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Identification of bacterial contaminants of the cachaça ferment by sequencing of the 16S rDNA Cachaça is a typical Brazilian spirit made from sugar cane fermented mainly by Saccharomyces cerevisiae. The production is mostly artisanal, and there is no microbiological control during the fermentation process. The objective of this study was to assess the bacterial community associated with the ferment used in the production of cachaça. Four ferment samples were collected from an artisanal still. The first one (NA), was collected one year previously to the other three and constituted a control reference. The remaining three samples were collected at the end of the first day of fermentation (NP), and fifteen (NS) and thirty days (NT) after using this same ferment. A total of 587 16S rDNA sequences were analyzed, being 81 sequences from the NA sample, 177 from NP, 159 from NS, and 170 from the NT sample. Sequence analyses revealed the presence of 17 genus and 27 species plus 27 unknown species. One hundred and seventy operational taxonomic units (OTUs) were identified using the DOTUR software using a cut-off evolutionary distance of 0.03. Forty three were identified in the control (NA) sample, 38 in the NP, 57 in NS, and 38 in the third (NT) sample. Of the 170 OTUs, only seventeen were detected twice in different samples and only one, identified as Lactobacillus hilgardii, was identified thrice, indicating a dynamic nature of the bacterial community during the fermentation process. Statistical analysis using the software S-LIBSHUFF indicated significant differences in the bacterial composition among samples. Our analyses also allowed to identify several bacteria not yet described as contaminants of the cachaça ferment, such as Weissella cibaria, Leuconostoc citreum and some Lactobacillus species. In addition, several unknown bacteria were also detected. The results revealed a much larger bacterial contaminant community present in the fermentative process of cachaça than previously reported with heterofermentative ability to produce secondary compounds which probably influence the quality of the final beverage. This is the first report on the utilization of the 16S rDNA sequencing method to assess the bacterial diversity of sugar cane fermentation for the production of cachaça.

Identificação de bactérias contaminantes de fermento de cachaça por seqüenciamento do gene 16S rDNA