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The use of an automated growth analyser to measure recovery times of single heat-injured Salmonella cells
Authors:PJ Stephens  JA Joynson  KW Davies  R Holbrook  HM Lappin-Scott  & TJ Humphrey
Institution:Unilever Research, Colworth Laboratory, Sharnbrook, Bedford,;Department of Biological Sciences, Exeter University, Exeter, UK,;PHLS Food Microbiology Research Unit, Church Lane, Heavitree, Exeter, UK
Abstract:A new approach to the study of recovery times of single heat-injured Salmonella cells isdescribed. It comprises the generation of a standard heat-injured culture, serial dilution of thisculture to near extinction, inoculation of the serial dilutions across many microtitre plates andmeasurement of the subsequent recovery and growth using an automated turbidometric analyser.Lag times for individual cells were estimated from turbidity data using a model that accuratelyextrapolated the growth curve back to the starting inoculum level. Lag times were comparedusing a number of different commercially available pre-enrichment media. The most typical resultwas a very broad distribution of lag times at the single cell inoculum level, with many values inexcess of 20 h. Even at an inoculum level 10-fold higher, lag times for some injured cells wereestimated to be >10 h. More significantly, it was found that some media recovered more injuredcells than others and vice versa. Between the worst and best media there were as many as 3 log10 cycles difference in the number of cells recoverable. No trends were apparent linkingchoice of medium with performance. The implications of these findings, in relation to traditionaland rapid methodology, are discussed.
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