Arginine Promotes Toxin Formation in Cheddar Cheese by Clostridium botulinum

Abstract

The production of botulinal toxin by a mixture of spores of Clostridium botulinum types A and B was evaluated in Cheddar cheese supplemented with L-arginine (1% wt/wt) and containing one of three levels of sodium chloride (0, 0.9, or 1.8%). Botulinal toxin was formed in cheeses containing an increased level of L-arginine (1%) and reduced levels of sodium chloride (0 or 0.9%). No toxin was formed in Cheddar with arginine and 1.8% salt or in any of the cheeses not supplemented with arginine. The pH increased from 5.05–5.2 to 5.7–6.0 in the cheeses with increased arginine, but the pH change alone did not permit growth of C. botulinum. Metabolism of arginine may also have promoted the synthesis of compatible metabolites for salt resistance. The results indicate that an important factor supporting growth of C. botulinum in cheese is the availability of L-arginine.

Quantification of Clostridium botulinum Toxin Gene Expression by Competitive Reverse Transcription-PCR

“…Expression of the C. botulinum E VH toxin gene was detected by the mouse bioassay in the early stationary phase when the organism was grown in BHI broth and type E broth. Extraction and the assay for botulinal toxin were performed as described by Malizo et al (13). Cells from 200-ml broth cultures were concentrated by centrifugation at 7,500 ϫ g for 30 min at 4°C.…”

Clostridium botulinumandClostridium tetani

“…This grouping is also supported by metabolic and structural properties including nutritional requirements (Belokopytov et al 1982;Kindler et al 1956;Mager et al 1954;Malizio et al 1993;Patterson-Curtis and Johnson 1992;Whitmer and Johnson 1988), resistance to salt and acidity and other environmental and food components (Anniballi et al 2002;Glass and Johnson 2001;Hammer and Johnson 1988;Hutchinson 1992;Odlaug and Pflug 1978;Ohye and Scott 1957;Raatjes and Smelt 1979;Lynt et al 1982;Sugiyama and Sofos 1988;Townsend et al 1954), spore heat resistance and germination properties (Broussolle et al 2002;Ito et al 1968;Lund and Peck 2000;Lynt et al 1975;Scott and Bernard 1982;Setlow and Johnson 2001), tolerance to air (Meyer 1924;Whiting and Naftulin 1992), minimum growth temperature (Eklund et al 1976;Smith and Sugiyama 1988), endproduct formation (Mead 1971;Moore et al 1966;Moss et al 1970Moss et al , 1980Oguma et al 1986;Smith and Sugiyama 1988), and toxin regulation. The four groups also have distinctive surface antigen relationships (Batty and Walker 1966;Hatheway 1983, 1984a, b;Lynt et al 1967;Solomon et al 1969Solomon et al , 1971…”

“…Arginine is the organic nutrient required in highest concentrations by group I C. botulinum and strongly represses toxin and protease formation in C. botulinum types A and B (Patterson-Curtis and Johnson 1989. Arginine promotes toxin formation by C. botulinum in moderately acidic foods (Malizio et al 1993;Patterson-Curtis and Johnson 1992) and probably also protects the bacterium against environmental stresses including acidity and high osmolarity (Patterson-Curtis and .…”