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使用牛子宫阜上皮细胞模型对牛源分离株进行特性分析。

Characterisation of isolates from cattle using a bovine caruncular epithelial cell model.

作者信息

Blanchard Adam M, Billenness Rosemarie, Warren Jessica, Glanvill Amy, Roden William, Drinkall Emma, Maboni Grazieli, Robinson Robert S, Rees Catherine E D, Pfarrer Christiane, Tötemeyer Sabine

机构信息

School of Veterinary Medicine and Science, University of Nottingham, Nottingham, UK.

Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.

出版信息

Heliyon. 2020 Jul 23;6(7):e04476. doi: 10.1016/j.heliyon.2020.e04476. eCollection 2020 Jul.

Abstract

is an important foodborne pathogen in human and veterinary health, causing significant morbidity and mortality including abortion. It has a particular tropism for the gravid uterus, however, the route of infection in reproductive tissues of ruminants (i.e. placentome), is much less clear. In this study, we aimed to investigate a bovine caruncular epithelial cell (BCEC) line as a model for infection of the bovine reproductive tract. The BCEC infection model was used to assess the ability of 14 different isolates to infect these cells. Lysozyme sensitivity and bacterial survival in 580 μg lysozyme/ml correlated with attenuated ability to proliferate in BCEC (p = 0.004 and p = 0.02, respectively). Four isolates were significantly attenuated compared to the control strain 10403S. One of these strains (AR008) showed evidence of compromised cell wall leading to increased sensitivity to ß-lactam antibiotics, and another (7644) had compromised cell membrane integrity leading to increased sensitivity to cationic peptides. Whole genome sequencing followed by Multi Locus Sequence Type analysis identified that five invasive isolates had the same sequence type, ST59, despite originating from three different clinical conditions. Virulence gene analysis showed that the attenuated isolate LM4 was lacking two virulence genes (, ) known to be involved in intracellular growth and virulence. In conclusion, the BCEC model was able to differentiate between the infective potential of different isolates. Moreover, resistance to lysozyme correlated with the ability to invade and replicate within BCEC, suggesting co-selection for surviving challenging environments as the abomasum.

摘要

是人类和兽医健康领域中一种重要的食源性病原体,可导致包括流产在内的显著发病率和死亡率。它对妊娠子宫具有特殊的嗜性,然而,反刍动物生殖组织(即胎盘叶)的感染途径尚不清楚得多。在本研究中,我们旨在研究一种牛肉阜上皮细胞(BCEC)系作为牛生殖道感染的模型。BCEC感染模型用于评估14种不同分离株感染这些细胞的能力。溶菌酶敏感性和在580μg溶菌酶/ml中的细菌存活与在BCEC中增殖能力减弱相关(分别为p = 0.004和p = 0.02)。与对照菌株10403S相比,有4种分离株显著减弱。其中一种菌株(AR008)显示出细胞壁受损的证据,导致对β-内酰胺抗生素敏感性增加,另一种(7644)细胞膜完整性受损,导致对阳离子肽敏感性增加。全基因组测序随后进行多位点序列类型分析确定,尽管5种侵袭性分离株源自三种不同临床情况,但它们具有相同的序列类型ST59。毒力基因分析表明,减毒分离株LM4缺少两个已知参与细胞内生长和毒力的毒力基因(,)。总之,BCEC模型能够区分不同分离株的感染潜力。此外,对溶菌酶的抗性与在BCEC内侵袭和复制的能力相关,表明在作为皱胃的具有挑战性环境中生存存在共同选择。

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