Cronin T, Ziino M, Condurso C, McSweeney P L H, Mills S, Ross R P, Stanton C
Teagasc, Biotechnology Centre, Moorepark, Fermoy, County Cork, Ireland.
J Appl Microbiol. 2007 Oct;103(4):1128-39. doi: 10.1111/j.1365-2672.2007.03364.x.
The microbial and chemical composition of seven different semi-ripened (45 days) Provola dei Nebrodi Sicilian cheese samples were assessed in order to investigate the diversity of the microbial population in cheese made from different geographical areas throughout Sicily.
The samples, which were obtained from seven different Provola dei Nebrodi manufacturers, were assessed using selective media. Interestingly, concentrations of presumptive lactobacilli represented over 90% of the total microbial population. In total, 105 presumptive Lactobacillus isolates were characterized to determine the relatedness of the isolates between the seven different cheeses. Randomly amplified polymorphic DNA polymerase chain reaction (RAPD PCR) analysis of the 105 presumptive lactobacilli indicated the presence of 22 distinct isolates. Further investigation of the isolates using pulsed field gel electrophoresis (PFGE) following restriction with the enzyme ApaI revealed the presence of 19 distinct macrorestriction patterns and the presence of between one and four distinct isolates per cheese sample (out of a total of 15 isolates per cheese randomly taken from Lactobacillus selective media plates). Analysis of the 16S rDNA sequence of each genetically distinct isolate demonstrated the dominance of the Lactobacillus casei species in all cheese samples assessed. Lactobacillus delbrueckii and Pediococcus pentosaceus species were also detected. The concentration of free amino acids, used to estimate the extent of proteolysis in each cheese, ranged from 59 to 433 mg 100 g(-1) cheese.
Microbiological assessment of the cheeses demonstrated the dominance of Lactobacillus species after 45 days of ripening with levels ranging from 8.3 to 9.4 log CFU g(-1).
This study provides new information on the diversity of lactobacilli within an artisanal Sicilian cheese, enabling the identification of 17 strains of Lact. casei, one strain of Lact. delbrueckii and Ped. pentosaceus through the combined use of RAPD PCR, PFGE and 16S rDNA sequencing.
对七个不同的半熟(45天)西西里内布罗迪普罗沃拉奶酪样品的微生物和化学成分进行评估,以调查整个西西里岛不同地理区域生产的奶酪中微生物种群的多样性。
从七个不同的西西里内布罗迪普罗沃拉奶酪制造商处获取样品,使用选择性培养基进行评估。有趣的是,推测的乳酸杆菌浓度占微生物总数的90%以上。总共对105株推测的乳酸杆菌分离株进行了特征分析,以确定七个不同奶酪之间分离株的相关性。对105株推测的乳酸杆菌进行随机扩增多态性DNA聚合酶链反应(RAPD PCR)分析,结果表明存在22种不同的分离株。在用ApaI酶进行限制性内切后,使用脉冲场凝胶电泳(PFGE)对分离株进行进一步研究,结果显示存在19种不同的宏观限制性图谱,每个奶酪样品(从乳酸杆菌选择性培养基平板中随机选取的每个奶酪共15株分离株)中存在1至4种不同的分离株。对每个基因不同的分离株进行16S rDNA序列分析表明,在所有评估的奶酪样品中,干酪乳杆菌占主导地位。还检测到德氏乳杆菌和戊糖片球菌。用于估计每种奶酪中蛋白水解程度的游离氨基酸浓度范围为59至433 mg 100 g(-1)奶酪。
奶酪的微生物学评估表明,成熟45天后乳酸杆菌占主导地位,水平范围为8.3至9.4 log CFU g(-1)。
本研究提供了关于西西里手工奶酪中乳酸杆菌多样性的新信息,通过联合使用RAPD PCR、PFGE和16S rDNA测序,能够鉴定出17株干酪乳杆菌、1株德氏乳杆菌和戊糖片球菌。