scholarly journals Comparison of Total Culturable Virus Assay and Multiplex Integrated Cell Culture-PCR for Reliability of Waterborne Virus Detection

2004 ◽  
Vol 70 (6) ◽  
pp. 3632-3636 ◽  
Author(s):  
Hwa Kyung Lee ◽  
Yong Seok Jeong

ABSTRACT The total culturable virus assay (TCVA) and an integrated cell culture-PCR (ICC-PCR) were compared in parallel to evaluate their detection reliability. Source, finished, and tap water samples from three drinking water treatment plant systems were analyzed by TCVA, and every cell culture dish was subsequently examined by reverse transcription (RT) multiplex PCR using enterovirus- and adenovirus-specific primers. Twenty-seven of 180 (15%) inoculated dishes exhibited cytopathic effects (CPE). Virus concentrations for source water ranged from 3.3 to 21.0 most probable numbers of infectious units (MPN) per 100 liters. No finished or tap water samples were positive. On the other hand, 38 (21%) of the dishes were positive in multiplex ICC-PCR. Virus concentrations ranged from 4.5 to 10.2 MPN/100 liters for source water and 0 to 0.9 MPN/100 liters for finished and tap water. In spite of its superior sensitivity, the ICC-PCR assay resulted in lower virus concentration values than the TCVA for two of the source water sites. Retest of the CPE-positive dishes using reovirus-specific RT-PCR revealed that 24 of the 27 (89%) dishes were also positive for reoviruses. These observations suggested that the detection reliability of ICC-PCR is restricted by the primer sets that are integrated in the reaction mixture. The observation of an uneven distribution of PCR-positive culture dishes in a given sample raises an additional caution that simple extrapolation of the ICC-PCR result from the analysis of a limited fraction of collected samples should be avoided to minimize possible over- and underestimation of the amount of virus.

Author(s):  
Cristina Pignata ◽  
Silvia Bonetta ◽  
Sara Bonetta ◽  
Simone M. Cacciò ◽  
Anna R. Sannella ◽  
...  

The aim of this study was to evaluate the occurrence of Cryptosporidium oocysts in a drinking water treatment plant (DWTP) located in a rural area of northern Italy. Influent and effluent samples were collected at the DWTP over three years (2013–2016). In parallel, tap water samples from a public drinking fountain were collected as well. All samples were analyzed for the presence of Cryptosporidium spp. oocysts by a common method based on an immunomagnetic separation (IMS)/immunofluorescence assay (IFA), complemented by 4,6-diamidino-2-phenylindole (DAPI) staining. A reverse transcriptase-PCR (RT-PCR) protocol was added to evaluate oocyst viability. The results highlighted a high variability of oocyst concentrations across all samples (mean 4.3 ± 5.8/100 L) and a high variability in the percentage of DAPI-positive specimens (mean 48.2% ± 40.3%). Conversely, RT-PCR did not reveal the presence of viable C. parvum and C. hominis oocysts. A nested PCR targeting Cryptosporidium 18S ribosomal DNA, carried out in two water samples, confirmed the presence of a Cryptosporidium genotype associated with wild animals in the river and in tap water. The results obtained underline the vulnerability of the investigated surface water to Cryptosporidium spp. contamination. Although the recovered Cryptosporidium genotype is not a human pathogen, its presence demonstrates the existence of a potential pathogen Cryptosporidium spp. contamination risk. Moreover, these results underline the importance of also considering unconventional (not bacterial) biological contaminations (protozoa) in water resources in rural areas, including those of developed countries.


2020 ◽  
Vol 30 (Supplement_5) ◽  
Author(s):  
C Pignata ◽  
S i Bonetta ◽  
S a Bonetta ◽  
S M Cacciò ◽  
A R Sannella ◽  
...  

Abstract Over the past decade, several outbreaks of cryptosporidiosis have been reported in many EU countries in association with the consumption of contaminated drinking water, recreational waters, food consumption and contact with animals. The aim of this study was to evaluate the occurrence of Cryptosporidium oocysts in a drinking water treatment plant (DWTP) located in a rural area of northern Italy. Influent and effluent samples at the DWTP, together with tap water samples from a public fountain were collected for three years (2013-2016). All samples were analysed for the presence of Cryptosporidium spp. oocysts by the EPA Method 1623 based on immunomagnetic separation (IMS)/immunofluorescence assay (IFA), complemented by 4,6-diamidino-2-phenylindole (DAPI) staining. The detection has been implemented with a reverse transcriptase-PCR (RT-PCR) protocol to evaluate the oocyst viability. The results highlighted a high variability of oocyst concentrations in all samples (mean 4.3 - 5.8/100 L) and a high percentage variability of the DAPI-positive specimens (mean 48.2% - 40.3%). Conversely, RT-PCR did not reveal the presence of any viable C. parvum and C. hominis oocysts. A nested PCR targeting Cryptosporidium 18S ribosomal DNA was carried out in two water samples to deepen the understanding of the obtained results. This test revealed the presence of a particular Cryptosporidium genotype associated with wild animals in the river and in tap water. Although the recovered Cryptosporidium genotype is not a human pathogen, its presence demonstrates the existence of a potential pathogen Cryptosporidium spp. contamination risk. Moreover, these results underline the importance of considering unconventional (not bacterial) biological contaminations (protozoa) in water resources in rural areas, including those of developed countries. Key messages The supply source and the drinking water treatment plant examined represent a classic example of a system that requires continuous monitoring to guarantee population health. Cryptosporidium oocysts may still be a Public Health problem even in industrialized countries, especially in rural and mountain areas.


1989 ◽  
Vol 21 (3) ◽  
pp. 255-258 ◽  
Author(s):  
Evangélos Biziagos ◽  
Jacques Passagot ◽  
Jean-Marc Crance ◽  
Robert Deloince

The concentration of cell-culture-adapted hepatitis A virus (HAV) from experimentally contaminated distilled, drinking, waste and seawater was performed by using a filter adsorption-elu-tion method in the following conditions: HAV seeded in water was adsorbed at pH 4.0 to two nitrocellulose membranes (1.2 and 0.45 µm porosity for distilled and tap water or 8.0 and 3.0 µm porosity for waste and seawater), then eluted by 3% beef-extract at pH 8.5 and further concentrated by polyethylene glycol 6000 precipitation. Thus, HAV in 5 to 50 liters of seeded waters was concentrated approximately 1,700 to 17,000 fold with greater than 70% recovery of the initial virus added to the samples.


Author(s):  
Wenjin Xue ◽  
Christopher W. K. Chow ◽  
John van Leeuwen

Abstract The bacterial regrowth potential (BRP) method was utilised to indirectly measure the assimilable organic carbon (AOC) as an indicator for the assessment of the microbial regrowth potential in drinking water distribution systems. A model using various microbial growth parameters was developed in order to standardise the experimental interpretation for BRP measurement. This study used 82 experimental BRP data sets of water samples collected from the water treatment plant to locations (customer taps) in the distribution system. The data were used to model the BRP process (growth curve) by a data fitting procedure and to obtain a best-fitted equation. Statistical assessments and model validation for evaluating the equation obtained by fitting these 82 sets of data were conducted, and the results show average R2 values were 0.987 for treated water samples (collected at the plant prior to chlorination) and 0.983 for tap water (collected at the customer taps). The F values obtained from the F-test are all exceeded their corresponding F critical values, and the results from the t-test also showed a good outcome. These results indicate this model would be successfully applied in modelling BRP in drinking water supply systems.


2019 ◽  
Vol 19 (7) ◽  
pp. 2098-2106
Author(s):  
Chelsea W. Neil ◽  
Yingying Zhao ◽  
Amy Zhao ◽  
Jill Neal ◽  
Maria Meyer ◽  
...  

Abstract Source water quality can significantly impact the efficacy of water treatment unit processes and the formation of chlorinated and brominated trihalomethanes (THMs). Current water treatment plant performance models may not accurately capture how source water quality variations, such as organic matter variability, can impact treatment unit processes. To investigate these impacts, a field study was conducted wherein water samples were collected along the treatment train for 72 hours during a storm event. Systematic sampling and detailed analyses of water quality parameters, including non-purgeable organic carbon (NPOC), UV absorbance, and THM concentrations, as well as chlorine spiking experiments, reveal how the THM formation potential changes in response to treatment unit processes. Results show that the NPOC remaining after treatment has an increased reactivity towards forming THMs, and that brominated THMs form more readily than chlorinated counterparts in a competitive reaction. Thus both the reactivity and quantity of THM precursors must be considered to maintain compliance with drinking water standards, a finding that should be incorporated into the development of model-assisted treatment operation and optimization. Advanced granular activated carbon (GAC) treatment beyond conventional coagulation–flocculation–sedimentation processes may also be necessary to remove the surge loading of THM-formation precursors during a storm event.


2009 ◽  
Vol 36 (6) ◽  
pp. 1095-1106 ◽  
Author(s):  
Heather P. Sim ◽  
Donald H. Burn ◽  
Bryan A. Tolson

Source water protection involves safeguarding water supplies from contamination and depletion. Despite best efforts, spills cannot always be prevented from entering a source water body. However, many spills can be prevented from entering a drinking water treatment plant if an early warning source water monitoring station is used. These stations provide downstream water utilities with advanced notification of spills so the utilities have time to implement their responses. This paper addresses the design of an early warning monitoring station for a riverine source of drinking water. Riverine water supplies face many threats related to accidental spills, which are inherently uncertain in nature. Therefore, designing a monitoring station for the detection of these events requires a probabilistic modelling approach. The design objectives include maximizing the probabilities of detection and of having a threshold amount of warning time. The methodology is applied to a water supply intake on the Grand River in southern Ontario.


2005 ◽  
Vol 71 (2) ◽  
pp. 1042-1050 ◽  
Author(s):  
Gerald Sedmak ◽  
David Bina ◽  
Jeffrey MacDonald ◽  
Lon Couillard

ABSTRACT Reoviruses, enteroviruses, and adenoviruses were quantified by culture for various ambient waters in the Milwaukee area. From August 1994 through July 2003, the influent and effluent of a local wastewater treatment plant (WWTP) were tested monthly by a modified U.S. Environmental Protection Agency Information Collection Rule (ICR) organic flocculation cell culture procedure for the detection of culturable viruses. Modification of the ICR procedure included using Caco-2, RD, and HEp-2 cells in addition to BGM cells. Lake Michigan source water for two local drinking water treatment plants (DWTPs) was also tested monthly for culturable viruses by passing 200 liters of source water through a filter and culturing a concentrate representing 100 liters of source water. Reoviruses, enteroviruses, and adenoviruses were detected frequently (105 of 107 samples) and, at times, in high concentration in WWTP influent but were detected less frequently (32 of 107 samples) in plant effluent and at much lower concentrations. Eighteen of 204 samples (8.8%) of source waters for the two DWTPs were positive for virus and exclusively positive for reoviruses at relatively low titers. Both enteroviruses and reoviruses were detected in WWTP influent, most frequently during the second half of the year.


Author(s):  
Zhiquan Liu ◽  
Yongpeng Xu ◽  
Yuan Wang ◽  
Fuyi Cui

Abstract Quinolone (QN) antibiotics are widely used all over the world and have been frequently detected in source water, but the occurrence in tap water and the treatment efficiencies of QNs by drinking-water treatment plants (DWTPs) were rarely reported. In the present study, the occurrence and distribution of six representative QNs in three urban DWTPs of China were investigated. The results showed that the concentrations of total QNs in the three source waters ranged from 26.4 ng/L to 313.8 ng/L and all of the six QNs were detectable with a detection frequency of 100% (4.6 to 121.7 ng/L). Enrofloxacin (ENR) and ofloxacin (OFL) were the dominant species of QNs and accounted for 40.1% to 79.5% of the total QNs. After the treatments, there were still considerable QNs in the finished water (total amounts of 74.9 ng/L to 148.4 ng/L). The adsorbed QNs could be readily treated with the removal of turbidity by DWTPs, but only a part of the dissolved QNs (13.6% to 68.5%) can be removed. This implies that the dissolved QNs were more hazardous in the source water. Pre-oxidation and disinfection could remove 15.8 ± 8.3% and 16.9 ± 10.8% of dissolved QNs, respectively, depending on the chemical structure of QNs and the types of oxidant. Chemical oxidation was more efficient than coagulation-sedimentation and filtration for the treatment of dissolved QNs. Ozone-granular activated carbon filtration may fail to remove dissolved QNs in the actual DWTPs, because of the insufficient dosage of oxidant and the competition effect of natural organic matter.


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