16S rRNA in situ Hybridization Followed by Flow Cytometry for Rapid Identification of Acetic Acid Bacteria Involved in Submerged Industrial Vinegar Production

Janja Trček1,2*, Luka Lipoglavšek3 and Gorazd Avguštin3


1Department of Biology, Faculty of Natural Sciences and Mathematics, University of Maribor, Koroška cesta 160, SI-2000 Maribor, Slovenia
2Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova ulica 17, SI-2000 Maribor, Slovenia
3Department of Animal Science, Biotechnical Faculty, University of Ljubljana, Groblje 3, SI-1230 Domžale, Slovenia

Article history:
Received  December 20, 2014
Accepted October 13, 2015

Key words:
microbiota, vinegar, acetic acid bacteria, Acetobacter, Gluconobacter, Gluconacetobacter, Komagataeibacter, 16S rRNA probe, in situ hybridization, flow cytometry


Acetic acid bacteria are involved in many biotechnological processes such as vitamin C, gluconic acid, miglitol or acetic acid production, and others. For a technologist trying to control the industrial process, the ability to follow the microbiological development of the process is thus of importance. During the past few years hybridization in a combination with flow cytometry has oft en been used for this purpose. Since vinegar is a liquid, it is an ideal matrix for flow cytometry analysis. In this work we have constructed a specific probe for highly acetic acid-resistant species of the acetic acid bacteria and a protocol for in situ hybridization, which in combination with flow cytometry enables direct monitoring of bacteria producing vinegar with >10 % of acetic acid. The approach was successfully applied for monitoring microbiota during industrial vinegar production.




*Corresponding author:  email3
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