses were performed for F1536C, V1016I, and V410L genetic mutations in surviving and dead mosquitoes following exposure to deltamethrin and permethrin. Microplate assay was used to access enzyme activity levels in adult mosquitoes from both populations. Aedes aegypti was found to be the dominant species from both study populations. The susceptibility test results revealed a high frequency of resistance to all the insecticides except fenitrothion. F1534C mutations were observed in 100% and 97% of mosquitoes from the peri-domestic and forest population, respectively but were associated with pyrethroid resistance only in the forest population (P < 0.0001). For the first time in Aedes mosquitoes in Ghana, we report the existence V410L mutations, mostly under selection only in the forest population (HWE P < 0.0001) and conclude that Aedes vectors in urban Accra have developed resistance to many commonly used insecticides. This information is important for the formulation of vector control strategies for Aedes control in Ghana.

BackgroundDengue, Zika and Chikungunya are Aedes-borne viral diseases that have risen to become great global health concerns in the past years. Several countries in Africa have reported outbreaks of these diseases and despite Ghana sharing borders with some of such countries, it remains free of these outbreaks.Recent studies in Ghana have revealed that there are antibodies and viral RNA of the Dengue virus serotype-2 among individuals in some localities in the Greater Accra Region. This is an indication of a possible silent transmission ongoing in the population, hence the need to assess the risk of transmission of these viruses within the country. This cross-sectional study, therefore, assessed the risk of transmission of Dengue, Zika and Chikungunya viruses in a domestic/peri-domestic (Madina) and a forest (Achimota Forest) population in the Greater Accra Region, Ghana.Methodology/FindingsAll stages of the Aedes mosquito (egg, larvae, pupae and adults) were collected around homes and in the forest area for estimation of risk indices. All eggs and immature stages were reared to adults and morphologically identified. The predominant species of Aedes mosquitoes identified from both sites were Aedes aegypti (98 % in Madina and 98.1% in Achimota forest). Aedes albopictus, an important arbovirus vector, was identified only in Madina at a prevalence of 1.5% but Achimota forest had the higher species diversity. Both study sites recorded high risk indices; Madina: Positive Ovitrap Index = 26.6%, Container Index = 36.8%, House Index = 19.8%, Breteau Index = 70.4%; Achimota: Positive Ovitrap Index = 34.2% and Container Index = 67.9%. RT-PCR to detect the presence of Dengue, Chikungunya and Zika viruses was negative for all pools tested.ConclusionAll entomological risk indicators estimated showed that both sites had a high potential of an outbreak of arboviral diseases following the introduction of these viruses.Author SummaryThe detection of antibodies and viral RNA of the dengue virus serotype 2 in some communities in the urban city of Accra, suggested the possibility of silent transmission of arboviral disease within the city. We assessed the risk of arboviral disease transmission using entomological risk indices. The study was a cross-sectional study conducted in a forest and peri domestic setting located in the southern urban city of Accra.The different stages of the Aedes mosquito were collected and, houses and containers positive for Aedes mosquitoes were also noted. The Breteau (BI), House (HI), Container (CI) and Positive ovitrap (POI) indices were determined. Real Time-PCR was conducted to determine the presence of Dengue, Zika and Chikungunya viruses in the larvae and adults collected.Aedes aegypti was the most common species identified from both sites. Aedes albopictus another competent arbovirus vector was identified in the peri-domestic site. Almost all risk indices recorded for both sites were higher than the WHO thresholds allowed for these indices. However, real time-PCR to detect the presence of Dengue, Chikungunya and Zika viruses was negative.The high entomological risk indicators estimated showed that both sites had a great potential of an outbreak following the introduction of these viruses, and a well-structured surveillance for these vectors is highly recommended. The detection of the presence of Ae. albopictus, an invasive species is also of great concern.

Some bacteria species found in the mosquito midgut have demonstrated their role in interrupting the development of Plasmodium within the midgut of the Anopheles mosquito and have been identified as potential candidates for novel bacteria-mediated disease control. However, to use these bacteria successfully in biocontrol mechanisms their effect on the fitness of the vector into which they have been introduced has to be evaluated. This study investigated the effect of two such bacteria candidates, Enterobacter cloacae and Serratia marcescens, on Anopheles gambiae s.l. fitness. Pupae and larvae of Anopheles gambiae s.l. mosquitoes were collected by dipping method and reared to adults. The effect of these bacteria on mosquito fitness was assessed by reintroducing isolates of each bacteria separately into antibiotic-treated female adult mosquitoes through sugar meal. Wild type (non-antibiotic-treated) mosquitoes and those antibiotic-treated with no bacteria reintroduction were used as controls. The mosquitoes were monitored on longevity/survival, fecundity, hatch rate, and larval survival. The antibiotic-treated adult mosquitoes had reduced life span with median survival of 14 days while the bacteria-reintroduced groups and the wild type survived to day 22 (p< 0.0001). Treatment with Enterobacter and Serratia did not affect the average egg deposition (p>0.05) but they affected hatch rates positively (p = 0.008). There was, however, some evidence that suggests Enterobacter could have a positive effect on larval development (p < 0.0001). With no observed negative effect on survival/longevity of Anopheles gambiae, introducing E. cloacae and S. marcescens in future bacteria-associated control strategies is unlikely to result in mosquitoes that will be outlived by the wild population. This, however, requires evaluations under field conditions.

Novel ideas for control of mosquito-borne disease include the use of bacterial symbionts to reduce transmission. Bacteria belonging to the family Enterobacteriaceae isolated from mosquito midgut have shown promise in limiting Plasmodium intensity in the Anopheles vector. However, the mechanism of interaction between bacteria and parasite remains unclear. This study aimed at screening bio-products of two bacteria candidates for their anti-Plasmodial effects on mosquito stages of P. falciparum. Enterobacter cloacae and Serratia marcescens were isolated from field-caught Anopheles gambiae s.l. Spent media from liquid cultures of these bacteria were filtered, lyophilized and dissolved in sterile phosphate buffered saline (PBS). The re-dissolved bacterial products were added to gametocytaemic blood meals and fed to An. gambiae mosquitoes via membrane feeders. Control groups were fed on infected blood with or without lyophilized LB medium. The effect of the products on the infection prevalence and intensity of P. falciparum in mosquitoes was assessed by dissecting mosquito midguts and counting oocysts 10-11 days post-infection. S. marcescens bio-products elicited significant reduction in the number of mosquitoes infected (P=4.02 x10-5) with P. falciparum and the oocyst intensity (P-16) than E. cloacae products (P>0.05 for both prevalence and intensity) compared to the control (lyophilized LB medium). These data support the use of bioproducts released by S. marcescens for malaria control based on transmission blocking in the vector.