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从感染 O157:H7 的鸽子粪便中分离、鉴定和比较基因组分析噬菌体 Ec_MI-02。

Isolation, Characterization, and Comparative Genomic Analysis of Bacteriophage Ec_MI-02 from Pigeon Feces Infecting O157:H7.

机构信息

Zayed Complex for Herbal Research and Traditional Medicine, Research and Innovation Center, Department of Health, Abu Dhabi 5674, United Arab Emirates.

Department of Biology, College of Science, United Arab Emirates University, Al Ain P.O. Box 15551, United Arab Emirates.

出版信息

Int J Mol Sci. 2023 May 30;24(11):9506. doi: 10.3390/ijms24119506.

Abstract

The most significant serotype of Shiga-toxigenic that causes foodborne illnesses is O157:H7. Elimination of O157:H7 during food processing and storage is a possible solution. Bacteriophages have a significant impact on bacterial populations in nature due to their ability to lyse their bacterial host. In the current study, a virulent bacteriophage, Ec_MI-02, was isolated from the feces of a wild pigeon in the United Arab Emirates (UAE) for potential future use as a bio-preservative or in phage therapy. Using a spot test and an efficiency of plating analysis, Ec_MI-02 was found to infect in addition to the propagation host, O157:H7 NCTC 12900, five different serotypes of O157:H7 (three clinical samples from infected patients, one from contaminated green salad, and one from contaminated ground beef). Based on morphology and genome analysis, Ec_MI-02 belongs to the genus Tequatrovirus under the order Caudovirales. The adsorption rate constant (K) of Ec_MI-02 was found to be 1.55 × 10 mL/min. The latent period was 50 min with a burst size of almost 10 plaque forming units (pfu)/host cell in the one-step growth curve when the phage Ec_MI-02 was cultivated using the propagation host O157:H7 NCTC 12900. Ec_MI-02 was found to be stable at a wide range of pH, temperature, and commonly used laboratory disinfectants. Its genome is 165,454 bp long with a GC content of 35.5% and encodes 266 protein coding genes. Ec_MI-02 has genes encoding for rI, rII, and rIII lysis inhibition proteins, which supports the observation of delayed lysis in the one-step growth curve. The current study provides additional evidence that wild birds could also be a good natural reservoir for bacteriophages that do not carry antibiotic resistance genes and could be good candidates for phage therapy. In addition, studying the genetic makeup of bacteriophages that infect human pathogens is crucial for ensuring their safe usage in the food industry.

摘要

引起食源性疾病的志贺毒素产毒型中最重要的血清型是 O157:H7。在食品加工和储存过程中消除 O157:H7 是一种可能的解决方案。噬菌体由于能够裂解其细菌宿主,因此对自然界中的细菌种群有重大影响。在当前的研究中,从阿拉伯联合酋长国(阿联酋)的一只野鸽粪便中分离出一种烈性噬菌体 Ec_MI-02,以备将来用作生物防腐剂或噬菌体治疗。通过斑点测试和效价分析,发现 Ec_MI-02 除了繁殖宿主 O157:H7 NCTC 12900 之外,还可以感染另外五种不同血清型的 O157:H7(来自感染患者的三种临床样本、一种来自污染的绿色沙拉和一种来自污染的碎牛肉)。根据形态和基因组分析,Ec_MI-02 属于尾病毒目 Caudovirales 下的 Tequatrovirus 属。Ec_MI-02 的吸附率常数(K)为 1.55×10 mL/min。在一步生长曲线中,噬菌体 Ec_MI-02 以繁殖宿主 O157:H7 NCTC 12900 为培养物时,潜伏期为 50 分钟,爆发量约为每个宿主细胞 10 个噬菌斑形成单位(pfu)。Ec_MI-02 在广泛的 pH、温度和常用实验室消毒剂下均稳定。它的基因组长 165,454 bp,GC 含量为 35.5%,编码 266 个蛋白编码基因。Ec_MI-02 具有编码 rI、rII 和 rIII 裂解抑制蛋白的基因,这支持了一步生长曲线中延迟裂解的观察结果。本研究进一步证明,野生鸟类也可能是不携带抗生素抗性基因的噬菌体的良好天然储库,并且可能是噬菌体治疗的良好候选者。此外,研究感染人类病原体的噬菌体的遗传组成对于确保它们在食品工业中的安全使用至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/111f/10253784/9d9b844662cc/ijms-24-09506-g001.jpg

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