Rapid and Simple Detection of Coliform Bacteria in Milk Using Menadione-Catalyzed Luminol Chemiluminescence Assay

Abstract

Menadione-catalyzed luminol chemiluminescence assay was evaluated for rapid detection of coliform bacteria in milk. The assay was evaluated using raw milk (non-pasteurized) samples and pasteurized milk samples spiked with Escherichia coli, Enterobacter aerogenes, Klebsiella oxytoca, Citrobacter freundii. The detection limit in this assay was +* / ῌ+*. CFU/ml using a 30-well plate with BGLB broth. Our study revealed that coliform contamination of +* + ῌ+* * CFU/ml of milk could be detected after +2 h of incubation at-/ in BGLB medium. Moreover, this assay detected gas production in samples of BGLB after only +2 h of a .2-h incubation. As this assay is convenient, quick and inexpensive compared to the conventional method, it should be useful for the rapid detection of coliform contamination in pasteurized milk.

Selective colorimetric detection of Gram-negative re-contaminants in pasteurised milk products by a novel application of the BacT/ALERT 3D system

“…in pasteurised milk products. Compared with other published methods for the detection of re-contaminants in pasteurised milk products (Firstenberg-Eden et al, 2002;Kawasaki et al, 2007;Nicolaou & Goodacre, 2008;Ren & Hsieh, 2005;Soejima et al, 2012a), this method covers a broader range of re-contaminants within acceptable time periods. In addition, the sub-cultivation of isolates for further pheno-and genotypic analysis is possible.…”

“…Different microbiological parameters have been analysed using novel cultural techniques in pasteurised milk: coliforms (Foschino, Colombo, Crepaldi, & Baldi, 2003;Kang & Gray, 2002;Madden & Gilmour, 1995;Raybaudi, Zea, Curini, & Martínez, 2005), Pseudomonas species (Van Tassell et al, 2012), Escherichia coli and coliforms (Beloti, Barros, Nunes, De Santana, & Nero, 2002;Feldsine et al, 2005), total bacterial counts and coliforms (Ginn, Packard, & Fox, 1986), standard plate counts, psychrotrophic bacterial counts, and coliforms (Senyk, Kozlowski, Noar, Shipe, & Bandler, 1987) and total viable counts, coliforms, and Enterobacteriaceae (Firstenberg-Eden, Foti, McDougal, & Baker, 2002). Furthermore, novel detection techniques and procedures have been presented for the detection of recontaminants in pasteurised milk, such as the determination of microbial activity using a microrespirometer (Ren & Hsieh, 2005), the detection of coliforms by menadione-catalysed luminol chemiluminescence (Kawasaki et al, 2007), the detection of microbial spoilage of milk using Fourier transform infrared spectroscopy and chemometrics (Nicolaou & Goodacre, 2008), the application of a matrix-assisted laser desorption/ionisation timeof-flight mass spectrometry (MALDI-TOF-MS) and multivariate analysis (Nicolaou, Xu, & Goodacre, 2012), and finally the specific detection of total viable coliforms, as well as viable Enterobacteriaceae, by PCR-based techniques (Soejima, Minami, Ayeshima, & Iwatsuki, 2012a;Soejima, Minami, Yaeshima, & Iwatsuki, 2012b). However, these methods are neither capable of detecting very low numbers of re-contaminants nor do they cover the spectrum of microbes, which may play some role under practical conditions.…”

Rapid Enumeration of Microbial Cells in Solubilized Soymilk Using an Automatic Cell Counting System with LED Illumination

“…Therefore, a simple, rapid, and accurate detection method is desirable. Several non-culture methods have been used for the rapid detection of bacteria in food (Fujikawa and Morozuki, 2003;Gracias and Mckillip, 2004;Kawasaki et al, 2004;Kawasaki et al, 2007;Masuda-Nishimura et al, 2000;Miyamoto et al, 1998). PCR is a useful technique because of its high sensitivity.…”