title: Acetic acid bacteria from biofilm of strawberry vinegar visualized by microscopy and detected by complementing culture-dependent and culture-independent techniques.
abstract: Acetic acid bacteria (AAB) usually develop biofilm on the air-liquid interface of the vinegar elaborated by traditional method. This is the first study in which the AAB microbiota present in a biofilm of vinegar obtained by traditional method was detected by pyrosequencing. Direct genomic DNA extraction from biofilm was set up to obtain suitable quality of DNA to apply in culture-independent molecular techniques. The set of primers and TaqMan--MGB probe designed in this study to enumerate the total AAB population by Real Time--PCR detected between 8 × 10(5) and 1.2 × 10(6) cells/g in the biofilm. Pyrosequencing approach reached up to 10 AAB genera identification. The combination of culture-dependent and culture-independent molecular techniques provided a broader view of AAB microbiota from the strawberry biofilm, which was dominated by Ameyamaea, Gluconacetobacter, and Komagataeibacter genera. Culture-dependent techniques allowed isolating only one genotype, which was assigned into the Ameyamaea genus and which required more analysis for a correct species identification. Furthermore, biofilm visualization by laser confocal microscope and scanning electronic microscope showed different dispositions and cell morphologies in the strawberry vinegar biofilm compared with a grape vinegar biofilm.
authors.affiliation: Biotecnologia Enológica, Dept. Bioquímica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, C/Marcellí Domingo s/n, 43007 Tarragona, Spain.
authors.forename: Maria José
authors.initials: MJ
authors.lastname: Valera
authors.affiliation: Biotecnologia Enológica, Dept. Bioquímica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, C/Marcellí Domingo s/n, 43007 Tarragona, Spain.
authors.forename: Maria Jesús
authors.initials: MJ
authors.lastname: Torija
authors.affiliation: Biotecnologia Enológica, Dept. Bioquímica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, C/Marcellí Domingo s/n, 43007 Tarragona, Spain.
authors.forename: Albert
authors.initials: A
authors.lastname: Mas
authors.affiliation: Departamento de Inmunología, Microbiología y Parasitología, Facultad de Medicina y Odontología, Universidad del País Vasco/Euskal Herriko Unibertsitatea, UPV/EHU, Barrio Sarriena s/n, 48940 Leioa, Spain. Electronic address: estibaliz.mateo@ehu.es.
authors.forename: Estibaliz
authors.initials: E
authors.lastname: Mateo
mesh_terms: D019342:Acetic Acid / Q000032:analysis / Q000378:metabolism*; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D018441:Biofilms; D005285:Fermentation; D031985:Fragaria / Q000378:metabolism / Q000382:microbiology*; D008969:Molecular Sequence Data; D010802:Phylogeny
chemical_list: D019342:Acetic Acid
keywords: 16S rRNA gene; Next-generation sequencing; PCR-DGGE; Vinegar; Wine