Investigation of Optimum Conditions of Sporogenesis and Cultivation of Clostridium novyi: cause of Gas gangrene

Document Type : Full Research Paper

Authors

1 Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran

2 Specialized Clostridia research laboratory, department of anaerobic vaccine research and production, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Alborz, Iran

3 Departments of Hepatitis and HIV, Pasteur Institute of Iran (IPI), Tehran, Iran

Abstract

Clostridium novyi is endospore- forming, obligate anaerobic bacteria of the class clostridia. Cultivation of this bacterium in common culture is one of the problems with this bacterium. It is pathogenic, causing a wide variety of diseases in man and animals the spore of this bacterium has also received much attention in the treatment of cancer due to its ability to grow in anaerobic conditions inside solid tumors. In this study, the optimum growth conditions for vegetative and spore form was investigated. Bacterial culture was investigated in different solid media including BHI, Blood Agar, F medium and NP medium in vegetative form. Also, a suitable spore media was introduced, the spore purification method, an easy method for bacterial count and spore aspiration were investigated. The best rich media for C.novyi in vegetative form are F and NP media. That is, by adding horse blood to these media culture, the growth be lux but is not a sufficient factor for growth. There was an almost linear relationship between bacterial absorption and bacterial count for Clostridium novyi. Also, the difference in the number of spores germination which heating at 80 ° C for 15 minutes was significant.   

Keywords

References

1- Aktories, K. and T. Jank. Section. 2015. 15 - Large clostridial cytotoxins modifying small GTpases: structural aspects. 426-440. In: Alouf J., Ladant D., Popoff MR, editors. The Comprehensive Sourcebook of Bacterial Protein Toxins (Fourth Edition). Academic Press. Boston.
2- Aquino, P. L., F. S. Fonseca, O. D. Mozzer, R. C. Giordano and R. Sousa, Jr. 2016. Optimization of the Production of Inactivated Clostridium novyi Type B Vaccine Using Computational Intelligence Techniques. Appl Biochem Biotechnol 179: 895-909.
3- Ataei Daryaei, F., R. Pilehchian Langroudi and B. Eimani. 2017. Production and Evaluation of a New Recombinant Cloning Vector for the Clostridium novyi Type B Vaccine Strain Alpha Toxin Gene.
4- B Moore, W. 1968. Solidified Media Suitable for the Cultivation of Clostridium novyi Type B.
5- Dang, L. H., C. Bettegowda, D. L. Huso, K. W. Kinzler and B. Vogelstein. 2001. Combination bacteriolytic therapy for the treatment of experimental tumors. Proceedings of the National Academy of Sciences 98: 15155.
6- Dang, L. H., C. Bettegowda, D. L. Huso, K. W. Kinzler and B. Vogelstein. 2001. Combination bacteriolytic therapy for the treatment of experimental tumors. Proc Natl Acad Sci U S A 98: 15155-15160.
7- Eklund, M. W., F. T. Poysky, M. E. Peterson and J. A. Meyers. 1976. Relationship of bacteriophages to alpha toxin production in Clostridium novyi types A and B. Infection and immunity 14: 793-803.
8- M Aronoff, D. 2013. Clostridium novyi, sordellii, and tetani: Mechanisms of disease.
9- Nyaoke, A. C., M. A. Navarro, J. Beingesser and F. A. Uzal. 2018. Infectious necrotic hepatitis caused by Clostridium novyi type B in a horse: case report and review of the literature. J Vet Diagn Invest 30: 294-299.
10- Underwood, W. J., R. Blauwiekel, M. L. Delano, R. Gillesby, S. A. Mischler and A. Schoell. Section. 2015. Chapter 15 - Biology and Diseases of Ruminants (Sheep, Goats, and Cattle). 623-694. In: Fox JG., Anderson LC., Otto GM., Pritchett-Corning KR., Whary MT, editors. Laboratory Animal Medicine (Third Edition). Academic Press. Boston.
11- Wang, L., Q. Wang, X. Tian and X. Shi. 2018. Learning from Clostridium novyi-NT: How to defeat cancer. Journal of Cancer Research and Therapeutics 14: 1-6.
12- Watanabe, N., K. Kobayashi, G. Hashikita, Y. Taji, N. Ishibashi, S. Sakuramoto, K. Mitsutake, K. Ikebuchi and Y. Ebihara. 2019. Hepatic gas gangrene caused by Clostridium novyi. Anaerobe 57: 90-92.
13- Yang, W. W., E. N. Crow-Willard and A. Ponce. 2009. Production and characterization of pure Clostridium spore suspensions. Journal of Applied Microbiology 106: 27-33.
14-پیله چیان لنگرودی, ر. and ا. ر. جباری. 2013. تولید واکسن براکسی با استفاده از محیط کشت غنی شده و فرمانتور. مجله تحقیقات دامپزشکی (Journal of Veterinary Research) 68: 297-304.

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History

  • Receive Date: 06 August 2019
  • Accept Date: 26 November 2019

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