Evaluation of representativity of the acetic acid bacteria species identified by culture-dependent method during a traditional wine vinegar production

• Published in: Food research international 2013-04, Vol.51 (1), p.404-411
• Main Authors: Vegas, Carlos, González, Ángel, Mateo, Estibaliz, Mas, Albert, Poblet, Montse, Torija, Maria Jesús
• Format: Article
• Language: English
• Subjects: Acetic acid; acetic acid bacteria; Acetobacter; Bacteria; Biological and medical sciences; correlation; culture media; DGGE-PCR; Fermented food industries; Food industries; Foods; Fundamental and applied biological sciences. Psychology; genes; Gluconacetobacter; Gluconacetobacter europaeus; Gluconacetobacter intermedius; Gluconacetobacter xylinus; Media; Microorganisms; phylogeny; Plating; quantitative polymerase chain reaction; ribosomal RNA; RT-PCR; TaqMan-MGB probes; Vinegar; Vitaceae; wine vinegars; Wines; Wines and vinegars
• ISSN: 0963-9969; 1873-7145
• DOI: 10.1016/j.foodres.2012.12.055

The low recovery of acetic acid bacteria (AAB) on solid media is one of the biggest hurdles to studying the microbiota present in vinegar production. For this reason, we carried out a comparative analysis between culture-dependent and independent methods to evaluate if the main AAB species isolated by plating are representative of the microbiota present. 265 isolates from two different culture media were identified and characterized by the 16S rRNA gene PCR-RFLP, ERIC-PCR and (GTG)5-rep-PCR. Additionally, a phylogenetic analysis using the 16S rRNA gene was performed. Finally, qualitative and quantitative culture-independent techniques (DGGE-PCR, 16S rRNA gene PCR-RFLP and Real Time PCR) were used for detecting and quantifying the AAB directly from vinegar. In general, the main species recovered by culture-dependent method were the same as those detected by culture-independent methods, confirming that plating could provide a valid initial approach for determining the diversity of the microbiota. The species detected by both methods were Gluconacetobacter europaeus, Gluconacetobacter xylinus and Gluconacetobacter intermedius. Furthermore, plating allowed us to detect some putative new Gluconacetobacter species that would have remained undetected with culture-independent techniques. ► The species recovered by both culture-dependent and independent methods are the same. ► Culture-dependent method did not allow a proper quantification of these species. ► No culture-independent method allows quantification and identification simultaneously.