d8073a448ed5f06763d6614f097b5923 2021_Infante_et_al_Water.pdf 1e278a524c3ad2ccaa838d64a4effac3ca35f785 2021_Infante_et_al_Water.pdf a55208bc9c89515318fe6da0795ac9a62c13d5dc6faab1c3dfa84224218ee1e6 2021_Infante_et_al_Water.pdf Title: Biofunctionalization of Cork with Moringa oleifera Seeds and Use of PMA Staining and qPCR to Detect Viability of Escherichia coli Subject: 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. Keywords: cork-based biofilter; Escherichia coli; viability qPCR; Lac Z gene; propidium monoazide; disinfection; antimicrobial activity Author: Nury Infante, Refugio Rodríguez, Yaneth Bartolo, Olga Sánchez, Isabel Sanz, Lizeth Bermeo and Jordi Morató Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sat Oct 2 10:06:44 2021 CEST ModDate: Mon Oct 11 10:21:55 2021 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1130533 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- MWHIBA+VnURWPalladioL Type 1 Custom yes yes no 10 0 ORLLYG+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 KPNRTR+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 GOQDSB+URWPalladioL-BoldItal Type 1 Custom yes yes no 24 0 NVFSXO+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 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FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 170 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 176 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 182 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 188 0 FNTSBS+TimesNewRomanPS-ItalicMT CID TrueType Identity-H yes yes yes 194 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 205 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 211 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 217 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-01-18 02:30:58 CET RepresentationInformation: 2021_Infante_et_al_Water.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-01-14 01:34:21 CET Size: 1130533 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 436 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Bacterial Growth Kinetic and Selection of the Reporter Gene Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Genomic DNA Extraction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Real-Time PCR for Detection of E. coli Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Children: Item: Title: LacZ Gene Amplification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: LacZ Gene Standard Curves Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Quantitative Real-Time PCR Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Sample Concentration, PMA Treatment and Relevant Test Controls Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Live/Dead Cell Viability Assays Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Functionalization of Cork with MoSe Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Biofiltration System with Filter Cartridges Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Fractional 27-4 Experimental Design Using Cork as Filter Material Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Antimicrobial Activity of Cork Functionalized with MoSe Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Children: Item: Title: Growth Kinetic of E. coli and Amplification of LacZ Gen Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Functionalization of Cork with MoSe and Effect on InhEc Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: LacZ vqPCR Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Live/Dead Bacterial Cell Viability Test Using Cork as Filter Material Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Children: Item: Title: Growth Kinetic of E. coli and Amplification of LacZ Gene Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Functionalization of Cork with MoSe and InhEc Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: LacZ vqPCR Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Live/Dead Bacterial Cell Viability Test Using Cork as Filter Material Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Info: Title: Biofunctionalization of Cork with Moringa oleifera Seeds and Use of PMA Staining and qPCR to Detect Viability of Escherichia coli Author: Nury Infante, Refugio Rodríguez, Yaneth Bartolo, Olga Sánchez, Isabel Sanz, Lizeth Bermeo and Jordi Morató Subject: 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. 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