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柱状黄杆菌特异性裂解噬菌体可使鲶鱼(塘鲺,林奈)免受柱状病侵害。

Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease.

作者信息

Prasad Yogendra, Kumar Dinesh, Sharma A K

机构信息

Aquatic Biotechnology and Fish Pathology Laboratory, Department of Animal Science, M.J.P. Rohilkhand University, Bareilly - 243 006, India.

出版信息

J Environ Biol. 2011 Mar;32(2):161-8.

Abstract

This investigation was aimed to find out appropriate strategy against antibiotic resistant bacterial fish pathogen, F. columnare. This pathogen was found persistently associated with fishes causing columnaris disease and ensuing mass mortality in hatchery and culture system of Sub - Himalayan region. Nine lytic F. columnare phages (FCP1 - FCP9) specific to its fifteen isolates were isolated from the water and bottom sediments of various geo-climatic regions of North India. The F. columnare phage FCP1 (made of hexagonal head and non contractile long tail belonging to family Podovariedae, a member of DNA virus) exhibited broader host range to lyse 9 out of 15 isolates of F. columnare. Therapeutic ability of FCP1 phage was assessed in C. batrachus inoculated intramuscularly (im) with virulent bacterial isolate FC8 and post inoculated (PI) with FCP1 phage (@ 10(8) : 10(6):: cfu : pfu) through intramuscular (im), immersion (bath) and oral (phage impregnated feed) treatment. Significant (p < 0.001) reduction (less than 10(-3) cfu ml(-1)) in host bacterium in the sera, gill, liver and kidney of challenged fishes was noted after 6 hr of phage treatment. Quantum of phage played a significant role in bringing down bacterial population as in the sera of dose 1 (@ 4.55 x 10(6) pfu ml(-1)) and dose 2 (@ 9.15 x 10(6) pfu ml(-1)) treated fishes mean log10 cfu value reduced by 3 logs (58.39%) and 5 logs (73.77%) at 96 hr, respectively. Phage treatment led to disappearance of gross symptoms, negative bacteriological test, detectable phage and 100% survival in experimentally infected C. batrachus. Result of this study provides evidence of profound lytic impact of FCP1 phage and represents its interesting therapeutic importance against antibiotic resistant F. columnare.

摘要

本研究旨在找出对抗抗生素耐药性鱼类病原菌柱状黄杆菌的合适策略。该病原菌一直与鱼类相关,在喜马拉雅地区的孵化场和养殖系统中引发柱状病并导致大量死亡。从印度北部不同地理气候区域的水体和底部沉积物中分离出了9种对其15个分离株具有裂解作用的柱状黄杆菌噬菌体(FCP1 - FCP9)。柱状黄杆菌噬菌体FCP1(由六边形头部和非收缩性长尾组成,属于短尾病毒科,是一种DNA病毒)对15个柱状黄杆菌分离株中的9个表现出更广泛的宿主范围并能将其裂解。通过肌肉注射(im)用强毒细菌分离株FC8接种牛蛙,并在接种后(PI)通过肌肉注射(im)、浸泡(浸浴)和口服(噬菌体浸渍饲料)处理给予FCP1噬菌体(@ 10(8) : 10(6):: cfu : pfu),评估FCP1噬菌体的治疗能力。噬菌体处理6小时后,在受攻击鱼类的血清、鳃、肝脏和肾脏中,宿主细菌显著(p < 0.001)减少(低于10(-3) cfu ml(-1))。噬菌体数量在降低细菌数量方面发挥了重要作用,如在剂量1(@ 4.55 x 10(6) pfu ml(-1))和剂量2(@ 9.15 x 10(6) pfu ml(-1))处理的鱼类血清中,在96小时时平均log10 cfu值分别降低了3个对数(58.39%)和5个对数(73.77%)。噬菌体处理导致实验感染的牛蛙中明显症状消失、细菌学检测呈阴性、可检测到噬菌体且存活率达100%。本研究结果提供了FCP1噬菌体具有深刻裂解作用的证据,并表明其在对抗抗生素耐药性柱状黄杆菌方面具有有趣的治疗重要性。

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