Department of Public Health, Comparative Pathology, and Veterinary, Padova, Italy.
Res Microbiol. 2011 Apr;162(3):231-9. doi: 10.1016/j.resmic.2011.01.002. Epub 2011 Jan 22.
The microbial community in milk is of great importance in the manufacture of traditional cheeses produced using raw milk and natural cultures. During milk curdling and cheese ripening, complex interactions occur in the microbial community, and accurate identification of the microorganisms involved provides essential information for understanding their role in these processes and in flavor production. Recent improvements in molecular biological methods have led to their application to food matrices, and thereby opened new perspectives for the study of microbial communities in fermented foods. In this study, a description of microbial community composition during the manufacture and ripening of Montasio cheese was provided. A combined approach using culture-dependent and -independent methods was applied. Culture-dependent identification was compared with 16S clone libraries sequencing data obtained from both DNA and reverse-transcribed RNA (cDNA) amplification and real-time quantitative PCR (qPCR) assays developed to detect and quantify specific bacterial species/genera (Streptococcus thermophilus, Lactobacillus casei, Pediococcus pentosaceus, Enterococcus spp., Pseudomonas spp.). S. thermophilus was the predominant LAB species throughout the entire ripening period of Montasio cheese. The culture-independent method demonstrates the relevant presence of Pseudomonas spp. and Lactococcus piscium at the beginning of ripening. The culture-dependent approach and the two culture-independent approaches produced complementary information, together generating a general view of cheese microbial ecology.
在使用生奶和天然培养物生产传统奶酪的过程中,牛奶中的微生物群落具有重要意义。在牛奶凝结和奶酪成熟过程中,微生物群落中会发生复杂的相互作用,准确识别涉及的微生物为了解它们在这些过程和风味产生中的作用提供了重要信息。近年来,分子生物学方法的改进使其能够应用于食品基质,从而为发酵食品中微生物群落的研究开辟了新的视角。本研究描述了蒙塔萨罗奶酪在生产和成熟过程中的微生物群落组成。采用了结合依赖培养和非依赖培养方法的方法。与从 DNA 和反转录 RNA(cDNA)扩增以及开发的实时定量 PCR(qPCR)检测和定量特定细菌物种/属(嗜热链球菌、干酪乳杆菌、戊糖片球菌、肠球菌属、假单胞菌属)获得的 16S 克隆文库测序数据相比,进行了依赖培养的鉴定。在蒙塔萨罗奶酪成熟的整个过程中,嗜热链球菌都是主要的 LAB 物种。非依赖培养方法证明了假单胞菌属和乳球菌属在成熟初期的存在。依赖培养方法和两种非依赖培养方法提供了互补信息,共同生成了奶酪微生物生态学的总体视图。