三种益生菌菌株与PK-15细胞共培养抗TGEV作用

为模拟猪肠道内环境,探究益生菌与细胞共培养抗TGEV作用,利用Transwell小室在体外建立益生菌与PK-15细胞共培养,通过台盼蓝染色,检测PK-15细胞存活率。结果表明,当益生菌菌体浓度102cfu·m L-1≤c≤1012cfu·m L-1,共培养时间为1和3 h;当共培养时间1 h≤t≤24 h,益生菌菌体浓度为102、104、106和108cfu·m L-1时,PK-15细胞存活率较高,益生菌与PK-15细胞共培养成立。采用MTT比色法检测对TGEV抑制率,结果显示,益生菌菌体浓度约为102~108cfu·m L-1,共培养时间为1~24 h时对TGEV抑制率均达90%以上。此外,益生菌与PK-15细胞共培养的TGEV平均滴度较低。

Anti-TGEV effect of three probiotics strains and PK-15 cells co-culture

In order to simulate the porcine intestinal environment and explore the anti-TGEV effect of probiotics and cel s co-culture, this experiment used Transwel to establish probiotics and PK-15 cel s co-culture in vitro. In addition, this test took advantage of trypan blue staining to detect the PK-15 cel s survival rates. The results showed that the PK-15 cel s survival rates were higher under the conditions of 102 cfu·mL-1≤the concentration of probiotics≤1012 cfu·mL-1, 1, 3 h of co-culture time or 1 h≤the time of co-culture≤24 h, 102, 104, 106 and 108 cfu·mL-1 of probiotics concentration. On this basis, this experiment made use of MTT colorimetric assay to detect the TGEV inhibition rates. The results showed that the TGEV inhibition rates were more than 90%when the concentration of probiotics was about 102-108 cfu·mL-1 and the time of co-culture was about 1-24 h. Furthermore, the TGEV average titer of probiotics and PK-15 cel s co-culture was lower.