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The virulence for mice of some newly described slowly growing mycobacterial species, M. intermedium DSM 44049 (Group I), M. lentiflavum ATCC 51985 and M. interjectum ATCC 51457 (Group II), M. genavense ATCC 51233, M. celatum ATCC 51131, M. branderi ATCC 51788 and M. conspicuum DSM 6333 (Group III) was compared with the virulence of M. intracellulare N-260 (virulent strain). 10(6) CFU cultured in Middlebrook 7H9 medium were given i.v. to the BALB/c mice. Infected animals were sacrificed after one day and 56 days, and observed for the incidence and/or degree of gross lesions of visceral organs. The bacterial load (CFU) in tissue homogenates of the lungs and spleen was counted using 7H11 agar plates. Macroscopically, marked enlargement of the spleen and numerous small whitish nodules in the lungs of mice infected with M. intracellulare were noted, whereas with other test strains no gross disease in the lungs and slight enlargement of the spleen of mice infected with M. genavense and M. branderi were noted. The average log CFU in the lungs and spleen 56 days after the infection was in the order as follows: M. intracellulare (5.90 and 7.57, respectively), M. genavense (4.35, 7.15), M. celatum (5.01, 6.00), M. branderi (4.09, 6.33), M. conspicuum (3.11, 5.10), M. intermedium (2.26, 4.19), M. lentiflavum (1.70, 3.91), and M. interjectum (1.43, 2.93). Microscopically, granulomatous lesions mainly consisted of macrophages with acid-fast bacilli were found in the alveolar walls and splenic red pulp of mice infected with M. intracellulare. No histopathological changes were produced in the lungs of any of mice infected with other test strains, and some milder features were revealed in the spleen of mice infected with M. genavense, M. branderi and M. celatum. Thus, we concluded that all of the test strains of the newly described nontuberculous slowly growing mycobacteria were less virulent for mice than were M. intracellulare.

The "24-HOUR BATH" is an apparatus which circulates the bath water, keeps it clean and warm, and makes it possible to take a bath at any time during the day or night. It consists of apparatus for cleaning (sponge or mesh filter and filter material), heating (ceramic heater), and sterilizing (UV lamp). Recently, three cases of skin disease due to M. avium infection in private homes, in which "24-HOUR BATH" water was suspected to be the source of infection, have been reported. We attempted to isolate M. avium complex from the water (32 specimens), sponge filter (29 specimens), and filter material (32 specimens) of the "24-HOUR BATH". One hundred-ml samples of bath water, and 50-ml samples of rinse from a sponge filter or filter material were centrifuged at 3000 rpm for 20 min. Sediment was suspended in distilled water and a smear was prepared, and then digested and decontaminated with 2% sodium hydroxide. The processed specimens were cultured on 2% Ogawa medium containing ofloxacin (1 microgram/ml) and ethambutol (2.5 micrograms/ml) for 8 weeks at 37 degrees C. Positive smears were 3 (9.4%), 25 (86.2%) and 25 (78.1%) specimens from the water, sponge and filter material, respectively. A few bacterial clumps were observed, especially in the sponge specimens. The number of positive culture was 5 (15.6%), 24 (82.8%) and 25 (78.1%) from the water, sponge and filter material, respectively. Among them the number of Runyon's Group III-positive cultures was 5 (100%), 22 (91.7%) and 20 (80%) in the water, sponge, and filter material specimens, respectively. In most cases, cultures were positive for both the sponge and filter material specimens. All of the Group III mycobacteria were smooth, grew at 28, 37, 42, and 45 degrees C, negative for niacin, nitrate reductase, semiquantitative catalase, urease and Tween80 hydrolysis, and positive for 68 degrees C catalase. All of the strains reacted with M. avium complex AccuProbe and M. avium AccuProbe, but none of the strains reacted with M. intracellulare AccuProbe. Therefore, all the Group III isolates were identified as M. avium by the culture, biochemical and genetical characteristics.

M. avium complex (MAC) is one of the important causative agents of opportunistic infections among AIDS patients. Recent evidence showed that the entry of infection is through the gastrointestinal tract. In the present study, we compared the prophylactic effect of some antimicrobials against MAC infection induced in mice. Different groups of beige mice were fed with pellets containing 0.0067% (10 mg/kg) of KRM-1648, rifabutin (RFB) and clarithromycin (CAM). Seven days after the administration of drugs, the mice were infected with M. intracellular N-241 (5 x 10(8) CFU) orally, five times, every other day and killed one and 126 days after the last infection. The effect of drug was evaluated using the frequency and severity of gross lung lesions in the mice and by the total CFU recovered from the lungs and spleen. MAC infection was not likely to be established since there was no macroscopic evidence of lesion in organs and the recovery of cultures from lungs and spleen tested was negative, in 3 of 10 mice in the control group, 2 of 9 in the CAM group, 4 of 9 in the RFB group and 4 of 10 in the KRM group. These mice were excluded from the analysis of the study results. Thus, we examined 7 mice in the control group, 7 in the CAM group, 5 in the RFB group, and 6 in the KRM group. Tubercle-like lesions were observed in the lungs of all 7 mice in the control group (severity: 3+ in 5 mice; 4+ in 2 mice), in 5 of 7 mice (71%) in the CAM group (severity: 2+ in 1 mouse; 3+ in 4 mice), and in 4 of 5 mice (80%) in the RFB group (severity: 1+ in 1 mouse; 2+ in 1 mouse; 4+ in 2 mice), while only slight lesions (severity: 1+) were observed in 4 of 6 mice (67%) in the KRM group. There was no macroscopic evidence of lesion in spleen, liver and kidneys. The log CFU was determined at the next day of the completion of the last infection. The log CFU of the lungs was 2.49 and 2.28 in the control group and the CAM group, respectively. The bacteria were not recovered either from the lungs in the RFB and KRM groups, nor from the spleen in all the groups. The order of efficacy of the drugs on the basis of the CFUs recovered from the lungs and spleen in each group determined 126 days after the completion of the last infection was as follows; KRM > CAM > RFB in the lungs and KRM > CAM [symbol: see text] RFB in the spleen, although there was no significant difference among the three drugs (P < 0.05). However, the significantly preferable effect was obtained in the three drug groups as compared with the control group.

The BBL MGIT Mycobacteria Growth Indicator Tube is a novel broth based culture system for the detection of mycobacteria from clinical specimens. The tubes consist of a fluorescent indicator embedded in silicone on the bottom of a 16x100 mm round-bottom tube, filled with 4ml of an enriched BBL Middlebrook 7H9 broth base, with 0.25% glycerol. Actively growing mycobacteria consume the oxygen dissolved in the medium and fluorescence will occur when the tubes are observed with a 365nm transilluminator. The purpose of this study is to evaluate comparatively MGIT with 1% Ogawa egg medium by using two hundred and forty-five clinical specimens. The samples were digested, decontaminated and concentrated for culture using N-acetyl-L-cysteine-sodium hydroxide method. Fifty-nine of 245 (24%) clinical samples were culture positive for mycobacteria (43 M. tuberculosis complex, 12 M. avium complex and 4 other species) by one or both test systems. The MGIT detected 4 isolates of M. tuberculosis complex and 6 isolates of M. avium complex not recovered by the Ogawa egg medium, respectively. The mean time of detection of M. tuberculosis complex was 13 days (range: 2-26 days) and 19 days (range: 8-31 days) for MGIT and Ogawa egg medium, respectively, and that of M. avium complex was 5 days (range: 2-8 days) and 16 days (range: 6-22 days) for the MGIT and Ogawa egg medium, respectively. Overall, the MGIT is a sensitive culture system for the detection of mycobacteria from clinical specimens, is easy to use and may be applicable to clinical laboratories.