Développement d’une méthode PCR-SSCP pour caractériser le régime alimentaire des gastéropodes

Déterminer le régime alimentaire des gastéropodes terrestres est important étant donné leurs impacts écologiques et économiques. L’observation en nature est difficile, car les gastéropodes sont principalement actifs la nuit et sont de petite taille. De plus, l’analyse visuelle des contenus digestifs est compliquée car les gastéropodes broient leurs aliments, ne permettant pas une identification précise. L’objectif est de produire une méthode fiable et peu onéreuse pour déterminer le régime alimentaire des gastéropodes en milieu naturel. Le modèle, Arion fuscus, a été échantillonné au printemps dans quatre érablières anciennes du sud du Québec (Canada). Une approche basée sur l’ADN a été utilisée avec amplification par PCR d’un segment d’un gène chloroplastique. La diversité a été déterminée par électrophorèse sur gel non-dénaturant suivie d’un séquençage des variants d’intérêts pour identifier les plantes consommées. Pour les 52 limaces analysées, les PCR ont fonctionné avec succès et les extractions d’ADN de mêmes individus traités indépendamment ont montré les mêmes résultats sur les gels SSCP. Des résultats fiables ont été produits en quelques semaines et le nombre d’échantillons à séquencer a été limité, réduisant ainsi les coûts. Les résultats montrent une faible diversité végétale consommée (6 haplotypes) par A. fuscus; 78,8% des limaces ont uniquement consommé la même espèce de plante. Cette méthode peut permettre de meilleurs suivis des espèces consommées par les gastéropodes dans un contexte d’écologie de la conservation ou de contrôle des espèces nuisibles.The diet of terrestrial gastropods is difficult to assess in nature because of their often-nocturnal activity and their small size. Moreover, visual analyzes of digestive contents are problematic because gastropods grind food into small particles, making visual identification unexploitable. Determining the gastropods ‘diet is nevertheless highly relevant because gastropods may have strong ecological and economic impacts. On the one hand, several species are listed endangered or critically endangered. On the opposite, numerous species are considered agricultural pests. The objective is to produce an accurate and inexpensive method to determine the diet of gastropods. More specifically, by using an approach based on PCR-amplified DNA followed by SSCP gels, it was possible to determine taxa diversity before Sanger sequencing, and thus reduce costs. We used Arion fuscus from four old maple forests sites in southern Quebec (Canada) as model to test the method. The 52 individuals’ digestive contents analyzed resulted in a successful PCR amplification and independent DNA extractions from a given individual gave the same SSCP pattern. This method produced reliable results in a few weeks and at a low cost, making it possible to sequence only a few samples to know the plant species consumed by all individuals in the study. Results revealed a low diversity of plants consumed (6 haplotypes). 78.8% of the slugs harvested in spring consumed one and the same species of plant. This method enables the monitoring of plant species consumed by gastropods in conservation and agricultural pest control

Atelier paludisme: an international malaria training course held in Madagascar

The Atelier Paludisme (Malaria Workshop) is an international training course organized by the Institut Pasteur de Madagascar, which has been held annually for the past five years. The course was designed for both young and experienced researchers, as well as for healthcare professionals, mostly from malaria-endemic countries. Its objective is to provide participants with a broad knowledge of all features of malaria, to improve their skills in project management, to break geographical isolation by using the Internet as a source of documentary information. This six-week course makes use of concepts of andragogy and problem-based learning, i.e. a relationship between participants and tutors, which promotes a process of exchange rather than the simple transmission of knowledge, where participants have to search actively for information. This approach to training, combined with the wide background and experience of those involved, creates positive dynamics and enables participants to acquire new skills, develop their critical and analytical abilities. This paper describes the course and the lessons learned from its evaluation

Conditions de travail de jeunes serveuses dans le quartier touristique de Hô Chi Minh Ville : une analyse des rapports sociaux de sexe

Tableau d’honneur de la Faculté des études supérieures et postdoctorales, 2009-2010Le présent mémoire décrit les conditions de travail de jeunes serveuses à Phạm Ngũ Lạo, quartier touristique de Hồ Chí Minh Ville, au Việt Nam. Les serveuses, sans constituer un groupe homogène, vivent toutes une surcharge de travail, une grande fatigue, un stress intense face à la maladie et à la faiblesse de leurs revenus. Ces conditions affectent leur santé et leur bien-être. Ce mémoire analyse, dans une perspective féministe, les relations inégalitaires des serveuses avec leurs collègues masculins, leurs patrons et leurs clients. Ces relations, marquées par une forte différence de classe, sont complexifiées par la présence de nombreux étrangers parmi la clientèle. Fréquent, le harcèlement sexuel est omniprésent dans certains établissements. Les serveuses, confrontées à la pauvreté et à la précarité, courent le risque d’être entraînées vers la prostitution. Pour échapper à ce piège et améliorer leurs conditions, elles développent avec plus ou moins de succès diverses tactiques et stratégies. Mots-clés : serveuse, restauration, pauvreté, rapports sociaux de sexe, de classe et de race, genre et développement, quartier touristique, tourisme, tourisme sexuel, prostitution, harcèlement sexuel, tactiques et stratégies.This thesis describes the working conditions of young waitresses in Phạm Ngũ Lạo, a touristic neighbourhood in Hồ Chí Minh City, Việt Nam. Waitresses, without constituting a homogenous group, are often subjected to heavy workloads and low earnings, which can lead to an intensification of stress, fatigue, and illness. The working conditions affect their health and well-being. This thesis analyzes, with a feminist perspective, the unequal relations between waitresses and their male counterparts, their superiors, and their clients. Those relations, clearly shaped by deep class disparities, are complexified by the presence of a foreign clientele. Sexual harassment is omnipresent in some establishments. Waitresses, confronted with poverty and insecurity, risk being drawn into prostitution. In order to escape this trap and to improve their conditions, they develop various tactics and strategies with mitigated outcome. Keywords: waitress, restauration, poverty, gender, class and race relations, gender and development, touristic neighbourhood, tourism, sexual tourism, prostitution, sexual harassment, tactics and strategies

Low-technology cooling box for storage of malaria RDTs and other medical supplies in remote areas

<p>Abstract</p> <p>Background</p> <p>With the increase in use of point-of-care diagnostic tests for malaria and other diseases comes the necessity of storing the diagnostic kits and the drugs required for subsequent management, in remote areas, where temperatures are high and electricity supply is unreliable or unavailable.</p> <p>Methods</p> <p>To address the lack of temperature-controlled storage during the introduction of community-based malaria management in Cambodia, the Cambodian National Centre for Parasitology, Entomology and Malaria Control (CNM) developed prototype evaporative cooling boxes (Cambodian Cooler Boxes - CCBs) for storage of perishable medical commodities in remote clinics. The performance of these CCBs for maintaining suitable storage temperatures was evaluated over two phases in 2005 and 2006-7, comparing conditions in CCBs using water as designed, CCBs with no water for evaporation, and ambient storage room temperatures. Temperature and humidity was monitored, together with the capacity of the RDTs recommended for storage between 2 to 30 degree Celsius to detect low-density malaria parasite samples after storage under these conditions.</p> <p>Results</p> <p>Significant differences were recorded between the proportion of temperatures within the recommended RDT storage conditions in the CCBs with water and the temperatures in the storage room (p < 0.001) and maximum temperatures were lower. RDTs stored at ambient temperatures were negative when tested with parasitized blood (2,000 parasites per micro litre) at 210 days, while the field RDTs kept in CCBs with water gave positive results until 360 days.</p> <p>Discussion and Conclusions</p> <p>The CCB was an effective tool for storage of RDTs at optimal conditions, and extended the effective life-span of the tests. The concept of evaporative cooling has potential to greatly enhance access to perishable diagnostics and medicines in remote communities, as it allows prolonged storage at low cost using locally-available materials, in the absence of electricity.</p

A possible alternative method for collecting mosquito larvae in rice fields

BACKGROUND: Rice fields are efficient breeding places for malaria vectors in Madagascar. In order to establish as easily as possible if a rice field is an effective larval site for anophelines, we compared classical dipping versus a net as methods of collecting larvae. RESULTS: Using similar collecting procedures, we found that the total number of anopheline larvae collected with the net was exactly double (174/87) that collected by dipping. The number of anopheline species collected was also greater with a net. CONCLUSIONS: The net is an effective means of collecting anopheline larvae and can be used for qualitative ecological studies and to rapidly determine which rice fields are containing malaria vectors

Plasmodium falciparum parasite population structure and gene flow associated to anti-malarial drugs resistance in Cambodia

Background: Western Cambodia is recognized as the epicentre of emergence of Plasmodium falciparum multi-drug resistance. The emergence of artemisinin resistance has been observed in this area since 2008–2009 and molecular signatures associated to artemisinin resistance have been characterized in k13 gene. At present, one of the major threats faced, is the possible spread of Asian artemisinin resistant parasites over the world threatening millions of people and jeopardizing malaria elimination programme efforts. To anticipate the diffusion of artemisinin resistance, the identification of the P. falciparum population structure and the gene flow among the parasite population in Cambodia are essential. Methods: To this end, a mid-throughput PCR-LDR-FMA approach based on LUMINEX technology was developed to screen for genetic barcode in 533 blood samples collected in 2010–2011 from 16 health centres in malaria endemics areas in Cambodia. Results: Based on successful typing of 282 samples, subpopulations were characterized along the borders of the country. Each 11-loci barcode provides evidence supporting allele distribution gradient related to subpopulations and gene flow. The 11-loci barcode successfully identifies recently emerging parasite subpopulations in western Cambodia that are associated with the C580Y dominant allele for artemisinin resistance in k13 gene. A subpopulation was identified in northern Cambodia that was associated to artemisinin (R539T resistant allele of k13 gene) and mefloquine resistance. Conclusions: The gene flow between these subpopulations might have driven the spread of artemisinin resistance over Cambodia

A microarray-based system for the simultaneous analysis of single nucleotide polymorphisms in human genes involved in the metabolism of anti-malarial drugs

Background: In order to provide a cost-effective tool to analyse pharmacogenetic markers in malaria treatment, DNA microarray technology was compared with sequencing of polymerase chain reaction (PCR) fragments to detect single nucleotide polymorphisms (SNPs) in a larger number of samples. Methods: The microarray was developed to affordably generate SNP data of genes encoding the human cytochrome P450 enzyme family (CYP) and N-acetyltransferase-2 (NAT2) involved in antimalarial drug metabolisms and with known polymorphisms, i.e. CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, and NAT2. Results: For some SNPs, i.e. CYP2A6*2, CYP2B6*5, CYP2C8*3, CYP2C9*3/*5, CYP2C19*3, CYP2D6*4 and NAT2*6/*7/*14, agreement between both techniques ranged from substantial to almost perfect (kappa index between 0.61 and 1.00), whilst for other SNPs a large variability from slight to substantial agreement (kappa index between 0.39 and 1.00) was found, e. g. CYP2D6*17 (2850C&gt;T), CYP3A4*1B and CYP3A5*3. Conclusion: The major limit of the microarray technology for this purpose was lack of robustness and with a large number of missing data or with incorrect specificity

Residual Antimalarial Concentrations before Treatment in Patients with Malaria from Cambodia: Indication of Drug Pressure

Background. The Thai-Cambodian border has been known as the origin of antimalarial drug resistance for the past 30 years. There is a highly diverse market for antimalarials in this area, and improved knowledge of drug pressure would be useful to target interventions aimed at reducing inappropriate drug use. Methods. Baseline samples from 125 patients with falciparum malaria recruited for 2 in vivo studies (in Preah Vihear and Pursat provinces) were analyzed for the presence of 14 antimalarials in a single run, by means of a liquid chromatography-tandem mass spectrometry assay. Results. Half of the patients had residual drug concentrations above the lower limit of calibration for at least 1 antimalarial at admission. Among the drugs detected were the currently used first-line drugs mefloquine (25% and 35% of patients) and piperaquine (15% of patients); the first-line drug against vivax malaria, chloroquine (25% and 41% of patients); and the former first-line drug, quinine (5% and 34% patients). Conclusions. The findings demonstrate that there is high drug pressure and that many people still seek treatment in the private and informal sector, where appropriate treatment is not guaranteed. Promotion of comprehensive behavioral change, communication, community-based mobilization, and advocacy are vital to contain the emergence and spread of parasite resistance against new antimalarial

Artemisinin resistance in Plasmodium falciparum malaria.

BACKGROUND: Artemisinin-based combination therapies are the recommended first-line treatments of falciparum malaria in all countries with endemic disease. There are recent concerns that the efficacy of such therapies has declined on the Thai-Cambodian border, historically a site of emerging antimalarial-drug resistance. METHODS: In two open-label, randomized trials, we compared the efficacies of two treatments for uncomplicated falciparum malaria in Pailin, western Cambodia, and Wang Pha, northwestern Thailand: oral artesunate given at a dose of 2 mg per kilogram of body weight per day, for 7 days, and artesunate given at a dose of 4 mg per kilogram per day, for 3 days, followed by mefloquine at two doses totaling 25 mg per kilogram. We assessed in vitro and in vivo Plasmodium falciparum susceptibility, artesunate pharmacokinetics, and molecular markers of resistance. RESULTS: We studied 40 patients in each of the two locations. The overall median parasite clearance times were 84 hours (interquartile range, 60 to 96) in Pailin and 48 hours (interquartile range, 36 to 66) in Wang Pha (P<0.001). Recrudescence confirmed by means of polymerase-chain-reaction assay occurred in 6 of 20 patients (30%) receiving artesunate monotherapy and 1 of 20 (5%) receiving artesunate-mefloquine therapy in Pailin, as compared with 2 of 20 (10%) and 1 of 20 (5%), respectively, in Wang Pha (P=0.31). These markedly different parasitologic responses were not explained by differences in age, artesunate or dihydroartemisinin pharmacokinetics, results of isotopic in vitro sensitivity tests, or putative molecular correlates of P. falciparum drug resistance (mutations or amplifications of the gene encoding a multidrug resistance protein [PfMDR1] or mutations in the gene encoding sarco-endoplasmic reticulum calcium ATPase6 [PfSERCA]). Adverse events were mild and did not differ significantly between the two treatment groups. CONCLUSIONS: P. falciparum has reduced in vivo susceptibility to artesunate in western Cambodia as compared with northwestern Thailand. Resistance is characterized by slow parasite clearance in vivo without corresponding reductions on conventional in vitro susceptibility testing. Containment measures are urgently needed. (ClinicalTrials.gov number, NCT00493363, and Current Controlled Trials number, ISRCTN64835265.