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实验性奶牛乳腺炎感染后,两种适应基因型不同的金黄色葡萄球菌菌株引起的免疫反应差异。

Differential immune response to two Staphylococcus aureus strains with distinct adaptation genotypes after experimental intramammary infection of dairy cows.

机构信息

Laboratorio de Biología Celular y Molecular Aplicada, Facultad de Ciencias Veterinarias, Universidad Nacional del Litoral, Argentina; Instituto de Ciencias Veterinarias del Litoral (ICIVET-Litoral), Universidad Nacional del Litoral - Consejo Nacional de Investigaciones Científicas y Tecnológicas, (UNL-CONICET), Argentina.

Instituto de Investigación de la Cadena Láctea (INTA-CONICET), Estación Experimental Agropecuaria Rafaela, Ruta 34 Km 227, Rafaela, Santa Fe, Argentina.

出版信息

Microb Pathog. 2022 Nov;172:105789. doi: 10.1016/j.micpath.2022.105789. Epub 2022 Sep 20.

Abstract

The aim of this study was to evaluate and compare the ability of two S. aureus strains with different adaptation genotypes (low and high) to the bovine mammary gland (MG) to establish an intramammary infection (IMI) and induce an immune response after an experimental challenge in lactating cows. Two isolates (designated 806 and 5011) from bovine IMI with different genotypic profiles, harboring genes involved in adherence and biofilm production, belonging to different capsular polysaccharide (CP) type, accessory gene regulator (agr) group, pulsotype (PT) and sequence type/clonal complex (ST/CC) were selected. Strains 806 and 5011 were associated with low (nonpersistent-NP) and high (persistent-P) adaptation to the MG, respectively. Strain 806 (NP) was characterized as agr group II, cap5 positive and ST350; strain 5011 (P) agr group I, cap8 positive and CC188. Three groups of clinically healthy cows, 4 cows/treatment group, were inoculated by the intramammary route with strain 806 (NP), strain 5011 (P) and pyrogen-free saline solution. All mammary quarters challenged with strain 806 (NP) developed mild clinical mastitis between 1 and 7 d post inoculation (pi). Quarters challenged with strain 5011 (P) developed a persistent IMI; bacteria were recovered from milk from d 7 pi and up to d 56 pi. In quarters inoculated with strain 806 (NP) the inflammatory response induced was greater and earlier than the one induced by strain 5011 (P), since a somatic cell count (SCC) peak was observed at d 2 pi, while in quarters inoculated with strain 5011 (P) no variations in SCC were observed until d 4 pi reaching the maximum values at d 14 pi; indicating a lower and delayed initial inflammatory response. The highest levels of nitric oxide (NO) and lactoferrin (Lf) detected in milk from quarters inoculated with both S. aureus strains coincided with the highest SCC at the same time periods, indicating an association with the magnitude of inflammation. The high levels of IL-1β induced by strain 806 (NP) were associated with the highest SCC detected (d 2 pi); while quarters inoculated with strain 5011 (P) showed similar IL-1β levels to those found in control quarters. In quarters inoculated with strain 806 (NP) two peaks of IL-6 levels on d 2 and 14 pi were observed; while in quarters inoculated with strain 5011 (P) IL-6 levels were similar to those found in control quarters. The strain 806 (NP) induced a higher total IgG and IgG response; while strain 5011 (P) generated a higher IgG response (even against the heterologous strain). The present study demonstrated that S. aureus strains with different genotype and adaptability to bovine MG influence the local host immune response and the course and severity of the infectious process.

摘要

本研究旨在评估和比较两株适应基因型(低和高)不同的金黄色葡萄球菌(S. aureus)菌株进入牛乳腺(MG)建立乳腺炎(IMI)的能力,并在泌乳奶牛中进行实验性攻毒后诱导免疫反应。从牛乳腺炎中分离出两株(分别命名为 806 和 5011),具有不同的基因型特征,携带与粘附和生物膜产生相关的基因,分别属于不同的荚膜多糖(CP)型、辅助基因调节(agr)群、脉冲型(PT)和序列型/克隆复合体(ST/CC)。菌株 806 和 5011 分别与 MG 的低(非持续性-NP)和高(持续性-P)适应性相关。菌株 806(NP)的特征是 agr 组 II,cap5 阳性和 ST350;菌株 5011(P)agr 组 I,cap8 阳性和 CC188。三组临床健康奶牛,每组 4 头,通过乳腺内途径接种菌株 806(NP)、菌株 5011(P)和无热原盐水。所有接种菌株 806(NP)的乳腺象限在接种后 1 至 7 天内(pi)均发生轻度临床乳腺炎。接种菌株 5011(P)的象限发展为持续性 IMI;从 d 7 pi 开始并持续到 d 56 pi 从牛奶中回收细菌。接种菌株 806(NP)引起的炎症反应比接种菌株 5011(P)引起的炎症反应更大、更早,因为在 d 2 pi 时观察到体细胞计数(SCC)峰值,而接种菌株 5011(P)的象限在 d 4 pi 之前没有 SCC 变化,直到 d 14 pi 达到最大值;表明初始炎症反应较低且延迟。在接种两株 S. aureus 菌株的象限中检测到的最高水平的一氧化氮(NO)和乳铁蛋白(Lf)与同一时期的最高 SCC 同时出现,表明与炎症程度有关。菌株 806(NP)诱导的高水平 IL-1β与检测到的最高 SCC 相关(d 2 pi);而接种菌株 5011(P)的象限与对照象限中发现的 IL-1β 水平相似。在接种菌株 806(NP)的象限中观察到两个 IL-6 水平的高峰,分别在 d 2 和 14 pi;而接种菌株 5011(P)的象限中 IL-6 水平与对照象限相似。菌株 806(NP)诱导了更高的总 IgG 和 IgG 反应;而菌株 5011(P)产生了更高的 IgG 反应(甚至针对异源菌株)。本研究表明,具有不同基因型和适应牛 MG 能力的金黄色葡萄球菌菌株影响局部宿主免疫反应以及感染过程的进程和严重程度。

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