Choi In Young, Park Do Hyeon, Chin Brayan A, Lee Cheonghoon, Lee Jinyoung, Park Mi-Kyung
School of Food Science and Biotechnology, and Food and Bio-Industry Research Institute, Kyungpook National University, Daegu 41566, Korea.
Materials Research and Education Center, Auburn University, Auburn, AL 36849, USA.
J Anim Sci Technol. 2020 Sep;62(5):668-681. doi: 10.5187/jast.2020.62.5.668. Epub 2020 Sep 30.
The purpose of this study was aimed to isolate a Typhimurium-specific phage (KFS-ST) from washing water in a poultry processing facility and to investigate the feasibility of the KFS-ST as a novel bio-receptor for the magnetoelastic (ME) biosensor method. KFS-ST against . Typhimurium was isolated, propagated, and purified using a CsCl-gradient ultracentrifugation. Morphological characteristics of KFS-ST were analyzed using transmission electron microscopy (TEM). Its specificity and efficiency of plating analysis were conducted against 39 foodborne pathogens. The temperature and pH stabilities of KFS-ST were investigated by the exposure of the phage to various temperatures (-70°C-70°C) and pHs (1-12) for 1 h. A one-step growth curve analysis was performed to determine the eclipse time, latent time and burst size of phage. The storage stability of KFS-ST was studied by exposing KFS-ST to various storage temperatures (-70°C, -20°C, 4°C, and 22°C) for 12 weeks. KFS-ST was isolated and purified with a high concentration of (11.47 ± 0.25) Log PFU/mL. It had an icosahedral head (56.91 ± 2.90 nm) and a non-contractile tail (225.49 ± 2.67 nm), which was classified into the family of in the order of . KFS-ST exhibited an excellent specificity against only Typhimurium and . Enteritidis, which are considered two of the most problematic strains in the meat and poultry. However, KFS-ST did not exhibit any specificity against six other and 27 non- strains. KFS-ST was stable at temperature of 4°C to 50°C and at pH of 4 to 12. The eclipse time, latent time, and burst size of KFS-ST were determined to be 10 min, 25 min and 26 PFU/ infected cell, respectively. KFS-ST was relatively stable during the 12-week storage period at all tested temperatures. Therefore, this study demonstrated the feasibility of KFS-ST as a novel bio-receptor for the detection of Typhimurium and Enteritidis in meat and poultry products using the ME biosensor method.
本研究旨在从家禽加工设施的冲洗水中分离出一种鼠伤寒沙门氏菌特异性噬菌体（KFS-ST），并研究KFS-ST作为磁弹性（ME）生物传感器方法新型生物受体的可行性。使用氯化铯梯度超速离心法分离、增殖并纯化了针对鼠伤寒沙门氏菌的KFS-ST。利用透射电子显微镜（TEM）分析了KFS-ST的形态特征。针对39种食源性病原体进行了其铺板分析的特异性和效率研究。通过将噬菌体暴露于不同温度（-70°C至70°C）和pH值（1至12）1小时，研究了KFS-ST的温度和pH稳定性。进行了一步生长曲线分析以确定噬菌体的隐蔽期、潜伏期和裂解量。通过将KFS-ST暴露于不同储存温度（-70°C、-20°C、4°C和22°C）12周，研究了KFS-ST的储存稳定性。KFS-ST以高浓度（11.47±0.25）Log PFU/mL被分离和纯化。它有一个二十面体头部（56.91±2.90nm）和一个非收缩性尾部（225.49±2.67nm），被归类为长尾噬菌体科长尾噬菌体属。KFS-ST仅对鼠伤寒沙门氏菌和肠炎沙门氏菌表现出优异的特异性，这两种菌被认为是肉类和家禽中最具问题的沙门氏菌菌株。然而，KFS-ST对其他六种沙门氏菌和27种非沙门氏菌菌株未表现出任何特异性。KFS-ST在4°C至50°C的温度和pH值为4至12时稳定。KFS-ST的隐蔽期、潜伏期和裂解量分别确定为10分钟、25分钟和26个PFU/感染细胞。在所有测试温度下，KFS-ST在12周储存期内相对稳定。因此，本研究证明了KFS-ST作为使用ME生物传感器方法检测肉类和家禽产品中鼠伤寒沙门氏菌和肠炎沙门氏菌的新型生物受体的可行性。
