A Gram-positive-staining, coccoid-shaped, oxidase-negative, non-spore-forming, non-motile bacterium, strain KSS-17Se T , was isolated from a metalworking fluid. On the basis of its major fatty acid (ai-C 15 : 0 ) and 16S rRNA gene sequence similarity, the strain grouped with Tessaracoccus bendigoensis and Tessaracoccus flavescens, sharing 95.3 and 97.4 % 16S rRNA gene sequence similarity with the respective type strains. Similarities with other established species of the genera Luteococcus, Propioniferax and Granulicoccus were lower than 95.5 %. The quinone system was characterized by the major menaquinone MK-9(H 4 ). In the polar lipid profile, diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid and an unknown polar lipid were detected as major compounds. Additionally, three unknown glycolipids and minor amounts of phosphatidylethanolamine, two unknown aminolipids and two unknown polar lipids were detected. Phosphatidylinositol was present only in trace amounts. Predominant polyamines were spermine and spermidine. LL-Diaminopimelic acid was identified as the diagnostic diamino acid in the cell wall. The strain showed clear differences in phenotype (including chemotaxonomic features) from both Tessaracoccus species and members of the other above-mentioned genera. DNA-DNA hybridization between KSS-17Se T and T. bendigoensis Ben-106 T and T. flavescens SST-39 T yielded similarities of 15.1 and 21.0 %, respectively. It is evident that the organism represents a novel species, for which the name Tessaracoccus lubricantis sp. nov. is proposed. The type strain is KSS-17Se T (5DSM 19926 T 5CCUG 55516 T ).Metalworking fluids are substances that are used for cooling and lubrication when metals are being cut or formed. A distinction is drawn between non-watermiscible and water-miscible metalworking fluids; the latter are products which are mixed with water before use. In 2007, more than 32 000 tons of water-miscible metalworking fluid concentrate were used in Germany. Assuming a 4 % batch, this means an estimated consumption of 800 000 tons of emulsion or solution per year.The investigations described here were all done with oleaginous, preserved, water-mixed metalworking fluids. In a study on the diversity of bacteria isolated from different metalworking fluids, strain KSS-17Se

Three Gram-positive, rod-shaped, oxidase-negative, non-spore-forming, non-motile bacteria (KSS-3Se T , KSS-4Se and KSS-10Se) were isolated from a coolant lubricant. 16S rRNA gene sequence analyses revealed almost identical sequences, with only a few (,10 positions) differences for these three isolates. Comparisons showed the highest similarities to Corynebacterium pilosum NCTC 11862 T (97.6 % similarity with strain KSS-3Se T ). Similarities with other established Corynebacterium species were lower than 97.0 %. Chemotaxonomic data studied for strain KSS-3Se T [polar lipids -major compounds phosphatidylglycerol and an unknown glycolipid, moderate amounts of phosphatidylinositol and diphosphatidylglycerol; polyamines (small amounts) -major compounds spermidine and spermine; quinones -significant amounts of menaquinones MK-9(H 2 ), MK-8(H 2 ) and MK-7(H 2 ); and major fatty acidstuberculostearic acid (10-methyl C 18 : 0 ), C 16 : 0 and C 18 : 1 v9c] were congruent with those reported for Corynebacterium. The strain showed differences in phenotype from C. pilosum. DNA-DNA hybridizations between KSS-3Se T and C. pilosum DSM 20521 T yielded a relatedness of 22.9 % (20.4 % in the reciprocal assay). From these results, it is evident that the organisms represent a novel species, for which the name Corynebacterium lubricantis sp. nov. is proposed (type strain KSS-3Se T 5DSM 45231 T 5CCUG 56567 T 5CCM 7546 T ).

A Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile bacterium (KSS 7.8 T ) was isolated from a water-mixed metal-working fluid. On the basis of 16S rRNA gene and recA sequence similarities, the isolate was clearly grouped in the genus Pseudochrobactrum. This allocation was confirmed by fatty acid data (major fatty acids: C 18 : 2 v7c and C 19 : 0 cyclo v8c), polar lipid profile (major components: phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine, plus moderate amounts of phosphatidylmonomethylethanolamine and unknown aminolipid AL1), quinone system (ubiquinone Q-10) and polyamine pattern (spermidine and putrescine predominant). DNA-DNA pairing with the most closely related Pseudochrobactrum species showed values ranging from 24.2 to 45.7 %, and physiological and biochemical data clearly differentiated this isolate from described Pseudochrobactrum species. This organism represents a novel species of the genus Pseudochrobactrum, for which the name Pseudochrobactrum lubricantis sp. nov. is proposed, with the type strain KSS 7.8 T (5CCUG 56963 T 5CCM 7581 T ).The genus Pseudochrobactrum was proposed by Kämpfer et al. (2006). At present, it comprises Pseudochrobactrum saccharolyticum, Pseudochrobactrum asaccharolyticum, Pseudochrobactrum kiredjianiae (Kämpfer et al., 2006(Kämpfer et al., , 2007 and Pseudochrobactum glaciei (Romanenko et al., 2008). Members of the genus can be clearly differentiated from members of the genera Ochrobactrum and Brucella on the basis of 16S rRNA gene sequence and recA sequence data, as well as chemotaxonomic data (Kämpfer et al., 2006(Kämpfer et al., , 2007.Strain KSS 7.8 T was isolated from a metal-working fluid of a metal-processing company in Germany on nutrient agar at 30 u C. Subcultivation was done on tryptone soy agar at 28 uC for 48 h. On this agar, this organism grew also at 15-45 u C, but not at 10 or 50 u C. Growth at 30 u C was also observed on MacConkey agar and R2A agar (all from Oxoid).Gram staining was performed as described by Gerhardt et al. (1994). Cell morphology was observed under a Zeiss light microscope at 61000, with cells grown for 3 days at 30 u C on nutrient agar. The 16S rRNA gene was analysed as described previously (Kämpfer et al., 2003). Sequence analysis and similarity calculations were carried out by using the software programs Bionumerics v. 4.0 (Applied Maths BVBA) and MEGA v. 3.1 (Kumar et al., 2004). The sequenced length of the 16S rRNA gene was 1401 bp (GenBank accession no. FM209496) for strain KSS 7.8 T . Sequence similarities to the three most closely related Pseudochrobactrum species were 99.8 % to P. saccharolyticum CCUG 33852 T , 99.8 % to P. asaccharolyticum CCUG 46016 T and 99.3 % to P. kiredjianiae CCUG 49584 T . A lower similarity of 96.2 % was found to P. glaciei KMM 3858 T . The 16S rRNA gene-based tree shown in Fig. 1 results from a neighbour-joining reconstruction using the Kimura two-parameter correction and 1000 resamplings for bootstrap analysis.The partial r...

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Despite the geographical distance, the crude and recombinant antigens produced to investigate the clustered French cases also proved to be useful in diagnosing the suspected cases in Germany.