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Surgical management of Rhodococcus equi metaphysitis in a foal.

A chronic Rhodococcus equi metaphysitis involving the distal growth plate of the left third metatarsal bone had induced a longstanding lameness in a young foal. Abnormal hematologic values included mild anemia, hyperfibrinogemia, mild leukocytosis, and neutrophilia. Radiography of the distal portion of MT3 revealed a radiolucent zone on the medial aspect of the growth plate, and small pieces of bone suggestive of sequestra. Treatment with erythromycin estolate and rifampin, aggressive surgical debridement, and cancellous bone grafting helped resolve the bone infection.

Abscess↗

[Rhodococcus equi infection in a goat].

Enterotoxaemia caused by Clostridium perfringens was diagnosed in a goat affected with severe enteritis which was found to be present at autopsy. Lesions bearing a resemblance to tuberculosis, caused by a Rhodococcus equi infection, were observed in the liver and lungs. This bacterium is constantly isolated from pulmonary abscesses in foals affected with pyemia. Similar cases of infection in goats were previously reported in the literature.

Abscess↗

Influence of Rhodococcus equi on the respiratory burst of resident alveolar macrophages from adult horses.

Opsonized Rhodococcus equi activated the respiratory burst of resident alveolar macrophages (AM) from adult horses in a logarithmic-linear, mass-related manner. The effect of R equi was not significantly different from that of equal masses of opsonized zymosan A. Therefore, R equi does not appear to attenuate the respiratory burst of equine AM. The stimulatory effect of R equi was not reflected by increased production of superoxide anion (O2-), but increased activity of the hexose monophosphate shunt was observed. These results suggest a similarity between the respiratory burst of AM from horses and that of AM from rabbits. We concluded that resident AM from adult horses do not produce O2- concurrently with an increase in activity of the hexose monophosphate shunt when stimulated with either opsonized zymosan A or opsonized R equi. This suggests that O2- is not an important component of the antibacterial defenses of equine AM. Whether equine AM are incapable of producing O2- or require different stimuli to produce it was not determined.

Animals↗

Purification and characterization of glycosphingolipid-specific endoglycosidases (endoglycoceramidases) from a mutant strain of Rhodococcus sp. Evidence for three molecular species of endoglycoceramidase with different specificities.

Two molecular species of endoglycoceramidase (designated as endoglycoceramidases I and II) were purified 32,700 and 43,000 times with overall recoveries of 4.8 and 2.9%, respectively, from a culture fluid of the mutant strain M-750 of Rhodococcus sp., cultivated in the absence of inducers (ganglioside). After being stained with Coomassie Brilliant Blue or a silver-staining solution, each purified enzyme showed a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. The apparent molecular weights, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were 55,900 for endoglycoceramidase I and 58,900 for endoglycoceramidase II, and their pIs were 5.3 and 4.5, respectively. both were capable of hydrolyzing the glucosylceramide linkage of ganglio-type, lacto-type, and globo-type glycosphingolipids to afford intact oligosaccharides and ceramides. Globo-type glycosphingolipids were strongly resistant to hydrolysis by endoglycoceramidase II in comparison with endoglycoceramidase I. Neither could hydrolyze gala-type glycosphingolipids, cerebrosides, sulfatides, glycoglycerolipids, or sphingomyelins. In addition to these two enzymes, the strain M-750 produced a third minor molecular species of endoglycoceramidase designated as endoglycoceramidase III. It was found capable of specifically hydrolyzing the galactosylceramide linkage of gala-type glycosphingolipids that were not hydrolyzable at all by endoglycoceramidases I or II. The molecular weights of the oligosaccharide and ceramide released from asialo GM1, incubated either in normal H2O or H2(18)O with the enzyme, were compared by fast atom bombardment-mass spectrometry. The result clearly indicated that both endoglycoceramidases I and II hydrolyze the glycosidic linkage between the oligosaccharide and ceramide. Thus, a systematic name of the endoglycoceramidase should be glycosyl-N-acyl-sphingosine 1,1-beta-D-glucanohydrolase.

Enzyme Stability↗

[Rhodococcus equi infection in HIV disease].

A case of Rhodococcus equi pneumonia associated with septicemia and metastatic brain abscess in an HIV-infected male is presented. Clinical findings and the diagnostic and therapeutic approach are discussed. Cavitating pulmonary disease was rapidly improved by six-week combined oral and parenteral antibiotic treatment. However, pulmonary relapse and brain abscess were documented after one month. Therapy with ceftriaxone, ciprofloxacin and cotrimoxazole on an outpatient basis again led to clinical and radiological improvement after four weeks. Surgical resection of localized processes should be considered early, and prolonged antibiotic therapy over months is recommended.

Acquired Immunodeficiency Syndrome↗

Rhodococcus (Corynebacterium) equi: bactericidal capacity of neutrophils from neonatal and adult horses.

The capacity of hematogenous polymorphonuclear neutrophilic leukocytes (PMNL) to kill Rhodococcus equi was compared in horses of various ages. A radioisotope bactericidal assay was used to determine the capacity of PMNL to kill R equi. Assays were conducted on PMNL from horses in 3 groups: group I, 13 foals with a mean age of 3.3 days; group II, 10 group-I foals at a mean age of 35.7 days; and group III, adult dams of group-I foals. Bacteria were obtained from the lungs of a foal with R equi pneumonia and opsonized with fresh adult equine serum that contained R equi specific antibody. The mean peak percentage of R equi killed by PMNL was 78.9 for group I, 90.1 for group II, and 87.9 for group III. There was no significant difference (P greater than 0.05) among groups; however, 15% of foals in group I (2 foals) had a mean peak percentage of 30.5 killed, which was significantly (P less than 0.05) lower than the percentage for other foals in group I. The results of our investigation indicated that the capacity of PMNL to kill opsonized R equi is similar in neonatal, young, and adult horses. However, some neonatal foals have a substantially lower capacity to kill R equi, which may be an important factor in the pathogenesis of R equi infections.

Actinomycetales Infections↗

Protection of foals against experimental Rhodococcus equi pneumonia by oral immunization.

Two groups of three one to three week old foals were immunized orally on four occasions over five weeks with two strains of Rhodococcus equi, a clinical isolate from a pneumonic foal and a laboratory passaged Congo red negative variant of this strain. Three nonimmunized foals of similar age acted as controls. Three weeks after the last immunization, all foals were challenged on five occasions over seven days by aerosol infection with about 10(10) of the pneumonic foal isolate on each occasion. Control foals became seriously ill and were euthanized. Immunization with either strain protected foals equally against the challenge, and resulted in rapid lung clearance. Oral immunization can thus protect foals against severe challenge with R. equi. The proteins associated with Congo red colony staining appear not to be involved in protective immunity.

Actinomycetales Infections↗

Cellulitis and subcutaneous abscesses caused by Rhodococcus equi infection in a foal.

Cellulitis and subcutaneous abscess formation was diagnosed in a 3-month-old Thoroughbred filly. Clinical signs consisted of a large ulcerated plaque, with satellite pustules on the medial aspect of the right hock and subcutaneous abscesses in the right inguinal and mammary gland areas. Laboratory analysis revealed mature neutrophilia. Rhodococcus equi was isolated from the cellulitis and the subcutaneous abscess. Oral administration of erythromycin and rifampin for 35 days resulted in a clinical cure.

Abscess↗

Polyacrylamide gel electrophoresis of whole-cell preparations of Rhodococcus equi.

The whole-cell proteins of ten strains of Rhodococcus equi isolated from horses, pigs, or humans, including the type strain ATCC 6939, were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The protein profiles of seven different capsular serotypes and the type strain were very similar when bacteria were cultured under the same conditions. Protein profiles were largely unaffected by incubation at two temperatures (30 degrees C, 37 degrees C) or times (12 h, 48 h). There were generally minor differences in protein profiles between strains grown in different media (brain heart infusion, nutrient, minca broths, tryptic soy-blood agar) with the marked exception of a prominent diffuse 17.5 kd protein which was expressed in nutrient broth. This protein was not produced by the type strain and was lost on repeated passage in vitro (50th, 100th passage) in two of three other strains examined.

Animals↗

Antibody response of horses to Rhodococcus equi antigens.

The antigens extracted from strains belonging to seven capsular serotypes of Rhodococcus equi, as well as from two wild strains isolated from pneumonic foals, were examined. Whole-cell antigens and soluble products present in broth culture supernatants were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroblotted onto nitrocellulose, and stained with serum from hyperimmunized rabbits or foals. Foal sera used included sera from pneumonic animals with known titer to equi factors; from animals bled monthly on a farm with enzootic pneumonia, and from animals bled monthly on a farm with no history of R. equi pneumonia. The humoral response of foals to somatic antigen preparations was negligible, with few differences noted between sera from healthy, subclinically affected, and sick foals. The humoral response to R. equi broth culture supernatant products appeared more marked and was related to equi factor antibody titer. These findings suggest that the humoral response to R. equi whole-cell antigens is unimportant in protection against disease, which is consistent with the behavior of the organism as a facultative intracellular pathogen.

Animals↗

[Comparison of the sensitivity of Rhodococcus rubropertinctus R, S and M variants to antibiotic action].

Comparison of sensitivity of 63 clones of R, S and M variants of Rhodococcus rubropertinctus 104 to 6 antibiotics showed that sensitivity of the variants was similar only to erythromycin. Their sensitivity to penicillin, streptomycin, chlortetracycline, rifampicin and actinomycin C was different. Still, they preserved similar regularity: M cells were the most sensitive and R cells were the most resistant. There was a 1.5-3 fold difference in sensitivity of R and M variants to the antibiotics. The number and rate of chlortetracycline absorption by R cells were lower than those of S and especially M cells which must condition their higher resistance to the antibiotic.

Anti-Bacterial Agents↗

Quantitative fecal culture for early diagnosis of Corynebacterium (Rhodococcus) equi enteritis in foals.

Quantitative culture of Corynebacterium (Rhodococcus) equi from feces of 17 foals on a farm (A) with an endemic C. equi infection problem and 26 foals on a farm (B) without the disease in the past decade was done with a selective medium at weekly or monthly intervals from April to August of 1984. Corynebacterium equi was observed in the feces of 16 of 17 foals on farm A, and 19 of 26 foals on farm B. The mean viable count of C. equi in one gram of feces was 4.1 +/- 3.7 (log10) on farm A, and 3.9 +/- 3.4 (log10) on farm B. Corynebacterium equi was recovered from feces of foals as young as two weeks old. Almost all foals at an age between two to four weeks shed the bacteria in the feces. During the observation period two foals showed clinical signs: fever, diarrhea, and cough, at four or five weeks old. At the same time the bacterial count per gram of feces increased from 4 to 7 or 8 (log10). They shed large number of bacteria in the feces and continued to show the clinical signs until death at 10 or 11 weeks old. One of the foals was diagnosed as having had C. equi enteritis and pneumonia by the postmortem recognition of lesions with bacteriological confirmation. The quantitative culture of the feces of foals at weekly intervals after birth on farm A was found to be very useful as an aid in early diagnosis of C. equi enteritis in foals.

Actinomycetales Infections↗

[Regulation of terephthalate catabolism in Rhodococcus rubropertinctus].

The regulation of terephthalate catabolism was studied in Rhodococcus rubropertinctus which decomposed this synthetic monomer. The pathway (a) of terephthalate (TP) catabolism is as follows: TP----benzoate----4-hydroxybenzoate----protocatechuate----pyrocatechol-- --cycle ortho-cleavage. The following results were obtained when studying why two other catabolic pathways were realized if benzoate and 4-hydroxybenzoate were taken as a sole carbon source, namely, (b) benzoate----pyrocatechol----cycle cleavage and (c) 4-hydroxybenzoate----protocatechuate----cycle cleavage. TP seemed to cause the divergence of pathways (a) and (b) by repressing the system of benzoate oxidation to pyrocatechol. In pathway (c), benzoate repressed the synthesis of enzymes which catalysed protocatechuate oxidation. Pathway (b) was switched over to (a) when the strain was grown in a medium containing TP and benzoate at a benzoate concentration above 5 mM. Here, the concentration of benzoate (first exogenous and later formed from TP) played a key role. R. rubropertinctus growth in a medium with TP and glucose had diauxic characteristics.

Biodegradation, Environmental↗

Rhodococcus equi pneumonia and pulmonary malakoplakia in acquired immunodeficiency syndrome. Pathologic features.

Pulmonary infection with Rhodococcus equi is rare, and most cases are seen in immunocompromised patients, particularly those with acquired immunodeficiency syndrome. We describe the pathologic features in four cases of culture-positive R equi pneumonia occurring in patients infected with human immunodeficiency virus. All four patients had a solitary cavitary pulmonary mass that was resected (n = 3) or had undergone biopsy (n = 1). Pathologically, all specimens revealed sheets of histiocytes with abundant foamy to eosinophilic cytoplasm with numerous phagolysosomes that were positive for periodic acid-Schiff, Gomori methenamine silver, and Grocott stains. Occasional histiocytes contained Michaelis-Gutmann bodies, diagnostic of malakoplakia. The Michaelis-Gutmann bodies yielded positive results with periodic acid-Schiff, Gomori methenamine silver, Grocott, Giemsa, and von Kossa stains (three of three cases studied) and with alizarin red and Prussian blue stains (two of three cases studied). Many gram-positive coccobacilli within histiocytes and associated with neutrophils were found in one case. Ultrastructural study of one case showed histiocytes containing abundant phagolysosomes with degenerated bacterial components and Michaelis-Gutmann bodies. The latter had a targetoid appearance with variegated phagolysosome cores that were mineralized by deposition of electron-dense spicules surrounded by peripheral rings of granular and membranous material. Based on our observations and reports in the literature, there appears to be a more than coincidental association between pulmonary R equi infection, malakoplakia, and human immunodeficiency virus infection.

Acquired Immunodeficiency Syndrome↗

Infections due to Rhodococcus equi in three HIV-infected patients: microbiological findings and antibiotic susceptibility.

Infections of Rhodococcus equi, a well-known pathogen in animals which causes cavitated pneumonia similar to that caused by mycobacteria, were studied in three HIV-infected patients. This microorganism was isolated in the bronchoalveolar washings of two patients and in the sputum of the third. In two patients, Rh. equi represented the first clinical opportunistic manifestation of HIV disease. One patient died of concomitant Pneumocystis infection. The eradication of the microorganism occurred in two out of three patients. It was found that no isolates were resistant to erythromycin, claritromycin, rifampin, vancomycin, teicoplanin, imipenem, gentamycin or azithromycin (MIC values < or = 0.1 microgram/ml). Moreover, the quinolones (ciprofloxacin and ofloxacin) were found to be less effective, whereas neither the beta-lactam antibiotics nor chloramphenicol were effective therapy for this microrganism. At least two antimicrobial agents should be given contemporaneously to treat these infections for a period of up to several months. Our results suggest that the combinations erythromycin + rifampin or imipenem + teicoplanin are the most effective treatments in Rh. equi infections.

AIDS-Related Opportunistic Infections↗

Molecular characterization of a lipid-modified virulence-associated protein of Rhodococcus equi and its potential in protective immunity.

Virulent strains of Rhodococcus equi produce plasmid-mediated 15- and 17-kDa proteins, which are thermoregulated and apparently surface-expressed. We demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) that R. equi produce three antigenically-related virulence-associated proteins, a diffuse 18-22-kDa, a 17.5-kDa and a 15-kDa protein. Phase partitioning of whole cells of R. equi strain 103 with Triton X-114 (TX-114) and labelling with [3H]-labelled palmitic acid showed that the two higher molecular weight proteins are hydrophobic and lipid modified. The 15-kDa protein did not partition into TX-114 and was not lipid modified. Cloning and expression of a fragment of the R. equi virulence plasmid in Escherichia coli showed that the three proteins were expressed from a single gene. Sequence analysis of this gene (designated vapA) revealed a 570-bp open reading frame encoding a polypeptide of 189 amino acids with a calculated molecular mass of 19,175 Da. The mature, nonlipid modified protein had a calculated mass of 16,246 Da. The 17.5- and 18-22-kDa forms of the protein are therefore due to lipid modification. No significant sequence homology of the vapA gene with other reported nucleotide sequences were found. Opsonization of virulent R. equi with an IgG1 mouse monoclonal antibody (MAb103) to the VapA protein significantly enhanced uptake in the murine macrophage cell line IC-21. Intraperitoneal injection of mice with Mab103 enhanced initial clearance from the liver of mice challenged intravenously with R. equi. Immunization of mice with the lipid-modified VapA purified by SDS-PAGE fractionation or with acetone precipitated VapA protein following TX-114 extraction resulted in significantly enhanced clearance from the liver and spleen following intravenous challenge. The VapA protein of R. equi appears therefore to be a protective immunogen.

Actinomycetales Infections↗

Cauda equina syndrome, diskospondylitis, and a paravertebral abscess caused by Rhodococcus equi in a foal.

A 4-month-old male Quarter Horse was referred for evaluation of urinary incontinence. Physical examination revealed clinical signs consistent with cauda equina syndrome. Radiography revealed diskospondylitis of S2 through S4. Infected bone was surgically curretted, and drainage was established for an associated paravertebral abscess. Rhodococcus equi was isolated from specimens of bone and from fluid samples obtained from the paravertebral abscess. Bethanechol was administered to stimulate urination. Erythromycin and rifampin were administered for 120 days. The foal's neurologic dysfunction resolved completely. Two years after discharge, the horse remained neurologically normal and did not have apparent effects as a result of its previous disorder.

Abscess↗

[Rhodococcus equi pneumonia in patients with immunodeficiency].

In three patients, men of 30, 21 and 24 years old respectively, Rhodococcus equi pneumonia was diagnosed. Antibiotic treatment, followed by secondary prophylaxis, was successful. In one patient, lobectomy was done. R. equi, a well-known bacterial pathogen in horses, appears also to be pathogenic in humans in case of immunodeficiency.

AIDS-Related Opportunistic Infections↗