Ten new Legionella species were characterized on the basis of biochemical reactions, antigens, cellular fatty acids, isoprenoid quinones, and deoxyribonucleic acid relatedness. Nine of the new species were isolated from the environment, and one, Legionella hackeliae, was isolated from a bronchial biopsy specimen obtained from a patient with pneumonia. The species all exhibited the following biochemical reactions typical of the legionellae: growth on buffered cysteine-yeast extract agar, but not on blood agar; growth requirement for cysteine; gram negative; nitrate negative; urease negative; nonfermentative; catalase positive; production of a brown pigment on tyrosine-containing yeast extract agar; liquefaction of gelatin; and motility. Legionella s4iritensis was weakly positive for hydrolysis of hippurate; the other species were hippurate negative. Legionella cherrii, Legionella steigerwaltii, and Legionella parisiensis exhibited bluish white autofluorescence. Legionella rubrilucens and Legionella erythra exhibited red aqtofluorescence. The other species, L. spiritensis, L . hacke liae, Legionella maceachernii, Legionella jamestowniensis, and Legionella santicrucis did not auto fluoresce bluish white or red. All species had cellular fatty acid contents qualitatively similar to those of previously described legionellae and had major amounts of ubiquinones with more than 10 isoprene units in the side chains. Each new species was serologically distinct from previously described Legionella species. As determined by the hydroxyapatite method at 60°C, two strains of L. maceachernii were 100% related, and four strains of L. cherrii were 94 to 99% related. The other new species were represented by single strains. The levels of relatedness of the new species to each other and to previously described legionellae ranged from 1 to 67%. L . maceachernii, L. japestowniensis, and L. hackeliae were less than 25% related to other species. L. rubrilucens and L. erythra, and two red-autofluorescing species, were about 60% interrelated. L. spiritensis (a non-autofluorescing species) was 34% related to L. rubrilucens. L . santicrucis was 64% related to Legionella sainthelensi. The three bluish white-autofluorescing species, L. parisiensis, L. cherrii, and L. steigerwaltii, were most closely related to other bluish white-autofluorescing species, especially Legionella bozemanii, Legionella dumofli, Legionella gormanii, and "Legionella anisa" (35 to 67%).

No abstract

Sets of sera (444) submitted for diagnostic testing for legionellosis were tested against 29 indirect immunofluorescence assay (IFA) antigens prepared from the characterized Legionella species and Legionella-like organisms to determine the prevalence of antibodies to Legionella organisms. Reciprocal titers of 15% of the serum sets rose fourfold or more to greater than or equal to 128 (indicating seroconversion) against one or more Legionella antigens. The specificity of the test was 96% when evaluated in patients with pneumonia due to non-Legionella organisms. Antibodies were of the IgG, IgM, and (infrequently) IgA classes and were either specific for a single species (as defined by a difference in titer of fourfold or more) or reacted with common Legionella antigens (30 [45%] vs. 36 [55%] of 66 seroconversions, respectively). No single antigen detected half of the positive sera. Elevated IFA titers (of greater than or equal to 256) against single or multiple Legionella antigens occurred in 12% of 184 normal control sera. Therefore, only seroconversions to titers of greater than or equal to 128 should be considered indicative of recent Legionella infection.

A semiautomated, kinetic-dependent, enzyme-linked immunosorbent assay (K-ELISA) was adapted to detect serum antibodies to Legionella pnellmophila. In a comparative study, 158 human serum samples (79 pairs) were tested by K-ELISA and the standard indirect immunofluorescence assay for determination of antibody levels to L. pneiumophila serogroup 1. K-ELISA determinations were made by using a serogroup-specific antigen or a preparation (unfractionated antigen) which contained both common antigen and serogroup-specific reactivity. There was good correlation between the immunofluorescence assay and the K-ELISA by using either antigen, although greater correlation was achieved with the unfractionated antigen (coefficients of correlation, 0.894 with unfractionated antigen and 0.841 with serogroup-specific antigen). These results indicate that the K-ELISA is a reliable alternative to the immunofluorescence assay for serologically diagnosing legionellosis.

Indirect immunofluorescence assay titers of human sera obtained against the Legionella pneumophila serogroup 1 antigen (Philadelphia 1 strain) killed with 10% Formalin showed a tendency to be lower than those obtained against the reference heat-killed antigen (geometric mean titer of 194 and 370, respectively) when all other variables in the test were held constant. Test results were interpreted the same for 96% of 60 paired sera if the cutoff level used to interpret a positive test result for the formalinized antigen was lowered by one twofold dilution factor.