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Practical use and development of biomérieux TEMPO® system in microbial food safetyAlsaadi, Yousef Saeed January 1900 (has links)
Doctor of Philosophy / Department of Food Science / Daniel Y.C. Fung / In the food industry, coliform testing is traditionally done by the time consuming and labor intensive plate count method or tube enumeration methods. The TEMPO® system (bioMérieux, Inc.) was developed to improve laboratory efficiency and to replace traditional methods. It uses a miniaturization of the Most Probable Number (MPN) method with 16 tubes with 3 dilutions in one single disposable card. It utilizes two stations: the TEMPO® Preparation station and the TEMPO® Reading station. In this study, the Oxyase® (Oxyase®, Inc.) enzyme was added to TEMPO® CC (Coliforms Count), TEMPO® AC (aerobic colony count) and TEMPO® EC (E. coli Count) methods. Water samples of 1 ml with 0.1 ml of Oxyase® enzyme were compared to samples without the Oxyase® enzyme using the TEMPO® system. Samples were spiked with different levels of coliforms (10, 102, 103 and 104 CFU/ml), stomached (20 sec), and pipetted into the three different TEMPO® media reagents (4 ml) in duplicate and then automatically transferred into the corresponding TEMPO® cards by the TEMPO® preparation station. Counts were obtained using the TEMPO® reading station after 8, 12, 16, 22 and 24 hours at an incubation temperature of 35°C. Results from 20 replicates were compared statistically. Using TEMPO® tests, high counts in food samples (>6 log 10 CFU/ml) can be read in 6±2 hours of incubation using the time-to-detection calibration curve. The TEMPO® system reduces reading time (reading protocol should be changed). There is no need to wait for 22 hours of incubation only 12 hours is required. Oxyrase® enzyme is not needed for the TEMPO® system.
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