Suitability of total coliform beta-D-galactosidase activity and CFU counts in monitoring faecal contamination of environmental water samples
- Authors: Wutor, V C , Togo, C A , Pletschke, Brett I
- Date: 2009
- Language: English
- Type: Article
- Identifier: vital:6477 , http://hdl.handle.net/10962/d1006165 , http://www.scielo.org.za/scielo.php?pid=S1816-79502009000100010&script=sci_arttext
- Description: Total coliforms are a group of bacteria found in high numbers in mammalian intestines; hence their presence in water indicates the possible contamination with faecal material. Total and faecal coliform counts were monitored over a period of 18 months using mFC, m-Endo and CM1046 media together with enzymatic assays on 215 environmental water samples obtained from the Eastern Cape Province of South Africa. A positive correlation, with an R2 value of 0.9393 was observed between faecal and total coliform colony units employing mFc and m-Endo media, and 0.8818 using CM1046 media. Also, a positive correlation was observed between Escherichia coli colony-forming units and β-D-galactosidase (B-GAL) activity (R2=0.8542). Overall, this study indicated that faecal contamination of environmental water samples could be monitored by measuring total coliform β-galactosidase activity and total coliform colony-forming units.
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- Date Issued: 2009
A novel biosensor for the detection and monitoring of -d-galactosidase of faecal origin in water
- Authors: Wutor, V C , Togo, C A , Limson, Janice L , Pletschke, Brett I
- Date: 2007
- Language: English
- Type: Article
- Identifier: vital:6468 , http://hdl.handle.net/10962/d1005797 , http://dx.doi.org/10.1016/j.enzmictec.2006.10.039
- Description: A voltammetric sensor prepared by the immobilization of metallophthalocyanine complexes onto a glassy carbon electrode has been developed for the detection of β-d-galactosidase (B-GAL) of faecal origin in water. Electrooxidation of chlorophenol red, a breakdown product of the chromogenic substrate chlorophenol red β-d-galactopyranoside, was used as a measure of β-d-galactosidase activity. At metallophthalocyanine modified electrodes, in particular copper(II) phthalocyanine, a decrease in electrode fouling was observed. The sensor was sensitive to fluctuations in pH, not significantly affected by temperature variations and could detect one colony forming unit/100 mL in 15 min. Loss of 40% sensitivity was observed over a period of 30 days. A strong correlation between sensor sensitivity and colony forming units was observed. The sensor is capable of detecting viable but nonculturable bacteria, overcoming this drawback of the use of culture media for detection of coliforms.
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- Date Issued: 2007
Comparison of the direct enzyme assay method with the membrane filtration technique in the quantification and monitoring of microbial indicator organisms - seasonal variations in the activities of coliforms and E.coli, temperature and pH
- Authors: Wutor, V C , Togo, C A , Pletschke, Brett I
- Date: 2007
- Language: English
- Type: Article
- Identifier: vital:6479 , http://hdl.handle.net/10962/d1006241
- Description: The aim of this project was to monitor variations and relationships between coliform and E. coli counts, the activities of their marker enzymes GAL and GUD, and temperature and pH over a period of 12 months in river samples obtained from the Eastern Cape, South Africa. Several polluted water samples were collected for direct coliform β-D-galactosidase (B-GAL) and Escherichia coli β-D-glucuronidase (B-GUD) assays and the membrane filtration technique. While all the samples showed enzyme activities, not all exhibited growth on CM1046 media. Variation in B-GAL activity (40%) was observed between November (highest activity month) and May (lowest activity month). The highest and lowest B-GUD activities were observed in the months of September and May/June, respectively. The sensitivity of the spectrophotometric assay method was indicated by a limit of detection (LOD) of 1 coliform forming unit (CFU)/100 mℓ and 2 CFU/100 mℓ for coliforms and E. coli, respectively. There was a significant (P < 0.05) positive correlation between E. coli counts and GUD activity (R2 = 0.8909). A correlation of R2 = 0.9151 was also observed between total coliforms and B-GAL activity, even though the CFUs were not evenly distributed. Direct enzyme assays were also shown to be more sensitive than the membrane filtration (MF) technique.
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- Date Issued: 2007
The effect of physico-chemical parameters and chemical compounds on the activity of β-d-galactosidase (B-GAL), a marker enzyme for indicator microorganisms in water
- Authors: Wutor, V C , Togo, C A
- Date: 2007
- Language: English
- Type: text , Article
- Identifier: vital:6470 , http://hdl.handle.net/10962/d1005800 , https://doi.org/10.1016/j.chemosphere.2007.02.050
- Description: The presence of coliforms in polluted water was determined enzymatically (in situ) by directly monitoring the activity of beta-d-galactosidase (B-GAL) through the hydrolysis of the yellow chromogenic subtrate, chlorophenol red beta-d-galactopyranoside (CPRG), which produced a red chlorophenol red (CPR) product. The objectives of this study were to monitor the effect of compounds commonly found in the environment and used in water treatment on a B-GAL CPRG assay and to investigate the differences between the environmental B-GAL enzyme and the pure commercial enzyme. Environmental B-GAL was optimally active at pH 7.8. Two temperature optima were observed at 35 and 55 degrees C, respectively. B-GAL activity was strongly inhibited by silver and copper ions. While calcium and ferrous ions at lower concentrations (50-100mgl(-1)) increased the enzyme activity, a reduction was observed at higher concentrations (200mgl(-1)). Sodium hypochlorite, normally used in rural areas to disinfect water gradually decreased B-GAL activity at concentrations between 0 and 5600ppm for both the commercial and environmental enzymes. B-GAL from the environment behaved differently from its commercially available counterpart.
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- Date Issued: 2007
The effect of sulfide on -glucosidases: implications for starch degradation in anaerobic bioreactors
- Authors: Wutor, V C , Togo, C A
- Date: 2007
- Language: English
- Type: Article
- Identifier: vital:6463 , http://hdl.handle.net/10962/d1005792 , dx.doi.org/10.1016/j.chemosphere.2006.03.011
- Description: Membrane associated α-glucosidase activity was investigated in a methanogenic bioreactor (MR) and a biosulfidogenic bioreactor (SR). Temperature and pH optima studies showed temperature optima of 50 °C and pH optima of 8.0 for the α-glucosidases from both the MR and SR. Sulfide (at a concentration of 150 mg l[superscript (−1)]) resulted in the complete loss of all α-glucosidase activity in both the MR and SR. β-Glucosidase activities in our bioreactors were previously shown to be stimulated in the presence of sulfide. α-Glucosidases, in contrast, are inhibited by sulfide. This differential effect of sulfide on α-glucosidase and β-glucosidase activities is highlighted and is of crucial consequence to the respective degradation and utilization of starch and cellulose substrates in natural anaerobic environments and anaerobic bioreactors specifically designed for the accelerated digestion of wastewater sludge under biosulfidogenic conditions.
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- Date Issued: 2007