Bendiocarb, a potential alternative against pyrethroid resistant Anopheles gambiae in Benin, West Africa

<p>Abstract</p> <p>Background</p> <p><it>Anopheles gambiae</it>, the main malaria vector in Benin has developed high level of resistance to pyrethroid insecticides, which is a serious concern to the future use of long-lasting insecticidal nets (LLIN) and indoor residual spraying (IRS). In this context, one of the pathways available for malaria vector control would be to investigate alternative classes of insecticides with different mode of action than that of pyrethroids. The goal of this study was to evaluate under field conditions the efficacy of a carbamate (bendiocarb) and an organophosphate (fenitrothion) against pyrethroid-resistant <it>An. gambiae s.s</it>.</p> <p>Methods</p> <p>Wild populations and females from laboratory colonies of five days old <it>An. gambiae </it>were bio-assayed during this study. Two pyrethroids (deltamethrin and alphacypermethrin), an organophosphate (fenitrothion), a carbamate (bendiocarb) and a mixture of an organophosphate (chlorpyriphos + a pyrethroid deltamethrin) were compared in experimental huts as IRS treatments. Insecticides were applied in the huts using a hand-operated compression sprayer. The deterrency, exophily, blood feeding rate and mortality induced by these insecticides against <it>An. gambiae </it>were compared to the untreated control huts.</p> <p>Results</p> <p>Deltamethrin, alphacypermethrin and bendiocarb treatment significantly reduced mosquito entry into the huts (p < 0.05) compared to untreated huts. Blood feeding rates in huts treated with fenitrothion and the mixture chlorpyriphos/deltamethrin were reduced from 10.95% respectively to 3.7% and 4.47% three months after treatment and from 10.20% to 4.4% and 2.04% four months after treatment. Exophily rates in huts with deltamethrin, alphacypermethrin and the mixture chlorpyriphos/deltamethrin were significantly higher than in the huts with fenitrothion. Deltamethrin and alphacypermethrin had the lowest mortality rate while fenitrothion killed 100% of <it>An. gambiae </it>(in the first month) and 77.8% (in the fourth month). Bendiocarb and the mixture chlorpyriphos/deltamethrin mortality rates ranged from 97.9 to 100% the first month and 77.7-88% the third month respectively.</p> <p>Conclusion</p> <p>After four months, fenitrothion, bendiocarb and the mixture chlorpyriphos/deltamethrin performed effectively against pyrethroid-resistant <it>Anopheles</it>. These results showed that bendiocarb could be recommended as an effective insecticide for use in IRS operations in Benin, particularly as the mixture chlorpyriphos/deltamethrin does not have WHOPES authorization and complaints were mentioned by the sleepers about the safety and smell of fenitrothion.</p

Knowledge-Attitudes-Practices About Malaria Among Communities in Southern Benin

Malaria still remains the main public health problem in Benin. We explored the determinants that influenced malaria treatment as well as protective behaviors, to generate a framework of useful ideas as alternative strategies against malaria. A cross-sectional survey of the knowledge, attitudes and practices (KAP) was conducted at Hozin, Vakon and Agblangandan districts in southern region of Benin. Descriptive statistics were computed and mixed logistic regression helped evaluating the relationship between frequency of each category of severity of malaria and sex group, educational level, treatment, means of self-protection against mosquitoes and identification of the cause of malaria. A significant proportion 750 (81.3%) (p&lt;0.001) of participants stated that malaria was caused by mosquitoes. The respondents who mentioned sun as the cause of malaria, have trivialized more malaria in a proportion of about 59.30% (OR=2.67 [95% CI 1.61-4.44]) followed by those who have reported the cause of body weakness (43.68%) (OR=2.97 [95% CI 1.68-5.28]). Poor knowledge justifies the trivialization of the disease and poor management of malaria control means. National Malaria Control Programs should improve access to education, especially for women and could help improving prevention and control behaviours against malaria in communities

Impact of three years of large scale Indoor Residual Spraying (IRS) and Insecticide Treated Nets (ITNs) interventions on insecticide resistance in Anopheles gambiae s.l. in Benin

<p>Abstract</p> <p>Background</p> <p>In Benin, Indoor Residual Spraying (IRS) and long-lasting insecticidal nets (LLINs) are the cornerstones of malaria prevention. In the context of high resistance of <it>Anopheles gambiae </it>to pyrethroids, The National Malaria Control Program (NMCP) has undertaken a full coverage of IRS in a no-flood zone in the Oueme region, coupled with the distribution of LLINs in a flood zone. We assessed the impact of this campaign on phenotypic resistance, <it>kdr </it>(knock-down resistance) and <it>ace-1^R</it>(insensitive acetylcholinesterase) mutations.</p> <p>Methods</p> <p>Insecticides used for malaria vector control interventions were bendiocarb WP (0.4 g/m²) and deltamethrin (55 mg/m²), respectively for IRS and LLINs. Susceptibility status of <it>An. gambiae </it>was assessed using World Health Organization bioassay tests to DDT, permethrin, deltamethrin and bendiocarb in the Oueme region before intervention (2007) and after interventions in 2008 and 2010. <it>An. gambiae </it>specimens were screened for identification of species, molecular M and S forms and for the detection of the West African <it>kdr </it>(L1014F) as well as <it>ace-1^R</it>mutations using PCR techniques.</p> <p>Results</p> <p>The univariate logistic regression performed showed that <it>kdr </it>frequency has increased significantly during the three years in the intervention area and in the control area. Several factors (LLINs, IRS, mosquito coils, aerosols, use of pesticides for crop protection) could explain the selection of individual resistant <it>An. gambiae</it>. The <it>Kdr </it>resistance gene could not be the only mechanism of resistance observed in the Oueme region. The high susceptibility to bendiocarb is in agreement with a previous study conducted in Benin. However, the occurrence of <it>ace-1^R</it>heterozygous individuals even on sites far from IRS areas, suggests other factors may contribute to the selection of resistance other than those exerted by the vector control program.</p> <p>Conclusion</p> <p>The results of this study have confirmed that <it>An.gambiae </it>have maintained and developed the resistance to pyrethroids, but are still susceptible to bendiocarb. Our data clearly shows that selection of resistant individuals was caused by other insecticides than those used by the IRS and LLINs.</p

Analysis-ready datasets for insecticide resistance phenotype and genotype frequency in African malaria vectors

The impact of insecticide resistance in malaria vectors is poorly understood and quantified. Here a series of geospatial datasets for insecticide resistance in malaria vectors are provided, so that trends in resistance in time and space can be quantified, and the impact of resistance found in wild populations on malaria transmission in Africa can be assessed. Specifically, data have been collated and geopositioned for the prevalence of insecticide resistance, as measured by standard bioassays, in representative samples of individual species or species complexes. Data are provided for the Anopheles gambiae species complex, the Anopheles funestus subgroup, and for nine individual vector species. Data are also given for common genetic markers of resistance to support analyses of whether these markers can improve the ability to monitor resistance in low resource settings. Allele frequencies for known resistance-associated markers in the Voltage-gated sodium channel (Vgsc) are provided. In total, eight analysis-ready, standardised, geopositioned datasets encompassing over 20,000 African mosquito collections between 1957 and 2017 are released

Resistance status of Anopheles gambiae s.l. to insecticides following the 2011 mass distribution campaign of long-lasting insecticidal nets (LLINs) in the Plateau Department, south-eastern Benin

Abstract Background In 2011, Benin’s National Malaria Control Programme (NMCP) organized a nationwide mass distribution campaign of LLINs throughout the country. Following this intervention, it was important to assess whether the level of susceptibility of malaria vectors to insecticides had remained the same as compared to the pre-intervention period. The current study investigated this. Methods Larval collections were conducted in Ifangni, Sakété, Pobè and Kétou districts located in Plateau department, Southeastern Benin before (2009) and after (2012–2013) LLIN distribution. Anopheles gambiae sensu lato (s.l.) larvae from the 4 study districts were reared to adulthood and WHO susceptibility tests were conducted. The insecticides tested were deltamethrin (0.05%), permethrin (0.75%), bendiocarb (0.1%) and DDT (4%). Molecular species identification as well as, the characterization of the kdr L1014F mutation were also performed in the An. gambiae s.l. complex using PCR method. Results Overall, a significant decrease in mortality rates of An. gambiae s.l. to deltamethrin (0.05%), permethrin (0.75%) and DDT (4%) was observed post-LLIN distribution, respectively: (100% vs 80.9%, p &lt; 0.0001), (77.5% vs 70%, p = 0.01) and, (47.8% vs 4.4%, p &lt; 0.0001). By contrast, susceptibility of vectors to bendiocarb (0.1%) remained the same (100% mortality in the WHO susceptibility tube tests) pre- and post-intervention. An increase in the kdr L1014F frequency was observed post-LLIN distribution [F(kdr) = 0.91)] compared to the pre-intervention period [F(kdr) = 0.56], p &lt; 0.0001. Anopheles coluzzii and An. gambiae were the two molecular species identified in the study area. Conclusion The decrease susceptibility to pyrethroids and DDT as well as, the increase in the frequency of the kdr L1014F mutation after the intervention stressed at the time, the need for the development and implementation of effective insecticide resistance management strategies. At present, an update of the vectors resistance status in the area is also necessary for decision-making. </jats:sec

Insecticide resistance status in Anopheles gambiae in southern Benin

BACKGROUND: The emergence of pyrethroid resistance in Anopheles gambiae has become a serious concern to the future success of malaria control. In Benin, the National Malaria Control Programme has recently planned to scaling up long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) for malaria prevention. It is, therefore, crucial to monitor the level and type of insecticide resistance in An. gambiae, particularly in southern Benin where reduced efficacy of insecticide-treated nets (ITNs) and IRS has previously been reported. METHODS: The protocol was based on mosquito collection during both dry and rainy seasons across forty districts selected in southern Benin. Bioassay were performed on adults collected from the field to assess the susceptibility of malaria vectors to insecticide-impregnated papers (permethrin 0.75%, delthamethrin 0.05%, DDT 4%, and bendiocarb 0.1%) following WHOPES guidelines. The species within An. gambiae complex, molecular form and presence of kdr and ace-1 mutations were determined by PCR. RESULTS: Strong resistance to permethrin and DDT was found in An. gambiae populations from southern Benin, except in Aglangandan where mosquitoes were fully susceptible (mortality 100%) to all insecticides tested. PCR showed the presence of two sub-species of An. gambiae, namely An. gambiae s.s, and Anopheles melas, with a predominance for An. gambiae s.s (98%). The molecular M form of An. gambiae was predominant in southern Benin (97%). The kdr mutation was detected in all districts at various frequency (1% to 95%) whereas the Ace-1 mutation was found at a very low frequency (<or= 5%). CONCLUSION: This study showed a widespread resistance to permethrin in An. gambiae populations from southern Benin, with a significant increase of kdr frequency compared to what was observed previously in Benin. The low frequency of Ace-1 recorded in all populations is encouraging for the use of bendiocarb as an alternative insecticide to pyrethroids for IRS in Benin

Field Durability of &amp;lt;i&amp;gt;Yorkool&amp;lt;sup&amp;gt;&amp;#174;&amp;lt;/sup&amp;gt;LN&amp;lt;/i&amp;gt; Nets in the Benin Republic

CONTEXT: Recent publications on WHO recommended methods for estimating the survival of LLINs are good guidelines for assessing the performance of long-lasting insecticidal nets (LLINs). Thus, this field trial study was undertaken to evaluate the durability of the Yorkool® LN mosquito net distributed during the 2017 campaign in Benin. METHODS: The monitoring of Yorkool® LN nets was carried out in two districts (Djougou III and Barienou) in Djougou, department of Donga, northern Benin from October 2017 to March 2019. A representative sample of 250 households that had received the Yorkool® LN polyester LLINs during the 2017 campaign was selected in the rural and urban areas of each district and monitored for 6, 12 and 18 months. An evaluation of the survival of Yorkool® LN nets was conducted based on the rate of loss and physical condition of the surviving nets as measured by the proportional hole index (pHI). Finally, the chemical efficacy of these LLINs during each period was determined using the WHO cone tests. RESULTS: Survival of Yorkool® LN nets was similar in both rural and urban areas, although there was a difference in survival between the 6-month (95.3%), 12-month (89.7%), and 18-month follow-up periods (74.4%). A difference in survival was also observed between the NetCalc model (84%) compared to the Yorkool® LN nets of this study (74.4%). The attrition rate was 29.6% for LLINs at 18 months. Surprisingly, the physical integrity of the LLINs was minimally affected in the municipality. Indeed, the proportion of mosquito nets in good condition without a hole was 51.8% compared to 56.8% with a hole after 18 months. Only 7.8% of the LLINs in the two districts were damaged compared to 2.6% which needed to be replaced. The washing frequency, location of the LLINs and the frequency of use are some factors contributing to the appearance of the holes in LLINs. The bio-efficacy results of LLINs based on the cone test were good with mortality rates of 74%, 66%, 72% and 58% respectively after baseline, 6, 12 and 18 months of use. CONCLUSIONS: The observed differences in the survival of Yorkool® LN nets are due to community living conditions and movements and not to the equipment used to manufacture LLINs. However, the estimated median survival has shown that Yorkool® LN nets would have an average lifespan of 2 years 8 months despite their fairly good physical condition. These results may be useful to the National Malaria Control Program (NMCP) during the period of replacement of these nets on the field

Efficacy of Pyrethroid-Pyriproxyfen and Pyrethroid-Chlorfenapyr Long-Lasting Insecticidal Nets (LLINs) for the Control of Non-Anopheles Mosquitoes: Secondary Analysis from a Cluster Randomised Controlled Trial (cRCT)

The efficacy of a vector control tool in reducing mosquito biting is crucial for its acceptability. The present study compared the vector density of Culex spp. And Mansonia spp. across clusters, which received two dual-active ingredient (a.i.) long-lasting insecticidal nets (LLINs) and a standard pyrethroid-only LLIN, and assessed the seasonality of these mosquito genera. A total of 85,723 Culex spp. and 144,025 Mansonia spp. were caught over the study period. The density of Culex and Mansonia was reduced in all three arms over the study period. There was no evidence of a significant reduction in the indoor or outdoor density of Culex spp. in either dual-a.i. LLIN arm as compared to the standard pyrethroid-only net arm. A similar trend was observed with Mansonia spp. A high density of Culex spp. was found both in rainy and dry seasons, while for Mansonia spp., this was mainly observed during the rainy season. These results suggest that the novel insecticides in the dual-a.i. LLINs did not have an additional impact on these species and that pyrethroids might still be effective on them. Further work is required to determine whether these species of mosquitoes have resistance to the insecticides tested in this trial

Cotton pest management practices and the selection of pyrethroid resistance in Anopheles gambiae population in Northern Benin

<p>Abstract</p> <p>Background</p> <p>Pyrethroid insecticides, carbamate and organophosphate are the classes of insecticides commonly used in agriculture for crop protection in Benin. Pyrethroids remain the only class of insecticides recommended by the WHO for impregnation of bed nets. Unfortunately, the high level of pyrethroid resistance in <it>Anopheles gambiae </it>s.l., threatens to undermine the success of pyrethroid treated nets. This study focuses on the investigation of agricultural practices in cotton growing areas, and their direct impact on larval populations of <it>An. gambiae </it>in surrounding breeding sites.</p> <p>Methods</p> <p>The protocol was based on the collection of agro-sociological data where farmers were subjected to semi-structured questionnaires based on the strategies used for crop protection. This was complemented by bioassay tests to assess the susceptibility of malaria vectors to various insecticides. Molecular analysis was performed to characterize the resistance genes and the molecular forms of <it>An. gambiae</it>. Insecticide residues in soil samples from breeding sites were investigated to determine major factors that can inhibit the normal growth of mosquito larvae by exposing susceptible and resistant laboratory strains.</p> <p>Results</p> <p>There is a common use by local farmers of mineral fertilizer NPK at 200 kg/ha and urea at 50 kg/hectare following insecticide treatments in both the Calendar Control Program (CCP) and the Targeted Intermittent Control Program (TICP). By contrast, no chemicals are involved in Biological Program (BP) where farmers use organic and natural fertilizers which include animal excreta.</p> <p>Susceptibility test results confirmed a high resistance to DDT. Mean mortality of <it>An. gambiae </it>collected from the farms practicing CCP, TICP and BP methods were 33%, 42% and 65% respectively. <it>An. gambiae </it>populations from areas using the CCP and TICP programs showed resistance to permethrin with mortality of 50% and 58% respectively. By contrast, bioassay test results of <it>An. gambiae </it>from BP areas gave a high level of susceptibility to permethrin with an average mortality of 94%.</p> <p>Molecular analysis identified <it>An. gambiae </it>s.s, and <it>An. arabiensis </it>with a high predominance of <it>An. gambiae s.s </it>(90%). The two molecular forms, M and S, were also determined with a high frequency of the S form (96%).</p> <p>The <it>Kdr </it>gene seemed the main target- site resistance mechanism detected in CCP, TICP, and BP areas at the rates ranging from 32 to 78%. The frequency of <it>ace-1R </it>gene was very low (< 0.1).</p> <p>The presence of inhibiting factors in soil samples under insecticide treatments were found and affected negatively in delaying the development of <it>An. gambiae </it>larval populations.</p> <p>Conclusions</p> <p>This research shows that <it>Kdr </it>has spread widely in <it>An. gambiae</it>, mainly in CCP and TICP areas where pyrethroids are extensively used. To reduce the negative impact of pesticides use in cotton crop protection, the application of BP-like programs, which do not appear to select for vector resistance would be useful. These results could serve as scientific evidence of the spread of resistance due to a massive agricultural use of insecticides and contribute to the management of pesticides usage on cotton crops hence reducing the selection pressure of insecticides on <it>An. gambiae </it>populations.</p