New methods for the isolation of Legionella pneumophila.

Abstract

Some new methods for the isolation of Legionella pneumophila are described which have been successful in recovering this organism from 6/10 patients with clinical evidence of Legionnaires' disease. The increased sensitivity of these methods combined with speedier isolation of the organisms than has hitherto been possible will hopefully lead to eventual isolation of this organism as a routine procedure in diagnostic microbiology laboratories.

Pathological and Biochemical Features of Legionella Pneumophila Infection in Guinea-pigs

“…Because the strain of L. pneumophila in tissue homogenate did not grow on charcoal-yeast extract agar (Feeley et al, 1979) or enriched blood-agar (Greaves, 1980), the presence of the organism was confirmed by indirect fluorescent-antibody staining with antiserum to the organism produced in rabbits and a goat anti-rabbit serum labelled with fluorescein isothiocyanate.…”

Identification of Legionella Pneumophila Antigens and Antibodies by Immunoferritin Electronmicroscopy

“…Philadelphia-2 organisms, grown on enriched blood agar (Greaves, 1980) for 3 days in 5% C02 at 37"C, were harvested and a thick suspension was prepared in PBS-A. To this was added formalin and a suspension of normal yolk-sac membrane in PBS-A to give final concentrations of 2% (v/v) and 0.5% (w/v) respectively.…”

“…Twenty-one strains of L. pneumophila comprising members ofthe six established serogroups were examined by direct EM and IFEM after cultivation on enriched blood agar (Greaves, 1980). These were: serogroup 1, strains Philadelphia 1 and 2, Pontiac 1, Cambridge 1, Washington 1, Nottingham N/P/l, N/M/2, N3, N/P/4, N5, N6, N7, N8, N9, Bloomington 1 and Corby 1; serogroup 2, strain Togus 1; serogroup 3, strain Bloomington 2; serogroup 4, strain Los Angeles 1; serogroup 5, strain Cambridge 2; and serogroup 6, strain Oxford 1.…”

“…The organisms are typically small gram-negative bacilli which have been isolated from post-mortem lung tissue on blood agar containing added iron and cysteine (Greaves, 1980), by intraperitoneal inoculation of guinea-pigs and yolk-sac inoculation of fertile hen's eggs.…”

Enhanced Antifungal Effect of the Selective Medium for the Detection of Legionella Species by a Combination of Cycloheximide, Amphotericin B and Thiabendazole

“…The addition of antifungal antibiotics such as (Greaves, 1980;Okuda et al, 1984), anisomycin (Edelstein, 1981;1982), cycloheximide (Bopp et al, 1981; and natamycin (Edelstein et al, 1996) Effect of antifungal agents against molds Next, we tested the effect of the antifungal agents in five types of selective agar plates on the growth of molds (Table 2). Both GVP a (without antifungal agents) and GVPG a (with griseofulvin) plates were overgrown with molds.…”