Acetobacter aceti fast identification by Real Time PCR in spoiled wine samples

Attila Kántor

Abstract


Wine is a beverage that made from grape berries. However, without beneficial bacteria, we would not produce good wine. But very often wines contain acetic acid bacteria, which are undesirable in winemaking process. Acetic acid bacteria as known as a vinegar bacteria are Gram-negative, aerobic, rod-shaped and ubiquitous bacteria. This study was focused on species of acetic acid bacteria, specifically Acetobacter aceti that make spoilage in wine.The aim of our study was the identification of Acetobacter aceti in spoiled red wine samples, with plate dilution method on agar plates and using sensitive Real-time PCR (qPCR) method. We cultivated Acetobacter aceti on GYC agar at 30°C, 48h. The one of main objective in the present work was the test fast, sensitive and reliable technique such as quantitative Real-time PCR and detecting the presence of Acetobacter aceti in wine samples with positive Acetobacteraceti control on amplification plot and melting curve. The next objective before  qPCR analysis was DNA extraction from wine samples incubated for one week at 28°C aerobically. We used five different red wine samples for this experiment: Alibernet 2013, Blaufränkisch 2013, Cabernet Sauvignon 2013, Dunaj 2012 and Saint-Laurent 2012. Next we extracted DNA from wine samples and from pure Acetobacter aceti CCM 3620T strain purchased from Czech collection of microorganisms in Brno. Susceptibility ofAcetobacter aceti was varied in different isolates from 102 to 107 CFU.mL-1. The number of Acetobacter cells on GYC medium ranged from 4.05 to 4.83log CFU.mL-1 in differentwine samples.The higher number of Acetobacter cells (4.83 log CFU.mL-1) was found in Cabernet Sauvignon 2013 wine.

Keywords


Acetobacter aceti; wine; Real Time PCR; spoilage

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Scientific Papers Animal Science and Biotechnologies publish open access articles under the terms of the Creative Commons Attribution 4.0 Licence (read full legal code).


LUCRARI STIINTIFICE ZOOTEHNIE SI BIOTEHNOLOGII (SCIENTIFIC PAPERS ANIMAL SCIENCE AND BIOTECHNOLOGIES)

ISSN print 1841 - 9364
ISSN online 2344 - 4576
ISSN-L  1841 - 9364
(former ISSN 1221-5287, E-ISSN 1841-9364)

PUBLISHER: AGROPRINT Timisoara, Romania
PAPER ACCESS: Full text articles available for free
FREQUENCY: Semiannual
PUBLICATION LANGUAGE: English
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Banat´s University of Agricultural Sciences and Veterinary Medicine "King Michael I of Romania" from Timisoara
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