奶牛阴道乳杆菌抗菌物质的体外特性
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摘要
采用牛津杯法测定4株乳杆菌(罗伊乳杆菌、格氏乳杆菌、约氏乳杆菌和唾液乳杆菌)对致病菌的抑菌活性,使用含有3,3,5,5-四甲基联苯胺和辣根过氧化物酶的MRS琼脂培养基测定乳杆菌产过氧化氢能力,最后通过蛋白类排除试验、抑菌物质热稳定性试验、有机酸排除试验和过氧化氢(H_2O_2)排除试验分析4株乳杆菌主要的抗菌物质。结果显示:4株菌株均具有良好的抗菌活性,约氏乳杆菌和唾液乳杆菌能够产生H_2O_2;约氏乳杆菌主要的抗菌物质为有机酸,罗伊乳杆菌、格氏乳杆菌和唾液乳杆菌主要抗菌物质除了有机酸外,仍可以产生细菌素。结果表明:罗伊乳杆菌、格氏乳杆菌和唾液乳杆菌可以作为防治奶牛子宫内膜炎的益生菌使用。
Analysis of the major anti-bacteria substance of Lactobacillus from healthy dairy cow vaginas to provide new strains for prevention and treatment of dairy cow endometritis.4 Lactobacilli strains( Lactobacillus reuteri,Lactobacillus gasseri,Lactobacillus johnsonii and Lactobacillus salivarius) were evaluated for anti-bacteria activity by Oxford cup method and test their ability to produce hydrogen peroxide by MRS agar medium with 3,3,5,5-tetramethyl benzidine and horseradish peroxidase. Finally, analysis of the major antibacterial substances was performed through excluding protease inhibition, heat stability test, hydrogen peroxide inhibition and organic acid inhibition.The results show that 4 strains have good antibacterial activities.Lactobacillus johnsonii and Lactobacillus salivarius can produce H_2O_2. The major antibacterial substance of Lactobacillus johnsonii is organic acid.Lactobacillus reuteri, Lactobacillus gasseri and Lactobacillus salivarius except organic acid also can produce bacteriocin. Lactobacillus reuteri, Lactobacillus gasseri and Lactobacillus salivarius can be used as probiotics to prevent and treat dairy cow endometritis.
Analysis of the major anti-bacteria substance of Lactobacillus from healthy dairy cow vaginas to provide new strains for prevention and treatment of dairy cow endometritis.4 Lactobacilli strains( Lactobacillus reuteri,Lactobacillus gasseri,Lactobacillus johnsonii and Lactobacillus salivarius) were evaluated for anti-bacteria activity by Oxford cup method and test their ability to produce hydrogen peroxide by MRS agar medium with 3,3,5,5-tetramethyl benzidine and horseradish peroxidase. Finally, analysis of the major antibacterial substances was performed through excluding protease inhibition, heat stability test, hydrogen peroxide inhibition and organic acid inhibition.The results show that 4 strains have good antibacterial activities.Lactobacillus johnsonii and Lactobacillus salivarius can produce H_2O_2. The major antibacterial substance of Lactobacillus johnsonii is organic acid.Lactobacillus reuteri, Lactobacillus gasseri and Lactobacillus salivarius except organic acid also can produce bacteriocin. Lactobacillus reuteri, Lactobacillus gasseri and Lactobacillus salivarius can be used as probiotics to prevent and treat dairy cow endometritis.
引文
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[4]HUANG R,TAO X,WAN C,et al.In vitro probiotic characteristics of Lactobacillus plantarum ZDY 2013and its modulatory effect on gut microbiota of mice[J].J Dairy Sci,2015,98(9):5850-5861.
[5]BORRIELLO S P,HAMMES W P,HOLZAPFEL W,et al.Safety of probiotics that contain Lactobacilli or Bifidobacteria[J].Clin Infect Dis,2003,36(6):775-780.
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[7]KAEWNOPPARAT S,DANGMANEE N,KAEWNOPPARAT N,et al.In vitro probiotic properties of Lactobacillus fermentum SK5isolated from vagina of a healthy woman[J].Anaerobe,2013,22:6-13.
[8]PAROLIN C,MARANGONI A,LAGHI L,et al.Isolation of vaginal lactobacilli and characterization of anti-Candida activity[J].PLoS One,2015,10(6):e0131220.
[9]OTERO M C,NADER-MACIAS M E.Inhibition of Staphylococcusaureusby H2O2-producing Lactobacillus gasseri isolated from the vaginal tract of cattle[J].Anim Reprod Sci,2006,96(1/2):35-46.
[10]DIMITONOVA S P,DANOVA S T,SERKEDJIEVA J P,et al.Antimicrobial activity and protective properties of vaginal Lactobacilli from healthy Bulgarian women[J].Anaerobe,2007,13(5/6):178-184.
[11]MALDONADO N C,DE RUIZ C S,OTERO M C,et al.Lactic acid bacteria isolated from young calves--characterization and potential as probiotics[J].Res Vet Sci,2012,92(2):342-349.
[12]MATU M N,ORINDA G O,NJAGI E N,et al.In vitro inhibitory activity of human vaginal Lactobacilli against pathogenic bacteria associated with bacterial vaginosis in Kenyan women[J].Anaerobe,2010,16(3):210-215.
[13]ANNUK H,SHCHEPETOVA J,KULLISAAR T,et al.Characterization of intestinal Lactobacilli as putative probiotic candidates[J].J Appl Microbiol,2003,94(3):403-412.
[14]LEE Y.Characterization of Weissella kimchii PL9023as a potential probiotic for women[J].FEMS Microbiol Lett,2005,250(1):157-162.
[2]COHEN C R,MANHART L E,BUKUSI E A,et al.Association between Mycoplasma genitalium and acute endometritis[J].Lancet(London,England),2002,359(9308):765-766.
[3]WANG Y,AMETAJ B N,AMBROSE D J,et al.Characterisation of the bacterial microbiota of the vagina of dairy cows and isolation of pediocin-producing Pediococcus acidilactici[J].BMC Microbiol,2013,13:19.
[4]HUANG R,TAO X,WAN C,et al.In vitro probiotic characteristics of Lactobacillus plantarum ZDY 2013and its modulatory effect on gut microbiota of mice[J].J Dairy Sci,2015,98(9):5850-5861.
[5]BORRIELLO S P,HAMMES W P,HOLZAPFEL W,et al.Safety of probiotics that contain Lactobacilli or Bifidobacteria[J].Clin Infect Dis,2003,36(6):775-780.
[6]JANKOVIC I,SYBESMA W,PHOTHIRATH P,et al.Application of probiotics in food products--challenges and new approaches[J].Curr Opin Biotechnol,2010,21(2):175-181.
[7]KAEWNOPPARAT S,DANGMANEE N,KAEWNOPPARAT N,et al.In vitro probiotic properties of Lactobacillus fermentum SK5isolated from vagina of a healthy woman[J].Anaerobe,2013,22:6-13.
[8]PAROLIN C,MARANGONI A,LAGHI L,et al.Isolation of vaginal lactobacilli and characterization of anti-Candida activity[J].PLoS One,2015,10(6):e0131220.
[9]OTERO M C,NADER-MACIAS M E.Inhibition of Staphylococcusaureusby H2O2-producing Lactobacillus gasseri isolated from the vaginal tract of cattle[J].Anim Reprod Sci,2006,96(1/2):35-46.
[10]DIMITONOVA S P,DANOVA S T,SERKEDJIEVA J P,et al.Antimicrobial activity and protective properties of vaginal Lactobacilli from healthy Bulgarian women[J].Anaerobe,2007,13(5/6):178-184.
[11]MALDONADO N C,DE RUIZ C S,OTERO M C,et al.Lactic acid bacteria isolated from young calves--characterization and potential as probiotics[J].Res Vet Sci,2012,92(2):342-349.
[12]MATU M N,ORINDA G O,NJAGI E N,et al.In vitro inhibitory activity of human vaginal Lactobacilli against pathogenic bacteria associated with bacterial vaginosis in Kenyan women[J].Anaerobe,2010,16(3):210-215.
[13]ANNUK H,SHCHEPETOVA J,KULLISAAR T,et al.Characterization of intestinal Lactobacilli as putative probiotic candidates[J].J Appl Microbiol,2003,94(3):403-412.
[14]LEE Y.Characterization of Weissella kimchii PL9023as a potential probiotic for women[J].FEMS Microbiol Lett,2005,250(1):157-162.
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