Review of Issues on Residual Malaria Transmission.

Residual malaria transmission is the actual maintained inoculation of Plasmodium, in spite of a well-designed and implemented vector control programs, and is of great concern for malaria elimination. Residual malaria transmission occurs under several possible circumstances, among which the presence of exophilic vector species, such as Anopheles dirus, or indoor- and outdoor-biting vectors, such as Anopheles nili, or specific behavior, such as feeding on humans indoors, then resting or leaving the house the same night (such as Anopheles moucheti) or also changes in behavior induced by insecticides applied inside houses, such as the well-known deterrent effect of permethrin-treated nets or the irritant effect of DDT. The use of insecticides may change the composition of local Anopheles populations, such as A. arabiensis taking up the place of A. gambiae in Senegal, A. aquasalis replacing A. darlingi in Guyana, or A. harrisoni superseding A. minimus in Vietnam. The change in behavior, such as biting activity earlier than usually reported-for example, Anopheles funestus after a large-scale distribution of long-lasting insecticidal nets-or insecticide resistance, in particular the current spread of pyrethroid resistance, could hamper the efficacy of classic pyrethroid-treated long-lasting insecticidal nets and maintained transmission. These issues must be well documented in every situation to elaborate, implement, monitor, and evaluate tailored vector control programs, keeping in mind that they must be conceived as integrated programs with several well and appropriately coordinated approaches, combining entomological but also parasitological, clinical, and social methods and analyses. A successful integrated vector control program must then be designed to reduce transmission and incidence rates of malaria morbidity and overall mortality.

Coalition: Advocacy for prospective clinical trials to test the post-exposure potential of hydroxychloroquine against COVID-19.

Our coalition of public health experts, doctors, and scientists worldwide want to draw attention to the need for high-quality evaluation protocols of the potential beneficial effect of hydroxychloroquine (HCQ) as a post-exposure drug for exposed people. In the absence of an approved, recognized effective pre or post-exposure prophylactic drug or vaccine for COVID-19, nor of any approved and validated therapeutic drug, coupled with social and political pressure raised by publicity both regarding the potential beneficial effect of hydroxychloroquine (HCQ) as well as potential risks from HCQ, we urge the immediate proper clinical trials. Specifically, we mean using HCQ for post-exposure of people with close contact with patients with positive COVID19 rtPCR, including home and medical caregivers. We have reviewed the mechanisms of antiviral effect of HCQ, the risk-benefit ratio taking into consideration the PK/PD of HCQ and the thresholds of efficacy. We have studied its use as an antimalarial, an antiviral, and an immunomodulating drug and concluded that the use of HCQ at doses matching that of the standard treatment of Systemic Lupus erythematous, which has proven safety and efficacy in terms of HCQ blood and tissue concentration adapted to bodyweight (2,3), at 6 mg/kg/day 1 (loading dose) followed by 5 mg/kg/ day, with a maximum limit of 600 mg/day in all cases should swiftly be clinically evaluated as a post-exposure drug for exposed people.

Insecticide-Treated Mosquito Nets.

The coherent and coordinated strategy put in place since the beginning of the century to fight malaria has led to a significant reduction in the global burden of the disease. Of the various elements composing this strategy, it appears that vector control and, in the first place, the massive distribution of LLIN (long-lasting impregnated nets) have contributed significantly to this success. The idea, a priori trivial, of impregnating nets with insecticide to make them more effective, emerged in the late 1980s in Benin. Since then, thanks to the ongoing support of WHO and collaboration with industry, there has been a paradigm shift that puts LLIN at the forefront of malaria control. This chapter summarizes, in chronological order, the different steps that led to the use of LLIN, starting with simple mosquito nets. It provides a brief overview of the studies that have shown their effectiveness both from the entomological and the morbidity and mortality point of view. Finally, it presents the limitations and challenges that will need to be overcome if LLIN are to remain an effective tool in the fight against malaria.

Malaria overdiagnosis and subsequent overconsumption of antimalarial drugs in Angola: Consequences and effects on human health.

Microscopic blood smear examinations done in health centers of Angola demonstrated a large overdiagnosis of malaria cases with an average rate of errors as high as 85%. Overall 83% of patients who received Coartem® had an inappropriate treatment. Overestimated malaria diagnosis was noticed even when specific symptoms were part of the clinical observation, antimalarial treatments being subsequently given. Then, malaria overdiagnosis has three main consequences, (i) the lack of data reliability is of great concern, impeding epidemiological records and evaluation of the actual influence of operations as scheduled by the National Malaria Control Programme; (ii) the large misuse of antimalarial drug can increase the selective pressure for resistant strain and can make a false consideration of drug resistant P. falciparum crisis; and (iii) the need of strengthening national health centers in term of human, with training in microscopy, and equipment resources to improve malaria diagnosis with a large scale use of rapid diagnostic tests associated with thick blood smears, backed up by a "quality control" developed by the national health authorities. Monitoring of malaria cases was done in three Angolan health centers of Alto Liro (Lobito town) and neighbor villages of Cambambi and Asseque (Benguéla Province) to evaluate the real burden of malaria. Carriers of Plasmodium among patients of newly-borne to 14 years old, with or without fever, were analyzed and compared to presumptive malaria cases diagnosed in these health centers. Presumptive malaria cases were diagnosed six times more than the positive thick blood smears done on the same children. In Alto Liro health center, the percentage of diagnosis error reached 98%, while in Cambambi and Asseque it was of 79% and 78% respectively. The percentage of confirmed malaria cases was significantly higher during the dry (20.2%) than the rainy (13.2%) season. These observations in three peripheral health centers confirmed what has already been noticed in other malaria endemic regions, and highlight the need for an accurate evaluation of the Malaria control programme implemented in Angola.

Spatio-temporal variations of Anopheles coluzzii and An. gambiae and their Plasmodium infectivity rates in Lobito, Angola.

From 2003 to 2007, entomological surveys were conducted in Lobito town (Benguela Province, Angola) to determine which Anopheles species were present and to identify the vectors responsible for malaria transmission in areas where workers of the Sonamet Company live. Two types of surveys were conducted: (1) time and space surveys in the low and upper parts of Lobito during the rainy and dry periods; (2) a two-year longitudinal study in Sonamet workers' houses provided with long-lasting insecticide-treated nets (LLIN), "PermaNet," along with the neighboring community. Both species, An. coluzzii (M molecular form) and An. gambiae (S molecular form), were collected. Anopheles coluzzii was predominant during the dry season in the low part of Lobito where larvae develop in natural ponds and temporary pools. However, during the rainy season, An. gambiae was found in higher proportions in the upper part of the town where larvae were collected in domestic water tanks built near houses. Anopheles melas and An. listeri were captured in higher numbers during the dry season and in the low part of Lobito where larvae develop in stagnant brackish water pools. The infectivity rates of An. gambiae s.l. varied from 0.90% to 3.41%.

Pilot study on the combination of an organophosphate-based insecticide paint and pyrethroid-treated long lasting nets against pyrethroid resistant malaria vectors in Burkina Faso.

A pilot study to test the efficacy of combining an organophosphate-based insecticide paint and pyrethroid-treated Long Lasting Insecticide Treated Nets (LLINs) against pyrethroid-resistant malaria vector mosquitoes was performed in a real village setting in Burkina Faso. Paint Inesfly 5A IGR™, comprised of two organophosphates (OPs) and an Insect Growth Regulator (IGR), was tested in combination with pyrethroid-treated LLINs. Efficacy was assessed in terms of mortality for 12 months using Early Morning Collections of malaria vectors and 30-minute WHO bioassays. Resistance to pyrethroids and OPs was assessed by detecting the frequency of L1014F and L1014S kdr mutations and Ace-1(R)G119S mutation, respectively. Blood meal origin was identified using a direct enzyme-linked immunosorbent assay (ELISA). The combination of Inesfly 5A IGR™ and LLINs was effective in killing 99.9-100% of malaria vector populations for 6 months regardless of the dose and volume treated. After 12 months, mortality rates decreased to 69.5-82.2%. The highest mortality rates observed in houses treated with 2 layers of insecticide paint and a larger volume. WHO bioassays supported these results: mortalities were 98.8-100% for 6 months and decreased after 12 months to 81.7-97.0%. Mortality rates in control houses with LLINs were low. Collected malaria vectors consisted exclusively of Anopheles coluzzii and were resistant to pyrethroids, with a L1014 kdr mutation frequency ranging from 60 to 98% through the study. About 58% of An. coluzzii collected inside houses had bloodfed on non-human animals. Combining Inesfly 5A IGR™ and LLINs yielded a one year killing efficacy against An. coluzzii highly resistant to pyrethroids but susceptible to OPs that exhibited an anthropo-zoophilic behaviour in the study area. The results obtained in a real setting supported previous work performed in experimental huts and underscore the need to study the impact that this novel strategy may have on clinical malaria and malaria exposure in children in a similar area of high pyrethroid resistance in South-Western Burkina Faso.

Molecular epidemiology of drug-resistant Plasmodium falciparum in Benguela province, Angola.

BACKGROUND: The malaria situation has been worsening in Angola, partly due to armed conflict until the recent past and drug-resistant Plasmodium falciparum. Malaria transmission is heterogeneous within the country, and data on drug-resistant malaria in different parts of the country are incomplete. The aim of the present study was to evaluate resistance to 4-aminoquinolines and antifolate drugs in P. falciparum isolates collected in Benguela province, central Angola, using molecular markers. METHODS: Fingerprick capillary blood was collected from asymptomatic children aged less than 15 years old during a household survey in and around Balombo town in 2010-2011. Samples were screened for P. falciparum by nested PCR. Molecular markers (P. falciparum dihydrofolate reductase [pfdhfr], P. falciparum dihydropteroate synthase [pfdhps], P. falciparum chloroquine resistance transporter [pfcrt], and P. falciparum multidrug-resistance gene 1 [pfmdr1]) were sequenced to determine the key codons associated with drug resistance. RESULTS: A total of 60 blood samples were positive for P. falciparum. Most isolates with successful PCR amplification had mutant pfdhfr alleles, with either double mutant AICNI (69%) or triple mutant AIRNI (21%) haplotypes. A16V, S108T, and I164L substitutions were not found. Many of the isolates were carriers of either SGKAA (60%) or AGKAA (27%) pfdhps haplotype. K540E substitution was absent. There were only two pfcrt haplotypes: wild-type CVMNK (11%) and mutant CVIET (89%). Wild-type pfmdr1 NYSND haplotype was found in 19% of the isolates, whereas single mutant pfmdr1 YYSND and NFSND haplotypes occurred in 48% and 11%, respectively. Double mutant pfmdr1 haplotypes (YFSND and YYSNY) occurred rarely. CONCLUSIONS: The results suggest that the high prevalence of mutant pfcrt CVIET haplotype is in agreement with low clinical efficacy of chloroquine observed in earlier studies and that the double pfdhfr mutant AICNI and single pfdhps mutant SGKAA are currently the predominant haplotypes associated with antifolate resistance in Benguela province. The hallmark of clinical resistance observed in East Africa, i.e. triple pfdhfr mutant haplotype (AIRNI) and double pfdhps mutant haplotype (SGEAA), was absent. These molecular findings need to be further evaluated in parallel with clinical studies, in particular with the efficacy of intermittent preventive treatment using sulphadoxine-pyrimethamine in pregnant women and artesunate-amodiaquine for uncomplicated malaria.

The Anopheles gambiae cE5 salivary protein: a sensitive biomarker to evaluate the efficacy of insecticide-treated nets in malaria vector control.

Evaluation of vector control is crucial for improving malaria containment and, according to World Health Organization, new complementary indicators would be very valuable. In this study the IgG response to the Anopheles-specific cE5 salivary protein was tested as a tool to evaluate the efficacy of insecticide-treated nets in reducing human exposure to malaria vectors. Sera collected during a longitudinal study carried out in Angola, and including entomological and parasitological data, were used to assess the IgG response to the Anopheles gambiae cE5 in both children and adults, before and after the application of insecticide-treated nets. Seasonal fluctuation of specific IgG antibody levels according to exposure was only found in children (up to ≈ 14 years old) whose anti-cE5 IgG response dropped after bed nets installation. These results were fully consistent with previous findings obtained with the same set of sera and indicating a substantial reduction of human-vector contact shortly after nets implementation. Overall, children IgG response to the cE5 protein appeared a very sensitive biomarker, which allowed for the detection of even weak exposure to Anopheles bites, indicating it may represent a reliable additional tool to evaluate the efficacy of vector control interventions.

Proposed use of spatial mortality assessments as part of the pesticide evaluation scheme for vector control.

BACKGROUND: The WHO Pesticide Evaluation Scheme to evaluate the efficacy of insecticides does not include the testing of a lethal effect at a distance. A tool was developed to evaluate the spatial mortality of an insecticide product against adult mosquitoes at a distance under laboratory and field conditions. Operational implications are discussed. METHODS: Insecticide paint, Inesfly 5A IGR™, containing two organophosphates (OPs): chlorpyrifos and diazinon, and one insect growth regulator (IGR): pyriproxyfen, was the product tested. Laboratory tests were performed using "distance boxes" with surfaces treated with one layer of control or insecticide paint at a dose of 1 kg/6 sq m. Field tests were conducted up to 12 months in six experimental huts randomly allocated to control or one or two layers of insecticide paint at 1 kg/6 sq m. All distance tests were performed using reference-susceptible strains of Anopheles gambiae and Culex quinquefasciatus left overnight at a distance of 1 m from control or treated surfaces. RESULTS: After an overnight exposition at distances of 1 m, field and laboratory evaluations at 0 months after treatment (T0) yielded 100% mortality rates on surfaces treated with one layer at 1 kg/6 sq m against susceptible strains of An. gambiae and Cx. quinquefasciatus. Testing for long-term efficacy in the field gave mortality rates of 96-100% after an overnight exposition at a distance of 1 m for up to 12 months in huts where a larger volume was treated (walls and ceilings) with one or two layers of insecticide paint. CONCLUSION: A comprehensive evaluation of the full profile of insecticide products, both upon contact and spatially, may help rationalize vector control efforts more efficiently. Treating a large enough volume may extend a product's mortality efficacy in the long-term, which contact tests would fail to assess. It is hereby proposed to explore the development of cost effective methods to assess spatial mortality and to include them as one additional measurement of insecticide efficacy against mosquitoes and other arthropod vectors in WHOPES Phase I and Phase II studies.

Inland valley rice production systems and malaria infection and disease in the forest region of western Côte d'Ivoire.

BACKGROUND: This study aimed to determine the epidemiological impact of rice cultivation in inland valleys on malaria in the forest region of western Côte d'Ivoire. The importance of malaria was compared in terms of prevalence and parasite density of infections and also in terms of clinical malaria incidence between three agro-ecosystems: (i) uncultivated inland valleys, (R0), (ii) inland valleys with one annual rice cultivation in the rainy season, (R1) and (iii) developed inland valleys with two annual rice cultivation cycles, (R2). METHODS: Between May 1998 and March 1999, seven villages of each agro-ecosystem (R0, R1 and R2) were randomly selected among villages pooled by farming system. In these 21 villages, a total of 1,900 people of all age groups were randomly selected and clinically monitored during one year. Clinical and parasitological information was obtained by active case detection of malaria episodes carried out during eight periods of five consecutive days scheduled at six weekly intervals and by cross-sectional surveys. RESULTS: Plasmodium falciparum was the principal parasite observed in the three agro-ecosystems. A level of holoendemicity of malaria was observed in the three agro-ecosystems with more than 75% of children less than 12 months old infected. Geometric mean parasite density in asymptomatic persons varied between 180 and 206 P. falciparum asexual forms per µL of blood and was associated with season and with age, but not with farming system. The mean annual malaria incidence rate reached 0.7 (95% IC 0.5-0.9) malaria episodes per person in R0, 0.7 (95% IC 0.6-0.9) in R1 and 0.6 (95% IC 0.5-0.7) in R2. The burden of malaria was the highest among children under two years of age, with at least four attacks by person-year. Then malaria incidence decreased by half in the two to four-year age group. From the age of five years, the incidence was lower than one attack by person-year. Malaria incidence varied with season with more cases in the rainy season than in the dry season but not with farming system. CONCLUSION: In the forest area of western Côte d'Ivoire, inland valley rice cultivation was not significantly associated with malaria burden.

Human antibody response to Anopheles saliva for comparing the efficacy of three malaria vector control methods in Balombo, Angola.

Human antibody (Ab) response to Anopheles whole saliva, used as biomarker of Anopheles exposure, was investigated over a period of two years (2008-2009), in children between 2 to 9 years old, before and after the introduction of three different malaria vector control methods; deltamethrin treated long lasting impregnated nets (LLIN) and insecticide treated plastic sheeting (ITPS)--Zero Fly®) (ITPS-ZF), deltamethrin impregnated Durable (Wall) Lining (ITPS-DL--Zerovector®) alone, and indoor residual spraying (IRS) with lambdacyhalothrin alone. These different vector control methods resulted in considerable decreases in all three entomological (82.4%), parasitological (54.8%) and immunological criteria analyzed. The highest reductions in the number of Anopheles collected and number of positive blood smears, respectively 82.1% and 58.3%, were found in Capango and Canjala where LLIN and ITPS-ZF were implemented. The immunological data based on the level of anti-saliva IgG Ab in children of all villages dropped significantly from 2008 to 2009, except in Chissequele. These results indicated that these three vector control methods significantly reduced malaria infections amongst the children studied and IRS significantly reduced the human-Anopheles contact. The number of Anopheles, positive blood smears, and the levels of anti-saliva IgG Ab were most reduced when LLIN and ITPS-ZF were used in combination, compared to the use of one vector control method alone, either ITPS-DL or IRS. Therefore, as a combination of two vector control methods is significantly more effective than one control method only, this control strategy should be further developed at a more global scale.

Insecticide-treated plastic sheeting for emergency malaria prevention and shelter among displaced populations: an observational cohort study in a refugee setting in Sierra Leone.

A double-blind phase III malaria prevention trial was conducted in two refugee camps using pre-manufactured insecticide-treated plastic sheeting (ITPS) or untreated polyethylene sheeting (UPS) randomly deployed to defined sectors of each camp. In Largo camp the ITPS or UPS was attached to inner walls and ceilings of shelters, whereas in Tobanda the ITPS or UPS was used to line only the ceiling and roof. In Largo the Plasmodium falciparum incidence rate in children up to 3 years of age who were cleared of parasites and monitored for 8 months was 163/100 person-years under UPS and 63 under ITPS (adjusted odds ratio [AOR] = 0.40, 95% confidence interval [CI] = 0.33-0.47). In Tobanda incidence was 157/100 person-years under UPS and 134 under ITPS (AOR = 0.85, 95% CI = 0.75-0.95). Protective efficacy was 61% under fully lined ITPS and 15% under roof lined ITPS. Anemia rates improved under ITPS in both camps. This novel tool proved to be a convenient, safe, and long-lasting method of malaria control when used as a full shelter lining in an emergency setting.

Multiple independent introductions of Plasmodium falciparum in South America.

The origin of Plasmodium falciparum in South America is controversial. Some studies suggest a recent introduction during the European colonizations and the transatlantic slave trade. Other evidence--archeological and genetic--suggests a much older origin. We collected and analyzed P. falciparum isolates from different regions of the world, encompassing the distribution range of the parasite, including populations from sub-Saharan Africa, the Middle East, Southeast Asia, and South America. Analyses of microsatellite and SNP polymorphisms show that the populations of P. falciparum in South America are subdivided in two main genetic clusters (northern and southern). Phylogenetic analyses, as well as Approximate Bayesian Computation methods suggest independent introductions of the two clusters from African sources. Our estimates of divergence time between the South American populations and their likely sources favor a likely introduction from Africa during the transatlantic slave trade.

Human antibody responses to the Anopheles salivary gSG6-P1 peptide: a novel tool for evaluating the efficacy of ITNs in malaria vector control.

To optimize malaria control, WHO has prioritised the need for new indicators to evaluate the efficacy of malaria vector control strategies. The gSG6-P1 peptide from gSG6 protein of Anopheles gambiae salivary glands was previously designed as a specific salivary sequence of malaria vector species. It was shown that the quantification of human antibody (Ab) responses to Anopheles salivary proteins in general and especially to the gSG6-P1 peptide was a pertinent biomarker of human exposure to Anopheles. The present objective was to validate this indicator in the evaluation of the efficacy of Insecticide Treated Nets (ITNs). A longitudinal evaluation, including parasitological, entomological and immunological assessments, was conducted on children and adults from a malaria-endemic area before and after the introduction of ITNs. Significant decrease of anti-gSG6-P1 IgG response was observed just after the efficient ITNs use. Interestingly, specific IgG Ab level was especially pertinent to evaluate a short-time period of ITNs efficacy and at individual level. However, specific IgG rose back up within four months as correct ITN use waned. IgG responses to one salivary peptide could constitute a reliable biomarker for the evaluation of ITN efficacy, at short- and long-term use, and provide a valuable tool in malaria vector control based on a real measurement of human-vector contact.

Efficacy of an insecticide paint against insecticide-susceptible and resistant mosquitoes - part 1: laboratory evaluation.

BACKGROUND: The main malaria vector Anopheles gambiae and the urban pest nuisance Culex quinquefasciatus are increasingly resistant to pyrethroids in many African countries. There is a need for new products and strategies. Insecticide paint Inesfly 5A IGR™, containing two organophosphates (OPs), chlorpyrifos and diazinon, and insect growth regulator (IGR), pyriproxyfen, was tested under laboratory conditions for 12 months following WHOPES Phase I procedures. METHODS: Mosquitoes used were laboratory strains of Cx. quinquefasciatus susceptible and resistant to OPs. The paint was applied at two different doses (1 kg/6 m2 and 1 kg/12 m2) on different commonly used surfaces: porous (cement and stucco) and non-porous (softwood and hard plastic). Insecticide efficacy was studied in terms of delayed mortality using 30-minute WHO bioassay cones. IGR efficacy on fecundity, fertility and larval development was studied on OP-resistant females exposed for 30 minutes to cement treated and control surfaces. RESULTS: After treatment, delayed mortality was high (87-100%) even against OP-resistant females on all surfaces except cement treated at 1 kg/12 m2. Remarkably, one year after treatment delayed mortality was 93-100% against OP-resistant females on non-porous surfaces at both doses. On cement, death rates were low 12 months after treatment regardless of the dose and the resistance status. Fecundity, fertility and adult emergence were reduced after treatment even at the lower dose (p < 10(-3)). A reduction in fecundity was still observed nine months after treatment at both doses (p < 10(-3)) and adult emergence was reduced at the higher dose (p < 10(-3)). CONCLUSIONS: High mortality rates were observed against laboratory strains of the pest mosquito Cx. quinquefasciatus susceptible and resistant to insecticides. Long-term killing remained equally important on non-porous surfaces regardless the resistance status for over 12 months. The paint's effect on fecundity, fertility and adult emergence may continue to provide an additional angle of attack in reducing overall population densities when the lethal effect of OPs diminishes over time. Some options on how to deal with porous materials are given. Implications in vector control are discussed.

Efficacy of an insecticide paint against malaria vectors and nuisance in West Africa--part 2: field evaluation.

BACKGROUND: Widespread resistance of the main malaria vector Anopheles gambiae to pyrethroids reported in many African countries and operational drawbacks to current IRS methods suggest the convenience of exploring new products and approaches for vector control. Insecticide paint Inesfly 5A IGR™, containing two organophosphates (OPs), chlorpyrifos and diazinon, and one insect growth regulator (IGR), pyriproxyfen, was tested in Benin, West Africa, for 12 months. METHODS: Field trials were conducted in six experimental huts that were randomly allocated to one or two layers of insecticide at 1 Kg/6 m2 or control. Evaluations included: (i) early mosquito collection, (ii) mosquito release experiments, (iii) residual efficacy tests and (iv) distance tests. Early mosquito collections were performed on local populations of pyrethroid-resistant An. gambiae and Culex quinquefasciatus. As per WHOPES phase II procedures, four entomological criteria were evaluated: deterrence, excito-repellence, blood-feeding inhibition and mortality. Mosquito release experiments were done using local malaria-free An. gambiae females reared at the CREC insectarium. Residual efficacy tests and distance tests were performed using reference susceptible strains of An. gambiae and Cx. quinquefasciatus. RESULTS: Six months after treatment, mortality rates were still 90-100% against pyrethroid-resistant mosquito populations in experimental huts. At nine months, mortality rates in huts treated with two layers was still about 90-93% against An. gambiae and 55% against Cx. quinquefasciatus. Malaria-free local mosquito release experiments yielded a 90% blood-feeding inhibition in the absence of a physical barrier. A long-term residual efficacy of 12 months was observed by WHO-bioassays in huts treated with two layers (60-80%). Mortality after an overnight exposition at distances of 1 meter was 96-100% for up to 12 months. CONCLUSION: The encouraging results obtained on the insecticide paint Inesfly 5A IGR™ in terms of mortality, be it in direct contact or at a distance, and its new operational approach could constitute an additional option in malaria control efforts in areas of pyrethroid resistance. Phase III studies will be performed to assess the product's epidemiological impact and sociological acceptance.

Human antibody response to Anopheles gambiae saliva: an immuno-epidemiological biomarker to evaluate the efficacy of insecticide-treated nets in malaria vector control.

For the fight against malaria, the World Health Organization (WHO) has emphasized the need for indicators to evaluate the efficacy of vector-control strategies. This study investigates a potential immunological marker, based on human antibody responses to Anopheles saliva, as a new indicator to evaluate the efficacy of insecticide-treated nets (ITNs). Parasitological, entomological, and immunological assessments were carried out in children and adults from a malaria-endemic region of Angola before and after the introduction of ITNs. Immunoglobulin G (IgG) levels to An. gambiae saliva were positively associated with the intensity of An. gambiae exposure and malaria infection. A significant decrease in the anti-saliva IgG response was observed after the introduction of ITNs, and this was associated with a drop in parasite load. This study represents the first stage in the development of a new indicator to evaluate the efficacy of malaria vector-control strategies, which could apply in other arthropod vector-borne diseases.

Evaluation of synthetic repellents on mosquito nets in experimental huts against insecticide-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes.

Owing to the development of pyrethroid resistance in Anopheles gambiae, there is a need to develop chemical alternatives for use on mosquito nets. Synthetic insect repellents are widely used for personal protection as skin or clothing applications. The efficacy of repellent-treated nets (RTN) was evaluated in experimental huts in Côte d'Ivoire against pyrethroid-resistant populations of An. gambiae and Culex quinquefasciatus. The repellents tested were DEET (N,N-diethyl-3-methylbenzamide) at 7.9 g/m2 and two formulations of ethyl butylacetylaminopropionate (IR3535) at 7.6 g/m2 and 7.3 g/m2. Over 45 nights there was a 74-82% reduction in the number of An. gambiae entering the huts containing RTNs but no significant reduction in entry of C. quinquefasciatus. There was a 63-64% reduction in the proportion of An. gambiae blood feeding but no reduction in the proportion of C. quinquefasciatus blood feeding in huts with RTNs. An unexpected result was the 69-76% mortality of An. gambiae and 51-61% mortality of C. quinquefasciatus in huts containing RTNs. Treated filter paper bioassays in WHO test kits confirmed that confined contact with DEET induces mortality. The DEET-based product provided better and longer protection; tunnel test bioassays confirmed that residual activity lasted for up to 6 weeks. Application of repellents to nets warrants further investigation and development.