Charcoal-yeast extract agar is a new bacteriological medium that supports excellent growth of the Legionella pneumophila. It results from modifications made in an existing L. pneumophila medium, F-G agar. Yeast extract, instead of an acid hydrolysate of casein, serves as the protein source. Beef extractives and starch are not added. Activated charcoal (Norit A or Norit SG) is included at 0.20% (wt/vol). Comparison of charcoal-yeast extract and F-G agars showed that a greater number of colony-forming units of L. pneumophila was recovered from a standardized tissue inoculum on charcoal-yeast extract agar (4.35 x 106 colonyforning units) than on F-G agar (4.85 x 104 colony-forming units). Macroscopic colonies of L. pneumophila were visible on the new medium within 3 days, whereas 4 days of growth was required on F-G agar. McDade et al. initially isolated the Legionnaires disease bacterium, now named Legionella pneumophila, by using guinea pigs and embryonated chicken eggs (2, 9). For the purpose of growing L. pneumophila on artificial media, Weaver inoculated 17 different bacteriological agars with an L. pneumophila-infected yolk sac suspension. Mueller-Hinton agar supplemented with 1% hemoglobin and 1% IsoVitaleX (BBL
We analyzed 24 environmental samples collected in or near the Indiana Memorial Union, where an epidemic of Legionnaires' disease occurred in early 1978. We conducted fluorescent antibody analyses and culture on F-G and charcoal yeast extract agars of each sample directly; splenic tissue of guinea pigs inoculated with the sample; and yolk sacs from embryonated eggs inoculated with splenic tissue of guinea pigs injected with the sample. Legionnaires' disease (LD) bacterium was isolated from seven of the 24 samples: one water sample from the air-conditioner cooling tower of the Union; three water samples from a stream near the Union; and three mud samples from the same stream. The LD bacterium strains were of three different serotypes. These findings indicate that LD bacteria may be widespread in nature.
Epidemiological and microbiological studies were conducted in a hospital room with carpet (CR) and in one with carpet (NCR). Microbiological profiles were determined with specimens obtained from patients admitted to these rooms. Patient records were reviewed to note infection status and other case identities. Eleven-millimeter cylindrical core samples of carpet were obtained, and swab template techniques were used on the bare floor for subsequent enumeration and identification of contaminating microorganisms. In each sampling period, higher microbial counts per square inch (1 in(2) = ca 6.452 cm(2)) were measured for the carpet than for the bare floor. Recovery rates of Enterobacter spp., Klebsiella pneumoniae, and Escherichia coli were higher from carpet samples than from bare floor samples. Typable organisms (such as E. coli, Pseudomonas aeruginosa, K. pneumoniae, and Staphylococcus aureus) obtained from patients were also more frequently recovered from the carpet than from the bare flooring. Patients who stayed in the CR were shown to be colonized with the same types of organisms as those initially recovered from the carpet. However, no statistically significant differences were found in patients in the CR versus NCR in colonization with all typable and nontypable organisms first found on the floor. Disease in patients was found not to be associated with organisms found as contaminants of the carpet or the bare floor. Air above carpeting contained more consistent concentrations of organisms than air above the bare flooring.
Legionella pneumophila suspended in tap water was exposed to biocides recommended for inhibiting biological growth in cooling towers and evaporative condensers of air-conditioning systems. Chlorine, 2,2-dibromo-3-nitrilopropionamide, and a compound containing didecyldimethylammonium chloride and isopropanol were effective in destroying concentratiois of 10(5) to 10(6) viable cells per ml. Formulations consisting of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one, disodium ethylene bis(thiocarbamate) and sodium dimethyl dithiocarbamate, and a phenolic with pentachlorophenate and sodium salts of other chlorophenols were less effective.
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