DETECTION OF TOXIGENIC STRAINS OF BACILLUS CEREUS IN MILK AND DAIRY PRODUCTS THROUGH POLYMERASE CHAIN REACTION

ABSTRACT

Fifty nine samples of raw milk, 60 samples of pasteurized milk, 24 cream, 39 butter, 27 curd, 19 peda, 21 paneer and 20 skim milk powder samples were analyzed for the presence of different Bacillus species and a total of 182 isolates were obtained. The isolates were identified as Bacillus subtilis (51), Bacillus megaterium (39), Bacillus cereus (28), Bacillus licheniformis (32), Bacillus coagulans (7), Bacillus sphaericus (8), Bacillus pumilus (9) and Bacillus circulans (8). All the 28 Bacillus cereus isolates obtained in the study were subjected to polymerase chain reaction amplification for the presence of 365 nucleotides and all the isolates formed a distinct PCR product of 365 bp, which confirmed that these isolates were Bacillus cereus. Only 2 out of 28 Bacillus cereus isolates were found to be enterotoxigenic. One isolate harboured all the five genes of B, B’, L1, L2 and ET of the HBL-ET complex whereas the other isolate contained B, L1, L2 and ET genes and the B’ component was missing. Both the toxigenic strains of Bacillus cereus produced similar discontinuous haemolysis pattern on blood agar and cytotoxic effect on Vero cell line.

KEYWORDS

REFERENCES

Beecher, D.J. and Wong, Amy C.L. (1994) Identification of haemolysin BL producing Bacillus cereus isolates by a discontinuous haemolytic pattern in blood agar. Appl. Environ. Microbiol., 60(5):1646-1651.
Buchanan, R.L., and Schultz, F.J. (1994) Comparison of the Tecra VIA kit, Oxoid BCET-RPLA kit and CHO cell culture assay for the detection of Bacillus cereus diarrhoeal enterotoxin. Lett. Appl. Microbiol., 19:353-356.
Buchanan, R.E., and Gibbons, N.E. (1994) Bergey’s manual of Determinative Bacteriology, 5th EDn. The willams and Wilkins Company, Baltimore, USA.
Christiansson, A., Naidu, A.S., Nilsson, I. Wadstrom, T. and Pettersso, H.E., (1989) Toxin production by Bcillus cereus dairy isolates in milk at low temperatures. Appl. Environ. Microbiol., 55:2595-2600.Fermanian, C., and Wong Amy C.L. (2000) Improved in vitro detection of haemolysin BL from Bacillus cereus. Int. J. Food Microbiaol., 57:1-8.
Giffel, M.C..te., Beumer, R.R. Bonestroo. M.H. and Rombouts. F.M., (1996) Incidence and characterization of Bacillus cereus in two dairy processing plants. Neth. Milk and Dairy J., 50(4): 479-492.
Maniatis, T. Fritsch, E.F. and Sambrook, I. (1982) Molecular Cloning. A Laboratory Manual. Cold Springs Harbor Laboratory, New York, US.
Mantynen, V., and Lindstrom, K. (1998) A rapid PCR-based DNA test for enterotoxic Bacillus cereus. Appl. Environ. Microbiol., 64:1634-1639.
Radhika, B., Padmapriya, B.P., Chandrasekar, A., Keshava, N. and Varadaraj, M.C. (2002) Detection of Bacillus cereus in foods by colony hybridization using PCR-generated probe and characterization of isolates for toxins by PCR. Int. J. Food Microbiol., 74:131-138.
Snedecor. G.W. and Cochran, E.G., (1989) Statistical Methods. 8th Ed. The lowa State University Press, Ames, lowa, USA.
Svensson, B., Monthan, A., Guinebretiere, M.H. Nguyen-the and Christiansson, A. (2007) Toxin production potential and the detection of toxin genes strains of the Bacillus cereus group isolated along the dairy production chain. Int. Dairy J, 17:1201-1208.
Torkar, K.G., and Mozina S.S. (2001). Determination of Bacillus cereus and its toxins from milk and milk products. Periodicum Biologorum, 103(2):169-173.
Veld. in’t P.H., Ritmeester, W.S., Delfgou-van Asch E.H.M., Dufrenne, J.B., Wernars, K., Smit, E. and Van Leusden, F.M. (2001). Detection of genes encoding for enterotoxins and determination of the production of enterotoxins by HBL blood plates and immunoassays of psychrotrophic strains of Bacillus cereus isolated from pasteurized milk. Int. J. Food Microbiol., 64(1/2)63-70.
Yamada. S., Ohashi, E., Agata, N. and Venkateswaran, K. (1999). Cloning and nucleotide sequence analysis of gyrB of Bacillus cereus, B. thuringiensis, B. mycoides and B.anthracis and their application to the detection of B.cereus in rice. Appl. Environ. Microbiol., 65:1483-1490.

Disclaimer :

The Asian Journal of Dairy and Food Research (AJDFR) follows a disclaimer policy that outlines the responsibilities of the journal, authors, reviewers, and readers. The policy is designed to ensure that all published content is accurate and meets ethical standards, and to protect the journal from any legal liability.

The AJDFR does not guarantee the accuracy, completeness, or reliability of the content published in the journal. The opinions expressed in published articles are those of the authors and do not necessarily reflect the views of the journal or its editorial board. The journal does not accept any responsibility for any errors or omissions in the content published in the journal.

Authors are responsible for ensuring that their work is original, accurate, and meets ethical standards. They are also responsible for obtaining permission to use copyrighted material and for providing appropriate attribution.

Reviewers are responsible for ensuring that the content of articles is accurate, original, and meets ethical standards. They are also responsible for maintaining confidentiality and disclosing any conflicts of interest.

Readers are responsible for evaluating the content of articles and for making their own judgments about the accuracy and reliability of the information presented. They are also responsible for complying with copyright laws and ethical standards when using the content of articles.

AJDFR is committed to ensuring that all published content meets ethical standards and that all parties involved in the publishing process understand their responsibilities. The disclaimer policy is designed to protect the journal from any legal liability and to ensure that readers can rely on the accuracy and reliability of the content published in the journal.

Copyright :

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Asian Journal of Dairy and Food Research

DETECTION OF TOXIGENIC STRAINS OF BACILLUS CEREUS IN MILK AND DAIRY PRODUCTS THROUGH POLYMERASE CHAIN REACTION

ABSTRACT

KEYWORDS

REFERENCES

Reviewed By

In this Article

APC

Publish With US

Become a Reviewer/Member

Open Access

Products and Services

Support and Policies

Editorial Board