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PubMed2005

Lack of evidence for Nocardia asteroides in brain specimens from Lewy body-containing disorders.

Previous studies have suggested that Nocardia asteroides may play a role in the pathogenesis of Parkinson's disease (PD), including the production of Lewy bodies, the inclusion bodies present in this disorder. This study explored the possible connection between Nocardia and two Lewy body-containing disorders, PD and dementia with Lewy bodies (DLB). Substantia nigra specimens from individuals with PD, DLB, other neurodegenerative disorders, and normal subjects were evaluated for nocardial infection by in situ hybridization, PCR, and Gram staining. Brain specimens from a cynomolgus monkey experimentally infected with N. asteroides for 48 h served as the controls for in situ hybridization and Gram staining, and a nocardial pellet was the PCR control. The organism was detected by in situ hybridization and Gram stain in the experimentally infected monkey brain, and by PCR from the nocardial pellet. However, in situ hybridization reactivity was detected in only three of the 125 human brain specimens (2.4%; one case each of PD, DLB, and Alzheimer's disease), and none of the specimens was positive for Nocardia by PCR or Gram staining. These findings do not support an association of Nocardia with Lewy body-containing disorders.

Adult↗
PubMed1990

Characterization of Nocardia, Rhodococcus and Gordona species by in vitro susceptibility testing.

Representative strains of Gordona, Nocardia and Rhodococcus were tested against 26 antimicrobial agents using the disc diffusion method. A distinct susceptibility profile was noted for most species. Nocardia asteroides, N. brasiliensis, N. otitidiscaviarum, N. transvalensis and N. vaccinii were rarely susceptible to the antibiotics tested; N. brevicatena and N. farcinica varied in their susceptibility depending on the strain tested. Only the antibiotic amikacin was active against all nocardiae. In contrast, the Gordona and Rhodococcus strains showed considerable susceptibility, in particular to the beta-lactam antibiotics. Amoxicillin + clavulanic acid and gentamicin were active against all of the gordonae and rhodococci. The differences in susceptibility patterns may prove useful in characterizing the genera Gordona, Nocardia and Rhodococcus and in separating species within these taxa.

Anti-Bacterial Agents↗
PubMed2002

Nocardia infection in chronic granulomatous disease.

To determine the clinical characteristics and outcome of Nocardia infection in patients with chronic granulomatous disease (CGD), we reviewed data on 28 episodes of Nocardia infection in 23 patients with CGD. All episodes involved pulmonary infection. The frequency of disseminated nocardiosis was 25% for the case series overall, but it was 56% among episodes in patients receiving neither interferon-gamma (IFN-gamma) nor sulfonamide prophylaxis. Patients receiving prophylaxis with IFN-gamma and/or a sulfonamide were significantly less likely to have disseminated nocardiosis than were patients not receiving these medications, and no patient receiving both medications developed disseminated nocardiosis. One-third of the patients had concomitant fungal infections, and 2 patients had concomitant Legionella infections. The majority of patients were successfully treated with a sulfonamide-containing regimen, even though some patients had developed Nocardia infection while receiving sulfonamide prophylaxis. Nocardia infections in patients with CGD are not usually fatal if treated properly, and prophylaxis with IFN-gamma and a sulfonamide may protect against dissemination.

Adolescent↗
PubMed1998

Nocardia abscess of the choroid plexus: clinical and pathological case report.

OBJECTIVE: Cerebral Nocardia abscesses are rare, accounting for approximately 1 to 2% of all cerebral abscesses. Prompt aggressive surgical treatment involving craniotomy and excision of these lesions has been advocated by many authors, because these lesions have significantly higher morbidity and mortality rates than do most other cerebral abscesses. We report an atypical presentation of cerebral nocardiosis localized to the choroid plexus of the lateral ventricle. CLINICAL PRESENTATION: A 56-year-old man presented with a 3-week history of fever, cough, and progressive headache and an ensuing 3-day history of progressive lethargy, confusion, and gait ataxia. Radiographic studies demonstrated a loculated contrast-enhancing left lateral ventricular lesion with significant perilesional parenchymal edema that was thought preoperatively to be a neoplasm. INTERVENTION: The patient underwent a craniotomy for resection of the lesion. Intraoperatively, a reddish gray lesion with purulent exudate was encountered within the left lateral ventricle intimately adherent to the choroid plexus as well as to the ependyma and subependymal veins. A frozen section demonstrated an organizing abscess wall. The lesion was resected in its entirety, and multiple cultures were sent for analysis. CONCLUSION: Microbiology cultures grew Nocardia asteroides. A course of intravenous antibiotics was started, which included trimethoprim-sulfamethoxazole, amikacin, and ceftriaxone. Two weeks after surgery, at the time of discharge, the patient's neurological status had improved considerably. Although Nocardia abscesses have been documented to occur throughout the central nervous system, the presentation of a lesion confined to the choroid plexus of the lateral ventricle with significant parenchymal edema is unusual and demonstrates that Nocardia abscesses must be considered in the differential diagnosis of a contrast-enhancing intraventricular mass lesion involving the choroid plexus.

Brain Abscess↗
PubMed2000

Nocardia abscessus sp. nov.

Chemotaxonomic and 16S rRNA gene sequence analyses of four bacterial strains isolated from clinical material clearly demonstrated that these bacteria belong to the genus Nocardia. DNA-DNA hybridization data as well as the physiological characteristics of the isolates indicated that they are closely related and belong to a single species that differs from previously described members of the genus. The name Nocardia abscessus sp. nov. is proposed for these organisms represented by strain IMMIB D-1592T (= DSM 44432T). Strain IMMIB D-1592T exhibits 56.8 and 60.0% DNA-DNA relatedness to Nocardia asteroides ATCC 19247T and Nocardia paucivorans DSM 44386T, respectively.

Adult↗
PubMed2001

Nocardia veterana sp. nov., isolated from human bronchial lavage.

A nocardioform bacterium was isolated from the bronchoscopic lavage of a 78-year-old man with a past history of tuberculous pleurisy, who presented with bilateral upper lobe lesions at Austin and Repatriation Medical Centre, Heidelberg, Australia. The strain was aerobic, Gram-positive, produced beige substrate mycelium and scant white aerial mycelium. It showed chemotaxonomic markers which were consistent with the classification of Nocardia: i.e. meso-diaminopimelic acid, N-glycolylmuramic acid, arabinose and galactose as diagnostic sugars; phospholipids phosphatidylinositol mannosides, phosphatidylinositol, phosphatidylethanolamine and diphosphatidylglycerol; a menaquinone with a cyclic isoprene side chain, MK-8(H4cycl.); a fatty acid pattern composed of unbranched saturated and monounsaturated fatty acids with a considerable amount of tuberculostearic acid; and mycolic acids composed of 54-62 carbon atoms with three principal mycolic acids which were mono- and polyunsaturated, showing a chain length C56, C58 and C60 and accounting for over 70% of the entire pattern. The 16S rDNA sequence showed the highest similarity to the type strain of Nocardia vaccinii; the DNA-DNA similarity of the two strains was 31%. These data, together with distinct physiological traits and molecular biological analyses, as well as chemotaxonomic results, led to the conclusion that the novel isolate represents a new species within the genus Nocardia for which the name Nocardia veterana sp. nov. is proposed. The type strain is M157222T (DSM 44445T = NRRL B-24136T).

Aged↗
PubMed2001

Nocardia ignorata sp. nov.

An isolate that was received during a mycobacterial quality control test and which was thought to be a Mycobacterium species was subjected to a polyphasic taxonomic study after mycolic acid analysis showed that it possessed mycolates characteristic of the nocardiae. Further chemotaxonomic and 16S rRNA gene sequence analyses of this isolate demonstrated that it belongs to the genus Nocardia. 16S rRNA gene sequence data, DNA-DNA hybridization studies and the biochemical characteristics of the isolate indicate that it belongs to a novel species that differs from previously described members of the genus Nocardia. The name Nocardia ignorata sp. nov. is proposed for this isolate with the type strain IMMIB R-1434T (= DSM 44496T = NRRL B-24141T).

DNA, Ribosomal↗
PubMed2006

Nocardia harenae sp. nov., an actinomycete isolated from beach sand.

A novel actinomycete, strain WS-26(T), was isolated from beach sand on the coast of Jeju Island, Republic of Korea, and subjected to a polyphasic taxonomic characterization. The organism produced reddish orange-coloured substrate mycelium and white aerial mycelium, both of which fragmented into rod-shaped elements. A neighbour-joining tree, based on 16S rRNA gene sequence studies, revealed that the isolate formed a distinct branch at the base of a cluster that included Nocardia carnea, N. flavorosea, N. pigrifrangens, N. sienata and N. testacea. This branching pattern was also found in a tree constructed using the maximum-likelihood method, but was not supported by the maximum-parsimony method. Levels of 16S rRNA gene sequence similarity between the isolate and its phylogenetic neighbours ranged from 96.5 to 97.8 %. Chemotaxonomic properties, such as the principal amino acid of peptidoglycan, predominant menaquinone and polar lipids, supported its assignment to the genus Nocardia. On the basis of phenotypic and phylogenetic data, the organism was different from Nocardia species with validly published names. The name Nocardia harenae sp. nov. is proposed, with WS-26(T) (=KCCM 42317(T)=NRRL B-24459(T)) as the type strain.

Bacterial Typing Techniques↗
PubMed1994

Transfer of Nocardia amarae Lechevalier and Lechevalier 1974 to the genus Gordona as Gordona amarae comb. nov.

The taxonomic status of Nocardia amarae strains was examined using chemical, microbiological and nucleic acid sequencing methods. It was evident from the results of this and previous studies that Nocardia amarae has properties that are at variance with its classification in the genus Nocardia but consistent with its transfer to the genus Gordona. It is proposed that Nocardia amarae Lechevalier and Lechevalier 1974 be transferred to the genus Gordona as Gordona amarae comb. nov.

Actinomycetales↗
PubMed1984

Disseminated cutaneous Nocardia brasiliensis infection.

Nocardia brasiliensis is an uncommon pathogen in the United States despite its presence in soil samples. It is most frequently seen in patients who are immunosuppressed, but can occur in the immunocompetent host also. Usually in a healthy host the infection is localized to one area of the body. Disseminated infection was diagnosed in a healthy child who recovered fully after initiation of oral sulfonamide therapy. We suspect that the source was soil to which the patient was exposed during a family camping trip. Nocardia asteroides is the most frequent cause of nocardial infections in North America. These infections are unusual in healthy individuals, occurring more often in immunologically compromised hosts. They are usually acquired by a pulmonary route and may disseminate widely, most commonly to the central nervous system and skin. Nocardia brasiliensis, when it does occur in the United States, usually causes a chronic cutaneous and subcutaneous infection manifesting itself as mycetoma, abscess, or ulcer. We saw acute suppurative lymphocutaneous Nocardia brasiliensis infection in an otherwise healthy 2-year-old boy.

Child, Preschool↗
PubMed1991

Cloning, nucleotide sequence, and transcriptional analysis of the NAD(P)-dependent cholesterol dehydrogenase gene from a Nocardia sp. and its hyperexpression in Streptomyces spp.

NAD(P)-dependent cholesterol dehydrogenases [NAD(P)-CDH], which allow easier quantification of cholesterol by means of directly measuring the A340 of NAD(P)H, are useful for clinical purposes. The amino acid sequences of the NH2 terminus and the fragments obtained by CNBr decomposition of the NAD(P)-CDH from a Nocardia sp. were determined for preparation of synthetic oligonucleotides as hybridization probes. A 4.4-kbp BamHI fragment hybridizing to these probes was cloned on pUC19 in Escherichia coli. The nucleotide sequence together with the determined amino acid sequences revealed that this enzyme consists of 364 amino acids (Mr, 39,792) and contains an NAD(P)-binding consensus sequence at its NH2-terminal portion. High-resolution S1 nuclease mapping suggested that in NAD(P)-CDH of both Nocardia and Streptomyces spp. transcription initiates at the adenine residue, which is the first position of the translational initiation triplet (AUG) of this protein. The S1 mapping experiments also showed that cholesterol-dependent regulation in the Nocardia sp. occurred at the level of transcription. In Streptomyces lividans containing the cloned fragment, however, this promoter was expressed constitutively. DNA manipulation of the cloned gene in E. coli, including the generation of a ribosome-binding sequence at an appropriate position by oligonucleotide-directed mutagenesis, led to production of this protein in a very large amount but in the enzymatically inactive form of inclusion bodies. On the other hand, a Streptomyces host-vector system was successfully used for producing 40 times as much enzymatically active NAD(P)-CDH as that produced by the original Nocardia sp.

3-Hydroxysteroid Dehydrogenases↗
PubMed2003

GTP cyclohydrolase I: purification, characterization, and effects of inhibition on nitric oxide synthase in nocardia species.

GTP cyclohydrolase I (GTPCH) catalyzes the first step in pteridine biosynthesis in Nocardia sp. strain NRRL 5646. This enzyme is important in the biosynthesis of tetrahydrobiopterin (BH4), a reducing cofactor required for nitric oxide synthase (NOS) and other enzyme systems in this organism. GTPCH was purified more than 5,000-fold to apparent homogeneity by a combination of ammonium sulfate fractionation, GTP-agarose, DEAE Sepharose, and Ultragel AcA 34 chromatography. The purified enzyme gave a single band for a protein estimated to be 32 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass of the native enzyme was estimated to be 253 kDa by gel filtration, indicating that the active enzyme is a homo-octamer. The enzyme follows Michaelis-Menten kinetics, with a Km for GTP of 6.5 micromoles. Nocardia GTPCH possessed a unique N-terminal amino acid sequence. The pH and temperature optima for the enzyme were 7.8 and 56 degrees C, respectively. The enzyme was heat stable and slightly activated by potassium ion but was inhibited by calcium, copper, zinc, and mercury, but not magnesium. BH4 inhibited enzyme activity by 25% at a concentration of 100 micromoles. 2,4-Diamino-6-hydroxypyrimidine (DAHP) appeared to competitively inhibit the enzyme, with a Ki of 0.23 mM. With Nocardia cultures, DAHP decreased medium levels of NO2- plus NO3-. Results suggest that in Nocardia cells, NOS synthesis of nitric oxide is indirectly decreased by reducing the biosynthesis of an essential reducing cofactor, BH4.

Biopterins↗
PubMed1996

A rapid and gentle method for isolation of genomic DNA from pathogenic Nocardia spp.

The lack of simple and efficient methods for extraction of DNA from Nocardia spp. has hampered molecular manipulation of the DNA for diagnostic purposes. In the present study, a method for the rapid extraction of undegraded genomic nocardial DNA was established. Briefly, 14 pathogenic Nocardia strains were grown at 37 degrees C for 3 to 5 days in Sauton broth containing 0.05% Tween 80. Subsequently, the cultures were treated for 48 h with 1.2 mg of cycloserine per ml (final concentration). Cells were then harvested by centrifugation and treated with a lysis solution containing 3 mg of lysozyme per ml. This was followed by the addition of proteinase K and sodium dodecyl sulfate to final concentrations of 0.2 mg/ml and 0.5%, respectively, and incubation for 1 h at 50 degrees C. DNA was precipitated with isopropanol after phenol-chloroform-isoamyl alcohol extractions and RNase treated before being quantitated and analyzed by agarose gel electrophoresis. The average undegraded DNA yields obtained were 101 micrograms for Nocardia brasiliensis and 121 micrograms for N. asteroides. This DNA was suitable for restriction endonuclease digestion and PCR amplification, which are methods being applied to the characterization and diagnosis of slowly growing organisms such as Nocardia spp.

DNA Restriction Enzymes↗
PubMed1979

B-lymphocyte activation with an extract of Nocardia brasiliensis.

An extract from the pathogenic actinomycete Nocardia brasiliensis was mitogenic for murine lymphocytes. This deoxyribonucleic acid-synthetic response of whole spleen cells peaked after 48 h in culture at concentrations of Nocardia extract ranging from 10 to 200 micrograms/ml. The extract appeared to be a mitogen for B lymphocytes since cultures of spleen cells from congenitally athymic nude (nu/nu) mice and of antithymocyte serum plus complement-treated spleen cells from conventional (+/+) mice responded as well as untreated spleen cells from normal +/+ mice. Furthermore, thymocytes did not respond mitogenically to the extract. Mitogenic responses were stimulated in spleen cells from H-2(a), H-2(b), H-2(d), and H-2(k) mice, including lipopolysaccharide-nonresponder C3H/HeJ mice. This Nocardia extract also stimulated polyclonal B-cell activation to the hapten trinitrophenyl, serum protein human gamma globulin, and several mammalian erythrocytes in cultures of cells from both euthymic and nude mice. Additionally, the requirement for helper T cells in the primary in vitro immune response to sheep erythrocytes could be circumvented by the addition of this Nocardia extract. These results indicate that an extract from the pathogen N. brasiliensis can nonspecifically activate murine B lymphocytes and raise the possibility that polyclonal activation of B lymphocytes may contribute to the pathogenesis of nocardiosis.

Animals↗
PubMed1980

Comparative studies of antigen 21 in Mycobacterium and Nocardia species: possible taxonomic relationships with Mycobacterium leprae.

Studies of Mycobacterium leprae, Mycobacterium tuberculosis and Nocardia caviae in comparison with each other and with other Mycobacterium and Nocardia species were performed on the basis of antigen 21 intramolecular heterogeneity. Three different antisera were used: rabbit anti-Mycobacterium smegmatis antiserum, rabbit anti-Nocardia asteroides antiserum, and a lepromatous serum pool. With reference to each of the three antiserum sources used the strains were ranked in an order of relatedness or sharing of determinants. The three antisera showed distinctly different antigen 21 antibody specificities reflecting the species origin of the immunogen. The present investigations confirmed that antigen 21 of N. caviae shares determinants with antigens from Mycobacterium strains which were not present in corresponding antigens of all other Nocardia strains tested. M. tuberculosis, as judged by antigen 21 analysis, occupies a position separate from both the slow-growing and the fast-growing mycobacterial clusters in accordance with accepted taxonomic relationships. An interesting possibility of establishing a position for M. leprae in relation to other mycobacterial species was apparent. The order of relatedness among the strains studied went from M. leprae to M. tuberculosis to N. caviae to Mycobacterium avium to Mycobacterium fortuitum, the last two being representatives of the slow-growing and fast-growing mycobacteria. It can therefore be concluded that evidence from antigen 21 analysis indicates that M. leprae is more closely related to M. tuberculosis than to the other strains investigated.

Antigens, Bacterial↗
PubMed1973

Serological study of nocardia by using mycobacterial precipitation reference systems.

Forty-six nocardia strains and ten strains labeled "Mycobacterium" were analyzed with the comparative immunodiffusion technique, employing as reference material nine mycobacterial precipitation systems, each one representing a different species. Three precipitinogens were common for many of the tested strains and the mycobacterial reference strains. The serological analyses indicated the possibility of differentiating two groups within Nocardia depending on the characteristics of one of the common precipitinogens. Five of eight tested Nocardia farcinica strains showed a serologically close relationship with the mycobacteria and might be regarded as belonging to this genus. The "M." pellegrino and "M." rhodochrous strains, however, did not react with the antimycobacterial sera to a higher degree than did the tested nocardia strains.

Animals↗
PubMed2000

Cyclic guanosine-3',5'-monophosphate and biopteridine biosynthesis in Nocardia sp.

Nocardia sp. strain NRRL 5646 contains a nitric oxide synthase (NOS) enzyme system capable of generating nitric oxide (NO) from arginine and arginine-containing peptides. To explain possible roles of the NOS system in this bacterium, guanylate cyclase (GC) and tetrahydrobiopterin (H(4)B) biosynthetic enzymes were identified in cell extracts and in culture media. Cell extracts contained GC activity, as measured by the conversion of GTP to cyclic guanosine-3',5'-monophosphate (cGMP) at 9.56 pmol of cGMP h(-1) mg of protein(-1). Concentrations of extracellular cGMP in culture media were significantly increased, from average control levels of 45 pmol cGMP liter(-1) to a maximum of 315 pmol liter(-1), in response to additions of GTP, L-arginine, H(4)B, and sodium nitroprusside to growing Nocardia cultures. On the other hand, the NOS inhibitor N(G)-nitro-L-arginine and the GC inhibitor 1H-[1,2, 4]oxadiazole[4,3-a]quinoxalin-1-one both dramatically decreased extracellular cGMP levels. Activities for GTP-cyclohydrase-1, 6-pyruvoyltetrahydropterin synthase and sepiapterin reductase, enzymes essential for H(4)B biosynthesis, were present in Nocardia culture extracts at 77.5 pmol of neopterin and 45.8 pmol of biopterin h(-1) mg of protein(-1), respectively. In Nocardia spp., as in mammals, GTP is a key intermediate in H(4)B biosynthesis, and GTP is converted to cGMP by a GC enzyme system that is activated by NO.

Alcohol Oxidoreductases↗
PubMed1997

Comparison of agar dilution, broth microdilution, disk diffusion, E-test, and BACTEC radiometric methods for antimicrobial susceptibility testing of clinical isolates of the Nocardia asteroides complex.

An evaluation was undertaken to determine the optimal method for the in vitro susceptibility testing of 26 Nocardia asteroides complex isolates to the following antimicrobial agents: amikacin, ampicillin, amoxicillin-clavulanate, ceftriaxone, ciprofloxacin, erythromycin, imipenem, minocycline, and trimethoprim-sulfamethoxazole. Five testing methods were studied including the agar dilution, broth microdilution, and disk diffusion methods, the epsilometer test (E-test), and the BACTEC radiometric method. Results for each antimicrobial agent and each testing method were interpreted as indicating susceptibility, intermediate susceptibility, or resistance according to current guidelines of the National Committee for Clinical Laboratory Standards (NCCLS) for bacteria that grow aerobically and were then compared to a "gold standard" susceptibility test result. The gold standard result for each Nocardia isolate was established by a consensus of the results of the majority of testing methods used in the study. When the results were combined for all antimicrobial agents tested against all Nocardia isolates by all methods, the BACTEC radiometric method produced the highest level of agreement (97.9%) with the consensus results and had the fewest very major (n = 1), major (n = 2), and minor (n = 2) errors. In contrast, the results of the agar dilution method were in least agreement (93.2%) with the consensus results, and this method also produced the most very major (n = 8), major (n = 4), and, along with the disk diffusion method, minor (n = 6) errors. For all test methods, interpretive errors were most frequent when testing ampicillin or amoxicillin-clavulanate. Moreover, for all Nocardia nova isolates tested, ampicillin susceptibility results by any of the testing methods were not in agreement with the results of testing for beta-lactamase by the nitrocefin (Cefinase) disk method. We conclude that among the methods evaluated, the BACTEC radiometric method appeared to be the best for determining the in vitro susceptibilities of members of the N. asteroides complex to a panel of nine antimicrobial agents. However, none of the test methods, including the BACTEC method, accurately predicted the ampicillin resistance of the N. nova isolates tested, all of which produced beta-lactamase. Presuming that this beta-lactamase hydrolyzes ampicillin, this disparity may relate to the NCCLS breakpoints that were used, which may require modification for this antimicrobial agent when tested against N. nova isolates.

Drug Resistance, Microbial↗