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    CLOSTRIDIUM | Detection of Neurotoxins of Clostridium botulinum
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    An outbreak of human botulism was due to consumption of ham containing botulinum neurotoxins B and E. A Clostridium botulinum type E strain isolated from ham was assigned to a new subtype (E12) based on bont/E gene sequencing and belongs to a new multilocus sequence subtype, as analyzed by whole-genome sequencing.
    Botulism
    Clostridium botulinum
    Multilocus sequence typing
    Food poisoning
    Food microbiology
    Strain (injury)
    Citations (28)
    Heat treatment is indispensable in fish canning to provide an acceptable shelf life. Its optimisation reduces the risk of the presence of Clostridium botulinum spores, which could potentially cause botulism cases. This study evaluated canned fish samples for botulism neurotoxin (BoNT)-producing clostridia contamination and can bulging through microbiological contaminant growth. A new analytical approach was developed for detection of such clostridia and phenotypically similar species. A total of 70 canned fish samples suspected of exhibiting bulging features were analysed. Culture methods were used to detect clostridia. The isolates obtained were evaluated on the basis of the exhibited phenotypic characteristics. Also, PCRs were used for the detection of genes determining BoNT production (non-toxic non-haemagglutinin (ntnh) genes) and the amplification of conservative 16S rDNA genes, which were Sanger sequenced. The obtained sequences were analysed using the Basic Local Alignment Search Tool. Clostridium genus species were isolated from 17 (24%) bulging and organoleptically changed samples. No ntnh genes were present in these isolates; however, sequencing confirmed the presence of C. sporogenes, a species with close affinity to C. botulinum. To eliminate the threat of foodborne botulism, laboratory diagnostic techniques must detect species of the Clostridium genus and elucidate their ability to produce BoNTs. Although Clostridium botulinum is the most common cause of botulism, the possibility may not be ignored that non-pathogenic Clostridium species may acquire botulinum toxigenicity. The similarity between the isolated strains of C. sporogenes and C. botulinum should be incorporated in the optimisation of heat treatment to guarantee a sterilised, microbiologically safe product.
    Clostridia
    Botulinum neurotoxin
    Neurotoxin
    Clostridium botulinum
    Citations (1)
    Three strains of OS variants of Clostridium botulinum type E (Dolman, 1957) have been studied using additional biochemical tests and fluorescent antisera. The relationship of these organisms, and other nontoxic clostridia resembling Cl. botulinum , to typical toxic strains is discussed.
    Clostridia
    Clostridium botulinum
    Case records of 31 herd outbreaks confirmed by the Pennsylvania Botulism Diagnostic Laboratory [PBDL] over the past 13 years (1984-1997) were reviewed. Only cases with positive isolation of the Clostridia botulinum toxin or culture of Clostridia botulinum spores capable of toxin production were included in our review.
    Botulism
    Clostridia
    Clostridium botulinum
    Isolation
    Citations (1)
    SUMMARY Botulism is a potentially lethal paralytic disease caused by botulinum neurotoxin. Human pathogenic neurotoxins of types A, B, E, and F are produced by a diverse group of anaerobic spore-forming bacteria, including Clostridium botulinum groups I and II, Clostridium butyricum, and Clostridium baratii. The routine laboratory diagnostics of botulism is based on the detection of botulinum neurotoxin in the patient. Detection of toxin-producing clostridia in the patient and/or the vehicle confirms the diagnosis. The neurotoxin detection is based on the mouse lethality assay. Sensitive and rapid in vitro assays have been developed, but they have not yet been appropriately validated on clinical and food matrices. Culture methods for C. botulinum are poorly developed, and efficient isolation and identification tools are lacking. Molecular techniques targeted to the neurotoxin genes are ideal for the detection and identification of C. botulinum, but they do not detect biologically active neurotoxin and should not be used alone. Apart from rapid diagnosis, the laboratory diagnostics of botulism should aim at increasing our understanding of the epidemiology and prevention of the disease. Therefore, the toxin-producing organisms should be routinely isolated from the patient and the vehicle. The physiological group and genetic traits of the isolates should be determined.
    Botulism
    Clostridium botulinum
    Neurotoxin
    Botulinum neurotoxin
    Clostridia
    Clostridium butyricum
    Genus and Species Definitions Historical Background of the Pathogenic Clostridia Milestones in the Understanding of Botulism and Tetanus Classification of Neurotoxigenic Clostridia Habitats of Pathogenic Clostridia Physical and Metabolic Properties of Pathogenic Clostridia including C. botulinum and C. tetani Growth Properties of C. botulinum and C. tetani Physiology of C. botulinum and C. tetani Descriptions of the Species C. botulinum, C. tetani, and other Neurotoxigenic Clostridia Botulinum Neurotoxins Detection of Botulinum and Tetanus Neurotoxins Genomics of Clostridium tetani and Clostridium botulinum Susceptibility of Neurotoxigenic Clostridia to Chemical and Physical Agents Epidemiology and Clinical Aspects Use of Botulinum Toxin in Medicine Safety Precautions for Working with C. botulinum, C. tetani, and their Neurotoxins Acknowledgments
    Clostridia
    Clostridium tetani
    Clostridium botulinum
    Botulism
    Clostridiales
    ABSTRACT Botulinum neurotoxin (BoNT), the most toxic substance known, is produced by the spore-forming bacterium Clostridium botulinum and, in rare cases, also by some strains of Clostridium butyricum and Clostridium baratii . The standard procedure for definitive detection of BoNT-producing clostridia is a culture method combined with neurotoxin detection using a standard mouse bioassay (SMB). The SMB is highly sensitive and specific, but it is expensive and time-consuming and there are ethical concerns due to use of laboratory animals. PCR provides a rapid alternative for initial screening for BoNT-producing clostridia. In this study, a previously described multiplex PCR assay was modified to detect all type A, B, E, and F neurotoxin genes in isolated strains and in clinical, food, environmental samples. This assay includes an internal amplification control. The effectiveness of the multiplex PCR method for detecting clostridia possessing type A, B, E, and F neurotoxin genes was evaluated by direct comparison with the SMB. This method showed 100% inclusivity and 100% exclusivity when 182 BoNT-producing clostridia and 21 other bacterial strains were used. The relative accuracy of the multiplex PCR and SMB was evaluated using 532 clinical, food, and environmental samples and was estimated to be 99.2%. The multiplex PCR was also used to investigate 110 freshly collected food and environmental samples, and 4 of the 110 samples (3.6%) were positive for BoNT-encoding genes.
    Clostridia
    Clostridium botulinum
    Neurotoxin
    Multiplex
    Botulism
    Citations (95)