Characterization of Pseudomonas lytic phages and their application as a cocktail with antibiotics in controlling Pseudomonas aeruginosa
Characterization of Pseudomonas lytic phages and their application as a cocktail with antibiotics in controlling Pseudomonas aeruginosa
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DOI：
10.1016/j.jbiosc.2020.02.001
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发表时间：
2020-06-01
影响因子：
2.8
通讯作者：
Tanji, Yasunori
中科院分区：
文献类型：
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作者：
Ong, Soo Peng;Azam, Aa Haeruman;Tanji, Yasunori
铜绿假单胞菌是一种条件致病菌，可引起免疫功能低下和慢性囊性纤维化（CF）患者的院内疾病。我们表征了两种新分离的具有不同宿主谱的铜绿假单胞菌，phi PA 01和phi PA 02，并检查了它们作为抗生素对铜绿假单胞菌的鸡尾酒的效果，以表明将噬菌体鸡尾酒和抗生素组合治疗假单胞菌感染的可能性。噬菌体phi PA 01（66，220 bp）和phi PA 02（279，095 bp）分别属于普布那病毒属和菲克斯病毒属。在它们的基因组中未发现毒力或溶原性相关基因，因此它们用于噬菌体治疗是潜在安全的。我们产生了各自的噬菌体抗性菌株，以研究两种细菌之间的交叉抗性。在phi PA 01抗性菌株中观察到对phi PA 02的轻微交叉抗性，而phi PA 02抗性菌株仍然对phi PA 01敏感。在第5轮出现了一株与phi PA 02交叉耐药的phi PA 01耐药菌株（R5-PA 01 R），该菌株在核心脂多糖（LPS）合成的关键酶磷酸葡萄糖变位酶（algC）上发生了移码突变。algC基因敲除对两种酶都有抗性。Δ algC的互补恢复了LPS的感染性，表明LPS作为宿主受体。与单一噬菌体（8 e9 h）相比，噬菌体混合物抑制铜绿假单胞菌生长更长（20 h）小时，进一步表明它们用作噬菌体混合物的潜力。此外，应用含有环丙沙星（0.25 μ g/ ml）和美罗培南（2 μ g/ml）的噬菌体混合物，能够抑制铜绿假单胞菌的生长长达96小时。我们的研究结果显示了phi PA 01和phi PA 02作为噬菌体混合物与抗生素一起用于治疗铜绿假单胞菌的潜在应用。(C)2020年，生物技术学会，日本。All rights reserved.
Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial disease among immunocompromised and chronic cystic fibrosis (CF) patients. We characterized two newly isolated Pseudomonas phages, phi PA01 and phi PA02, with different host spectra, and examined their effect as a cocktail with antibiotics against P. aeruginosa, to indicate the possibility of combining a phage cocktail and antibiotics in treating pseudomonal infection. Phages phi PA01 (66,220 bp) and phi PA02 (279,095 bp) belong to the genus Pbunalikevirus and Phikzlikevirus, respectively. No virulence or lysogenic associated gene was found in their genomes, thus they are potentially safe for phage therapy. We generated respective phage-resistant strains to investigate cross-resistance between two phages. Slight cross-resistance to phi PA02 in phi PA01-resistant strain was observed, while phi PA02-resistant strain remained susceptible to phi PA01. A phi PA01 resistant strain that was cross-resistant to phi PA02 appeared in round 5 (R5-PA01R), revealed frameshift mutation in phosphoglucomutase (algC), which is important for the synthesis of core lipopolysaccharide (LPS). Knockout of algC was resistant to both phages. Complementation of Delta algC restored phages' infectivity, suggesting that LPS as host receptor. Phage cocktail suppressed the growth of P. aeruginosa for longer (20 h) hour compared with single phage (8e9 h), further suggesting their potential to be used as a phage cocktail. Furthermore, application of the phage cocktail with ciprofloxacin (0.25 mu g/ ml) and meropenem (2 mu g/ml), managed to suppress the growth of P. aeruginosa up to 96 h. Our results show the potential application of phi PA01 and phi PA02 as phage cocktail together with antibiotics for treatment of P. aeruginosa. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.