| 摘要: |
| 【目的】从健康仔猪的粪便及各肠段内容物和黏液中分离乳酸菌，并以永生化猪小肠上皮细胞ZYM-SIEC02为体外细胞模型对猪源益生性乳酸菌进行筛选。【方法】采用分离培养和常规生化试验对乳酸菌进行分离与鉴定。用永生化猪小肠上皮细胞ZYM-SIEC02作为体外模型，通过黏附性试验、耐酸耐胆盐试验和对致病菌的黏附抑制试验对菌株进行筛选。【结果】分离筛选出了12株乳酸菌，其黏附能力存在差异。Y091231、Y091221、Y224031、X302032、Y223052和Y223072等6株菌的黏附性较好，黏附指数依次为：2 903.25±83.33，2 320.37±81.02，1 965.43±37.72，1 416.12±62.74，1 100. 00±68.10，988.73±17.06。对其进行耐酸耐胆盐试验和抑菌试验，结果表明，菌株Y091231、Y091221和Y224031对致病菌均具有较强的黏附抑制能力，且对高胆盐、低pH具有较强的耐受性。其中Y091231对大肠杆菌C83921的黏附抑制率可达(99.40±3.17)%，Y091221对致病性沙门氏菌的黏附抑制率为(97.10±2.04)%，Y091231对金黄色葡萄球菌CVCC2258的黏附抑制率为(98.00±2.31)%。Y091231、Y091221和Y224031 3株菌分别被鉴定为发酵乳杆菌、乳酸乳杆菌和嗜酸乳杆菌。【结论】筛选得到的3株益生性乳酸菌，不仅具有耐酸耐胆盐能力，而且对小肠黏膜上皮细胞具有强黏附性，对大肠杆菌C83921、致病性沙门氏菌和金黄色葡萄球菌CVCC2258具有较强的黏附抑制能力，可作为肠道微生态制剂的候选菌株。 |
| 关键词: 永生化猪小肠上皮细胞 猪源乳酸菌 黏附性 黏附抑制 |
| DOI： |
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| 基金项目:西北农林科技大学青年学术骨干教师支持计划项目（Z111020902） |
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| Screening tests of high adhesion Lactobacillus strains isolated from swine based on Immortalization Neonatal Swine Intestinal Epithelial Cells ZYM-SIEC02 |
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| Abstract: |
| 【Objective】The study isolated the probiotics strains from piglets’ manure and contents from different parts of bowel.The Immortalization Neonatal Swine Intestinal Epithelial Cells ZYM-SIEC02 (abbreviation:SIEC) were taken as the model for screening tests.【Method】Isolation and identification of probiotics were performed by isolating culture and conventional physiology biochemistry experiment.We took the Immortalization Neonatal Swine Intestinal Epithelial Cells ZYM SIEC02(abbreviation:SIEC) for the model,screened the strains through the test of resistance capability to bile salts and low pH,adhesion capability to the SIEC and inhibitory effects on the growth of pathogenic bacteria for probiotics filtering.【Result】The research screened 12 strains and the different strains had different adherence abilities(P<0.05).The adhesion index of the“Y091231”,“Y091221”,“Y224031”,“X302032”,“Y223052” and “Y223072” respectively was:2 903.25±83.33,2 320.37±81.02,1 965.43±37.72,1 416.12±62.74,1 100.00±68.10,988.73±17.06.Through testing the tolerability and inhibition rate of the six clones,we got three better ones:“Y091231”,“Y091221”,“Y224031”.And the inhibition rate of “Y091231” against Escherichia coli C83921 was (99.40±3.17)%,similarly,the inhibition rate of “Y091221” against Enteritis salmonella reached (97.10±2.04)% and that of “Y091231” against Staphylococcus CVCC2258 was (98.00±2.31)%.The strains “Y091231”,“Y091221” and “Y224031” were refered as Lactobacillus fermentation,Lactobacillus lactis and Lactobacillus acidophilus respectively.【Conclusion】Lactobacillus from the experiment not only have the capability of resistance to bile salts and low pH,but also can adhere to the SIEC and have inhibitory effects on the growth of E.coli C83921,Enteristis salmonella and Staphylococcus CVCC2258).So they may do for the intestinal tract micro ecology preparation candidate strains. |
| Key words: Immortalization Neonatal Swine Intestinal Epithelial Cells Lactobacillus of piglets intestines adhesion adhesion inhibition |
