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Title: Progress in culture and subculture of spheroplasts and fastidious acid-fast bacilli isolated from intestinal tissues.
Title Abreviation: J Clin Microbiol Date of Pub: 1988 Aug
Author: Markesich DC; Graham DY; Yoshimura HH;
Issue/Part/Supplement: 8 Volume Issue: 26 Pagination: 1600-3
MESH Headings: Comparative Study; Crohn Disease (*MI); Culture Media (*); Human; Hydrogen-Ion Concentration; Intestines (*MI); Mycobacterium (*GD); Paratuberculosis (MI); Spheroplasts (IP/*PH); Support, Non-U.S. Gov't; Support, U.S. Gov't, Non-P.H.S.; -RN-;
Journal Title Code: HSH Publication Type: JOURNAL ARTICLE
Date of Entry: 881123N Entry Month: 8901
Country: UNITED STATES Index Priority: 2
Language: Eng Unique Identifier: 89008819
Unique Identifier: 89008819 ISSN: 0095-1137
Abstract: The efficiency of culture media was compared for the culture and subculture of very slowly growing acid-fast bacilli and spheroplast forms obtained from intestinal tissues of patients with Crohn's disease and ulcerative colitis and from controls without inflammatory bowel disease. Media were developed by modifying a nutrient broth medium based on veal infusion broth and yeast extract. We evaluated the effects of pH and the addition of Tween 80, Dubo oleic albumin complex, an extract from intestinal tissue from a patient with Crohn's disease, horse serum, sucrose, magnesium sulfate, ferrous ammonium sulfate, and sodium citrate. All media contained mycobactin J (2 micrograms/ml). We developed a medium (MG3) which was highly successful in promoting the growth of very fastidious organisms and promoted reversion of spheroplasts to acid-fast rods. MG3 contained veal infusion broth, 1% yeast extract, 10% horse serum, 0.3 M sucrose, 0.2% MgSO4, 0.1% ferrous ammonium sulfate, 0.1% sodium citrate, and 2 mg of mycobactin J per liter. We were able to obtain quantities of organisms sufficient for examination of the organisms by molecular techniques. Successful cultivation of all isolates and reversion of spheroplasts to acid-fast forms encourage further studies of the possibility of a complex association of mycobacteria and Crohn's disease.
Abstract By: Author
Address: Inflammatory Bowel Disease Laboratory, Veterans Administration Medical Center, Houston, Texas 77030.