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Alkaline Phosphatase Enzyme Activity Measurement as a robust on-line Monitoring Assay for Water Quality Assessment

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Alkaline Phosphatase Enzyme Activity Measurement as a robust on-line Monitoring Assay for Water Quality Assessment

Rechten: Alle rechten voorbehouden

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The analysis of contaminant concerns a number of organizations, including industrial sectors (e.g. agrifood, water, and healthcare), regulatory authorities and food control and environmental agencies. (Bio)sensors form important analytical tools, as they are comparable to or better than the conventional methods. Conventional methods of detection are very sensitive but equally they are too laborious, time-consuming and mostly depended on the properties of bacteria to form frequent colonies. Additionally, these methods do not allow for easily continuous monitoring because they need well-trained operators and in most cases they require steps of extractions and sample preparation or pretreatment.
Biosensors offered alternative assay formats as they are able to fulfill some demands that the classic methods of analysis do not attain. In general, the analytical devices composed of biological recognition elements (e.g. enzyme, antibody, receptor or microorganism) coupled to physical or chemical transducer (mass, optical, thermal and electrochemical). Biosensors offers, sensitivity, selectivity, specificity, reliability, and robustness. Some other advantages, they are rapid, on-site, and mostly have simple and portable equipment construction.
Biosensors that use enzymes as the bio-recognition elements are widely developed and well-studied area. Here, the turnover number of the enzyme to its capture substrate, provide a significant amplification system with a high degree of specificity and sensitivity detection level.
Alkaline Phosphatase (ALP), is an enzyme which catalyzes the hydrolysis of various monophosphate esters at alkaline pH (>9). ALP is presented in many tissues of all living beings from bacteria to mammals. The enzyme with its wide specificity and activity has been used in biosensors for the detection of different contaminates. BACTcontrol is one of the three different devices that produced by microLAN company – Waalwijk. BACTcontrol measures microbial ALP activity using a high-sensitivity fluorescent substrate that, upon binding with the enzyme produce strong fluorescence signal that is monitored permanently.
The aim of this study that was carried out by microLAN Company in Waalwijk – Netherlands, is to validate BACTcontrol results and to assess its efficiency as a useful method for changes in the biological parameter of different water types. The ALP turnover rate measured from BACTcontrol was compared with different approved methods that routinely used to assess drinking water quality. The data were compared with Flow Cytometer total and intact cells count (FCM), Microscopic Total Cells Count (TCC), Microscopic ELF-labeled cells count for microbial cells with alkaline phosphatase activity,
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total and cell bounded Adenosine Tir-Phosphate measurement (ATP) and the conventional Heterotrophic Plate Count (HPC).
The study showed that the enzymatic assay using BACTcontrol correlated well with most of the approved methods tested. Within similar water types, BACTcontrol correlated well with the flow cytometric total and intact cells counts , total and cell bounded ATP, ELF-microscopic cells count and with microscopic intact cells count using viable stains, (R2= 0.99, 0.89, 0.83 and 0.92 respectively). There was considerable loss of correlation after data from different water types compared together (e.g. tap water and natural surface water ). In natural fresh water, ALP activity measured with BACTcontrol was prone to interference from the extracellular cell-free substances.
No correlation was observed between BACTcontrol and the traditional Heterotrophic plate count (R2 = -0.32). The microscopic and flow cytometric bacterial counts paralleled but microscopic counts always gave lower counts than FCM (<21%).
These results support the use of BACTcontrol as a rapid, reproducible and relatively low-cost tool for routine monitoring purposes of water quality, it can be used to trigger alarms when significant changes are seen in the environment, but further laboratory investigations are needed to assess water safety or to detect specific target bacteria.

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Trefwoorden
OrganisatieAvans Hogeschool
OpleidingBiologie en Medisch Laboratoriumonderzoek-Breda
AfdelingATGM Academie voor de technologie van Gezondheid en Milieu
PartnersmicroLAN
Datum2016-08-01
TypeBachelor
TaalEngels

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