Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 503314
Title Use of propidium monoazide for selective profiling of viable microbial cells during Gouda cheese ripening
Author(s) Erkus, Oylum; Jager, Victor C.L. de; Geene, Renske T.C.M.; Alen-Boerrigter, Ingrid van; Hazelwood, Lucie; Hijum, Sacha A.F.T. van; Kleerebezem, Michiel; Smid, Eddy J.
Source International Journal of Food Microbiology 228 (2016). - ISSN 0168-1605 - p. 1 - 9.
DOI https://doi.org/10.1016/j.ijfoodmicro.2016.03.027
Department(s) Microbiological Laboratory
FB OS Universitair Sportcentrum
Host Microbe Interactomics
VLAG
WIAS
Food Microbiology Laboratory
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) Cheese - Dairy - Food fermentation - Metagenomics - Microbial community profiling - Propidium monoazide
Abstract

DNA based microbial community profiling of food samples is confounded by the presence of DNA derived from membrane compromised (dead or injured) cells. Selective amplification of DNA from viable (intact) fraction of the community by propidium monoazide (PMA) treatment could circumvent this problem. Gouda cheese manufacturing is a proper model to evaluate the use of PMA for selective detection of intact cells since large fraction of membrane compromised cells emerges as a background in the cheese matrix during ripening. In this study, the effect of PMA on cheese community profiles was evaluated throughout manufacturing and ripening using quantitative PCR (qPCR). PMA effectively inhibited the amplification of DNA derived from membrane compromised cells and enhanced the analysis of the intact fraction residing in the cheese samples. Furthermore, a two-step protocol, which involves whole genome amplification (WGA) to enrich the DNA not modified with PMA and subsequent sequencing, was developed for the selective metagenome sequencing of viable fraction in the Gouda cheese microbial community. The metagenome profile of PMA treated cheese sample reflected the viable community profile at that time point in the cheese manufacturing.

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