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Application of a membrane filtration method for the detection of gram-negative bacterial contamination in pasteurized milk

Authors: W.S. Kwee, T.W. Dommett and A.C. Vos


A method involving membrane filtration followed by culturing on selective agar was developed to improve the sensitivity of detecting Gram negative bacteria in pasteurized milk. The method involves enzymatic pretreatment of a milk sample at 43 to 49°C for 10 min and rapid filtration of 10 to 14 mL through a 47 mm diameter, 0.6mm pore size polycarbonate membrane filter, with minimal loss in apparent total number of bacteria recovered. A Gram negative selective medium (PBAC-ANS) allowed the growth of 98% of 50 Gram negative isolates and prevented the growth of 92.4% of 79 Gram positive isolates. Of the isolates which grew on the medium, 71.4% of the Gram negatives showed flurescent intensity of ≥ 2 and 83.5% of the Gram positives had fluorescent intensity of < 2.

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