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Determination of antimicrobial susceptibility patterns of Nocardia spp. from clinical specimens by Etest.

Susceptibilities to 11 antimicrobial agents were determined by Etest for 93 Nocardia isolates from clinical specimens and 15 type strains belonging to different Nocardia spp. All isolates were susceptible to trimethoprim-sulphamethoxazole, amikacin and linezolid, but susceptibilities of the various Nocardia spp. to beta-lactams, aminoglycosides, ciprofloxacin and clarithromycin varied markedly. Overall, there was a good correlation between the drug resistance patterns and the species identification established by conventional phenotypic tests and 16S rDNA sequencing. Among the different species encountered, Nocardia farcinica and Nocardia brasiliensis displayed the most multiresistant profiles, with resistance to imipenem occurring mainly among isolates of N. brasiliensis and Nocardia abscessus. The species variability in susceptibility profiles and the numerous recent taxonomic changes means that in-vitro susceptibility tests may be a complementary tool for the identification of Nocardia isolates from human clinical specimens. Further studies on a larger number of species from more diverse geographical sources, including species that are found less commonly among clinical isolates, are required to validate and extend the results.

Anti-Bacterial Agents↗

Isolation of Nocardia sp. from blood cultures in a teaching hospital.

To determine the clinical significance of the isolation of Nocardia sp. from blood cultures, clinical records of patients with bacteremia due to Nocardia sp. in our hospital from January 1986 to December 1990 were retrospectively reviewed. Eight patients had 9 blood cultures positive for Nocardia sp. Four of the isolates were identified as Nocardia asteroides. Only 2 of the patients had disseminated nocardiosis due to N. asteroides, and both were appropriately treated for Nocardia sp. In the other cases, isolation of Nocardia sp. was considered nonsignificant or undetermined, and all were clinically cured. Nocardia sp. may be an occasional contaminant of blood cultures. The significance of nocardemia is a matter of clinical judgment.

Adult↗

Heavy metal removal by activated sludge: influence of Nocardia amarae.

The goal of this research was to examine the metal binding capacity of Nocardia amarae cells and to assess the influence of Nocardia cells on the overall metal binding capacity of activated sludge. Metal sorption capacities of the pure Nocardia cells and activated sludge biomass containing various levels of added Nocardia pure cultures were determined by a series of batch experiments. Batch sorption isotherms for nickel (Ni), copper (Cu), and cadmium (Cd) showed that the pure culture of N. amarae exhibited significantly higher metal sorption capacity than the activated sludge biomass obtained from Wilmington Wastewater Treatment Plant (Wilmington, DE). Surface area of biomass estimated by a dye technique showed that pure N. amarae cells growing at stationary phase have substantially more specific surface area than that of activated sludge from Wilmington Treatment Plant. A two-fold difference in specific surface area indicated that the higher metal sorption capacity of Nocardia cells may be due to the higher specific surface area. The metal sorption capacity of activated sludge increased proportionally with the amount of Nocardia cells present in the mixed liquor. This increase was attributed to the greater specific surface area of the mixed liquor samples containing greater amounts of Nocardia cells.

Adsorption↗

Nocardia uniformis nom. rev.

A soil isolate representing the putatively novel species 'Nocardia uniformis' was found to have morphological, staining and chemotaxonomic properties consistent with its classification in the genus Nocardia. An almost complete sequence of the 16S rDNA of the strain was determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for nocardiae and phylogenetic trees were inferred using four tree-making algorithms. The organism was consistently associated with the type strain of Nocardia otitidiscaviarum albeit with a relatively low bootstrap value recorded for neighbour-joining analysis. The strain was also readily separated from representatives of all validly described Nocardia species using a set of phenotypic properties. The genotypic and phenotypic data indicate that the strain should be assigned to the genus Nocardia as a new species. The name proposed for the new species is Nocardia uniformis. The type strain is JCM 3224T.

Algorithms↗

Nocardia beijingensis sp. nov., a novel isolate from soil.

The taxonomic status of a soil isolate, provisionally assigned to the genus Nocardia, was established following a polyphasic study. An almost complete sequence of the 16S rDNA of the strain was determined and aligned with available sequences for nocardiae and phylogenetic trees were inferred using four tree-making algorithms. The organism was associated consistently with the type strains of Nocardia brasiliensis and Nocardia farcinica in the neighbour-joining analysis, albeit with low bootstrap values. It was also found to have chemotaxonomic and morphological properties consistent with its assignment to the genus Nocardia. The organism was readily separated from representatives of other Nocardia species by a broad range of phenotypic properties. The name proposed for the novel species is Nocardia beijingensis sp. nov. The type strain is AS4.1521T (= IFO 16342T = JCM 10666T).

Base Composition↗

Nocardia cerradoensis sp. nov., a novel isolate from Cerrado soil in Brazil.

An isolate from Cerrado soil, provisionally assigned to the genus Nocardia, was shown to have chemical and morphological properties typical of nocardiae. The strain formed a distinct monophyletic clade in the 16S rDNA tree together with Nocardia africana, Nocardia vaccinii and Nocardia veterana, and showed a unique combination of phenotypic properties that distinguished it from representatives of all recognized species of Nocardia. DNA-DNA relatedness studies indicated that the isolate belongs to a genomic species that is readily distinguished from its nearest neighbours, the type strains of N. africana and N. veterana. The organism is considered to merit species status, and it is proposed that it be designated Nocardia cerradoensis sp. nov., with strain YgT (=CCT 5601T =DSM 44546T) as the type strain.

Brazil↗

Nocardia jejuensis sp. nov., a novel actinomycete isolated from a natural cave on Jeju Island, Republic of Korea.

A novel actinomycete, strain N3-2T, was isolated from a natural cave on Jeju Island, Republic of Korea, using a dilution method and was subjected to polyphasic taxonomy. The almost complete 16S rRNA gene sequence was determined by direct sequencing of the purified PCR product and was compared with those of representatives of the genus Nocardia. It was revealed from the phylogenetic analysis that the organism forms a distinct clade between the Nocardia salmonicida cluster and the Nocardia alba branch within the evolutionary radius occupied by the genus Nocardia of the family Nocardiaceae. The organism showed 16S rRNA gene sequence similarity of 97.4% with its nearest phylogenetic neighbours, namely N. salmonicida and N. alba. The chemotaxonomic properties, such as the principal amino acid of peptidoglycan, predominant menaquinone and polar lipids, supported the classification in the genus Nocardia. The organism was readily differentiated from Nocardia species with validly published names by a broad set of phenotypic properties and its unique phylogenetic position; the name Nocardia jejuensis sp. nov. is proposed, with N3-2T (= JCM 13281T = NRRL B-24430T) as the type strain.

DNA, Ribosomal↗

Differences in the interactions of Nocardia asteroides with macrophage, endothelial, and astrocytoma cell lines.

An in vitro model for studying host cell interactions with Nocardia asteroides was developed. Thus, macrophage cell lines J774A.1 and P388D1, pulmonary artery endothelium cell line CPAE, rat glial tumor cell line C6, and human astrocytoma cell lines CCF-STTG1 and U-373 MG were infected with either log- or stationary-phase cells of N. asteroides GUH-2, and the host cell-nocardia interactions were determined by light microscopy and electron microscopy. Polyclonal antinocardial antibody did not enhance uptake of nocardiae by any of these cell lines; however, log-phase cells of GUH-2 infected a higher percentage of J774A.1 and P388D1 than did stationary-phase organisms. When cells infected with stationary-phase GUH-2 were incubated for 6 h, filaments developed, which indicated that nocardial growth had occurred. In J774A.1 and P388D1, only 31 to 57% of the total stationary-phase coccobacillary cells that were phagocytized formed filaments within 6 h. This indicated that there was some inhibition of growth of the phagocytized nocardiae within these macrophage cell lines; however, the nocardiae grew within the endothelial (> 87% filaments) and astrocytoma (100% filaments) cell lines. Microfilament inhibitor cytochalasin B inhibited uptake of GUH-2 by macrophages and other cell lines, except that there was no effect on uptake of nocardial cells by astrocytoma cell line U-373 MG. Scanning and transmission electron microscopy showed phagocytosis of GUH-2 by the different cell lines. In cytochalasin B-treated cells, nocardiae were shown to penetrate through the cell surface and become internalized in a manner distinct from typical phagocytosis, suggesting that filamentous forms of this organism have a phagocytosis-independent invasion factor. The extent of this cytochalasin-resistant cellular penetration by the nocardiae differed in the different cell lines.

Animals↗

Numerical analysis of the taxonomy of Nocardiae and Rhodococci.

The genera Nocardia and Rhodococcus were clearly differentiated in the present study. Eleven characteristics were shown to be useful for differentiation between these two genera. Nocardia asteroides sunsu stricto previously defined by Tsukamura was divided into two taxa. One contained the type strain and was considered to retain the name Nocardia asteroides in a new sense. Another was named in the present study as Nocardia nova sp. nov. Tsukamura. The type strain of this species is ATCC 33726. The following seven characters were useful for differentiating N. nova from newly defined N. asteroides: 1) arylsulfatase activity after 14 days; 2) catalase activity (semiquantitative); 3) beta-esterase activity; 4) pyrazinamidase activity; 5) utilization of citrate as a sole source of carbon; 6) utilization of 2,3-butylene glycol as a sole carbon source; and 7) resistance to 5-fluorouracil (20 micrograms/ml). The name Nocardia farcinica for Tsukamura's Kyoto-I group should be rejected. This taxon has been named Nocardia paratuberculosis sp. nov. Tsukamura. The type strain is ATCC 23826. Three new species of the genus Rhodococcus were proposed: Rhodococcus aichiensis sp. nov. Tsukamura (type strain, ATCC 33611); Rhodococcus chubuensis sp. nov. Tsukamura (type strain, ATCC 33609); Rhodococcus obuensis sp. nov. Tsukamura (type strain, ATCC 33610).

Actinomycetales↗

Infection with Nocardia species in Queensland. A review of 102 clinical isolates.

OBJECTIVE: To review the species distribution, pathologic significance and disease associations of clinical isolates of Nocardia and related bacteria in Queensland, and to examine the characteristics, treatment and outcome of patients infected with these organisms. DESIGN AND SETTING: A retrospective review of Queensland State Health Laboratory records provided microbiological data for Nocardia isolates referred from other laboratories during the period January 1983 to December 1988. Clinical information was extracted from hospital case notes, or obtained from detailed questionnaires completed by attending physicians. Nocardia isolates were classified as "significant" if specific treatment for nocardiosis was given, or on the basis of autopsy findings. PATIENTS: One hundred and two patients had a Nocardia species or a related organism isolated from clinical specimens during the study period. RESULTS: The 102 isolates included Nocardia asteroides (45), N, brasiliensis (35), N. caviae (5) and N. transvalensis (5). Clinical results were available for 93 patients, of whom 74 (80%) had a significant isolate recovered. Primary pulmonary or disseminated disease occurred in 35 patients, and was caused mainly by N. asteroides. Significant infections of skin and soft tissues, primarily due to N. brasiliensis, were found in 39 patients. Preexisting lung disease and treatment with steroids and immunosuppression were risk factors for pulmonary and disseminated nocardiosis. A history of inoculation in an outdoor setting was frequent in patients with cutaneous disease. Antibiotic regimens that included trimethoprim-sulfamethoxazole or another sulfonamide agent were used to treat the majority of patients with significant infection. Deaths were confined to those with pulmonary and disseminated disease, with a case fatality rate of 40% in that group. CONCLUSION: Infection with Nocardia species appears to be more common than is generally appreciated. The local species distribution and disease spectrum are similar to those described elsewhere. A high index of suspicion for nocardiosis should be maintained in susceptible hosts with pulmonary infiltrates, particularly when there is evidence for metastatic infection, and in patients with superficial infections and a history of outdoor injury.

Adolescent↗

Phenotypic and genotypic profiles of clinical isolates of various Nocardia species to carbapenems and fluoroquinolones.

OBJECTIVES: To establish patterns of the antimicrobial susceptibility of Nocardia species to carbapenems and fluoroquinolones and analysis of phenotypic-genotypic correlations. METHODS: Isolates were identified to the species using 16S rRNA, secA1, or rpoB gene sequencing analysis. The antimicrobial susceptibility testing was performed using the broth microdilution method, and WGS was employed to analyse the presence of resistance genes and/or mutations of Nocardia species against carbapenems and fluoroquinolones. RESULTS: Among 143 Nocardia isolates, N. farcinica (27.27%, 39/143) and N. cyriacigeorgica (25.17%, 36/143) were the most common species, followed by N. abscessus Complex (18.88%, 27/143). The MIC90s of the seven carbapenems were 8 mg/L for doripenem, 8 mg/L for meropenem, 16 mg/L for ertapenem, 16 mg/L for biapenem, 64 mg/L for imipenem, 64 mg/L for faropenem and 128 mg/L for tebipenem, respectively. The susceptibility rates to imipenem were 76.9% and 88.9% for N. farcinica and N. cyriacigeorgica, respectively, but only 14.3% and 0% for N. otitidiscavarium and N. brasiliensis, respectively. Further, 90% of N. brasiliensis and 50% of N. otitidiscaviarum isolates were susceptible and intermediate to meropenem. WGS identified blaFAR-1 gene in N. farcinica and blaAST-1 gene in N. cyriacigeorgica, respectively. The MIC90s of the four fluoroquinolones were 1 mg/L for sitafloxacin, 4 mg/L for nemonoxacin, 4 mg/L for moxifloxacin and 16 mg/L for ciprofloxacin, respectively. The susceptibility rate of Nocardia species to ciprofloxacin was low except for N. farcinica. The resistance to fluoroquinolones arise from mutations in the gyrA gene. CONCLUSIONS: Nocardia spp. exhibited varying patterns of susceptibility to carbapenems and fluoroquinolones respectively. Importantly, different Nocardia spp. exhibited different patterns of susceptibility to carbapenems and fluoroquinolones, respectively.

Nocardia↗

Nocardia concava sp. nov., isolated from Japanese patients.

Two actinomycete strains, IFM 0354(T) and IFM 0576, isolated from Japanese patients, were found to have morphological, biochemical and chemotaxonomic properties consistent with their classification in the genus Nocardia. The strains resembled Nocardia otitidiscaviarum and Nocardia uniformis in their phenotypic properties. Comparative 16S rRNA gene sequence analysis showed that the strains are closely related to Nocardia seriolae. DNA-DNA relatedness values and phenotypic differences from N. seriolae indicated that the strains belong to a novel species of Nocardia, for which the name Nocardia concava sp. nov. is proposed. The type strain is IFM 0354(T) (=NBRC 100430(T)=JCM 12351(T)=DSM 44804(T)).

Bacterial Typing Techniques↗

Rapid identification of clinically relevant Nocardia species using real-time PCR with SYBR Green and melting-curve analysis.

The objective of this study was to develop and evaluate a rapid new method of identifying clinically relevant Nocardia species. DNA extracted from different Nocardia strains was used in a real-time PCR assay with SYBR Green and melting-curve analysis to identify Nocardia species. Ten control strains and four bacterial strains of closely related genera were employed, and samples from 28 patients were used. All Nocardia strains were identified correctly, and there was no cross-reaction with strains from genera closely related to Nocardia. The sensitivity and specificity of the method were 90 and 100 %, respectively. This method can be used to rapidly detect Nocardia species in culture, without cross-reaction with other closely related genera.

Humans↗

Nocardia aobensis Sp. Nov., isolated from patients in Japan.

Five clinical isolates, strains IFM 0137, 0372(T), 0496, 0556, and 0952, were provisionally assigned to the genus Nocardia based on morphological criteria. Nearly complete 16S rDNA sequences were determined for these strains. These data showed that they are most similar to that of Nocardia africana, Nocardia cerradoensis and Nocardia veterana. However, DNA-DNA relatedness data showed that the five strains were of a single species and were distinguishable from N. africana, N. cerradoensis and N. veterana. Therefore, these strains represent a new species within the genus Nocardia. The designation of these five strains is Nocardia aobensis sp. nov. The type strain is IFM 0372(T) (=NBRC 100429(T)=JCM 12352(T)=DSM 44805(T)).

Bacterial Typing Techniques↗

Identification of nocardia species by restriction endonuclease analysis of an amplified portion of the 16S rRNA gene.

Identification of clinical isolates of Nocardia to the species level is important for defining the spectrum of disease produced by each species and for predicting antimicrobial susceptibility. We evaluated the usefulness of PCR amplification of a portion of the Nocardia 16S rRNA gene and subsequent restriction endonuclease analysis (REA) for species identification. Unique restriction fragment length polymorphism (RFLP) patterns were found for Nocardia sp. type strains (except for the N. asteroides type strain) and representative isolates of the drug pattern types of Nocardia asteroides (except for N. asteroides drug pattern type IV, which gave inconsistent amplification). A variant RFLP pattern for Nocardia nova was also observed. Twenty-eight clinical isolates were evaluated both by traditional biochemical identification and by amplification and REA of portions of the 16S rRNA gene and the 65-kDa heat shock protein (HSP) gene. There was complete agreement among the three methods on identification of 24 of these isolates. One isolate gave a 16S rRNA RFLP pattern consistent with the biochemical identification but was not identifiable by its HSP gene RFLP patterns. Three isolates gave 16S rRNA RFLP patterns which were inconsistent with the identification obtained by both biochemical tests and HSP gene RFLP; sequence analysis suggested that two of these isolates may belong to undefined species. The PCR and REA technique described appears useful both for the identification of clinical isolates of Nocardia and for the detection of new or unusual species.

Bacterial Proteins↗

Nocardia africana sp. nov., a new pathogen isolated from patients with pulmonary infections.

Eight actinomycete strains, isolated from 8 out of 400 sputum samples examined, taken from patients with pulmonary diseases at the Chest Unit of Khartoum Teaching Hospital in the Sudan, were provisionally assigned to the genus Nocardia according to morphological criteria. These isolates were studied further in order to establish their taxonomic status. They were found to have morphological and chemical properties typical of nocardiae and formed a monophyletic clade in the 16S ribosomal DNA tree together with Nocardia vaccinii. The strains showed a unique pattern of phenotypic properties that distinguished them from representatives of recognized Nocardia species, including Nocardia vaccinii. The strains were considered to merit species status and were designated Nocardia africana sp. nov. The findings of the present study are consistent with the view that pulmonary nocardiosis may occur in a substantial proportion of patients who exhibit chronic lung diseases in African countries. It is important, therefore, that clinicians in such countries consider this condition, especially when patients with respiratory infections fail to respond to antitubercular therapy.

Africa↗

Nocardia veterana isolated from ascitic fluid of a patient with human immunodeficiency virus infection.

Nocardia veterana is a recently characterized species within the genus Nocardia, and only three human clinical isolates have been reported for this species. We describe a case of ascitic fluid infection in an immunocompromised patient due to N. veterana. To our knowledge, this is the first report of a Nocardia sp. strain from ascitic fluid and the fourth report of N. veterana isolated from human samples. Chemotaxonomic methods showed the strain to belong to the genus Nocardia, and identification to the species level was done by 16S ribosomal DNA gene sequencing. The antibiotic susceptibility profile of N. veterana is reported here for the second time. The strain was deposited in the Collection of the Pasteur Institute and in the Culture Collection of the University of Göteborg (CIP 107497 and CCUG 46576). The corresponding 16S ribosomal DNA gene sequence is available from the GenBank database under accession number AY149599. A phylogenetic analysis was conducted and showed that N. veterana was most closely related to the recently characterized species Nocardia africana rather than to Nocardia vaccinii, as previously reported.

AIDS-Related Opportunistic Infections↗

[Nocardia infections: clinical and biological aspects].

The nocardiosis is an infection caused by a bacterial pathogen agent, Nocardia, belonging to the Actinomycetales order. They are Gram-positive, strictly aerobic bacteria. Members of the genus Nocardia are ubiquitous. They are frequently isolated from soil, water, air dusts. The mode of contamination occurs by inhalation or by cutaneous or ocular traumatic lesion. Clinically, nocardiosis is essentially characterized by pulmonary diseases. Others secondary localizations are described, such as in the central nervous system. Nocardia can be responsible for important cutaneous, subcutaneous and lymphocutaneous manifestations. In the same way, some extrapulmonary diseases and spread nocardiosis are more rarely observed. Several factors seem to favour the development of Nocardia. The immunocompromised patients, particularly those with organ transplant and the patients treated with immunosuppressor treatments, offer strong predispositions to this opportunistic disease. The nocardiosis is nevertheless observed in healthy persons. In front of polymorphic and specific-less clinical manifestations, large phenotypic heterogeneity, and resistance profiles to specific antibiotics, a correct diagnosis for Nocardia species is necessary to apply an adequate treatment. The techniques of identification based on the chemotaxonomic analysis and the susceptibility to different inhibitors are efficient for the identification of genus and species. However, because of the slow growth rate of Nocardia, the reading of these tests can require several weeks of incubation. With the intention of the rapid identification of genus and species, the molecular techniques (PCR-RFLP) seem to be efficient. The technique of RAPD allows an efficient molecular typing, which will give a better knowledge concerning transmission, ecological niches and epidemic reservoirs.

Central Nervous System Bacterial Infections↗