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A rapid and selective method for monitoring the growth of coliforms in milk using the combination of amperometric sensor and reducing of methylene blue
Authors:Yuan-Geuy  Hung-Yi  Chuan-Liang  Hong-Jen  Chiun-Jye  Hung-Der  
Affiliation:aDepartment of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 912, Taiwan;bDepartment of Biological Science and Technology, National Chiao Tung University, 75 Po-Ai Street, Hsinchu 300, Taiwan;cDepartment of Food Science, Tunghai University, No. 181, Section 3, Taichung Harbor Road, Taichung 407, Taiwan;dDepartment of Food Science, Yuanpei University, No. 306, Yuanpei Street, Hsinchu 300, Taiwan
Abstract:A novel method for monitoring the growth of coliforms in milk was developed based on measuring the current change in an amperometric sensor. The sensor consists of a circuit with a homemade potentiostat and a pair of electrodes. The electrode was immersed in milk samples containing methylene blue with various concentrations of bacterial inoculums. The microbial metabolism led to the reduction of methylene blue resulting in a change of current. The time required to identify readily detectable change (detection time, DT) provided an approximate measurement of the amount of microorganisms in the initial inoculums. The sensor system used in this study has the selectivity towards coliform bacteria such as Escherichia coli and Enterobacter aerogenes. The calibration curve of DT against concentration of coliform showed a linear correlation coefficient (R2 = 0.9192) over the range of 102–108 CFU/mL. The sensor was able to detect the coliform bacteria at initial concentrations of 105 CFU/mL within 6 h, making it suitable for use in real-time monitoring of bacterial growth. This system has potential application in the detection of coliform concentration in milk at dairy farms when a proper selective media is designed.
Keywords:Amperometric sensor  Methylene blue  Coliforms  Detection time
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