Uric acid utilization by Mycobacterium intracellulare and Mycobacterium scrofulaceum isolates

Forty-nine human and environmental isolates of Mycobacterium intracellulare and Mycobacterium scrofulaceum were tested for their ability to grow on uric acid and a number of its degradation products. Nearly all (88 to 90%) strains used uric acid or allantoin as a sole nitrogen source; fewer (47 to 69%) used allantoate, urea, or possibly ureidoglycollate. Enzymatic activities of one representative isolate demonstrated the existence of a uric acid degradation pathway resembling that in other aerobic microorganisms.     

Genotyping Mycobacterium ulcerans and Mycobacterium marinum by using mycobacterial interspersed repetitive units

A novel category of variable tandem repeats (VNTR) called mycobacterial interspersed repetitive units (MIRUs) has been identified for Mycobacterium ulcerans (n = 39), M. marinum (n = 27), and one related organism. Fifteen MIRU loci were identified in the genome of M. marinum and were used to genotype M. ulcerans, M. marinum, and an M. marinum-like organism that is considered a possible missing link between M. marinum and M. ulcerans. Seven MIRU loci were polymorphic, and locus-specific PCRs for four of these loci differentiated seven M. ulcerans genotypes, four M. marinum genotypes, and a unique genotype for the missing link organism. The seven M. ulcerans genotypes were related to six different geographic origins of isolates. All isolates from West and Central Africa, including old and recent isolates, belonged to the same genotype, emphasizing the great spatiotemporal homogeneity among African isolates. Unlike the M. ulcerans genotypes, the four M. marinum genotypes could not be clearly related to the geographic origins of the isolates. According to MIRU-VNTR typing, all M. ulcerans and M. marinum isolates of American origin were closely related, suggesting a common American ancestor for these two pathogenic species on the American continents. MIRU typing has significant potential value for discriminating between reoccurrence and reinfection for M. ulcerans disease.     

Differentiation between Mycobacterium tuberculosis and Mycobacterium bovis by a multiplex-polymerase chain reaction

A multiplex-polymerase chain reaction (PCR) assay, based on one-step amplification and detection of three different mycobacterial genomic fragments, was designed for differentiation between Mycobacterium bovis and Mycobacterium tuberculosis. The oligonucleotide primers were chosen from the gro EL gene, present in the genus Mycobacterium sp., from the IS 6110 insertion sequence, present in Myco. tuberculosis complex and from the mtp 40 gene, identified as a specificspecies Myco. tuberculosis genomic fragment. This amplification method allowed the detection of two fragments of 576 and 317 base pairs in Myco. bovis and three fragments of 576, 396 and 317 base pairs in Myco. tuberculosis strains, including atypical strains of Myco. tuberculosis where the copy number of the IS 6110 element is low. The multiplex-PCR assay described may be a very useful tool for the rapid and specific differentiation of these related mycobacteria and easy to use in medical and veterinary microbiology laboratories.     

Cellular immunity in experimental infections caused by Mycobacterium avium and Mycobacterium scrofulaceum

The cell-mediated immunity was assessed in guinea pigs with experimental infection induced by Mycobacterium avium and M. scrofulaceum. It is established that the cell immunity indices in vitro correlate with the spread and gravity of the pathological process as well as with the tension of the immunological reactivity of the organism.     

Classification of Mycobacterium farcinogenes and Mycobacterium senegalense by immunodiffusion and thin-layer chromatography of long-chain components

Comparative immunodiffusion studies and thin-layer chromatographic analyses of whole-organism acid methanolysates were performed on 37 strains of Mycobacterium farcinogenes, Mycobacterium senegalense and Nocardia farcinica. The latter were clearly distinguished from the mycobacteria in containing a single mycolic acid methyl ester and showing more precipitinogens with nocardial than with mycobacterial and rhodococcal reference systems. The distribution of precipitinogens showed that M. farcinogenes and M. senegalense were very closely related and that both showed a greater affinity to Mycobacterium fortuitum than to any of the other established species of Mycobacterium tested. The complex pattern of alpha-mycolates and characteristic polar mycolates found in both M. farcinogenes and M. senegalense has only previously been found in M. fortuitum and Mycobacterium smegmatis.     

Partial characterization of Mycobacterium fortuitum and Mycobacterium smegmatis auxotrophs by syntrophism using Bacillus subtilis

Syntrophism (cross-feeding) could be demonstrated between mutants of Mycobacterium fortuitum and Mycobacterium smegmatis, and previously characterized mutants of Bacillus subtilis, auxotrophic for arginine, histidine, lysine or phenylalanine. Based on this cross-feeding data, the possible site of blockage in the biosynthetic pathways of the mutants could be inferred.     

Mycobacterium avium and Mycobacterium intracellulare chromatotypes defined by curvilinear gradient HPLC of mycolic acids

Seventy-nine strains of Mycobacterium avium complex bacteria (MAC), previously characterized by genetic probe analysis, were assayed using two methods of reverse phase high-performance liquid chromatography (HPLC) that employed curvilinear gradients. Although different in column length and cycle time, the methods produced equivalent results, yielding seven distinct chromatographic patterns (chromatotypes) of M. avium and M. intracellulare based on the ratio of mycolate concentrations in the late vs. the middle of three peak clusters (L:M ratio). The M. avium strains (n = 36) were assigned to chromatotypes 1 through 4 (L:M ratios less than 3), and the M. intracellulare strains (n = 25) to chromatotypes 5 through 7 (L:M ratios greater than 4). Of 18 Mycobacterium 'X' strains, seven resembled M. avium, seven others resembled M. intracellulare, and four were intermediate between M. avium and M. intracellulare.     

Rifampicin inactivation by Rhodococcus and Mycobacterium species

When prokaryotes are exposed to inhibitory concentrations of the antibiotic rifampicin, the only means hitherto identified by which cells overcome this inhibition is through mutational alteration in the target moiety, DNA-dependent RNA polymerase. In the nocardioform bacterium Rhodococcus erythropolis a novel mechanism has been identified, consisting of an inducible rifampicin-inactivating mechanism. Changes in the drug absorbance spectrum paralleled the decline in bacteriostatic activity of the antibiotic.     

Quantitation of seven elements in Mycobacterium phlei and Mycobacterium bovis by neutron activation analysis.

Quantitative determination of the elements potassium, sodium, manganese, magnesium, iron, cobalt and zinc was performed in mycobacteria by neutron activation analysis. Mycobacterium phlei ATCC 19 249 at different phase of growth (4, 8, 13, 23 and 37 days old cultures), and 14 days old Mycobacterium bovis BCG cultures and uninoculated semi-synthetic Sauton culture media were examined. The elements studied could be divided into three groups; sodium, potassium and magnesium could be regarded as major, iron as minor, and zinc, manganese and cobalt as trace elements. M. phlei contained, with the exception of zinc, higher amounts of elements than M. bovis. Other metals (aluminium, antimony, rubidium) could also be detected.     

Differentiation Between Mycobacterium kansasii and Mycobacterium marinum by Gas-Liquid Chromatographic Analysis of Cellular Fatty Acids

Comparison of the cellular fatty acids of 10 strains of Mycobacterium marinum and 35 strains of Mycobacterium kansasii revealed similarities within each species but differences between these two photochromogenic mycobacteria. A branched-chain fatty acid characteristic of M. kansasii was found in trace amounts in 2 of the 10 strains of M. marinum.     

Streptomycin Uptake by Mycobacterium tuberculosis

Data are presented indicating that (14)C-streptomycin uptake by tubercle bacilli increases as a function of exposure time, is directly and proportionately related to concentration, and involves at least two phases.     

Cholesterol metabolism by Mycobacterium.

Cholesterol metabolism by Mycobacterium species ATCC Number 19652 was studied in defined media. Whole cells were found to take up 91% of the total cholesterol when incubated five days at 34 degrees C in media of pH 6.8-7.4. Uptake of cholesterol by whole cells could be significantly inhibited by 2,4-dinitrophenol and dicyclohexylcarbodiimide. Growth media supernates as well as isolated microbial cell walls were found to contain cholesterol hydrolysing activity. This activity was extractable by Triton X-100 and appeared to have a molecular weight of approximately 100-200,000.     

Expression profiles of low molecular mass heat shock proteins by Mycobacterium avium and Mycobacterium intracellulare

Closely related non-tuberculous mycobacterial species, Mycobacterium avium and Mycobacterium intracellulare, were compared for the profiles of their production of low molecular mass heat shock proteins at 45 degrees C, by performing polyacrylamide gel electrophoresis analysis of bacterial cell lysate proteins. All of the M. intracellulare but not M. avium strains potently increased the production of the 18-kDa heat shock protein, when cultured at 45 degrees C. Half of the M. intracellulare strains with high sensitivity to 45 degrees C produced not only the 18-kDa heat shock protein but also the 16-kDa heat shock protein at 45 degrees C. These findings indicate that M. avium and M. intracellulare differentially respond to 45 degrees C heat shock in terms of the production of low molecular mass heat shock proteins.