TOXIN PRODUCTION BY CLOSTRIDIUM TETANI IN BIPHASIC LIQUID CULTURE,
Abstract
An exploratory study is described on the use of a biphasic culture technique for production of toxin by C. tetani. The biphasic system used was a combination of dextran and polyethylene glycol (PEG) in the presence of a protein-free medium. The system supported food growth and gave a favorable phase ratio (top/bottom) of 15:1. The majority of the bacterial cells were in the bottom dextran phase, while the greater part of the toxin was in the upper PEG-medium phase. Toxin yields from a typical experiment were 10 to the 9th power mouse intraperitoneal minimum lethal doses per milliliter of upper phase filtrate. The biphasic culture system was also adaptable to continuous culture, with approximately similar toxin levels found in the upper phase. The appearance of the clostridial cells in older cultures suggested the possibility of protoplast formation, perhaps stabilized by the polymers present. In vitro toxin titration (Lf) was inhibited by the presence of the polymers. Bacillus anthracis also grew well in the biphasic system and elaborated protective antigen, primarily into the upper phase. (Author)
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 1965
- Accession Number
- AD0461500
Entities
People
- Carl-goran Heden
- Milton Puziss
Organizations
- United States Army Biological Warfare Laboratories