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      Re-examination of the taxonomic status of Enterobacter helveticus, Enterobacter pulveris and Enterobacter turicensis as members of the genus Cronobacter and their reclassification in the genera Franconibacter gen. nov. and Siccibacter gen. nov. as Franconibacter helveticus comb. nov., Franconibacter pulveris comb. nov. and Siccibacter turicensis comb. nov., respectively

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          Abstract

          Recently, a taxonomical re-evaluation of the genus Enterobacter , based on multi-locus sequence typing (MLST) analysis, has led to the proposal that the species Enterobacter pulveris , Enterobacter helveticus and Enterobacter turicensis should be reclassified as novel species of the genus Cronobacter . In the present work, new genome-scale analyses, including average nucleotide identity, genome-scale phylogeny and k-mer analysis, coupled with previously reported DNA–DNA hybridization values and biochemical characterization strongly indicate that these three species of the genus Enterobacter are not members of the genus Cronobacter , nor do they belong to the re-evaluated genus Enterobacter . Furthermore, data from this polyphasic study indicated that all three species constitute two new genera. We propose reclassifying Enterobacter pulveris and Enterobacter helveticus in the genus Franconibacter gen. nov. as Franconibacter pulveris comb. nov. (type strain 601/05 T = LMG 24057 T = DSM 19144 T) and Franconibacter helveticus comb. nov. (type strain 513/05 T = LMG 23732 T = DSM 18396 T), respectively, and Enterobacter turicensis in the genus Siccibacter gen. nov. as Siccibacter turicensis comb. nov. (type strain 508/05 T = LMG 23730 T = DSM 18397 T).

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          Genetic features of blaNDM-1-positive Enterobacteriaceae.

          Genetic features associated with the bla(NDM-1) gene were investigated in 6 Escherichia coli, 7 Klebsiella pneumoniae, 1 Citrobacter freundii, 1 Proteus mirabilis, and 1 Providencia stuartii isolate of worldwide origin. Clonal diversity was observed for both E. coli and K. pneumoniae. The bla(NDM-1) gene was carried by different plasmid types (IncA/C, IncF, IncL/M, or untypeable) and was likely chromosome borne in two isolates. The bla(NDM-1) plasmids coharbored a variety of resistance determinants, including β-lactamase genes, quinolone resistance genes, and 16S RNA methylase genes.
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            Cronobacter gen. nov., a new genus to accommodate the biogroups of Enterobacter sakazakii, and proposal of Cronobacter sakazakii gen. nov., comb. nov., Cronobacter malonaticus sp. nov., Cronobacter turicensis sp. nov., Cronobacter muytjensii sp. nov., Cronobacter dublinensis sp. nov., Cronobacter genomospecies 1, and of three subspecies, Cronobacter dublinensis subsp. dublinensis subsp. nov., Cronobacter dublinensis subsp. lausannensis subsp. nov. and Cronobacter dublinensis subsp. lactaridi subsp. nov.

            [Enterobacter] sakazakii is an opportunistic pathogen that can cause infections in neonates. This study further clarifies the taxonomy of isolates described as [E.] sakazakii and completes the formal description of the proposed reclassification of these organisms as novel species and subspecies within a proposed novel genus, Cronobacter gen. nov. [E.] sakazakii was first defined in 1980, however recent polyphasic taxonomic analysis has determined that this group of organisms consists of several genomospecies. In this study, the phenotypic descriptions of the proposed novel species are expanded using Biotype 100 and Biolog Phenotype MicroArray data. Further DNA-DNA hybridization experiments showed that malonate-positive strains within the [E.] sakazakii genomospecies represent a distinct species, not a subspecies. DNA-DNA hybridizations also determined that phenotypically different strains within the proposed species, Cronobacter dublinensis sp. nov., belong to the same species and can be considered as novel subspecies. Based on these analyses, the following alternative classifications are proposed: Cronobacter sakazakii gen. nov., comb. nov. [type strain ATCC 29544(T) (=NCTC 11467(T))]; Cronobacter malonaticus sp. nov. [type strain CDC 1058-77(T) (=LMG 23826(T)=DSM 18702(T))]; Cronobacter turicensis sp. nov. [type strain z3032(T) (=LMG 23827(T)=DSM 18703(T))]; Cronobacter muytjensii sp. nov. [type strain ATCC 51329(T) (=CIP 103581(T))]; Cronobacter dublinensis sp. nov. [type strain DES187(T) (=LMG 23823(T)=DSM 18705(T))]; Cronobacter dublinensis subsp. dublinensis subsp. nov. [type strain DES187(T) (=LMG 23823(T)=DSM 18705(T))]; Cronobacter dublinensis subsp. lausannensis subsp. nov. [type strain E515(T) (=LMG 23824=DSM 18706(T))], and Cronobacter dublinensis subsp. lactaridi subsp. nov. [type strain E464(T) (=LMG 23825(T)=DSM 18707(T))].
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              Cronobacter condimenti sp. nov., isolated from spiced meat, and Cronobacter universalis sp. nov., a species designation for Cronobacter sp. genomospecies 1, recovered from a leg infection, water and food ingredients.

              A re-evaluation of the taxonomic position of five strains, one assigned to Cronobacter sakazakii (strain 1330(T), isolated from spiced meat purchased in Slovakia), two previously assigned to Cronobacter genomospecies 1 (strains NCTC 9529(T) and 731, isolated from water and a leg infection, respectively) and two previously assigned to Cronobacter turicensis (strains 96 and 1435, isolated from onion powder and rye flour, respectively) was carried out. The analysis included phenotypic characterization, 16S rRNA gene sequencing and multilocus sequence analysis (MLSA) of seven housekeeping genes (atpD, fusA, glnS, gltB, gyrB, infB, ppsA; 3036 bp). 16S rRNA gene sequence analysis and MLSA showed that strain 1330(T) formed an independent phylogenetic lineage in the MLSA, with Cronobacter dublinensis LMG 23823(T) as the closest neighbour. DNA-DNA reassociation and phenotypic analysis revealed that strain 1330(T) represented a novel species, for which the name Cronobacter condimenti sp. nov. is proposed (type strain 1330(T) = CECT 7863(T) = LMG 26250(T)). Strains NCTC 9529(T), 731, 96 and 1435 clustered together within an independent phylogenetic lineage, with C. turicensis LMG 23827(T) as the closest neighbour in the MLSA. DNA-DNA reassociation and phenotypic analysis confirmed that these strains represent a novel species, for which the name Cronobacter universalis sp. nov. is proposed (type strain NCTC 9529(T) = CECT 7864(T) = LMG 26249(T)).
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                Author and article information

                Journal
                Int J Syst Evol Microbiol
                Int. J. Syst. Evol. Microbiol
                IJSEM
                ijs
                International Journal of Systematic and Evolutionary Microbiology
                Society for General Microbiology
                1466-5026
                1466-5034
                October 2014
                October 2014
                : 64
                : Pt 10
                : 3402-3410
                Affiliations
                [1 ]Institute for Food Safety and Hygiene, University of Zurich, Zurich, Switzerland
                [2 ]CFSAN, FDA, Laurel, USA
                [3 ]UCD Centre for Food Safety, School of Public Health, Physiotherapy & Population Science, University College, Dublin, Ireland
                [4 ]WHO Collaborating Centre for Cronobacter, Belfield, Dublin, Ireland
                Author notes
                Correspondence Roger Stephan stephanr@ 123456fsafety.uzh.ch

                The GenBank/EMBL/DDBJ accession numbers for the whole-genome shotgun projects of Enterobacter turicensis 508/05 T ( = LMG 23730 T = DSM 18397 T) and 610/05 ( = LMG 23731) are BioProject Accession: PRJNA215839, Biosample numbers: SAMN02319256 and SAMN02319257, accession numbers: AVPP00000000 and AXDM00000000, respectively; those of Enterobacter pulveris 601/05 T ( = LMG 24057 T = DSM 19144 T) and 1160/04 ( = LMG 24058 = DSM 19146) are BioProject Accession: PRJNA222260, Biosample numbers: SAMN02369274 and SAMN02369275, accession numbers: AXSY00000000 and AXSZ00000000, respectively; and those of Enterobacter helveticus 513/05 T ( = LMG 23732 T = DSM 18396 T) and 1159/04 ( = LMG 23733) are BioProject Accession: PRJNA 221525, Biosample numbers: SAMN02364109 and SAMN02364110, accession numbers AXDK00000000 and AXDL00000000, respectively.

                Article
                059832
                10.1099/ijs.0.059832-0
                4179279
                25028159
                479fd928-1281-4b67-8766-399a3f04e96a

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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                New Taxa
                Proteobacteria
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                Microbiology & Virology
                Microbiology & Virology

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