研究论文 RESEARCH PAPER

鸡源卷曲乳杆菌Y68的分离鉴定及其体外益生特性评估

  • 张家宝 ,
  • 马轲 ,
  • 刘真真 ,
  • 高宇 ,
  • 蔺小奇 ,
  • 杨勇军
展开
  • 吉林大学动物医学学院, 长春 130062
张家宝(1998—),女,吉林蛟河人,硕士研究生,研究方向为预防兽医学。E-mail:18943839416@163.com

收稿日期: 2021-09-18

  网络出版日期: 2022-05-14

基金资助

国家自然科学基金项目(31972682)

Isolation, Identification of Lactobacillus crispatus Y68 from Chickens and Evaluation of Its in Vitro Probiotics Properties

  • ZHANG Jiabao ,
  • MA Ke ,
  • LIU Zhenzhen ,
  • GAO Yu ,
  • LIN Xiaoqi ,
  • YANG Yongjun
Expand
  • College of Veterinary Medicine, Jilin University, Changchun 130062, China

Received date: 2021-09-18

  Online published: 2022-05-14

摘要

本试验旨在筛选研制畜禽微生态制剂——乳酸菌,探究卷曲乳杆菌Y68的益生特性。通过抑菌能力筛查、生长曲线测定、产酸性能、药敏试验、体外模拟胃肠道条件耐受试验、自凝集试验以及疏水性试验,评估了其益生特性。结果表明:卷曲乳杆菌Y68对鸡白痢沙门氏菌ATCC19945、金黄色葡萄球菌USA300均有较强的抑制作用,抑菌圈直径分别为(21.64±0.31) mm、(15.13±0.23) mm;卷曲乳杆菌Y68对数生长期为6~24 h;经培养48 h后菌液pH稳定在3.89;卷曲乳杆菌Y68对红霉素、青霉素、头孢菌素、氯霉素、万古霉素、克林霉素较为敏感,对诺氟沙星、氧氟沙星、四环素、庆大霉素、呋喃唑酮表现出耐药;卷曲乳杆菌Y68在pH 2.0环境下孵育3 h存活率为52.61%,在人工胃液环境下3 h存活率为65.92%,在人工肠液环境下10 h时存活率仍为86.23%,能够耐受0.3%的胆盐环境;卷曲乳杆菌Y68自聚集率在2、24 h时分别为(29.71±0.40)%、(97.23±0.76)%,疏水率为(89.59±0.63)%。综上所述,卷曲乳杆菌Y68具有较强的产酸能力、抑菌能力,对模拟的胃肠道条件具有耐受性,包括低pH、胃蛋白酶、胰酶和胆盐环境,且保持了较强的自聚集能力、疏水性能。鸡源卷曲乳杆菌Y68可作为畜禽微生态制剂的候选菌株,应用于畜禽营养与饲料中。

本文引用格式

张家宝 , 马轲 , 刘真真 , 高宇 , 蔺小奇 , 杨勇军 . 鸡源卷曲乳杆菌Y68的分离鉴定及其体外益生特性评估[J]. 动物营养学报, 2022 , 34(5) : 3339 -3347 . DOI: 10.3969/j.issn.1006-267x.2022.05.059

Abstract

The purpose of this study was to screen and develop lactic acid bacteria for the preparation of probiotics for livestock and poultry, and explore the probiotic properties of L.crispatus Y68. Its probiotic properties were evaluated by antibacterial activity screening, growth curve measurement, acid production, drug sensitivity test, in vitro simulated gastrointestinal condition tolerance test, self-agglutination test and hydrophobicity test. The results showed that L.crispatus Y68 had strong inhibitory effect on Salmonella pullorum ATCC19945 and Staphylococcus aureus USA300, with diameter of bacteriostatic zone of (21.64±0.31) mm and (15.13±0.23) mm, respectively; the logarithmic growth period of L.crispatus Y68 was 6 to 24 h; after 48 h of culture, the pH of the cell suspension was stable at 3.89; L.crispatus Y68 was sensitive to erythromycin, penicillin, cephalosporin, chloramphenicol, vancomycin and clindamycin, but resistant to norfloxacin, ofloxacin, tetracycline, gentamicin and furazolidone; the survival rate of L.crispatus Y68 was 52.61% at pH 2.0 for 3 h, 65.92% at artificial gastric juice for 3 h, 86.23% at artificial intestinal juice for 10 h, and which could tolerate 0.3% bile salt;the self-aggregation rate of L.crispatus Y68 at 2 and 24 h were respectively(29.71±0.40)% and (97.23±0.76)%, and the hydrophobic percentage was (89.59±0.63)%. In conclusion, L.crispatus Y68 is tolerant toward simulated gastrointestinal tract conditions, including low pH, pepsin, pancreatin and bile salts, maintains strong self-aggregation ability and hydrophobic performance. Thus, L.crispatus Y68 from chickens is a potential candidate strain of livestock and poultry probiotics,applied to nutrition and health of livestock and poultry.

参考文献

[1] BARRIENTOS-DURÁN A, FUENTES-LÓPEZ A, DE SALAZAR A, et al.Reviewing the composition of vaginal microbiota:inclusion of nutrition and probiotic factors in the maintenance of eubiosis[J].Nutrients, 2020, 12(2):419.
[2] HEINTZ-BUSCHART A, WILMES P.Human gut microbiome:function matters[J].Trends in Microbiology, 2018, 26(7):563-574.  
[3] NEVELING D P, DICKS L M T.Probiotics:an antibiotic replacement strategy for healthy broilers and productive rearing[J].Probiotics and Antimicrobial Proteins, 2021, 13(1):1-11.  
[4] SCHIFFRIN E J, BLUM S.Interactions between the microbiota and the intestinal mucosa[J].European Journal of Clinical Nutrition, 2002, 56(Suppl.3):S60-S64.
[5] AZAD M A K, SARKER M, LI T J, et al.Probiotic species in the modulation of gut microbiota:an overview[J].BioMed Research International, 2018, 2018:9478630.
[6] AMARA A A, SHIBL A.Role of probiotics in health improvement, infection control and disease treatment and management[J].Saudi Pharmaceutical Journal, 2015, 23(2):107-114.  
[7] MASUMIZU Y, ZHOU B H, KOBER A K M H, et al.Isolation and immunocharacterization of Lactobacillus salivarius from the intestine of wakame-fed pigs to develop novel"immunosynbiotics"[J].Microorganisms, 2019, 7(6):167.
[8] 郑昕, 宫利宏, 唐霄, 等.鸡源肠道乳酸菌的筛选及抑菌作用分析[J].饲料研究, 2020, 43(10):56-59. ZHENG X, GONG L H, TANG X, et al.Screening of Lactobacillus from chickens intestine and analysis of bacteriostasis[J].Feed Research, 2020, 43(10):56-59.(in Chinese)
[9] 周子吕, 古绍彬, 吴影, 等.凝结芽孢杆菌CGMCC 9951抑菌特性研究[J].河南科技大学学报(自然科学版), 2021, 42(4):77-82, 9. ZHOU Z L, GU S B, WU Y, et al.Studies of antibacterial properties of Bacillus coagulans CGMCC 9951[J].Journal of Henan University of Science & Technology (Natural Science), 2021, 42(4):77-82, 9.(in Chinese)
[10] PACE N R.Mapping the tree of life:progress and prospects[J].Microbiology and Molecular Biology Reviews, 2009, 73(4):565-576.  
[11] CHARTERIS W P, KELLY P M, MORELLI L, et al.Antibiotic susceptibility of potentially probiotic Lactobacillus species[J].Journal of Food Protection, 1998, 61(12):1636-1643.  
[12] AHIRE J J, JAKKAMSETTY C, KASHIKAR M S, et al.In vitro evaluation of probiotic properties of Lactobacillus plantarum UBLP40 isolated from traditional indigenous fermented food[J].Probiotics and Antimicrobial Proteins, 2021, 13(5):1413-1424.  
[13] SORNSENEE P, SINGKHAMANAN K, SANGKHATHAT S, et al.Probiotic properties of Lactobacillus species isolated from fermented palm sap in Thailand[J].Probiotics and Antimicrobial Proteins, 2021, 13(4):957-969.  
[14] CHEN C C, LAI C C, HUANG H L, et al.Antimicrobial activity of Lactobacillus species against carbapenem-resistant Enterobacteriaceae[J].Frontiers in Microbiology, 2019, 10:789.
[15] DEL RE B, SGORBATI B, MIGLIOLI M, et al.Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum[J].Letters in Applied Microbiology, 2000, 31(6):438-442.  
[16] TERAI T, KATO K, ISHIKAWA E, et al.Safety assessment of the candidate oral probiotic Lactobacillus crispatus YIT 12319:analysis of antibiotic resistance and virulence-associated genes[J].Food and Chemical Toxicology, 2020, 140:111278.
[17] ZHANG Q X, ZHANG L L, ROSS P, et al.Comparative genomics of Lactobacillus crispatus from the gut and vagina reveals genetic diversity and lifestyle adaptation[J].Genes, 2020, 11(4):360.
[18] ESLAMI S, HADJATI J, MOTEVASELI E, et al.Lactobacillus crispatus strain SJ-3C-US induces human dendritic cells (DCs) maturation and confers an anti-inflammatory phenotype to DCs[J].APMIS, 2016, 124(8):697-710.  
[19] TOBITA K, YANAKA H, OTANI H.Anti-allergic effects of Lactobacillus crispatus KT-11 strain on ovalbumin-sensitized BALB/c mice[J].Animal Science Journal, 2010, 81(6):699-705.  
[20] AZAM R, GHAFOURI-FARD S, TABRIZI M, et al.Lactobacillus acidophilus and Lactobacillus crispatus culture supernatants downregulate expression of cancer-testis genes in the MDA-MB-231 cell line[J].Asian Pacific Journal of Cancer Prevention, 2014, 15(10):4255-4259.  
[21] SCILLATO M, SPITALE A, MONGELLI G, et al.Antimicrobial properties of Lactobacillus cell-free supernatants against multidrug-resistant urogenital pathogens[J].MicrobiologyOpen, 2021, 10(2):e1173.
[22] WANG S, WANG Q Y, YANG E C, et al.Antimicrobial compounds produced by vaginal Lactobacillus crispatus are able to strongly inhibit Candida albicans growth, hyphal formation and regulate virulence-related gene expressions[J].Frontiers in Microbiology, 2017, 8:564.
[23] LEE Y, SEO H, KIM S, et al.Activity of Lactobacillus crispatus isolated from vaginal microbiota against Mycobacterium tuberculosis[J].Journal of Microbiology, 2021, 59(11):1019-1030.  
[24] AMMOR M S, GUEIMONDE M, DANIELSEN M, et al.Two different tetracycline resistance mechanisms, plasmid-carried tet(L) and chromosomally located transposon-associated tet(M), coexist in Lactobacillus sakei Rits 9[J].Applied and Environmental Microbiology, 2008, 74(5):1394-1401.  
[25] NAMI Y, VASEGHI BAKHSHAYESH R, MOHAMMADZADEH JALALY H, et al.Probiotic properties of Enterococcus isolated from artisanal dairy products[J].Frontiers in Microbiology, 2019, 10:300.
[26] GÓMEZ ZAVAGLIA A, KOCIUBINSKI G, PÉREZ P, et al.Isolation and characterization of Bifidobacterium strains for probiotic formulation[J].Journal of Food Protection, 1998, 61(7):865-873.  
[27] SERVIN A L, COCONNIER M H.Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens[J].Best Practice & Research.Clinical Gastroenterology, 2003, 17(5):741-754.  
[28] SHIVANGI S, DEVI P B, RAGUL K, et al.Probiotic potential of Bacillus strains isolated from an acidic fermented food idli[J].Probiotics Antimicro, 2020, 12(4):1502-1513.  
[29] PINO A, BARTOLO E, CAGGIA C, et al.Detection of vaginal Lactobacilli as probiotic candidates[J].Scientific Reports, 2019, 9(1):3355.
[30] CESENA C, MORELLI L, ALANDER M, et al.Lactobacillus crispatus and its nonaggregating mutant in human colonization trials[J].Journal of Dairy Science, 2001, 84(5):1001-1010.  
[31] VOLTAN S, CASTAGLIUOLO I, ELLI M, et al.Aggregating phenotype in Lactobacillus crispatus determines intestinal colonization and TLR2 and TLR4 modulation in murine colonic mucosa[J].Clinical and Vaccine Immunology, 2007, 14(9):1138-1148.  
文章导航

/