Protocol for the recovery and detection of Escherichia coli in environmental water samples

Briciu-Burghina, Ciprian; Heery, Brendan; Regan, Fiona

doi:10.1016/j.aca.2017.02.035

Cited by 21 publications

(18 citation statements)

References 34 publications

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“…Using the proposed procedure, it is possible to monitor indirectly the presence and distribution of E. coli in seawater. Compared to other technologies developed for the rapid detection of bacteria in recreational waters, the developed protocol offers also the advantages of working with low volumes of sample, disposable microplates, and simple instrumentation, therefore it is cheaper than other commercial diagnostic kits for E. coli detection based on molecular approaches [22][23].…”

Section: Discussionmentioning

confidence: 99%

β-glucuronidase activity determination as an indirect estimate of Escherichia coli: development of a miniaturized assay and its application to seawater samples

Caruso¹

2017

J Clin Microbiol Biochem Technol

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DOICC By 046Citation: Caruso G, Caruso R, Monticelli LS, Maimone G, Crisafi E (2017) -glucuronidase activity determination as an indirect estimate of Escherichia coli:development of a miniaturized assay and its application to seawater samples. J Clin Microbiol Biochem Technol 3(3): 046-050. Clinical Group AbstractBackground: The search of rapid methods for the detection of Escherichia coli in coastal marine waters is a topic of scientifi c interest to evaluate potential risks to human health related to their low bacteriological quality.The context and purpose of the study: A miniaturized assay for the analytical determination of β− glucuronidase activity in seawater as a selective marker of Escherichia coli was developed by using the chromogenic Indoxyl-β-D-glucuronide (IBDG) substrate. This compound is specifi cally cleaved by E. coli, releasing an insoluble chromophoric blue-indigo product that precipitates and is measured at 450 nm wavelength by a microplate reader. Results:After its preliminary optimization, the enzymatic assay was applied to the analysis of seawater samples and enabled to discriminate them according to their pollution level. Main fi ndings:The fi rst obtained data proved the suitability of the developed miniaturized enzymatic assay for fecal contamination monitoring. It can be used for the detection and indirect quantifi cation of E. coli, without the need for confi rmatory steps. Conclusions:This study suggests that the proposed analytical protocol is suitable for E. coli monitoring in seawater, and provides in a short time (i.e. 2 hours from sampling) results which are in agreement with the standard culture counts. Brief summary:The results obtained with the developed IBDG protocol encourage its use for environmental quality assessment. Any potential implications:The possibility to obtain near "real-time" data on the occurrence and distribution of anthropogenic inputs makes this method a simple and quick tool for early warning detection of marine fecal pollution.

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Section: Discussionmentioning

confidence: 99%

β-glucuronidase activity determination as an indirect estimate of Escherichia coli: development of a miniaturized assay and its application to seawater samples

Caruso¹

2017

J Clin Microbiol Biochem Technol

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“…While high-throughput developments enhance their processing, the processes remain lengthy because samples are collected and sent to laboratories for results to be provided within 18-24 h. This reduces effectiveness and contributes to reactive management when high-concentration events occur [22,39,41,42]. Unreliability is another issue, as CBM are often unable to provide reliable analyses of samples when the samples contain several different bacterial organisms or sediment levels such as in marine waters with high sediment levels, often returning false negatives and underestimations when the concentration of E. coli is low [22,43,44]. As a result, other detection techniques must be deployed to analyse complex samples [39,45,46].…”

Section: Culture-based Assaysmentioning

confidence: 99%

“…This was confirmed by Guo et al [32] when their CBM tests found that only <3% of the total bacterial population in their water sample was culturable. Additionally, bacterial load is often underestimated because only viable organisms are detected, excluding VBNC [22,44].…”

Section: Culture-based Assaysmentioning

confidence: 99%

“…A preparation method assay protocol developed by Briciu-Burghina et al [44] has a result time of 75 min for the detection of E. coli using GUS activity. The two main steps are (i) sample preparation using syringe filters with a dual purpose (the recovery and preconcentration of E. coli) and (ii) measuring of GUS activity.…”

Section: IIImentioning

confidence: 99%

“…Syringe filtration and recovery are conducted onsite. The incubation of the lysing syringe filter requires a portable incubator, and detection requires a handheld portable fluorimeter [44]. (1) This method is not automated, but it is also not complex (filtering, incubation, and sample placement).…”

Section: IIImentioning

confidence: 99%

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Monitoring Approaches for Faecal Indicator Bacteria in Water: Visioning a Remote Real-Time Sensor for E. coli and Enterococci

Offenbaume

Bertone

Stewart

2020

Water

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A comprehensive review was conducted to assess the current state of monitoring approaches for primary faecal indicator bacteria (FIB) E. coli and enterococci. Approaches were identified and examined in relation to their accuracy, ability to provide continuous data and instantaneous detection results, cost, environmental awareness regarding necessary reagent release or other pollution sources, in situ monitoring capability, and portability. Findings showed that several methods are precise and sophisticated but cannot be performed in real-time or remotely. This is mainly due to their laboratory testing requirements, such as lengthy sample preparations, the requirement for expensive reagents, and fluorescent tags. This study determined that portable fluorescence sensing, combined with advanced modelling methods to compensate readings for environmental interferences and false positives, can lay the foundations for a hybrid FIB sensing approach, allowing remote field deployment of a fleet of networked FIB sensors that can collect high-frequency data in near real-time. Such sensors will support proactive responses to sudden harmful faecal contamination events. A method is proposed to enable the development of the visioned FIB monitoring tool.

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Sensors for Monitoring Faecal Indicator Bacteria in Bathing Waters

Briciu-Burghina

Regan

2023

The Handbook of Environmental Chemistry

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Protocol for the recovery and detection of Escherichia coli in environmental water samples

Cited by 21 publications

References 34 publications

β-glucuronidase activity determination as an indirect estimate of Escherichia coli: development of a miniaturized assay and its application to seawater samples

β-glucuronidase activity determination as an indirect estimate of Escherichia coli: development of a miniaturized assay and its application to seawater samples

Monitoring Approaches for Faecal Indicator Bacteria in Water: Visioning a Remote Real-Time Sensor for E. coli and Enterococci

Sensors for Monitoring Faecal Indicator Bacteria in Bathing Waters

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