Hypersensitivity pneumonitis due to metal working fluids: Detection of specific IgG antibodies to microbial antigens.

Occupational exposure to microbially contaminated metal working fluids (MWF) can cause hypersensitivity pneumonitis (HP). An important step in the diagnosis of HP is to identify the triggering antigen by detection of corresponding specific IgG antibodies (sIgG). As commercial sIgG tests are currently not available, protein antigens were prepared from MWF-workplace samples and from MWF-typical bacterial isolates. In 57 % of suspected HP-cases (n = 30) elevated sIgG concentrations were measured to at least one MWF-relevant antigen, of which Mycobacterium immunogenum was most prominent (88 %), followed by Pseudomonas oleovorans and Pseudomonas spec (82 % each), MWF-antigen mix and Pseudomonas alcaliphila (65 % each). Elevated sIgG concentrations to other microorganisms were measured to Micropolyspora faeni (82 %) and Aureobasidium pullulans (77 %). Correlation of sIgG values of all tested microbial antigens showed a significant relationship of MWF-antigen mixture to Pseudomonas antigens, but a low correlation to moulds. These newly prepared MWF-antigens are useful tools for the diagnosis of patients with suspected MWF-HP and are available for further investigations.

External validation of recombinant antigens for serodiagnosis of machine operator's lung.

BACKGROUND: Machine operator's lung (MOL) is a hypersensitivity pneumonitis the diagnosis of which is difficult. Our laboratory previously developed an ELISA test using recombinant antigens from Mycobacterium immunogenum isolated in French plant. The objective was to validate the previous ELISA results with ten new suspected cases from Germany. METHODS: Two serological analyses were performed: ELISA with the six recombinant antigens, and electrosyneresis with crude antigens of M. immunogenum and three other main species isolated from contaminated metalworking fluids. RESULTS: The two recombinant antigens acyl-CoA dehydrogenase and dihydrolipoyl dehydrogenase, combined together, and electrosyneresis are useful in making the diagnosis regardless of the clinical and radiological data. Finally 9 out of the 10 suspected cases were declared as MOL. CONCLUSIONS: 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.

Pseudochrobactrum lubricantis sp. nov., isolated from a metal-working fluid.

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)omega7c and C(19 : 0) cyclo omega8c), 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) (=CCUG 56963(T)=CCM 7581(T)).

Tessaracoccus lubricantis sp. nov., isolated from a metalworking fluid.

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) (=DSM 19926(T) =CCUG 55516(T)).

Corynebacterium lubricantis sp. nov., isolated from a coolant lubricant.

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 acids - tuberculostearic acid (10-methyl C(18 : 0)), C(16 : 0) and C(18 : 1)omega9c] 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) =DSM 45231(T) =CCUG 56567(T) =CCM 7546(T)).