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Restoring catalase activity in Staphylococcus aureus subsp. anaerobius leads to loss of pathogenicity for lambs

机译:恢复金黄色葡萄球菌亚种中的过氧化氢酶活性。厌食导致羔羊致病性丧失

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摘要

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.
机译:金黄色葡萄球菌亚种。厌氧杆菌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,脓肿是绵羊和山羊的一种特殊的慢性疾病,其特征是形成坏死性病变,通常位于浅表淋巴结。我们构建了一个金黄色葡萄球菌亚种的同基因突变体。 anaerobius(RDKA84)携带来自金黄色葡萄球菌ATCC 12600的经过修复的功能性过氧化氢酶基因,以调查金黄色葡萄球菌亚种中是否缺乏过氧化氢酶。厌氧菌在其生理和致病特性中起作用。过氧化氢酶活性对RDKA84的体外生长特性没有明显的影响,RDKA84与野生型一样,在需氧培养的琼脂平板上也不生长。在死亡分析中进行分析时,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中性粒细胞(PMN)中过氧化氢酶阳性突变体RDKA84的细胞内存活率显着高于野生型,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,未观察到细胞内存活率的差异。有趣的是,与野生型金黄色葡萄球菌亚种相比,过氧化氢酶阳性突变体RDKA84对羔羊(脓肿疾病的天然宿主)的致病力大大降低。厌氧菌感染的两个实验模型。

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