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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(5): 715-724
DOI: https://doi.org/10.20546/ijcmas.2019.805.084


Characterization of Toxins in the Culture Supernatants of Clostridium difficile and their Immunogenic Potential in Mice
Parishmita Hazarika1, Rajeev K. Sharma1, Girindra K. Saikia1, Durlav P. Bora1*, Luit M. Borkalita2, Sophia M. Gogoi1, Nirab K. Deuri2 and Ritam Hazarika2
1Department of Microbiology
2Department of Animal Biotechnology, College of Veterinary Science, Assam Agricultural University, Khanapara Campus, Guwahati, Pin 781022, Assam, India
*Corresponding author
Abstract:

The Clostridium difficile toxins, toxin A and Toxin B were characterised in respect to the influence of glucose and incubation period on release of toxins, cytotoxic activities of the toxins in Verocell line and the immunogenic potential of C. difficile toxoids. A gradual increase in protein concentrations was observed in the cell free supernatants of toxin A and B positive C. difficile isolate with increase in incubation period and found to reach the highest at 48 hr of incubation, i.e., 5.24 µg/µl and 5.06 µg/µl, respectively. Release of C. difficile toxins in the culture supernatants was suppressed by glucose supplement in the nutrient media. Both the partially purified toxins were found to be cytotoxic for vero cells at both 1:10 and 1:100 dilutions. Cytotoxic activity of toxin B was more prominent than toxins A. However, similar immune-protective efficacy (100.0%) was exhibited by both the toxoid preparations (Toxoid A and B) in immunized animals against homologous challenge with 6.0x108 CFU, while 75.0 percent protection against 9.0x108 CFU of homologous strains of C. difficile was observed in both the groups, immunized separately with toxoid A and B. The affected mice, following challenge with 9.0x108 CFU dose could show clinical symptoms, suggestive of intestinal disorder, without mortality. However, all the mice of control group died on 48 hr of post challenge with both 6.0 x 108 and 9.0x108CFU / dose of homologous strain of C. difficile.


Keywords: Clostridium difficile tcdA, tcdB Toxins Immunogenic Potential in Mice
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How to cite this article:

Parishmita Hazarika, Rajeev K. Sharma, Girindra K. Saikia, Durlav P. Bora, Luit M. Borkalita, Sophia M. Gogoi, Nirab K. Deuri and Ritam Hazarika. 2019. Characterization of Toxins in the Culture Supernatants of Clostridium difficile and their Immunogenic Potential in Mice.Int.J.Curr.Microbiol.App.Sci. 8(5): 715-724. doi: https://doi.org/10.20546/ijcmas.2019.805.084