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At least 19 recordsLinked to original sources
PubMed1991

An alternative to the toxin neutralization assay in mice for the potency testing of the Clostridium tetani, Clostridium septicum, Clostridium novyi type B and Clostridium perfringens type D epsilon components of multivalent sheep vaccines.

Potency testing of veterinary vaccines containing clostridial antigens currently requires the vaccination of laboratory rabbits followed by the determination of specific antitoxin concentration in the rabbit sera by toxin neutralization test in mice. ELISAs are described as an alternative method to toxin neutralization for the determination of Clostridium tetani, Clostridium septicum, Clostridium novyi Type B and Clostridium perfringens Type D epsilon antitoxins. The assays were found to be rapid, specific and economical and showed good correlation with the toxin neutralization test.

Animals↗
PubMed1982

The effect of oxidation-reduction potential on spore germination, outgrowth, and vegetative growth of Clostridium tetani, Clostridium butyricum, and Bacillus subtilis.

Spores of Clostridium tetani germinated in liver broth with +580 mV as the starting Eh value, and those of Clostridium butyricum germinated in liver broth with an initial Eh of +400 mV regardless of the presence or absence of a liquid paraffin covering. Spores of Bacillus subtilis germinated in liver broth with--100 mV as the starting Eh value. Also, it was found that there are two ranges of starting Eh values for germination and vegetative growth of Cl. tetani, Cl. butyricum, and B. subtilis. In the first range these spores germinated and grew, but in the second range they only germinated and then died without outgrowth.

Bacillus subtilis↗
PubMed1978

The flagellar protein of Clostridium tetani.

Clostridium tetani ATCC 19406 was investigated with regard to the flagellar filaments produced by this anaerobic species. Flagellar filaments were removed from the cell bodies by hydrodynamic shear forces and purified by differential centrifugation. Exposure to acid was shown to result in disaggregation of the flagellar filaments into a preparation of flagellar protein containing 3.5% carbohydrate. The protein was judged homogeneous after examination by acrylamide gel electrophoresis in the presence of 4 M urea at several pH levels, and was shown to have a molecular weight of 35,000 daltons. Amino acid analyses indicated the absence of cysteine and tryptophan and a preponderance of acidic residues, epsilon-N-methyllysine was shown to be absent and the N-terminal amino acid was identified as alanine. Analysis of the C-terminal region indicated the sequence -Leu-Leu-Arg. These findings indicated that the obligate anaerobe C. tetani produced flagella filaments similar to previously studied filaments of aerobic and facultatively anaerobic bacteria.

Amino Acid Sequence↗
PubMed1982

High-molecular-weight hemolysin of Clostridium tetani.

Clostridium tetani excretes hemolysins of two size classes, a high-molecular-weight hemolysin (HMH), which was eluted near void volume of a Sepharose 6B column, and conventional tetanolysin (molecular weight, approximately 50,000). The total hemolysin activity in the culture supernatant increased sharply with growth of bacteria and remained at a high level during autolysis. The content of HMH, however, decreased from 41% at 4 h of culture to 0.4% at the early stage of autolysis. The cell bodies also exhibited hemolytic activity, 70% of which could be solubilized and separated into HMH and the 50,000 Mr tetanolysin as extracellular hemolysins. The activity ratio of HMH to the total solubilized hemolysins was 0.45, on the average, at 6 h of culture but was 0.23 at the middle of logarithmic growth. Partially purified HMH from both sources appeared as broken pieces of cytoplasmic membranes under an electron microscope. The ratio of proteins to phospholipids in HMH was found to 3.26, a value similar to that in cell membrane. The total cell hemolytic activity decreased by 90 or 75% upon addition of chloramphenicol or anti-tetanolysin serum, respectively, into a 6-h-old culture of bacteria. It is suggested that HMH is a complex of tetanolysin with a membrane fragment and releases the conventional tetanolysin during bacterial culture.

Bacterial Toxins↗
PubMed2005

BotR/A and TetR are alternative RNA polymerase sigma factors controlling the expression of the neurotoxin and associated protein genes in Clostridium botulinum type A and Clostridium tetani.

Clostridium botulinum and Clostridium tetani, respectively, produce potent toxins, botulinum neurotoxin (BoNT) and tetanus neurotoxin (TeTx), which are responsible for severe diseases, botulism and tetanus. Neurotoxin synthesis is a regulated process in Clostridium. The genes botR/A in C. botulinum A and tetR in C. tetani positively regulate expression of BoNT/A and associated non-toxic proteins (ANTPs), as well as TeTx respectively. The botR/A gene lies in close vicinity of the two operons which contain bont/A and antps genes in C. botulinum A, and tetR immediately precedes the tetX gene in C. tetani. We show that BotR/A and TetR function as specific alternative sigma factors rather than positive regulators based on the following results: (i) BotR/A and TetR associated with target DNAs only in the presence of the RNA polymerase core enzyme (Core), (ii) BotR/A and TetR directly bound with the core enzyme, (iii) BotR/A-Core recognized -35 and -10 regions of ntnh-bont/A promoter and (iv) BotR/A and TetR triggered in vitro transcription from the target promoters. In C. botulinum A, bont/A and antps genes are transcribed as bi- and tricistronic operons controlled by BotR/A. BotR/A and TetR are seemingly related to a new subgroup of the sigma70 family that includes TcdR and UviA, which, respectively, regulate production of toxins A and B in C. difficile and bacteriocin in C. perfringens. Sequences of -35 region are highly conserved in the promoter of target toxin genes in C. botulinum, C. tetani, C. difficile and C. perfringens. Overall, a common regulation mechanism probably controls toxin gene expression in these four toxigenic clostridial species.

Bacterial Proteins↗
PubMed1979

Taxonomy of Clostridium tetani and related species.

Clostridium tetani and its related species C. tetanomorphum, C. cochlearium and C. lentoputrescens were examined for DNA-DNA homology and biochemical properties. Two distinctly different groups were included under the name of C. tetanomorphum: one was identical with C. cochlearium and the name C. tetanomorphum was applied to the other group with some amendment of biochemical properties. Comparison of the type strain of C. lentoputrescens with wild strains obtained from horse faeces indicated that the name C. lentoputrescens should be abolished as a later synonym of C. cochlearium. Liquefaction of gelatin and spore shape, which have been used as the important criteria for differentiation of C. tetani-related species, were genetically insignificant.

Base Sequence↗
PubMed1986

Density and distribution of Clostridium tetani in the soil.

Clostridium tetani or its toxin was recovered from a minute amount of soil samples collected on the surface of the ground of various places. C. tetani was recovered from 10% and tetanus toxin alone from another 1% of soil samples employing 1 mg of the soil. C. tetani or its toxin alone was recovered, employing 1 mg or more of soil, from the wet shores of ponds and rivers, fields and rice fields (85%), the yards of farmers and non-farmers (53%), school and hospital grounds (30%) and on the roadside (20%). Two tetanus patients were injured in their yards where 1 mg soil samples yielded C. tetani. 12 of 13 soil samples collected at the yard of one of these patients at 3 different times yielded C. tetani. C. tetani did not remain at the same place in the same quantity. It was isolated more readily from the soil samples collected on the surface of than deep in the ground.

Clostridium tetani↗
PubMed1987

Colonization of the intestinal tract of mice with Clostridium tetani.

Colonization of intragastrically inoculated Clostridium tetani in intestinal tract of mice was investigated by cultivation of daily stool samples and quantification of the bacilli over a period of 3 weeks. We found that most of the inoculum was excreted in the stool during the first 24 hr. The amount of C. tetani recovered on the second day after inoculation was very small. C. tetani apparently thrived in the intestinal tract when more than 6 log colony forming units (CFU) of tetanus spores had been inoculated. Otherwise, C. tetani colonized poorly in intestinal tract. Tetanus toxin was not detected in the stool nor in the contents of the small and large intestines 24 hr after 6.23 log CFU of tetanus spores had been inoculated. We concluded that the intestinal cavity of mouse is not a favorable environment for colonization of C. tetani under natural conditions.

Animals↗
PubMed1988

Some factors affecting isolation of Clostridium tetani from human and animal stools.

Clostridium tetani was isolated from human and animal stools at the following rates [95% confidence interval (CI)]: Human, 0% (1.5-0); horse, 1% (5-0); cow in cowshed, 4% (10-1); cow in pasture, 8.3% (17-1), calf in pasture, 0% (7-0); dog, 2% (11-0) and sheep in pasture, 25% (44-14). Quantification of C. tetani in 16 animal stools positive for the bacillus was impossible in most cases, as the number of tetanus bacilli present was not large enough for this purpose. Contaminating anaerobic and facultative anaerobic bacteria in human and animal stools, i.e., C. perfringens and Streptococcus sp., Group G, inhibited isolation of C. tetani from these materials, particularly at the step of isolation employing its swarming character.

Adult↗
PubMed1968

Continuous production of Clostridium tetani toxin.

The continuous production of Clostridium tetani toxin has been carried out in a 1-liter stirred culture vessel for as long as 65 days. Toxin production of approximately 120 flocculating units per ml was maintained with a dilution rate of 0.125 hr(-1), a temperature of 34 C, a pH of 7.4, and the addition to the medium of 0.1 g of potassium chloride per liter. The average minimal lethal intraperitoneal dose of the toxin in mice was approximately 10(6) per ml.

Bacteriological Techniques↗
PubMed1965

ISOLATION OF CLOSTRIDIUM TETANI FROM SOIL.

Sanada, Ichiro (Kanazawa University, Kanazawa, Japan), and Shoki Nishida. Isolation of Clostridium tetani from soil. J. Bacteriol. 89:626-629. 1965.-The higher the temperatures applied to soil specimens, the weaker the toxigenicity of Clostridium tetani strains isolated from them. The glucose- and maltose-fermenting ability of these isolates was inversely proportional to their toxigenicity. The biological properties of atoxic strains were indistinguishable from those of C. tetanomorphum. Since a considerable number of toxic strains fermented glucose and maltose, these criteria are of doubtful value for differentiating C. tetani from C. tetanomorphum.

Antitoxins↗