Insecticide Resistance and Metabolic Mechanisms in Aedes aegypti from Two Agrosystems (Vegetable and Cotton Crops) in Côte d'Ivoire.

Background: The emergence of insecticide resistance in Aedes vectors mosquitoes poses a real challenge for arboviral-borne disease control. In Côte d'Ivoire, data are available on phenotypic resistance and the presence of kdr mutations in Aedes aegypti. Therefore, information on metabolic resistance in Aedes populations is very scarce. Here, we assessed the insecticide resistance status of Ae. aegypti in periurban and rural areas of Côte d'Ivoire, and we investigated the role of detoxification enzymes as possible resistance mechanisms. Materials and Methods: Aedes mosquito eggs were collected between June 2019 to April 2021 in two agricultural sites. Adults of Ae. aegypti were tested using World Health Organization tube assays, with seven insecticides belonging to pyrethroids, organochlorines, carbamates, and organophosphates classes. We determined the knockdown times (KdT50, KdT95) and resistance ratios of pyrethroids in natural populations. The synergist piperonyl butoxide (PBO) was used to investigate the role of enzymes in resistance. Biochemical assays were performed to detect potential increased activities in mixed-function oxidase levels, nonspecific esterases (NSEs), and glutathione S-transferases. Results: The results showed that Ae. aegypti populations were resistant to five insecticides with mortality of 46% and 89% for 0.75% permethrin, 68% and 92% for 0.05% deltamethrin, 57% and 89% for lambda-cyhalothrin, 41% and 47% for dichlorodiphenyltrichloroethane (DDT), 82% and 91% for chlorpyrifos-methyl in Songon-Agban and Kaforo, respectively. Susceptibility to carbamates was observed in our study sites. After exposure to PBO, the susceptibility of Ae. aegypti to pyrethroids and DDT was partially restored in Songon-Agban. Whereas in Kaforo, none increase of the mortality rates of Ae. aegypti for these four insecticides was observed after exposure to PBO. Increased activity of NSE (α-esterases) was found in Songon-Agban compared with the reference susceptible strain. Conclusion: These findings provide valuable information to support decisions for vector control strategies in Cote d'Ivoire. Also, we highlight the need for the monitoring of insecticide resistance management in Aedes vectors.

High vectorial transmission of malaria in urban and rural settings in the northern, western and eastern regions of Côte d'Ivoire.

BACKGROUND & OBJECTIVES: Malaria remains a public health problem in Côte d'Ivoire. To cope with this issue, the Ministry of Health established strategies through Long-Lasting Insecticidal Nets (LLINs) and artemisinin-based medicines. To better understand the influence of periodic mass distribution of LLINs on malaria transmission, this entomological survey was conducted in three regions of Côte d'Ivoire. METHODS: Mosquitoes were sampled by Human Landing Catches (HLC) in urban and rural settings of Korhogo, Man and Abengourou. Mosquitoes were identified morphologically and by molecular methods. Plasmodium falciparum infection was assessed by ELISA, and the Entomological Inoculation Rates (EIR) were calculated for each species and setting. RESULTS: Only An. gambiae s.l. was identified in Korhogo and in Abengourou while An. gambiae s.l. and An. funestus s.l. was reported in Man. An. coluzzii was the most abundant species of the An. gambiae siblings collected in Abengourou, and in Man while An. gambiae was most abundant in Korhogo. In urban settings, malaria vectors showed high aggressiveness (>11 bites per person per night) and the annual EIR was high (83.22-438.44 infectious bites per person per year). In rural settings, malaria vectors showed also high aggressiveness (19-52 b/p/n). The annual EIR is very high (>94 ib/p/yr). However, the weakest EIR was recorded in the northern region with 94.90 ib/p/yr. INTERPRETATION & CONCLUSION: This work indicates that malaria transmission remains high and heterogeneous across Côte d'Ivoire, despite repeated mass distribution of LLINs. Also, in Man, malaria transmission is more intense with the involvement of two main vectors. Furthermore, in the village of Korhogo, the EIR remained relatively low.

Mosquito (Diptera: Culicidae) Larval Ecology in Rubber Plantations and Rural Villages in Dabou (Côte d'Ivoire).

In Côte d'Ivoire, rubber cultivation has more than doubled since 2010. These mass agricultural areas require a large workforce with little information on how this environment might impact risk of mosquito-borne diseases. The objective of this study was to assess the larval ecology of mosquitoes in rubber areas of Dabou, Côte d'Ivoire. From January to June 2017, an entomological survey was conducted of mature (MP) and immature (IP) rubber plantations, as well as in villages surrounded by rubber plantations (SV) and remote from rubber plantations (RV). The number and type of potential and positive breeding sites were recorded, and mosquito larval densities and diversity were estimated. Seven genera divided into 31 species including major vector such as Anopheles gambiae s.l. and Aedes aegypti were identified. A total of 1,660 waterbodies were identified with a larvae positivity rate of 63.1%. A majority of waterbodies were identified in SV (N = 875, 53.4% positivity rate), followed by MP (N = 422, 81.8% positivity rate), IP (N = 194, 72.2% positivity rate) and least in RV (N = 169, 57.4% positivity rate). The most important breeding sites for disease vectors were leaf axils in IP (N = 108, 77.1%), latex collection cups in MP (N = 332, 96.2%) and the containers abandoned in the SV (N = 242, 51.8%) as well as in the RV (N = 59, 60.8%). All these results allow us to affirm that the cultivation of rubber trees has an impact on the larval ecology by increasing the number of available sites and favoring a high larval density and diversity.

Diversity of Anopheles Gambiae s.l., Giles (Diptera: Culicidae) Larval Habitats in Urban Areas and Malaria Transmission in Bouaké, Côte d'Ivoire.

Malaria vectors are supposedly uncommon in urban areas owing to the lack of suitable breeding sites for their development. However, the maintenance in urban areas of traditional rural practices along with humanitarian crisis can create favorable conditions for malaria transmission. This study aimed to provide relevant entomological data on the risk of malaria transmission in the city of Bouaké, after the military-political crisis from 2002 to 2011 in Côte d'Ivoire. Adults mosquitoes were collected by human landing catches in Dar Es Salam, Kennedy and N'gattakro neighborhoods. Potential breeding sites were georeferenced and mapped using a GPS. Mosquito species were identified morphologically and by molecular methods. Plasmodium infections were detected by quantitative PCR. Anopheline larvae were found in rice and vegetable crops, puddles, and footprints. A total of 939 Anopheles gambiae s.l. were caught during the surveys. The average human biting rate was 8.8 bites/person/night. The A. gambiae s.l. species were A. gambiae s.s (89.6%) and Anopheles coluzzii (10.4%). The average infectivity rate was 0.74% and the average annual entomological inoculation rate was estimated at 19 infected bites/person/year ranging from 0 in Dar Es Salam and N'gattakro to 58 in Kennedy. The risk of malaria transmission exists in Bouaké city, although Plasmodium infections are low.

Role of Anopheles gambiae s.s. and Anopheles coluzzii (Diptera: Culicidae) in Human Malaria Transmission in Rural Areas of Bouaké, in Côte d'Ivoire.

Studies done in Bouaké (Côte d'Ivoire) about 20-yr ago reported that Anopheles gambiae s.l. Giles was the major malaria vector. The present study aimed to update these data and to identify the main vectors. Mosquitoes were collected in Allokokro and Petessou villages between June 2014 and December 2015 using the human landing catching method. Potential breeding sites of An. gambiae s.l. were identified in August and October 2014 and mapped using GPS. Anopheles species were morphologically and molecularly [polymerase chain reaction (PCR)] identified. Ovaries of female were dissected to determine the parity and infection with Plasmodium was detected in head and thorax by quantitative PCR. In Allokokro, the biting rate of An. gambiae s.s was significantly greater than Anopheles coluzzii, whereas, in Petessou, biting rates of both species were comparable. Plasmodium falciparum (Haemosporida: Plasmodiidae), Plasmodium malariae (Haemosporida: Plasmodiidae), and Plasmodium ovale (Haemosporida: Plasmodiidae) identified in both villages. The infection rates of An. gambiae s.s. and An. coluzzii were not significantly different. The entomological inoculation rate (EIR) of An. gambiae s.s. for P. falciparum was 9-fold greater than that of An. coluzzii in Allokokro; however, in Petessou, the EIRs of both species were comparable. In both village, An. gambiae s.s was responsible for P. falciparum and P. ovale transmission whereas An. coluzzii transmitted all three Plasmodium species.

Vectorial Transmission of Malaria in Major Districts of Côte d'Ivoire.

To better understand the influence of periodic mass distribution of Long-Lasting Insecticidal Nets (LLINs) on malaria transmission, a 1-yr entomological survey was conducted in three major districts of Côte d'Ivoire. Mosquitoes were sampled by Human Landing Catches (HLC) in urban and rural areas of San Pedro and Abidjan (coastal), and in Yamoussoukro (central). Mosquitoes were identified morphologically and by molecular methods. The Plasmodium falciparum circumsporozoite (CSP) indices were measured by ELISA, and the Entomological Inoculation Rates (EIR) were calculated for each species and area. Anopheles gambiae s.l. Giles (Diptera: Culicidae) and Anopheles nili Theobald (Diptera: Culicidae) were identified in coastal districts, while An. gambiae s.l. and Anopheles funestus Giles (Diptera: Culicidae) were reported in the central district. In urban areas, malaria vectors showed a low aggressiveness (<10 bites per person per night), except in Yamoussoukro, where up to 18.9 b/p/n were recorded. The annual EIR was higher in the central urban area (138.7 infected bites per person per year) than in coastal ones (10-62 ib/p/n). In rural areas, malaria vectors were highly aggressive with an average 13 b/p/n for An. gambiae s.l, 21.2 b/p/n for An. nili and 12 b/p/n for An. funestus. The annual EIR ranged between 94.9 and 193.4 infected bites per person per year. This work indicates that, despite repeated mass distribution of LLINs, malaria transmission remains high and heterogeneous across Côte d'Ivoire. Malaria transmission was lower in coastal urban areas than in the central one, and remains high rural areas where two additional Anopheles vectors are involved in malaria transmission.

The current insecticide resistance status of Anopheles gambiae (s.l.) (Culicidae) in rural and urban areas of Bouaké, Côte d'Ivoire.

BACKGROUND: Several studies were carried out in experimental hut station in areas surrounding the city of Bouaké, after the crisis in Côte d'Ivoire. They reported increasing resistance levels to insecticide for malaria transmiting mosquitoes. The present work aims to evaluate the current resistance level of An. gambiae (s.l.) in rural and urban areas in the city of Bouaké. METHODS: Larvae of Anopheles gambiae (s.l.) were collected from five different study sites and reared to adult stages. The resistance status was assessed using the WHO bioassay test kits for adult mosquitoes, with eight insecticides belonging to pyrethroids, organochlorines, carbamates and organophosphates classes. Molecular assays were performed to identify the molecular forms of An. gambiae (s.l.), the L1014F kdr and the ace-1R alleles in individual mosquitoes. The synergist PBO was used to investigate the role of enzymes in resistance. Biochemical assays were performed to detect potential increased activities in mixed function oxidase (MFO) levels, non-specific esterases (NSE) and glutathione S-transferases (GST). RESULTS: High resistance levels to pyrethroids, organochlorines, and carbamates were observed in Anopheles gambiae (s.l.) from Bouaké. Mortalities ranged between 0 and 73% for the eight tested insecticides. The pre-exposure to PBO restored full or partial susceptibility to pyrethroids in the different sites. The same trend was observed with the carbamates in five sites, but to a lesser extent. With DDT, pre-exposure to PBO did not increase the mortality rate of An. gambiae (s.l.) from the same sites. Tolerance to organophosphates was observed. An increased activity of NSE and higher level of MFO were found compared to the Kisumu susceptible reference strain. Two molecular forms, S form [(An. gambiae (s.s)] and M form (An. coluzzi) were identified. The kdr allele frequencies vary from 85.9 to 99.8% for An. gambiae (s.s.) and from 81.7 to 99.6% for An. coluzzii. The ace-1R frequencies vary between 25.6 and 38.8% for An. gambiae (s.s.) and from 28.6 to 36.7% for An. coluzzii. CONCLUSION: Resistance to insecticides is widespread within both An. gambiae (s.s.) and An. coluzzii. Two mechanisms of resistance, i.e. metabolic and target-site mutation seemed to largely explain the high resistance level of mosquitoes in Bouaké. Pyrethroid resistance was found exclusively due to the metabolic mechanism.

Sustaining Control of Schistosomiasis Mansoni in Western Côte d'Ivoire: Results from a SCORE Study, One Year after Initial Praziquantel Administration.

BACKGROUND: The Schistosomiasis Consortium for Operational Research and Evaluation (SCORE) has launched several large-scale trials to determine the best strategies for gaining and sustaining control of schistosomiasis and transitioning toward elimination. In Côte d'Ivoire, a 5-year cluster-randomized trial is being implemented in 75 schools to sustain the control of schistosomiasis mansoni. We report Schistosoma mansoni infection levels in children one year after the initial school-based treatment (SBT) with praziquantel and compare with baseline results to determine the effect of the intervention. METHODOLOGY: The baseline cross-sectional survey was conducted in late 2011/early 2012 and the first follow-up in May 2013. Three consecutive stool samples were collected from 9- to 12-year-old children in 75 schools at baseline and 50 schools at follow-up. Stool samples were subjected to duplicate Kato-Katz thick smears. Directly observed treatment (DOT) coverage of the SBT was assessed and the prevalence and intensity of S. mansoni infection compared between baseline and follow-up. PRINCIPAL FINDINGS: The S. mansoni prevalence in the 75 schools surveyed at baseline was 22.1% (95% confidence interval (CI): 19.5-24.4%). The DOT coverage was 84.2%. In the 50 schools surveyed at baseline and one year after treatment, the overall prevalence of S. mansoni infection decreased significantly from 19.7% (95% CI: 18.5-20.8%) to 12.8% (95% CI: 11.9-13.8%), while the arithmetic mean S. mansoni eggs per gram of stool (EPG) among infected children slightly increased from 92.2 EPG (95% CI: 79.2-105.3 EPG) to 109.3 EPG (95% CI: 82.7-135.9 EPG). In two of the 50 schools, the prevalence increased significantly, despite a DOT coverage of >75%. CONCLUSIONS/SIGNIFICANCE: One year after the initial SBT, the S. mansoni prevalence had decreased. Despite this positive trend, an increase was observed in some schools. Moreover, the infection intensity among S. mansoni-infected children was slightly higher at the 1-year follow-up compared to the baseline situation. Our results emphasize the heterogeneity of transmission dynamics and provide a benchmark for the future yearly follow-up surveys of this multi-year SCORE intervention study.

Sustaining the Control of Schistosoma mansoni in Western Côte d'Ivoire: Baseline Findings Before the Implementation of a Randomized Trial.

We report baseline findings before the implementation of a 4-year intervention trial designed to assess the impact of three different school-based treatment schedules with praziquantel to sustain the control of intestinal schistosomiasis. The baseline survey was conducted in 75 schools of western Côte d'Ivoire previously identified with moderate Schistosoma mansoni endemicity (prevalence: 10-24% in children aged 13-14 years). Three stool samples collected over consecutive days were subjected to duplicate Kato-Katz thick smears each. A questionnaire was administered to collect village-specific information that is relevant for schistosomiasis transmission. Overall, 4,953 first graders (aged 5-8 years) and 7,011 school children (aged 9-12 years) had complete parasitologic data. The overall prevalence of S. mansoni was 5.4% among first graders and 22.1% in 9- to 12-year-old children. Open defecation was practiced in all villages. The current baseline findings will be important to better understand the dynamics of S. mansoni prevalence and intensity over the course of this trial that might be governed by village characteristics and specific treatment interventions.

The spatial distribution of Schistosoma mansoni infection in four regions of western Côte d'Ivoire.

Schistosomiasis poses a considerable public health burden in sub- Saharan Africa and a sound understanding of the spatial distribution facilitates to better target control interventions. The objectives of this study were i) to assess the prevalence of Schistosoma mansoni among school-aged children in four regions of western Côte d'Ivoire; ii) to determine demographic, climatic and environmental factors that influence the distribution of S. mansoni; and iii) to map and predict the distribution of S. mansoni in non-sampled locations. Parasitological surveys were carried out in 264 schools from June to December 2011. In each school, we aimed to examine 50 children for S. mansoni infection using duplicate Kato-Katz thick smears. Schools were georeferenced using a hand-held global positioning system receiver. Demographic data were obtained from readily available school lists, while climatic and environmental data were extracted from open-access remote sensing databases. Multivariable, binary non-spatial models and a Bayesian geostatistical logistic regression model were used to identify demographic, climatic and environmental risk factors for S. mansoni infection. Risk maps were developed based on observed S. mansoni prevalences and using Bayesian geostatistical models to predict prevalences at non-sampled locations. Overall, 12,462 children provided a sufficiently large stool sample to perform at least one Kato-Katz thick smear. The observed overall prevalence of S. mansoni infection was 39.9%, ranging from 0 to 100% at the unit of the school. Bayesian geostatistical analysis revealed that age, sex, altitude and difference between land surface temperature at day and night were significantly associated with S. mansoni infection. The S. mansoni risk map presented here is being been used by the national schistosomiasis control programme for spatial targeting of praziquantel and other interventions.

Sustaining control of schistosomiasis mansoni in moderate endemicity areas in western Côte d'Ivoire: a SCORE study protocol.

BACKGROUND: Schistosomiasis is a parasitic disease that occurs in the tropics and subtropics. The mainstay of control is preventive chemotherapy with praziquantel. In Africa, an estimated 230 million people require preventive chemotherapy. In western Côte d'Ivoire, infections with Schistosoma mansoni are widespread. To provide an evidence-base for programme decisions about preventive chemotherapy to sustain control of schistosomiasis, a 5-year multi-country study with different treatment arms has been designed by the Schistosomiasis Consortium for Operational Research and Evaluation (SCORE) and is currently being implemented in various African settings, including Côte d'Ivoire. METHODS/DESIGN: We report the study protocol, including ethics statement and insight from a large-scale eligibility survey carried out in four provinces in western Côte d'Ivoire. The study protocol has been approved by the ethics committees of Basel and Côte d'Ivoire. A total of 12,110 children, aged 13-14 years, from 264 villages were screened for S. mansoni using duplicate Kato-Katz thick smears from single stool samples. Among the schools with a S. mansoni prevalence of 10-24%, 75 schools were selected and randomly assigned to one of three treatment arms. In each school, three stool samples are being collected from 100 children aged 9-12 years annually and one stool sample from 100 first-year students at baseline and in the final year and subjected to duplicate Kato-Katz thick smears. Cost and coverage data for the different intervention arms, along with environmental, political and other characteristics that might impact on the infection prevalence and intensity will be recorded in each study year, using a pretested village inventory form. DISCUSSION: The study will document changes in S. mansoni infection prevalence and intensity according to different treatment schemes. Moreover, factors that determine the effectiveness of preventive chemotherapy will be identified. These factors will help to develop reasonable measures of force of transmission that can be used to make decisions about the most cost-effective means of lowering prevalence, intensity and transmission in a given setting. The gathered information and results will inform how to effectively sustain control of schistosomiasis at a low level in different social-ecological contexts. TRIAL REGISTRATION: ISRCTN99401114 (date assigned: 12 November 2014).

Immunomodulatory activity of polysaccharides isolated from Clerodendrum splendens: beneficial effects in experimental autoimmune encephalomyelitis.

BACKGROUND: Extracts of leaves from Clerodendrum have been used for centuries to treat a variety of medicinal problems in tropical Africa. However, little is known about the high-molecular weight active components conferring therapeutic properties to these extracts. METHODS: Polysaccharides from the leaves of Clerodendrum splendens were extracted and fractionated by ion exchange and size-exclusion chromatography. Molecular weight determination, sugar analysis, degree of methyl esterification, and other chemical characterization of the fractions were performed. Immunomodulatory activity of the fractions was evaluated by determining their ability to induce monocyte/macrophage nitric oxide (NO), cytokine production, and mitogen-activated protein kinase (MAPK) phosphorylation. Experimental autoimmune encephalomyelitis (EAE) was induced in C57BL/6 mice, and severity of EAE was monitored in mice treated with intraperitoneal (i.p.) injections of the most active polysaccharide fraction. Lymph nodes (LN) and spleen were harvested, and levels of cytokines in supernatants from LN cells and splenocytes challenged with myelin oligodendrocyte glycoprotein peptide were determined. RESULTS: Fractions containing type II arabinogalactan had potent immunomodulatory activity. Specifically, the high-molecular weight sub-fraction CSP-AU1 (average of 38.5 kDa) induced NO and cytokine [interleukin (IL)-1α, -1ß, -6, -10, tumor necrosis factor (TNF; designated previously as TNF-α), and granulocyte macrophage-colony stimulating factor (GM-CSF)] production by human peripheral blood mononuclear cells (PBMCs) and monocyte/macrophages. CSP-AU1-induced secretion of TNF was prevented by Toll-like receptor 4 (TLR4) antagonist LPS-RS, indicating a role for TLR4 signaling. Treatment with CSP-AU1 also induced phosphorylation of a number of MAPKs in human PBMC and activated AP-1/NF-κB. In vivo treatment of mice with CSP-AU1 and CSP-NU1 resulted in increased serum IL-6, IL-10, TNF, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1α/CCL3, and MIP-1ß/CCL4. CSP-AU1 treatment of mice with EAE (50 mg/kg, i.p., daily, 13 days) resulted in significantly reduced disease severity in this experimental model of multiple sclerosis. Levels of IL-13, TNF, interferon (IFN)-γ, IL-17, and GM-CSF were also significantly decreased, whereas transforming growth factor (TGF)-ß was increased in LN cells from CSP-AU1-treated EAE mice. CONCLUSIONS: Polysaccharide CSP-AU1 is a potent natural innate immunomodulator with a broad spectrum of agonist activity in vitro and immunosupressive properties after chronic administration in vivo.

Immunomodulatory and hemagglutinating activities of acidic polysaccharides isolated from Combretum racemosum.

Extracts of leaves of different species of the genus Combretum have been used historically to treat a variety of medicinal problems. However, little is known about the active components conferring therapeutic properties to these extracts. In the present studies, we evaluated biochemical properties and immunomodulatory activity of polysaccharides isolated from the leaves of Combretum racemosum. Water-soluble polysaccharides from leaves of C. racemosum were extracted and fractionated by DEAE-cellulose and Diaion HP-20 to obtain a Diaion-bound fraction, designated Combretum polysaccharide-acidic bound or CP-AB, which was eluted with methanol, and an unbound fraction, designated as CP-AU. Molecular weight determination, sugar analysis, and other physical and chemical characterization of the fractions were performed. Fraction CP-AU (mol. weight 5.0 kDa) contained type II arabinogalactan and had potent immunomodulatory activity, inducing the production of interleukin (IL)-1ß, -6, -10, and tumor necrosis factor-α (TNF-α) by human peripheral blood mononuclear cells (PBMC) and MonoMac-6 monocytic cells. Likewise, intraperitoneal administration of CP-AU increased in vivo serum levels of IL-6 and monocyte chemoattractant protein-1 (MCP-1) in mice. CP-AU-induced secretion of TNF-α in PBMC was prevented by Toll-like receptor 4 (TLR4) antagonist LPS-RS. Treatment with CP-AU induced phosphorylation of Akt2, Akt3, GSK-3ß, HSP27, mTOR, and all p38 MAPK isoforms (α, ß, δ, and γ), as well as stimulation of AP-1/NF-κB transcriptional activity. In addition, CP-AU effectively agglutinated erythrocytes from several species, including human, mouse, and rabbit. In contrast, fraction CP-AB was inactive in all biological tests, including cytokine production and hemagglutination. These data suggest that at least part of the beneficial therapeutic effects reported for the water extracts of leaves from C. racemosum are due to modulation of leukocyte functions.

Immunomodulatory activity of polysaccharides isolated from Alchornea cordifolia.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of leaves from different species of the genus Alchornea have been used for centuries to treat a variety of medicinal problems in tropical Africa. However, little is known about the high-molecular weight active components conferring therapeutic properties to these extracts. OBJECTIVE: The aim of this study was to evaluate the immunomodulatory activity of polysaccharides isolated from the leaves of Alchornea cordifolia. MATERIALS AND METHODS: Water-soluble polysaccharides from leaves of Alchornea cordifolia were extracted and fractionated by DEAE-cellulose, Diaion HP-20, and size-exclusion chromatography. Molecular weight, sugar analysis, and other physical and chemical characterization of the fractions were performed. Immunomodulatory activity of the polysaccharide fractions was evaluated by determining their ability to induce monocyte/macrophage nitric oxide (NO) and cytokine production. Activation of mitogen activated protein kinases (MAPK) was also assessed using a phospho-MAPK array. Activation of nuclear factor κB (NF-κB) was measured using an alkaline phosphatase reporter gene assay in THP1-Blue monocytic cells. RESULTS: Six polysaccharide fractions from Alchornea cordifolia were isolated. Fractions containing type II arabinogalactan had potent immunomodulatory activity. Particularly, the parent fraction AP-AU and its high-molecular weight sub-fraction AP-AU1 (average M(r) was estimated to be 39.5kDa) induced production of NO and cytokines [interleukin (IL)-1ß, -6, -10, tumor necrosis factor (TNF)-α, and granulocyte-macrophage-colony stimulating factor (GM-CSF)] in human peripheral blood mononuclear cells and human and murine monocyte/macrophages cell lines in vitro. Furthermore, treatment with AP-AU1 induced phosphorylation of Akt2, p38δ/p38γ, p70S6K1, RSK2, and mTOR, as well as stimulation of NF-κB transcriptional activity. CONCLUSION: Our results provide a molecular basis to explain a portion of the beneficial therapeutic properties of water extracts from Alchornea cordifolia leaves in traditional folk medicine of Africa.

Prevalence and spatial distribution of Entamoeba histolytica/dispar and Giardia lamblia among schoolchildren in Agboville area (Côte d'Ivoire).

BACKGROUND: New efforts are being made to improve understanding of the epidemiology of the helminths and intensifying the control efforts against these parasites. In contrast, relatively few studies are being carried out in this direction for the intestinal protozoa. To contribute to a better comprehension of the epidemiology of the intestinal protozoa, prevalence, and spatial distribution of Entamoeba histolytica/dispar and Giardia lamblia, and their association with drinking water supplies, were determined in the Agboville department in southeast Côte d'Ivoire. METHODS/FINDINGS: Stool samples were taken from more than 1,300 schoolchildren in the third year of primary education (CE1) from 30 primary schools and preserved in SAF (sodium acetate-acetic acid-formalin). The samples were analyzed by formalin-ether concentration. Then, a survey questionnaire addressed to schoolchildren and school directors was used to collect data on water supplies. Prevalence of E. histolytica/dispar and G. lamblia were, respectively, 18.8% and 13.9%. No particular focus zone was observed in the spatial distribution of the two species. Significant negative association was observed between use of tap water and high prevalence of E. histolytica/dispar infection (OR = 0.83, p = 0.01). High prevalence of G. lamblia infection was positively associated with use of ponds as the source of drinking water (OR = 1.28, p = 0.009). CONCLUSION: These two species of pathogenic protozoa are present with substantial prevalence in this area of Côte d'Ivoire. Although their spatial distribution is not focused in any one place, determination of the population segments with the highest levels of infection will help to target the chemotherapeutic fight. To reinforce treatment with chemotherapeutic agents, tap water should be made available in all the localities of this area.

Spatial risk profiling of Plasmodium falciparum parasitaemia in a high endemicity area in Côte d'Ivoire.

BACKGROUND: The objective of this study was to identify demographic, environmental and socioeconomic risk factors and spatial patterns of Plasmodium falciparum parasitaemia in a high endemicity area of Africa, and to specify how this information can facilitate improved malaria control at the district level. METHODS: A questionnaire was administered to about 4,000 schoolchildren in 55 schools in western Côte d'Ivoire to determine children's socioeconomic status and their habit of sleeping under bed nets. Environmental data were obtained from satellite images, digitized ground maps and a second questionnaire addressed to school directors. Finger prick blood samples were collected and P. falciparum parasitaemia determined under a microscope using standardized, quality-controlled methods. Bayesian variogram models were utilized for spatial risk modelling and mapping of P. falciparum parasitaemia at non-sampled locations, assuming stationary and non-stationary underlying spatial dependence. RESULTS: Two-thirds of the schoolchildren were infected with P. falciparum and the mean parasitaemia among infected children was 959 parasites/microl of blood. Age, socioeconomic status, not sleeping under a bed net, coverage rate with bed nets and environmental factors (e.g., normalized difference vegetation index, rainfall, land surface temperature and living in close proximity to standing water) were significantly associated with the risk of P. falciparum parasitaemia. After accounting for spatial correlation, age, bed net coverage, rainfall during the main malaria transmission season and distance to rivers remained significant covariates. CONCLUSION: It is argued that a massive increase in bed net coverage, particularly in villages in close proximity to rivers, in concert with other control measures, is necessary to bring malaria endemicity down to intermediate or low levels.

Spatially-explicit risk profiling of Plasmodium falciparum infections at a small scale: a geostatistical modelling approach.

BACKGROUND: There is a renewed political will and financial support to eradicate malaria. Spatially-explicit risk profiling will play an important role in this endeavour. Patterns of Plasmodium falciparum infection prevalence were examined among schoolchildren in a highly malaria-endemic area. METHODS: A questionnaire was administered and finger prick blood samples collected from 3,962 children, aged six to 16 years, attending 55 schools in a rural part of western Côte d'Ivoire. Information was gathered from the questionnaire on children's socioeconomic status and the use of bed nets for the prevention of malaria. Blood samples were processed with standardized, quality-controlled methods for diagnosis of Plasmodium spp. infections. Environmental data were obtained from satellite images and digitized maps. Bayesian variogram models for spatially-explicit risk modelling of P. falciparum infection prevalence were employed, assuming for stationary and non-stationary spatial processes. FINDINGS: The overall prevalence of P. falciparum infection was 64.9%, ranging between 34.0% and 91.9% at the unit of the school. Risk factors for a P. falciparum infection included age, socioeconomic status, not sleeping under a bed net, distance to health care facilities and a number of environmental features (i.e. normalized difference vegetation index, rainfall and distance to rivers). After taking into account spatial correlation only age remained significant. Non-stationary models performed better than stationary models. CONCLUSION: Spatial risk profiling of P. falciparum prevalence data provides a useful tool for targeting malaria control intervention, and hence will play a role in the quest of local elimination and ultimate eradication of the disease.

[Prevalence and polyparasitism of intestinal protozoa and spatial distribution of Entamoeba histolytica, E. dispar and Giardia intestinalis from pupils in the rural zone of Man in Côte d'Ivoire]. / Prévalences et polyparasitisme des protozoaires intestinaux et répartition spatiale d'Entamoeba histolytica/Entamoeba dispar et Giardia intestinalis chez des élèves en zone rurale de la région de Man en Côte-d'Ivoire.

Diseases caused by environmental contamination by micro-organisms, including intestinal helminths and protozoa, are prevalent in developing countries. According to some authors, their strong expansion in some zones of these countries is due primarily to favourable climatic conditions, combined with inadequate hygiene measures and cleaning and the generally low socio-economic level. Progress in disease control has resulted from new studies that improve our understanding of the epidemiology of helminthiases and from the availability of simple tools that are inexpensive and effective against these diseases (chemotherapy with albendazole and mebendazole). On the other hand, surprisingly few such studies have looked at intestinal protozoa, although the WHO reports that approximately 480 million individuals throughout the world are infested by amoebiasis caused by the protozoon Entamoeba histolytica and that 40,000-110,000 people die from it each year. Giardiasis, a cosmopolitan parasitosis, is due to another intestinal protozoon called Giardia intestinalis. To help develop a database on these parasites, we conducted a cross-sectional epidemiological survey in the Man region in western Côte d'Ivoire. Its objectives were to determine the prevalence of intestinal protozoa, to evaluate polyparasitism and to assess the spatial distribution of the pathogenic protozoal species, E. histolytica and G. intestinalis. Overall, 4466 stools samples taken from pupils aged 6 to 16 years of age at 57 different schools were analyzed under an optical microscope by the formol-ether stool concentration method, after preservation in sodium acetate-acetic acid-formalin (SAF). The most common protozoa species in this area were Endolimax nanus (83.8%) and E. coli (74.7%). The regional prevalence of G. intestinalis was 17.5% and of E. histolytica/E. dispar 11.3%. Both species were found in each of the 57 schools. The prevalence of E. histolytica/E. dispar exceeded 15% in six schools, and its spatial distribution was highly disparate. On the other hand, three large areas with substantial G. intestinalis transmission were identified. Polyparasitism of intestinal protozoa is very frequent in these areas; 80.2% of pupils carried at least 2 species. This work confirms the presence and breadth of protozoa species in the Man region. Understanding the spatial distribution of the principal areas where pathogenic protozoa species are transmitted may help to develop programmes for disease control that combine chemotherapy and preventive measures.

Disparities in parasitic infections, perceived ill health and access to health care among poorer and less poor schoolchildren of rural Côte d'Ivoire.

Differences in the state of health between rural and urban populations living in Africa have been described, yet only few studies analysed inequities within poor rural communities. We investigated disparities in parasitic infections, perceived ill health and access to formal health services among more than 4000 schoolchildren from 57 primary schools in a rural area of western Côte d'Ivoire, as measured by their socioeconomic status. In a first step, we carried out a cross-sectional parasitological survey. Stool specimens and finger prick blood samples were collected and processed with standardized, quality-controlled methods, for diagnosis of Schistosoma mansoni, soil-transmitted helminths, intestinal protozoa and Plasmodium. Then, a questionnaire survey was carried out for the appraisal of self-reported morbidity indicators, as well as housing characteristics and household assets ownership. Mean travel distance from each village to the nearest health care delivery structure was provided by the regional health authorities. Poorer schoolchildren showed a significantly higher infection prevalence of hookworm than better-off children. However, higher infection prevalences of intestinal protozoa (i.e. Blastocystis hominis, Endolimax nana and Iodamoeba butschlii) were found with increasing socioeconomic status. Significant negative associations were observed between socioeconomic status and light infection intensities with hookworm and S. mansoni, as well as with several self-reported morbidity indicators. The poorest school-attending children lived significantly further away from formal health services than their richer counterparts. Our study provides evidence for inequities among schoolchildren's parasitic infection status, perceived ill health and access to health care in a large rural part of Côte d'Ivoire. These findings call for more equity-balanced parasitic disease control interventions, which in turn might be an important strategy for poverty alleviation.

Cultivation and properties of Echinamoeba thermarum n. sp., an extremely thermophilic amoeba thriving in hot springs.

Here we describe a new, extremely thermophilic amoeba growing between 33 degrees C and 57 degrees C ( Topt.=50 degrees C). Isolates had been obtained from hot springs at Agnano Terme (Italy), Yellowstone National Park (USA), Kamchatka (Russia), and the Arenal Volcano (Costa Rica). They could be cultured monoxenically on a thermophilic alpha-proteobacterium. The morphology of the amoeba was studied using a microscope situated under a heatable polyacrylate hood. At 50 degrees C, the cells appeared flat with an irregular triangular or elongate shape, sometimes exhibiting fine spine-like subpseudopodia. On average, they were 22 microm long and 11 microm wide and had one nucleus with a central nucleolus. Based on morphology and on SSU rRNA comparisons, the amoeba belonged to the genus Echinamoeba, where it represents a new species. Referring to its extremely thermophilic lifestyle and its hydrothermal habitat, we name it E. thermarum.