RESUMO
Lactic acid bacteria collected from artisanal farmhouses were characterized using a polyphasic approach. Phenotypic methods including biochemical assays, ribosomal DNA restriction analysis and 16S rDNA sequence analysis were performed. This approach provides accuracy for identification, and helps to avoid the loss of natural biodiversity including potentially valuable strains.
Assuntos
Laticínios/microbiologia , Ácido Láctico/metabolismo , Lactobacillales/classificação , Lactobacillales/isolamento & purificação , Animais , Técnicas de Tipagem Bacteriana , Bovinos , Análise por Conglomerados , DNA Ribossômico/química , DNA Ribossômico/genética , Cabras , Lactobacillales/genética , Lactobacillales/fisiologia , Dados de Sequência Molecular , Filogenia , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , OvinosRESUMO
The occurrence of spore-forming bacteria in powdered milk is of concern to the dairy industry due to potential deleterious effects including those resulting from proteolytic and lipolytic activities. Twenty-two powdered milk samples representative of spring and summer production obtained from Uruguayan retail stores were analyzed for type and number of thermophilic and spore-forming bacterial species. Bacillus licheniformis isolates were found to be the most prominent milk powder contaminant followed by Anoxybacillus flavithermus representing 71.5 to 84% of the total microflora. Geobacillus stearothermophilus, however, was not found. B. licheniformis strains F and G were both found in this study but strain F was the prevalent isolate representing 98.9% of the total isolates of this species. A. flavithermus isolates corresponded to strain C in accordance with 16S rRNA gene sequence analysis, however, in contrast with other reports, the RAPD profiles showed three characteristic bands at approximately 650, 1000 and 1650 bp, but lacking a band at 1250 bp. A third group of isolates was identified corresponding to members of a Bacillus subtilis group and Bacillus megaterium. Isolates designated B. licheniformis, A. flavithermus, B. megaterium and the B. subtilis group represented 89.1 to 93.6% of those analyzed, and depended on previous heat treatment and incubation temperatures of the plates. The remaining isolates were Bacillus pumilus and unidentified spore-formers.