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PubMed2003

Rapid identification of Rhodococcus equi by a PCR assay targeting the choE gene.

The actinomycete Rhodococcus equi is an important pathogen of horses and an emerging opportunistic pathogen of humans. Identification of R. equi by classical bacteriological techniques is sometimes difficult, and misclassification of an isolate is not uncommon. We report here on a specific PCR assay for the rapid and reliable identification of R. equi. It is based on the amplification of a fragment of the choE gene encoding cholesterol oxidase. The choE-based PCR was assessed by using a panel of strains comprising 132 isolates from different sources and of different geographical origins, all initially identified biochemically as R. equi, and 30 isolates of representative non-R. equi actinomycete species, including cholesterol oxidase producers. The expected 959-bp amplicon was observed only with R. equi isolates, as confirmed by sequencing of a variable region of the 16S RNA gene from a random sample of 20 PCR-positive isolates. All R. equi isolates gave a positive choE-based PCR result, which correlated with a high degree of conservation of the choE gene. Three of the 132 strains originally identified as R. equi were negative for the choE gene, and subsequent analysis of their 16S RNA gene sequences confirmed that they belonged to other bacterial species (Dietzia maris, Mycobacterium peregrinum, and Staphylococcus epidermidis). All non-R. equi isolates were negative by the choE-based PCR. ATCC 21387, the only known isolate of Brevibacterium sterolicum, gave a 959-bp amplicon whose DNA sequence was virtually identical to that of R. equi choE. Comparison of the 16S RNA genes indicated that ATCC 21387 should be considered an R. equi isolate.

Actinomycetales Infections↗
PubMed2003

Virulence of Rhodococcus equi isolated from cats and dogs.

Nine cat isolates and nine dog isolates of Rhodococcus equi from clinical material were investigated for the presence of the virulence-associated antigens (VapA and VapB) and virulence plasmids. Five of the cat isolates and one dog isolate were VapA positive and contained an 85-kb type I or an 87-kb type I plasmid. The remaining 12 isolates were avirulent R. equi strains and contained no virulence plasmids.

Animals↗
PubMed2005

Pyogenic liver abscess due to Rhodococcus equi in an immunocompetent host.

A case of pyogenic liver abscess (PLA) due to Rhodococcus equi in an immunocompetent individual was successfully treated by combining surgery and antibiotics. The R. equi-targeted antimicrobial agents erythromycin and rifampin were used only after surgical resection of the lesion and identification of the infective organism.

Actinomycetales Infections↗
PubMed2005

Characterization of virulence plasmids and serotyping of rhodococcus equi isolates from submaxillary lymph nodes of pigs in Hungary.

The plasmid types and serotypes of 164 Rhodococcus equi strains obtained from submaxillary lymph nodes of swine from different piggeries in 28 villages and towns located throughout the country were examined. The strains were tested by PCR for the presence of 15- to 17-kDa virulence-associated protein antigen (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphism characteristics. None of the 164 isolates contained the vapA gene, and 44 (26.8%) isolates were positive for the vapB gene, showing a product of the expected 827-bp size in the PCR amplification. The 44 isolates of intermediate virulence contained virulence plasmids that were identified as types 1 (3 isolates), 4 (1 isolate), 5 (36 isolates), 6 (1 isolate), and 7 (2 isolates) and as a new variant (1 isolate). On the basis of restriction digestion patterns of plasmid DNAs, we tentatively designated the variant as type 17. Use of the serotyping method of Prescott showed that 110 (67.1%) out of the 164 isolates were typeable and that serotype 2 predominated (83 isolates [50.6%]), followed by serotype 1 (26 strains [15.9%]). Only one isolate belonged to serotype 3. A total of 54 (32.9%) isolates were untypeable in Prescott's system. The prevalence of R. equi strains of intermediate virulence among the isolates that came from the submaxillary lymph nodes of swine in Hungary was lower than that seen with isolates obtained elsewhere.

Animals↗
PubMed2026

Complete genome sequence of Rhodococcus qingshengii strain A3-8.

A chemostat culture was constructed with phenol and forest soil as an inoculum. We report the complete genome sequence of Rhodococcus qingshengii strain A3-8, which was isolated from the culture. The genome consists of a chromosome (6,436,695 bp) and a linear plasmid pA38 (257,365 bp).

Rhodococcus↗
PubMed2006

Specificity of catechol ortho-cleavage during para-toluate degradation by Rhodococcus opacus 1cp.

Degradation of para-toluate by Rhodococcus opacus 1cp was investigated. Activities of the key enzymes of this process, catechol 1,2-dioxygenase and muconate cycloisomerase, are detected in this microorganism. Growth on p-toluate was accompanied by induction of two catechol 1,2-dioxygenases. The substrate specificity and physicochemical properties of one enzyme are identical to those of chlorocatechol 1,2-dioxygenase; induction of the latter enzyme was observed during R. opacus 1cp growth on 4-chlorophenol. The other enzyme isolated from the biomass grown on p-toluate exhibited lower rate of chlorinated substrate cleavage compared to the catechol substrate. However, this enzyme is not identical to the catechol 1,2-dioxygenase cloned in this strain within the benzoate catabolism operon. This supports the hypothesis on the existence of multiple forms of dioxygenases as adaptive reactions of microorganisms in response to environmental stress.

Adaptation, Physiological↗
PubMed1987

Non-pulmonary Rhodococcus equi infections in patients with acquired immune deficiency syndrome (AIDS).

Rhodococcus equi, formerly known as Corynebacterium equi, was isolated repeatedly from the blood of two patients with the acquired immune deficiency syndrome (AIDS). Neither of the patients had pneumonia while they were bacteraemic, whereas pneumonia has been present in all previously reported cases of human infection with R equi. One of our patients had diarrhoea and the organism was isolated from a stool culture; the other patient had a large granulomatous soft tissue mass in his pelvis caused by R equi. Both isolates were resistant to penicillin and one produced a beta-lactamase. Both patients were treated with vancomycin but only one recovered.

Acquired Immunodeficiency Syndrome↗
PubMed1999

Infection by Rhodococcus equi in a patient with AIDS: histological appearance mimicking Whipple's disease and Mycobacterium avium-intracellulare infection.

Rhodococcus equi pneumonia with systemic dissemination is being reported increasingly in immunocompromised patients. This is the first case report of disseminated R equi infection with biopsy documented involvement of the large intestine. The patient was a 46 year old male with AIDS who was diagnosed with cavitating pneumonia involving the left lower lobe. R equi was isolated in culture from the blood and lung biopsies. Subsequently, the patient developed anaemia, diarrhoea, and occult blood in the stool. Colonoscopy revealed several colonic polyps. Histological examination of the colon biopsies showed extensive submucosal histiocytic infiltration with numerous Gram positive coccobacilli and PAS positive material in the histiocytes. Electron microscopy showed variably shaped intrahistiocytic organisms which were morphologically consistent with R equi in the specimen. Disseminated R equi infection may involve the lower gastrointestinal tract and produce inflammatory polyps with foamy macrophages which histologically resemble those seen in Whipple's disease and Mycobacterium avium-intracellulare infection.

AIDS-Related Opportunistic Infections↗
PubMed1986

Review of Corynebacterium (Rhodococcus) equi lung abscesses in foals: pathogenesis, diagnosis and treatment.

Corynebacterium (Rhodococcus) equi is becoming increasingly significant as a cause of bronchopneumonia and lung abscessation in foals. The organism can survive within macrophages and may thus escape normal pulmonary defence mechanisms, particularly in immunocompromised animals. The disease has hitherto been associated with mortality rates as high as 80 per cent, partly as a result of inappropriate therapy. The selection of lipid-soluble antibiotics capable of intracellular penetration is critical for the successful treatment of C equi lung abscesses. A combination of two such antibiotics, erythromycin (25 mg/kg three times daily) and rifampicin (5 mg/kg twice daily) has been used on foals since 1981. Most of these animals had radiographic evidence of extensive lung abscessation, and in all cases the presence of C equi was confirmed on culture of tracheal aspirates. The duration of therapy ranged from four to nine weeks. Mild gastritis and diarrhoea were occasionally noted, but never such as to require termination of the therapy. No other adverse side effects were encountered. The success rate, as judged by a return to normal of chest radiographs and plasma fibrinogen concentrations, has exceeded 80 per cent.

Actinomycetales Infections↗
PubMed1993

Rhodococcus equi infection in foals: a report of an outbreak on a thoroughbred stud in Zimbabwe.

Twenty-four foals were confirmed to be infected with Rhodococcus equi on a private stud in Zimbabwe over a two-year period. Six mares had foals which were affected in each of the two years. All the foals were febrile and early cases were detected by this pyrexia. Bronchopneumonia was only clinically detectable in advanced cases. In spite of energetic hygiene measures relating to pasture and housing management, the incidence was higher in the second year (23 per cent of foals born) than in the first (15 per cent of foals born). The mean age of the foals was significantly greater in the second year. The immunological status of some of the foals was obtained from zinc sulphate turbidity tests performed at 24 hours old and all the affected foals so tested were considered to have had effective colostral transfer. Clinically, affected foals showed pyrexia, tachypnoea and tachycardia and many had a scanty purulent nasal discharge. Only one animal had diarrhoea which was attributed to enteric infection with R equi. Diagnosis was confirmed by clinical examination, culture of the organism from tracheal aspirates and thoracic radiography and was supported by significant elevations of plasma fibrinogen and platelet and neutrophil counts in all cases. Anaemia was not a major finding in any case. Rifampicin and erythromycin were administered per os and weight-related doses were continued until plasma fibrinogen and the absolute neutrophil and platelet counts had been persistently normal for two weeks. Supportive radiography was obtained in some cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinomycetales Infections↗
PubMed1988

Metabolism of acetylene and acetaldehyde by Rhodococcus rhodochrous.

We studied the ability of a soil bacterium, identified as Rhodococcus rhodochrous, to grow on acetylene and to accumulate acetaldehyde. Its maximum growth rate on acetylene was obtained at about 30 degrees C (mu = 0.11 h-1) and was independent of the concentration of this gas in air from 0.14 to 16% (v/v). During growth, acetylene was quantitatively transformed to acetaldehyde, ethanol, acetate, CO2, and biomass in proportions which varied with culture age and temperature. Growth was completely inhibited by acetaldehyde at a concentration of 10 mM. The inhibitory effect was relieved by addition of acetate. Growth on ethanol up to 140 mM did not result in acetaldehyde accumulation. Acetylene consumption was constitutive with apparent Km and Vmax equal to 250 microM and 800 nmol.min-1.(mg protein)-1, respectively. In resting cell suspensions, acetylene consumption rates decreased more rapidly under air than under nitrogen. The inhibitory effect of acetaldehyde was enhanced in the presence of oxygen. Acetaldehyde accumulation in aerobic resting cell conditions did not exceed 10 mM (440 mg/L), but under anaerobic conditions it attained more than 70 mM (3.08 g/L).

Acetaldehyde↗
PubMed1997

Biodegradation of groundwater pollutants by a combined culture of Mycobacterium vaccae and a Rhodococcus sp.

The catabolism of selected groundwater pollutants by a combined culture of Mycobacterium vaccae and a Rhodococcus sp. (strain R-22) was investigated. The M. vaccae-R-22 combined culture was five times more effective in mineralizing benzene than either organism alone. Mycobacterium vaccae oxidized benzene to phenol, and R-22 catabolized the phenol to cellular components and CO2. Benzene did not support growth of M. vaccae, R-22, or the combined culture. Optimization of ratios of the two species indicated that the maximum mineralization of benzene occurred at an initial ratio of 75% M. vaccae to 25% R-22. Cell fractionation of the combined culture after mineralization of [U-14C]benzene indicated that 10% of the benzene carbon was incorporated into cell material, and of this 45% was present in protein and 20% in nucleic acids. This suggested that minimally one species could utilize the products of benzene as a nutrient source. The M. vaccae-R-22 combined culture catabolized ethylbenzene and chlorobenzene without the accumulation of phenolic intermediates, which are inhibitory to M. vaccae's ability to degrade the parent compounds. This study demonstrates that defined mixed cultures may be useful in studying the effects of environmental pollutant degradation on microbial ecosystems and mineralization of these pollutants by the ecosystem.

Benzene↗
PubMed1990

Structural analysis of the specific capsular polysaccharide of Rhodococcus equi serotype 1.

The specific capsular polysaccharide produced by Rhodococcus equi serotype 1 was found to be a high molecular weight acidic polymer composed of D-glucose, D-mannose, and D-glucuronic acid. Structural analysis of the polysaccharide employed a combination of chemical and nuclear magnetic resonance techniques, from which it was determined that the polysaccharide possessed a linear repeating tetrasaccharide unit containing a single O-acetyl substituent and and acetal-linked pyruvic acid moiety: [formula: see text] The 1H and 13C nuclear magnetic resonances of O-deacetylated and pyruvic-free serotype 1 polysaccharides were fully assigned by homo- and hetero-nuclear chemical shift correlation methods.

Acetylation↗
PubMed2001

Leafy gall formation by Rhodococcus fascians.

Rhodococcus fascians infects a wide range of plants, initiating the formation of leafy galls that consist of centers of shoot amplification and shoot growth inhibition. R. fascians is an epiphyte but it also can establish endophytic populations. Bacterial signals involved in symptom development initiate de novo cell division and shoot meristem formation in differentiated tissues. The R. fascians signals exert activities that are distinct from mere cytokinin effects, and the evidence points to a process that adopted cytokinin biosynthetic enzymes to form derivatives with unique activity. Genes implicated in leafy gall formation are located on a linear plasmid and are subject to a highly controlling, complex regulatory network, integrating autoregulatory compounds and environmental signals. Leafy galls are considered as centers with specific metabolic features, a niche where populations of R. fascians experience a selective advantage. Such "metabolic habitat modification" might be universal for gall-inducing bacteria.

Magnoliopsida↗
PubMed1998

Rhodococcus equi lung abscess complicating Evan's syndrome treated with corticosteroid.

We report a case of Rhodococcus equi, an unusual pathogen, causing a right upper lobe lung abscess in a patient with Evan's syndrome (auto-immune haemolytic anaemia and thrombocytopenia) who was treated with high-dose corticosteroid therapy. The patient was treated successfully with clarithromycin, vancomycin, ciprofloxacin and imipenen which appear to be effective in combination for this unusual condition in which the treatment regimen has been controversial.

Actinomycetales Infections↗
PubMedDate not supplied

Pulmonary malacoplakia associated with Rhodococcus equi infection in an AIDS patient. Report of a case with diagnosis by fine needle aspiration.

BACKGROUND: Malacoplakia is an uncommon, peculiar inflammatory disease with few cytologic descriptions. It is seen mostly in the lower urinary tract, with rare occurrences in the lungs. Escherichia coli is the most common underlying etiologic agent, with only a few cases reported with Rhodococcus equi infection. CASE: A 36-year-old female with AIDS presented with dysphagia, dry cough and fever. Physical examination revealed diffuse bronchi and wheezing. Chest roentgenography showed multiple cavitary lesions bilaterally in the lungs. Fine needle aspiration (FNA) of one of these lesions revealed clusters of granular histiocytes with numerous intracytoplasmic and extracytoplasmic Michaelis-Gutman bodies. Also noted were numerous coccobacillary bacteria, which, on culture, were identified as R equi. Ultrastructural findings are also presented. CONCLUSION: The case illustrates the cytologic findings of malacoplakia in a rare pulmonary lesion associated with R equi infection in an AIDS patient. It reiterates the diagnostic role of FNA in dealing with a nonneoplastic entity for timely and definitive treatment and follow-up.

AIDS-Related Opportunistic Infections↗
PubMed1996

Immunohistochemical detection of virulence-associated antigens of Rhodococcus equi in pulmonary lesions of foals.

Rhodococcus equi was isolated from the lungs of six foals with bronchopneumonia. All isolates expressed 15-17-kd antigens by immunoblot analysis and contained a virulence-associated plasmid of 85 or 90 kb. Immunohistochemically, R. equi from all pulmonary lesions showed the expression of 15-17-kd antigens mainly in the phagocytic cells. The specific monoclonal antibody to 15-17-kd antigens of R. equi (MAb 10G5) may be an aid in the diagnosis of R. equi-induced pneumonia.

Actinomycetales Infections↗