Development of a species‐specific PCR assay for identification of the strictly anaerobic bacterium Selenomonas lacticifex found in biofilm‐covered surfaces in brewery bottling halls

• Published in: Journal of applied microbiology 2014-11, Vol.117 (5), p.1328-1335
• Main Authors: Felsberg, J, Jelínková, M, Kubizniaková, P, Matoulková, D
• Format: Article
• Language: English
• Subjects: Bacteria; Beer - microbiology; beer spoilage; beers; Bioassays; biofilm; Biofilms; Biological and medical sciences; bottling; Breweries; brewers yeast; brewing; dissolved oxygen; DNA Primers; Fundamental and applied biological sciences. Psychology; Megasphaera; Microbiology; packaging; PCR; Pectinatus; Polymerase chain reaction; Polymerase Chain Reaction - methods; ribosomal DNA; risk; RNA, Ribosomal, 16S - genetics; Saccharomyces cerevisiae; secondary contamination; Selenomonas - isolation & purification; Selenomonas - physiology; Selenomonas lacticifex; Selenomonas ruminantium; Species Specificity; strictly anaerobic bacteria; Zymophilus
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.12610

AIMS: In recent years, beer‐spoilage cases from strictly anaerobic bacteria have risen in frequency, in connection with the production of non‐pasteurized, non‐alcohol and low‐alcoholic beers and with the lowering of dissolved oxygen in the packaged beer. Selenomonas lacticifex, found in brewer's yeast and in biofilms covering some surfaces in brewery bottling area, is considered to be a beer‐spoilage organism. This study aims to develop S. lacticifex‐specific PCR assay. The objective of this study was also evaluation of the specificity and reproducibility of the developed PCR assay in real brewery samples. METHODS AND RESULTS: Three primers (one forward and two reverse) were designed for identification of the strictly anaerobic bacterium S. lacticifex on the basis of the species‐specific sequences of the 16S rDNA region. The specificity of the primers was tested against 44 brewery‐related non‐target micro‐organisms that could potentially occur in the same brewery specimens. None of the primer pairs amplified DNA from any of the non‐S. lacticifex strains tested including genera from the same family (Pectinatus, Megasphaera, Zymophilus) and the closely related species Selenomonas ruminantium, showing thus 100% specificity. CONCLUSIONS: The PCR assay developed in this study enables the detection of the strictly anaerobic bacterium S. lacticifex in real brewery samples including pitching yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: Selenomonas lacticifex‐specific PCR assay developed in this study allows for the extension of the spectra of detected beer‐spoilage micro‐organisms in brewing laboratories and thus lowering the risk of contamination of the final product.