Kinetics study of immunological response to Clostridium botulinum toxin.
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A double-antibody enzyme-linked immunosorbent assay (ELISA) for detection of humoral antibody to type A botulinal toxin was developed. This assay was used to study the kinetics of antibody response of a volunteer to botulinal toxoid. The circulating type A antitoxin was first detected by the ELISA 2 weeks after the first booster injection of the toxoid. The antibody titer stayed level until the second booster at 12 weeks. The titer then continued to rise throughout the remaining study period. The neutralizing antibody to type A toxin was detected by mouse assay 15 weeks after detection of antitoxin by the ELISA. (+info)
Purification and characterization of neurotoxin produced by Clostridium botulinum type C 6813.
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The toxin produced by Clostridium botulinum type C 6813 (C-6813) was purified 1,009-fold from the culture supernatant in an overall yield of 30%. The specific toxicity was 1.1 X 10(7) mouse minimum lethal doses per mg of protein. The toxin had a molecular weight of 144,000, composed of the light and heavy chains with molecular weights of 52,000 and 92,000, respectively, linked by one or two disulfide bond(s). The purified C-6813 toxin heavy and light chains reacted strongly with anti-type D heavy chain immunoglobulin G and anti-type C1 light chain immunoglobulin G, respectively. The amino acid compositions of C-6813 toxin heavy and light chains were more similar to those of type D heavy chain and type C1 light chain than to those of type C1 heavy chain and type D light chain, respectively. These results suggest that in the toxin produced by the type C strain at least two subtypes exist. (+info)
Development of antitoxin with each of two complementary fragments of Clostridium botulinum type B derivative toxin.
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Two fragments with molecular weights of 111,000 (fragment I) and 59,000 (fragment II) were separated from each other by gel filtration of dithiothreitol and urea-treated, trypsinized derivative toxin (molecular weight, 170,000) of the proteolytic Okra strain of Clostridium botulinum type B on a column of Sephadex G-200 (superfine) with a buffer containing dithiothreitol and urea. Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule. Both fragments were immunogenic, and both anti-fragments neutralized type B toxin. The neutralizing activities of both anti-fragment I and anti-fragment II were, however, lower than that of the anti-derivative toxin, suggesting that the molecular integrity of derivative toxin is essential for sufficient production of the neutralizing antibody. The immunological difference found between type B toxin from a proteolytic strain and that from a nonproteolytic strain was ascribed to the antigenic difference of fragment I. (+info)
Activation of a toxic component of Clostridium botulinum types C and D by trypsin.
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When the Stockholm and 468C strains of type C and the 1873 strain of type D Clostridium botulinum are "cured" of their prophages, they simultaneously discontinue the production of their dominant toxins (C(1) and D), but they continue to produce a second antigenically monospecific toxin (C(2)). These "cured" strains of types C and D therefore become indistinguishable with respect to the toxin produced. Fifteen type C cultures received from other laboratories discontinued to produce the dominant toxin when subcultured in broth. The C(2) toxin, however, was produced by eight of these cultures. The C(2) toxin is produced by these cultures as a protoxin that requires treatment with trypsin before its toxicity can be demonstrated. Of the 21 type C cultures that produce the C(1) toxin, 20 were shown to produce the C(2) toxin. The filtrates of 14 of these cultures required trypsin treatment before the C(2) toxicity could be demonstrated. Low levels of toxicity could be demonstrated in the six remaining culture fluids without trypsin; toxicity, however, was increased with trypsin. (+info)
Clostridium botulinum type F: isolation from venison jerky.
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A Clostridium botulinum type F was isolated from the venison jerky responsible for the only type F botulism outbreak reported in the United States. The isolate differed from the prototype Langeland type F strain in being nonproteolytic. (+info)
Detection of type E botulinal toxin in cultures by fluorescent-antibody microscopy.
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Vegetative cells of toxigenic Clostridium botulinum type E cultures were stained with fluorescent antitoxin prepared against purified toxin. The staining seems to be specific. (+info)
Human botulism in Canada (1919-1973).
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Since 1919, in Canada, 62 authenticated outbreaks of human botulism have affected 181 persons, with 83 deaths, a fatality rate of 46%. Among these, 41 outbreaks were bacteriologically determined (31 in one laboratory) as six type A, four type B, one both A and B, and 30 type E. About two thirds of the total outbreaks, cases and deaths involved Eskimos and Pacific coast Indians consuming raw marine mammal products and salmon eggs, respectively. Other parts of Canada recorded seven occurrences due to miscellaneous vehicles, three being type B. Since January 1961 there have been 38 outbreaks, involving 94 cases with 33 deaths. These include 18 outbreaks among Eskimos, affecting 51 persons (of whom 24 died) in Labrador, southern Baffin Island, northern Quebec, and the Mackenzie area. Also, putrid salmon eggs caused 15 outbreaks among Pacific coast Indians, totalling 35 cases, of whom only six died, the low fatality rate reflecting the introduction of type E botulinus antitoxin during 1961. (+info)
Evaluation of type A botulinal toxin assays that use antitoxin to crystalline toxin.
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The type A botulinal toxin assay by the reverse passive hemagglutination procedure which uses antitoxin to crystalline toxin was examined for specificity. The analysis was based on the fact that crystalline type A toxin is a complex of neurotoxic protein (Aalpha) and a nontoxic protein (Abeta). By using these components, obtained in essentially pure forms, it was shown that the antitoxin to crystalline toxin has a significantly higher titer to Abeta than to Aalpha. When Formalin-treated red blood cells were sensitized with this antitoxin, the antibodies coupled to the cells were, for practical results, only anti-Abeta. When the suspension is reacted with dilutions of type A toxic solutions, the limiting dilutions are determined by Abeta and not by the neurotoxin, which should be the determinant if the assay is to measure toxicity. These observations may be pertinent to the development of serological assays for other botulinal toxin types. (+info)