Corroler D, Mangin I, Desmasures N, Gueguen M
Laboratoire de Microbiologie Alimentaire, Université de Caen, Basse Normandie, 14032 Caen Cedex, France.
Appl Environ Microbiol. 1998 Dec;64(12):4729-35. doi: 10.1128/AEM.64.12.4729-4735.1998.
The genetic diversity of lactococci isolated from raw milk in the Camembert cheese Registered Designation of Origin area was studied. Two seasonal samples (winter and summer) of raw milk were obtained from six farms in two areas (Bessin and Bocage Falaisien) of Normandy. All of the strains analyzed had a Lactococcus lactis subsp. lactis phenotype, whereas the randomly amplified polymorphic DNA (RAPD) technique genotypically identified the strains as members of L. lactis subsp. lactis or L. lactis subsp. cremoris. The genotypes were confirmed by performing standard PCR with primers corresponding to a region of the histidine biosynthesis operon. The geographic distribution of each subspecies of L. lactis was determined; 80% of the Bocage Falaisien strains were members of L. lactis subsp. lactis, and 30.5% of the Bessin strains were members of L. lactis subsp. lactis. A dendrogram was produced from a computer analysis of the RAPD profiles in order to evaluate the diversity of the lactococci below the subspecies level. The coefficient of similarity for 117 of the 139 strains identified as members of L. lactis subsp. cremoris was as high as 66%. The L. lactis subsp. lactis strains were more heterogeneous and formed 10 separate clusters (the level of similarity among the clusters was 18%). Reference strains of L. lactis subsp. lactis fell into 2 of these 10 clusters, demonstrating that lactococcal isolates are clearly different. As determined by the RAPD profiles, some L. lactis subsp. lactis strains were specific to the farms from which they originated and were recovered throughout the year (in both summer and winter). Therefore, the typicality of L. lactis subsp. lactis strains was linked to the farm of origin rather than the area. These findings emphasize the significance of designation of origin and the specificity of "Camembert de Normandie" cheese.
对卡门贝奶酪法定产区生乳中分离出的乳球菌的遗传多样性进行了研究。从诺曼底两个地区(贝辛和博卡日法莱斯)的六个农场采集了两个季节(冬季和夏季)的生乳样本。所有分析的菌株均具有乳酸乳球菌乳酸亚种的表型,而随机扩增多态性DNA(RAPD)技术从基因型上鉴定这些菌株为乳酸乳球菌乳酸亚种或乳酸乳球菌乳脂亚种的成员。通过使用对应于组氨酸生物合成操纵子区域的引物进行标准PCR来确认基因型。确定了乳酸乳球菌各亚种的地理分布;博卡日法莱斯80%的菌株为乳酸乳球菌乳酸亚种的成员,贝辛30.5%的菌株为乳酸乳球菌乳酸亚种的成员。通过对RAPD图谱进行计算机分析生成了一个树形图,以评估亚种水平以下乳球菌的多样性。在鉴定为乳酸乳球菌乳脂亚种成员的139株菌株中,有117株的相似系数高达66%。乳酸乳球菌乳酸亚种的菌株更为异质,形成了10个独立的簇(簇间相似性水平为18%)。乳酸乳球菌乳酸亚种的参考菌株归入这10个簇中的2个,表明分离的乳球菌明显不同。根据RAPD图谱确定,一些乳酸乳球菌乳酸亚种的菌株特定于其来源的农场,并且全年(夏季和冬季)均可分离到。因此,乳酸乳球菌乳酸亚种菌株的典型性与来源农场而非地区有关。这些发现强调了原产地命名的重要性以及“诺曼底卡门贝”奶酪的特异性。