Validation of the official control method based on Polymerase Chain Reaction (PCR) for identification of authorised probiotic yeast in animal feed

Systematic and Applied Microbiology
Renata G K LeuschnerGérard Bertin

Abstract

Council Directive 70/524/EEC regulates the application of probiotic (microorganisms) additives in feeding stuffs. In the present study a method for the differentiation and strain identification of authorised probiotic Saccharomyces cereviseae strains in feeding stuffs by Polymerase Chain Reaction (PCR) was validated. Four different samples of animal feeding stuffs containing yeast at levels between 10(5) to 10(7) CFU/g were examined. Samples were enumerated on chloramphenicol glucose yeast extract agar and colonies were selected from these plates for DNA extraction and subsequent analysis. The PCR method using delta sequence primers produced an 'amplified sequence polymorphism' characteristic for the test strain. Feeds supplemented with one of four probiotic yeast strains each were analysed by seven of nine invited laboratories. All laboratories returned valid results with the exception of one laboratory that had insufficiently separated bands on the gel. The method had a good reproducibility for probiotic yeast isolates from feed of all four authorised probiotic yeast strains (APYS) CBS 493.94, APYS CNCM 1-1079, APYS CNCM 1-1077, APYS NCYC SC47 and of a commercially available yeast reference strain, NCYC 81. The PCR method is ...Continue Reading

Citations

Mar 27, 2010·Journal of Applied Microbiology·N R BüchlS Scherer
Jul 12, 2005·Yeast
Oct 21, 2006·The Cochrane Database of Systematic Reviews·B RichterS H Ebrahim
Dec 17, 2008·Nutrition Research Reviews·G BoschM W A Verstegen
Feb 16, 2011·Journal of Animal Physiology and Animal Nutrition·S E ToddW H Hendriks

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