Co-occurrence of multiple kdr mutations (F1534C, V1016I, V410L) in Aedes aegypti from coastal areas in Ghana and assessment of the role of mosquito coil in causing pyrethroid resistance.

The rapid spread of knockdown-resistance (kdr) mutations in Africa calls for monitoring and investigation into the cause of pyrethroid resistance to inform management strategies. This study investigated the pyrethroid resistance profile of Aedes aegypti from coastal towns in Ghana and the impact of mosquito coil, a popular household pyrethroid-based anti-mosquito tool, on the development of pyrethroid resistance. Susceptibility to deltamethrin and the presence of kdr mutations was determined in adult female mosquitoes reared from larvae. Furthermore, the LT50 of a mosquito coil (0.08% meperfluthrin) against a laboratory colony was determined, and the value was used as a sublethal dose in an experimental study. The laboratory colony of Ae. aegypti was exposed to the sublethal dose of the coil once per generation for six generations (F6). The susceptibility of the exposed colony to deltamethrin (0.05%) was determined. The Ae. aegypti populations from the coastal towns were resistant to deltamethrin with co-occurrence of F1534C, V1016I and V410L kdr mutations. In the experimental study, the LT50 (95% CI) of the selected colony against the coil rose from 8 minutes (95% CI; 6-9) at F0 to 28 minutes (95% CI; 23-34) at F6. Nonetheless, deltamethrin caused similar mortalities in the selected and control colonies. The mutant allele frequencies of 1534C and 410L were similar but 1016I was higher in the selected colony (17%) than in the control (5%). However, the increased tolerance to the coil and high mutant allele frequency of 1016I in the selected colony did not affect the mosquito's resistance level to deltamethrin insecticide. Further study is needed to elucidate the role of pyrethroid-based mosquito coils in the development of insecticide resistance in mosquito vectors.

Water quality assessment in mosquito breeding habitats based on dissolved organic matter and chlorophyll measurements by laser-induced fluorescence spectroscopy.

Rapid urbanization and its associated pollution can affect water quality in mosquito breeding habitats and, as a result, the ecology and control of mosquito vectors. To understand the effects of pollution on mosquito vectors, an accurate assessment of water quality in breeding habitats is needed. Presently, water quality assessment of mosquito breeding habitats is usually based on the measurement of individual physicochemical parameters. However, several parameters are sometimes difficult to interpret or may not give a clear picture of the overall water quality of the breeding habitats, especially when the pollutants are in complex mixtures. This study employed the use of Laser-Induced Fluorescence (LIF) spectroscopy to assess water quality in breeding habitats of Anopheles, Aedes, and Culex mosquitoes in urban areas in Cape Coast, Ghana. The LIF spectra, using a 445-nm diode laser, were measured from field-collected water samples in the laboratory. The LIF spectra showed the presence of dissolved organic matter (DOM) and chlorophyll in the breeding habitats. The DOM and chlorophyll fluorescence signals were normalised by the Raman vibrational signals to determine water quality in each habitat. The overall water quality was better in Aedes breeding habitats than in Anopheles and Culex breeding habitats. The poor water quality in Anopheles and Culex breeding habitats was due to the presence of high fulvic acid and chlorophyll content, which often reflect pollutants from anthropogenic sources. Anopheles and Aedes habitats were made up of mainly An. coluzzii and Ae. aegypti respectively while Culex species were identified to genus level. The results add up to the growing concern about the breeding of Anopheles in polluted habitats. The study demonstrated for the first time the ability of LIF spectroscopy to assess water quality in mosquito breeding habitats.

Preliminary assessment of the potential role of urbanization in the distribution of carbamate and organophosphate resistant populations of Culex species in Ghana.

BACKGROUND: Besides its role as a pathogen vector, Culex species also indirectly promotes the transmission of malaria if the use of bed nets or indoor residual spraying is discontinued due to a lack of insecticide efficacy against it. A recent survey revealed widespread occurrence of pyrethroid resistance among urban populations of this mosquito in Ghana. In order to plan and implement insecticide-based resistance management strategies, this study was carried out to assess resistance status of Culex species to organophosphate and carbamate in urban areas in Ghana and the possible mechanisms involved as well as environmental factors associated with its distribution. METHODS: Mosquito larvae were sampled from various land use and ecological settings and in different seasons. In adults, susceptibility to organophosphates (fenitrothion, malathion) and carbamates (propoxur, bendiocarb) were determined. Mixed function oxidase (MFO) and α- and ß-esterase assays, as well as a PCR diagnostic assay to determine ace1 mutation were performed in individual mosquitoes. RESULTS: Culex quinquefasciatus as well as C. decens and other unidentified Culex species were found breeding in polluted water bodies in the study sites. Across all sites and seasons, carbamate induced mortality was 94.1% ± 15.4 whereas mortality caused by organophosphate was 99.5% ± 2.2. In addition, ace1 mutation and high levels of esterases were detected in some of the mosquito populations. There was a strong correlation between susceptibility status of the mosquitoes and the level of absorbance of ß-esterase (Pearson r=- 0.841, p=0.004). CONCLUSIONS: The study found low prevalence of resistance to carbamate and organophosphate insecticides among Culex species from Ghana. However, there were populations with ace1 mutations and high levels of esterases, which can confer high resistance to these classes of insecticides. Thus, it is important to monitor activities or behaviour that has the potential to select for carbamate and organophosphate resistance populations.

Assessment of anti mosquito measures in households and resistance status of Culex species in urban areas in southern Ghana: implications for the sustainability of ITN use.

OBJECTIVE: To determine resistance status of Culex species to different class of insecticides and assess the major anti-mosquito strategies employed by urban households and their possible effects on malaria acquisition. METHODS: Structured questionnaires were randomly administered to obtain information on demographic characteristics, measures that people use to prevent mosquito bites and their perception of where mosquitoes breed in their communities. Adult susceptibility assays were also carried out using WHO discriminating dosages of four insecticides from different chemical classes against Culex species. RESULTS: Majority of respondents (54.75%) preferred the use of domestic insecticides in the form of aerosols and coils. Among households that used domestic insecticides, the use of coil was most frequent (62.9%) with a mean (95% CI) of 9.18 (8.99, 9.37) coils per week. Strong level of pyrethroid-resistance and multiple insecticide resistance in Culex species were also detected in some of the study sites. CONCLUSIONS: The excessive use of domestic insecticides and high level of resistance in Culex species observed in the study area has implications for the ITN component of the nation's malaria control program in more subtle ways. People will lose interest in the use of ITN when it fails to protect users from bites of resistant Culex species. Excessive use of domestic insecticides may also select resistance in both malaria vectors and Culex species. On this account we recommend that nuisance mosquitoes must be controlled as part of malaria control programs to improve acceptance and utilization of ITN.

Aqueous neem extract versus neem powder on Culex quinquefasciatus: implications for control in anthropogenic habitats.

Control programs using conventional insecticides to target anthropogenic mosquito habitats are very expensive because these habitats are widespread, particularly in cities of most African countries. Additionally, there are serious environmental concerns regarding large-scale application of most conventional insecticides. Clearly there is a need for alternative methods that are more effective, less expensive, and environmentally friendly. One such method would be the application of preparations made from parts of the neem tree, Azadirachta indica A. Jussieu (Sapindales: Meliaceae). In this study, aqueous crude extracts and crude powder were prepared from different parts of neem, and the efficacies of the preparations on juvenile stages of Culex quinquefasciatus Say (Diptera: Culicidae) were evaluated in the laboratory. When larvae were exposed to a concentration of 0.1 g/mL extract for 24 hours, percent mean mortality (± SE) was 72.7 plusmn; 1.8 for the bark, 68.7 ± 1.6 for fruits and 60 ± 1.6 for leaves. These means were not significantly different (χ(2) = 4.12; df = 2; p = 0.127). At a concentration of 0.01 g/mL, > 95% of the larvae died within 24 hours of exposure to powdered neem leaf, but it took 120 hours to reach the same level of larval mortality in aqueous leaf extract. The crude extract slowly inhibited the growth and development of mosquitoes while the crude powder acted more as a barrier; the mosquitoes probably died from suffocation. However, both types of preparations can be made and used by local people to control mosquito breeding in anthropogenic habitats, especially in urbanized areas.