11株瘤胃源芽孢杆菌生理学特性研究

doi:10.16431/j.cnki.1671-7236.2018.04.008

摘要/Abstract

摘要：

试验对已筛选的11株瘤胃源芽孢杆菌的产酶活力、耐酸、耐胆盐和抑菌活性及菌株间相容性进行研究。采用3，5-二硝基水杨酸法（DNS法）测定纤维素酶的活力，涂布平板计数法测定菌株的耐酸性、耐胆盐等特性，牛津杯法测定抑菌活性，滤纸片法进行室内相容性试验。结果显示，菌株T-2、T-3和T-10产纤维素酶的酶活力较高；菌株T-2、T-5、T-7、T-8和T-9表现较强的耐酸能力；菌株T-4、T-5、T-9和T-11表现较强的耐胆盐能力。菌株T-4和T-7对副溶血弧菌的抑菌活性较强，T-7对产气杆菌的抑菌活性最强，T-5、T-9、T-10对大肠杆菌抑菌活性较强，而T-3对藤黄八叠球菌指示菌的抑菌性最强。室内相容性试验结果表明，11株芽孢杆菌部分菌株之间存在颉颃作用，由此筛选出优良菌株的复配组合，抑菌可能的最优复配组合为T-3、T-4、T-5；T-3、T-4、T-10；T-4、T-5、T-10；T-3、T-7、T-10。纤维素酶活力较强时可能的最优复配组合为T-2、T-3、T-5；T-2、T-3、T-10；T-2、T-5、T-10；T-3、T-5、T-6；T-3、T-5、T-10。本试验可为动物饲料的研究和开发提供一定的参考依据。

关键词: 芽孢杆菌; 生物学特性; 耐受性; 相容性

Abstract:

The biological characteristics of eleven Bacillus strains from rumen were studied,including the activity of produced enzyme,the endurance to acid and bile salt,the activity of antibacteria and the compatibility.3,5-dinitrosalicylic acid colorimetric method (DNS method) was used to determine the enzyme activity,acid resisting test and bile salt resisting test were conducted by plate spreading method,antibacterial activity was determiend by Oxford cup method,and while indoor compatibility experiment was studied by filter paper method.The results showed that the activity of cellulase produced by T-2,T-3 and T-10 were higher;T-2,T-5,T-7,T-8 and T-9 had stronger acid resistance;T-4,T-5,T-9 and T-11 had stronger ability of bile salt-tolerance;T-4 and T-7 had stronger antibacterial activity against Vibrio parahaemolyticus,and T-7 had strongest antibacterial activity against Aerogenes,T-5,T-9 and T-10 had stronger antibacterial activity against Escherichia coli and T-3 had strongest antibacterial activity against Sarcina lutea.Indoor compatibility experiments showed that there were antagonistic effects among some strains of eleven Bacillus strains.The combinations of the selected strains for bacteriostat were T-3,T-4,T-5;T-3,T-4,T-10;T-4,T-5,T-10;T-3,T-7,T-10.The combinations of the selected strains for higher cellulase activities were T-2,T-3,T-5;T-2,T-3,T-10;T-2,T-5,T-10;T-3,T-5,T-6;T-3,T-5,T-10.The results could provide some references for the research and development of animal feed.

Key words: Bacillus; biological characteristics; tolerance; compatibility

中图分类号:

Q936

谈诗谣, 张雪, 王红英, 钱斯日古楞. 11株瘤胃源芽孢杆菌生理学特性研究[J]. 《中国畜牧兽医》, 2018, 45(4): 898-904.

TAN Shiyao, ZHANG Xue, WANG Hongying, QIANSIRI Guleng. Biological Characteristics of Eleven Bacillus Strains from Rumen[J]. , 2018, 45(4): 898-904.

参考文献

[1] MARTINEZ J L.Environmental pollution by antibiotics and by antibiotic resistance determinants[J].Environmental Pollution,2009,157(11):2893-2902.
[2] 高立红,史亚利,厉文辉,等.抗生素环境行为及其环境效应研究进展[J].环境化学,2013,32(9):1619-1628. GAO L H,SHI Y L,LI W H,et al.Environmental behavior and impacts of antibiotics[J].Environmental Chemistry,2013,32(9):1619-1628.(in Chinese)
[3] 刘开朗,王加启,卜登攀.2008-2009年反刍动物营养研究进展Ⅰ.瘤胃微生物多样性与功能[J].中国畜牧兽医,2010,37(2):5-14. LIU K L,WANG J Q,BU D P.Annual review of ruminant nutrition in 2008-2009Ⅰ.Diversity and function of rumen microflora[J].China Animal Husbandry & Veterinary Medicine,2010,37(2):5-14.(in Chinese)
[4] 邓丽,芮汉明.益生菌的研究进展[J].现代食品科技,2003,19:84-86. DENG L,RUI H M.Research progress of probio-tics[J].Modern Food Science and Technology,2003,19:84-86.(in Chinese)
[5] SURAJIT S.Probiotics as functional foods:Gut colonization and safety concerns[J].Nutrition & Food Science,2013,43(5):496-504.
[6] 孔健.农业微生物技术[M].北京:化学工业出版社,2005. KONG J.Agriculture Microbial Technology[M].Beijing:Chemical Industry Press,2005.(in Chinese)
[7] 肖宏德,肖运才,何熙贞,等.用益生菌制剂代替断奶仔猪日粮中抗生素的研究[J].动物医学进展,2014,35(3):53-58. XIAO H D,XIAO Y C,HE X Z,et al.Study on probiotic agents substituting for antibiotics in weaned piglet diets[J].Progress in Veterinary Medicine,2014,35(3):53-58.(in Chinese)
[8] SAMANYA M,YAMAUCHI K E.Histological alterations of chickens fed dried Bacillus subtilis var natto[J].Camparative Biochemistry and Physiology Part A,Molecular & Integrative Physiology,2002,133:496-504.
[9] 张新雄,彭锋,毛光平,等.饲用芽孢杆菌研究与应用进展[J].应用与环境生物学报,2013,19(5):891-897. ZHANG X X,PENG F,MAO G P,et al.Advance in research and application of Bacillus probiotics[J].Chinese Journal of Applied and Environmental Biology,2013,19(5):891-897.(in Chinese)
[10] 袁道强,黄建华.生物化学实验和技术[M].北京:中国轻工业出版社,2006. YUAN D Q,HUANG J H.Biochemical Experiments and Techniques[M].Beijing:China Light Industry Press,2006.(in Chinese)
[11] 张全国,张志萍,赵民善,等.秸秆制氢过程中纤维素酶酶活测定方法研究[J].热科学与技术,2011,10(1):128-132. ZHANG Q G,ZHANG Z P,ZHAO M S,et al.Studies on new DNS method for determination of filter paper enzyme activity in straw hydrogen production process[J].Journal of Thermal Science and Technology, 2011,10(1):128-132.(in Chinese)
[12] 崔海洋,程仕伟,黄田红,等.产纤维素酶的解淀粉芽孢杆菌分离鉴定及酶学性质研究[J].食品科学技术学报,2014,32(3):43-47. CUI H Y,CHENG S W,HUANG T H,et al.Screening and identifying of Bacillus amyloliquefaciens for cellulase production and its enzymatic characters[J].Journal of Food Science and Technology,2014, 32(3):43-47.(in Chinese)
[13] DASHTBAN M,BUCHKOWSKI R,QIN W.Effect of different carbon sources on cellulase production by Hypocrea jecorina (Trichoderma reesei) strains[J].International Journal Biochemistry & Molecular Biology,2011,2(3):274-286.
[14] 宁豫昌,赵绪永,郑鸣.猪源芽孢杆菌的分离鉴定及生物学特性研究[J].中国畜牧兽医,2012,39(9):65-68. NING Y C,ZHAO X Y,ZHENG M.Isolation,identification of Bacillaceae from swine and study on its biological characteristics[J]. China Animal Husbandry & Veterinary Medicine,2012,9(9):65-68.(in Chinese)
[15] 李善仁,陈济琛,蔡海松,等.三株芽孢杆菌作为益生菌的生物特性[J].营养学报,2010,32(1):75-78. LI S R,CHEN J C,CAI H S, et al.The properties of three Bacillus strains as potential probiotics[J].Acta Nutrimenta Sinica,2010,32(1):75-78.(in Chinese)
[16] 钟罗华,邓舜洲,张文波,等.6株猪源芽孢杆菌的分离鉴定及其益生性研究[J].中国畜牧兽医,2017,44(2):521-529. ZHONG L H,DENG S Z,ZHANG W B,et al.Isolation and identification of six strains of swine Bacillus and their probiotic research[J]. China Animal Husbandry & Veterinary Medicine,2017,44(2):521-529.(in Chinese)
[17] 佘之蕴,黄宝莹,刘海卿,等.牛津杯法测定食品添加剂对五种益生菌的抑菌活力[J].食品工业,2016,37(1):171-173. SHE Z Y,HANG B Y,LIU H Q,et al.Antibacterial activity of foods additives against five strains of probiotics by oxford plate assay system[J].The Food Industry,2016,37(1):171-173.(in Chinese)
[18] 刘秀侠,徐海燕,辛国芹,等.11株枯草芽孢杆菌益生特性研究[J].中国畜牧兽医,2017,8(44):2333-2341. LIU X X,XU H Y,XIN G Q,et al.Study on probiotic characteristics of eleven Bacillus subtilis strains[J]. China Animal Husbandry & Veterinary Medicine,2017,8(44):2333-2341.(in Chinese)
[19] 陈亮,黄亮,李静琴.不同芽孢杆菌对雏鸡细菌性腹泻病原菌的体外抑菌试验[J].饲料工业,2011,32(18):56-57. CHEN L,HUANG L,LI J Q.In vitro antibacterial test of different Bacillus strains on pathogenic bacteria of bacterial diarrhea in chickens[J].Feed Industry,2011,32(18):56-57.(in Chinese)
[20] 张艳群,来航线,韦小敏,等.生物肥料多功能芽孢杆菌的筛选及其作用机理研究[J].植物营养与肥料学报,2013,19(2):489-497. ZHANG Y Q,LAI H X,WEI X M, et al.Selection of multifunctional Bacillus strains of biological fertilizer and the study of their mechanisms on plant growth[J].Journal of Plant Nutrition and Fertilizer,2013,19(2):489-497.(in Chinese)
[21] 张丽青,吴海龙,姜红霞,等.纤维素降解细菌的筛选及其产酶条件优化[J].环境科学与管理,2007,32(10):110-117. ZHANG L Q,WU H L,JIANG H X, et al.Screening on bacteria capable of degrading cellulose and optimizing its conditions for cellulase production[J]. Environmental Science and Management,2007,32(10):110-117.(in Chinese)
[22] 姚勇芳,廖延智,陈小凤,等.枯草芽孢杆菌Y-3生长特性及抗性研究[J].安徽农业科学,2013,41(2):11395-11398. YAO Y F,LIAO Y Z,CHEN X F,et al.Studies on the growth characteristies and resistance of Bacillus subtilis Y-3[J].Journal of Anhui Agricultural Sciences,2013,41(2):11395-11398.(in Chinese)
[23] 谭才邓,朱美娟,杜淑霞,等.抑菌试验中抑菌圈法的比较研究[J].食品工业,2016,11(37):122-125. TAN C D,ZHU M J,DU S X,et al.Study on the inhibition zone method in antimicrobial test[J].The Food Industry,2016,11(37):122-125.(in Chinese)
[24] 王春芳,马诗淳,黄艳,等.降解水稻秸秆的复合菌系及其微生物群落结构演替[J].微生物学报,2016,56(12):1856-1868. WANG C F,MA S C,HANG Y,et al.Characterization and microbial community shifts of rice strawdegrading microbial consortia[J].Acta Microbiologica Sinica,2016,56(12):1856-1868.(in Chinese)