Department of Food Science and Human Nutrition, Iowa State University, Ames, IA, 50011, USA.
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, 5001 Campus Drive, College Park, MD, 20740, USA.
Anal Bioanal Chem. 2020 Mar;412(8):1741-1755. doi: 10.1007/s00216-020-02457-3. Epub 2020 Feb 11.
Previously, we demonstrated capture and concentration of Salmonella enterica subspecies enterica ser. Typhimurium using magnetic ionic liquids (MILs), followed by rapid isothermal detection of captured cells via recombinase polymerase amplification (RPA). Here, we report work intended to explore the broader potential of MILs as novel pre-analytical capture reagents in food safety and related applications. Specifically, we evaluated the capacity of the ([P][Ni(hfacac)]) ("Ni(II)") MIL to bind a wider range of human pathogens using a panel of Salmonella and Escherichia coli O157:H7 isolates, including a "deep rough" strain of S. Minnesota. We extended this exploration further to include other members of the family Enterobacteriaceae of food safety and clinical or agricultural significance. Both the Ni(II) MIL and the ([P][Dy(hfacac)]) ("Dy(III)") MIL were evaluated for their effects on cell viability and structure-function relationships behind observed antimicrobial activities of the Dy(III) MIL were determined. Next, we used flow imaging microscopy (FIM) of Ni(II) MIL dispersions made in model liquid media to examine the impact of increasing ionic complexity on MIL droplet properties as a first step towards understanding the impact of suspension medium properties on MIL dispersion behavior. Finally, we used FIM to examine interactions between the Ni(II) MIL and Serratia marcescens, providing insights into how the MIL may act to capture and concentrate Gram-negative bacteria in aqueous samples, including food suspensions. Together, our results provide further characterization of bacteria-MIL interactions and support the broader utility of the Ni(II) MIL as a cell-friendly capture reagent for sample preparation prior to cultural or molecular analyses. Graphical abstract.
先前,我们利用磁性离子液体 (MILs) 捕获并浓缩沙门氏菌亚种肠炎血清型 Typhimurium,随后通过重组聚合酶扩增 (RPA) 快速等温检测捕获的细胞。在此,我们报告了旨在探索 MILs 在食品安全和相关应用中作为新型分析前捕获试剂的更广泛潜力的工作。具体而言,我们使用沙门氏菌和大肠杆菌 O157:H7 分离株的面板评估了 ([P][Ni(hfacac)])(“Ni(II)”) MIL 结合更广泛范围的人类病原体的能力,包括 S. Minnesota 的“深粗糙”菌株。我们进一步扩展了这项探索,包括食品安全性和临床或农业意义上的其他肠杆菌科成员。我们评估了 Ni(II) MIL 和 ([P][Dy(hfacac)])(“Dy(III)”) MIL 对细胞活力的影响,并确定了 Dy(III) MIL 观察到的抗菌活性背后的结构-功能关系。接下来,我们使用模型液体介质中 Ni(II) MIL 分散体的流动成像显微镜 (FIM) 检查了增加离子复杂性对 MIL 液滴性质的影响,作为了解悬浮介质性质对 MIL 分散行为影响的第一步。最后,我们使用 FIM 检查了 Ni(II) MIL 与粘质沙雷氏菌之间的相互作用,深入了解了 MIL 如何在水溶液样品(包括食品悬浮液)中捕获和浓缩革兰氏阴性细菌。总的来说,我们的结果进一步说明了细菌-MIL 相互作用的特性,并支持 Ni(II) MIL 作为用于文化或分子分析之前的样品制备的友好型细胞捕获试剂的更广泛用途。