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从中俄罗斯临床型隐性乳房炎奶牛乳中分离的金黄色葡萄球菌的外毒素多样性。

Exotoxin diversity of Staphylococcus aureus isolated from milk of cows with subclinical mastitis in Central Russia.

机构信息

L. K. Ernst All-Russian Research Institute of Animal Husbandry, Federal Agency of Scientific Organizations, Dubrovitsy, 142132, Russia; Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Pushchino, 142290, Russia.

L. K. Ernst All-Russian Research Institute of Animal Husbandry, Federal Agency of Scientific Organizations, Dubrovitsy, 142132, Russia; Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Pushchino, 142290, Russia.

出版信息

J Dairy Sci. 2018 May;101(5):4325-4331. doi: 10.3168/jds.2017-14074. Epub 2018 Feb 21.

Abstract

Mastitis, a major veterinary problem widespread in many regions, is caused mainly by Staphylococcus spp. However, there is no current reliable information about the role of Staphylococcus aureus and their toxins in the development of mastitis in cows in the territory of the Russian Federation. The aim of this investigation was to determine the profile of exotoxins of S. aureus from cow milk from farms of Central Russia. A total of 60 isolates of S. aureus were obtained from milk samples of cows with the subclinical form of mastitis. The exotoxin genes were identified using 2 types of PCR assays. The diversity of enterotoxin genes was studied by multiplex PCR. The percentage occurrence of enterotoxin genes was as follows: sea, 53.3%; seb, 3.3%; sec, 50%; sed, 4%; see, 46.6%; seg, 70%; sei, 10%; selp, 3.3%; and tsst1, 1.6%. The seh gene was not detected. The genes of pore-forming toxins and phenol-soluble modulins were identified by singleplex PCR and consisted of the following: hlA, 70%; lucS, 46.6%; psmA, 81.6%; psmB, 95%; and hld, 78.3%. The most abundant genes were psm (psmB, 95%), which codes for pore-forming toxins, and seg (70%), which codes for enterotoxins. The production of some enterotoxins in bacterial culture medium was detected by ELISA. The level of toxin production was near 1 ng/mL for SEA, SEE, SEG, SEI, SELP, and TSST-1 and reached a maximal level of 18 ng/mL for SEE. In the present work, we show that subclinical mastitis in cows is associated with S. aureus in the central region of the Russian Federation. Most of the isolates containing enterotoxin genes also had cytotoxin genes.

摘要

乳腺炎是一种在许多地区广泛存在的主要兽医问题,主要由葡萄球菌属引起。然而,目前没有关于俄罗斯联邦领土上奶牛乳腺炎中金黄色葡萄球菌及其毒素作用的可靠信息。本研究旨在确定来自俄罗斯中部地区奶牛隐性乳腺炎牛奶中金黄色葡萄球菌的外毒素谱。从患有亚临床乳腺炎的奶牛牛奶样本中获得了总共 60 株金黄色葡萄球菌分离株。使用 2 种 PCR 检测法鉴定外毒素基因。通过多重 PCR 研究肠毒素基因的多样性。肠毒素基因的出现率如下:sea,53.3%;seb,3.3%;sec,50%;sed,4%;see,46.6%;seg,70%;sei,10%;selp,3.3%;和 tsst1,1.6%。未检测到 seh 基因。通过单重 PCR 鉴定了孔形成毒素和酚可溶性调节素基因,包括以下基因:hlA,70%;lucS,46.6%;psmA,81.6%;psmB,95%;和 hld,78.3%。最丰富的基因是 psm(psmB,95%),它编码孔形成毒素,以及 seg(70%),它编码肠毒素。通过 ELISA 在细菌培养基中检测到一些毒素的产生。毒素产生水平接近 SEA、SEE、SEG、SEI、SELP 和 TSST-1 的 1ng/mL,而 SEE 则达到 18ng/mL 的最大水平。在本工作中,我们表明,俄罗斯中部地区奶牛的亚临床乳腺炎与金黄色葡萄球菌有关。含有肠毒素基因的大多数分离株也含有细胞毒素基因。

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