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与山羊乳腺炎相关的对金黄色葡萄球菌具有毒性的噬菌体的分离、特性鉴定及治疗潜力评估

Isolation, characterization and therapeutic potential assessment of bacteriophages virulent to Staphylococcus aureus associated with goat mastitis.

作者信息

Mishra A K, Sharma N, Kumar A, Kumar N, Gundallahalli Bayyappa M R, Kumar S, Kumar N

机构信息

Goat Health Division, Central Institute for Research on Goats (CIRG), Makhdoom, Farah-281122, Mathura, Uttar Pradesh, India;

Project Directorate on Animal Disease Monitoring and Surveillance (PD-ADMAS), Hebbal-560024, Bengaluru, Karnataka, India;

出版信息

Iran J Vet Res. 2014 Fall;15(4):320-5.

Abstract

In the present study, the therapeutic potential of bacteriophages virulent to Staphylococcus aureus associated with goat mastitis were isolated, identified and assessed. Staphylococcus aureus (host or indicator bacterium) was isolated from a goat suffering from clinical mastitis. Based on cultural, morphological, biochemical tests and amplification of S. aureus specific thermonuclease gene in PCR, the identity of the organism was confirmed as S. aureus. Bacteriophages were isolated from soil and faecal samples (n=42) collected from different parts of the Mathura district in Uttar Pradesh (India), and their identity was confirmed by amplification of the bacteriophage-specific endolysin gene fragment in PCR. The thermal tolerance study revealed that all phage isolates were stable at 30 and 40°C with 100% lytic efficacy and their activities reduced to 62-80% at 50°C declining sharply at 60°C with less than 5% efficacy. Likewise, at pH = 6.5 and 7.5, the survivability of all isolates was 100% which reduced to 70-79% and 84-91% at pH = 5.5 and 8.5, respectively. All isolates were stable up to 3 months at 37°C, and for 16 months at 4°C but the stability of their respective endolysins only lasted for 12-23 days at 37°C and 6 months at 4°C. Three of the bacteriophage isolates, S. aureus phage/CIRG/1, S. aureus phage/CIRG/4 and S. aureus phage/CIRG/5 exhibited lytic activity against over 80% of the staphylococcal isolates. The results of the present study provide insight for the use of lytic bacteriophages for therapeutic interventions against multi-drug-resistant S. aureus inducing mastitis in goats.

摘要

在本研究中,对与山羊乳腺炎相关的、对金黄色葡萄球菌具有毒性的噬菌体的治疗潜力进行了分离、鉴定和评估。从患有临床乳腺炎的山羊中分离出金黄色葡萄球菌(宿主或指示菌)。基于培养、形态学、生化试验以及在聚合酶链反应(PCR)中对金黄色葡萄球菌特异性耐热核酸酶基因的扩增,确认该生物体为金黄色葡萄球菌。从印度北方邦马图拉地区不同地点采集的土壤和粪便样本(n = 42)中分离出噬菌体,并通过在PCR中扩增噬菌体特异性溶菌酶基因片段来确认其身份。耐热性研究表明,所有噬菌体分离株在30℃和40℃时均稳定,裂解效率为100%,在50℃时其活性降至62 - 80%,在60℃时急剧下降,效率低于5%。同样,在pH = 6.5和7.5时,所有分离株的存活率为100%,在pH = 5.5和8.5时分别降至70 - 79%和84 - 91%。所有分离株在37℃下可稳定保存3个月,在4℃下可稳定保存16个月,但其各自溶菌酶的稳定性在37℃下仅持续12 - 23天,在4℃下持续6个月。三种噬菌体分离株,即金黄色葡萄球菌噬菌体/CIRG/1、金黄色葡萄球菌噬菌体/CIRG/4和金黄色葡萄球菌噬菌体/CIRG/5,对超过80%的葡萄球菌分离株表现出裂解活性。本研究结果为使用裂解性噬菌体对引起山羊乳腺炎的多重耐药金黄色葡萄球菌进行治疗干预提供了依据。

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