研究论文 RESEARCH PAPER

青贮用乳酸菌的分离鉴定及生物学特性评价

  • 韦庆旭 ,
  • 张建鹏 ,
  • 梁煜晨 ,
  • 唐连群 ,
  • 王平
展开
  • 1. 西北农林科技大学动物科技学院, 杨凌 712100;
    2. 新疆农业大学动物科技学院, 乌鲁木齐 830002
韦庆旭(1996-),女,吉林桦甸人,硕士研究生,畜牧专业。E-mail:weiqingxu0125@163.com

收稿日期: 2021-12-21

  网络出版日期: 2022-07-14

基金资助

陕西省重点研发计划(2018ZDXM-NY-033);陕西省科技创新驱动项目(NYKJ-2019-YL24)

Isolation, Identification and Biological Characteristics Evaluation of Lactic Acid Bacteria for Silage

  • WEI Qingxu ,
  • ZHANG Jianpeng ,
  • LIANG Yuchen ,
  • TANG Lianqun ,
  • WANG Ping
Expand
  • 1. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China;
    2. College of Animal Science and Technology, Xinjiang Agricultural University, Urumqi 830002, China

Received date: 2021-12-21

  Online published: 2022-07-14

摘要

本研究旨在从构树青贮、玉米青贮及饲喂构树青贮的羔羊粪便中分离鉴定可饲用乳酸菌,为开发构树青贮用乳酸菌提供试验材料。以构树青贮、玉米青贮以及饲喂构树青贮的羔羊粪便为菌株分离来源,使用含有1%碳酸钙的MRS琼脂培养基分离纯化乳酸菌,获得菌株经16S rRNA测序鉴定后对其中可饲用菌株进行生长、产酸性能的筛选,评估在pH 3.0、3.5、4.0条件下存活率和对大肠杆菌、金黄色葡萄球菌和沙门氏菌的抑制能力,并对筛选菌株进行菌株之间拮抗性测定。结果表明:获得可饲用乳酸菌共37株,包括植物乳杆菌(Lactobacillus plantarum)、布氏乳杆菌(Lactobacillus buchneri)、副干酪乳杆菌(Lactobacillus paracasei)、粪肠球菌(Enterococcus faecalis)4种,不同的菌株在生长和产酸性能上表现出差异,在不同pH中存活率和抑菌性能也具有菌株特异性。综合比较不同菌株之间的生物学特性,植物乳杆菌GQ-3-2、GQ-3-29及GQ-5-4具有良好的生长、产酸及抑菌性能;粪肠球菌Y-3-15在发酵2 h进入对数生长期,能够为植物乳杆菌的大量繁殖创造良好环境;布氏乳杆菌GQ-3-21产酸性能良好,在发酵0~16 h pH快速下降,在pH 4.0的条件下存活率大于50%,在pH 3.5的条件下也能够微弱存活,是有潜力的青贮接种菌。最终选择菌株GQ-3-2、GQ-3-29、GQ-5-4、Y-3-15和GQ-3-21作为青贮用乳酸菌复合制剂的候选菌株,5株候选菌株之间不会互相抑制生长。综上所述,本试验筛选获得5株可饲用乳酸菌,其生物学特性良好,菌株之间无拮抗作用,可作为后续开发构树青贮饲料的乳酸菌复合制剂菌种来源。

本文引用格式

韦庆旭 , 张建鹏 , 梁煜晨 , 唐连群 , 王平 . 青贮用乳酸菌的分离鉴定及生物学特性评价[J]. 动物营养学报, 2022 , 34(7) : 4737 -4749 . DOI: 10.3969/j.issn.1006-267x.2022.07.060

Abstract

The objective of this study was to isolate and identify the edible lactic acid bacteria from the feces of Broussonetia papyrifera silage, corn silage and feces of kids fed with Broussonetia papyrifera silage, and to provide experimental materials for developing lactic acid bacteria for Broussonetia papyrifera silage. Using Broussonetia papyrifera silage, corn silage and feces of kids fed with Broussonetia papyrifera silage as the sources of strain isolation, the lactic acid bacteria were isolated and purified by MRS agar medium containing 1% calcium carbonate. The obtained strains were identified by 16S rRNA sequencing, and then the feed strains were screened for growth and acid producing performance, the survival rate and inhibition ability to Escherichia coli, Staphylococcus aureus and Salmonella under the conditions of pH 3.0, 3.5 and 4.0 were evaluated, and the antagonistic ability among screened strains was determined. The results showed that 37 strains of edible lactic acid bacteria were obtained, including Lactobacillus plantarum, Lactobacillus buchneri, Lactobacillus paracasei and Enterococcus faecalis. Different strains showed differences in growth and acid-producing performance, and showed strain specificity in survival rate and antibacterial performance in different pH. A comprehensive comparison of the biological characteristics of different strains showed that Lactobacillus plantarum GQ-3-2, GQ-3-29 and GQ-5-4 had good growth, acid production and antibacterial performance; Enterococcus faecalis Y-3-15 entered the logarithmic growth phase after 2 hours of fermentation, which could create a good environment for the mass reproduction of Lactobacillus plantarum; Lactobacillus brucellosis GQ-3-21 had good acid production, and pH could decrease rapidly in 0 to 16 hours of fermentation, the survival rate was more than 50% under the condition of pH 4.0, and it could survive weakly under the condition of pH 3.5, so it is a potential silage inoculation bacteria. Finally, strains GQ-3-2, GQ-3-29, GQ-5-4, Y-3-15 and GQ-3-21 were selected as candidate strains of lactic acid bacteria compound preparation for silage, and the five candidate strains could not inhibit each other's growth. In conclusion, five strains of edible lactic acid bacteria are screened in this experiment, which have good biological characteristics, and there is no antagonism among them, it can be used as the source of lactic acid bacteria compound bacteria for the subsequent development of Broussonetia papyrifera silage.

参考文献

[1] 张志清.乳酸乳球菌异源表达漆酶及在玉米秸秆青贮应用[D].硕士学位论文.呼和浩特:内蒙古大学,2020. ZHANG Z Q.Construction of recombinant Lactococcus lactis with laccase secreting ability and its application in silage of corn stalk[D].Master's Thesis.Hohhot:Inner Mongolia University,2020.(in Chinese)
[2] ALI N,WANG S R,ZHAO J,et al.Microbial diversity and fermentation profile of red clover silage inoculated with reconstituted indigenous and exogenous epiphytic microbiota[J].Bioresource Technology,2020,314:123606.
[3] 司丙文,徐文财,张小利,等.不同添加剂对杂交构树青贮发酵品质的影响[J].动物营养学报,2018,30(11):4670-4675. SI B W,XU W C,ZHANG X L,et al.Influence of different additives on fermentation quality of hybrid paper mulberry silage[J].Chinese Journal of Animal Nutrition,2018,30(11):4670-4675.(in Chinese)
[4] 王德光.饲用菌种(株)的分离鉴定及复合菌剂降解玉米皮-统糠粗纤维的效果研究[D].硕士学位论文.杨凌:西北农林科技大学,2017. WANG D G.Isolation and identification of feeding strains and the effect of compound bacteria on degradation of corn bran chitosan[D].Master's Thesis.Yangling:Northwest A&F University,2017.(in Chinese)
[5] 韩吉雨.青贮发酵体系中乳酸菌多样性的研究[D].博士学位论文.呼和浩特:内蒙古农业大学,2009. HAN J Y.Study on diversity of lactic acid bacteria in silage fermentation[D].Ph.D.Thesis.Hohhot:Inner Mongolia University,2009.(in Chinese)
[6] TIMMERMAN H M,VELDMAN A,VAN DEN ELSEN E,et al.Mortality and growth performance of broilers given drinking water supplemented with chicken-specific probiotics[J].Poultry Science,2006,85(8):1383-1388.  
[7] 鲍宇红.西藏青贮用乳酸菌筛选分离及应用研究进展[J].畜牧与饲料科学,2020,41(6):37-42. BAO Y H.Research advances on the screening,isolation and application of lactic acid bacteria for silage in Tibet[J].Animal Husbandry and Feed Science,2020,41(6):37-42.(in Chinese)
[8] 王隆,李璟怡,欧阳可寒,等.不同青贮添加剂对去油芳樟枝叶青贮饲料营养成分、青贮发酵品质和瘤胃体外发酵特性的影响[J/OL].动物营养学报:1-11.(2021-12-24)[2022-01-31].http://kns.cnki.net/kcms/detail/11.5461.S.20211223.1125.002.html. WANG L,LI J Y,OUYANG K H,et al.Effects of different silage additives on nutrient composition,silage fermentation quality and rumen fermentation characteristics in vitro of deoiling linalyl branches and leaves silage[J/OL].Chinese Journal of Animal Nutrition:1-11.(2021-12-24)[2022-01-31].http://kns.cnki.net/kcms/detail/11.5461.S.20211223.1125.002.html. (in Chinese)
[9] 朱江江.奶山羊脂肪酸合酶基因乙酰/丙二酸单酰转移酶区域的过表达研究[D].硕士学位论文.杨凌:西北农林科技大学,2011. ZHU J J.Overexpression of mat domain of fatty acid synthase (FASN) gene in dairy goat[D].Master's Thesis.Yangling:Northwest A&F University,2011.(in Chinese)
[10] 韦票.瘤胃源饲用微生物的筛选及复合菌剂发酵中药渣效果的研究[D].硕士学位论文.杨凌:西北农林科技大学,2021. WEI P.Study on the screening of ruminal direct-fed microorganisms and the effect of compound bacteria on the fermentation of Chinese medicine residue[D].Master's Thesis.Yangling:Northwest A&F University,2021.(in Chinese)
[11] 何江波,姚志芳,吴国芳,等.羊瘤胃源乳酸菌的分离鉴定及其生物学特性分析[J].动物营养学报,2021,33(6):3365-3379. HE J B,YAO Z F,WU G F,et al.Isolation and identification of lactic acid bacteria from sheep rumen and analysis of their biological characteristics[J].Chinese Journal of Animal Nutrition,2021,33(6):3365-3379.(in Chinese)
[12] 高擎燏,李平华,黄瑞华,等.猪源乳酸菌的抗逆性及益生性研究[J].畜牧与兽医,2015,47(10):41-46. GAO Q Y,LI P H,HUANG R H,et al.Study on stress resistance and probiotics of lactic acid bacteria from pigs[J].Animal Husbandry & Veterinary Medicine,2015,47(10):41-46.(in Chinese)
[13] 王蔚淼,潘玲,朱玮,等.10株乳酸菌产酸性能的研究[J].中国生态农业学报,2007,15(3):207-208. WANG W M,PAN L,ZHU W,et al.Acid-producing characteristics of ten lactic acid bacteria[J].Chinese Journal of Eco-Agriculture,2007,15(3):207-208.(in Chinese)
[14] 靳胜男,舒慧萍,张冬星,等.2株猪源乳酸杆菌的抗逆性和生长特性分析[J].吉林农业大学学报,2021,43(6):734-740. JIN S N,SHU H P,ZHANG D X,et al.Stress resistance and growth characteristics of two strains from swine Lactobacillus[J].Journal of Jilin Agricultural University,2021,43(6):734-740.(in Chinese)
[15] 张红梅,段珍,李霞,等.青贮饲料乳酸菌添加剂的应用现状[J].草业科学,2017,34(12):2575-2583. ZHANG H M,DUAN Z,LI X,et al.Actual research and application of the silage lactic acid bacteria additives[J].Pratacultural Science,2017,34(12):2575-2583.(in Chinese)
[16] 蔡义民,熊井清雄,廖芷,等.乳酸菌剂对青贮饲料发酵品质的改善效果[J].中国农业科学,1995(2):73-82. CAI Y M,XIONGJING Q X,LIAO Z,et al.Effect of lactic acid bacteria inoculants on fermentative quality of silage[J].Scientia Agricultura Sinica,1995(2):73-82.(in Chinese)
[17] 白长胜,孟维珊,李进,等.自然界中大豆秸秆发酵用优良乳酸菌的分离筛选[J].现代畜牧科技,2020(10):8-11. BAI C S,MENG W S,LI J,et al.Isolation and screening of excellent lactic acid bacteria for soybean straw fermentation from nature[J].Technical Advisor for Animal Husbandry,2020(10):8-11.(in Chinese)
[18] 雷晓青,孟阳,王智敏,等.发酵饲料中抑菌菌种筛选及抑菌性能评估方法的初步研究[J].饲料研究,2021,44(3):65-68. LEI X Q,MENG Y,WANG Z M,et al.Screening of antibacterial strains in fermented feed and preliminary study on evaluation method of antibacterial performance[J].Feed Research,2021,44(3):65-68.(in Chinese)
[19] 郑昕,唐霄,宫利宏,等.鸡源肠道乳酸菌的筛选及抑菌作用分析[J].养禽与禽病防治,2020(12):9-15. ZHENG X,TANG X,GONG L H,et al.Screening of Lactobacillus from chicken intestine and analysis of antibacterial effect[J].Poultry Husbandry and Disease Control,2020(12):9-15.(in Chinese)
[20] LEROY F,DE VUYST L.Lactic acid bacteria as functional starter cultures for the food fermentation industry[J].Trends in Food Science & Technology,2004,15(2):67-78.  
[21] SI H Z,LIU H L,LI Z P,et al.Effect of Lactobacillus plantarum and Lactobacillus buchneri addition on fermentation,bacterial community and aerobic stability in lucerne silage[J].Animal Production Science,2018,59(8):1528-1536.
[22] TODOROV S D,DICKS L M T.Bacteriocin production by Lactobacillus pentosus ST712BZ isolated from boza[J].Brazilian Journal of Microbiology,2007,38(1):166-172.  
文章导航

/