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Development of an enzyme assay for rapid assessment of Escherichia coli in seawaters (Articolo in rivista)

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Development of an enzyme assay for rapid assessment of Escherichia coli in seawaters (Articolo in rivista) (literal)

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G. Caruso, Crisafi E., Mancuso M (2002)
Development of an enzyme assay for rapid assessment of Escherichia coli in seawaters
in Journal of applied microbiology (Print); Blackwell, Oxford (Regno Unito)
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prodotto con 19 citazioni SCOPUS, 14 citazioni ISI, 30 citazioni Google Scholar, SJR 2011 : 0.864 - SJR 2009: 0.812, IF 2009: 2.098 (literal)

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The paper concerns the development of rapid methods for the determination of Escherichia coli, through the of an experimental assay using a fluorogenic substrate for b-glucuronidase (MUG test). Since this enzyme is specific of E.coli, the MUG assay is proposed as an indirect way to evaluate the presence of this indicator organism in seawaters. The procedure represents one of the analytical protocols developed at the Institute for Marine Coastal Environment Section of Messina to deal with the increasing request of new microbiological methods for environmental monitoring. Its reliability for natural samples has been tested in comparison with the fluorescent antibody and culture methods. The statistical analysis through linear regression of data obtained by the three methods pointed out a significant relationship between the enzyme and microscopic data, indicating a greater specificity of the assay for E.coli rather than for the whole coliform group. The statistical relationship between enzyme values and culture counts was higher for heavily polluted samples, containing at least or more than 100 coliforms per 100ml; in these conditions, the high organic matter available sustains the presence of metabolically active cells. (literal)

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Titolo

Development of an enzyme assay for rapid assessment of Escherichia coli in seawaters (literal)

Abstract

Aims: An analytical protocol has been developed and applied for the detection of glucuronidase activity in marine waters as a rapid alternative approach to assess the microbiological quality of seawaters. Methods and Results: The fluorogenic substrate 4-methylumbelliferyl-?-D-glucuronide is cleaved to a fluorescent product, methylumbelliferone, by the enzyme ?-glucuronidase, specific to Escherichia coli and closely related enterobacterial species (Shigella). The results suggest that this test is related to E. coli numbers, as estimated by immunofluorescence, more significantly than to faecal coliform numbers, obtained from culture media. Conclusions: The determination of the potential rate of glucuronidase activity may be used as a diagnostic tool for the indirect estimation of the presence of E. coli in seawaters. Significance and Impact of the Study: The method may be particularly useful in the early warning of seawater pollution, allowing the screening of coastal areas with different contamination levels in reduced time. (literal)

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