scholarly journals Photoelectrocatalytic disinfection of water and wastewater: performance evaluation by qPCR and culture techniques

2012 ◽  
Vol 11 (1) ◽  
pp. 21-29 ◽  
Author(s):  
Danae Venieri ◽  
Efthalia Chatzisymeon ◽  
Eleonora Politi ◽  
Spiridon S. Sofianos ◽  
Alexandros Katsaounis ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Charge Carriers ◽  
Treated Wastewater ◽  
Bacterial Inactivation ◽  
Culture Techniques ◽  
E Coli ◽  
Log Reduction ◽  
Water Matrix ◽  
Water And Wastewater ◽  
Polymerase Chain

Photoelectrocatalytic oxidation (PEC) was evaluated as a disinfection technique using water and secondary treated wastewater spiked with Escherichia coli and Enterococcus faecalis. PEC experiments were carried out using a TiO2/Ti-film anode and a zirconium cathode under simulated solar radiation. Bacterial inactivation was monitored by culture and quantitative polymerase chain reaction (qPCR). Inactivation rates were enhanced when the duration of the treatment was prolonged and when the bacterial density and the complexity of the water matrix were decreased. E. coli cells were reduced by approximately 6 orders of magnitude after 15 min of PEC treatment in water at 2V of applied potential and an initial concentration of 107 CFU/mL; pure photocatalysis (PC) led to about 5 log reduction, while electrochemical oxidation alone resulted in negligible inactivation. The superiority of PEC relative to PC can be attributed to a more efficient separation of the photogenerated charge carriers. Regarding disinfection in mixed bacterial suspensions, E. coli was more susceptible than E. faecalis at a potential of 2V. The complex composition of wastewater affected disinfection efficiency, yielding lower inactivation rates compared to water treatment. qPCR yielded lower inactivation rates at longer treatment times than culture techniques, presumably due to the fact that the latter do not take into account the viable but not culturable state of microorganisms.

scholarly journals Simultaneous removal of estrogens and pathogens from secondary treated wastewater by solar photocatalytic treatment

2014 ◽  
Vol 16 (3) ◽  
pp. 543-552 ◽  
Keyword(s):  
Solar Radiation ◽  
Estrogenic Activity ◽  
Treated Wastewater ◽  
Simultaneous Removal ◽  
E Coli ◽  
Water Matrix ◽  
Treated Effluent ◽  
Emerging Compounds ◽  
Wastewater Disinfection ◽  
Water And Wastewater

<p>Recently, the fate of emerging compounds in environmentally relevant samples has attracted considerable attention. Solar semiconductor photocatalysis may offer an appealing methodology to treat such contaminants. At the same time the use of solar photocatalysis for water and wastewater disinfection is a topic well-documented in the literature. In this respect, the simultaneous degradation of synthetic estrogen 17α-ethynylestradiol (EE2) and <em>Escherichia coli </em>removal employing simulated solar radiation and TiO<sub>2</sub> as the photocatalyst was investigated. In general, the more complex the water matrix is the slower <em>E. coli</em> removal becomes, while the presence of <em>E. coli</em> in the reaction mixture did not obstruct EE2 removal.</p> <div> <p>Although EE2 removal occurred relatively fast, overall estrogenic activity was only partially removed. This implies that other species inherently present in the effluent and/or some photocatalytic transformation by-products may be proportionately more estrogenic than EE2. Overall, the use of solar radiation can constitute an advantageous treatment strategy for the simultaneous removal of micro-pollutants and pathogens from secondary treated effluent.</p> </div> <p>&nbsp;</p>

scholarly journals Eliminating false positives in a qPCR assay for the detection of the uidA gene in Escherichia coli

2013 ◽  
Vol 11 (3) ◽  
pp. 382-386 ◽  
Author(s):  
Richard Kibbee ◽  
Natalie Linklater ◽  
Banu Örmeci
Keyword(s):  
Escherichia Coli ◽  
Quantitative Polymerase Chain Reaction ◽  
Qpcr Assay ◽  
Uida Gene ◽  
Chain Reaction ◽  
Taq Polymerase ◽  
E Coli ◽  
Gene Copies ◽  
Polymerase Chain ◽  
Low Levels

Due to contaminant Escherichia coli DNA present in recombinant Taq polymerase reagents, it is not possible to reliably detect low levels of E. coli in samples using the quantitative polymerase chain reaction (qPCR) assay. Native Taq polymerase was successfully used in this study to detect five uidA gene copies (5 fg of genomic DNA) of the uidA gene.

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in Cranberry, Lemon, and Lime Juice Concentrates

2003 ◽  
Vol 66 (9) ◽  
pp. 1637-1641 ◽  
Author(s):  
MARA C. L. NOGUEIRA ◽  
OMAR A. OYARZÁBAL ◽  
DAVID E. GOMBAS
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Escherichia Coli O157 ◽  
Bacterial Inactivation ◽  
Lime Juice ◽  
E Coli ◽  
Log Reduction ◽  
C 32

The production of thermally concentrated fruit juices uses temperatures high enough to achieve at least a 5-log reduction of pathogenic bacteria that can occur in raw juice. However, the transportation and storage of concentrates at low temperatures prior to final packaging is a common practice in the juice industry and introduces a potential risk for postconcentration contamination with pathogenic bacteria. The present study was undertaken to evaluate the likelihood of Escherichia coli O157: H7, Listeria monocytogenes and Salmonella surviving in cranberry, lemon, and lime juice concentrates at or above temperatures commonly used for transportation or storage of these concentrates. This study demonstrates that cranberry, lemon, and lime juice concentrates possess intrinsic antimicrobial properties that will eliminate these bacterial pathogens in the event of postconcentration recontamination. Bacterial inactivation was demonstrated under all conditions; at least 5-log Salmonella inactivation was consistently demonstrated at −23°C (−10°F), at least 5-log E. coli O157:H7 inactivation was consistently demonstrated at −11°C (12°F), and at least 5-log L. monocytogenes inactivation was consistently demonstrated at 0°C (32°F).

scholarly journals Use of Fluorescence Quantitative Polymerase Chain Reaction (PCR) for the Detection of Escherichia coli Adhesion to Pig Intestinal Epithelial Cells

2016 ◽  
Vol 19 (3) ◽  
pp. 619-625 ◽  
Author(s):  
C.H. Dai ◽  
L.N. Gan ◽  
W.U. Qin ◽  
C. Zi ◽  
G.Q. Zhu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Relative Number ◽  
Intestinal Epithelial ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

AbstractAn efficient and accurate method to testEscherichia coli(E. coli) adhesion to intestinal epithelial cells will contribute to the study of bacterial pathogenesis and the function of genes that encode receptors related to adhesion. This study used the quantitative real-time polymerase chain reaction (qPCR) method. qPCR primers were designed from thePILINgene ofE. coliF18ab, F18ac, and K88ac, and the pig β-ACTINgene. Total deoxyribonucleic acid (DNA) fromE. coliand intestinal epithelial cells (IPEC-J2 cells) were used as templates for qPCR. The 2−ΔΔCtformula was used to calculate the relative number of bacteria in cultures of different areas. We found that the relative numbers of F18ab, F18ac, and K88ac that adhered to IPEC-J2 cells did not differ significantly in 6-, 12-, and 24-well culture plates. This finding indicated that there was no relationship between the relative adhesion number ofE. coliand the area of cells, so the method of qPCR could accurately test the relative number ofE. coli. This study provided a convenient and reliable testing method for experiments involvingE. coliadhesion, and also provided innovative ideas for similar detection methods.

scholarly journals Environmental inactivation and irrigation-mediated regrowth of Escherichia coli O157:H7 on romaine lettuce when inoculated in a fecal slurry matrix

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6591 ◽  
Author(s):  
Jennifer A. Chase ◽  
Melissa L. Partyka ◽  
Ronald F. Bond ◽  
Edward R. Atwill
Keyword(s):  
Escherichia Coli ◽  
Field Trials ◽  
Inactivation Rate ◽  
Bacterial Inactivation ◽  
Romaine Lettuce ◽  
E Coli ◽  
Log Reduction ◽  
Salinas Valley ◽  
Kinetics Of ◽  
Insight Into

Field trials were conducted in July–August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.

Disinfection of tertiary wastewater effluent prior to river discharge using peracetic acid; treatment efficiency and results on by-products formed in full scale tests

2013 ◽  
Vol 68 (8) ◽  
pp. 1852-1856 ◽  
Author(s):  
Per Overgaard Pedersen ◽  
Erling Brodersen ◽  
David Cecil
Keyword(s):  
Peracetic Acid ◽  
Full Scale ◽  
Treated Wastewater ◽  
Decomposition Products ◽  
Sand Filter ◽  
E Coli ◽  
Log Reduction ◽  
Before And After ◽  
Full Scale Tests ◽  
P Removal

This is an investigation of chemical disinfection, with peracetic acid (PAA), in a tertiary sand filter at a full scale activated sludge plant with nitrification/denitrification and P-removal. The reduction efficiency of Escherichia coli and intestinal enterococci in the sand filter is reported. E. coli log reductions of between 0.4 and 2.2 were found with contact times from 6 to 37 min and with dosing from 0 to 4.8 mg L−1. The average log reduction was 1.3. The decomposition products, bromophenols, chlorophenols and formaldehyde and residual H2O2 were measured before and after the sand filter. The residual H2O2 concentration in the effluent was critical at short contact times and high dosages of PAA due to the discharge limit of 25 μg L−1. The other three products could not be detected at 0.1 μg L−1 levels. The chemical cost of PAA dosing is estimated to be 0.039 US$ m−3 treated wastewater.

scholarly journals Antibiotic Resistance and Sewage-Associated Marker Genes in Untreated Sewage and a River Characterized During Baseflow and Stormflow

2021 ◽  
Vol 12 ◽  
Author(s):  
Warish Ahmed ◽  
Pradip Gyawali ◽  
Kerry A. Hamilton ◽  
Sayalee Joshi ◽  
David Aster ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Water Samples ◽  
Quantitative Polymerase Chain Reaction ◽  
River System ◽  
Marker Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Untreated Sewage ◽  
Polymerase Chain

Since sewage is a hotspot for antibiotic resistance genes (ARGs), the identification of ARGs in environmental waters impacted by sewage, and their correlation to fecal indicators, is necessary to implement management strategies. In this study, sewage treatment plant (STP) influent samples were collected and analyzed using quantitative polymerase chain reaction (qPCR) to investigate the abundance and correlations between sewage-associated markers (i.e., Bacteroides HF183, Lachnospiraceae Lachno3, crAssphage) and ARGs indicating resistance to nine antibiotics (belonging to aminoglycosides, beta-lactams, sulfonamides, macrolides, and tetracyclines). All ARGs, except blaVIM, and sewage-associated marker genes were always detected in untreated sewage, and ermF and sul1 were detected in the greatest abundances. intl1 was also highly abundant in untreated sewage samples. Significant correlations were identified between sewage-associated marker genes, ARGs and the intl1 in untreated sewage (τ = 0.488, p = 0.0125). Of the three sewage-associated marker genes, the BIO-ENV procedure identified that HF183 alone best maximized correlations to ARGs and intl1 (τ = 0.590). Additionally, grab samples were collected from peri-urban and urban sites along the Brisbane River system during base and stormflow conditions, and analyzed for Escherichia coli, ARGs, the intl1, and sewage-associated marker genes using quantitative polymerase chain reaction (qPCR). Significant correlations were identified between E. coli, ARGs, and intl1 (τ = 0.0893, p = 0.0032), as well as with sewage-associated marker genes in water samples from the Brisbane River system (τ = 0.3229, p = 0.0001). Of the sewage-associated marker genes and E. coli, the BIO-ENV procedure identified that crAssphage alone maximized correlations with ARGs and intl1 in river samples (τ = 0.4148). Significant differences in E. coli, ARGs, intl1, and sewage-associated marker genes, and by flow condition (i.e., base vs. storm), and site types (peri-urban vs. urban) combined were identified (R = 0.3668, p = 0.0001), where percent dissimilarities between the multi-factorial groups ranged between 20.8 and 11.2%. Results from this study suggest increased levels of certain ARGs and sewage-associated marker genes in stormflow river water samples compared to base flow conditions. E. coli, HF183 and crAssphage may serve as potential indicators of sewage-derived ARGs under stormflow conditions, and this merits further investigation. Data presented in this study will be valuable to water quality managers to understand the links between sewage pollution and ARGs in urban environments.

scholarly journals Rapid and Accurate Detection of Bacteriophage Activity againstEscherichia coliO157:H7 by Propidium Monoazide Real-Time PCR

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Hui Liu ◽  
Yan D. Niu ◽  
Jinquan Li ◽  
Kim Stanford ◽  
Tim A. McAllister
Keyword(s):  
Cell Surface ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Extracellular Dna ◽  
Bacterial Cells ◽  
Propidium Monoazide ◽  
Direct Plating ◽  
E Coli ◽  
Polymerase Chain ◽  
Phage Activity

Conventional methods to determine the efficacy of bacteriophage (phage) for biocontrol ofE. colirequire several days, due to the need to culture bacteria. Furthermore, cell surface-attached phage particles may lyse bacterial cells during experiments, leading to an overestimation of phage activity. DNA-based real-time quantitative polymerase chain reaction (qPCR) is a fast, sensitive, and highly specific means of enumerating pathogens. However, qPCR may underestimate phage activity due to its inability to distinguish viable from nonviable cells. In this study, we evaluated the suitability of propidium monoazide (PMA), a microbial membrane-impermeable dye that inhibits amplification of extracellular DNA and DNA within dead or membrane-compromised cells as a means of using qPCR to identify only intactE. colicells that survive phage exposure.Escherichia coliO157:H7 strain R508N and 4 phages (T5-like, T1-like, T4-like, and O1-like) were studied. Results compared PMA-qPCR and direct plating and confirmed that PMA could successfully inhibit amplification of DNA from compromised/damaged cellsE. coliO157:H7. Compared to PMA-qPCR, direct plating overestimated (P< 0.01) phage efficacy as cell surface-attached phage particles lysedE. coliO157:H7 during the plating process. Treatment of samples with PMA in combination with qPCR can therefore be considered beneficial when assessing the efficacy of bacteriophage for biocontrol ofE. coliO157:H7.

scholarly journals Plant-Derived Antimicrobials ReduceE. coliO157:H7 Virulence Factors Critical for Colonization in Cattle Gastrointestinal TractIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Sangeetha Ananda Baskaran ◽  
Kumar Venkitanarayanan
Keyword(s):  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Rumen Fluid ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
E Coli ◽  
Intestinal Contents ◽  
Polymerase Chain

This study investigated the effect of subinhibitory concentrations (SIC) of five plant-derived antimicrobials (PDAs), namely, trans cinnamaldehyde, eugenol, carvacrol, thymol, andβ-resorcylic acid, onE. coliO157:H7 (EHEC) attachment and invasion of cultured bovine colonic (CO) and rectoanal junction (RAJ) epithelial cells. In addition, PDAs’ effect on EHEC genes critical for colonization of cattle gastrointestinal tract (CGIT) was determined in bovine rumen fluid (RF) and intestinal contents (BICs). Primary bovine CO and RAJ epithelial cells were established and were separately inoculated with three EHEC strains with or without (control) SIC of each PDA. Following incubation, EHEC that attached and invaded the cells were determined. Furthermore, the expression of EHEC genes critical for colonization in cattle was investigated using real-time, quantitative polymerase chain reaction in RF and BICs. All the PDAs decreased EHEC invasion of CO and RAJ epithelial cells (P<0.05). The PDAs also downregulated (P<0.05) the expression of EHEC genes critical for colonization in CGIT. Results suggest that the PDAs could potentially be used to control EHEC colonization in cattle; however follow-upin vivostudies in cattle are warranted.

Inactivation of Cryptosporidium parvum oocysts and other microbes in water and wastewater by electrochemically generated mixed oxidants

2000 ◽  
Vol 41 (7) ◽  
pp. 127-134 ◽  
Author(s):  
M. J. Casteel ◽  
M. D. Sobsey ◽  
M. J. Arrowood
Keyword(s):  
Escherichia Coli ◽  
Cell Culture ◽  
Cryptosporidium Parvum ◽  
Propidium Iodide ◽  
Treated Wastewater ◽  
E Coli ◽  
Water And Wastewater ◽  
Cryptosporidium Parvum Oocysts ◽  
Cell Culture Assay ◽  
Escherichia Coli B

Alternative disinfectants of water and wastewater are needed because conventional chlorination is ineffective against C. parvum oocysts. Reliable indicators of disinfection efficacy against C. parvum also are needed. Mixedoxidants (MO) electrochemically generated from brine were evaluated in batch disinfection experiments for inactivation of C. parvum oocysts and Cl. perfringensspores in both oxidant demand-free (ODF) water and treated wastewater. Coliphage MS2 and Escherichia coli B were also tested under some conditions. C. parvum oocyst infectivity was quantified by cell culture assay, and the dyes DAPI (4′,6-diamidino-2-phenylindole) and propidium iodide (PI) were used to assess oocyst viability in wastewater experiments. In treated wastewater dosed with 10–13 mg/L MO, inactivation after 90 minutes was about 3 log10 for C. parvum and about 2.5 log10 for Cl. perfringens spores; MS2 and E. coli were rapidly inactivated by &gt; 5 log10. In ODF water, a 4 mg/L dose of MO inactivated ∼3 log10 of C. parvum oocysts and ∼1.5 log10 of Cl. perfringens spores. Inactivation of C. parvum oocysts and Cl. perfringensspores was less extensive at a lower MO dose of 2 mg/L. The use of DAPI and PI to determine viability of oocysts treated with MO did not correlate with, and greatly overestimated, cell culture infectivity. At practical doses and contact times, MO disinfection of water and wastewater achieves appreciable inactivation of both C. parvum oocysts and Cl. perfringens spores. Cl. perfringens spores reliably indicated oocyst inactivation by MO, but E. coli and coliphage MS2 were inactivated much too rapidly to indicate C. parvum inactivation.

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