Growth kinetics and hydrolytic enzyme production of Pseudomonas spp. isolated from pasteurized milk

Psychrotrophs, particularly Pseudomonas spp. are known to be the main determinants of the shelf-life of pasteurized milk and refrigerated raw milk. It is presumed that they mainly cause spoilage through the elaboration of proteinase and lipase enzymes. At the time of this research, under the relevant European Directive, one of the means of determining the quality of pasteurized milk was the pre-incubated count, which involves incubating the milk sample for 5 d at 6 degrees C followed by a plate count. Examination of numerous pre-incubated counts revealed a bimodal rather than a normal distribution indicating that the types of contaminants in pasteurized milk may be as important as their initial concentration. Pseudomonads that gave particularly high (> 5 x 10(6) cfu/ml) and low (< 10(3) cfu/ml) pre-incubated counts were isolated (high and low count isolates respectively). After the organisms had been subjected to a cold shock no consistent trend between the groups of isolates was detected with respect to lag phase duration. However, the high count isolates consistently had a faster exponential growth rate. Unexpectedly, with the exception of one isolate, the low count isolates produced detectable proteinase and lipase earlier. In addition, with one exception, maximal proteinase and lipase production was observed with the low count isolates. These findings indicate that there is no causal relationship between selective growth advantage and ability to produce proteinase and lipase. It also indicates that the spoilage of pasteurized milk is a complex phenomenon and is worthy of further research.