The aerobic actinomycetes are soil-inhabiting microorganisms that occur worldwide. In 1888, Nocard first recognized the pathogenic potential of this group of microorganisms. Since then, several aerobic actinomycetes have been a major source of interest for the commercial drug industry and have proved to be extremely useful microorganisms for producing novel antimicrobial agents. They have also been well known as potential veterinary pathogens affecting many different animal species. The medically important aerobic actinomycetes may cause significant morbidity and mortality, in particular in highly susceptible severely immunocompromised patients, including transplant recipients and patients infected with human immunodeficiency virus. However, the diagnosis of these infections may be difficult, and effective antimicrobial therapy may be complicated by antimicrobial resistance. The taxonomy of these microorganisms has been problematic. In recent revisions of their classification, new pathogenic species have been recognized. The development of additional and more reliable diagnostic tests and of a standardized method for antimicrobial susceptibility testing and the application of molecular techniques for the diagnosis and subtyping of these microorganisms are needed to better diagnose and treat infected patients and to identify effective control measures for these unusual pathogens. We review the epidemiology and microbiology of the major medically important aerobic actinomycetes.

A recent study of Nocardia asteroides revealed that 95% of clinical strains had one of five antibiotic resistance patterns. We found the pattern of resistance to cefotaxime and cefamandole in 19% of 200 clinical N. asteroides isolates. Isolates with this drug resistance pattern were from numerous geographic sources and were associated with significant clinical disease (56% of patients had disseminated infections). Phenotypic studies revealed that these isolates were relatively homogeneous and matched previous descriptions and reference strains of the controversial species N. farcinica. Growth at 45 degrees C, acid production from rhamnose, ability to utilize acetamide as a nitrogen and carbon source, and resistance to tobramycin and cefamandole were features of N. farcinica that could be tested in the clinical laboratory and allowed their distinction from N. asteroides. The serious nature of disease due to N. farcinica and its resistance to the newer cephalosporins suggest a clinical need for laboratory identification of this species.(Current tests used in clinical laboratories do not distinguish N. farcinica from N. asteroides.) This is the first recognition that N. farcinica has a specific drug resistance pattern and confirms the previously described concept that drug resistance patterns of N. asteroides may be associated with specific taxonomic groups.

Agar media made with 0.4% colloidal chitin plus mineral salts and adjusted to pH 8.0 was superior to four other commonly used media for the isolation and enumeration of actinomycetes from water samples. More actinomycetes developed on chitin agar, and the development of bacteria and fungi was suppressed. Frozen and vacuum-dried chitin from aqueous colloidal suspensions was finely divided and gave results comparable to those obtained with media prepared from colloidal suspensions.

A commercial DNA probe procedure for identification of Mycobacterium avium complex isolates was evaluated for accuracy and applicability for use in the clinical laboratory. The test (Gen-Probe Rapid Diagnostic System for Mycobacterium Avium Complex; Gen-Probe Corp., San Diego, Calif.) uses hybridization in solution of two 125I-labeled cDNA probes. One probe is complementary to rRNA from M. avium, and the other is complementary to rRNA from M. intracellulare. Results are expressed as absolute percent hybridization, with values greater than or equal to 10% considered positive. The procedure accurately identified all 134 M. avium complex isolates and concomitantly identified them to species level. There were no false-positives with 66 other mycobacteria, including 22 M. tuberculosis and 18 M. kansasii isolates, or with 8 Nocardia isolates. The mean percent hybridization (+/- the standard deviation) of M. avium probe-positive isolates (94 isolates) was 48.0 +/- 9.9 (range, 11.5 to 72.7); for M. intracellulare (40 isolates), it was 45.7 +/- 8.8 (range, 22.7 to 60.7). Among the 74 non-M. avium complex isolates, the percent hybridization range was 1.0 to 4.2, except for a single value of 9.7 which was less than 5 when the test was repeated. Four M. avium complex isolates reacted positively with both probes on initial testing, and three were confirmed. On repeat testing of subcultures, each reacted with only one probe, suggesting the presence of a mixed culture. The procedure can be completed in as little as 2 h and could easily be performed in most clinical laboratories.

During a taxonomic investigation of Nocardia spp. and Streptomyces spp., we received 658 isolates from laboratories of both human and veterinary medicine. Our procedure leading to the identification of 92% of these isolates, the species that they represented, and a characteristic pattern of properties of the strains of these species are presented. A key devised for the tentative identification of species of nocardiae and streptomycetes that are believed to be of medical importance is included to assist clinical microbiologists in identifying their strains.

Nineteen reference and 156 clinical strains of the genus Nocardia belonging to 12 taxonomic groups were studied for restriction fragment length polymorphism (RFLP) by using an amplified 439-bp segment of the 65-kDa heat shock protein gene. Of 30 restriction endonucleases, digestion with MspI and then digestion with BsaHI produced RFLP band patterns which separated all 12 groups except N. asteroides type IV from 6 of 12 N. transvalensis isolates and N. carnea from the N. asteroides type VI isolates. Commonly encountered species such as N. nova, N. farcinica, N. brasiliensis sensu stricto, and N. otitidiscaviarum were easily separated. Each taxon resulted in a single RFLP band pattern that included > or = 96% of all biochemically grouped isolates for 9 of 12 taxa with MspI and for 8 of 12 taxa with BsaHI. With the use of both patterns, only 6 of 175 (3.4%) isolates failed to fit the biochemically defined group patterns. These studies provide the first evidence for the separate identities of four antibiogram-defined (but currently unnamed) groups within the N. asteroides complex (types I, II, IV, and VI) and the presence of two subgroups within N. transvalensis. They also provide genotypic evidence for the separate identities of N. nova and N. farcinica. The lack of BstEII recognition sites in amplicons obtained from nocardiae provides a simple and rapid method for the differentiation of nocardiae from mycobacteria. DNA amplification with RFLP analysis is the first rapid method that distinguishes all clinically significant taxa and recognized species within the genus Nocardia.