Aim: Enterobacter cloacae complex bacteria are of both clinical and environmental importance. Phenotypic methods are unable to distinguish between some of the species in this complex, which often renders their identification incomplete. The goal of this study was to develop molecular assays to identify Enterobacter hormaechei and Ent. cloacae genetic cluster III which are relatively frequently encountered in clinical material. Methods and Results: The molecular assays developed in this study are qPCR technology based and served to identify both Ent. hormaechei and Ent. cloacae genetic cluster III. qPCR results were compared to hsp60 sequence analysis. Most clinical isolates were assigned to Ent. hormaechei subsp. steigerwaltii and Ent. cloacae genetic cluster III. The latter was proportionately more frequently isolated from bloodstream infections than from other material (P < 0Á05). Conclusion: The qPCR assays detecting Ent. hormaechei and Ent. cloacae genetic cluster III demonstrated high sensitivity and specificity. Significance and Impact of the Study: The presented qPCR assays allow accurate and rapid identification of clinical isolates of the Ent. cloacae complex. The improved identifications obtained can specifically assist analysis of Ent. hormaechei and Ent. cloacae genetic cluster III in nosocomial outbreaks and can promote rapid environmental monitoring. An association was observed between Ent. cloacae cluster III and systemic infection that deserves further attention.
S. faecium predominated in fresh vegetables which were not irrigated with sewage treatment plant water. Vegetables which were irrigated with waste water from a sewage treatment plant contained equal numbers of the two species. S. faecium survived the effect of chlorination better than S. faecalis. The tendency for chain aggregate formation was more prominent in S. faecalis especially during exponential growth. This affected the recoveries after freezing but not after the chlorination treatment. With exponential cultures which were treated for aggregate dispersal and with stationary cultures, S. faecium survived the effect of freezing better. This indicates a higher resistance of this organism and that the vegetable treatments in the freezing plant may result in a relative enrichment in S. faecium. Thus, enterococcal count in frozen vegetables is comprised to a large extent of this group, a fact which reduces the significance of the enterococci as a sanitary indicator. A specific S. faecalis count may be a better indicator of the sanitary quality of frozen vegetables and may be performed with KF agar supplemented with 0.04% K2 Te03.
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