Characterization of dominant lactic acid bacteria isolated from São Jorge cheese, using biochemical and ribotyping methods.

AIMS: To identify, using phenotypic and genotypic methods, the dominant lactic acid bacteria (LAB) present in São Jorge cheese - one of the 11 Portuguese cheeses currently bearing an Appéllation d'Origine Protegée status. METHODS AND RESULTS: A total of 225 isolates from milk, curd and cheeses throughout ripening were identified to the genus level, 108 to the species level and ten to the strain level. Phenotypic methods indicated that lactobacilli, followed by enterococci, were the dominant bacteria. The most frequently isolated species were Lactobacillus paracasei, Lactobacillus rhamnosus, Enterococcus faecalis and Enterococcus faecium. Ribotyping differentiated three L. paracasei, two E. faecalis and one Lactobacillus plantarum types. Enterococcus spp. exhibited the highest esterase and beta-galactosidase activities among all isolates. CONCLUSIONS: The dominant LAB in São Jorge cheese are L. paracasei, L. rhamnosus, E. faecalis and E. faecium. Enterococcus likely plays a leading role upon acidification and aroma development in said cheese. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results support that a combination of conventional biochemical methods with genotypic methods allows for a thorough characterization and identification of isolates. Despite the limited number of isolates subject to molecular subtyping, a few specific Enterococcus and Lactobacillus strains were found that are promising ones for development of a starter culture. Hence, L. paracasei and E. faecalis are good candidates for a tentative starter culture, designed for manufacturing of São Jorge cheese at large - which takes advantage of actual isolates, in attempts to eventually standardize the quality of said cheese variety.

Detection and characterization of Listeria monocytogenes in Sao Jorge (Portugal) cheese production.

Listeria monocytogenes is a foodborne pathogen that can cause serious invasive disease in humans. Because human listeriosis cases have previously been linked to consumption of contaminated cheese, control of this pathogen throughout the cheese production chain is of particular concern. To understand the potential for L. monocytogenes transmission via São Jorge cheese, a Portuguese artisanal cheese variety that bears a Protected Denomination of Origin classification, 357 raw milk, curd, natural whey starter, and cheese samples representative of the production chain of this cheese were collected over one year and tested for the presence of L. monocytogenes and selected physicochemical parameters. Although neither L. monocytogenes nor other Listeria spp. were detected in whey, curd, or cheese samples, 2 of the 105 raw milk samples analyzed were positive for L. monocytogenes. These 2 raw milk isolates represented a ribotype that has previously been linked to multiple human listeriosis outbreaks and cases elsewhere, indicating the potential of these isolates to cause human listeriosis. On average, physicochemical parameters of São Jorge cheese ripened for 4 mo presented values that likely minimize the risk of L. monocytogenes outgrowth during ripening and storage (mean pH = 5.48; mean moisture = 37.79%; mean NaCl concentration = 4.73%). However, some cheese samples evaluated in this study were characterized by physicochemical parameters that may allow growth and survival of L. monocytogenes. Even though our results indicate that raw milk used for São Jorge cheese manufacture as well as finished products is rarely contaminated with L. monocytogenes, continued efforts to control the presence of this pathogen in the São Jorge cheese production chain are urged and are critical to ensure the safety of this product.

Manufacturing of fermented goat milk with a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus in a controlled bioreactor.

AIMS: This work was undertaken to study the feasibility and the characteristics of a fermented product made of goat milk, using a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus under controlled conditions, and to determine their survival in the fermented milk during refrigerated storage. METHODS AND RESULTS: Goat milk was inoculated with Lact. acidophilus and Bif. animalis mixed starter, fermented in a glass bioreactor with controlled temperature (37 degrees C) and anaerobiosis, and monitored for growth and acidification. The fermented milk was then stored for 10 days under refrigeration, and monitored daily for starter microflora survival and pH changes. Lact. acidophilus viable counts reached a maximum of 7.1 x 10(8) colony-forming units (CFU) ml(-1), and Bif. animalis a maximum of 6.3 x 10(7) CFU ml(-1) by 20 h of fermentation. During refrigerated storage, both strains exhibited a good survival, with viable numbers remaining essentially constant throughout the experiment, whereas the pH of the fermented milk dropped slightly. CONCLUSIONS: Mixed cultures of Bif. animalis and Lact. acidophilus may be used to produce fermented goat milk with high counts of both probiotic strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Goat milk fermented with Bif. animalis and Lact. acidophilus can be manufactured as an alternative probiotic dairy product.