Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen Key Laboratory of Marine Bioresource and Eco-environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, 518055, China.
Probiotics Antimicrob Proteins. 2019 Jun;11(2):667-675. doi: 10.1007/s12602-018-9446-3.
The antibiotic-resistant viable but non-culturable (VBNC) pathogenic bacteria are considered as a new threat to public health. Antimicrobial peptides (AMPs), possessing bactericidal effects in a rapid membrane attacking mode, are supposed to be effective against bacteria entering the VBNC state. In the current study, the activity of grouper AMP piscidin killing the VBNC state cells of pathogenic bacteria Escherichia coli O157, Staphylococcus aureus, and Vibrio parahaemolyticus OS4 was studied. After entering the VBNC state, cells of E. coli O157, S. aureus, and V. parahaemolyticus OS4 developed resistance to the antibiotics Ampicillin and Kanamycin. Rather than truncated form of Malabar grouper piscidin 1 (EmPis-1S), full-length Malabar grouper piscidin 1 (EmPis-1L) showed strong activity to kill the above VBNC bacteria. The VBNC state cells (1 × 10 CFU/mL) of the three species of bacteria could be totally lysed by 10 μmol/L of EmPis-1L in 1 h. The VBNC state cells of S. aureus were most susceptible to EmPis-1L, which killed the cells by 100% in 30 min at the low concentration of 2.0 μmol/L. In addition, EmPis-1L at the concentration of no more than 10 μmol/L showed no observed toxicity to human lung carcinoma epithelial cells (A549) and mouse neuroblastoma cells (N2a). Accordingly, EmPis-1L could be a promisingly safe and efficient agent for eliminating the traditional antibiotic-resistant VBNC state cells of pathogenic bacteria, E. coli, S. aureus, and V. parahaemolyticus.
抗生素耐药的活但非可培养（VBNC）致病性细菌被认为是对公共卫生的新威胁。抗菌肽（AMPs）以快速的膜攻击模式具有杀菌作用，被认为对进入 VBNC 状态的细菌有效。在本研究中，研究了石斑鱼 AMP 鱼精蛋白杀死致病性细菌大肠杆菌 O157、金黄色葡萄球菌和副溶血性弧菌 OS4 的 VBNC 状态细胞的活性。进入 VBNC 状态后，大肠杆菌 O157、金黄色葡萄球菌和副溶血性弧菌 OS4 的细胞对氨苄西林和卡那霉素等抗生素产生耐药性。全长石斑鱼鱼精蛋白 1（EmPis-1L）而非截短形式的 Malabar 石斑鱼鱼精蛋白 1（EmPis-1S）对杀死上述 VBNC 细菌表现出很强的活性。三种细菌的 1×10 CFU/mL 的 VBNC 状态细胞可以在 1 小时内完全被 10 μmol/L 的 EmPis-1L 裂解。金黄色葡萄球菌的 VBNC 状态细胞对 EmPis-1L 最敏感，在 2.0 μmol/L 的低浓度下 30 分钟内即可杀死 100%的细胞。此外，浓度不超过 10 μmol/L 的 EmPis-1L 对人肺癌上皮细胞（A549）和小鼠神经母细胞瘤细胞（N2a）没有观察到毒性。因此，EmPis-1L 可能是一种有前途的安全有效的清除传统抗生素耐药的致病性细菌（大肠杆菌、金黄色葡萄球菌和副溶血性弧菌）VBNC 状态细胞的药物。
