Use of enzymatic methods for rapid enumeration of coliforms in freshwaters

George, Isabelle; Petit, Muriel; Servais, Pierre

doi:10.1046/j.1365-2672.2000.00977.x

Cited by 90 publications

(99 citation statements)

References 31 publications

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“…GAL and GUD properties of TC and E. coli have been exploited on freshwater (Berg and Fiksdal 1988;Tryland and Fiksdal 1998;George et al 2000;Fiksdal and Tryland 2008) and seawater (Davies et al 1995) …”

Section: Application Of Rapid Enzymatic Assays For Bacterial Detectionmentioning

confidence: 99%

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A biosensor platform for rapid detection of E. coli in drinking water

Hesari

Alum

El-Zein

et al. 2016

Enzyme and Microbial Technology

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The need for rapid, specific and sensitive assays that provide a detection of bacterial indicators are important for monitoring water quality. Rapid detection using biosensor is a novel approach for microbiological testing applications. Besides, validation of rapid methods is an obstacle in adoption of such new bio-sensing technologies. In this study, the strategy developed is based on using the compound 4-methylumbelliferyl glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli β-D-glucuronidase (GUD) enzyme to yield a fluorogenic product that can be quantified and directly related to the number of E. coli cells present in water samples.The detection time required for the biosensor response ranged from 30 to 120 minutes, depending on the number of bacteria. The specificity of the MUG based biosensor platform assay for the detection of E. coli was examined by pure cultures of non-target bacterial genera and also non-target substrates. GUD activity was found to be specific for E. coli and no such enzymatic activity was detected in other species. Moreover, the sensitivity of rapid enzymatic assays was investigated and repeatedly determined to be less than 10 E. coli cells per reaction vial concentrated from 100 mL of water samples.The applicability of the method was tested by performing fluorescence assays under pure and mixed bacterial flora in environmental samples. In addition, the procedural QA/QC for routine monitoring of drinking water samples have been validated by comparing the performance of the biosensor platform for the detection of E. coli and culture-based standard techniques such as Membrane Filtration (MF). The results of this study ii indicated that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. The procedural QA/QC of the biosensor will provide both industry and regulatory authorities a useful tool for near real-time monitoring of E. coli in drinking water samples. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water.

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Section: Application Of Rapid Enzymatic Assays For Bacterial Detectionmentioning

confidence: 99%

“…Another limitation of these techniques is the inability to detect viable but non-culturable bacteria (George et al 2000). Hence, rapid, sensitive and specific assays that provide a near real-time detection of bacterial indicators are of primary importance for monitoring microbiological quality of water.…”

mentioning

confidence: 99%

A biosensor platform for rapid detection of E. coli in drinking water

Hesari

Alum

El-Zein

et al. 2016

Enzyme and Microbial Technology

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“…If clusters of aggregated or particle bound E. coli are present in a sample, culture based methods count clusters as single units thus underestimating the number of cells present whereas enzyme assays account for the activity of each cell thus giving a better representation of the total number of cells present. For these reasons it is difficult to correlate the two approaches although this is commonly done due to a lack of a practical alternative standard reference method [9,12,16]. There is a growing body of evidence pointing to the virulence of VBNC bacteria and suggesting the importance of measuring their numbers in environmental waters [17][18][19].…”

Section: Introductionmentioning

confidence: 99%

“…Garcia-Armisen [12] using a method from George [16] based on 4-MUG, reports GUS activities of approximately 100 fmol (4-MUG) min 1 per culturable E. coli for lightly contaminated freshwater samples (i.e. 100-1000 E. coli.…”

Section: Introductionmentioning

confidence: 99%

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ColiSense, today's sample today: A rapid on-site detection of β-d-Glucuronidase activity in surface water as a surrogate for E. coli

Heery

Briciu-Burghina

Zhang

et al. 2016

Talanta

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ab s t r a c t A sensitive field-portable fluorimeter with incubating capability and triplicate sample chambers was designed and built. The system was optimised for the on-site analysis of E. coli in recreational waters using fluorescent based enzyme assays. The target analyte was β-D-Glucuronidase (GUS) which hydrolyses a synthetic substrate 6-Chloro-4-Methyl-Umbelliferyl-β-D-Glucuronide (6-CMUG) to release the fluorescent molecule 6-Chloro-4-Methyl-Umbelliferyl (6-CMU). The system was calibrated with 6-CMU standards. A LOD of 5 nM and a resolution of less than 1 nM was determined while enzyme kinetic tests showed detection of activities below 1 pmol min 1 mL 1 of sample. A field portable sample preparation, enzyme extraction protocol and continuous assay were applied with the system to analyse freshwater and marine samples. Results from a one day field trial are shown which demonstrated the ability of the system to deliver results on-site within a 75 min period.

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Assessment of Presence of Heavy Metals and Other Pollution Burden Parameters and Their Effect on Water Quality in Benin City, Edo State

Adams

Ogedegbe²,

Tawari–Fufeyin

2016

Environmental Quality Management

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An assessment of the heavy metal content and biological pollution burden of an abattoir dumpsite, an automobile carwash, the Ikpoba River, and a private water borehole located near the abattoir dumpsite was conducted in Benin City in Southern Nigeria, during the rainy and dry seasons. The water samples’ pH was lower than the permissible values, with the river water displaying color and possessing a slight odor. Iron concentrations exceeded the permissible limits for drinking water at all four of the sampling stations in the study area, although not in samples collected from the control station, station 5, a public water borehole that is located some distance away. Lead values were higher than drinking water standards in the samples collected from stations 1 through 4, with some exceptions during the rainy season. Copper, nickel, arsenic, chromium, and aluminum were detected in three out of the four sampling stations located within the study area. However, these metals were detected at levels that were within regulatory limits. The detection of three metals (iron, lead, and zinc) within the study area at levels exceeding drinking water limits suggested that a complete cycle of movement of pollutants into the sampling stations had occurred. Significant biological presences were also detected, as the Ikpoba River water had a standard plate count (SPC) of 130 most probable number (MPN) and a 22 MPN presumptive coliform count (PCC) or 22 MPN. However, Escherichia coli (E. coli) confirmatory tests recorded less than 2 MPN in both seasons. The study has shown that the quality of water in the river may be compromised by effluent discharges from the dumpsite and the carwash channel. Therefore, this situation requires more stringent enforcement of local environmental laws and maintenance of safe distances between domestic and industrial waste sources and domestic or private water wells.

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Use of enzymatic methods for rapid enumeration of coliforms in freshwaters

Cited by 90 publications

References 31 publications

A biosensor platform for rapid detection of E. coli in drinking water

A biosensor platform for rapid detection of E. coli in drinking water

ColiSense, today's sample today: A rapid on-site detection of β-d-Glucuronidase activity in surface water as a surrogate for E. coli

Assessment of Presence of Heavy Metals and Other Pollution Burden Parameters and Their Effect on Water Quality in Benin City, Edo State

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