COMPARATIVE ANALYSIS OF STRESS AGENTS IN A SIMPLIFIED IN VITRO SYSTEM OF NEOSPORA CANINUM BRADYZOITE PRODUCTION

• Published in: The Journal of parasitology Vol. 90; no. 3; pp. 466 - 470
• Main Authors: Risco-Castillo, V, Fernández-García, A, Ortega-Mora, L. M
• Format: Journal Article
• Language: English
• Published: Lawrence, KS American Society of Parasitologists 01.06.2004; Allen Press Inc
• Subjects: abiotic stress; Animal diseases; Animal species; Animals; Antibodies; Antigens; Antigens, Protozoan - analysis; Antigens, Protozoan - immunology; Biological and medical sciences; biological development; Bradyzoites; Cell culture; Cell culture techniques; Cell Line; cell lines; chemical stress agents; chemical substances; Comparative analysis; Conversion; Cysts; Development; DEVOLOPMENT; forskolin; Fundamental and applied biological sciences. Psychology; General aspects; General aspects and techniques. Study of several systematic groups. Models; Haplorhini; Heat-Shock Proteins - analysis; Heat-Shock Proteins - immunology; Immunofluorescence; In vitro methods and tests; Infections; Invertebrates; Microscopy, Electron; Monoclonal antibodies; Neospora - growth & development; Neospora - immunology; Neospora - metabolism; Neospora - ultrastructure; Neospora caninum; neosporosis; Neurological diseases; Nitric Oxide Donors - pharmacology; Nitroprusside - pharmacology; Parasites; Parasitism; Parasitology; Protozoan Proteins - analysis; Protozoan Proteins - immunology; Sodium nitroprusside; Stress analysis; Tachyzoites; Transmission electron microscopy; Vacuoles; Germany; Parasite; Protozoa; Apicomplexa; In vitro; Stress; Neospora caninum
• ISSN: 0022-3395; 1937-2345
• DOI: 10.1645/GE-3298

Neospora caninum is an apicomplexan parasite identified as a major cause of abortion in cattle and neurological disease in various animal species. It is closely related to Toxoplasma gondii, sharing the ability to persist indefinitely in latent stage within the host as a tissue cyst containing slow-dividing bradyzoites. In this study, we compared different stress methods to induce in vitro bradyzoite conversion, using MARC-145 cells infected with Nc-Liverpool isolate. The tachyzoite-to-bradyzoite conversion rate was monitored at days 3, 5, and 7 after stress in a double-immunofluorescence assay using a monoclonal antibody against the tachyzoite antigen SAG1 (αSAG1) and a rabbit serum directed to the intracytoplasmic bradyzoite antigen BAG1 (αBAG1). Seven days of treatment with 70 μM sodium nitroprusside offered the highest bradyzoite transformation rate and the best yield of total parasitophorous vacuoles observed. In the present work, we introduce an alternative, simplified, and more advantageous method for bradyzoite production of N. caninum, using a reliable cell culture system easy to handle and with promising capacity of parasite purification.