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猪霍乱沙门氏菌特异性噬菌体的筛选与鉴定

Selection and characterization of bacteriophages specific to Choleraesuis in swine.

作者信息

Sriprasong Pattaraporn, Imklin Napakhwan, Nasanit Rujikan

机构信息

Department of Biotechnology, Faculty of Engineering and Industrial Technology, Silpakorn University, Nakhon Pathom 73000, Thailand.

出版信息

Vet World. 2022 Dec;15(12):2856-2869. doi: 10.14202/vetworld.2022.2856-2869. Epub 2022 Dec 16.

Abstract

BACKGROUND AND AIM

Choleraesuis is the most common serotype that causes salmonellosis in swine. Recently, the use of bacteriophages as a potential biocontrol strategy has increased. Therefore, this study aimed to isolate and characterize bacteriophages specific to . Choleraesuis associated with swine infection and to evaluate the efficacy of individual phages and a phage cocktail against . Choleraesuis strains in simulated intestinal fluid (SIF).

MATERIALS AND METHODS

Three strains of . Choleraesuis isolated from pig intestines served as host strains for phage isolation. The other 10 serovars were also used for the phage host range test. The antibiotic susceptibility of the bacterial strains was investigated. Water samples from natural sources and drain liquid from slaughterhouses were collected for phage isolation. The isolated phages were characterized by determining the efficiency of plating against all Salmonella strains and the stability at a temperature range (4°C-65°C) and at low pH (2.5-4.0) in simulated gastric fluids (SGFs). Furthermore, morphology and genomic restriction analyses were performed for phage classification phages. Finally, . Choleraesuis reduction in the SIF by the selected individual phages and a phage cocktail was investigated.

RESULTS

The antibiotic susceptibility results revealed that most strains were sensitive to all tested drugs. Choleraesuis KPS615 was multidrug-resistant, showing resistance to three antibiotics. Nine phages were isolated. Most of them could infect four strains. Phages vB_SCh-RP5i3B and vB_SCh-RP61i4 showed high efficiency in infecting . Choleraesuis and Rissen. The phages were stable for 1 h at 4°C-45°C. However, their viability decreased when the temperature increased to 65°C. In addition, most phages remained viable at a low pH (pH 2.5-4.0) for 2 h in SGF. The efficiency of phage treatment against . Choleraesuis in SIF showed that individual phages and a phage cocktail with three phages effectively reduced . Choleraesuis in SIF. However, the phage cocktails were more effective than the individual phages.

CONCLUSION

These results suggest that the newly isolated phages could be promising biocontrol agents against S. Choleraesuis infection in pigs and could be orally administered. However, further studies should be conducted.

摘要

背景与目的

猪霍乱沙门氏菌是引起猪沙门氏菌病最常见的血清型。近年来,噬菌体作为一种潜在的生物防治策略的应用有所增加。因此,本研究旨在分离和鉴定与猪感染相关的猪霍乱沙门氏菌特异性噬菌体,并评估单个噬菌体和噬菌体鸡尾酒对模拟肠液(SIF)中猪霍乱沙门氏菌菌株的疗效。

材料与方法

从猪肠道分离的三株猪霍乱沙门氏菌用作噬菌体分离的宿主菌株。另外10个血清型也用于噬菌体宿主范围测试。研究了细菌菌株的抗生素敏感性。收集天然水源水样和屠宰场排水液用于噬菌体分离。通过测定对所有沙门氏菌菌株的平板接种效率以及在模拟胃液(SGF)中温度范围(4°C - 65°C)和低pH(2.5 - 4.0)下的稳定性来表征分离的噬菌体。此外,对噬菌体进行形态学和基因组限制性分析以进行噬菌体分类。最后,研究了所选单个噬菌体和噬菌体鸡尾酒对SIF中猪霍乱沙门氏菌的减少情况。

结果

抗生素敏感性结果显示,大多数菌株对所有测试药物敏感。猪霍乱沙门氏菌KPS615具有多重耐药性,对三种抗生素耐药。分离出九种噬菌体。其中大多数可以感染四种沙门氏菌菌株。噬菌体vB_SCh - RP5i3B和vB_SCh - RP61i4在感染猪霍乱沙门氏菌和里森沙门氏菌方面表现出高效率。噬菌体在4°C - 45°C下1小时内稳定。然而,当温度升至65°C时,它们的活力下降。此外,大多数噬菌体在SGF中低pH(pH 2.5 - 4.0)下2小时内仍保持活力。噬菌体处理对SIF中猪霍乱沙门氏菌的疗效表明,单个噬菌体和含有三种噬菌体的噬菌体鸡尾酒有效地减少了SIF中的猪霍乱沙门氏菌。然而,噬菌体鸡尾酒比单个噬菌体更有效。

结论

这些结果表明,新分离的噬菌体可能是用于防治猪感染猪霍乱沙门氏菌的有前景的生物防治剂,并且可以口服给药。然而,应进行进一步的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8a92/9880823/f9db7b0c4897/Vetworld-15-2856-g001.jpg

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