| 抑制奶牛乳房炎源金黄色葡萄球菌的乳酸菌的筛选 |
| |
| 引用本文： | 陈宏伟,姜云,郭雪峰,张秀萍.抑制奶牛乳房炎源金黄色葡萄球菌的乳酸菌的筛选[J].中国预防兽医学报,2020(2):128-132. |
| |
| 作者姓名： | 陈宏伟 姜云 郭雪峰 张秀萍 |
| |
| 作者单位： | 塔里木大学动物科学学院;新疆生产建设兵团第二师畜牧兽医工作站 |
| |
| 基金项目： | 塔里木大学校长自然基金项目(TDZKSS201403);新疆生产建设兵团科技创新中青年领军人才项目(2016BC001)。 |
| |
| 摘 要： | 为获得抑制奶牛乳房炎源金黄色葡萄球菌(S. aureus)的乳酸菌(LAB),本研究从新疆巴音布鲁克牧区鲜牛乳和哈萨克族乳制品奶疙瘩样品中分离培养LAB,通过传统的分离鉴定与16S rDNA基因序列测序相结合的方法鉴定LAB种类,同时以临床奶牛乳房炎源金黄色葡萄球菌S. aureus N2为指示菌,采用双层琼脂扩散法检测分离株的抑菌能力。通过测定生长曲线确定分离株的生长稳定期,进而利用硫酸铵沉淀法透析提取稳定期内分离株的细菌素,并检测其细菌素抑菌效价。结果显示:从样品中筛选获得5株能够抑制指示菌生长的LAB,分别为希氏乳杆菌、干酪乳杆菌、粪肠球菌、戊糖片球菌和乳酸乳球菌亚种。生长规律曲线表明20 h^30 h为5株LAB的稳定期,此期培养液pH值维持在3.8~4.5。从培养20 h的5株LAB上清液中提取到了细菌素,经检测其具有抑菌活性,抑菌效价分别为457 IU/mL、1 023 IU/mL、676 IU/mL、1 862 IU/mL和1 023 IU/mL。本研究结果表明5株LAB通过在生长稳定期内维持较低酸性环境(pH<4.5),代谢产生细菌素对乳房炎源S. aureus发挥抑制生长作用。本研究为S. aureus性奶牛乳房炎的生物防治提供了实验依据。
|
| 关 键 词： | 金黄色葡萄球菌 LAB 鉴定 抑菌活性 |
Isolation and characterization lactic acid bacteria against mastitis-causing Staphylococcus aureus |
| |
| CHEN Hong-wei,JIANG Yun,GUO Xue-feng,ZHANG Xiu-ping.Isolation and characterization lactic acid bacteria against mastitis-causing Staphylococcus aureus[J].Chinese Journal of Preventive Veterinary Medicine,2020(2):128-132. |
| |
| Authors: | CHEN Hong-wei JIANG Yun GUO Xue-feng ZHANG Xiu-ping |
| |
| Institution: | (College of Animal Science,Tarim University,Alar 843300,China;Animal Husbandry and VeterinaryWorkstation of the Second Division of Xinjiang Production and Construction Corps,Tiemenguan 841000,China) |
| |
| Abstract: | The objective of this study was to isolate promising lactic acid bacteria(LAB) with antimicrobial activity against Staphylococcus aureus. Firstly, samples of raw milk and traditional Kazakh cheese from Bayanbulak pastoral area of Xinjiang were inoculated on MRS agar for culturing and enrichment of LAB, followed by evaluating the antimicrobial activity of the isolated strains using the double-layer agar diffusion method, choosing Staphylococcus aureus N2 as the indicator bacteria. Secondly, the identification of the potential isolates were further verified on the basis of phenotypic characterization and 16Sr DNA gene sequencing. Finally, growth curves of the strains were created to determine optimal time for harvesting antibacterial metabolites, and then ammonium sulfate precipitation method was carried out for purification of bacteriocin from the isolates. Moreover,antimicrobial titers of purified bacteriocins were also assessed by the double layer agar diffusion method. As a result, five isolates exhibited strong antimicrobial activity against Staphylococcus aureus N2 from the samples, and they were identified as Lactobacillus hilgardii, Lactobacillus casei, Enterococcus faecalis, Pediococcus pentosaceus, and Lactococcus lactis subsp., respectively. Bacterial growth curves indicated that the stationary phases began from 20th hour until 30th hour, and the p H value decreased from 4.5 to 3.8 during this phase. Finally crude bacteriocins were extracted and lyophilized for preservation. All of the bacteriocins demonstrated antimicrobial activity, and their antimicrobial titers were 457 IU/mL, 1 023 IU/mL, 676 IU/mL, 1 862 IU/mL and 1 023 IU/mL,respectively. In conclusion, our study revealed that five isolates displayed antibacterial activity against Staphylococcus aureus N2 by releasing bacteriocins in acidic circumstance(pH<4.5). Considering the inhibitory potential of these strains, their bacteriocins may be useful in controlling bacterial pathogen of bovine mastitis. |
| |
| Keywords: | Staphylococcus aureus lactic acid bacteria(LAB) bacteria strain identification antimicrobial activity |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
