Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain.
Vet Res. 2010 Jul-Aug;41(4):41. doi: 10.1051/vetres/2010013. Epub 2010 Feb 22.
Staphylococcus aureus subsp. anaerobius, a microaerophilic and catalase-negative bacterium, is the etiological agent of abscess disease, a specific chronic condition of sheep and goats, which is characterized by formation of necrotic lesions that are located typically in superficial lymph nodes. We constructed an isogenic mutant of S. aureus subsp. anaerobius (RDKA84) that carried a repaired and functional catalase gene from S. aureus ATCC 12600, to investigate whether the lack of catalase in S. aureus subsp. anaerobius plays a role in its physiological and pathogenic characteristics. The catalase activity had no apparent influence on the in vitro growth characteristics of RDKA84, which, like the wild-type, did not grow on aerobically incubated agar plates. Restoration of catalase activity in RDKA84 substantially increased resistance to H2O2 when analyzed in a death assay. The intracellular survival rates of the catalase-positive mutant RDKA84 in polymorphonuclear neutrophils (PMN) isolated from adult sheep were significantly higher than those of the wild-type, while no differences were found with PMN isolated from lambs. RDKA84 showed significantly lower survival rates in murine macrophages (J774A.1 cells) than the wild-type strains did, whereas, in bovine mammary epithelial cells (MAC-T), no differences in intracellular survival were observed. Interestingly, the virulence for lambs, the natural host for abscess disease, of the catalase-positive mutant RDKA84 was reduced dramatically in comparison with wild-type S. aureus subsp. anaerobius in two experimental models of infection.
厌氧金黄色葡萄球菌亚种是一种微需氧和过氧化氢酶阴性的细菌,是脓肿病的病原体,这是绵羊和山羊特有的一种慢性疾病,其特征是形成坏死病变,通常位于浅表淋巴结中。我们构建了厌氧金黄色葡萄球菌亚种(RDKA84)的同源突变体,该突变体携带了来自金黄色葡萄球菌 ATCC 12600 的修复和功能正常的过氧化氢酶基因,以研究厌氧金黄色葡萄球菌亚种中过氧化氢酶的缺乏是否与其生理和致病特性有关。过氧化氢酶活性对 RDKA84 的体外生长特性没有明显影响,与野生型一样,它在有氧孵育的琼脂平板上不生长。在死亡测定中分析时,RDKA84 中过氧化氢酶活性的恢复大大提高了对 H2O2 的抗性。从成年绵羊分离的多形核白细胞(PMN)中,过氧化氢酶阳性突变体 RDKA84 的细胞内存活率明显高于野生型,而从羔羊分离的 PMN 则没有差异。RDKA84 在鼠巨噬细胞(J774A.1 细胞)中的存活率明显低于野生型菌株,而在牛乳腺上皮细胞(MAC-T)中则没有观察到细胞内存活的差异。有趣的是,与野生型厌氧金黄色葡萄球菌亚种相比,过氧化氢酶阳性突变体 RDKA84 在两种脓肿病感染的实验模型中对羔羊(脓肿病的天然宿主)的毒力显著降低。