文章摘要
抑制猪霍乱沙门氏菌的乳酸菌的分离、鉴定及潜在抑菌物质分析
Isolation and Identification of Lacto-Bacteria Inhibiting Salmonella choleraesuis and Analysis of Its Potential Bacteriostatic Substance
  
DOI:doi:10.3969/j.issn.1005-7021.2020.04.008
中文关键词: 乳酸菌  猪霍乱沙门氏菌  抑菌活性  抑菌物质
英文关键词: lacto-bacteria  Salmonella choleraesuis  inhibitory activity  bacteriostatic component
基金项目:国家自然科学基金项目(31760042,31960286)
作者单位
李宏伟 昆明理工大学 生命科学与技术学院云南 昆明 650500 
肖瑶 昆明理工大学 生命科学与技术学院云南 昆明 650500 
张关令 昆明理工大学 生命科学与技术学院云南 昆明 650500 
林连兵 昆明理工大学 生命科学与技术学院云南 昆明 650500 
张棋麟 昆明理工大学 生命科学与技术学院云南 昆明 650500 
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中文摘要:
      猪霍乱沙门氏菌(Salmonella choleraesuis)是一种常见的食源性致病菌。为了预防和治疗该菌引起的疾病,饲养者在饲料中大量添加抗生素,致使猪肉存在严重的食品安全问题。从健康成年无量山乌骨鸡肠道内容物中筛选出具有抑菌作用的乳酸菌,测定其对猪霍乱沙门氏菌的抑菌效果,分析乳酸菌抑制猪霍乱沙门氏菌的有效成分,并对筛选的乳酸菌种进行了分子生物学鉴定。采用双层平板法对具有抑制猪霍乱沙门氏菌的乳酸菌进行筛选,采用牛津杯法对抑菌效果进行测定,并采用酶蛋白敏感性测定、热处理、有机酸处理等方法分析抑菌活性物质有效成分,采用16S rDNA分子标记对乳酸菌进行鉴定,并构建系统发育树。结果显示,从健康鸡肠道中筛选出18株乳酸菌,其中2株对猪霍乱沙门氏菌(Salmonella choleraesuis)、大肠埃希菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)、肠炎沙门氏菌(Salmonella enteritidis)、肠炎沙门氏菌亚种(Salmonella enteritidis subspecies)、志贺氏菌(Shigella)、无乳链球菌(Streptococcus agatactiae)具有良好的抑菌效果;不同蛋白酶、pH处理对乳酸菌无细胞培养液抑菌效果均有不同程度的影响,但是经80 ℃处理的乳酸菌无细胞培养液,其抑菌效果未明显改变。经鉴定,2株乳酸菌分别为植物乳杆菌(Lactobacillus plantarum)和短乳杆菌(Lactobacillus brevis)。从健康成年无量山乌骨鸡肠道内容物中分离得到的植物乳杆菌菌株L2和短乳杆菌菌株L4对猪霍乱沙门氏菌等致病菌具有明显地抑制作用,推测其抑菌有效成分可能为小肽类及有机酸,这对减少抗生素使用,提高猪肉食品的品质与安全性具有一定价值。
英文摘要:
      Salmonella choleraesuis is a common food-borne pathogen. In order to prevent and treat the diseases caused by the bacterium, a large amount of antibiotics was added into the feed, causing serious food safety and quality problems. Lacto-bacteria with antibacterial activity were screened out from intestinal contents of adult healthy Wuliangshan black-bone chicken, their antibacterial effects on S. choleraesuis were determined and analyzed for effective components of the lacto bacteria against S. choleraesuis, and the lacto-bacteria strain obtained in this study were identified using molecular biology. The lacto-bacteria that can inhibit S. choleraesuis were screened by the double-layer plate method. The antibacterial effect was determined by the Oxford Cup method. The effective components of antibacterial active substances were analyzed by the enzyme protein sensitivity experiments, heat and organic acid treatments. The lacto-bacteria were determined using 16S rDNA molecular marker, and a phylogenetic tree was reconstructed. The results showed that 18 strains of lacto-bacteria screened out in this study, two of them presented significant antibacterial effects on S. choleraesuis, Escherichia coli, Staphylococcus aureus, Salmonella enteritidis, Salmonella enteritidis subspeies, Shigella and Streptococcus agatactiae; different proteases and pH treatment showed different degrees of effects on antibacterial effects of the fermented broth of lacto-bacteria, but antibacterial effects of the lacto bacteria fermented broth has no significant changes after treated at 80 ℃. The two strains of lacto-bacteria were identified as Lactobacillus plantarum and Lactobacillus brevis using 16S rDNA. L. plantarum strain L2 and L. brevis strain L4 isolated from the intestinal contents of adult healthy Wuliangshan black-bone chicken have significant inhibitory effects on pathogen, S. choleraesuis etc, and it is speculated that small peptides and organic acids may be potential antibacterial components of these two Lactobacillus strains, having a certain value for reducing the use of antibiotics to improve of the quality and security of pork-derived food.
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