Dataset Information

First detection of N1575Y mutation in pyrethroid resistant Anopheles gambiae in Southern Cote d'Ivoire.

ABSTRACT: Background. The intensification of insecticide use for both public health and agriculture in Africa has contributed to growing insecticide resistance. Today, resistance to World Health Organization (WHO)-approved insecticide classes is widespread. In an agricultural area of Southern Côte d'Ivoire, the main malaria vector Anopheles coluzzii shows multiple resistance across insecticides mediated by both target site mutation and metabolic mechanisms. To plan new vector control strategies and avert future resistance liabilities caused by cross-resistance mechanisms extant within populations, it is crucial to monitor the development and spread of both resistance and mechanisms. Methods. Larvae of Anopheles gambiae were collected from natural breeding sites in Tiassalé and Elibou, between April and November 2016 and raised to adults . Adult female non-blood fed mosquitoes, three to five days old, were exposed to deltamethrin in WHO bioassays. Extracted DNA samples from exposed mosquitoes were used for species characterisation and genotyping. Results. Most adult An. gambiae tested were resistant to deltamethrin, with mortality rates of only 25% in Tiassalé and 4.4% in Elibou. Molecular analysis of DNA from samples tested showed the presence of both An. coluzzii and An. gambiae s.s in Elibou and only An. coluzzii for Tiassalé. As previously, the L1014F kdr mutation was present at high frequency (79%) in Tiassalé and the L1014S mutation was absent. The N1575Y mutation, which amplifies resistance conferred by L1014F was detected in a single unique individual from a Tiassalé An. coluzzii female whereas in Elibou 1575Y was present in 10 An. gambiae s.s, but not in An. coluzzii. Conclusion. This is the first report of the N1575Y mutation in Côte d'Ivoire, and as in other populations, it is found in both dominant West African malaria vector species. Continued monitoring of N1575Y is underway, as are studies to elucidate its contribution to the resistance of local vector populations.

SUBMITTER: Edi AVC

PROVIDER: S-EPMC5627500 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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First detection of N1575Y mutation in pyrethroid resistant Anopheles gambiae in Southern Côte d'Ivoire.

Edi Ako Victorien Constant AVC N'Dri Bedjou Prisca BP Chouaibou Mouhamadou M Kouadio Fondjo Behi FB Pignatelli Patricia P Raso Giovanna G Weetman David D Bonfoh Bassirou B

Wellcome open research 20170824

Background. The intensification of insecticide use for both public health and agriculture in Africa has contributed to growing insecticide resistance. Today, resistance to World Health Organization (WHO)-approved insecticide classes is widespread. In an agricultural area of Southern Côte d'Ivoire, the main malaria vector Anopheles coluzzii shows multiple resistance across insecticides mediated by both target site mutation and metabolic mechanisms. To plan new vector control strateg ...[more]

PMID: 29018842

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Project description:BackgroundLong-lasting insecticidal nets (LLINs) are the primary method of malaria prevention. However, the widespread resistance to pyrethroids among major malaria vector species represents a significant threat to the continued efficacy of pyrethroid LLIN. Piperonyl butoxide (PBO) is a synergist that inhibits the activity of metabolic enzymes of the cytochrome P450 family known to detoxify insecticides including pyrethroids. Synergist LLIN incorporating PBO and a pyrethroid may provide improved control compared to pyrethroid-only LLIN.MethodsThe efficacy of VEERALIN® LN (VKA polymers Pvt Ltd, India), an alpha-cypermethrin PBO synergist net was evaluated in experimental huts in M'bé, central Côte d'Ivoire against wild pyrethroid resistant Anopheles gambiae s.s. Comparison was made with a standard alpha-cypermethrin-treated net (MAGNet® LN, VKA polymers Pvt Ltd, India). Nets were tested unwashed and after 20 standardized washes.ResultsVEERALIN® LN demonstrated improved efficacy compared to MAGNet® LN against wild free-flying pyrethroid-resistant An. gambiae s.s. Before washing, VEERALIN® LN produced mortality of An. gambiae s.s. (51%) significantly higher than the standard pyrethroid-only net (29%) (P?<?0.0001). Although there was a significant reduction in mortality with both LLINs after 20 washes, VEERALIN® LN remained superior in efficacy to MAGNet® LN (38 vs 17%) (P?<?0.0001). Blood-feeding was significantly inhibited with both types of insecticide-treated nets relative to the untreated control net (P?<?0.0001). Unwashed VEERALIN® LN induced significantly higher blood-feeding inhibition of An. gambiae s.s. (62.6%) compared to MAGNet® LN (35.4%) (P?<?0.001). The difference persisted after washing, as there was no indication that either LLIN lost protection against biting or blood-feeding. The level of personal protection derived from the use of VEERALIN® LN was high (87%) compared to MAGNet® LN (66-69%) whether unwashed or washed. The AI content of VEERALIN® LN after 20 washes decreased from 6.75 to 6.03 g/kg for alpha-cypermethrin and from 2.95 to 2.64 g/kg for PBO, corresponding to an overall retention of 89% for each compound.ConclusionsThe addition of the synergist PBO to pyrethroid net greatly improved protection and control of pyrethroid-resistant An. gambiae s.s. The pyrethroid-PBO VEERALIN® LN has the potential to reduce transmission in areas compromised by pyrethroid resistance.

Project description:BackgroundPyrethroid resistance in vectors could limit the efficacy of long-lasting insecticidal nets (LLINs) because all LLINs are currently treated with pyrethroids. The goal of this study was to evaluate the efficacy and wash resistance of PermaNet® 3.0 compared to PermaNet® 2.0 in an area of high pyrethroid in Côte d'Ivoire. PermaNet® 3.0 is impregnated with deltamethrin at 85 mg/m2 on the sides of the net and with deltamethrin and piperonyl butoxide on the roof. PermaNet® 2.0 is impregnated with deltamethrin at 55 mg/m2 across the entire net.MethodsThe study was conducted in the station of Yaokoffikro, in central Côte d'Ivoire. The efficacy of intact unwashed and washed LLINs was compared over a 12-week period with a conventionally-treated net (CTN) washed to just before exhaustion. WHO cone bioassays were performed on sub-sections of the nets, using wild-resistant An. gambiae and Kisumu strains. Mosquitoes were collected five days per week and were identified to genus and species level and classified as dead or alive, then unfed or blood-fed.ResultsMortality rates of over 80% from cone bioassays with wild-caught pyrethroid-resistant An. gambiae s.s were recorded only with unwashed PermaNet® 3.0. Over 12 weeks, a total of 7,291 mosquitoes were collected. There were significantly more An. gambiae s.s. and Culex spp. caught in control huts than with other treatments (P < 0.001). The proportion of mosquitoes exiting the huts was significantly lower with the control than for the treatment arms (P < 0.001). Mortality rates with resistant An. gambiae s.s and Culex spp, were lower for the control than for other treatments (P < 0.001), which did not differ (P > 0.05) except for unwashed PermaNet® 3.0 (P < 0.001), which gave significantly higher mortality (P < 0.001).ConclusionsThis study showed that unwashed PermaNet® 3.0 caused significantly higher mortality against pyrethroid resistant An. gambiae s.s and Culex spp than PermaNet® 2.0 and the CTN. The increased efficacy with unwashed PermaNet® 3.0 over PermaNet® 2.0 and the CTN was also demonstrated by higher KD and mortality rates (KD > 95% and mortality rate > 80%) in cone bioassays performed with wild pyrethroid-resistant An. gambiae s.s from Yaokoffikro.

Project description:BackgroundLong-lasting insecticidal nets (LLINs) are a vital tool in the fight against malaria vectors. However, their efficacy in the field can be impacted by several factors, including patterns of usage, net age, mosquito resistance and the delayed mortality effect, all of which could influence malaria transmission. We have investigated the effectiveness of the various brands of LLINs available in markets and households in Cameroon on pyrethroid-resistant mosquitoes and assessed their post-exposure effect.MethodsFollowing quality control assessment on a susceptible laboratory mosquito strain, we evaluated the immediate and delayed mortality effects of exposure to LLINs (both newly bough LLINst and used ones collected from households in Elende village, Cameroon, in 2019) using standard WHO cone tests on Anopheles gambiae and Anopheles funestus populations collected from the Centre region of Cameroon. Alive female mosquitoes were genotyped for various resistance markers at different time points post-exposure to evaluate the impact of insecticide resistance on the efficacy of bednets.ResultsThe laboratory-susceptible strain experienced high mortality rates when exposed to all pyrethroid-only brands of purchased nets (Olyset® Net, Super Net, PermaNet® 2.0, Yorkool®, Royal Sentry®) (Mean±SEM: 68.66 ± 8.35% to 93.33 ± 2.90%). However, low mortality was observed among wild An. funestus mosquitoes exposed to the bednets (0 ± 0 to 28 ± 6.7%), indicating a reduced performance of these nets against field mosquitoes. Bednets collected from households also showed reduced efficacy on the laboratory strain (mortality: 19-66%), as well as displaying a significant loss of efficacy against the local wild strains (mortality: 0 ± 0% to 4 ± 2.6% for An. gambiae sensu lato and 0 ± 0% to 8 ± 3.2% for An. funestus). However, compared to the unexposed group, mosquitoes exposed to bednets showed a significantly reduced longevity, indicating that the efficacy of these nets was not completely lost. Mosquitoes with the CYP6P9a-RR and L119F-GSTe2 mutations conferring pyrethroid resistance showed greater longevity after exposure to the Olyset net than their susceptible counterparts, indicating the impact of resistance on bednet efficacy and delayed mortality.ConclusionThese findings show that although standard bednets drastically lose their efficacy against pyrethroid-resistant field mosquitoes, they still are able to induce delayed mortality in exposed populations. The results of this study also provide evidence of the actual impact of resistance on the quality and efficacy of LLINs in use in the community, with mosquitoes carrying the CYP6P9a-RR and L119F-GSTe2 mutations conferring pyrethroid resistance living longer than their susceptible counterparts. These results highlight the need to use new-generation nets that do not rely solely on pyrethroids.

Project description:A single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. The screening is commonly performed using a multiplex Polymerase Chain Reaction (PCR) which, since it is reliant on a single nucleotide polymorphism, can be unreliable. Here we present a reliable and potentially high throughput method for screening An. gambiae for the kdr mutation.A Hot Ligation Oligonucleotide Assay (HOLA) was developed to detect both the East and West African kdr alleles in the homozygous and heterozygous states, and was optimized for use in low-tech developing world laboratories. Results from the HOLA were compared to results from the multiplex PCR for field and laboratory mosquito specimens to provide verification of the robustness and sensitivity of the technique.The HOLA assay, developed for detection of the kdr mutation, gives a bright blue colouration for a positive result whilst negative reactions remain colourless. The results are apparent within a few minutes of adding the final substrate and can be scored by eye. Heterozygotes are scored when a sample gives a positive reaction to the susceptible probe and the kdr probe. The technique uses only basic laboratory equipment and skills and can be carried out by anyone familiar with the Enzyme-linked immunosorbent assay (ELISA) technique. A comparison to the multiplex PCR method showed that the HOLA assay was more reliable, and scoring of the plates was less ambiguous.The method is capable of detecting both the East and West African kdr alleles in the homozygous and heterozygous states from fresh or dried material using several DNA extraction methods. It is more reliable than the traditional PCR method and may be more sensitive for the detection of heterozygotes. It is inexpensive, simple and relatively safe making it suitable for use in resource-poor countries.

Dataset Information

First detection of N1575Y mutation in pyrethroid resistant Anopheles gambiae in Southern Cote d'Ivoire.

Publications

First detection of N1575Y mutation in pyrethroid resistant <i>Anopheles gambiae</i> in Southern Côte d'Ivoire.

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