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与泰国中部和东北部省份亚临床型牛乳腺炎相关的物种。

spp. associated with subclinical bovine mastitis in central and northeast provinces of Thailand.

作者信息

Pumipuntu Natapol, Tunyong Witawat, Chantratita Narisara, Diraphat Pornphan, Pumirat Pornpan, Sookrung Nitat, Chaicumpa Wanpen, Indrawattana Nitaya

机构信息

Department of Microbiology and Immunology/Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.

One Health Research Unit/Faculty of Veterinary Sciences, Maha Sarakham University, Maha Sarakham, Thailand.

出版信息

PeerJ. 2019 Mar 14;7:e6587. doi: 10.7717/peerj.6587. eCollection 2019.

Abstract

BACKGROUND

spp. are major cause of bovine mastitis (BM) worldwide leading to economic damage to dairy farms and public health threat. Recently, a newly emerged has been found as a human and animal pathogen. Molecular characteristics, virulence and antibiotic resistant phenotypes of bacteria causing BM in Thailand are rare. This study aimed to investigated spp. associated with subclinical bovine mastitis (SCM) in Thailand.

METHODS

Milk samples were collected from 224 cows of 52 dairy herds in four central and northeast provinces. Total somatic cell counts (SCC) and California mastitis test (CMT) were used to identify SCM cows. Milk samples were cultured for spp. Coagulase-positive isolates were subjected to pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Organisms suspected as were verified by detecting nonribosomal peptide synthetase gene. All isolates were checked for antibiograms and the presence of various virulence genes.

RESULTS

From the 224 milk samples of 224 cows, 132 (59%) were positive for SCM by SCC and CMT and 229 staphylococcal isolates were recovered. They were 32 coagulase-positive (24 and eight ) and 197 coagulase-negative. PFGE of the and revealed 11 clusters and a non-typeable pattern. MLST of representatives of the 11 PFGE clusters, three PFGE non-typeable isolates from different locations and showed 12 sequence types. The eight isolates belonged to ST1223 (three isolates), ST2250 (two isolates), and ST2793 (two isolates). The antimicrobial tests identified 11 (46%) methicillin-resistant and 25 (13%) methicillin-resistant coagulase-negative isolates, while seven were methicillin-susceptible and one isolate was methicillin-resistant. All of the 229 isolates were multiply resistant to other antibiotics. The most prevalent virulence genes of the 24 isolates were , and (X and IgG-binding region) (100%), (96%), (96%) and (79%). Six isolates carried one enterotoxin gene each and other virulence genes including and , indicating their pathogenic potential.

CONCLUSION AND PERSPECTIVE

This is the first report on the from cow milk samples with SCM. Data on the molecular characteristics, virulence genes and antibiograms of the spp. obtained from the present study showed a wide spread and increasing trend of methicillin-resistance and multiple resistance to other antibiotics. This suggests that the "One Health" practice should be nurtured, not only at the dairy farm level, but also at the national or even the international levels through cooperation of different sectors (dairy farmers, veterinarians, medical and public health personnel and scientists) in order to effectively combat and control the spread of these pathogens.

摘要

背景

葡萄球菌属是全球范围内引起奶牛乳房炎(BM)的主要原因,给奶牛场造成经济损失并对公共卫生构成威胁。最近,一种新出现的葡萄球菌已被发现是一种人畜共患病原体。泰国引起奶牛乳房炎的细菌的分子特征、毒力和抗生素耐药表型鲜有报道。本研究旨在调查泰国与亚临床奶牛乳房炎(SCM)相关的葡萄球菌属。

方法

从四个中部和东北部省份的52个奶牛场的224头奶牛中采集牛奶样本。使用总体细胞计数(SCC)和加利福尼亚乳房炎检测(CMT)来识别患SCM的奶牛。对牛奶样本进行葡萄球菌属培养。凝固酶阳性分离株进行脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)。通过检测非核糖体肽合成酶基因来验证疑似葡萄球菌的菌株。检查所有分离株的抗菌谱和各种毒力基因的存在情况。

结果

从224头奶牛的224份牛奶样本中,132份(59%)通过SCC和CMT检测为SCM阳性,并分离出229株葡萄球菌。其中32株为凝固酶阳性(24株金黄色葡萄球菌和8株其他葡萄球菌),197株为凝固酶阴性。金黄色葡萄球菌和其他葡萄球菌的PFGE显示出11个簇和1个不可分型模式。11个PFGE簇的代表菌株、来自不同地点的3株PFGE不可分型的其他葡萄球菌分离株以及金黄色葡萄球菌的MLST显示出12种序列类型。8株其他葡萄球菌分离株分别属于ST1223(3株)、ST2250(2株)和ST2793(2株)。抗菌试验鉴定出11株(46%)耐甲氧西林金黄色葡萄球菌和25株(13%)耐甲氧西林凝固酶阴性分离株,而7株金黄色葡萄球菌对甲氧西林敏感,1株分离株对甲氧西林耐药。所有229株分离株对其他抗生素均多重耐药。24株金黄色葡萄球菌分离株中最常见的毒力基因是sea、seb、sec(X和IgG结合区)(100%)、sdrC(96%)、fnbA(96%)和clfA(79%)。6株金黄色葡萄球菌分离株各携带一个肠毒素基因以及其他毒力基因,包括eta和etb,表明它们具有致病潜力。

结论与展望

这是关于来自患SCM奶牛牛奶样本中葡萄球菌属的首次报告。本研究获得的葡萄球菌属的分子特征、毒力基因和抗菌谱数据显示,耐甲氧西林和对其他抗生素多重耐药呈广泛传播且有增加趋势。这表明不仅应在奶牛场层面,而且应通过不同部门(奶农、兽医、医学和公共卫生人员以及科学家)的合作在国家甚至国际层面培育“同一健康”实践,以有效抗击和控制这些病原体的传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4f18/6421060/5a303d80cd5d/peerj-07-6587-g001.jpg

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