Seroepidemiology of toxoplasmosis in pregnant women and detection of infection acquired during pregnancy in Cotonou, Benin.

Assessing the prevalence of toxoplasmosis in pregnant women and the associated risk factors is the first step in defining policy for the prevention of congenital toxoplasmosis in a given population. An epidemiological study was conducted during prenatal consultations at the CHU-MEL of Cotonou (Benin) between September 2018 and April 2021 and recruited 549 pregnant women to determine the seroprevalence and potential factors associated with Toxoplasma gondii infection. Toxoplasma gondii IgG/IgM antibodies were detected using an enzyme-linked fluorescence assay (ELFA) technique, an IgG avidity test and an IgG/IgM comparative Western blot to diagnose the maternal toxoplasmosis serological status, the possibility of an infection acquired during pregnancy and congenital infection, respectively. Concomitantly, the participants answered a questionnaire investigating potential risk factors. Toxoplasmosis seroprevalence was estimated at 44.4% (95% CI 40.3-48.6) and the factors significantly associated with T. gondii seropositivity were: age over 30 years, multigravid women and contact with cats. The possibility of an infection acquired during the periconceptional period or the first trimester of pregnancy concerned six women [1.1% (95% CI 0.5-2.0)]. However, due to the low rate of serological controls in seronegative women, a significant proportion of women first tested during the 3rd trimester of pregnancy, and an insufficient sample size, the incidence of primary infection during pregnancy could not be determined. No cases of congenital transmission occurred in the newborns from the suspected cases of primary infection.

Title: Séroépidémiologie de la toxoplasmose chez la femme enceinte et détection de l'infection contractée pendant la grossesse à Cotonou, Bénin. Abstract: L'évaluation de la prévalence de la toxoplasmose chez la femme enceinte et des facteurs de risque associés est la première étape pour définir une politique de prévention de la toxoplasmose congénitale dans une population donnée. Une étude épidémiologique a été menée lors des consultations prénatales au CHU-MEL de Cotonou (Bénin) entre septembre 2018 et avril 2021 et a recruté 549 femmes enceintes pour déterminer la séroprévalence et les facteurs potentiels associés à l'infection à Toxoplasma gondii. Les anticorps IgG / IgM de T. gondii ont été détectés à l'aide d'une technique ELFA, du test d'avidité IgG et du Western blot comparatif IgG / IgM pour diagnostiquer respectivement le statut sérologique de la toxoplasmose maternelle, la possibilité d'une infection acquise pendant la grossesse et l'infection congénitale. Parallèlement, les participants ont répondu à un questionnaire portant sur les facteurs de risque potentiels. La séroprévalence de la toxoplasmose a été estimée à 44,4 % (IC 95 % 40,3­48,6) et les facteurs significativement associés à la séropositivité pour T. gondii étaient l'âge supérieur à 30 ans, la multigravidité et les contacts avec les chats. La possibilité d'une infection acquise pendant la période périconceptionnelle ou le premier trimestre de la grossesse concernait six femmes [1,1 % (IC 95 % 0,5­2,0)]. Cependant, en raison du faible taux de contrôles sérologiques chez les femmes séronégatives, d'une proportion importante de femmes testées pour la première fois au cours du 3ème trimestre de la grossesse et d'une taille d'échantillon insuffisante, l'incidence de la primo-infection pendant la grossesse n'a pas pu être déterminée. Aucun des enfants nés des six femmes suspectes de primo-infection en cours de grossesse n'a présenté d'infection congénitale.

[Influence of white blood cells count on parasite density in Malaria in children aged 6 to 59 months in Benin]. / Influence de la numération leucocytaire sur la densité parasitaire dans le paludisme simple chez les enfants de 6 à 59 mois au Bénin.

Background: For many years, the treatment of malaria was based on clinical presumptive diagnosis, making its differential diagnosis with other causes of hyperthermia difficult. This drug pressure has led to the emergence of Plasmodium strains resistant to the most commonly used antimalarial drugs. This is why in 2004, the health authorities decided to revise the policy of malaria management by adopting a new strategy based on the rational use of artemisininbased combination therapies after the biological confirmation of suspected malaria cases. The biological diagnosis is an essential part of malaria management. The gold standard technique for diagnosis is the thick drop combined with the calculation of parasite density (PD), which is determined on the basis of the number of parasites counted in a microscopic field against a proposed standard number of leukocytes. The number of leukocytes used to calculate the parasite density should ideally be the actual number of leukocytes in the patient per cubic millimetre of blood. However, in the absence of the availability of a blood count at the time of the thick drop, an average number of 8 000 leukocytes/mm3 was used by the World Health Organisation (WHO) to estimate the parasite density. Nonetheless, in Benin the average number of leukocytes adopted by the National Malaria Control Programme (PNLP) is 6 000/mm3. The aim of our study was to determine the impact of the leukocyte count on the calculation of the parasite density in cases of uncomplicated malaria. Method: The study was a cross-sectional study with an analytical aim and took place in 2 hospitals in Benin, the Klouékanmey zone hospital in the south of Benin and the Djougou health centre in the north. It involved a population of 476 children aged between 6 and 59 months who were seen in consultation and in whom the clinical diagnosis of simple Plasmodium falciparum malaria was suspected. Children aged between 6 and 59 months, weighing at least 5 kg, with an axillary temperature ≥ 37.5°C at the time of consultation or a history of fever in the last 24 hours or other symptoms pointing to the diagnosis of malaria were included. Infestation was mono-specific for Plasmodium falciparum. Informed consent was required from the child's parents or guardian. The criteria for non-inclusion in our study were the presence of at least one sign of malaria severity, signs of severe malnutrition or a febrile state related to underlying infectious diseases other than malaria. Thick blood count and haemogram were systematically performed in all included children. Parasite density was calculated according to 3 methods, first using a weighted leukocyte count of 6 000/mm3 recommended by the Benin National Malaria Control Programme (PNLP), then a leukocyte count of 8 000/mm3 recommended by the World Health Organisation and finally the patient's actual leukocyte count obtained from the blood count. It should be noted that these different samples were respectively taken on the day of inclusion in compliance with the conditions of the pre-analytical phase in force in our medical biology laboratory. Results: At the end of our study, 313 children, i.e. 65.76% of our study population had a positive white blood cell count with a positivity rate of 62.14% in Djougou, i.e. 174 children, and 70.9% in Klouékanmey, i.e. 139 children. The average leukocyte count in these children was 11,580/mm3. Among them, 205 children had an abnormal white blood cell count, i.e. 17 cases of leukopenia (5.43%) and 188 cases of hyperleukocytosis (60.06%). Using successively the average number of 6 000 leukocytes/mm3 proposed by the Benin PNLP and that of 8 000 leukocytes/mm3 proposed by the WHO, the average parasite densities were respectively 47,943 and 63,936 trophozoïtes/µl against 92,290 trophozoïtes/µl when the real number of leukocytes of the patients was used for the calculation of the PD. By using an average of 6 000 leukocytes/mm3 for PD calculation, 60% of the calculated PDs were underestimated and 6% were overestimated. Using an average of 8 000 leukocytes/mm3 resulted in 49% of PD being underestimated and 15% being overestimated. The difference between the three calculation methods was considered statistically significant (p value <0.05). Conclusion: The use of 6 000 or 8 000 coefficients for the estimation of parasitaemia could lead to a significant underestimation of the parasite load.

Implications of insecticide resistance for malaria vector control with long-lasting insecticidal nets: evidence from health facility data from Benin.

BACKGROUND: Insecticide-based interventions have averted more than 500 million malaria cases since 2000, but insecticide resistance in mosquitoes could bring about a rebound in disease and mortality. This study investigated whether insecticide resistance was associated with increased incidence of clinical malaria. METHODS: In an area of southern Benin with insecticide resistance and high use of insecticide-treated nets (ITNs), malaria morbidity and insecticide resistance were measured simultaneously in 30 clusters (villages or collections of villages) multiple times over the course of 2 years. Insecticide resistance frequencies were measured using the standard World Health Organization bioassay test. Malaria morbidity was measured by cases recorded at health facilities both in the whole population using routinely collected data and in a passively followed cohort of children under 5 years old. RESULTS: There was no evidence that incidence of malaria from routinely collected data was higher in clusters with resistance frequencies above the median, either in children aged under 5 (RR = 1.27 (95% CI 0.81-2.00) p = 0.276) or in individuals aged 5 or over (RR = 1.74 (95% CI 0.91-3.34) p = 0.093). There was also no evidence that incidence was higher in clusters with resistance frequencies above the median in the passively followed cohort (RR = 1.11 (0.52-2.35) p = 0.777). CONCLUSIONS: This study found no association between frequency of resistance and incidence of clinical malaria in an area where ITNs are the principal form of vector control. This may be because, as other studies have shown, ITNs continue to offer some protection from malaria even in the presence of insecticide resistance. Irrespective of resistance, nets provide only partial protection so the development of improved or supplementary vector control tools is required to reduce Africa's unacceptably high malaria burden.

Results of the first mapping of soil-transmitted helminths in Benin: Evidence of countrywide hookworm predominance.

BACKGROUND: National mapping of soil-transmitted helminth infections (STH) was conducted for the first time in all of the 77 districts of Benin (West Africa) from 2013 to 2015. This mapping aimed to provide basic epidemiological data essential for the implementation of the national strategy against the neglected tropical diseases (NTDs) in the context of achieving the WHO target of controlling these infections by 2020. METHODS: In each district, 5 schools were purposively selected in 5 villages and 50 school-children (25 girls and 25 boys) from ages 8 to 14 years were randomly enrolled in each school. In total, 19,250 stool samples of school children (9,625 girls and 9,625 boys) from 385 schools were examined by Kato-Katz technique. RESULTS: The three major species of STH (hookworm, Ascaris lumbricoides and Trichuris trichiura) were observed with intra- and inter-specific variations in the prevalence and the intensity of these parasites. Hookworm infection was present in all of the surveyed districts with an average prevalence of 17.14% (95% CI 16.6%-17.6%). Among the infected schoolchildren, at national level, 90.82%, 6.73% and 2.45% of infections were of light, moderate and heavy parasite intensities respectively. A. lumbricoides infection, with a national average prevalence of 5.35% (95% CI 5.00%-5.60%),was the second most prevalent STH, and 84.37%, 14.27% and 1.36% of the infections were of light, moderate and heavy parasite intensities, respectively. T. trichiura had a national average prevalence of 1.15% (95% CI 0.90%-1.20%) and 80.45%, 13.18% and 6.36% infections were of light, moderate and heavy parasite intensities, respectively. The national cumulative prevalence of the three STH infections was 22.74% (95% CI 22.15%-23.33%), with58.44% (45/77) of the districts requiring mass treatment according to WHO recommendations. In all of the surveyed districts, multiple infections by STH species were common, and boys seemed more at risk of hookworm and Ascaris infections. CONCLUSIONS: This first national mapping provided an overview of the epidemiological pattern of STH infections and was essential for the implementation of a control strategy with an effective preventive chemotherapy treatment (PCT). Results show that while preventive chemotherapy is not indicated for children in 32/77 districts, 43 require annual deworming and two require twice yearly deworming. If no environmental change occurs, and no mass treatment is delivered, prevalence is likely to remain stable for many years owing to poor hygiene and sanitation.

Insecticide-treated nets provide protection against malaria to children in an area of insecticide resistance in Southern Benin.

BACKGROUND: Malaria control is heavily reliant on insecticides, especially pyrethroids. Resistance of mosquitoes to insecticides may threaten the effectiveness of insecticide-based vector control and lead to a resurgence of malaria in Africa. METHODS: In 21 villages in Southern Benin with high levels of insecticide resistance, the resistance status of local vectors was measured at the same time as the prevalence of malaria infection in resident children. RESULTS: Children who used LLINs had lower levels of malaria infection [odds ratio = 0.76 (95% CI 0.59, 0.98, p = 0.033)]. There was no evidence that the effectiveness of nets was different in high and low resistance locations (p = 0.513). There was no association between village level resistance and village level malaria prevalence (p = 0.999). CONCLUSIONS: LLINs continue to offer individual protection against malaria infection in an area of high resistance. Insecticide resistance is not a reason to stop efforts to increase coverage of LLINs in Africa.

Prevalence of Trachoma in Northern Benin: Results from 11 Population-Based Prevalence Surveys Covering 26 Districts.

AIMS: We sought to evaluate trachoma prevalence in all suspected-endemic areas of Benin. METHODS: We conducted population-based surveys covering 26 districts grouped into 11 evaluation units (EUs), using a two-stage, systematic and random, cluster sampling design powered at EU level. In each EU, 23 villages were systematically selected with population proportional to size; 30 households were selected from each village using compact segment sampling. In selected households, we examined all consenting residents aged one year or above for trichiasis, trachomatous inflammation - follicular (TF), and trachomatous inflammation - intense. We calculated the EU-level backlog of trichiasis and delineated the ophthalmic workforce in each EU using local interviews and telephone surveys. RESULTS: At EU-level, the TF prevalence in 1-9-year-olds ranged from 1.9 to 24.0%, with four EUs (incorporating eight districts) demonstrating prevalences ≥5%. The prevalence of trichiasis in adults aged 15+ years ranged from 0.1 to 1.9%. In nine EUs (incorporating 19 districts), the trichiasis prevalence in adults was ≥0.2%. An estimated 11,457 people have trichiasis in an area served by eight ophthalmic clinical officers. CONCLUSION: In northern Benin, over 8000 people need surgery or other interventions for trichiasis to reach the trichiasis elimination threshold prevalence in each EU, and just over one million people need a combination of antibiotics, facial cleanliness and environmental improvement for the purposes of trachoma's elimination as a public health problem. The current distribution of ophthalmic clinical officers does not match surgical needs.

Therapeutic efficacy of artemether-lumefantrine for the treatment of uncomplicated falciparum malaria in northwest Benin.

BACKGROUND: Artemether/lumefantrine (Coartem(®)) has been used as a treatment for uncomplicated Plasmodium falciparum infection since 2004 in Benin. This open-label, non-randomized study evaluated efficacy of artemether-lumefantrine (AL) in treatment of uncomplicated falciparum malaria in children aged 6-59 months in two malaria transmission sites in northwest Benin. METHODS: A 42-day therapeutic efficacy study was conducted between August and November 2014, in accordance with 2009 WHO guidelines. One-hundred and twenty-three children, aged 6 months to 5 years, with uncomplicated falciparum malaria were recruited into the study. The primary endpoint was parasitological cure on day 28 and day 42 while the secondary endpoints included: parasite and fever clearance, improvement in haemoglobin levels. Outcomes were classified as early treatment failure (ETF), late clinical failure, late parasitological failure, and adequate clinical and parasitological response (ACPR). RESULTS: Before PCR correction, ACPR rates were 87% (95 % CI 76.0-94.7) and 75.6%, respectively (95% CI 67.0-82.9) on day 28 and day 42. In each study site, ACPR rates were 78.3% in Djougou and 73% in Cobly on day 42. There was no ETF and after PCR correction ACPR was 100% in study population. All treatment failures were shown to be due to new infections. Fever was significantly cleared in 24 h and approximately 90% of parasites where cleared on day 1 and almost all parasites were cleared on day 2. Haemoglobin concentration showed a slight increase with parasitic clearance. CONCLUSION: AL remains an efficacious drug for the treatment of uncomplicated falciparum malaria in Benin, although higher rates of re-infection remain a concern. Surveillance needs to be continued to detect future changes in parasite sensitivity to artemisinin-based combination therapy.

Prevalence of nosocomial infections and anti-infective therapy in Benin: results of the first nationwide survey in 2012.

BACKGROUND: Data on nosocomial infections in hospitals in low-income countries are scarce and often inconsistent. The objectives of this study were to estimate the prevalence of nosocomial infections and antimicrobial drug use in Benin hospitals. METHODS: All hospitals were invited to participate in the first national point prevalence study conducted between 10-26 October 2012 using the protocol developed by the "Hospitals in Europe Link for Infection Control through Surveillance" (HELICS) project. Infection prevalence rates and the proportion of infected patients and exposure to antimicrobials were assessed. RESULTS: Overall, 87% (39/45) of hospitals participated. Of 3130 inpatients surveyed, 972 nosocomial infections were identified among 597 patients, representing an overall prevalence of infected patients of 19.1%. The most frequent infections were related to the urinary tract (48.2%), vascular catheter use (34.7%), and surgical site (24.7%). 64.6% of patients surveyed were treated with antibiotics, including a significant proportion (30%) of non-infected patients and a high proportion of self-medication (40.8%). Resistance of leading nosocomial pathogens to antimicrobials included methicillin-resistance (52.5%) among Staphylococcus aureus, vancomycin resistance among enterococci (67.5%), cefotaxime resistance among Escherichia coli (67.6%), and ceftazidime resistance among Acinetobacter baumannii (100%) and Pseudomonas aeruginosa (68.2%). CONCLUSIONS: Benin has high nosocomial infection rates and calls for the implementation of new national infection control policies. Patient safety education and training of all individuals involved in healthcare delivery will be critical to highlight awareness of the burden of disease. The high use of antimicrobials needs to be addressed, particularly their indiscriminate use in non-infected patients.

Impact of long-lasting, insecticidal nets on anaemia and prevalence of Plasmodium falciparum among children under five years in areas with highly resistant malaria vectors.

BACKGROUND: The widespread use of insecticide-treated nets (LLINs) leads to the development of vector resistance to insecticide. This resistance can reduce the effectiveness of LLIN-based interventions and perhaps reverse progress in reducing malaria morbidity. To prevent such difficulty, it is important to know the real impact of resistance in the effectiveness of mosquito nets. Therefore, an assessment of LLIN efficacy was conducted in malaria prevention among children in high and low resistance areas. METHODS: The study was conducted in four rural districts and included 32 villages categorized as low or high resistance areas in Plateau Department, south-western Benin. Larvae collection was conducted to measure vector susceptibility to deltamethrin and knockdown resistance (kdr) frequency. In each resistance area, around 500 children were selected to measure the prevalence of malaria infection as well as the prevalence of anaemia associated with the use of LLINs. RESULTS: Observed mortalities of Anopheles gambiae s.s population exposed to deltamethrin ranged from 19 to 96%. Knockdown resistance frequency was between 38 and 84%. The prevalence of malaria infection in children under five years was 22.4% (19.9-25.1). This prevalence was 17.3% (14.2-20.9) in areas of high resistance and 27.1% (23.5-31.1) in areas of low resistance (p=0.04). Eight on ten children that were aged six - 30 months against seven on ten of those aged 31-59 months were anaemic. The anaemia observed in the six to 30-month old children was significantly higher than in the 31-59 month old children (p=0.00) but no difference associated with resistance areas was observed (p=0.35). The net use rate was 71%. The risk of having malaria was significantly reduced (p<0.05) with LLIN use in both low and high resistance areas. The preventive effect of LLINs in high resistance areas was 60% (95% CI: 40-70), and was significantly higher than that observed in low resistance areas (p<0.05). CONCLUSION: The results of this study showed that the resistance of malaria vectors seems to date not have affected the impact of LLINs and the use of LLINs was highly associated with reduced malaria prevalence irrespective of resistance.

Low prevalence of the molecular markers of Plasmodium falciparum resistance to chloroquine and sulphadoxine/pyrimethamine in asymptomatic children in Northern Benin.

BACKGROUND: In Benin, very few studies have been done on the genetics of Plasmodium falciparum and the resistance markers of anti-malarial drugs, while malaria treatment policy changed in 2004. Chloroquine (CQ) and sulphadoxine pyrimethamine (SP) have been removed and replaced by artemisinin-combination therapy (ACT). The objective of this study was to determine the genetic diversity of P. falciparum and the prevalence of P. falciparum molecular markers that are associated with resistance to CQ and SP in northern Benin seven years after the new policy was instituted. METHODS: The study was conducted in northern Benin, a region characterized by a seasonal malaria transmission. Blood samples were collected in 2012 from children presenting with asymptomatic P. falciparum infections. Samples collected in filter paper were genotyped by primary and nested PCR in block 2 of msp-1 and block 3 of msp-2 to analyse the diversity of P. falciparum. The prevalence of critical point mutations in the genes of Pfcrt (codon 76), Pfmdr1 (codon 86), Pfdhfr (codons, 51, 59 and 108) and Pfdhps (codons 437, 540) was examined in parasite isolates by mutation-specific restriction enzyme digestion. RESULTS: Genotyping of 195 isolates from asymptomatic children showed 34 msp-1 and 38 msp-2 genotypes. The multiplicity of infection was 4.51 ± 0.35 for msp-1 and 4.84 ± 0.30 for msp-2. Only the codon 51 of Pfdhfr and codon 437 of Pfdhps showed a high mutation rate: I51: 64.4% (57.3; 71.2); G437: 47.4% (40.2; 54.7), respectively. The prevalence of Pfdhfr triple mutant IRN (I51, R59 and N108) was 1.5% (0.3; 3.9), and Pfdhfr/Pfdhps quadruple mutant IRNG (PfdhfrI51, R59, N108, and PfdhpsG437): 0. 5% (0; 2.5). No mutation was found with codon 540 of Pfdhps. Analysis of mutation according to age (younger or older than ten years) showed similar frequencies in each category without significant difference between the two groups. CONCLUSIONS: This study showed a high diversity of P. falciparum in northern Benin with a very low prevalence of resistance markers to CQ and SP that dramatically contrasted with the pattern observed in southern Benin. No influence of age on genetic diversity of P. falciparum and on distribution of the mutations was observed.

Prevalence of the molecular marker of Plasmodium falciparum resistance to chloroquine and sulphadoxine/pyrimethamine in Benin seven years after the change of malaria treatment policy.

BACKGROUND: In Benin, the National Malaria Control Programme (NMCP) changed the policy of malaria treatment in 2004 following increasing of failure rate of treatment with chloroquine (CQ) and sulphadoxine-pyrimethamine (SP). The objective of this study was to determinate the prevalence of Plasmodium falciparum molecular markers that are associated with resistance to CQ and SP in Benin seven years after the new policy was instituted. METHODS: The study was conducted in southern Benin, a region characterized by a perennial malaria transmission. Blood samples were collected in 2011 from children presenting with symptomatic and asymptomatic P. falciparum infections and living in the same area. The prevalence of critical point mutations in the genes of pfcrt (codon 76), pfmdr1 (codon 86), pfdhfr (codons, 51, 59 and 108) and pfdhps (codons 437, 540) was examined in parasite isolates by mutation-specific restriction enzyme digestion of nested PCR products. RESULTS: A high prevalence of parasites carrying point mutations in all studied targets was found: T76: 93.9% [89.8; 96.7], I51: 96.2% [92.7; 98.4], R59: 93, 9% [89.7; 96.7], N108: 97.6% [94.6; 99.2] and G437: 71.4% [64.8; 77.4]. No mutation was found at codon 540 of the pfdhps gene. The proportion of parasite isolates carrying triple mutation in the pfdhfr gene IRN (I51, R59 andN108) and quadruple mutation on the combination of pfdhfr/pfdhps IRNG (I51, R59, N108 and G437) was 91.5% [86.9; 94.9] and 65.7% [58.9; 72.1], respectively. Analysis of mutation in relation to the clinical status (symptomatic or asymptomatic) and according to age (younger or older than 10 years) showed similar very high frequencies in each category without significant difference between two groups. CONCLUSIONS: These results suggest a persistence level of resistance of P. falciparum to CQ and SP, seven years after the recommendation of the change of malaria treatment policy in Benin. The distribution of mutations studied was neither related to age nor to clinical status.

Malaria infection and disease in an area with pyrethroid-resistant vectors in southern Benin.

BACKGROUND: This study aimed to investigate baseline data on malaria before the evaluation of new vector control strategies in an area of pyrethroid-resistance of vectors. The burden of malaria was estimated in terms of infection (prevalence and parasite density) and of clinical episodes. METHODS: Between December 2007 and December 2008 in the health district of Ouidah-Kpomassè-Tori Bossito (southern Benin), a descriptive epidemiological survey of malaria was conducted. From 28 selected villages, seven were randomized from which a total of 440 children aged 0 to 5 years were randomly selected. Clinical and parasitological information was obtained by active case detection of malaria episodes carried out during eight periods of six consecutive days scheduled at six weekly intervals and by cross-sectional surveys of asymptomatic infection. Entomological information was also collected. The ownership, the use and the correct use of long-lasting insecticide-treated nets (LLINs) were checked over weekly-survey by unannounced visits at home in the late evening. RESULTS: Mean parasite density in asymptomatic children was 586 P. falciparum asexual forms per µL of blood (95%CI 504-680). Pyrogenic parasite cut-off was estimated 2,000 P. falciparum asexual blood forms per µL. The clinical incidence of malaria was 1.5 episodes per child per year (95%CI 1.2-1.9). Parasitological and clinical variables did not vary with season. Anopheles gambiae s.l. was the principal vector closely followed by Anopheles funestus. Entomological inoculation rate was 5.3 (95%CI 1.1-25.9) infective bites per human per year. Frequency of the L1014F kdr (West) allele was around 50%. Annual prevalence rate of Plasmodium falciparum asymptomatic infection was 21.8% (95%CI 19.1-24.4) and increased according to age. Mean rates of ownership and use of LLINs were 92% and 70% respectively. The only correct use of LLINs (63%) conferred 26% individual protection against only infection (OR = 0.74 (95%IC 0.62-0.87), p = 0.005). CONCLUSION: The health district of Ouidah-Kpomassè-Tori Bossito is a mesoendemic area with a moderate level of pyrethroid-resistance of vectors. The used LLINs rate was high and only the correct use of LLINs was found to reduce malaria infection without influencing malaria morbidity.

Dramatically decreased therapeutic efficacy of chloroquine and sulfadoxine-pyrimethamine, but not mefloquine, in southern Benin.

OBJECTIVE: To evaluate the in vivo therapeutic efficacy of chloroquine (CQ), sulfadoxine-pyrimethamine (SP) and mefloquine (MQ) in children presenting with uncomplicated malaria in Benin. METHODS: Drug efficacy was tested according to the WHO in vivo 28-day protocol. For failures that occurred after 7 days of follow-up, paired pre- and post-treatment blood samples were genotyped at msp1 and msp2 loci to distinguish new infections and recrudescent strains. Children enrolled were randomly assigned to a therapeutic group (CQ, n=14; SP, n=42; MQ, n=44). The number of CQ treatment was intentionally restricted after 1 month, as its use was considered to constitute a danger for children. RESULTS: Chloroquine and SP showed very high failure rates (85.7% and 50%, respectively), whereas MQ treatment was successful in 97.5%. The molecular tool allowed to re-evaluate two new infections previously considered as failures. CONCLUSIONS: Chloroquine should no longer be used to treat children presenting with Plasmodium falciparum malaria in Benin.

Screening of pesticide residues in soil and water samples from agricultural settings.

BACKGROUND: The role of agricultural practices in the selection of insecticide resistance in malaria vectors has so far been hypothesized without clear evidence. Many mosquito species, Anopheles gambiae in particular, lay their eggs in breeding sites located around agricultural settings. There is a probability that, as a result of farming activities, insecticide residues may be found in soil and water, where they exercise a selection pressure on the larval stage of various populations of mosquitoes. To confirm this hypothesis, a study was conducted in the Republic of Benin to assess the environmental hazards which can be generated from massive use of pesticides in agricultural settings. METHODS: Lacking an HPLC machine for direct quantification of insecticide residues in samples, this investigation was performed using indirect bioassays focussed on the study of factors inhibiting the normal growth of mosquito larvae in breeding sites. The speed of development was monitored as well as the yield of rearing An. gambiae larvae in breeding sites reconstituted with water and soil samples collected in agricultural areas known to be under pesticide pressure. Two strains of An. gambiae were used in this indirect bioassay: the pyrethroid-susceptible Kisumu strain and the resistant Ladji strain. The key approach in this methodology is based on comparison of the growth of larvae in test and in control breeding sites, the test samples having been collected from two vegetable farms. RESULTS: Results obtained clearly show the presence of inhibiting factors on test samples. A normal growth of larvae was observed in control samples. In breeding sites simulated by using a few grams of soil samples from the two vegetable farms under constant insecticide treatments (test samples), a poor hatching rate of Anopheles eggs coupled with a retarded growth of larvae and a low yield of adult mosquitoes from hatched eggs, was noticed. CONCLUSION: Toxic factors inhibiting the hatching of anopheles eggs and the growth of larvae are probably pesticide residues from agricultural practices. Samples used during this indirect assay have been stored in the laboratory and will be analysed with HPLC techniques to confirm hypothesis of this study and to identify the various end products found in soil and water samples from agricultural settings under pesticide pressure.