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潜在噬菌体鸡尾酒制剂对从印度东南沿海对虾养殖环境中分离出的哈维氏弧菌及密切相关弧菌的功效。

Efficacy of potential phage cocktails against Vibrio harveyi and closely related Vibrio species isolated from shrimp aquaculture environment in the south east coast of India.

作者信息

Stalin Nattan, Srinivasan Pappu

机构信息

Molecular Biology Laboratory, Department of Bioinformatics, Alagappa University, Karaikudi 630 003, Tamilnadu, India.

Department of Animal Health and Management, Alagappa University, Karaikudi 630 003, Tamilnadu, India.

出版信息

Vet Microbiol. 2017 Aug;207:83-96. doi: 10.1016/j.vetmic.2017.06.006. Epub 2017 Jun 9.

Abstract

A diverse set of novel phages infecting the marine pathogenic Vibrio harveyi was isolated from shrimp aquaculture environments in the south east coast of India. Based on initial screening, three phages with a broad host range revealed that the growth inhibition of phage is relatively specific to V. harveyi. They were also able to infect V. alginolyticus and V. parahemolyticus that belonged to the Harveyi clade species from shrimp pond and sea coast environment samples. However, the impact of these phages on their host bacterium are well understood; a one-step growth curve experiment and transmission electron microscope (TEM) revealed three phages grouped under the Myoviridae (VHM1 and VHM2); Siphoviridae (VHS1) family. These phages were further molecular characterized with respect to phage genomic DNA isolates. The randomly amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) digestion with HindIII, and major structural proteins were distinguished by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) clearly indicated that all the phage isolates were different, even when they came from the same source, giving an insight into the diversity of phages. Evaluation of microcosm studies of Penaeus monodon larvae infected with V. harveyi (105 CFU mL-1) showed that larvae survival after 96 h in the presence of phage treatment at 109 PFU mL-1 was enhanced when compared with the control. The resolution in over survival highly recommended that this study provides the phage-based therapy which could be an innovative and eco-friendly solution against Vibrio disease in shrimp aquaculture and in the natural environment.

摘要

从印度东南沿海的对虾养殖环境中分离出了一组多样的新型噬菌体,它们可感染海洋致病性哈维氏弧菌。基于初步筛选,三种具有广泛宿主范围的噬菌体显示出噬菌体的生长抑制相对特异于哈维氏弧菌。它们还能够感染来自虾塘和海岸环境样本的属于哈维氏弧菌进化枝物种的溶藻弧菌和副溶血性弧菌。然而,这些噬菌体对其宿主细菌的影响尚不清楚;一步生长曲线实验和透射电子显微镜(TEM)显示三种噬菌体属于肌尾噬菌体科(VHM1和VHM2);长尾噬菌体科(VHS1)家族。这些噬菌体进一步通过噬菌体基因组DNA分离物进行分子特征分析。随机扩增多态性DNA(RAPD)、用HindIII进行的限制性片段长度多态性(RFLP)消化以及通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)区分主要结构蛋白,清楚地表明所有噬菌体分离物都是不同的,即使它们来自同一来源,这也深入了解了噬菌体的多样性。对感染哈维氏弧菌(105 CFU mL-1)的斑节对虾幼虫进行的微观研究评估表明,与对照相比,在109 PFU mL-1的噬菌体处理下96小时后幼虫的存活率有所提高。存活率的显著提高强烈表明,本研究提供了基于噬菌体的疗法,这可能是一种针对对虾养殖和自然环境中弧菌病的创新且环保的解决方案。

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