Investigation of the Use of a Cocktail of Lux-Tagged Cronobacter Strains for Monitoring Growth in Infant Milk Formulae

Abstract

The objective of the present study was to create a collection of Lux-tagged Cronobacter strains to determine whether bioluminescence could be used to monitor growth of this pathogen in infant milk formula (IMF). Nine Cronobacter strains (seven C. sakazakii, one C. malonaticus, and one C. muytjensii) were transformed with plasmid p16S lux, and integration of the plasmid at the desired site on the chromosome was confirmed by PCR. The integrated plasmid was stable in the absence of antibiotic selection, and growth of the Lux-tagged strains was similar to that of their nontagged counterparts. Growth of Lux-tagged strains was monitored in real time in 10 commercial brands of IMF by measuring light emission with a luminometer. Although all of the IMF samples tested were able to support the growth of the Cronobacter strains, differences were observed among IMF brands. Variations in the amount of light emitted by individual Cronobacter strains were also noted. Monitoring light emission with a combination of two strains that produced higher and lower than average relative light readings was a good surrogate for evaluating the entire collection of Lux-tagged strains.

Development of Bioluminescent Cronobacter sakazakii ATCC 29544 in a Mouse Model

“…This bioluminescence tool has recently been used to monitor C. sakazakii in infant milk formula (6,14). However, no good method for real-time monitoring of C. sakazakii infection in animals has been developed.…”