• 한국식품위생안전성학회지
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• 제8권1호
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• pp.55-61
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• 1993

토양을 비롯하여 어류, 패류, 조류, 포유류의 소화기관으로부터 Clostridium botulinum 분리를 시도하였다. 총 158개 분리원을 screening한 결과 10개 시료로부터 Clostridium botulinum 분포 가능성을 확인하였으며 6개 시료로부터 Clostridium botulinum을 순수 분리하여 3.8% 분리율을 나타내었다. 분리 균주의 형태적 특징, 배양상의 특성 및 생화학적 특성 등을 표준 균주의 특성과 비교하고 항혈청에 의한 중화시험을 실시하여 분리균주를 동정하였다. Egg york agar에서의 opalescence 생성, 탄수화물 이용성, Egg york GAM 배지상에서의 pearly layer 생성 등으로부터 Clostridium botulinum으로 동정할 수 있었으며 trypsin에 의한 toxicity 활성화, type E 항혈청에 의한 opalescence 생성억제 및 mouse 방어효과가 인정되어 type E 로 동정하였다. 국내에서 시판되고 있는 수 종의 식품을 대상으로 Clostridium botulinum 의 toxin 생성능을 비교하였던 바 식품의 종류, 사용균주에 따라 toxin 생성량에 현저한 차이가 있었다. 분리균주 type E 의 경우 어패류통조림, ham 식품에서 많은 양의 toxin 이 생성되었으며 sausage, 과일통조림 식품에서는 비교적 적었다. 그러나 type A 의 경우에는 어패류, ham , sausage 식품에서 상당량의 toxin 이 생성되었으며 과일통조림에서도 비교적 맣은 양의 toxin이 생성되었다 .

• 한국동물위생학회지
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• 제22권4호
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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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• 제15권4호
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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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• 제13권4호
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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.

• 미생물학회지
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• 제20권4호
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• pp.183-188
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• 1982

Clostidium botulinum type B가 생성하는 독소를 정제할 수 있는 방법을 연구하였다. 정제과정은 독소를 ammonium sulfate로 배양액에서 침전시켜 추출한후 Polymin P를 처리하여 핵산 및 기타 단백질을 최대한 제거한 후 Sephaex G-I00에서 gel fiItration을 시키고 DEAE-Sephadex로 이온교환 크로마토그래피를 시켰다. 이러한 과정으로 정제된 독소의 회수율은 17%였으며 SDS-polyacrylamide gel electrophoresis 결과 하나의 선을 나타내 동질성을 증명하였다. 정제된 독소의 분자량은 163,000이였으며 $\beta$-mercaptoethanol을 사용하여 환원시킨 결과 분자량 106,000과 56,000의 하위 단위체로 분리되었다.

• 한국미생물·생명공학회지
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• 제10권3호
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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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• 제41권2호
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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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• 제12권4호
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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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• 제13권2호
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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.

• 대한수의학회지
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• 제48권2호
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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.