广西猪源益生菌的分离鉴定与生物学特性研究

doi:10.16431/j.cnki.1671-7236.2020.06.032

Abstract

Abstract: In order to investigate the prevalence of probiotics in pig feces and microecological fermented feeds in Guangxi area,and to rationally develop and utilize porcine probiotics,in this experiment,15 pig feces samples and 5 probiotic fermentation feeds were collected and the probiotic bacteria were isolated.The morphological characteristics,physiological and biochemical characteristics and 16S rRNA molecular sequence of the probiotics were studied.The growth curve,bile salt resistance test,temperature sensitivity test,gastrointestinal tolerance test,in vitro and in vivo bacteriostasis test,drug resistance test,mouse safety test were carried out to study the biological characteristics of the isolated strains.In this experiment,6 species and 29 probiotics strains were isolated and identified,including 1 Enterococcus faecium (C2),2 Enterococcus lactate (C1 and C3),3 Lactobacillus plantarum (R20,R30 and R37),4 Lactobacillus casei (R41-R44),7 Bacillus subtilis (K1-K7) and 12 Bacillus cereus (Y1-Y12).Six probiotics with good growth performance,bile salt resistance,high temperature resistance,gastrointestinal tolerance and strong antibacterial ability in vitro were screened out,including Enterococcus lactis C1,Enterococcus faecium C2,Lactobacillus plantarum R20,Lactobacillus casei R41,Bacillus subtilis K1 and Bacillus cereus Y3.The safety test of the selected strains in mice showed that six probiotics were safe and harmless to mice at the concentration of 10⁹ CFU/mL.At the same time,C2,R20,R41 and Y3 probiotics could extremely significantly increase the weight of mice (P<0.01),K1 could markedly increase the weight of mice (P<0.05).The results of bacteriostatic experiment in vivo showed that C2,R20 and K1 could reduce the mortality of mice infected by Salmonella G21,and had a certain degree of antibacterial effect in vivo.All test results showed that R20 and K1 could be selected as pig probiotic strains.

Key words: probiotics; isolation; biological characteristics; bacteriostasis; swine

CLC Number:

S816.3

ZHANG Zai, ZHENG Ruicheng, FANG Qingli, HUANG Weijian. Isolation,Identification and Biological Characteristics of Probiotics from Swine Origin in Guangxi[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1921-1933.

References

[1] 高阳,王海岩,王佳江,等.益生菌的保健作用与研究综述[J].安徽农学通报,2009,15(19):56. GAO Y,WANG H Y,WANG J J,et al.Health function and research direction of probiotics[J].Anhui Agricultural Science Bulletin,2009,15(19):56.(in Chinese)
[2] 刘健华,陈杖榴.动物性食品中抗菌药残留对人体肠道菌群的影响及其研究模型与方法[J].动物医学进展,2003,24(2):37-40. LIU J H,CHEN Z L.Effects of antimicrobial drug residues from food of animal origin on the human intestinal flora[J].Progress in Veterinary Medicine,2003,24(2):37-40.(in Chinese)
[3] 高霞,李卫,涂世.益生菌的功效及益生菌乳制品[J].广东化工,2009,36(5):122-123. GAO X,LI W,TU S.The Physiological,effects of probiotics and their dairy products[J].Guangdong Chemical Industry,2009,36(5):122-123.(in Chinese)
[4] MANDAL S,HATI S.Microencapsulation of bacterial cells by emulsion technique for probiotic application[J].Methods in Molecular Biology,2017,1479:273-279.
[5] BARATHIKANNAN K,CHELLIAH R,RUBAB M,et al.Gut microbiome modulation based on probiotic application for anti-obesity:A review on efficacy and validation[J].Microorganisms,2019,7(10):E456.
[6] AHMEDBEYLI D R,SEYIDBEKOV O S,DIRIKAN I S,et al.Efficacy of probiotic application in the treatment and prevention of peri-implant mucositis[J].Stomatologiia (Mosk),2019,98(4):20-24.
[7] 时云朵,任燕,孙豪.益生菌潜在安全性的现状及对策[J].饲料博览.2018,5:43-46. SHI Y D,REN Y,SUN H.Probiotics potential safety and countermeasures[J].Feed Review,2018,5:43-46.(in Chinese)
[8] 郭志杰,李杰,李鑫,等.猪源乳酸菌的分离、筛选、鉴定及产酸性能的研究[J].饲料工业,2010,31(20):45-48. GUO Z J,LI J,LI X,et al.Isolation and identification of lactic acid bacteria and study on their capability of lactic acid production[J].Feed Industry,2010,31(20):45-48.(in Chinese)
[9] 赵振宇.猪源蜡样芽孢杆菌的分离鉴定[D].长沙:湖南农业大学,2014. ZHAO Z Y.Isolation and characterization of Bacillus cereus from piglet[D].Changsha:Hunan Agricultural University,2014.(in Chinese)
[10] 蔡妙英,东秀珠.常见细菌系统鉴定手册[M].北京:科学出版社,2001. CAI M Y,DONG X Z.Handbook of Systematic Identification of Common Bacteria[M].Beijing:Science Press,2001.(in Chinese)
[11] BERGEY D H,HOLT J G,KRIEG N R.Bergey's Manual of Systematic Bacteriology[M].Springer:Bergey's Manual of Systematic Bacteriology,1984.
[12] CONWAY P L,GORBACH S L,GOLDIN B R.Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells[J].Journal of Dairy Science,1987,70(1):1-12.
[13] 李治华,黄驰,王自鹏,等.发酵豆瓣酱微生物宏基因组DNA提取研究[J].西南农业学报,2013,26(6):2663-2665. LI Z H,HUANG C,WANG Z P,et al.Study on extraction of metagenomic DNA related to broad-bean sauce fermentation[J].Southwest China Journal of Agricultural Sciences,2013,26(6):2663-2665.(in Chinese)
[14] 凌代文,东秀珠.乳酸细菌分类鉴定及实验方法[J].微生物学通报,1999.26(1):23. LING D W,DONG X Z.Classification,identification and experimental methods of lactic acid bacteria[J].Microbiology China,1999,26(1):23.(in Chinese)
[15] 孙长贵,成军,杨燕.2012年CLSI M100-S22文件主要更新内容介绍[J].浙江检验医学,2012,10(2):58. SUN C G,CHENG J,YANG Y.Introduction to main updated contents of CLSI M100-S22 document in 2012[J].Zhejiang Journal of Laboratory Medicine,2012,10(2):58.(in Chinese)
[16] 杨维红.Ⅰ型菌毛对鼠伤寒沙门氏菌体内侵袭能力的影响[D].武汉:华中农业大学,2007. YANG W H.Effect of type Ⅰ pili on the invasiveness of Salmonella typhimurium in vivo[D].Wuhan:Huazhong Agricultural University,2007.(in Chinese)
[17] 刘昌孝.当代抗生素发展的挑战与思考[J].中国抗生素杂志,2017,42(1):1-12. LIU C X.Challenges and thinking of current antibiotic development[J].Chinese Journal of Antibiotics,2017,42(1):1-12.(in Chinese)
[18] 马晨,张和平.益生菌、肠道菌群与人体健康[J].科技导报,2017,35(21):14-25. MA C,ZHANG H P.Probiotic,intestinal microbiota and human health[J].Science & Technology Review,2017,35(21):14-25.(in Chinese)
[19] 张秀林,魏小兵,欧长波,等.益生菌发酵饲料对仔猪生长和免疫功能影响的研究进展[J].中国畜牧兽医,2017,44(2):476-481. ZHANG X L,WEI X B,OU C B,et al.Research Progress of Probiotics fermented feed effects on growth performance and immune function in piglets[J].China Animal Husbandry & Veterinary Medicine,2017,44(2):476-481.(in Chinese)
[20] 翟云,托娅.益生菌的免疫调节作用及其相关应用研究进展[J].中国微生态学杂志,2018,30(2):235-239. ZHAI Y,TUO Y.The immunomodulatory effects of probiotics and recent advances in related researches[J].Chinese Journal of Microecology,2018,30(2):235-239.(in Chinese)
[21] 陈忠秀,李嘉文,赵扬,等.益生菌的应用现状和发展前景[J].中国微生态学杂志,2016,28(4):493-497. CHEN Z X,LI J W,ZHAO Y,et al.Probiotics:Current application status and development prospect[J].Chinese Journal of Microecology,2016,28(4):493-497.(in Chinese)
[22] 祝天龙,李奎,邵强,等.枯草芽孢杆菌制剂对弱仔猪生长性能、免疫指标的影响[J].黑龙江畜牧兽医,2015,11:201-204. ZHU T L,LI K,SHAO Q,et al.Effect of Bacillus subtilis preparation on growth performance and immune index of weak piglets[J].Heilongjiang Animal Science and Veterinary Medicine,2015,11:201-204.(in Chinese)
[23] YANG C M,FERKET P R,HONG Q H,et al.Effect of chito-oligosaccharide on growth performance,intestinal barrier function,intestinal morphology and cecal microflora in weaned pigs[J].Journal of Animal Science,2012,90(8):2671-2676.
[24] 祝永才,胡兴义,张双翔,等.复合益生菌对早期断奶仔猪生长性能和免疫水平的影响[J].中国畜牧兽医,2018,45(6):1518-1525. ZHU Y C,HU X Y,ZHANG S X,et al.Effects of compound probiotics on growth performance and immune levels of early weaned pigs[J].China Animal Husbandry & Veterinary Medicine,2018,45(6):1518-1525.(in Chinese)
[25] 叶红玲,刘冬,叶明,等.复合益生菌对昆明小鼠生长性能和肠道酶活性的影响[J].安徽农业科学,2016,44(19):173-175. YE H L,LIU D,YE M,et al.Effects of compound probiotics on growth performance and intestinal protease activity in Kunmin mice[J].Journal of Anhui Agricultural Sciences,2016,44(19):173-175.(in Chinese)
[26] 曾佳佳,陈静,陈振,等.中草药-益生菌-胆汁酸复合制剂对蛋鸡生产性能、蛋品质及免疫指标的影响[J].中国畜牧兽医,2018,45(4):940-946. ZENG J J,CHEN J,CHEN Z,et al.Effect of Chinese herbal-probiotics-bile acids compound on production performance,egg quality and immune indexes of laying hens[J].China Animal Husbandry & Veterinary Medicine,2018,45(4):940-946.(in Chinese)
[27] 杨坤明,任航,苏雅婷,等.几种商用益生芽孢杆菌特性的比较研究[J].中国畜牧兽医,2018,45(5):1241-1250. YANG K M,REN H,SU Y T,et al.Comparative evaluation of probiotic characterization of commercial Bacillus products[J].China Animal Husbandry & Veterinary Medicine,2018,45(5):1241-1250.(in Chinese)

Isolation,Identification and Biological Characteristics of Probiotics from Swine Origin in Guangxi

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