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Sci Rep. 2018 Jul 30;8(1):11446. doi: 10.1038/s41598-018-28949-y.

Discovery and control of culturable and viable but non-culturable cells of a distinctive Lactobacillus harbinensis strain from spoiled beer.

Liu J1,2, Deng Y3, Li L4,5,6, Li B1,7, Li Y8, Zhou S9, Shirtliff ME10, Xu Z11,12,13, Peters BM2.

Author information

1
School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, China.
2
Department of Clinical Pharmacy, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN, 38163, USA.
3
College of Food Science and Engineering, Qingdao Agricultural University, Qingdao, 266109, P.R. China.
4
School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, China. felinli@scut.edu.cn.
5
School of Chemical Engineering and Energy Technology, Dongguan University of Technology, Dongguan, 523808, China. felinli@scut.edu.cn.
6
Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Guangzhou, 510640, China. felinli@scut.edu.cn.
7
Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Guangzhou, 510640, China.
8
Department of Cell Biology, Harvard Medical School, Boston, MA, 02115, USA.
9
School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, China.
10
Department of Microbial Pathogenesis, School of Dentistry, University of Maryland, Baltimore MD, MA, 21201, USA.
11
School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, China. zhenbo.xu@hotmail.com.
12
College of Food Science and Engineering, Qingdao Agricultural University, Qingdao, 266109, P.R. China. zhenbo.xu@hotmail.com.
13
Department of Microbial Pathogenesis, School of Dentistry, University of Maryland, Baltimore MD, MA, 21201, USA. zhenbo.xu@hotmail.com.

Abstract

Occasional beer spoilage incidents caused by false-negative isolation of lactic acid bacteria (LAB) in the viable but non-culturable (VBNC) state, result in significant profit loss and pose a major concern in the brewing industry. In this study, both culturable and VBNC cells of an individual Lactobacillus harbinensis strain BM-LH14723 were identified in one spoiled beer sample by genome sequencing, with the induction and resuscitation of VBNC state for this strain further investigated. Formation of the VBNC state was triggered by low-temperature storage in beer (175 ± 1.4 days) and beer subculturing (25 ± 0.8 subcultures), respectively, and identified by both traditional staining method and PMA-PCR. Resuscitated cells from the VBNC state were obtained by addition of catalase rather than temperature upshift, changing medium concentration, and adding other chemicals, and both VBNC and resuscitated cells retained similar beer-spoilage capability as exponentially growing cells. In addition to the first identification of both culturable and VBNC cells of an individual L. harbinensis strain from spoiled beer, this study also for the first time reported the VBNC induction and resuscitation, as well as verification of beer-spoilage capability of VBNC and resuscitated cells for the L. harbinensis strain. Genes in association with VBNC state were also identified by the first genome sequencing of beer spoilage L. harbinensis. The results derived from this study suggested the contamination and spoilage of beer products by VBNC and resuscitated L. harbinensis strain BM-LH14723.

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