Toxin production by Clostridium botulinum type A under various fermentation conditions

Abstract

The time of appearance and the quantity of toxin produced by the Hall strain of Clostridium botulinum type A were examined under various conditions. A 70-liter fermentor and a complex medium consisting of 2% casein hydrolysate and 1% yeast extract plus an appropriate concentration of glucose were employed. Optimal conditions for toxin production were as follows: a nitrogen overlay at a rate of 5 liters/min, an agitation rate of 50 rpm, a temperature of 35 degrees C, and an initial glucose concentration of 1.0% with the pH uncontrolled. Under these conditions, the maximum toxin concentration (6.3 x 10(5) mouse median lethal doses/ml) was attained within 24 h. Cell lysis was apparently not required to obtain maximum toxin concentrations under the fermentation conditions described.

Dependence of Clostridium botulinum gas and protease production on culture conditions

“…This report appears to contradict the belief that exogenous carbohydrates are required for the production of botulinal toxin. Examination of the literature (3, 4, 17, 18) indicates that detectable levels of toxin were produced, but carbohydrate addition caused at least a 10-fold (4) and as much as a 1,000-fold (17,18) increase in the level of toxin.…”

Comparison of the action of types A and F botulinum toxin at the rat neuromuscular junction

“…Clostridium botulinum type A strain Hall was cultivated in a fermenter system (Siegel and Metzger, 1979)9 The method used for purification of the toxin was a modification of those ,cribed previously (Duff et al, 1957;DasGupta and Boroff, 1967;Sugiyama, et al, 1977 The Langeland strain of type F was cultivated as previously described a (Yang and Sugiyama, 1975) The toxin was precipitated from the culture fluid by adjusting the pH to 4.0, extracted with 0.2M phosphate pH 6.0 and reprecipitated with amonium sulfate (Oishi and Sakaguchi, 1974). ' The a._ precipitated toxin was dissolved in 0.07 M phosphate pH 6.0, dialyzed against that buffer, and applied to a DEAN cellulose Qolumn equilibrated in the phosphate.…”

“…Toxicity was measured by mouse bioassay as previously described (Siegel and Metzger, 1979 10 days after injection, the edl nerve-muscle preparation was examined for alte-Zations in muscle mechanical properties (in situ) or electrophysiological properties (in vitro), using techniques described previously (Sellin et al, 1983a;Sellin et al, 1983b).…”

Effect of fermentation conditions on toxin production by Clostridium botulinum type B

“…High yields of toxin (6.3 x 105 mouse LD5o/ ml) have been obtained with the Hall strain of type A cultivated in a fermentor system (10). The conditions required by this strain for maximum toxin production (10) are identical to those reported here for the Bean strain of type B.…”