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Biofunctionalization of Cork with Moringa oleifera Seeds and Use of PMA Staining and qPCR to Detect Viability of Escherichia coli
Water ( IF 3.0 ) Pub Date : 2021-10-02 , DOI: 10.3390/w13192731 Nury Infante , Refugio Rodríguez , Yaneth Bartolo , Olga Sánchez , Isabel Sanz , Lizeth Bermeo , Jordi Morató
Water ( IF 3.0 ) Pub Date : 2021-10-02 , DOI: 10.3390/w13192731 Nury Infante , Refugio Rodríguez , Yaneth Bartolo , Olga Sánchez , Isabel Sanz , Lizeth Bermeo , Jordi Morató
Cork matrices biofunctionalized with Moringa oleifera seed extracts (MoSe) have potential for use as a biofilter with antibacterial properties to reduce waterborne pathogens. The aim of this study was to evaluate the effect of cork biofunctionalized with active antimicrobial compounds of MoSe (f-cork) on the inhibition of Escherichia coli (InhEc). The LacZ gene from a strain of E. coli was used as the target sequence using viability quantification Polymerase Chain Reaction (qPCR) and differentiation of viable and dead bacteria through selective cell viability PMA staining. To perform this, a 27−4 fractional factorial design and a biofiltration system were used to evaluate the effect of the active protein in MoSe immobilized in granulated cork on InhEc. We found that the potential for antimicrobial activity increased with f-cork for an effective maximal bacterial reduction (99.99%; p < 0.05). The effect of f-cork functionalized with MoSe on E. coli viability was of 0.024% and 0.005% for the cells exposed to PMA, respectively, being the relevant conditions in treatment 2: (0 L/min) without aeration, (5%) MoSe and (5 mm) cork particle. In conclusion, the f-cork functionalized with MoSe presented biosorbent and antibacterial properties that effectively reduced the E. coli growth.
中文翻译:
软木塞与辣木种子的生物功能化和使用 PMA 染色和 qPCR 检测大肠杆菌的活力
用辣木种子提取物 ( Mo Se)生物功能化的软木基质具有用作生物过滤器的潜力,具有抗菌特性,可减少水生病原体。本研究的目的是评估用Mo Se ( f- cork) 的活性抗菌化合物生物功能化的软木对抑制大肠杆菌(Inh Ec ) 的影响。使用活力定量聚合酶链反应 (qPCR) 和通过选择性细胞活力 PMA 染色区分活细菌和死细菌,将来自大肠杆菌菌株的LacZ基因用作目标序列。为此,一个 2 7−4部分因子设计和生物过滤系统用于评估固定在颗粒状软木中的MoSe中的活性蛋白对InhEc 的影响。我们发现抗微生物活性的潜力随着f- cork 的增加而增加,从而有效地最大程度地减少细菌 (99.99%;p < 0.05)。对于暴露于 PMA 的细胞,用Mo Se功能化的f- cork对大肠杆菌生存力的影响分别为 0.024% 和 0.005%,这是处理 2 中的相关条件:(0 L/min)无通气,(5 %) Mo Se 和 (5 mm) 软木颗粒。总之,用Mo功能化的f- corkSe 具有生物吸附和抗菌特性,可有效减少大肠杆菌的生长。
更新日期:2021-10-02
中文翻译:
软木塞与辣木种子的生物功能化和使用 PMA 染色和 qPCR 检测大肠杆菌的活力
用辣木种子提取物 ( Mo Se)生物功能化的软木基质具有用作生物过滤器的潜力,具有抗菌特性,可减少水生病原体。本研究的目的是评估用Mo Se ( f- cork) 的活性抗菌化合物生物功能化的软木对抑制大肠杆菌(Inh Ec ) 的影响。使用活力定量聚合酶链反应 (qPCR) 和通过选择性细胞活力 PMA 染色区分活细菌和死细菌,将来自大肠杆菌菌株的LacZ基因用作目标序列。为此,一个 2 7−4部分因子设计和生物过滤系统用于评估固定在颗粒状软木中的MoSe中的活性蛋白对InhEc 的影响。我们发现抗微生物活性的潜力随着f- cork 的增加而增加,从而有效地最大程度地减少细菌 (99.99%;p < 0.05)。对于暴露于 PMA 的细胞,用Mo Se功能化的f- cork对大肠杆菌生存力的影响分别为 0.024% 和 0.005%,这是处理 2 中的相关条件:(0 L/min)无通气,(5 %) Mo Se 和 (5 mm) 软木颗粒。总之,用Mo功能化的f- corkSe 具有生物吸附和抗菌特性,可有效减少大肠杆菌的生长。