Thapa Kanchan, Julianingsih Dita, Tung Chuan-Wei, Phan Anna, Hashmi Muhammad Abrar, Bleich Kayla, Biswas Debabrata
Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.
Biological Sciences Program, Molecular and Cellular Biology, University of Maryland, College Park, MD 20742, USA.
Foods. 2024 Aug 29;13(17):2746. doi: 10.3390/foods13172746.
Enterohemorrhagic (EHEC) outbreaks have been frequently linked to the consumption of produce. Furthermore, produce grown on organic farms possess a higher risk, as the farmers avoid antibiotics and chemicals. This study sets out to evaluate the effectiveness of advanced postharvest disinfection processes using berry pomace extracts (BPEs) in reducing EHEC load in two common leafy greens, spinach and lettuce. Spinach and lettuce were inoculated with 5 log CFU/leaf EHEC EDL-933 and then treated with three different concentrations of BPE (1, 1.5, and 2 gallic acid equivalent, GAE mg/mL) for increasing periods of time. After the wash, the bacteria were quantified. Changes in the relative expression of virulence genes and the genes involved in cell division and replication and response against stress/antibiotics were studied. We observed a significant reduction in EHEC EDL933, ranging from 0.5 to 1.6 log CFU/spinach leaf ( < 0.05) washed with BPE water. A similar trend of reduction, ranging from 0.3 to 1.3 log CFU/mL, was observed in pre-inoculated lettuce washed with BPE water. We also quantified the remaining bacterial population in the residual treatment solutions and found the survived bacterial cells (3 log CFU/mL) were low despite repeated washing with the same solution. In addition, we evaluated the phenolic concentration in leftover BPE, which did not change significantly, even after multiple uses. Alterations in gene expression levels were observed, with downregulation ranging from 1 to 3 log folds in the genes responsible for the adhesion and virulence of EHEC EDL933 and significant upregulation of genes responsible for survival against stress. All other genes were upregulated, ranging from 2 to 7 log folds, with a dose-dependent decrease in expression. This finding shows the potential of BPE to be used for sanitation of fresh produce as a natural and sustainable approach.
出血性大肠杆菌(EHEC)疫情频繁与农产品消费相关联。此外,有机农场种植的农产品风险更高,因为农民避免使用抗生素和化学品。本研究旨在评估使用浆果渣提取物(BPEs)的先进采后消毒工艺在降低两种常见绿叶蔬菜(菠菜和生菜)中EHEC负荷方面的有效性。菠菜和生菜接种约5 log CFU/叶的EHEC EDL-933,然后用三种不同浓度的BPE(1、1.5和2没食子酸当量,GAE mg/mL)处理不同时间。清洗后,对细菌进行定量。研究了毒力基因以及参与细胞分裂、复制和应激/抗生素反应的基因的相对表达变化。我们观察到,用BPE水清洗的菠菜叶中,EHEC EDL933显著减少,减少范围为0.5至1.6 log CFU/菠菜叶(P<0.05)。在用BPE水清洗的预接种生菜中也观察到类似的减少趋势,减少范围为0.3至1.3 log CFU/mL。我们还对残留处理溶液中的剩余细菌种群进行了定量,发现尽管用相同溶液反复清洗,存活的细菌细胞(约3 log CFU/mL)数量较低。此外,我们评估了剩余BPE中的酚类浓度,即使多次使用后也没有显著变化。观察到基因表达水平发生改变,负责EHEC EDL933黏附和毒力的基因下调1至3 log倍,负责应激存活的基因显著上调。所有其他基因上调,范围为2至7 log倍,表达呈剂量依赖性下降。这一发现表明BPE作为一种天然且可持续的方法,有潜力用于新鲜农产品的卫生处理。