Simple, Rapid Mycobacterium ulcerans Disease Diagnosis from Clinical Samples by Fluorescence of Mycolactone on Thin Layer Chromatography.

INTRODUCTION: Mycobacterium ulcerans infection, known as Buruli ulcer, is a disease of the skin and subcutaneous tissues which is an important but neglected tropical disease with its major impact in rural parts of West and Central Africa where facilities for diagnosis and management are poorly developed. We evaluated fluorescent thin layer chromatography (f-TLC) for detection of mycolactone in the laboratory using samples from patients with Buruli ulcer and patients with similar lesions that gave a negative result on PCR for the IS2404 repeat sequence of M. ulcerans. METHODOLOGY/PRINCIPAL FINDINGS: Mycolactone and DNA extracts from fine needle aspiration (FNA), swabs and biopsy specimen were used to determine the sensitivity and specificity of f-TLC when compared with PCR for the IS2404. For 71 IS2404 PCR positive and 28 PCR negative samples the sensitivity was 73.2% and specificity of 85.7% for f-TLC. The sensitivity was similar for swabs (73%), FNAs (75%) and biopsies (70%). CONCLUSIONS: We have shown that mycolactone can be detected from M. ulcerans infected skin tissue by f-TLC technique. The technique is simple, easy to perform and read with minimal costs. In this study it was undertaken by a member of the group from each endemic country. It is a potentially implementable tool at the district level after evaluation in larger field studies.

[Intestinal parasitoses in a village of Côte d'Ivoire. II: Relationship between intestinal parasitoses, physical and physiological development and child nutrition]. / Les parasitoses intestinales dans un village de Côte d'Ivoire. II: relations entre le parasitisme intestinal et le développement physique, physiologique et l'état de nutrition des enfants.

In a village situated at the border of the Ebrié lagoon, with economical problems, 343 school and preschool children, aged 4 to 15 (195 boys and 148 girls), considered by the school and their families to be in good health, were submitted for clinical and coproparasitologic examination. Basic clinical tests, anthropometric examination, spleen rate, nutrition status assessment (brachial perimeter of children aged 4 to 10 years) and the puberty level evaluation (using Tanner's scale) were independently completed. Splenomegaly rate among children between 4 and 9 years old was particularly important (78.0 %). Intestinal parasites were found in 84.8% of cases during the first examination (70.4% of children were diagnosed with multiple parasitosis). The most frequent parasite identified was Ascaris lumbricoides, being in 62.1 % of the cases, with a parasitic intensity between 1.000 and 52.000 OPG and worm burden between 1 and 16. According to the age, the highest prevalence of A. lumbricoides was higher (78.6 %) in children between 7 and 10 years. Results of anthropometric data, interpreted using NCHS percentile chart, indicated a significant stature-ponderal underdevelopment. Thus, concerning the height, 22.7% of all children were below normal value (below the 5th percentile), 74.3% between normal values and only 2.9% were above the 95th percentile. Among the 255 children with values between the normal limits, 73.7% (more than two thirds) were in the lower half, below the 50th percentile and only 26.3% (less than one third) were between the 50th and 95th percentile. According to the weight, 20.7% of children were below the normal value (below the 5th percentile), 77.8% had normal weight and 1.5% were above the 95th percentile. Even among children with normal weight, two thirds were in the lower half, below the 50th percentile. The underdevelopment was more evident after 9 years of age, particularly in girls. This coincided with higher rates of infection with roundworms. Regarding the nutritional status, 76.9% of the boys and 77.2% of the girls had a lower brachial perimeter than normal data, most frequently by one or two centimeters. A deficit of 3 to 4 cm. was found in 16.2% of the children, and it was higher among girls. The evaluation of pubertal development highlighted a certain delay in girls aged 10 to 12, with a preadolescence state of 76.0 % at age 10, 20.0 % at age 11 and 15.8 % at age 12. Between 13 and 15 years of age, all girls began pubertal development, but none reached Tanner's stage 5. It is concluded that the highest intestinal parasitism (particularly roundworms) can have a negative effect on the staturo-ponderal development of children, their nutrition status and pubertal development. Paludism and alimentary deficiencies have to be also considered.

[Intestinal parasitoses in a village of Côte d'Ivoire. I: Control and prevention plan]. / Les parasitoses intestinales dans un village de Côte d'Ivoire. I: essai de mise en place d'une stratégie de lutte et de prévention.

The goal was to develop a complex medical, hygienic, sanitary and educational plan for control and prevention of intestinal parasitic infections in the rural areas in Ivory Coast. In a village situated at the border of the Ebrié lagoon, 416 persons were examined: 371 children, of which 343 were school and preschool children, aged 4 to 15 years (195 boys and 148 girls), 28 young children aged 6 months to 3 years, and a group of 45 adults. The parasitologic exams included perianal swabs (Graham's method), stool examination using saline solution, iodized solution (Lugol) and preparation Kato-Miura's method in thick layer. Parasitic intensity was done for helminths and worm burden have been carried after specific treatment of roundworms. Hygienic conditions as environment, school, dwelling and personal hygiene, eating habits, drinking water sanitation, garbage disposal, toilets, reproduction areas of hematophagous and mechanical vectors etc. have recorded. The prevalence of intestinal parasites was 84.8% in children (with 76.7 % polyparasites) and 29.0 % in adults. The results pointed out a hyperendemic zone. Parasitic infectious transmitted from person to person was frequent among children: 37.3% pinworms in school children, 30.3% amoeba cysts and 30.3% flagellate. Infections transmitted by soil were predominant, with 62.1 % roundworms (78.6 % in children aged 7 to 10 years) presenting an important parasitic intensity and worm burden. The parasitoses transmitted as larvae were frequent, only Strongyloides stercoralis being most frequent parasite in adults compared to children. A feasible plan of control the intestinal parasites has been established in collaboration with the local hospital, village leaders and health workers. Short-term measures have been carefully chosen, targeting especially the schools, teachers and health workers. The first health education measure concerns the hand cleanliness at home and at schools. It was suggested that a bucket of water be used per class, that the water be changed more often during the day, and soap be made available at all time. Lessons on the ways of transmission of parasites will be introduced in schools. A door-to-door education plan was discussed with village health workers and hospital nurses and laboratory technicians during the maternal-infantile prophylactic visits. The health education problems have been discussed extensively with village health workers. As a preliminary example, the prevention and campaign against the pinworm, a common parasite in children was chosen, whose transmission mechanism from person to person can be easily understood by children and mothers. Simultaneously the prevention of parasitic infections contributes extensively to the prevention of other serious diseases, as the typhoid fever etc. which are endemic in the region. Long-term preventive measures have been discussed with village leaders. The first measure is to fix the deep-well drinking water pump station of the village, financed by outside parties, with labour provided by the village. Measures for proper maintenance of the water pump station have also been discussed with representatives of the village. The program of the World Health Organization and National Institute of Hygiene of Ivory Coast concerning the periodic treatment of intestinal helminths, especially A. lumbricoides, given to all school aged children was discussed.