健康奶牛子宫颈乳酸菌筛选方法的建立及其抑菌活性分析

doi:10.16431/j.cnki.1671-7236.2018.06.029

Abstract

Abstract:

To select the lactic acid bacteria strains that could be used to prevent and control the metritis in dairy cows,healthy dairy cows 30 to 50 d postpartum were selected to collect the secretions of the cervix,and the lactic acid bacteria were screened by MRS selective medium.The purified strains were subjected to microscopic examination,and the suspected strains with the typical morphology of Lactococcus lactis or Lactobacillus were biochemically identified.The genomic DNA of the isolated strains was extracted,and the 16S rDNA sequence was amplified and sequenced.The sequence of the 16S rDNA sequences was aligned with the NCBI nucleic acid database to identify the genus of the isolates.Antibacterial activity of isolated strains against pathogenic Staphylococcus aureus and Escherichia coli was evaluated using agar diffusion method.The adhesion activity of the isolated strains to goat epithelial cells was also tested.As a result,13 strains of lactic acid bacteria were obtained by staining microscopic examination,biochemical identification and 16S rDNA sequencing.Among them,5 strains had strong antibacterial activity,6 strains had strong ability of adhering epithelial cells,strains of No.7,No.12 and No.23 both had good antibacterial activity and adhesion of epithelial cells.In summary,the experiment established a selected method for probiotic lactic acid bacteria in healthy cow's cervix.3 strains of lactic acid bacteria with good probiotic properties were obtained,which provided the basic materials for the preparation of probiotics and was of great significance for the prevention and treatment of dairy cows.

Key words: dairy cow metritis; lactic acid bacteria; probiotics

CLC Number:

S858.23

SANG Mengqi, YU Qin, SHAO Dan, DONG Shuwei, WANG Dongsheng, ZHANG Shidong, WU Xiaohu, YAN Zuoting. Establishment of Selected Method for Lactic Acid Bacteria in Healthy Cows and Analysis of Its Antibacterial Activity[J]. , 2018, 45(6): 1653-1660.

References

[1] 严作廷,王东升,张世栋,等.奶牛子宫内膜炎的防治[J].中国畜牧兽医文摘,2011,27(6):97-98. YAN Z T,WANG D S,ZHANG S D,et al.Prevention and treatment of cow endometritis[J].Chinese Abstracts of Animal Husbandry and Veterinary Medicine,2011,27(6):97-98.(in Chinese)
[2] 王玉舜,戚建允.百福他和美达佳联合用药治疗奶牛子宫炎疗效研究[J].中国奶牛,2015,2:24-25. WANG Y S,QI J Y.Therapeutic effect of combination therapy with Bio-Mycin and Metacam on cows with metritis[J].China Dairy Cattle,2015,2:24-25.(in Chinese)
[3] 魏利杰,徐琳,王锦卓.乳酸杆菌代谢产物对阴道常见致病菌抑制作用的试验研究[J].吉林医学,2013,34(13):2475-2476. WEI L J,XU L,WANG J Z.Experimental study on the inhibitory effect of metabolites of Lactobacillus on common vaginal pathogens[J].Jilin Medical Journal,2013,34(13):2475-2476.(in Chinese)
[4] 刘军,宁安红,张蓓蓓,等.乳酸杆菌代谢产物对一些常见菌黏附阴道上皮细胞的抑制作用的初步探讨[J].中国微生态学杂志,2009,21(8):705-706. LIU J,NING A H,ZHANG B B,et al.Preliminary study on the inhibition of Lactobacillus metabolites to some common bacteria adhesion of vaginal epithelial cells[J].Chinese Journal of Microbiology,2009,21(8):705-706.(in Chinese)
[5] 吴惠芬,毛胜勇,姚文,等.猪源乳酸菌的分离及其发酵产物的比较[A].全国饲料营养学术研讨会论文集[C].2002. WU H F,MAO S Y,YAO W,et al.Isolation of lactic acid bacteria from pig and the comparison of its fermentation products[A].National Feeding Nutrition Symposium[C].2002.(in Chinese)
[6] WANG J,SUN C,LIU C,et al.Comparison of vaginal microbial community structure in healthy and endometritis dairy cows by PCR-DGGE and Real-time PCR[J].Anaerobe,2016,38:1-6.
[7] SAMOT J,LEBRETON J,BADET C.Adherence capacities of oral lactobacilli for potential probiotic purposes[J].Anaerobe,2011,17(2):69-72.
[8] 卢良恕.中国大麦学[M].北京:中国农业出版社,1996. LU L S.Chinese Barley Science[M].Beijing:China Agriculture Press,1996.(in Chinese)
[9] 马静,朱惠莲,唐志红,等.乳酸菌对人体肠道黄曲霉毒素排泄的影响[J].现代食品科技,2003,19(s1):23-26. MA J,ZHU H L,TANG Z H,et al.Effect of lactic acid bacteria on human intestinal aflatoxin excre-tion[J].Modern Food Science & Technology,2003,19(s1):23-26.(in Chinese)
[10] 沈定树,陈倩.乳酸菌对人体益处的研究进展[J].中国卫生检验杂志,2012,22(5):1208-1210. SHEN D S,CHEN Q.Research progress on the benefits of lactic acid bacteria to human body[J].China Journal of Health Laboratory Technology,2012,22(5):1208-1210.(in Chinese)
[11] RD C J.Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases[J].Clinical Microbiology Reviews,2004,17(4):840-862.
[12] 杨玲玲,职晓阳,李文均.拟诺卡氏菌16S rRNA,gyrB,sod和rpoB基因的系统发育分析[J].微生物学报,2007,47(6):951-955. YANG L L,ZHI X Y,LI W J.Phylogenetic analysis of Nocardiopsis species based on 16S rRNA,gyrB, sod and rpoB gene sequences[J].Acta Microbiologica Sinica,2007,47(6):951-955(in Chinese)
[13] 薛洋洋.微生态制剂治疗奶牛子宫内膜炎的试验研究[D].大庆:黑龙江八一农垦大学,2014. XUE Y Y.Probiotics experimental study for the treatment of cow endometritis[D].Daqing:Heilongjiang Bayi Agricultural University,2014.(in Chinese)
[14] 姚火春.兽医微生物学实验指导[M].北京:中国农业出版社,2002. YAO H C.Veterinary Microbiology Experimental Guide[M].Beijing:China Agriculture Press,2002.(in Chinese)
[15] 刘冬梅,李理,杨晓泉,等.用牛津杯法测定益生菌的抑菌活力[J].食品研究与开发,2006,27(3):110-111. LIU D M,LI L,YANG X Q,et al.Determination of antibacterial activity of probiotics by oxford plate assay system[J].Food Research and Development,2006,27(3):110-111.(in Chinese)
[16] VINDEROLA C G,MOCCHIUTTI P,REINHEIMER J A.Interactions among lactic acid starter and probiotic bacteria used for fermented dairy products[J].Journal of Dairy Science,2002,85(4):721-729.
[17] 李向楠.奶牛生殖道内益生乳酸菌的分离、筛选及功能特性研究[D].长春:吉林农业大学,2014. LI X N.Isolation,screening and study of the functional characteristics of probiotics from the cow genital tract[J].Changchun:Jilin Agricultural University,2014.(in Chinese)
[18] 王孝武,王旭荣,杨志强,等.奶牛子宫内膜炎研究进展[J].动物医学进展,2014,35(7):98-102. WANG X W,WANG X R,YANG Z Q,et al.Research progress on cow endometritis[J].Progress in Animal Medicine,2014,35(7):98-102.(in Chinese)
[19] 王红艳,贾继辉,邢咏梅, 等.乳酸杆菌与阴道微生态关系的研究进展[J].潍坊医学院学报, 2004,26(2):145-147. WANG H Y,JIA J H,XING Y M,et al.Progress in research on the relationship between lactobacilli and vaginal microflora[J].Acta Academiae Medicine Weifang,2004,26(2):145-147.(in Chinese)
[20] 王凡,王建设,刘婷,等.一株猪源屎肠球菌的分离、鉴定及益生功能初步研究[J].中国畜牧兽医,2012,39(7):231-234. WANG F,WANG J S,LIU T,et al.Isolation,identification of Enterococcus faecium strain and preliminary study on its beneficial effects[J].China Animal Husbandry & Veterinary Medicine,2012,39(7):231-234.
[21] PERSING D H,TENOVER F C,TANG Y W,et al.Molecular Microbiology:Diagnostic Principles and Practice[M].Washington DC:Asm Press,2016.
[22] 于晨龙,张七斤,陈光明,等.不同来源乳酸菌的分离与鉴定[J].中国畜牧兽医,2014,41(2):208-212. YU C L,ZHANG Q J,CHEN G M,et al.Isolation and identification of Lactobacillus from different origins[J].China Animal Husbandry & Veterinary Medicine,2014,41(2):208-212.(in Chinese)
[23] 杨龙龙,蒋建军,剡根强,等.牛瘤胃乳酸菌的分离与鉴定[J].中国畜牧兽医,2014,41(8):145-148. YANG L L,JANG J J,YAN G Q,et al.Isolation and identification of lactic acid bacteria from bovine rumen[J].China Animal Husbandry & Veterinary Medicine,2014,41(8):145-148.(in Chinese)
[24] REID G,MC GROARTY J A,ANGOTTI R,et al.Lactobacillus inhibitor production against Escherichia coli and coaggregation ability with uropathogens[J].Journal of Microbiology,1985,34(4):344-351.
[25] 王秋菊,崔一喆,刘胜军,等.笼养成年蛋鸡盲肠中乳酸菌的分离与鉴定[J].中国畜牧兽医,2013,40(8):112-115. WANG Q J,CUI Y Z,LIU S J,et al.Isolation and identification of lactoc acid bacteria in cecum of laying hens fed in cage[J].China Animal Husbandry & Veterinary Medicine,2013,40(8):112-115.(in Chinese)
[26] 韩秋霞,李春露,崔志芳.体外条件下肉鸡源乳酸菌抑菌作用研究[J].中国畜牧兽医,2006,33(3):65-67. HAN Q X,LI C L,CUI Z F.Research on the in-vitro bacteriostatic effect of Lactobacillus from the intestine of chicken[J].China Animal Husbandry & Veterinary Medicine,2006,33(3):65-67.(in Chinese)
[27] 田丰松,王军,杨连玉,等.不同乳酸菌分离株抗菌物质及其抗菌活性的体外检测及鉴定[J].中国预防兽医学报,2016,38(7):558-561. TIAN F S,WANG J,YANG L Y,et al.In vitro analysis of substance with antibacterial activity of lactic acid bacteria[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(7):558-561.(in Chinese)
[28] 许女,史改玲,程宏乾,等.牛源乳酸菌的粘附特性及对奶牛乳房炎大肠杆菌的粘附抑制研究[J].中国食品学报,2017,17(1):20-30. XU N,SHI G L,CHENG H Q,et al.Adhesive properties of lactic acid bacteria derived from bovine larvae and their inhibitory effects on the adhesion of Escherichia coli in mastitis in dairy cows[J].Journal of Chinese Institute of Food Science and Technology,2017,17(1):20-30.(in Chinese)
[29] SANDRA G,ALEX B,FRANCESC F,et al.Protential of lactic acid bacteria at regulating Escherichia coli infection and inflammation of bovine endome-trium[J].Theriogenology,2016,85(4):625-637.

Establishment of Selected Method for Lactic Acid Bacteria in Healthy Cows and Analysis of Its Antibacterial Activity

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