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考虑到乳杆菌在动物用益生菌制剂中的应用,对其凝集素样结合作用进行了研究。

Lectin-like binding of lactobacilli considered for their use in probiotical preparations for animal use.

作者信息

Styriak Igor, Nemcová Radomira

机构信息

Institute of Animal Physiology, Slovak Academy of Sciences, Kosice, Slovakia.

出版信息

Berl Munch Tierarztl Wochenschr. 2003 Mar-Apr;116(3-4):96-101.

Abstract

Three gut lactobacilli from piglets (Lactobacillus plantarum L 5, Lactobacillus paracasei L 81, Lactobacillus fermentum L 670) and Lactobacillus casei subsp. pseudoplantarum L.c.) from a calf were examined by microtitre plate binding assay for their lectin-like binding activity after their cultivation on Rogosa agar and in MRS broth. Three ECM (extracellular matrix) molecules (fetuin, porcine fibronectin and porcine mucin) were selected for this assay. Additionally, the effect of heparin on the binding of these three ECM molecules by Lactobacillus strains in microtitre plates was tested. Moreover, haemagglutination tests with pig, cattle, sheep, and hen erythrocytes were performed. However, none of the four Lactobacillus strains examined did react with any of the erythrocytes tested. The differences between individual strains were observed in their binding to immobilised ECM molecules. The best adherent was the Lactobacillus plantarum L5, however, the other three strains showed also good ECM binding. With regard to an influence of cultivation medium on lectin-like binding activity, binding of all ECM molecules was expressed in Lactobacillus paracasei L 81 to significantly higher degree (P < 0.001) after cultivation on Rogosa agar than in MRS broth. Similarly, strains Lactobacillus fermentum L 670 and Lactobacillus casei subsp. pseudoplantarum L.c. displayed significantly higher (P < 0.001) binding of fibronectin and mucin after growth on Rogosa agar in comparison with MRS broth cultivation. However, no significant (on fetuin and fibronectin binding) or opposite effect (on mucin binding) of cultivation medium was observed in Lactobacillus plantarum L 5 strain. The influence of cultivation medium on fetuin binding by Lactobacillus fermentum L 670 was also not significant while Lactobacillus casei subsp. pseudoplantarum L.c. bound fetuin significantly better (P < 0.01) after growth on Rogosa agar. Heparin pretreatment increased the binding of the ECM molecules by the Lactobacillus fermentum L 670 strain significantly (P < 0.001 or P < 0.05) with the exception of porcine fibronectin when the strain was cultivated in MRS broth. This result is important especially in the connection with the previous observations that heparin decreased ECM binding of enteropathogens as staphylococci or clinical enterococcal isolates. Following up on some earlier strain characteristics, these results indicate that the selected lactobacilli are probably suitable for probiotic purposes.

摘要

对来自仔猪的三种肠道乳酸杆菌(植物乳杆菌L5、副干酪乳杆菌L81、发酵乳杆菌L670)以及来自一头小牛的干酪乳杆菌假植物亚种L.c.,在罗戈萨琼脂培养基和MRS肉汤中培养后,通过微量滴定板结合试验检测它们的凝集素样结合活性。为此试验选择了三种细胞外基质(ECM)分子(胎球蛋白、猪纤连蛋白和猪粘蛋白)。此外,还检测了肝素对微量滴定板中这些乳酸杆菌菌株与这三种ECM分子结合的影响。而且,用猪、牛、羊和鸡的红细胞进行了血凝试验。然而,所检测的四种乳酸杆菌菌株均未与任何受试红细胞发生反应。在它们与固定化ECM分子的结合方面观察到了各菌株之间的差异。粘附性最佳的是植物乳杆菌L5,不过,其他三种菌株也表现出良好的ECM结合能力。关于培养基对凝集素样结合活性的影响,副干酪乳杆菌L81在罗戈萨琼脂培养基上培养后,与在MRS肉汤中培养相比,所有ECM分子的结合均显著更高(P < 0.001)。同样,发酵乳杆菌L670和干酪乳杆菌假植物亚种L.c.在罗戈萨琼脂培养基上生长后,与在MRS肉汤中培养相比,纤连蛋白和粘蛋白的结合显著更高(P < 0.001)。然而,在植物乳杆菌L5菌株中未观察到培养基对胎球蛋白和纤连蛋白结合有显著影响(或对粘蛋白结合有相反影响)。发酵乳杆菌L670培养基对胎球蛋白结合的影响也不显著,而干酪乳杆菌假植物亚种L.c.在罗戈萨琼脂培养基上生长后结合胎球蛋白的能力显著更强(P < 0.01)。肝素预处理显著增加了发酵乳杆菌L670菌株对ECM分子的结合(P < 0.001或P < 0.05),但该菌株在MRS肉汤中培养时对猪纤连蛋白的结合除外。这一结果尤其重要,因为之前的观察发现肝素会降低葡萄球菌或临床肠球菌分离株等肠道病原体与ECM的结合。基于一些早期菌株特性的后续研究,这些结果表明所选的乳酸杆菌可能适用于益生菌用途。

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