This study applied the Cronobacter spp. multilocus sequence typing (MLST) scheme to three strain collections, then known as Enterobacter sakazakii, which had been isolated between 1988 and 2009 from 14 countries. The results revealed the predominance (85%) of C. sakazakii (72 strains) in all three collections. The remaining strains were C. turicensis (10%), C. malonaticus (4%), and C. muytjensii (1%). No strains of C. dublinensis, C. universalis or C. condimenti were identified. Twenty-one out of seventy two C. sakazakii strains were in the clinically significant ST4 clonal complex, and were found in all three strain collections. These results confirm C. sakazakii ST4 is one of the predominant clonal complexes over the past 20 years in several parts of the world. Further understanding of the ecosystem and sources of the organism may be used for the development of improved intervention strategies in the diary industry.

Aims:  To characterise the occurrence of Cronobacter in milk powder factories. Methods and Results:  Cronobacter was isolated from 32% of 298 environmental samples from five factories. More isolations occurred in nonprocessing (49%) than processing areas (29%), although the greatest occurrence was in a single milk powder area during shutdown maintenance (81%) and the lowest after reinstatement of production hygiene practices (6%). Clonal analysis using PFGE placed 129 isolates into 49 groups. Most clones (45) were unique to each factory and seven were isolated in both milk powder and other areas of the same factory including tanker bays, evaporator rooms, an employee’s shoes and external roofs. Cronobacter was not isolated from raw milk processing areas. Within powder areas, 17 clones occurred at more than one and up to eight locations and six occurred more than once at the same location. Between four and seven clones were in the powder areas at each factory. The most prevalent and persistent clones were isolated from external roofs above spray driers, in air treatment areas and where high foot traffic occurs. Conclusions:  Cronobacter is dispersed widely at milk powder factories. This study suggests that distribution is assisted by movement of air, milk powder and personnel and that new hygiene strategies will be needed to reduce prevalence. Significance and Impact of the Study:  Knowledge of occurrence is essential for the development of strategies to control dissemination of Cronobacter within factories and reduce risk of entry into powdered milk products.

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