Tuomola E M, Salminen S J
Department of Biochemistry and Food Chemistry, University of Turku, Finland.
Int J Food Microbiol. 1998 May 5;41(1):45-51. doi: 10.1016/s0168-1605(98)00033-6.
The adhesion of 12 different Lactobacillus strains was studied using Caco-2 cell line as an in vitro model for intestinal epithelium. Some of the strains tested have been used as probiotics, and most of them are used in the dairy and food industry. Human and bovine enterotoxigenic Escherichia coli strains were used as positive and negative control, respectively. Bacterial adhesion to Caco-2 cell cultures was quantitated using radiolabelled bacteria. The adherence of bacteria was also observed microscopically after Gram staining. Viability of bacteria prior to adhesion was verified using flow cytometry. Among the tested strains, L. casei (Fyos) was the most adhesive strain and L. casei var. rhamnosus (Lactophilus) was the least adhesive strain, approximately 14 and 3% of the added bacteria adhered to Caco-2 cell cultures, respectively. The corresponding values for positive and negative control E. coli strains were 14 and 4%, respectively. The Lactobacillus strains tested could not be divided into distinctly adhesive or non-adhesive strains, since there was a continuation of adhesion rates. The four most adhesive strains were L. casei (Fyos), L. acidophilus 1 (LC1), L. rhamnosus LC-705 and Lactobacillus GG (ATCC 53103). No significant differences in the percentage adhesion were observed between these strains. Adhesion of all the strains was dependent on the number of bacteria used, since an approximately constant number of Caco-2 cells was used, indicating that the Caco-2 cell binding sites were not saturated. Viability of bacteria was high since approximately 90% of the bacteria were viable with the exception of L. acidophilus 1 which was 74% viable. Microscopic evaluations agreed with the radiolabelled binding as evidenced by observing more bacteria in Gram-stained preparations of good adhering strains compared to poorly adhering strains.
使用Caco - 2细胞系作为肠上皮的体外模型,研究了12种不同乳酸杆菌菌株的黏附情况。所测试的一些菌株已被用作益生菌,其中大多数用于乳制品和食品工业。分别使用人和牛的产肠毒素大肠杆菌菌株作为阳性和阴性对照。使用放射性标记的细菌对细菌与Caco - 2细胞培养物的黏附进行定量。革兰氏染色后,还通过显微镜观察细菌的黏附情况。使用流式细胞术验证黏附前细菌的活力。在所测试的菌株中,干酪乳杆菌(Fyos)是黏附性最强的菌株,而鼠李糖干酪乳杆菌变种(Lactophilus)是黏附性最弱的菌株,分别约有14%和3%添加的细菌黏附到Caco - 2细胞培养物上。阳性和阴性对照大肠杆菌菌株的相应值分别为14%和4%。所测试的乳酸杆菌菌株不能明显分为高黏附性或非黏附性菌株,因为黏附率呈连续变化。黏附性最强的四种菌株是干酪乳杆菌(Fyos)、嗜酸乳杆菌1(LC1)、鼠李糖乳杆菌LC - 705和鼠李糖乳杆菌GG(ATCC 53103)。这些菌株之间在黏附百分比上未观察到显著差异。所有菌株的黏附都取决于所用细菌的数量,因为使用的Caco - 2细胞数量大致恒定,这表明Caco - 2细胞结合位点未饱和。细菌的活力较高,除嗜酸乳杆菌1的活力为74%外,约90%的细菌具有活力。显微镜评估与放射性标记结合情况一致,与黏附性差的菌株相比,在黏附性好的菌株的革兰氏染色制剂中观察到更多细菌即可证明。