Characterization of  Clostridium novyi  isolated from a sow in a sudden death case in Korea

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Abstract

Background

Multifocal spherical nonstaining cavities and gram-positive, rod-shaped, and endospore-forming bacteria were found in the liver of a sow that died suddenly. Clostridium novyi type B was identified and isolated from the sudden death case, and the isolate was characterized by molecular analyses and bioassays in the current study.

Results

C. novyi was isolated from the liver of a sow that died suddenly and was confirmed as C. novyi type B by differential PCR. The C. novyi isolate fermented glucose and maltose and demonstrated lecithinase activity, and the cell-free culture supernatant of the C. novyi isolate exhibited cytotoxicity toward Vero cells, demonstrating that the isolate produces toxins. In addition, whole-genome sequencing of the C. novyi isolate was performed, and the complete sequences of the chromosome (2.29 Mbp) and two plasmids (134 and 68 kbp) were identified for the first time. Based on genome annotation, 7 genes were identified as glycosyltransferases, which are known as alpha toxins; 23 genes were found to be related to sporulation; 12 genes were found to be related to germination; and 20 genes were found to be related to chemotaxis.

Conclusion

C. novyi type B was isolated from a sow in a sudden death case and confirmed by biochemical and molecular characterization. Various virulence-associated genes were identified for the first time based on whole-genome sequencing.

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Most cited references 27

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Glucosylation of Rho proteins by Clostridium difficile toxin B.

I Just, J. Selzer, M Wilm … (1995)

Toxin A and B, the major virulence factors of Clostridium difficile, are the causative agents of antibiotic-associated pseudomembranous colitis. In cultured cell lines their potent cytotoxicity results from their ability to induce disaggregation of the microfilament cytoskeleton. Toxin B acts on the low-molecular-mass GTPase RhoA, which is involved in the regulation of the actin cytoskeleton. We report here that toxin B catalyses the incorporation of up to one mole of glucose per mole of RhoA at the amino acid threonine at position 37. The modification was identified and localized by tandem electrospray mass spectrometry. UDP-glucose selectively serves as cosubstrate for the monoglucosylation reaction catalysed by toxin B. Microinjection of RhoA previously glucosylated by toxin B into monolayer cells caused disaggregation of actin filaments, indicating a dominant-negative activity of glucosylated RhoA.

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The Clostridium sporulation programs: diversity and preservation of endospore differentiation.

Mohab Al-Hinai, Shawn W Jones, Eleftherios T Papoutsakis (2015)

Bacillus and Clostridium organisms initiate the sporulation process when unfavorable conditions are detected. The sporulation process is a carefully orchestrated cascade of events at both the transcriptional and posttranslational levels involving a multitude of sigma factors, transcription factors, proteases, and phosphatases. Like Bacillus genomes, sequenced Clostridium genomes contain genes for all major sporulation-specific transcription and sigma factors (spo0A, sigH, sigF, sigE, sigG, and sigK) that orchestrate the sporulation program. However, recent studies have shown that there are substantial differences in the sporulation programs between the two genera as well as among different Clostridium species. First, in the absence of a Bacillus-like phosphorelay system, activation of Spo0A in Clostridium organisms is carried out by a number of orphan histidine kinases. Second, downstream of Spo0A, the transcriptional and posttranslational regulation of the canonical set of four sporulation-specific sigma factors (σ(F), σ(E), σ(G), and σ(K)) display different patterns, not only compared to Bacillus but also among Clostridium organisms. Finally, recent studies demonstrated that σ(K), the last sigma factor to be activated according to the Bacillus subtilis model, is involved in the very early stages of sporulation in Clostridium acetobutylicum, C. perfringens, and C. botulinum as well as in the very late stages of spore maturation in C. acetobutylicum. Despite profound differences in initiation, propagation, and orchestration of expression of spore morphogenetic components, these findings demonstrate not only the robustness of the endospore sporulation program but also the plasticity of the program to generate different complex phenotypes, some apparently regulated at the epigenetic level.

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The genome and transcriptomes of the anti-tumor agent Clostridium novyi-NT.

Xiaosong Zhang, Shibin Zhou, L. Calero Diaz … (2006)

Bacteriolytic anti-cancer therapies employ attenuated bacterial strains that selectively proliferate within tumors. Clostridium novyi-NT spores represent one of the most promising of these agents, as they generate potent anti-tumor effects in experimental animals. We have determined the 2.55-Mb genomic sequence of C. novyi-NT, identifying a new type of transposition and 139 genes that do not have homologs in other bacteria. The genomic sequence was used to facilitate the detection of transcripts expressed at various stages of the life cycle of this bacterium in vitro as well as in infections of tumors in vivo. Through this analysis, we found that C. novyi-NT spores contained mRNA and that the spore transcripts were distinct from those in vegetative forms of the bacterium.

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Author and article information

Contributors

Chang-Gi Jeong: jcg0102@gmail.com

Byoung-Joo Seo: byoungjooseo@gamil.com

Salik Nazki: saliknazki@gmail.com

Byung Kwon Jung: bkjung@knu.ac.kr

Amina Khatun: amina.vet.sau.bd@gmail.com

Myeon-Sik Yang: 111@jbnu.ac.kr

Seung-Chai Kim: leesor2@jbnu.ac.kr

Sang-Hyun Noh: livensh@gmail.com

Jae-Ho Shin: jhshin@knu.ac.kr

Bumseok Kim: bskims@jbnu.ac.kr

Won-Il Kim:

ORCID: http://orcid.org/0000-0002-0465-0794

kwi0621@jbnu.ac.kr

Journal

Journal ID (nlm-ta): BMC Vet Res

Journal ID (iso-abbrev): BMC Vet. Res

Title: BMC Veterinary Research

Publisher: BioMed Central (London )

ISSN (Electronic): 1746-6148

Publication date (Electronic): 6 May 2020

Publication date PMC-release: 6 May 2020

Publication date Collection: 2020

Volume: 16

Electronic Location Identifier: 127

Affiliations

[1 ]GRID grid.411545.0, ISNI 0000 0004 0470 4320, College of Veterinary Medicine, Jeonbuk National University, ; 79 Gobong-ro, Iksan, Jeonbuk 54596 Republic of Korea

[2 ]GRID grid.258803.4, ISNI 0000 0001 0661 1556, School of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, ; Daegu, 41566 Republic of Korea

[3 ]GRID grid.462795.b, ISNI 0000 0004 0635 1987, Department of Pathology, Faculty of Animal Science and Veterinary Medicine, , Sher-e-Bangla Agricultural University, ; Sher-e-Bangla Nagar, Dhaka, 1207 Bangladesh

[4 ]GRID grid.497677.c, ISNI 0000000406477176, MSD Animal Health Korea Ltd., ; Seoul, 04637 Republic of Korea

Author information

Won-Il Kim http://orcid.org/0000-0002-0465-0794

Article

Publisher ID: 2349

DOI: 10.1186/s12917-020-02349-9

PMC ID: 7203850

PubMed ID: 32375805

SO-VID: 6ccbed2a-d8c7-4a15-a186-8816982affca

License:

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

History

Date received : 14 January 2020

Date accepted : 29 April 2020

Funding

Funded by: Rural Development Administration

Award ID: PJ010530

Award Recipient : Won-Il Kim

Custom metadata

ScienceOpen disciplines: Veterinary medicine

Keywords: anaerobe,clostridium novyi,isolation,characterization,whole-genome sequencing

Data availability:

ScienceOpen disciplines: Veterinary medicine

Keywords: anaerobe, clostridium novyi, isolation, characterization, whole-genome sequencing

Characterization of Clostridium novyi isolated from a sow in a sudden death case in Korea

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Horses and Humans Research Foundation

Most cited references 27

Glucosylation of Rho proteins by Clostridium difficile toxin B.

The Clostridium sporulation programs: diversity and preservation of endospore differentiation.

The genome and transcriptomes of the anti-tumor agent Clostridium novyi-NT.

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