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感染耐碳青霉烯类致病性腹泻病原菌和尿路病原菌的新型裂解性噬菌体的分离与鉴定

Isolation and Characterization of Novel Lytic Bacteriophages Infecting Carbapenem-Resistant Pathogenic Diarrheagenic and Uropathogenic .

作者信息

Ssekatawa Kenneth, Ntulume Ibrahim, Byarugaba Denis Karuhize, Michniewski Slawomir, Jameson Eleanor, Wampande Eddie M, Nakavuma Jesca

机构信息

Department of Science Technical and Vocational Education, Makerere University, Kampala, Uganda.

Africa Center of Excellence in Materials, Product Development and Nanotechnology (MAPRONANO ACE), Makerere University, Kampala, Uganda.

出版信息

Infect Drug Resist. 2024 Aug 6;17:3367-3384. doi: 10.2147/IDR.S466101. eCollection 2024.

Abstract

BACKGROUND

The evolution of antimicrobial resistance has dramatically reduced the efficacy of the first-choice and last-resort antibiotics used to treat infections. Thus, searching for novel therapeutics to treat and control the emergence of antibiotic resistance is urgent. Therefore, this study aimed to illustrate the lytic effect of phages against carbapenem-resistant pathogenic .

METHODS

Phages were isolated from hospital effluents by the enrichment assay. This was followed by the evaluation of the host range of the phages by the spot assay. The time taken by phages to bind to the host bacterial cells was determined by the adsorption assay. The phage latent period and burst size were determined using a one-step growth experiment. Phage morphology was determined by the Transmission Electron Microscopy. Molecular characterization of phages was done by whole genome sequencing.

RESULTS

Two phages named UGKSEcP1 and UGKSEcP2 were isolated from hospital effluents. The phages were professionally lytic with a broad host range. The two phages recorded an average adsorption time of 11.25 minutes, an adsorption rate of 99.3%, a latency period of 20 minutes, and a burst size of approximately 528 phages/infected cell. Phages UGKSEcP1 and UGKSEcP2 had genome lengths of 167433bp, and 167221bp with 277 and 276 predicted genes, respectively, and no undesirable genes were detected. Phylogenetic analysis revealed the two phages belonged genus Tequatrovirus. TEM micrograph showed that the two phages had a similar morphotype with icosahedral heads and contractile tails; thus, classified as members of the Myoviridae phage family.

CONCLUSION

The findings demonstrate that the study isolated two novel professionally lytic phages with a broad host range and thus, are candidates for phage-mediated biocontrol.

摘要

背景

抗菌药物耐药性的演变极大地降低了用于治疗感染的一线和最后手段抗生素的疗效。因此,寻找治疗和控制抗生素耐药性出现的新型疗法迫在眉睫。因此,本研究旨在阐明噬菌体对耐碳青霉烯类病原菌的裂解作用。

方法

通过富集试验从医院污水中分离噬菌体。随后通过斑点试验评估噬菌体的宿主范围。通过吸附试验确定噬菌体与宿主细菌细胞结合所需的时间。使用一步生长实验确定噬菌体的潜伏期和爆发量。通过透射电子显微镜确定噬菌体形态。通过全基因组测序对噬菌体进行分子特征分析。

结果

从医院污水中分离出两种噬菌体,命名为UGKSEcP1和UGKSEcP2。这些噬菌体具有专业的裂解能力,宿主范围广泛。这两种噬菌体的平均吸附时间为11.25分钟,吸附率为99.3%,潜伏期为20分钟,爆发量约为528个噬菌体/感染细胞。噬菌体UGKSEcP1和UGKSEcP2的基因组长度分别为167433bp和167221bp,分别有277个和276个预测基因,未检测到不良基因。系统发育分析表明这两种噬菌体属于Tequatrovirus属。透射电子显微镜照片显示这两种噬菌体具有相似的形态类型,头部为二十面体且尾部可收缩;因此,被归类为肌尾噬菌体科的成员。

结论

研究结果表明,本研究分离出两种新型的具有广泛宿主范围的专业裂解噬菌体,因此是噬菌体介导生物防治的候选者。

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