2株分离乳酸菌对奶牛阴道上皮细胞的黏附作用
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摘要
益生乳酸菌对奶牛阴道上皮细胞黏附能力的强弱是其发挥益生作用的前提条件,因此黏附能力是筛选益生菌的重要标准之一。为筛选出可以制备微生态制剂的益生乳酸菌菌株,本试验采用碳氢化合物黏着法、紫外分光光度法、酶联免疫吸附法,测定了2株乳酸菌SQ0048、SQ0054及混合菌株的疏水性能、自凝集能力、生物被膜形成能力及其影响因素,并采用瑞士吉姆萨染色法研究了2株乳酸菌及其混合菌株对奶牛阴道上皮细胞的黏附作用。结果显示,菌株SQ0048以及混合菌株在不同pH值条件下对正十二烷的疏水作用极显著性高于二甲苯(P<0.01);在3h时,菌株SQ0048在pH5时的自凝集能力极显著高于pH7和pH6时的自凝集能力(P<0.01);菌株SQ0048在pH5时的成膜能力最强,为强被膜生物形成株,菌株SQ0054为弱被膜生物形成株,混合菌株也为弱被膜生物形成株;混合菌株对奶牛阴道上皮细胞的黏附性极显著高于2株单一乳酸菌(P<0.01),2株单一乳酸菌对奶牛阴道上皮细胞的黏附具有协同作用。结果表明,2株乳酸菌及其混合菌株均具有良好的黏附能力,而混合菌株更适合用于微生态制剂的研发。
Adhesion ability of probiotic bacteria to epithelial cells from cow reproductive tract is the precondition to play aprobiotic role,therefore,adhesion ability is one of the important standards of probiotics for screening.In order to screen probiotics strains that can be used for microecological preparations,the hydrophobicity,self-agglutination ability,biofilm formation ability and influencing factors of two lactic acid bacteria SQ0048,SQ0054 and mixed strains were tested using the hydrocarbon adhesion method,ultraviolet spectrophotometry and enzyme-linked immunosorbent assay.Then,two strains of Lactobacillus and mixed strains with cow vaginal epithelial cells were employed to investigate the bacterial adhesion ability using Swiss Jimsa staining method.The results showed that the hydrophobicity of SQ0048 and mixed strains to dodecane was higher than that of xylene(P<0.01)at different pH conditions.At 3 h,self-agglutination ability of SQ0048 was significant at pH5(P<0.01).SQ0048 had the strongest ability of membrane formation at pH5,which was a strongly biofilm-forming strain.SQ0054 is a weak envelope organism;mixed strain is also a weak envelope organism.The adhesion of mixed strains to cow vaginal epithelial cells was significantly higher than that of two single lactic acid bacteria(P<0.01),and the two single lactic acid bacteria had a synergistic effect on the adhesion of cow vaginal epithelial cells.In conclusion,the two single strain and mixed strains have good adhesion ability,and the mixed strains are more suitable for the development of probiotics.
Adhesion ability of probiotic bacteria to epithelial cells from cow reproductive tract is the precondition to play aprobiotic role,therefore,adhesion ability is one of the important standards of probiotics for screening.In order to screen probiotics strains that can be used for microecological preparations,the hydrophobicity,self-agglutination ability,biofilm formation ability and influencing factors of two lactic acid bacteria SQ0048,SQ0054 and mixed strains were tested using the hydrocarbon adhesion method,ultraviolet spectrophotometry and enzyme-linked immunosorbent assay.Then,two strains of Lactobacillus and mixed strains with cow vaginal epithelial cells were employed to investigate the bacterial adhesion ability using Swiss Jimsa staining method.The results showed that the hydrophobicity of SQ0048 and mixed strains to dodecane was higher than that of xylene(P<0.01)at different pH conditions.At 3 h,self-agglutination ability of SQ0048 was significant at pH5(P<0.01).SQ0048 had the strongest ability of membrane formation at pH5,which was a strongly biofilm-forming strain.SQ0054 is a weak envelope organism;mixed strain is also a weak envelope organism.The adhesion of mixed strains to cow vaginal epithelial cells was significantly higher than that of two single lactic acid bacteria(P<0.01),and the two single lactic acid bacteria had a synergistic effect on the adhesion of cow vaginal epithelial cells.In conclusion,the two single strain and mixed strains have good adhesion ability,and the mixed strains are more suitable for the development of probiotics.
引文
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[9]ZELJKA PAVELI C,SKALKO-BASNET N,JALSENJAK I.Liposomes containing drugs for treatment of vaginal infections[J].Eur J Pharmaceut Sci,1999,8(4):345-351.
[10] MARIANI C,OULAHAL N,CHAMBA J F,et al.Inhibition of Listeria monocytogenes by resident biofilms present on wooden shelves used for cheese ripening[J].Food Control,2011,22(8):1357-1362.
[2]BLUM S,RENIERO R,SCHIFFRIN E J,et al.Adhesion studies for probiotics:need for calidation and refinement[J].Trends Food Sci Technol,1999,21(10):405-410.
[3]NAITO Y,TOHDA H,OKUDA K,et al.Adherence and hydrophobicity of invasion and noninvasive strains of porphyromoonas gingivalis[J].Oral Microbiol Immunol,1993,8(4):195.
[4]邵明旭,刘丽萍,于翠莲,等.具益生特性的小尾寒羊肠道正常菌群的分离、鉴定及筛选[J].中国兽医学报,2016,36(12):2054-2060.
[5]HARBOE M,OETTINGER T,WIKER H G,et al.Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG[J].Infect Immun,1996,64(1):16.
[6]DEL RE B,SGORBATI B,MIGLIOLI M,et al.Adhesion,autoaggrega-tion and hydrophobicity of 13strains of Bifidobacterium longum[J].Lett Appl Microbiol,2000,31:438-442.
[7]PAN W H,LI P L,LIU Z.The correlation between surface hydrophobicity and adherence of Bifidobacterium strains from centenariansfaeces[J].Anaerobe,2006,12(3):148-152.
[8]RODRIGUEZ C,COFRE J V,SANCHEZ M,et al.Lactobacilli isolated from vaginal vault of dairy and meat cows during progesteronic stage of estrous cycle[J].Anaerobe,2011,17(1):15-18.
[9]ZELJKA PAVELI C,SKALKO-BASNET N,JALSENJAK I.Liposomes containing drugs for treatment of vaginal infections[J].Eur J Pharmaceut Sci,1999,8(4):345-351.
[10] MARIANI C,OULAHAL N,CHAMBA J F,et al.Inhibition of Listeria monocytogenes by resident biofilms present on wooden shelves used for cheese ripening[J].Food Control,2011,22(8):1357-1362.