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从亚的斯亚贝巴的各种来源中分离和鉴定针对抗微生物耐药性腹泻性大肠杆菌菌株的裂解噬菌体,并评估其治疗潜力。

Isolation and characterization of lytic bacteriophages from various sources in Addis Ababa against antimicrobial-resistant diarrheagenic Escherichia coli strains and evaluation of their therapeutic potential.

机构信息

Institute of Biotechnology, Addis Ababa University, P.O.Box 1176, Addis Ababa, Ethiopia.

Department of Biotechnology, Woldia University, P.O. Box 400, Woldia, Ethiopia.

出版信息

BMC Infect Dis. 2024 Mar 14;24(1):310. doi: 10.1186/s12879-024-09152-z.

Abstract

BACKGROUND

Escherichia coli is a common fecal coliform, facultative aerobic, gram-negative bacterium. Pathogenic strains of such microbes have evolved to cause diarrhea, urinary tract infections, and septicemias. The emergence of antibiotic resistance urged the identification of an alternative strategy. The use of lytic bacteriophages against the control of pathogenic E. coli in clinics and different environmental setups (waste and drink water management) has become an alternative therapy to antibiotic therapy. Thus, this study aimed to isolate and characterize lytic bacteriophage from various sources in Addis Ababa, tested them against antimicrobial-resistant diarrheagenic E. coli strains and evaluated their therapeutic potential under in vitro conditions.

METHODS

A total of 14 samples were processed against six different diarrheagenic E. coli strains. The conventional culture and plaque analysis agar overlay method was used to recover lytic bacteriophage isolates. The phage isolates were characterized to determine their lytic effect, growth characteristics, host range activity, and stability under different temperature and pH conditions. Phage isolates were identified by scanning electron microscope (SEM), and molecular techniques (PCR).

RESULTS

In total, 17 phages were recovered from 84 tested plates. Of the 17 phage isolates, 11 (65%) were Myoviridae-like phages, and 6 (35%) phage isolates were Podoviridae and Siphoviridae by morphology and PCR identification. Based on the host range test, growth characteristics, and stability test 7 potent phages were selected. These phages demonstrated better growth characteristics, including short latent periods, highest burst sizes, and wider host ranges, as well as thermal stability and the ability to survive in a wide range of pH levels.

CONCLUSIONS

The promising effect of the phages isolated in this study against AMR pathogenic E. coli has raised the possibility of their use in the future treatment of E. coli infections.

摘要

背景

大肠杆菌是一种常见的粪便革兰氏阴性需氧杆菌。此类微生物的致病性菌株已经进化到能够引起腹泻、尿路感染和败血症。抗生素耐药性的出现促使人们寻找替代策略。在临床和不同环境设置(废物和饮用水管理)中,使用裂解噬菌体来控制致病性大肠杆菌已成为抗生素治疗的替代疗法。因此,本研究旨在从亚的斯亚贝巴的各种来源中分离和鉴定裂解噬菌体,测试它们对抗抗菌药物耐药性腹泻性大肠杆菌菌株的作用,并在体外条件下评估它们的治疗潜力。

方法

用常规培养和噬菌斑分析琼脂覆盖法从 14 个样本中回收裂解噬菌体。对噬菌体分离物进行鉴定,以确定其裂解作用、生长特性、宿主范围活性以及在不同温度和 pH 条件下的稳定性。通过扫描电子显微镜(SEM)和分子技术(PCR)对噬菌体分离物进行鉴定。

结果

从 84 个测试平板中总共回收了 17 个噬菌体。在 17 个噬菌体分离物中,11 个(65%)是肌尾噬菌体样噬菌体,6 个(35%)噬菌体分离物通过形态学和 PCR 鉴定为 Podoviridae 和 Siphoviridae 噬菌体。根据宿主范围试验、生长特性和稳定性试验,选择了 7 种有效的噬菌体。这些噬菌体表现出更好的生长特性,包括较短的潜伏期、最高的爆发量和更广泛的宿主范围,以及热稳定性和在广泛 pH 水平下的生存能力。

结论

本研究中分离的噬菌体对 AMR 致病性大肠杆菌的有前景的效果,提高了它们在未来治疗大肠杆菌感染中的应用可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff2a/10938718/4c5f0c706cb2/12879_2024_9152_Fig1_HTML.jpg

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