Each patient underwent a standard clinical assessment and a questionnaire was completed. The CD4 cell count was assessed and up to three stool samples were collected and analyzed. Data collection Baseline characteristics regarding relevant demographics, place of residence and living conditions selleck chemicals (province of Laos, urban/rural housing, access to drinking water and in-house toilets, animal contact), clinical features (fever, weight loss, anorexia, diarrhea, nausea, vomiting, abdominal pain, and pulmonary, neurological and mucocutaneous symptoms) were collected at inclusion by the physician using a standardized questionnaire. According to World Health Organization definitions, diarrhea was defined as the passage of three or more loose or liquid stools per day [12].

It was considered acute when it lasted less than two weeks and persistent when it lasted 14 days or longer [12]. The immunological status of the patients related to HIV disease was assessed using the 2007 – revised World Health Organization HIV clinical staging and disease classification system [13]. Laboratory examination Serological diagnosis of HIV infection was made using two rapid screening tests: Determine HIV 1/2 (Inverness Medical, Tokyo, Japan) and Uni-Gold HIV (Trinity Biotech, Bray, Ireland). In the case of discordant results, the diagnosis was confirmed by the ELISA test Vironostika HIV Uniform II plus O (BioM��rieux, Marcy-l��Etoile, France). HIV viral load testing was not prescribed to patients with a newly diagnosed HIV infection because of limited availability of the technique.

The CD4 lymphocyte count was determined by flow cytometry (Cyflow SL and Cyflow Counter, Partec GmbH, Munster, Germany). Laboratory investigations for opportunistic infections were limited to direct diagnosis of mycobacterial infections and cryptococcosis using Ziehl-Neelsen stain on sputum samples and India ink test or antigen detection in cerebrospinal fluid samples, respectively. Stool sample analysis All patients were asked to provide three stool samples from three consecutive days. Each fresh stool specimen was processed as follows: the technicians of the hospital laboratories performed direct microscopic examination and examination of a Kato thick smear [5]. The sample was then preserved in 10% formalin and forwarded to the parasitological department of the Faculty of Medicine, University of Heath Sciences (Vientiane, Laos).

Batimastat The formalin-preserved samples were subjected to two techniques: formalin-ethyl concentration technique for protozoa and helminths diagnosis [14] and modified acid-fast staining for coccidia diagnosis (Cryptosporidium spp., Cyclospora cayetanensis, Cytoisospora belli, Sarcocystis hominis) [15]. The diagnoses were made with the assistance of parasitologists from the University of Lyon (France). Two smears from formalin-preserved samples were also prepared for microsporidia diagnosis.