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Survival and Transfer of Murine Norovirus within a Hydroponic System during Kale and Mustard Microgreen Harvesting

机译:羽衣甘蓝和芥菜微绿收获期水培系统中鼠诺如病毒的存活和转移

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Hydroponically grown microgreens are gaining in popularity, but there is a lack of information pertaining to their microbiological safety. The potential risks associated with virus contamination of crops within a hydroponic system have not been studied to date. Here a human norovirus (huNoV) surrogate (murine norovirus [MNV]) was evaluated for its ability to become internalized from roots to edible tissues of microgreens. Subsequently, virus survival in recirculated water without adequate disinfection was assessed. Kale and mustard seeds were grown on hydroponic pads (for 7 days with harvest at days 8 to 12), edible tissues (10 g) were cut 1 cm above the pads, and corresponding pieces (4 cm by 4 cm) of pads containing only roots were collected separately. Samples were collected from a newly contaminated system (recirculated water inoculated with ~3 log PFU/ml MNV on day 8) and from a previously contaminated system. (A contaminated system without adequate disinfection or further inoculation was used for production of another set of microgreens.) Viral titers and RNA copies were quantified by plaque assay and real-time reverse transcription (RT)-PCR. The behaviors of MNV in kale and mustard microgreens were similar ( P > 0.05). MNV was detected in edible tissues and roots after 2 h postinoculation, and the levels were generally stable during the first 12 h. Relatively low levels (~2.5 to ~1.5 log PFU/sample of both edible tissues and roots) of infectious viruses were found with a decreasing trend over time from harvest days 8 to 12. However, the levels of viral RNA present were higher and consistently stable (~4.0 to ~5.5 log copies/sample). Recirculated water maintained relatively high levels of infectious MNV over the period of harvest, from 3.54 to 2.73 log PFU/ml. Importantly, cross-contamination occurred easily; MNV remained infectious in previously contaminated hydroponic systems for up to 12 days (2.26 to 1.00 PFU/ml), and MNV was detected in both edible tissues and roots. Here we see that viruses can be recirculated in water, even after an initial contamination event is removed, taken up through the roots of microgreens, and transferred to edible tissues. The ease of product contamination shown here reinforces the need for proper sanitation.
机译:水培法种植的绿色植物越来越受欢迎,但是缺乏有关其微生物安全性的信息。迄今为止,尚未研究过与水培系统内农作物病毒污染有关的潜在风险。在此,评估了人类诺如病毒(huNoV)替代品(鼠诺如病毒[MNV])其从根到微绿的可食用组织的内在化能力。随后,评估了未经充分消毒的循环水中病毒的存活率。羽衣甘蓝和芥菜籽在水培垫上生长(在收获后的8到12天共收获7天),将可食用的组织(10 g)从垫上切开1厘米,相应的垫块(4厘米乘4厘米)仅包含分别收集根。从新污染的系统(第8天接种约3 log PFU / ml MNV的循环水)中收集样品。 (使用未经充分消毒或进一步接种的受污染系统来生产另一套微绿。)通过噬斑测定和实时逆转录(RT)-PCR对病毒滴度和RNA拷贝进行定量。芥蓝和芥末微果肉中MNV的行为相似(P> 0.05)。接种后2 h,在食用组织和根中检测到MNV,在最初的12 h中,该水平通常稳定。从收获的第8天到第12天,发现相对较低的感染性病毒水平(〜2.5至〜1.5 log PFU /可食用组织和根的样本)随时间呈下降趋势。但是,存在的病毒RNA水平较高且一致稳定(约4.0至约5.5日志副本/样本)。在收获期间,循环水保持了较高的传染性MNV水平,从3.54到2.73 log PFU / ml。重要的是,容易发生交叉污染。 MNV在先前受污染的水培系统中保持感染长达12天(2.26至1.00 PFU / ml),并且在食用组织和根部均检测到MNV。在这里,我们看到,即使去除了最初的污染事件,病毒也可以在水中再循环,通过微型果树的根部吸收并转移到可食用的组织中。此处显示的产品污染容易程度加剧了对适当卫生设施的需求。

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