Rahimzadeh Golnar, Gill Pooria, Rezai Mohammad Sadegh
Pediatric Infectious Diseases Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran.
Iran J Basic Med Sci. 2019 Mar;22(3):290-295. doi: 10.22038/ijbms.2019.31226.7521.
During the last years with increasing resistant bacteria to the most antibiotics bacteriophages are suggested as appropriate treatment option. To investigate lytic activity of bacteriophages there are indirect microbial procedures and direct methods. The present study to complement microbial procedures and investigate ultra-structural characteristics of infection bacterium-phage use atomic force microscopy technique.
The bacteriophages were isolated from sewage at the Tertiary Pediatric Hospital. Bacteriophages (10×10 PFU/ml) were diluted and were mixed with 100 μl of methicillin resistant (MRSA) ATCC 33591 (1.5×10 CFU/ml). The tubes were incubated for 20 min at 37 C, at intervals 10 min, 10 μl samples were removed and directly were investigated MRSA ATCC morphology, roughness parameter, 3D topography, cell height, and fast Fourier transform (FFT) by atomic force microscopy (AFM) technique. Concurrently turbidity assay were performed.
Concentration of MRSA ATCC No. 33591 strain after 10 min in phage-treated MRSA S3 (1.5×10 CFU/ml), S4 (1.5×10 CFU/ml), S5 (1.5×10 CFU/ml), S6 (1.5×10 CFU/ml) decreased 2-log, 3-log, 4-log, and 5-log respectively. The results AFM micrographs shown the most changes in bacterial morphology and 3D topography, destruction of cell wall, decrease of cell height, and loss of their shape after 10 min at phage-treated MRSA S3 (1.5×10 CFU/ml), S4 (1.5×10 CFU/ml), S5 (1.5×10 CFU/ml), S6 (1.5×10 CFU/ml) respectively .
In this study MRSA ATCC ultra-structural changes in phage-treated MRSA ATCC groups directly were detected using AFM technique.
在过去几年中，随着对大多数抗生素耐药的细菌不断增加，噬菌体被认为是一种合适的治疗选择。为了研究噬菌体的裂解活性，有间接微生物程序和直接方法。本研究旨在补充微生物程序，并使用原子力显微镜技术研究感染细菌 - 噬菌体的超微结构特征。
噬菌体从三级儿童医院的污水中分离出来。将噬菌体（10×10 PFU/ml）稀释并与100μl耐甲氧西林金黄色葡萄球菌（MRSA）ATCC 33591（1.5×10 CFU/ml）混合。将试管在37℃下孵育20分钟，每隔10分钟取出10μl样品，并通过原子力显微镜（AFM）技术直接研究MRSA ATCC的形态、粗糙度参数、三维形貌、细胞高度和快速傅里叶变换（FFT）。同时进行浊度测定。
在噬菌体处理的MRSA S3（1.5×10 CFU/ml）、S4（1.5×10 CFU/ml）、S5（1.5×10 CFU/ml）、S6（1.5×10 CFU/ml）中，10分钟后MRSA ATCC 33591菌株的浓度分别下降了2个对数、3个对数、4个对数和5个对数。原子力显微镜图像结果显示，在噬菌体处理的MRSA S3（1.5×10 CFU/ml）、S4（1.5×10 CFU/ml）、S5（1.5×10 CFU/ml）、S6（1.5×10 CFU/ml）中，10分钟后细菌形态和三维形貌变化最大，细胞壁破坏，细胞高度降低，形状丧失。
在本研究中，使用原子力显微镜技术直接检测了噬菌体处理的MRSA ATCC组中MRSA ATCC的超微结构变化。
