ABSTRACT
The liver fluke Fasciola hepatica parasitizes humans and ruminant livestock worldwide, and it is now being considered a reemerging zoonotic disease, especially in areas in which it is endemic, such as South America. This study investigates the immune response to excretory and secretory products produced by F. hepatica in a group of patients from the Peruvian Altiplano, where the disease is highly endemic. Using a proteomic approach and immunoblotting techniques, we have identified the enzymes leucine aminopeptidase (LAP) and phosphoenolpyruvate carboxykinase as immunodominant antigens recognized by sera from fasciolosis patients. An indirect enzyme-linked immunosorbent assay using recombinant LAP as the antigen was developed to check sera from individuals of this region. Our results demonstrate that LAP produces a specific and strong reaction, suggesting its potential use in the serologic diagnosis of F. hepatica infections in humans.
Subject(s)
Antigens, Helminth/immunology , Fasciola hepatica/isolation & purification , Fascioliasis/immunology , Fascioliasis/veterinary , Immunodominant Epitopes/immunology , Leucyl Aminopeptidase/immunology , Sheep Diseases/parasitology , Adolescent , Animals , Antigens, Helminth/genetics , Antigens, Helminth/metabolism , Base Sequence , Blotting, Western/methods , Child , Child, Preschool , DNA Primers , Electrophoresis, Gel, Two-Dimensional/methods , Enzyme-Linked Immunosorbent Assay/methods , Fasciola hepatica/genetics , Fasciola hepatica/immunology , Fascioliasis/blood , Fascioliasis/parasitology , Humans , Immunodominant Epitopes/genetics , Immunodominant Epitopes/metabolism , Leucyl Aminopeptidase/genetics , Leucyl Aminopeptidase/metabolism , Molecular Sequence Data , Polymerase Chain Reaction/methods , Sheep , Sheep Diseases/immunologyABSTRACT
Twelve Spanish laboratories collected 325 yeast clinical isolates during a 30 day's period, among them 224 Candida albicans, 30 Candida glabrata, and 27 Candida parapsilosis. In vitro antifungal susceptibility to amphotericin B, ketoconazole, fluconazole and itraconazole was determined by an agar diffusion test (Neo-Sensitabs, Rosco, Denmark). All the isolates tested were susceptible in vitroto amphotericin B and nearly all (97.2%) to itraconazole. In vitrosusceptibility to fluconazole and ketoconazole was high (90.2% and 91.4% of isolates, respectively) but showed variations depending on the species tested. Resistance to fluconazole and ketoconazole was low in C. albicans (4% and 3%, respectively), but 30% of Candida guilliermondii and 36% of C. glabrata isolates were resistant to fluconazole. Ketoconazole resistance was observed in 40% of C. glabrata, and 17% of Candida tropicalis. Resistance to antifungal drugs is very low in Spain and it is related to non-C. albicans isolates.
