• Journal of Food Hygiene and Safety
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• v.8 no.1
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• pp.55-61
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• 1993

Soil samples and the intestinal contents of arthropods, mollusca, pisces, aves, and mammals were examined for the presence of Clostridium botulinum. Demonstration of Clostridium botulimun was accomplished by identifying its toxin in liquid cultures inoculated with soil or material from the alimentary tract of tested animals with toxin neutralization tests in addition to morphological, cultural and biochemical tests. Incidences of Clostridium botulinum in tested samples were 5.0% in soil, 6.7% in mammal and 8.7% in fish, respectively. All of the positive cultures were identified as Clostridium botulinum type E and any other type was not demonstrated throughout the survey. Canned foods and solid ham/sausage mixture formulated as can with distilled water were inoculated with Clostridium botulinum type E and checked for toxin production by using the mouse bioassay. Clostridium botulinum type E toxin was produced as a large quantity in canned foods of fish, shell, meat and ham and, however, no significant toxin was detected in sausages and fruit samples.

• Korean Journal of Veterinary Service
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• v.22 no.4
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• pp.349-356
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• 1999

Type C botulism was broken out in cattle at Pochun country, Kyonggi in Korea. The main cause was on the feed of fermented food contaminated with Clostridium botulinum. It was supposed to be contaminated with C botulinum during producing the fermented food because of either using contaminated food residues or adding contaminated additive materials like sausage. One hundred and twenty out of 642 cattle in 11 farms showed typical botulism symptoms and 95 among them died. The incubation period was from 2 to 14 days and the average incubation period was 8 days. The mortality in this case was 18.3%.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.499-505
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• 2006

Clostridium botulinum spores are widely distributed in nature. Type A and proteolytic type B bacteria produce heat-resistant spores that are primarily involved in most of the food-borne botulism outbreaks associated with low-acid canned foods. Food-borne botulism results from the consumption of food in which C. botulinum has grown and produced neurotoxin. Growth and toxin production of type A and proteolytic type B in canned foods can be prevented by the use of thermal sterilization alone or in combination with salt and nitrite. The hazardousness of C. botulinum in low-acid canned foods can also be reduced by preventing post-process contamination and introducing hazard analysis and critical control point (HACCP) practices during production. Effectiveness of non-thermal technologies such as high pressure processing with elevated process temperatures on inactivation of spores of C. botulinum will be discussed.

• Toxicological Research
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• v.13 no.4
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• pp.417-421
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• 1997

Synaptobrevin is a kind of vesicle associated membrane proteins (VAMPs) which plays a secretary role in the neuronal synapse and was recently known as the biochemical target of botulinum neurotoxin type B. The structural gene of the synaptobrevin was cloned from mouse brain using RT-PCR technique and was seqrtenced. The deduced amino acid sequence showed that the synaptobrevin protein from mouse brain is exactly the same with that of the rat brain in the amino acid level. The synaptobrevin gene was subcloned into pET3a vector and expressed in E. coli. The molecular weight of the recombinant protein was 19 kDa as expected. Moreover, when the recombinant synaptobrevin protein was incubated with the native neurotoxin of Clostridium botulinum type B, it was cleaved by the toxin in a time dependent manner. This implies that the recombinant synaptobrevin protein and the native toxin are reacted in the same way as the native synaptobrevin did in the neuronal cells.

• Korean Journal of Microbiology
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• v.20 no.4
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• pp.183-188
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• 1982

The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163, 000. Subunits of 106, 000 and 56, 000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

• Microbiology and Biotechnology Letters
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• v.10 no.3
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• pp.205-209
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• 1982

The enzyme-linked immunosorbent assay using the so-called "double-sandwich"technique was applied to determine Clostridium botulinum type F toxin. Polystyrene tubes were coated with horse anti-type F toxin serum and then toxin sample was added. The tubes were subsequently treated with rabbit anti-type F toxin IgG and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate. By this technique, about 10 mouse intraperitoneal 50% lethal doses (ip LD/50/) of type F toxin could be detected. Low back-ground reading was achieved with the use of phosphate-buffered saline containing 0.05% Tween 20 and 1% bovine serum albumin as diluents of rabbit IgG and conjugate. Addition of EDTA in the diluents of toxin increased ELISA extinction value significantly. No cross-reaction was observed with botulinum type A and B toxin, but type E toxin gave sleight cross-reaction.

• Journal of Microbiology
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• v.41 no.2
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• pp.161-164
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• 2003

Recombinant light chain of Clostridium botulinum neurotoxin type B stimulated transglutaminase activity in a dose dependent manner, Compared to native toxin, recombinant light chain showed av greater stimulatory effect on transglutaminase activity. Zn-chelating agents, inhibiting the proteolytic activity of the clostridial toxins, did not interfere with this stimulation. These results suggest that the light chain plays a major stimulatory role, which is not due to its metallopeptidase activity, but is possibly due to specific interaction with transglutaminase. More importantly, this report provides a new insight into the intracellular action of C. botulinum neurotoxins.

• Preventive Nutrition and Food Science
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• v.12 no.4
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• pp.259-266
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• 2007

It has been proposed that the mode of action of nisin against vegetative cells and spores of Clostridium botulinum is different. However, clear explanation is not available. Therefore, nisin action against vegetative cells and spores of C. botulinum was investigated in this study. Nisin was added at various stages of spore-to-vegetative cell transition and changes to sensitivity to the bacteriocin were observed. Different nisin preparation (Nisaplin or pure nisin) was compared for their activity against different stages of spore transformation of C. botulinum ATCC 25763. Germination was measured by determining loss of heat resistance and observing phase darkening of spores under phase-contrast microscope. Nisin acted bactericidally against vegetative cells, but acted sporostatically against spores of C. botulinum under the same concentration. This bactericidal and sporostatic action of nisin was dependent on the concentration of nisin used. Presence of nisin during spore activation by heat increased subsequent phase darkening and germination rates. However, nisin inhibited the germination and the outgrowth, when it was added after heat activation stage. Findings from this study suggest that the time of addition of nisin is very important for the effective control of spores during the heating process of foods. In addition, it may be possible to apply nisin at the stage of processing that coincides with the most sensitive stage of spore transformation.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.287-291
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• 2003

Botulinum neurotoxin type A (BONT/A) is an extremely potent toxin, which is produced by Clostridium botulinum. The light chain of this protein (BONT/A LC), which is known as a zinc endopeptidase, cleaves SNAP-25 involved in the exocytosis process. In this work, the expression of recombinant BoNT/A LC in E. coli is described. The BONT/A LC gene of C. botulinum contains a high frequency of the arginine AGA and isoleucine ATA codons that are rarely used in genes of E. coli, hampering the translation of recombinant protein. The argD and ilex tRNA genes were cloned into pACYC184 vector, resulting in pAAD131X plasmid. The translational stress of the toxin gene related to codon bias was reversed by fupplernentation of the AGA arginyl tRNA of T4 phage and AUA isoleucyl tRNA of E. coli. This system may be applicable for the expression of a variety of AT-rich heterologous genes in E. coli.

• Korean Journal of Veterinary Research
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• v.48 no.2
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• pp.161-165
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• 2008

Many neurotoxigenic clostridia are found in soil. Among animals, birds are especially susceptible to botulism, perhaps because they feed on insects, invertebrate carcasses, and decayed feeds contaminated with spores of Clostridium (C.) botulinum. C. botulinum type C is mainly involved in avian botulism. In the summer of 2005, death of a mute swan (cygnus olor) living in the pond of large bird cage was found in Seoul Grand Park Zoo. The birds presented presumptive clinical signs of botulism, such as ruffled hackle feathers, abnormal posture of the head, weakness, and flaccid paralysis. At that time, pond water in the breeding facilities was drained for 7 days, but there were still remained water containing sediment of feed and feces. Therefore, botulism was suspected and an experimentation were made to detect C. botulinum in the dead mute swan. Gross post-mortem findings of a mute swan showed jelly-like hemorrhagic contents in the intestine, sands and vegetations in the stomach. C. botulinum was isolated from the liver, small intestine and large intestine samples. Botulism was also confirmed by mouse inoculation test with the organ samples. With PCR, a gene encoding C. botulinum type C toxin was detected for the several organs of the mute swan died. These results suggested that death of mute swan was caused by C. botulinum type C.