Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico-Quimicas y Naturales, Universidad Nacional de Río Cuarto, Ruta Nacional 36, Km 601, X5804ZAB Río Cuarto, Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico-Quimicas y Naturales, Universidad Nacional de Río Cuarto, Ruta Nacional 36, Km 601, X5804ZAB Río Cuarto, Córdoba, Argentina.
Res Vet Sci. 2020 Oct;132:33-41. doi: 10.1016/j.rvsc.2020.05.016. Epub 2020 May 20.
Streptococcus uberis is one of the most prevalent environmental pathogens of bovine mastitis. Biofilm growth ability by S. uberis looks to depend first upon the adherence of cells to a surface. The S. uberis ability to adhere to mammary gland epithelia might provide an advantage to colonize the lactating mammary gland. The objectives of this study were (a) to select S.uberis strains according to their ability to form biofilm, (b) to determine adherence to and internalization into MAC-T cells and (c) to investigate the expression profile adherence genes in these S. uberis strains. For the assays, the MAC-T bovine mammary epithelial cell line was used. Relative expression of genes acdA, lmb, scpA, sua, fbp and lbp was quantified by RT-qPCR. We observed that the RC38 strain from clinical bovine mastitis showed in the six genes higher values than control in both conditions. While the strain with greater ability to adhere, from clinical mastitis and biofilm producer (RC29) evidenced higher values in group 1 (G) (bacteria after the initial contact with MAC-T cells) and decrease in group 2 (G) (both adhered and internalized bacteria) than control. Strains with a moderate or strong capacity for biofilm production showed significantly lower relative expression values in the G. In all adherence associated genes, strain RC19 showed relative expression values incremented in G while in G decreased expression. In conclusion, we did not find a single profile of relative expression because the relative expression levels of each gene differed depending on the strain and the co-culture stage of S. uberis cells from which RNA was obtained.
停乳链球菌是引起奶牛乳腺炎的最常见的环境病原体之一。停乳链球菌的生物膜生长能力似乎首先依赖于细胞对表面的附着。停乳链球菌对乳腺上皮细胞的黏附能力可能使其具有定植泌乳乳腺的优势。本研究的目的是:(a)根据生物膜形成能力选择停乳链球菌株;(b)测定其对 MAC-T 细胞的黏附和内化;(c)研究这些停乳链球菌株中黏附相关基因的表达谱。为此,我们使用了 MAC-T 牛乳腺上皮细胞系。采用 RT-qPCR 定量分析 acdA、lmb、scpA、sua、fbp 和 lbp 基因的相对表达。我们发现,来自临床乳腺炎的 RC38 株在两种条件下的这 6 个基因的相对表达值均高于对照。而黏附能力更强的、来自临床乳腺炎和生物膜生产者(RC29)的菌株在第 1 组(G)(细菌与 MAC-T 细胞最初接触后)的相对表达值高于对照组,而在第 2 组(G)(黏附的和内化的细菌)则低于对照组。具有中度或强生物膜生产能力的菌株在 G 中表现出显著较低的相对表达值。在所有与黏附相关的基因中,RC19 株在 G 中表现出相对表达值增加,而在 G 中表达值下降。总之,我们没有发现单一的相对表达谱,因为每个基因的相对表达水平取决于菌株以及获得 RNA 的停乳链球菌细胞的共培养阶段。