Pyrosequencing vs. culture-dependent identification to analyze lactic acid bacteria population associated to Chicha, a traditional maize-based fermented beverage from Northwestern Argentina

AZNAR ROSA; ELIZAQUÍVEL PATRICIA ; PÉREZ ALBA; YÉPEZ ALBA ; JIMÉNEZ EUGENIA; ARISTIMUÑO FICOSECO MARIA CECILIA; COCCONCELLI ; GRACIELA VIGNOLO

Valencia

Congreso; ECCO XXXIII Molecular Taxonomy: from biodiversity to biotechnology; 2014

European Culture Collections? Organisation

Traditional fermented foods and beverages areproduced by natural fermentation involving mixed populations of indigenousmicroorganisms. Among them lactic acid bacteria (LAB) are relevant players inthe process. Chicha is a traditional maize-based fermented alcoholic beverageproduced following artisanal and ancestral procedures in the Andean regions. Inthis study the diversity of LAB associated with chicha fermentation was analysedusing culture-dependent and culture-independent procedures to assessgenus/species composition. To that aim, samples from two chicha productionswere obtained from local producers at Maimará (chicha M) and Tumbaya (chicha T)in northwestern Argentina, each exhibiting particular manufacturing procedures.Samples corresponding to 10 production steps were analyzed by pyrosequencing ofthe V3-V5 region in the 16S rRNA gene to obtain a snapshot of the globalbacterial population. Sequence analysis and taxonomic assignment was performedusing the Ribosomal Enterococcus, Lactococcus, Streptococcus, Weissella,Leuconostoc and Lactobacillus while Pediococcuswas only detected in chichi M At species level, the highest diversity wasobserved for Lactobacillus fromchicha M and T, and Enterococcus fromchicha T. Predominant LAB species in chicha M were Lb. plantarum, Lb. rossiae, Lc. lactisand W. viridescens whereas inchicha T were E. ratti and Lc. mesenteroides Results showed most ofthe isolates from chicha M as belonging to Leuconostocgenus. (Lc. mesenteroidesand Lc. lactis)although Lactobacillus, Weissellaand Enterococcus were alsopresent. In contrast, chicha T exhibited the presence of Enterococcus and Leuconostoc,E. faecium being the mostrepresentative species during fermentation. Members of genera Lactococcus, Pediococcusand Streptococcuswere only detected by pyrosequencing in chicha M as with Weissella, Lactococcus, Lactobacillus and Streptococcus in chicha T. Pyrosequencing revealed the highestspecies diversity for Lactobacillus andEnterococcus in chicha M and Tsamples, respectively. Of them, Lactobacillusspecies were found by culturing in chicha M, but none in chicha T. Inconclusion, culture-dependent approach showed marked differences in LABdiversity from both chicha productions, whereas pyrosequencing, althoughprovided a more comductions, thus pointing to the species which may drive thefermentation process.