Species composition and insecticide resistance in malaria vectors in Ellibou, southern Côte d'Ivoire and first finding of Anopheles arabiensis in Côte d'Ivoire.

BACKGROUND: Knowing the species composition and insecticide resistance status of the target vector population is important to guide malaria vector control. The aim of this study was to characterize the malaria vector population in terms of species composition, insecticide susceptibility status and potential underlying resistance mechanisms in Ellibou, southern Côte d'Ivoire. METHODS: A 1-year longitudinal entomological survey was conducted using light traps and pyrethroid spray catches to sample adult mosquitoes in combination with larval sampling. The susceptibility status of Anopheles gambiae sensu lato (s.l.) to bendiocarb, deltamethrin, DDT and malathion was assessed using the World Health Organization insecticide susceptibility test. Additionally, An. gambiae specimens were screened for knockdown (kdr) and acetylcholineesterase (ace1) target site resistance alleles, and the expression levels of eight metabolic resistance genes, including seven cytochrome P450 monooxygenases (P450s) and one glutathione S-transferase (GST), measured with reverse transcription quantitative real-time polymerase chain reaction (qPCR). RESULTS: Overall, 2383 adult mosquitoes from 12 different taxa were collected with Culex quinquefasciatus and An. gambiae being the predominant taxa. Molecular identification of An. gambiae s.l. revealed the presence of Anopheles arabiensis, Anopheles coluzzii, An. gambiae sensu stricto (s.s.) and Anopheles coluzzii/An. gambiae s.s. hybrids. Anopheles gambiae mosquitoes were resistant to all insecticides except malathion. PCR diagnostics revealed the presence of ace1-G280S and the kdr L995F, L995S and N1570Y target-site mutations. Additionally, several genes were upregulated, including five P450s (i.e., CYP6P3, CYP6M2, CYP9K1, CYP6Z1, CYP6P1) and GSTE2. CONCLUSION: This is the first documented presence of An. arabiensis in Côte d'Ivoire. Its detection - together with a recent finding further north of the country - confirms its existence in the country, which is an early warning sign, as An. arabiensis shows a different biology than the currently documented malaria vectors. Because the local An. gambiae population was still susceptible to malathion, upregulation of P450s, conferring insecticide resistance to pyrethroids, together with the presence of ace1, suggest negative cross-resistance. Therefore, organophosphates could be an alternative insecticide class for indoor residual spraying in the Ellibou area, while additional tools against the outdoor biting An. arabiensis will have to be considered.

Population genetic structure of Schistosoma haematobium and Schistosoma haematobium × Schistosoma bovis hybrids among school-aged children in Côte d'Ivoire.

While population genetics of Schistosoma haematobium have been investigated in West Africa, only scant data are available from Côte d'Ivoire. The purpose of this study was to analyze both genetic variability and genetic structure among S. haematobium populations and to quantify the frequency of S. haematobium × S. bovis hybrids in school-aged children in different parts of Côte d'Ivoire. Urine samples were subjected to a filtration method and examined microscopically for Schistosoma eggs in four sites in the western and southern parts of Côte d'Ivoire. A total of 2692 miracidia were collected individually and stored on Whatman® FTA cards. Of these, 2561 miracidia were successfully genotyped for species and hybrid identification using rapid diagnostic multiplex mitochondrial cox1 PCR and PCR Restriction Fragment Length Polymorphism (PCR-RFLP) analysis of the nuclear ITS2 region. From 2164 miracidia, 1966 (90.9%) were successfully genotyped using at least 10 nuclear microsatellite loci to investigate genetic diversity and population structure. Significant differences were found between sites in all genetic diversity indices and genotypic differentiation was observed between the site in the West and the three sites in the East. Analysis at the infrapopulation level revealed clustering of parasite genotypes within individual children, particularly in Duekoué (West) and Sikensi (East). Of the six possible cox1-ITS2 genetic profiles obtained from miracidia, S. bovis cox1 × S. haematobium ITS2 (42.0%) was the most commonly observed in the populations. We identified only 15 miracidia (0.7%) with an S. bovis cox1 × S. bovis ITS2 genotype. Our study provides new insights into the population genetics of S. haematobium and S. haematobium × S. bovis hybrids in humans in Côte d'Ivoire and we advocate for researching hybrid schistosomes in animals such as rodents and cattle in Côte d'Ivoire.

Title: Structuration génétique des populations de Schistosoma haematobium et des hybrides Schistosoma haematobium × Schistosoma bovis chez les enfants d'âge scolaire en Côte d'Ivoire. Abstract: Alors que la génétique des populations de Schistosoma haematobium a été étudiée en Afrique de l'Ouest, seules quelques données sont disponibles pour la Côte d'Ivoire. Le but de cette étude était d'analyser à la fois la variabilité génétique et la structure génétique des populations de S. haematobium et de quantifier la fréquence des hybrides S. haematobium × S. bovis chez les enfants d'âge scolaire dans différentes régions de la Côte d'Ivoire. Des échantillons d'urine ont été soumis à une méthode de filtration et examinés au microscope pour les œufs de Schistosoma dans quatre sites de l'ouest et du sud de la Côte d'Ivoire. Au total, 2 692 miracidia ont été collectés individuellement et stockés sur des cartes Whatman® FTA. Parmi ceux-ci, 2 561 miracidia ont été génotypés avec succès pour l'identification des espèces et des hybrides à l'aide de la PCR multiplex de diagnostic rapide du cox1 mitochondrial et d'une analyse du polymorphisme de longueur des fragments de restriction de PCR (PCR-RFLP) de la région ITS2 de l'ADN nucléaire. Sur 2 164 miracidia, 1 966 (90,9 %) ont été génotypés avec succès en utilisant au moins 10 loci microsatellites nucléaires pour étudier la diversité génétique et la structure de la population. Des différences significatives ont été trouvées entre les sites dans tous les indices de diversité génétique et une différenciation génotypique a été observée entre le site de l'Ouest et les trois sites de l'Est. L'analyse au niveau de l'infrapopulation a révélé un regroupement des génotypes de parasites au sein de chaque enfant, en particulier à Duekoué (Ouest) et Sikensi (Est). Parmi les six profils génétiques cox1-ITS2 possibles obtenus à partir de miracidia, S. bovis cox1 × S. haematobium ITS2 (42,0 %) était le plus fréquemment observé dans les populations. Nous avons identifié seulement 15 miracidia (0,7 %) avec un génotype S. bovis cox1 × S. bovis ITS2. Notre étude apporte de nouvelles connaissances sur la génétique des populations de S. haematobium et des hybrides S. haematobium × S. bovis chez l'homme en Côte d'Ivoire et nous plaidons pour la recherche de schistosomes hybrides chez les animaux (rongeurs et bovins) en Côte d'Ivoire.

Efficacy and safety of single-dose 40 mg/kg oral praziquantel in the treatment of schistosomiasis in preschool-age versus school-age children: An individual participant data meta-analysis.

BACKGROUND: Better knowledge of the efficacy and safety of single-dose 40 mg/kg oral praziquantel in preschool-age children is required, should preventive chemotherapy programs for schistosomiasis be expanded to include this age group. METHODOLOGY: We analyzed individual participant-level data from 16 studies (13 single-arm or cohort studies and three randomized trials), amounting to 683 preschool-age children (aged <6 years) and 2,010 school-age children (aged 6-14 years). Children had a documented Schistosoma mansoni or S. haematobium infection, were treated with single 40 mg/kg oral praziquantel, and assessed between 21 and 60 days post-treatment. Efficacy was expressed as arithmetic mean and individual egg reduction rate (ERR) and meta-analyzed using general linear models and mixed models. Safety was summarized using reported adverse events (AEs). PRINCIPAL FINDINGS: Preschool-age children had significantly lower baseline Schistosoma egg counts and more losses to follow-up compared to school-age children. No difference in efficacy was found between preschool- and school-age children using a general linear model of individual-participant ERR with baseline log-transformed egg count as covariate and study, age, and sex as fixed variables, and a mixed model with a random effect on the study. Safety was reported in only four studies (n = 1,128 individuals); few AEs were reported in preschool-age children 4 and 24 hours post-treatment as well as at follow-up. Three severe but not serious AEs were recorded in school-age children during follow-up. CONCLUSIONS/SIGNIFICANCE: There is no indication that single-dose 40 mg/kg oral praziquantel would be less efficacious and less safe in preschool-age children compared to school-age children, with the caveat that only few randomized comparisons exist between the two age groups. Preventive chemotherapy might therefore be extended to preschool-age children, with proper monitoring of its efficacy and safety.

Use of Insecticides in Agriculture and the Prevention of Vector-Borne Diseases: Population Knowledge, Attitudes, Practices and Beliefs in Elibou, South Côte d'Ivoire.

People's knowledge, attitudes, practices and beliefs (KAPB) pertaining to malaria are generally well described. However, little is known about population knowledge and awareness of insecticide resistance in malaria vectors. The aim of this study was to investigate KAPB related to insecticide resistance in malaria vectors due to the use of insecticides in agriculture and the prevention against mosquitoes. In mid-2017, we carried out a cross-sectional survey in Elibou, South Côte d'Ivoire, employing a mixed methods approach. Quantitative data were obtained with a questionnaire addressed to household heads. Interviews were conducted with key opinion leaders, including village chiefs, traditional healers, heads of health centres and pesticide sellers. Focus group discussions were conducted with youth and elders. A total of 203 individuals participated in the questionnaire survey (132 males, 65%). We found that people had good knowledge about malaria and mosquitoes transmitting the disease, while they felt that preventing measures were ineffective. Pesticides were intensively used by farmers, mainly during the rainy season. Among the pesticides used, insecticides and herbicides were most commonly used. While there was poor knowledge about resistance, the interviewees stated that insecticides were not killing the mosquitoes anymore. The main reason given was that insecticides were diluted by the manufacturers as a marketing strategy to sell larger quantities. More than a third of the farmers used agricultural pesticides for domestic purposes to kill weeds or mosquitoes. We observed a misuse of pesticides among farmers, explained by the lack of specific training. In the community, long-lasting insecticidal nets were the most common preventive measure against malaria, followed by mosquito coils and insecticide sprays. The interviewees felt that the most effective way of dealing with insecticide resistance was to combine at least two preventive measures. In conclusion, population attitudes and practices related to insecticides used in agriculture and the prevention against mosquitoes could lead to resistance in malaria vectors, while people's knowledge about insecticide resistance was limited. There is a need to raise awareness in communities about the presence of resistance in malaria vectors and to involve them in resistance management.

High prevalence of Schistosoma haematobium × Schistosoma bovis hybrids in schoolchildren in Côte d'Ivoire.

Schistosomiasis is a neglected tropical disease, though it is highly prevalent in many parts of sub-Saharan Africa. While Schistosoma haematobium-bovis hybrids have been reported in West Africa, no data about Schistosoma hybrids in humans are available from Côte d'Ivoire. This study aimed to identify and quantify S. haematobium-bovis hybrids among schoolchildren in four localities of Côte d'Ivoire. Urine samples were collected and examined by filtration to detect Schistosoma eggs. Eggs were hatched and 503 miracidia were individually collected and stored on Whatman® FTA cards for molecular analysis. Individual miracidia were molecularly characterized by analysis of mitochondrial cox1 and nuclear internal transcribed spacer 2 (ITS 2) DNA regions. A mitochondrial cox1-based diagnostic polymerase chain reaction was performed on 459 miracidia, with 239 (52.1%) exhibiting the typical band for S. haematobium and 220 (47.9%) the S. bovis band. The cox1 and ITS 2 amplicons were Sanger sequenced from 40 randomly selected miracidia to confirm species and hybrids status. Among the 33 cox1 sequences analysed, we identified 15 S. haematobium sequences (45.5%) belonging to seven haplotypes and 18 S. bovis sequences (54.5%) belonging to 12 haplotypes. Of 40 ITS 2 sequences analysed, 31 (77.5%) were assigned to pure S. haematobium, four (10.0%) to pure S. bovis and five (12.5%) to S. haematobium-bovis hybrids. Our findings suggest that S. haematobium-bovis hybrids are common in Côte d'Ivoire. Hence, intense prospection of domestic and wild animals is warranted to determine whether zoonotic transmission occurs.

Prevalence and Risk Factors for Schistosomiasis among Schoolchildren in two Settings of Côte d'Ivoire.

Schistosomiasis is a parasitic disease affecting more than 250 million people, primarily in sub-Saharan Africa. In Côte d'Ivoire both Schistosoma haematobium (causing urogenital schistosomiasis) and Schistosoma mansoni (causing intestinal schistosomiasis) co-exist. This study aimed to determine the prevalence of S. haematobium and S. mansoni and to identify risk factors among schoolchildren in the western and southern parts of Côte d'Ivoire. From January to April 2018, a cross-sectional study was carried out including 1187 schoolchildren aged 5-14 years. Urine samples were examined by a filtration method to identify and count S. haematobium eggs, while stool samples were subjected to duplicate Kato-Katz thick smears to quantify eggs of S. mansoni and soil-transmitted helminths. Data on sociodemographic, socioeconomic, and environmental factors were obtained using a pretested questionnaire. Multivariate logistic regression was employed to test for associations between variables. We found a prevalence of S. haematobium of 14.0% (166 of 1187 schoolchildren infected) and a prevalence of S. mansoni of 6.1% (66 of 1089 schoolchildren infected). In the southern part of Côte d'Ivoire, the prevalence of S. haematobium was 16.1% with a particularly high prevalence observed in Sikensi (35.6%), while S. mansoni was most prevalent in Agboville (11.2%). Swimming in open freshwater bodies was the main risk factor for S. haematobium infection (adjusted odds ratio (AOR) = 127.0, 95% confidence interval (CI): 25.0-634.0, p < 0.001). Fishing and washing clothes in open freshwater bodies were positively associated with S. haematobium and S. mansoni infection, respectively. Preventive chemotherapy using praziquantel should be combined with setting-specific information, education, and communication strategies in order to change children's behavior, thus avoiding contact with unprotected open freshwater.

Antagonistic effects of Plasmodium-helminth co-infections on malaria pathology in different population groups in Côte d'Ivoire.

INTRODUCTION: Plasmodium spp. and helminths are co-endemic in many parts of the tropics; hence, co-infection is a common phenomenon. Interactions between Plasmodium and helminth infections may alter the host's immune response and susceptibility and thus impact on morbidity. There is little information on the direction and magnitude of such interactions and results are conflicting. This study aimed at shedding new light on the potential interactions of Plasmodium and helminth co-infections on anemia and splenomegaly in different population groups in Côte d'Ivoire. METHODOLOGY: Parasitologic and clinical data were obtained from four cross-sectional community-based studies and a national school-based survey conducted between 2011 and 2013 in Côte d'Ivoire. Six scenarios of co-infection pairs defined as Plasmodium infection or high parasitemia, combined with one of three common helminth infections (i.e., Schistosoma mansoni, S. haematobium, and hookworm) served for analysis. Adjusted logistic regression models were built for each scenario and interaction measures on additive scale calculated according to Rothman et al., while an interaction term in the model served as multiplicative scale measure. PRINCIPAL FINDINGS: All identified significant interactions were of antagonistic nature but varied in magnitude and species combination. In study participants aged 5-18 years from community-based studies, Plasmodium-hookworm co-infection showed an antagonistic interaction on additive scale on splenomegaly, while Plasmodium-Schistosoma co-infection scenarios showed protective effects on multiplicative scale for anemia and splenomegaly in participants aged 5-16 years from a school-based study. CONCLUSIONS/SIGNIFICANCE: No exacerbation from co-infection with Plasmodium and helminths was observed, neither in participants aged 5-18 years nor in adults from the community-based studies. Future studies should unravel underlying mechanisms of the observed interactions, as this knowledge might help shaping control efforts against these diseases of poverty.

Are schoolchildren less infected if they have good knowledge about parasitic worms? A case study from rural Côte d'Ivoire.

BACKGROUND: Parasitic worms (helminths) are common infections in low- and middle-income countries. For most helminth species, school-aged children are at highest risk of infection and morbidity, such as impaired cognitive and physical development. Preventive chemotherapy is the current mainstay for helminthiases control. Sanitation improvement and hygiene-related education are important complementary strategies, which act by altering children's behaviour. However, little is known about the effect of improved knowledge on the risk of helminth infection. The aim of this study was to assess the potential influence of knowledge that children acquired at home or in school, without any specific health education intervention, on helminth infections. METHODS: In May 2014, we conducted a cross-sectional survey in western Côte d'Ivoire. A total of 2498 children, aged 9-12 years, were subjected to three consecutive stool examinations using duplicate Kato-Katz thick smears to determine infections with soil-transmitted helminths and Schistosoma mansoni. Additionally, children were interviewed to assess their knowledge about helminth infections. Four knowledge scores were constructed by factor analysis; one, reflecting general knowledge about helminths and three manifesting helminth species-specific knowledge. The effect of general and specific knowledge on children's helminth infection status was determined using meta-analysis. RESULTS: Children who scored high in the hookworm-specific knowledge were less likely to be infected with hookworm but no association was found for the other helminth species. Moreover, greater general knowledge was not associated with lower odds of being infected with any helminth species. Most of the children interviewed believed that the effect of preventive chemotherapy is permanent, and hence, re-treatment is not necessary. CONCLUSIONS: Specific knowledge about different types of helminths might not suffice to induce behavioural change which in turn reduces infection and reinfection with helminths. Health education interventions should strive to strengthen the perception of risk and to clarify the true benefit of preventive chemotherapy.

An Integrated Approach to Control Soil-Transmitted Helminthiasis, Schistosomiasis, Intestinal Protozoa Infection, and Diarrhea: Protocol for a Cluster Randomized Trial.

BACKGROUND: The global strategy to control helminthiases (schistosomiasis and soil-transmitted helminthiasis) emphasizes preventive chemotherapy. However, in the absence of access to clean water, improved sanitation, and adequate hygiene, reinfection after treatment can occur rapidly. Integrated approaches might be necessary to sustain the benefits of preventive chemotherapy and make progress toward interruption of helminthiases transmission. OBJECTIVE: The aim of this study was to assess and quantify the effect of an integrated control package that consists of preventive chemotherapy, community-led total sanitation, and health education on soil-transmitted helminthiasis, schistosomiasis, intestinal protozoa infection, and diarrhea in rural Côte d'Ivoire. METHODS: In a first step, a community health education program was developed that includes an animated cartoon to promote improved hygiene and health targeting school-aged children, coupled with a health education theater for the entire community. In a second step, a cluster randomized trial was implemented in 56 communities of south-central Côte d'Ivoire with 4 intervention arms: (1) preventive chemotherapy; (2) preventive chemotherapy plus community-led total sanitation; (3) preventive chemotherapy plus health education; and (4) all 3 interventions combined. Before implementation of the aforementioned interventions, a baseline parasitologic, anthropometric, and hygiene-related knowledge, attitudes, practices, and beliefs survey was conducted. These surveys were repeated 18 and 39 months after the baseline cross-sectional survey to determine the effect of different interventions on helminth and intestinal protozoa infection, nutritional indicators, and knowledge, attitudes, practices, and beliefs. Monitoring of diarrhea was done over a 24-month period at 2-week intervals, starting right after the baseline survey. RESULTS: Key results from this cluster randomized trial will shed light on the effect of integrated approaches consisting of preventive chemotherapy, community-led total sanitation, and health education against infections with soil-transmitted helminths, schistosomes, an intestinal protozoa and prevention of diarrhea in a rural part of Côte d'Ivoire. CONCLUSIONS: The research provided new insights into the acceptability, strengths, and limitations of an integrated community-based control package targeting helminthiases, intestinal protozoa infections, and diarrhea in rural communities of Côte d'Ivoire. In the longer term, the study will allow determining the effect of the integrated control approach on infection patterns with parasitic worms and intestinal protozoa, diarrheal incidence, anthropometric measures, and hygiene-related knowledge, attitudes, practices, and beliefs. TRIAL REGISTRATION: International Standard Randomized Controlled Trial Number (ISRCTN): 53102033; http://www.isrctn.com/ISRCTN53102033 (Archived by WebCite at http://www.webcitation.org/6wpnXEiHo). REGISTERED REPORT IDENTIFIER: RR1-10.2196/9166.

Epidemiology of intestinal parasite infections in three departments of south-central Côte d'Ivoire before the implementation of a cluster-randomised trial.

Hundreds of millions of people are infected with helminths and intestinal protozoa, particularly children in low- and middle-income countries. Preventive chemotherapy is the main strategy to control helminthiases. However, rapid re-infection occurs in settings where there is a lack of clean water, sanitation and hygiene. In August and September 2014, we conducted a cross-sectional epidemiological survey in 56 communities of three departments of south-central Côte d'Ivoire. Study participants were invited to provide stool and urine samples. Stool samples were examined for helminth and intestinal protozoa infections using the Kato-Katz technique and a formalin-ether concentration method. Urine samples were subjected to a filtration method for the diagnosis of Schistosoma haematobium. Information on sociodemographic characteristics, knowledge, attitude, practices and beliefs with regard to hygiene, sanitation and intestinal parasitic diseases were collected using a questionnaire administered to household heads. Multivariable logistic regression models were employed to analyse associations between parasite infections and risk factors. Overall, 4,305 participants had complete parasitological and questionnaire data. Hookworm was the predominant helminth species (21.2%), while Ascaris lumbricoides, Trichuris trichiura, Schistosoma mansoni and S. haematobium showed prevalences below 10%. Infections with pathogenic intestinal protozoa (e.g. Entamoeba histolytica/E. dispar and Giardia intestinalis) were similarly prevalent in the three departments. Hookworm infection was associated with open defecation and participants' age and sex. Entamoeba coli infection was negatively associated with the use of tap water at home (odds ratio (OR) = 0.66; p = 0.032). Disposal of garbage in close proximity to people's home was positively associated with G. intestinalis (OR = 1.30; p = 0.015). Taken together, helminth and intestinal protozoa infections affected a considerable proportion of rural dwellers in south-central Côte d'Ivoire at the onset of a cluster-randomised intervention trial. Our results will serve as baseline to monitor the effect of a package of interventions, including preventive chemotherapy, sanitation and health education on re-infection with helminths and intestinal protozoa. TRIAL REGISTRATION: ISRCTN53102033 (date assigned: 26 March 2014).

Effect of an integrated intervention package of preventive chemotherapy, community-led total sanitation and health education on the prevalence of helminth and intestinal protozoa infections in Côte d'Ivoire.

BACKGROUND: Preventive chemotherapy with donated anthelminthic drugs is the cornerstone for the control of helminthiases. However, reinfection can occur rapidly in the absence of clean water and sanitation coupled with unhygienic behaviour. The purpose of this study was to assess the effect of an integrated package of interventions, consisting of preventive chemotherapy, community-led total sanitation (CLTS) and health education, on the prevalence of helminth and intestinal protozoa infections and on participants' knowledge, attitude, practice and beliefs (KAPB) towards these diseases including water, sanitation and hygiene (WASH). METHODS: A cross-sectional survey was carried out in nine communities of south-central Côte d'Ivoire to assess people's infection with helminths and intestinal protozoa and KAPB. Subsequently, interventions were targeted to five communities, while the remaining communities served as control. The intervention encouraged latrine construction and an evaluation was done 6-7 months later to determine open defecation status of the respective communities. Anthelminthic treatment was provided to all community members. A follow-up cross-sectional survey was conducted approximately one year later, using the same procedures. RESULTS: Overall, 810 people had complete baseline and follow-up data and were given anthelminthic treatment. The baseline prevalence of hookworm, Schistosoma haematobium, Trichuris trichiura, Schistosoma mansoni and Ascaris lumbricoides was 31.1%, 7.0%, 2.0%, 1.0% and 0.3%, respectively. Four of the five intervention communities were classified open-defecation free. For hookworm infection, we observed higher negative changes in terms of proportion of decrease (-0.10; 95% confidence interval (CI): - 0.16, -0.04) and higher egg reduction rate (64.9 vs 15.2%) when comparing intervention with control communities. For intestinal protozoa, prevalence reduction was higher in intervention compared to control communities (8.2 vs 2.6%) and WASH indicators and intervention outcomes associated with lower odds for infection at follow-up. The intervention significantly impacted on reported latrine use (before: 15.5%, after: 94.6%), open defecation in the community surroundings (before: 75.0%, after: 16.7%) and awareness for environmental contamination through open defecation (before: 20.4%, after: 52.2%). CONCLUSIONS: An integrated package of interventions consisting of preventive chemotherapy, health education and CLTS reduces the prevalence of helminth and intestinal protozoa infection. Additional studies in other social-ecological settings are warranted to confirm our findings.

Estimating sensitivity of the Kato-Katz technique for the diagnosis of Schistosoma mansoni and hookworm in relation to infection intensity.

The Kato-Katz technique is the most widely used diagnostic method in epidemiologic surveys and drug efficacy trials pertaining to intestinal schistosomiasis and soil-transmitted helminthiasis. However, the sensitivity of the technique is low, particularly for the detection of light-intensity helminth infections. Examination of multiple stool samples reduces the diagnostic error; yet, most studies rely on a single Kato-Katz thick smear, thus underestimating infection prevalence. We present a model which estimates the sensitivity of the Kato-Katz technique in Schistosoma mansoni and hookworm, as a function of infection intensity for repeated stool sampling and provide estimates of the age-dependent 'true' prevalence. We find that the sensitivity for S. mansoni diagnosis is dominated by missed light infections, which have a low probability to be diagnosed correctly even through repeated sampling. The overall sensitivity strongly depends on the mean infection intensity. In particular at an intensity of 100 eggs per gram of stool (EPG), we estimate a sensitivity of 50% and 80% for one and two samples, respectively. At an infection intensity of 300 EPG, we estimate a sensitivity of 62% for one sample and 90% for two samples. The sensitivity for hookworm diagnosis is dominated by day-to-day variation with typical values for one, two, three, and four samples equal to 50%, 75%, 85%, and 95%, respectively, while it is only weakly dependent on the mean infection intensity in the population. We recommend taking at least two samples and estimate the 'true' prevalence of S. mansoni considering the dependence of the sensitivity on the mean infection intensity and the 'true' hookworm prevalence by taking into account the sensitivity given in the current study.

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

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

"Koko et les lunettes magiques": An educational entertainment tool to prevent parasitic worms and diarrheal diseases in Côte d'Ivoire.

BACKGROUND: Integrated control programs, emphasizing preventive chemotherapy along with health education, can reduce the incidence of soil-transmitted helminthiasis and schistosomiasis. The aim of this study was to develop an educational animated cartoon to improve school children's awareness regarding soil-transmitted helminthiasis, diarrheal diseases, and related hygiene practices in Côte d'Ivoire. The key messages included in the cartoon were identified through prior formative research to specifically address local knowledge gaps. METHODOLOGY: In a first step, preliminary research was conducted to assess the knowledge, attitudes, practices, and beliefs of school-aged children regarding parasitic worm infections and hygiene, to identify key health messages to be included in an animated cartoon. Second, an animated cartoon was produced, which included the drafting of the script and story board, and the production of the cartoon's initial version. Finally, the animated cartoon was pilot tested in eight selected schools and further fine-tuned. PRINCIPAL FINDINGS: According to the questionnaire results, children believed that the consumption of sweet food, eating without washing their hands, sitting on the floor, and eating spoiled food were the main causes of parasitic worm infections. Abdominal pain, diarrhea, lack of appetite, failure to grow, and general fatigue were mentioned as symptoms of parasitic worm infections. Most of the children knew that they should go to the hospital for treatment if they experienced symptoms of parasitic worm diseases. The animated cartoon titled "Koko et les lunettes magiques" was produced by Afrika Toon, in collaboration with a scientific team composed of epidemiologists, civil engineers, and social scientists, and the local school children and teachers. Pilot testing of the animated cartoon revealed that, in the short term, children grasped and kept key messages. Most of the children who were shown the cartoon reported to like it. Acceptance of the animated cartoon was high among children and teachers alike. The messaging was tailored to improve knowledge and practices for prevention of helminthiases and diarrheal diseases through prior identification of knowledge gaps. Integration of such education tools into the school curriculum, along with deworming campaigns, might improve sustainability of control and elimination efforts against helminthiases and diarrheal diseases.

A call to strengthen the global strategy against schistosomiasis and soil-transmitted helminthiasis: the time is now.

In 2001, the World Health Assembly (WHA) passed the landmark WHA 54.19 resolution for global scale-up of mass administration of anthelmintic drugs for morbidity control of schistosomiasis and soil-transmitted helminthiasis, which affect more than 1·5 billion of the world's poorest people. Since then, more than a decade of research and experience has yielded crucial knowledge on the control and elimination of these helminthiases. However, the global strategy has remained largely unchanged since the original 2001 WHA resolution and associated WHO guidelines on preventive chemotherapy. In this Personal View, we highlight recent advances that, taken together, support a call to revise the global strategy and guidelines for preventive chemotherapy and complementary interventions against schistosomiasis and soil-transmitted helminthiasis. These advances include the development of guidance that is specific to goals of morbidity control and elimination of transmission. We quantify the result of forgoing this opportunity by computing the yearly disease burden, mortality, and lost economic productivity associated with maintaining the status quo. Without change, we estimate that the population of sub-Saharan Africa will probably lose 2·3 million disability-adjusted life-years and US$3·5 billion of economic productivity every year, which is comparable to recent acute epidemics, including the 2014 Ebola and 2015 Zika epidemics. We propose that the time is now to strengthen the global strategy to address the substantial disease burden of schistosomiasis and soil-transmitted helminthiasis.

Spatial mapping and prediction of Plasmodium falciparum infection risk among school-aged children in Côte d'Ivoire.

BACKGROUND: In Côte d'Ivoire, malaria remains a major public health issue, and thus a priority to be tackled. The aim of this study was to identify spatially explicit indicators of Plasmodium falciparum infection among school-aged children and to undertake a model-based spatial prediction of P. falciparum infection risk using environmental predictors. METHODS: A cross-sectional survey was conducted, including parasitological examinations and interviews with more than 5,000 children from 93 schools across Côte d'Ivoire. A finger-prick blood sample was obtained from each child to determine Plasmodium species-specific infection and parasitaemia using Giemsa-stained thick and thin blood films. Household socioeconomic status was assessed through asset ownership and household characteristics. Children were interviewed for preventive measures against malaria. Environmental data were gathered from satellite images and digitized maps. A Bayesian geostatistical stochastic search variable selection procedure was employed to identify factors related to P. falciparum infection risk. Bayesian geostatistical logistic regression models were used to map the spatial distribution of P. falciparum infection and to predict the infection prevalence at non-sampled locations via Bayesian kriging. RESULTS: Complete data sets were available from 5,322 children aged 5-16 years across Côte d'Ivoire. P. falciparum was the predominant species (94.5 %). The Bayesian geostatistical variable selection procedure identified land cover and socioeconomic status as important predictors for infection risk with P. falciparum. Model-based prediction identified high P. falciparum infection risk in the north, central-east, south-east, west and south-west of Côte d'Ivoire. Low-risk areas were found in the south-eastern area close to Abidjan and the south-central and west-central part of the country. CONCLUSIONS: The P. falciparum infection risk and related uncertainty estimates for school-aged children in Côte d'Ivoire represent the most up-to-date malaria risk maps. These tools can be used for spatial targeting of malaria control interventions.

Bayesian risk profiling of soil-transmitted helminth infections and estimates of preventive chemotherapy for school-aged children in Côte d'Ivoire.

BACKGROUND: Soil-transmitted helminthiasis affects more than a billion people in the world and accounts for a global burden of 5.1 million disability-adjusted life years. The objectives of this study were (i) to map and predict the risk of soil-transmitted helminth infections among school-aged children in Côte d'Ivoire; (ii) to estimate school-aged children population-adjusted risk; and (iii) to estimate annual needs for preventive chemotherapy. METHODS: In late 2011/early 2012, a cross-sectional survey was carried out among school-aged children in 92 localities of Côte d'Ivoire. Children provided a single stool sample that was subjected to duplicate Kato-Katz thick smears for the diagnosis of soil-transmitted helminths. A Bayesian geostatistical variable selection approach was employed to identify environmental and socioeconomic risk factors for soil-transmitted helminth infections. Bayesian kriging was used to predict soil-transmitted helminth infections on a grid of 1 × 1 km spatial resolution. The number of school-aged children infected with soil-transmitted helminths and the amount of doses needed for preventive chemotherapy according to World Health Organization guidelines were estimated. RESULTS: Parasitological data were available from 5246 children aged 5-16 years. Helminth infections with hookworm were predominant (17.2 %). Ascaris lumbricoides and Trichuris trichiura were rarely found; overall prevalences were 1.9 % and 1.2 %, respectively. Bayesian geostatistical variable selection identified rural setting for hookworm, soil acidity and soil moisture for A. lumbricoides, and rainfall coefficient of variation for T. trichiura as main predictors of infection. The estimated school-aged children population-adjusted risk of soil-transmitted helminth infection in Côte d'Ivoire is 15.5 % (95 % confidence interval: 14.2-17.0 %). We estimate that approximately 1.3 million doses of albendazole or mebendazole are required for school-based preventive chemotherapy, and we provide school-aged children-adjusted risk aggregated at health district level. CONCLUSION: We provide the first soil-transmitted helminthiasis risk profile for entire Côte d'Ivoire, based on a robust Bayesian geostatistical framework. Our model-based estimates of treatment needs and risk maps on health district level may guide the national control program in spatial targeting of annual interventions.

The Effects of an Oil and Wheat Flour Fortification Program on Pre-School Children and Women of Reproductive Age Living in Côte d'Ivoire, a Malaria-Endemic Area.

Anemia and micronutrient deficiencies are widespread in sub-Saharan Africa, but the impact of food fortification is still debated. The objective of this study was to estimate the iron and vitamin A status of preschool children (PSC) and women of reproductive age (WRA) in households consuming fortified oil and wheat flour. The survey was cross-sectional in a rural and an urban area. Data on demographics, socioeconomic status, and fortified foods were collected at households. Hemoglobin (Hb), retinol binding protein (RBP), ferritin, soluble transferrin receptors (sTfR), subclinical inflammation, and Plasmodium spp. infection data were collected. In PSC, vitamin A deficiency (VAD) was prevalent, but for each 1 mg retinol equivalents (RE)/kg of oil consumed, RBP increased by 0.37 µmol/L (p = 0.03). In WRA, there was no significant VAD in the population (0.7%). Anemia was found in 92.2% of rural and 56.3% of urban PSC (p < 0.001). PSC with access to adequately fortified flour had Hb concentrations 15.7 g/L higher than those who did not (p < 0.001). Hb levels increased by +0.238 g/L per mg/kg increase in iron fortification levels (p < 0.001). The national program fortifying vegetable oil with vitamin A and wheat flour with iron and folic acid may have contributed to improved micronutrient status of PSC from two areas in Côte d'Ivoire.

New approaches to measuring anthelminthic drug efficacy: parasitological responses of childhood schistosome infections to treatment with praziquantel.

BACKGROUND: By 2020, the global health community aims to control and eliminate human helminthiases, including schistosomiasis in selected African countries, principally by preventive chemotherapy (PCT) through mass drug administration (MDA) of anthelminthics. Quantitative monitoring of anthelminthic responses is crucial for promptly detecting changes in efficacy, potentially indicative of emerging drug resistance. Statistical models offer a powerful means to delineate and compare efficacy among individuals, among groups of individuals and among populations. METHODS: We illustrate a variety of statistical frameworks that offer different levels of inference by analysing data from nine previous studies on egg counts collected from African children before and after administration of praziquantel. RESULTS: We quantify responses to praziquantel as egg reduction rates (ERRs), using different frameworks to estimate ERRs among population strata, as average responses, and within strata, as individual responses. We compare our model-based average ERRs to corresponding model-free estimates, using as reference the World Health Organization (WHO) 90% threshold of optimal efficacy. We estimate distributions of individual responses and summarize the variation among these responses as the fraction of ERRs falling below the WHO threshold. CONCLUSIONS: Generic models for evaluating responses to anthelminthics deepen our understanding of variation among populations, sub-populations and individuals. We discuss the future application of statistical modelling approaches for monitoring and evaluation of PCT programmes targeting human helminthiases in the context of the WHO 2020 control and elimination goals.

Use of an ecologically relevant modelling approach to improve remote sensing-based schistosomiasis risk profiling.

Schistosomiasis is a widespread water-based disease that puts close to 800 million people at risk of infection with more than 250 million infected, mainly in sub-Saharan Africa. Transmission is governed by the spatial distribution of specific freshwater snails that act as intermediate hosts and the frequency, duration and extent of human bodies exposed to infested water sources during human water contact. Remote sensing data have been utilized for spatially explicit risk profiling of schistosomiasis. Since schistosomiasis risk profiling based on remote sensing data inherits a conceptual drawback if school-based disease prevalence data are directly related to the remote sensing measurements extracted at the location of the school, because the disease transmission usually does not exactly occur at the school, we took the local environment around the schools into account by explicitly linking ecologically relevant environmental information of potential disease transmission sites to survey measurements of disease prevalence. Our models were validated at two sites with different landscapes in Côte d'Ivoire using high- and moderate-resolution remote sensing data based on random forest and partial least squares regression. We found that the ecologically relevant modelling approach explained up to 70% of the variation in Schistosoma infection prevalence and performed better compared to a purely pixel-based modelling approach. Furthermore, our study showed that model performance increased as a function of enlarging the school catchment area, confirming the hypothesis that suitable environments for schistosomiasis transmission rarely occur at the location of survey measurements.