Time of exposure and assessment influence the mortality induced by insecticides against metabolic resistant mosquitoes.

BACKGROUND: Increasing metabolic resistance in malaria vector mosquitoes resulted in the development of insecticide-treated nets (ITNs) with active ingredients (AI) that target them. Bioassays that accurately measure the mortality induced by these AIs on ITNs are needed. Mosquito metabolic enzyme expression follows a circadian rhythm. Thus, this study assessed (i) influence of the time of day of mosquito exposure and (ii) timing of assessment of mortality post exposure (24 and 72 h) to ITNs against vectors that are susceptible to pyrethroids and those with metabolic and knockdown resistance mechanisms. METHODS: Two cone bioassay experiments were conducted following World Health Organization (WHO) guidelines. Firstly, on ITNs incorporated with 2 g AI/kg of deltamethrin (DM) alone, or combined with 8 g AI/kg piperonyl butoxide (PBO) synergist, during the day (9:00-14:00 h) and repeated in the evening (18:00-20:00 h). This was followed by a confirmatory experiment during the afternoon (12:00-14:00 h) and repeated in the night (22:00-24:00 h) using mosquitoes unexposed or pre-exposed to PBO for 1 h before exposure to DM ITNs. Each net piece was tested with a minimum of eight cones per time (N = 24). The outcome was mortality after 24 h (M24) or 72 h (M72) of holding. RESULTS: The cone bioassays performed using metabolic resistant mosquitoes during the evening showed significantly lower M24 than those performed in the day for DM: odds ratio (OR) 0.14 [95% confidence interval (CI) 0.06-0.30, p < 0.0001] and DM PBO [OR 0.29 (95% CI 0.18-0.49, p < 0.0001). M72 was higher than M24 for metabolic resistant mosquitoes exposed to DM [OR 1.44 (95% CI 1.09-1.88), p = 0.009] and DM PBO [OR 1.82 (95% CI 1.42-2.34), p < 0.0001]. An influence of hour of experiment and time of assessment was not observed for mosquitoes that had knockdown resistance or that were pyrethroid-susceptible. CONCLUSIONS: Time of day of experiment and hour of assessment of delayed mortality after exposure of mosquitoes are important considerations in evaluating insecticides that interact with mosquito metabolism to counter metabolic resistant mosquitoes. This is important when evaluating field-aged ITNs that may have lower concentrations of AI.

Insecticide-treated eave nets and window screens for malaria control in Chalinze district, Tanzania: a study protocol for a household randomised control trial.

BACKGROUND: Long-lasting insecticidal nets (LLINs) have contributed to the reduction of malaria in sub-Saharan Africa, including Tanzania. However, they rely on daily user behaviour and high coverage which is difficult to maintain. Also, insecticide resistance among malaria vector mosquitoes is contributing to reduced efficacy of control tools. To overcome these problems, we propose to evaluate a new tool for house modification, the insecticide-treated eave nets (ITENs) in combination with insecticide-treated window screens (ITWS) incorporated with dual active ingredient (dual AI) for the control of malaria. METHODS: Four hundred and fifty (450) households with intact walls, open eaves without screens or nets on the windows in Chalinze district will be eligible and recruited upon written informed consent. The households will be randomly allocated into two arms: one with ITENs and ITWS installed and the other without. Malaria parasite detection using a quantitative polymerase chain reaction (qPCR) will be conducted shortly after the long rain (June/July, 2022) as the primary outcome and shortly after the short rain (January/February, 2022) as the secondary outcome. Other secondary outcomes include clinical malaria cases, and density of malaria vectors and nuisance after the short rain and long rain. In addition, surveys will be conducted in households with ITENs and ITWS to estimate the intervention's cost during installation, adverse effects one month after installation, and presence, fabric integrity and user acceptance six and twelve months after installation. Bioefficacy and chemical content will be evaluated twelve months after installation. DISCUSSION: ITENs and ITWS have been shown in Kenya to reduce indoor mosquito density. However, it is not known if indoor mosquito density reduction translates into reduction of malaria cases. Data from the study will measure the potential public health value of an additional intervention for malaria control at the household level in areas of mosquito insecticide resistance that does not require daily adherence. TRIAL REGISTRATION: The study is registered on ClinicalTrials.gov .

From the factory to the field: considerations of product characteristics for insecticide-treated net (ITN) bioefficacy testing.

BACKGROUND: Insecticide-treated nets (ITNs) undergo a series of tests to obtain listing by World Health Organization (WHO) Prequalification. These tests characterize the bioefficacy, physical and chemical properties of the ITN. ITN procurers assume that product specifications relate to product performance. Here, ITN test methods and their underlying assumptions are discussed from the perspective of the ITN manufacturing process and product characteristics. METHODS: Data were extracted from WHO Pesticide Evaluation Scheme (WHOPES) meeting reports from 2003 to 2017, supplemented with additional chemical analysis to critically evaluate ITNs bioassays with a focus on sampling, washing and wash resistance, and bioefficacy testing. Production methods for ITNs and their impact on testing outcomes are described. RESULTS AND RECOMMENDATIONS: ITNs are not homogenous products. They vary within panels and between the sides and the roof. Running tests of wash resistance using a before/after tests on the same sample or band within a net reduces test variability. As mosquitoes frequently interact with ITN roofs, additional sampling of the roof when evaluating ITNs is advisable because in nets where roof and sides are of the same material, the contribution of roof sample (20-25%) to the average is less than the tolerance for the specification (25%). Mosquito mortality data cannot be reliably used to evaluate net surface concentration to determine regeneration time (RT) and resistance to washing as nets may regenerate beyond the insecticide concentrations needed to kill 100% of susceptible mosquitoes. Chemical assays to quantify surface concentration are needed. The Wash Resistance Index (WRI) averaged over the first four washes is only informative if the product has a log linear loss rate of insecticide. Using a WRI that excludes the first wash off gives more reliable results. Storage conditions used for product specifications are lower than those encountered under product shipping and storage that may exceed 50 °C, and should be reconsidered. Operational monitoring of new ITNs and linking observed product performance, such as bioefficacy after 2 or 3 years of use, with product characteristics, such as WRI, will aid the development of more robust test methods and product specifications for new products coming to market.

Comparing the un-comparable: Olyset Plus and Olyset, different malaria impact.

BACKGROUND: In a recent article in The Lancet, Protopopoff et al. stated that insecticide resistance must be tackled and concluded that adding the insecticide synergist piperonyl butoxide (PBO) to a bed net with a pyrethroid as principal insecticide might be a part of the response. MAIN TEXT: The study in Tanzania compares malaria prevalence between users of two different nets with the principal insecticide permethrin: Olyset and Olyset Plus (Olyset+), the latter also holding the synergist molecule PBO, the first not. The article is based on randomized cluster trial of very high quality, but Olyset+ exposes much more permethrin at the surface so the higher efficacy may not be because of the added PBO. CONCLUSION: Data published by the World Health Organization (WHO) when evaluating Olyset+ as well of the study of Protopopoff et al. showed that much more permethrin is available on the surface of Olyset+ than on the surface of Olyset and the relatively small and rapidly dwindling dosage of PBO may have nothing to do with the superior effect of Olyset+. The WHO should not change politics for "PBO nets" based on this study alone.

Supplementary effect and durability of prototype insecticide-treated eave curtains on indoor resting mosquitoes in Kadibo division, Western Kenya.

Background: Use of insecticide treated bednets (ITNs) was a breakthrough in the fight against malaria. However, ITNs are only effective when properly used. Recent reports indicate low compliance in ITN usage and changes in biting times of malaria vectors with early and late biting cases recorded when people are not sleeping under their nets. Hence, there is a need to develop methods to supplement or replace the use of ITNs for malaria vector control. A field trial was conducted to investigate the effect and longevity of prototypes of long lasting impregnated UV protected eave nets, curtains and door hangers (fully screened houses), compared to houses with bednets only, in traditional East African houses. Materials and methods: A randomised controlled trial was carried out in the Ahero rice irrigation scheme in Nyando district, Western Kenya. Eighty houses with open eaves were randomly selected. Forty of these houses were fully screened (FSH+LLINs) with long lasting insecticide-treated nets/curtains used to screen the eaves, windows and doors. The FSH materials were produced with anti-UV additives. The other 40 houses served as controls. Long lasting insecticide-treated bednets (LLINs) were suspended over all sleeping areas in the control and intervention houses. Indoor resting Anopheles mosquitoes were collected using pyrethrum spray catches (PSC) during both dry and wet seasons. Indoor population densities of anophelines were compared between intervention (FSH+LLINs) and control (LLINs) houses. Loss of insecticide (deltamethrin) was compared after 12 and 24 months for both the FSH materials and LLINs through bioassays and chemical analyses. Results: In the FSH+LLINs houses densities of indoor resting Anopheles funestus and An. arabiensis were reduced by 82% (RR=0.18, 95% CI 0.09-0.36, P<0.0001) and 70% (RR=0.30, 95% CI 0.15-0.58, P<0.0001), respectively. No significant difference was recorded for indoor resting Culex spp. (RR=0.95, 95% CI 0.48-1.86, P=0.8). The population of indoor resting bloodfed An. arabiensis and An. funestus was reduced by 72% (RR=0.22, 95% CI 0.09-0.51, P<0.0001) and 84% (RR=0.16, 95% CI 0.07-0.33, P<0.0001) in the FSH+LLINs houses and LLIN houses, respectively. Insecticide loss in eave nets did not depend on the side of the house where the nets were placed. The eave nets showed little loss of bio efficacy over the 12-24 months period. Conclusions: The study revealed that the use of insecticide-treated nets on the eaves and windows combined with door hangers largely impeded entrance of anopheline mosquitoes into houses and can be used to compliment LLINs for household protection. The eave nets were suspended from wood structures near the eave and remained in place when walls were re-plastered. The nets are therefore not depending on daily compliance behaviour and provide protection for the entire household.

Cost of integrated vector control with improved sanitation and road infrastructure coupled with the use of slow-release Bacillus sphaericus granules in a tropical urban setting.

A field test of integrated vector control was conducted in a tropical urban setting with a combination of a floating, slow-release, granular formulation of Bacillus sphaericus and environmental engineering measures (renovation of roads, collective water pumps, and cesspool lids). The targets were Culex quinquefasciatus and Anopheles gambiae in the two biggest towns of Burkina Faso (West Africa). Within the intervention zone, water pumping stations were improved and the surroundings drained to prevent the accumulation of stagnant water. Roads were leveled and given either simple gutters on each side or a concrete channel on one side to drain runoff water. Garbage containers were installed to provide an alternative to the drainage channels for waste disposal. Septic tanks were modified so that they could be emptied without destroying their lid. This study showed that it is possible to implement mosquito control in a tropical urban environment with teams of young people rapidly trained to apply a biological larvicide without any tools other than an iron bar to lift cesspool lids. Environmental improvements were initially costly, but demanded little subsequent expenditure. Local inhabitants' committees were mobilized to provide people with information and monitor the efficacy of the measures. Compared with what people spent individually on mosquito prevention and malaria medicine, these measures were not expensive, but many expected the community to pay for them from existing taxes, e.g., for water treatment and disposal. The necessary funding and logistics require a municipal organization with neighborhood support, if the measures are to be effective.

Strength of bed nets as function of denier, knitting pattern, texturizing and polymer.

BACKGROUND: Bursting strength is a standard method for evaluating mosquito net strength. This article suggests that tension strength with one grab and one hook better represent how holes are generated in bed nets in real life. METHODS: Measurements of bursting strength and tension strengths in the two directions are analysed for eight model nets created for the study. The nets were made in the most commonly used denier (75 and 100 D) and mesh (156 holes/inch(2)) for multifilament polyester yarns, texturized or not, and with 4 or 6 sided holes. All were made from one polyester quality. Data was arranged in a randomized, complete block design and analysed for significant variables and their interactions. Data was then subjected to regression analyses using net square metre weight as a weighting factor with stepwise removal of variables. This revealed how the four textile variables interacted and allowed for making predictions for the strength of commercial nets in polyester or polyethylene. RESULTS: For the model nets, higher denier provided higher bursting strength and tension strengths, texturizing weakened nets and four-sided holes were stronger than six-sided holes. Even when compensating for square metre weight, 100 D nets are stronger than 75 D nets. Results for the commercial polyester net nets are less clear, probably because of different qualities of polyester. Tensile strength: a 75 denier net knitted tightly to provide the same square metre weight as a standard 100 denier net therefore does not obtain the same strength. Polyethylene nets are made of mono-fibre yarns and, therefore, have higher tension strength in both directions than multifilament polyester nets. For bursting strength results overlap for 100 denier yarns of both yarn types. As a class, commercial polyethylene nets are stronger than commercial polyester net whatever method is used for evaluation. CONCLUSION: Tension strength measured in the length and width directions of the net using one hook and one clamp provide new relevant data as this method more closely imitates the cause of tear holes in nets as they occur in real life use. Using this methodology, the commercial monofilament yarn polyethylene nets are significantly stronger than the commercial multifilament polyester nets. This test method should be applied for nets used for years in the field.

Decreased motivation in the use of insecticide-treated nets in a malaria endemic area in Burkina Faso.

BACKGROUND: The use of insecticide-treated nets (ITN) is an important tool in the Roll Back Malaria (RBM) strategy. For ITNs to be effective they need to be used correctly. Previous studies have shown that many factors, such as wealth, access to health care, education, ethnicity and gender, determine the ownership and use of ITNs. Some studies showed that free distribution and public awareness campaigns increased the rate of use. However, there have been no evaluations of the short- and long-term impact of such motivation campaigns. A study carried out in a malaria endemic area in south-western Burkina Faso indicated that this increased use declined after several months. The reasons were a combination of the community representation of malaria, the perception of the effectiveness and usefulness of ITNs and also the manner in which households are organized by day and by night. METHODS: PermaNet 2.0 and Olyset were distributed in 455 compounds at the beginning of the rainy season. The community was educated on the effectiveness of nets in reducing malaria and on how to use them. To assess motivation, qualitative tools were used: one hundred people were interviewed, two hundred houses were observed directly and two houses were monitored monthly throughout one year. RESULTS: The motivation for the use of bednets decreased after less than a year. Inhabitants' conception of malaria and the inconvenience of using bednets in small houses were the major reasons. Acceptance that ITNs were useful in reducing malaria was moderated by the fact that mosquitoes were considered to be only one of several factors which caused malaria. The appropriate and routine use of ITNs was adversely affected by the functional organization of the houses, which changed as between day and night. Bednets were not used when the perceived benefits of reduction in mosquito nuisance and of malaria were considered not to be worth the inconvenience of daily use. CONCLUSION: In order to bridge the gap between possession and use of bednets, concerted efforts are required to change behaviour by providing accurate information, most particularly by convincing people that mosquitoes are the only source of malaria, whilst recognising that there are other diseases with similar symptoms, caused in other ways. The medical message must underline the seriousness of malaria and the presence of the malaria vector in the dry season as well as the wet, in order to encourage the use of bednets whenever transmission can occur. Communities would benefit from impregnated bednets and other vector control measures being better adapted to their homes, thus reducing the inconvenience of their use.

Impact of slow-release Bacillus sphaericus granules on mosquito populations followed in a tropical urban environment.

A floating, slow-release, granular formulation of Bacillus sphaericus (Neide) was used to control mosquito larvae in two suburban areas of two tropical cities: Ouagadougou and Bobo-Dioulasso, Burkina Faso. A circular area of 2 km2, diameter 1,600 m, was treated in each city using a similar, smaller area 1 km away as an untreated control. Mosquito captures were made in houses in four concentric circles, from the periphery to the center; each circle was 50 m in width. Mosquitoes were captured in CDC light traps or from human landings. More than 95% of the mosquitoes were Culex quinquefasciatus (Say) (Diptera: Culicidae). The human landing catches provided twice as many mosquitoes as did the CDC traps/night/house. The treatments resulted in important reductions relative to the control area and to preintervention captures. The reduction was more prominent in the inner circle (up to 90%) than in the outer circle (50-70%), presumably because of the impact of immigrating mosquitoes from nontreated breeding sites around the intervention area. This effect was more pronounced for light trap catches than from human landings. The impact of treatment was also measured as the mean ratio of mosquito density in the two outer circles to that of the two inner circles. This ratio was approximately 1:1 before the intervention and reached 1:0.43 during the intervention. This comparison does not depend on the assumption that, in the absence of intervention, the mosquito population development in the two areas would have been identical, but does depend on the homogeneity of the intervention area. The study showed that it is possible to organize mosquito control in a tropical, urban environment by forming and rapidly training teams of young people to carry out the mosquito control mostly using a biopesticide that can be applied without any tools except an iron bar to lift lids on some cesspits.

Median knock-down time as a new method for evaluating insecticide-treated textiles for mosquito control.

BACKGROUND: Insecticide treated bed nets are major tools for the Roll Back Malaria campaign. There are two types of Long-Lasting Insecticide-treated Nets (LNs) on the market: coated nets and insecticide-incorporated nets. Nets provided to this market need a recommendation from the World Health Organization to be purchased by donors and NGOs. During laboratory study (phase I), the first step consists in evaluating the wash resistance of a new LN product. When insecticide-incorporated nets are washed, it takes time to regenerate the insecticidal activity, i.e. insecticide must migrate to the net surface to be accessible to mosquitoes. The interval of time required for regeneration must be carefully determined to ensure the accuracy of further results. WHOPES procedures currently recommend the determination of the regeneration time by using mortality data. However, as mortality cannot exceed 100%, a LN that regenerates a surface concentration exceeding the dosage for 100% mortality, will have its regeneration time underestimated. METHODS: The Median Knock Down Time (MKDT) was determined as function of insecticide dosage on an inert surface, glass, and on polyester nettings using an acetone solution or a simple emulsion. Dosage response was also established for mortality data. The same method was then applied to a commercially polyethylene netting, currently under WHOPES evaluation, to determine the dynamics of regeneration as function of repeated washings. The deltamethrin content of these nets was estimated by Capillary Gas Chromatography (GC-ECD). RESULTS: MKDT was a linear function of log insecticide dosage on glass as on nettings. Mortality data were either 0 or 100% for most concentrations except for a narrow range. MKDT was log linear function of total deltamethrin content in a commercial polyethylene net exposed to washings. The regeneration time of this net increased with the number of washes and MKDT became higher. A new, easy and rapid method to determine MKDT is suggested. DISCUSSION: The MKDT is linearly correlated to log dosage on a given substrate and shows no saturation as mortality data do. It is suited to determine regeneration time of a product that is exposed to a stress, like washing or heating, where the process impacts on the bio-availability of the insecticide. Mortality data are useful for measuring product efficacy, whereas MKDT are better to measure dynamics of surface concentration like regeneration after a stressing process. Change in MKDT can be used to illustrate the loss of insecticide due to washing, but the slope of the curve is product and surface-dependent.

Microbial control in Southeast Asia.

Beginning in the 1980s, concerns about the deleterious effects of synthetic pesticides have driven a significant Southeast Asian research and development effort directed towards alternative pest control strategies, including the use of microbial control agents. Despite this effort, use of microbial control agents has grown slowly in the region. This is the result of an interplay between internal factors such as economics, national research programs, farmer education, manufacturing capabilities and regulatory frameworks, and external factors such as the influence of neighboring countries (particularly China), the availability of competitive pest control products, import regulations on pesticide residues and the activities of donor agencies. The role of these factors in providing both incentives and barriers to the adoption of microbial control are explored, and examples of promising projects are examined as a means of pointing the way forward towards increased progress in the future.

Personal protection of long lasting insecticide-treated nets in areas of Anopheles gambiae s.s. resistance to pyrethroids.

BACKGROUND: The development of mosquito nets pre-treated with insecticide, Long Lasting Impregnated Nets (LLINs) that last the life span of the net, is a solution to the difficulty of the re-impregnation of conventional nets. Even if they showed a good efficacy in control conditions, their efficacy in the field, particularly in areas with resistance of Anopheles gambiae to pyrethroids, is not well documented. This study compares wide (Olyset) and small (Permanet) mesh LLINs in field conditions, using entomological parameters. METHODS: The two LLINs were tested in a rice-growing area of south-western Burkina Faso (West Africa) with year around high density of the main malaria vector An. gambiae s.s. In the study village (VK6), there is a mixed population of two molecular forms of An. gambiae, the S-form which dominates during the rainy season and the M-form which dominates the rest of the year. The two LLINs Olyset and Permanet were distributed in the village and 20 matched houses were selected for comparison with four houses without treated nets. RESULTS: Mosquito entrance rate was ten fold higher in control houses than in houses with LLINs and there was no difference between the two net types. Among mosquitoes found in the houses, 36 % were dead in LLIN houses compared to 0% in control houses. Blood feeding rate was 80 % in control houses compared to 43 % in LLIN houses. The type of net did not significantly impact any of these parameters. No mosquitoes were found inside Permanet, whereas dead or dying mosquitoes were collected inside the Olyset. More than 60% of mosquitoes found on top or inside the nets had had blood meals from cattle, as shown by ELISA analysis. CONCLUSION: The percentage of blood-fed mosquitoes in a bed net study does not necessarily determine net success. The efficacy of the two types of LLINs was comparable, during a period when the S-form of An. gambiae was carrying the kdr gene. Significantly higher numbers of mosquitoes were collected in control houses compared to intervention houses, indicating that the LLINs provided an additional deterrent effect, which enhanced their expected prevention capacity.

Prevention of mosquito nuisance among urban populations in Burkina Faso.

This paper addresses the problems of mosquito control in urban areas of Burkina Faso. The main objectives are to examine relevant socio-cultural aspects in relation to a mosquito control intervention using a biolarvicide with main emphasis on local perceptions of mosquito nuisance and existing practices of mosquito control, including the cost of protective measures at household level. This is the report of an inter-disciplinary research project carried out in the two major towns of Burkina Faso, Bobo-Dioulasso and Ouagadougou, in 1999 and 2000, respectively. Both quantitative and qualitative methods were used in the ethnographic part of the study. Two questionnaire surveys were conducted in both study areas: one prior to the intervention (n=1083) and the other after the intervention of the treatments with bio-larvicide (n=956). In addition, 70 in-depth interviews and 17 focus group discussions (FGDs) were conducted. The findings show that mosquitoes are considered an important problem in the urban areas, both as a nuisance and a health risk and that the local population is very active in applying mosquito control measures at the household level. The intervention project was received positively by the local population with a decline in the perceived level of annoyance. The causal relationship between mosquitoes and malaria is clear, but the explanatory framework of the relationship between mosquitoes and other diseases is still under debate. The most common prevention methods are mosquito coils and aerosol spray, even though bed nets are perceived to be the most efficient and effective method. The investments in coils and aerosol sprays alone would mean an increase of 40% in the national figures for health expenditure at household level.