Surveillance and characterisation by pulsed-field gel electrophoresis of Cronobacter spp. in farming and domestic environments, food production animals and retail foods.

Cronobacter spp. (formally Enterobacter sakazakii) has been linked to illness in infants from contaminated powdered infant formula, however, there is limited information on the environmental sources and potential transmission routes of this pathogen. The aim of this study was to establish if food production animals (cattle, pigs), and the wider farm environment were playing a role in the transmission of Cronobacter spp. and also to assess the risk of cross contamination in the home where infant formula is prepared, from the presence of the pathogen on other foods and the general domestic environment. A wide range of samples (n=518) was collected at dairy farms, meat abattoirs, retail food stores and domestic environs and examined for the pathogen using an adapted ISO/DTS 22964 cultural protocol. The modified method included incubation at 42 degrees C instead of 44 degrees C and serial dilution of the enriched media prior to plating on Druggan-Forsythe-Iversen agar. Presumptive Cronobacter spp. colonies were confirmed by Real Time PCR targeting the dnaG on the MMS operon. All Cronobacter spp. isolated were speciated using biochemical tests, tested for resistance to 8 antibiotics and characterised using pulsed field gel electrophoresis. Cronobacter spp. was not recovered from cattle faeces, farm soil or trough water but isolates (n=33) were recovered from a variety of other sample types including cattle feed, pork and beef cuts, beef burgers and beef mince, green vegetables as well as organic breakfast cereals and domestic vacuum cleaner dust. The species recovered included C. Sakazakii (n=21), C. malonaticus (n=1) and C. turicensis (n=1). Of the 33 isolates 51% were resistant to Cephalothin but sensitive to all other 7 tested antibiotics. Sub-typing of the recovered isolates by PFGE showed considerable clonal diversity, though a number of persistent PFGE profiles were observed. In conclusion the study showed that Cronobacter spp. was not carried by food production animals but was present in a range of diverse sample types and environs with particular association with dry environments.