Rapid and miniaturized method for detection of hygiene indicators,Escherichia coli and coliforms,in dairy products

Current investigation was aimed to develop colorimetric tests for rapid detection of Escherichia coli/coliforms. These test (s) for E. coli and coliforms were developed using the modified E. coli selective medium (M-ECSM) and coliform selective medium, respectively. The selective media contain a combination of group specific marker enzymes and selective agents. The marker enzymes were screened using chromogenic substrates wherein β-D-glucuronidase and glutamate decarboxylase were found specific for E. coli while β-D-galactosidase for coliforms. The selectivity of the media was achieved using different concentrations of ampicillin and gentamicin. The optimized test procedures enabled sensitive detection of 0.35 ± 0.10 log cfu/ml of E. coli and 0.57 ± 0.15 log cfu/ml of coliforms at 37°C within 14.30 ± 0.45 and 12.15 ± 0.30 hr, respectively. M-ECSM selectively inhibited major Enterobacteriaceae contaminants (Salmonella, Shigella, and Yersinia) up to 6 log cfu/ml. Moreover, better selectivity of M-ECSM was reported against tested commercial chromogenic media. Field evaluation of developed test (s) reported prevalence of E. coli/coliforms as 57.29/88.54% in 96 raw milk and 16.28/51.16% in 43 pasteurized milk samples. Further, test components were vacuum dried in the form of miniaturized point-of-need test for field application in dairy farms and industries with minimal infrastructural requirements.