Isolation and Identification of Lytic Phages Infecting Proteus mirabilis from Canine-derived and Their Effects on Biofilm |
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| Authors: | SONG Jun XU Ruoyang JIN Yuqi CHEN Li WANG Yuhui RUAN Hongri ZHENG Jiasan WU Rui |
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| Institution: | College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China |
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| Abstract: | The present study aimed to isolate phages that could lyse Proteus mirabilis from sewage in Daqing, Heilongjiang, and analyze their biological characteristics and effects on biofilm and provide the basic data for phage therapy as well as prevention. Phages were isolated and purified from sewage by double plate culture method, and their plaque morphology were observed. Morphological characteristics of purified phages were observed by transmission electron microscopy. The optimal multiplicity of infection (MOI), one-step growth curve, thermal stability, pH, nucleic acids analysis and the inhibition and removal effect of biofilm were determined. Three lytic of bacteriophages of Proteus mirabilis were successfully isolated and purified, and named as vB_PmM_S, vB_PmM_W and vB_PmM_X. The phage plaques were circular, clear and transparent with smooth edge. The electron microscope observation showed that the phage particles, those head was twenty-side, head length was about (30±3) nm. The optical MOI was 0.001, the incubation period was 20 min, the burst period was 35 min, and the burst size was 70 PFU·cell-1. The phages could withstand the temperature range from 10 to 60 ℃ and keep stable titer under pH 5.0-8.0. The phage vB_PmM_S can be stored at 4 ℃ in SM solution for a short time, and stored at -80 ℃ in glycerin (30%) or DMSO (5%) for a long time. The mixture of three bacteriophages can inhibit the biofilm formation and remove the biofilm. The results laid the foundation for the treatment of Proteus mirabilis and urinary tract infections. |
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| Keywords: | canine Proteus mirabilis lytic bacteriophage biological characteristics biofilm |
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