Nine different strains, CTCBSIT to CTCBS9, were identified to be the causative agents of black spots on the surface of raw cured meat products. The formation of black spots under aerobic conditions is reproducible upon reinoculation of meat products with any of these strains, indicating that they are the causative agent. The strains were Gram-negative, catalase-positive and obligately aerobic rods. The G+C content of DNA of strain CTCBSIT is 56.0203 mol%. The content of non-polar main fatty acids were 16:0,16:1,18:1 and 19:0 cyc. Its phylogenetic position was elucidated by comparative sequence analysis of the 165 rRNA gene. Overall sequence similarity to other bacteria does not exceed 93.3 %. Isolate CTCBSIT clustered phylogenetically within the y-subclass of the Proteobacteria and is closely related to members of Halomonas (90.5-91.9 %) and to Zymobacter palmae (93-3 %). A genetic homogeneity of the nine strains was demonstrated by M13 random amplified polymorphic DNA-PCR, whereas differentiation from other genera, e.g. Zymobacter and Pseudomonas, could easily be achieved by their chemotaxonomic characteristics. Taxonomic data revealed the status of a separate genus for which the name Carnimonas gen. nov., sp., nov. is proposed. Despite chemotaxonomic and physiological similarities, the new genus is at present not a member of the family Halomonadaceae because of the lack of two out of 15 descriptive 165 rRNA signature sequences. The first member of the new genus is Carnimonas nigrificans. The use of a specific, 165 rRNA-targeted oligonucleotide primer allowed the identification of all nine strains of C. nigrificans in a PCR assay. Toxicological studies showed no pathogenic potential for C. nigrificans strain CTCBSIT (CECT 44379.
The results of DNA-DNA hybridization and chemotaxonomic studies indicated that the glutamic acid producers Brevibacterium divaricatum DSM 20297T (T = type strain), "Brevibacterium jlavum" DSM 20411, "Brevibacterium lactofermenturn" DSM 1412 and DSM 20412, Corynebacterium lilium DSM 20137T, and Corynebacterium glutamicum DSM 20300T and DSM 20163 are members of the same species. It is proposed that all of these strains should be classified in the species Corynebacterium glutamicum. Another glutamic acid-producing strain, Corynebacterium callunae DSM 20147T, was not related at the species level to C . glutamicum and should retain its separate species status. A restriction fragment length polymorphism analysis in which oligonucleotides targeted against conserved regions of 16s and 23s rRNA genes were used as hybridizing probes distinguished the individual strains. This method may be a helpful tool for strain identification.
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