Henri-Dubernet Ségolène, Desmasures Nathalie, Guéguen Micheline
Fromageries BEL S.A, Departement Recherche Appliquee Groupe, 7 bd de l'Industrie, 41100 Vendome, France.
Can J Microbiol. 2008 Mar;54(3):218-28. doi: 10.1139/w07-137.
The diversity and dynamics of Lactobacillus populations in traditional raw milk Camembert cheese were monitored throughout the manufacturing process in 3 dairies. Culture-dependent analysis was carried out on isolates grown on acidified de Man - Rogosa - Sharpe agar and Lactobacillus anaerobic de Man Rogosa Sharpe agar supplemented with vancomycin and bromocresol green media. The isolates were identified by polymerase chain reaction - temperature gradient gel electrophoresis (PCR-TGGE) and (or) species-specific PCR and (or) sequencing, and Lactobacillus paracasei and Lactobacillus plantarum isolates were characterized by pulsed field gel electrophoresis (PFGE). Milk and cheese were subjected to culture-independent analysis by PCR-TGGE. Presumed lactobacilli were detected by plate counts throughout the ripening process. However, molecular analysis of total DNA and DNA of isolates failed to detect Lactobacillus spp. in certain cases. The dominant species in the 3 dairies was L. paracasei. PFGE analysis revealed 21 different profiles among 39 L. paracasei isolates. Lactobacillus plantarum was the second most isolated species, but it occurred nearly exclusively in one dairy. The other species isolated were Lactobacillus parabuchneri, Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus helveticus, a Lactobacillus psittaci/delbrueckii subsp. bulgaricus/gallinarum/crispatus group, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, Lactobacillus brevis, Lactobacillus kefiri, and Lactobacillus perolens. Lactobacilli diversity at the strain level was high. Dynamics varied among dairies, and each cheese exhibited a specific picture of species and strains.
在3家乳制品厂的整个生产过程中,对传统生乳卡门培尔奶酪中乳酸菌种群的多样性和动态变化进行了监测。对在酸化的德氏-罗格斯-夏普琼脂以及添加了万古霉素和溴甲酚绿的乳酸杆菌厌氧德氏-罗格斯-夏普琼脂上生长的分离株进行了基于培养的分析。通过聚合酶链反应-温度梯度凝胶电泳(PCR-TGGE)和(或)种特异性PCR和(或)测序对分离株进行鉴定,并用脉冲场凝胶电泳(PFGE)对副干酪乳杆菌和植物乳杆菌分离株进行特征分析。通过PCR-TGGE对牛奶和奶酪进行非培养分析。在整个成熟过程中通过平板计数检测假定的乳酸菌。然而,在某些情况下,对总DNA和分离株DNA的分子分析未能检测到乳杆菌属。3家乳制品厂中的优势菌种是副干酪乳杆菌。PFGE分析显示,39株副干酪乳杆菌分离株中有21种不同的图谱。植物乳杆菌是第二常见的分离菌种,但几乎只在一家乳制品厂中出现。其他分离出的菌种有布氏副乳杆菌、发酵乳杆菌、嗜酸乳杆菌、瑞士乳杆菌、鹦鹉热/德氏乳杆菌保加利亚亚种/鸡乳杆菌/卷曲乳杆菌组、鼠李糖乳杆菌、德氏乳杆菌保加利亚亚种、德氏乳杆菌乳酸亚种、短乳杆菌、开菲尔乳杆菌和佩罗乳杆菌。菌株水平上的乳酸菌多样性很高
。不同乳制品厂之间的动态变化各不相同,每种奶酪都呈现出特定的菌种和菌株情况。