Factors Involved in Production of Clostridium Welchii Alpha Toxin

Abstract

Various simplified media have been described in which Clostridiura welchii grows well but produces no alpha toxin (1-3). Macfarlane and Knight (4) showed that the addition of autolyzed muscle to a medium which is satisfactory for growth enhanced the production of alpha toxin. Rogers and Knight (5) presented evidence indicating that the effect of autolyzed muscle was in part due to glucosamine. Logan, TyteU, Danielson, and Griner (6) used a pancreatic digest of beef heart as a source of "toxin factor" and furthermore demonstrated the importance of dextrin as a source of carbohydrate in obtaining maximum fields of alpha toxin with certain strains of Cl. welchii. Adams and Hendee (7) used a pancreatic digest of casein for "toxin-promoting factor" and confirmed the finding of Logan et al. that dextrin was an important factor in obtaining high yields of toxin.In the course of the present investigation it has been found that for the production of maximum yields of Cl. welchii alpha toxin, at least two factors are required in addition to dextrin and the basic growth requirements. One of these factors is present in enzymatic digests of certain proteins such as casein and gelatin. The second factor is present in extracts of pancreas and gastric mucosa. Glycerylphosphorylcholine is responsible, at least in part, for the toxin-promoting activity of pancreas and gastric mucosa.

Enterotoxin synthesis by nonsporulating cultures of Clostridium perfringens

“…Numerous studies attribute the control or regulation of protein synthesis to a specific nutrient (1,3,17,26). C. perfringens enterotoxin production may also be regulated by nutritional factors.…”

Veal Calves Produce Less Antibodies against C. Perfringens Alpha Toxin Compared to Beef Calves

“…Moreover, the current feeding regimes also provide lower amounts of milk replacer compared to the previous regimes. Contact with milk protein has been observed to decrease alpha toxin production by C. perfringens , which may influence antibody production against this toxin ([ 22 ], confirmed by unpublished data in supplementary materials ). Since the provision of solid feed did not influence the antibody dynamics of C. perfringens alpha toxin and perfringolysin, differences in quantity and quality of milk replacer used in veal calves compared to beef calves might lead to different exposure to alpha toxin, possibly explaining the difference in antibody production observed in both studies.…”

“…The pTrcHisA plasmids encoding native perfringolysin (Department of Microbiology and Immunology, College of Medicine, University of Oklahoma, USA) were transformed in chemically competent E. coli TunerTM(DE3)pLysS cells (Novagen, Darmstadt, Germany). The proteins were expressed in Terrific Broth (24% Tryptone (Oxoid, Hampshire, UK), 42% Yeast extract (BD), 4% glycerol, 0.72 M Na 2 HPO 4 , 0.16 M NaH 2 PO 4 ) and the protein expression was induced with 0.5 mM isopropyl β- d -thiogalactopyranoside (IPTG, Promega, Madison, WI, USA) [ 22 ]. The E. coli cells were lysed by sonication in a lysis buffer (pH 7.4) containing 20 mM NaPO 4 , 0.5 M NaCl, 20 mM Imidazole, 1 mg/mL lysozyme (Sigma-Aldrich, St. Louis, MO, USA) supplemented with protease-inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA).…”

Factors affecting lecithinase activity and production inClostridium welchii

“…Gordon, Turner & iDmochowski (1954) reported that lecithin inhibited the haemolytic activity of lecithinase. Although Adams, Hendee & Pappenheimer (1947) found that lecithin stimulated lecithinase production, their experiments were performed in a complex non-synthetic medium.…”