牛瘤胃植物乳杆菌的分离鉴定与生物学特性分析
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摘要
牛瘤胃中栖息着复杂多样的微生物,这些微生物种群一般会受饲粮、宿主基因型、健康状况、生活环境以及环境中的共生微生物之间的群体效应等的影响,进而又影响着饲料的利用率和牛群的健康,因此弄清牛瘤胃微生物群是十分必要的。利用厌养分离技术从成年牛瘤胃中分离出一株细菌,命名为Y-A-1。结合细菌形态学、生理生化特性和16Sr RNA序列同源性分析鉴定,结果表明Y-A-1菌株为植物乳杆菌。对该菌的生长特性包括最适生长温度、生长曲线、耐酸能力、耐盐等进行测定,结果表明菌株在30℃,pH 5.5~7.5范围内,盐离子浓度在7.5以下的环境中生长最好,该菌株的分离和生物特性研究对牛瘤胃微生物生存状态有了初步的了解,为进一步了解牛瘤胃微生物群奠定了一定的基础。
There are many microorganisms in bovine rumen,these microbial populations are generally affected by fodder,host genotype,health status,living environment and symbiotic microbes in the environment,they affect the utilization of feed and cattle health,so understanding of bovine rumen microbes is very necessary. In this paper,a bacteria strain was isolated from adult bovine rumen using anaerobic separation technology,and named Y-A-1. Combination of bacterial morphological,physiological and biochemical characteristics and 16 Sr RNA sequence homology analysis,the results showed that the Y-A-1 strain was plant lactobacillus. The growth characteristics of the bacterium were detected including the optimum growth temperature,growth curve,acid-resistant ability and salt tolerance,the results showed that the growth of the bacterium was the best at 30 ℃,between pH 5.5 and pH 7.5 and concentration of salt ion in 7.5. The separation and biological characteristics research of bovine rumen microbial laid a certain foundation in order to further understand bovine rumen microbes.
There are many microorganisms in bovine rumen,these microbial populations are generally affected by fodder,host genotype,health status,living environment and symbiotic microbes in the environment,they affect the utilization of feed and cattle health,so understanding of bovine rumen microbes is very necessary. In this paper,a bacteria strain was isolated from adult bovine rumen using anaerobic separation technology,and named Y-A-1. Combination of bacterial morphological,physiological and biochemical characteristics and 16 Sr RNA sequence homology analysis,the results showed that the Y-A-1 strain was plant lactobacillus. The growth characteristics of the bacterium were detected including the optimum growth temperature,growth curve,acid-resistant ability and salt tolerance,the results showed that the growth of the bacterium was the best at 30 ℃,between pH 5.5 and pH 7.5 and concentration of salt ion in 7.5. The separation and biological characteristics research of bovine rumen microbial laid a certain foundation in order to further understand bovine rumen microbes.
引文
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[26]Lai C H,Wu S R,Pang J C,et al.Designing primers and evaluation of the efficiency of propidium monoazideQuantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius[J].J Food Drug Anal,2017,25(3):533-542.
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[28]Yang Z Q,Xue Y,Rao S Q,et al.Isolation of Probiotic Piliated Lactobacillus rhamnosus Strains from Human Fecal Microbiota Using Spa A Antiserum-Based Colony Immunoblotting[J].J Microbiol Biotechnol,2017,27(11):1971-1982.
[29]Di Marzio M,Rusconi B,Yennawar N H,et al.Identification of a mouse Lactobacillus johnsonii strain with deconjugase activity against the FXR antagonist T-β-MCA[J].PLo S One,2017,12(9):183564.
[2]苏少锋,刘永志,刘红葵,等.瘤胃表皮葡萄球菌的分离鉴定及其β-葡聚糖酶基因的克隆和在乳酸球菌中表达的研究[J].畜牧与饲料科学,2013,34(5):1-6.
[3]卢玉飞,周凌云,赵圣国,等.近10年瘤胃微生物分离培养研究进展[J].中国微生态学杂志,2012,24(9):856-861.
[4]杨承剑,杨炳壮,梁辛,等.水牛营养研究新进展[J].中国畜牧兽医,2013,40(12):85-88.
[5]马晨.奶牛乳房炎与肠道菌群的关联研究[D].呼和浩特:内蒙古农业大学,2016.
[6]马晨,张和平,刘彩虹,等.牛瘤胃与肠道微生物多样性的研究进展[J].动物营养学报,2014(10):66-71.
[7]刘岩岩,张鹭,张春艳,等.一株乳酸菌培养条件优化及菌种鉴定[J].齐齐哈尔大学学报,2008(2):21-24.
[8]代永刚,田志刚,南喜平.乳酸菌及其生理功能研究的进展[J].农产品加工(学刊),2009(7):24-26,29.
[9]金玉铭.乳酸菌发酵及酸奶工艺优化研究[J].民营科技,2014(1):80.
[10]王磊,陈宇飞,刘长姣.发酵饮料的开发现状及研究前景[J].食品工业科技,2015(10):379-382.
[11]赵红霞,詹勇,许梓荣.乳酸菌的研究及其应用[J].江西饲料,2003(1):9-12.
[12]张江巍.乳酸菌抗氧化活性的研究[D].南昌:南昌大学,2006.
[13]高书锋.黄瓜灰霉病和炭疽病生物防治研究[D].郑州:河南农业大学,2006.
[14]赵谦.抗氧化乳杆菌的筛选及其抗氧化活性的研究[D].保定:河北农业大学,2010.
[15]张俊娟.具有黏附性乳杆菌的筛选与特性研究[D].保定:河北农业大学,2011.
[16]杨菲菲,麻丽坤,王增敏.断奶仔猪饲料中乳酸菌的应用[J].上海畜牧兽医通讯,2006(5):60-61.
[17]Goldstein E J,Tyrrell K L,Citron D M.Lactobacillus species:taxonomic complexity and controversial susceptibilities[J].Clin Infect Dis,2015,60(2):98.
[18]Cousin F J,Lynch S M,Harris H M,et al.Detection and genomic characterization of motility in Lactobacillus curvatus:confirmation of motility in a species outside the Lactobacillus salivarius clade[J].Appl Environ Microbiol,2015,81(4):1297-1308.
[19]Yakovlieva M,Tacheva T,Mihaylova S,et al.Influence of Lactobacillus brevis 15 and Lactobacillus plantarum 13on blood glucose and body weight in rats after high-fructose diet[J].Benef Microbes,2015(4):505-512.
[20]Bai fen S,Long zhu J,Yi jing L,et al.Chromosomal Insertions in the Lactobacillus casei upp That Are Useful for Vaccine Expression Gene[J].Appl Environ Microbiol,2014,80(11):3321.
[21]王晓丽,王永山,诸玉梅,等.5株乳酸菌的分离鉴定与生物学特性研究[J].江苏农业科学,2011(1):390-392.
[22]Zagorec M,Champomier-Vergès MC.Lactobacillus sakei:A Starter for Sausage Fermentation,a Protective Culture for Meat Products[J].Microorganisms,2017(3):56.
[23]Tohno M,Tanizawa Y,Irisawa T,et al.Lactobacillus silagincola sp.nov.and Lactobacillus pentosiphilus sp.nov.,isolated from silage[J].Int J Syst Evol Microbiol,2017,67(9):3639-3644.
[24]Hasannejad Bibalan M,Eshaghi M,Rohani M,et al.Isolates of Lactobacillus plantarum and L.reuteri display greater antiproliferative and antipathogenic activity than other Lactobacillus isolates[J].J Med Microbiol,2017,66(10):1416-1420.
[25]Wang G,Li X,Zhao J,et al.Correction:Lactobacillus casei CCFM419 attenuates type 2 diabetes via a gut microbiota dependent mechanism[J].Food Funct,2017,8(10):3814.
[26]Lai C H,Wu S R,Pang J C,et al.Designing primers and evaluation of the efficiency of propidium monoazideQuantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius[J].J Food Drug Anal,2017,25(3):533-542.
[27]Silva FKP,Brück DW,Brück WM.Isolation of proteolytic bacteria from mealworm(Tenebrio molitor)exoskeletons to produce chitinous material[J].FEMS Microbiol Lett,2017,364(17):15.
[28]Yang Z Q,Xue Y,Rao S Q,et al.Isolation of Probiotic Piliated Lactobacillus rhamnosus Strains from Human Fecal Microbiota Using Spa A Antiserum-Based Colony Immunoblotting[J].J Microbiol Biotechnol,2017,27(11):1971-1982.
[29]Di Marzio M,Rusconi B,Yennawar N H,et al.Identification of a mouse Lactobacillus johnsonii strain with deconjugase activity against the FXR antagonist T-β-MCA[J].PLo S One,2017,12(9):183564.