The disinfectant effects (DEs) of 10 types of chemicals, defined by their ability to destroy or inhibit oocysts and consequently prevent sporulation of Eimeria tenella field isolate, were evaluated in vitro. Correct species assignments and sample purities were confirmed by the singular internal transcribed spacer (ITS)-PCR analysis. A total of 18 treatments were performed, and the disinfection suppression levels were 75.9% for 39% benzene + 22% xylene (1:10 dilution), 85.5% for 30% cresol soup (1:1 dilution), and 91.7% for 99.9% acetic acid (1:2 dilution) group. The results indicate that acetic acid, cresol soup, and benzene+xylene are good candidates for suppression of E. tenella oocyst sporulation.
Animals ; Antiprotozoal Agents/*pharmacology ; Cluster Analysis ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal Spacer/chemistry/genetics ; Disinfectants/*pharmacology ; Eimeria tenella/*drug effects/*growth & development ; Microscopy ; Molecular Sequence Data ; Parasitic Sensitivity Tests ; Phylogeny ; Sequence Analysis, DNA ; Spores, Protozoan/*drug effects/*growth & development

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba.
Acanthamoeba castellanii/*growth & development/*metabolism ; Animals ; Gene Knockdown Techniques ; Lipid Metabolism ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/genetics ; RNA, Small Interfering/metabolism ; Rats ; Sequence Analysis, DNA ; Spores, Protozoan/*growth & development/*metabolism ; Ubiquitin-Conjugating Enzymes/genetics/*metabolism

A microsporidian parasite Enterocytozoon bieneusi is the most common microorganism recognized in AIDS patients, and slow scientific progress is attributed to our inability to propagate the parasite. We report upon the development of a system of propagation using the pig biliary system. The parasite spores were continuously detected in the bile samples post onset of spore shedding in the gall bladder, which suggests that this organism maintain persistent infection in the biliary system and that the hepatobiliary tree may represent a reservoir of infection. In conclusion the biliary tree is an adequate niche for the propagation of E. bieneusi. This work has also resulted in the development of a procedure of ultrasound-guided cholecystocentesis for aspirating biles. This is a simple and non-surgical procedure, and creates no signs of clinical complications in the livers and the gall bladders after dozens of separate attempts. Thus, this is a very useful and safe technique for the aspiration of bile from live animals.
AIDS-Related Opportunistic Infections/*parasitology ; Animals ; Bile/parasitology ; Biliary Tract/*parasitology ; DNA, Protozoan/analysis ; Disease Models, Animal ; Enterocytozoon/*growth & development/physiology ; Feces/parasitology ; Gallbladder/parasitology/ultrasonography ; Immunosuppression/veterinary ; Microsporidiosis/*parasitology ; Paracentesis/methods/veterinary ; Polymerase Chain Reaction/veterinary ; Specimen Handling/methods/veterinary ; Spores, Protozoan/isolation & purification/physiology ; Swine