Maina Alice N, Mwaura Francis B, Wagacha John M, Jumba Miriam, Aziz Ramy K, Nour El-Din Hanzada T
Department of Biology, University of Nairobi, Nairobi, Kenya.
Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
J Basic Microbiol. 2023 May;63(5):481-488. doi: 10.1002/jobm.202200635. Epub 2023 Jan 20.
Bacteriophage therapy targeting the increasingly resistant Vibrio cholerae is highly needed. Hence, studying the phenotypic behavior of potential phages under different conditions is a prerequisite to delivering the phage in an active infective form. The objective of this study was to characterize phage VP4 (vB_vcM_Kuja), an environmental vibriophage isolated from River Kuja in Migori County, Kenya in 2015. The phenotypic characteristics of the phage were determined using a one-step growth curve, restriction digestion profile, pH, and temperature stability tests. The results revealed that the phage is stable through a wide range of temperatures (20-50°C) and maintains its plaque-forming ability at pH ranging from 6 to 12. The one-step growth curve showed a latent period falling between 40 and 60 min, while burst size ranged from 23 to 30 plaque-forming units/10 µl at the same host strain. The restriction digestion pattern using EcoRI, SalI, HindIII, and XhoI enzymes showed that HindIII could cut the phage genome. The phage DNA could not be restricted by the other three enzymes. The findings of this study can be used in future studies to determine phage-host interactions.
迫切需要针对日益耐药的霍乱弧菌的噬菌体疗法。因此,研究潜在噬菌体在不同条件下的表型行为是以活性感染形式递送噬菌体的先决条件。本研究的目的是对噬菌体VP4(vB_vcM_Kuja)进行特性分析,该噬菌体是2015年从肯尼亚米戈里县库贾河分离出的一种环境弧菌噬菌体。通过一步生长曲线、限制性消化图谱、pH和温度稳定性测试来确定该噬菌体的表型特征。结果显示,该噬菌体在很宽的温度范围(20-50°C)内都很稳定,并且在pH值为6至12的范围内保持其噬菌斑形成能力。一步生长曲线显示潜伏期在40至60分钟之间变化,而在相同宿主菌株中,裂解量范围为23至30个噬菌斑形成单位/10微升。使用EcoRI、SalI、HindIII和XhoI酶进行的限制性消化模式表明,HindIII可以切割噬菌体基因组。噬菌体DNA不能被其他三种酶限制。本研究的结果可用于未来研究以确定噬菌体-宿主相互作用。