| 长白山原始森林土壤源耐低温乳酸菌的筛选 |
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| 引用本文： | 陈龙,魏炳栋,郑琳,张莹,闫晓刚,于维.长白山原始森林土壤源耐低温乳酸菌的筛选[J].中国畜牧兽医,2020,47(10):3103-3113. |
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| 作者姓名： | 陈龙 魏炳栋 郑琳 张莹 闫晓刚 于维 |
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| 作者单位： | 吉林省农业科学院畜牧科学分院, 公主岭 136100 |
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| 基金项目： | 吉林省农业科技创新工程博士后基金项目（C92072030）；吉林省农业科技创新工程引进博士及高层次人才科研启动基金项目（C92072032）；吉林省科研院所引进高层次科技创新人才资助计划（2018001）；吉林省农业科技创新工程金项目（CXGC2017JQ002）；安全绿色饲料添加剂中试中心项目（20160626002NY） |
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| 摘 要： | 试验旨在筛选长白山原始森林土壤中耐胁迫生长能力强、产酸能力强、体外抑菌特性好的耐低温乳酸菌，并对其相关生物学特性进行分析，为东北地区寒冬季节低温青贮提供优良乳酸菌菌种资源。采用低温（10℃）进行耐低温乳酸菌初筛，经不同pH、温度及盐度等条件下进行乳酸菌生长特性测定后，进行候选菌株的种属鉴定，并对候选菌株进行生长曲线、产酸曲线、耐酸和耐胆盐性能、抑菌性能测定及小鼠饲喂试验。结果显示，通过10℃低温条件培养，初筛获得11株能够良好生长的耐低温乳酸菌，其中C37、C34和C4212经16S rRNA测序分析后鉴定为植物乳杆菌，在培养7 d时，pH均可下降至4.25以下，均能够在4℃、45℃、pH 3.0、6.5% NaCl等条件下良好生长。生长曲线分析发现，3株乳酸菌均在4 h达到对数生长期，10~12 h后进入生长稳定期。在培养24 h时，C37、C34和C4212可将pH分别降至4.04以下，在pH 3.0条件下，C37和C4212具有较高的存活率，且对0.5%胆盐具有一定的耐受性，其存活率均在80%以上。对常见病原菌体外抑菌表明，3株乳酸菌对产肠毒素大肠杆菌、鼠伤寒沙门氏菌、金黄色葡萄球菌及肠炎沙门氏菌均有抑制作用，小鼠饲喂试验表明生物安全性好。本试验结果表明，从长白山原始森林土壤中筛选和鉴定出的乳杆菌C37、C34、C4212可作为东北地区寒冬季节低温青贮优良乳酸菌菌种资源。
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| 关 键 词： | 耐低温乳酸菌 筛选 植物乳杆菌 青贮 生物学特性 |
| 收稿时间： | 2020-04-21 |
Screening of Pychrotrophic Lactic Acid Bacteria from Changbai Mountain Soil |
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| CHEN Long,WEI Bingdong,ZHENG Lin,ZHANG Ying,YAN Xiaogang,YU Wei.Screening of Pychrotrophic Lactic Acid Bacteria from Changbai Mountain Soil[J].China Animal Husbandry & Veterinary Medicine,2020,47(10):3103-3113. |
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| Authors: | CHEN Long WEI Bingdong ZHENG Lin ZHANG Ying YAN Xiaogang YU Wei |
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| Institution: | Branch of Animal Husbandry, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, China |
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| Abstract: | The purpose of the experiment was to screen psychrotrophic lactic acid bacteria with strong stress-resistant growth ability,acid production ability and good bacteriostatic characteristics in vitro,and analyze its related biological characteristics,so as to provide excellent psychrotrophic lactic acid bacteria resources for low temperature silage in Northeast China.The culture condition of 10 ℃ was used to screen the psychrotrophic lactic acid bacteria.After the determination of the growth characteristics of Lactobacillus under different pH,temperature and salinity conditions,the species identification of the candidate strains was carried out,and the growth curve,acid production curve,acid and bile salt resistance,bacteriostatic performance and feeding test of mice were conducted for the candidate strains.The results showed that 11 strains of psychrotrophic lactic acid bacteria could grow well at 10 ℃.The strains C37,C34 and C4212 were identified as Lactobacillus plantarum by 16S rRNA sequencing analysis.The pH of C37,C34 and C4212 decreased to below 4.25 after 7 days of culture.In addition,C37,C34 and C4212 could grow well at 4 ℃,45 ℃,pH 3.0 and 6.5% NaCl.The analysis of growth curve showed that the three strains of Lactobacillus all had a fast growth rate,entered the logarithmic growth period at 4 h,and entered the stable growth period in 10-12 h after inoculation.In the aspect of acid production rate,C37,C34 and C4212 could reduce pH below 4.04 at 24 h of culture.At pH 3.0,C37 and C4212 had a high survival rate,they all had a certain tolerance to 0.5% bile salt,and the survival rate was more than 80%.C37,C34 and C4212 had inhibitory effect on enterotoxin producing Escherichia coli,Salmonella Typhimurium,Staphylococcus aureus and Salmonella Enteritis,and the animal test of three Lactobacillus plantarum were safe.Based on the results of this experiment,Lactobacillus C37,C34 and C4212 were screened and identified from the forest soil of Changbai Mountain,which could provide excellent lactic acid bacteria resources for low temperature silage in winter in Northeast China. |
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| Keywords: | psychrotrophic lactic acid bacteria screening Lactobacillus plantarum silage biological characteristics |
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