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Changes in structure of lymphatic organs in foals died due to Rhodococcus equi infection.

The aim of this study was to investigate the structure of lymphatic organs in foals which died due to Rhodococcus equi infection. The material for this study consisted of 9 foals, of both genders, various breeds, 6-14 weeks old. The routine histopathological examination and immunocytochemical reaction for cytokeratine-3 receptors revealed the disturbances in the thymus epithelial cells development. This was manifested by a complete lack of cells expressing cytokeratine-3 receptors, as well as absence of normal thymocytes differentiation. The supposed thymus epithelial cells were very few, expressing faint reaction of cytokeratine-3 receptors. The relative deficiencies in the lymphatic tissue development in peripheral lymphatic organs were observed. It was concluded, that the observed pathological changes may indicate the inherited, not described yet gene failure, which was responsible for the thymus epithelial cells developmental disturbances, and was expressed with various intensity in examined foals. The causes of the fatal course of rhodococcosis in foals were primary disturbances in the immune competence.

Actinomycetales Infections↗

Typing of Rhodococcus equi isolated from submaxillary lymph nodes of pigs in Japan.

Isolation of Rhodococcus equi from the submaxillary lymph nodes of pigs, with or without caseous lymphadenitis, and typing of the isolates by two serological methods were carried out. The rate of isolation of the organisms from the lymph nodes of pigs was 5 times higher in the lymph nodes with caseous lymphadenitis than in those without the lesion. Of 219 isolates, 146 (66.7%) were typable by the method of Prescott, while all the 219 isolates (100%) were typable by the method of Nakazawa et al. The most frequently isolated were serotype 2 of Prescott (identical to serogroup 16 of Nakazawa et al.), and serogroup 3 of Nakazawa et al., which did not correspond with any serotypes of Prescott. Serotypes/serogroups of R. equi from pigs were thus first clarified in Japan.

Actinomycetales Infections↗

A nucleotide mutation associated with fluoroquinolone resistance observed in gyrA of in vitro obtained Rhodococcus equi mutants.

In this study, the quinolone resistance-determining region (QRDR) in gyrA and gyrB of in vitro fluoroquinolone-resistant Rhodococcus equi mutants was sequenced. These mutants were selected from four R. equi strains on blood agar plates containing ciprofloxacin or enrofloxacin. Each mutant became 8- to 64 or greater-fold resistant to fluoroquinolones compared with their parent strains. From the results of sequence analysis of QRDR in gyrA and gyrB, a nucleotide mutation of codon GAC for GGC in gyrA was detected in all mutants, but no mutation was observed in gyrB. This mutation leads to amino acid substitution of Asp for Gly in putative GyrA in R. equi. The position of this substitution corresponds to position 87 of GyrA in Escherichia coli. Our results suggest that the mutation of QRDR in gyrA, which was observed in in vitro fluoroquinolone-resistant R. equi mutants in this study, is closely associated with fluoroquinolone resistance.

Amino Acid Sequence↗

Evaluation of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals.

OBJECTIVE: To determine efficacy of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals. DESIGN: Randomized clinical trial. ANIMALS: 165 foals. PROCEDURE: Foals were randomly assigned to 1 of 2 groups (hyperimmune plasma or nontreated controls). Foals with failure of passive transfer (FPT) of immunity were treated with hyperimmune plasma and evaluated as a third group. Foals that received plasma were given 950 ml between 1 and 10 days of age and between 30 and 50 days of age. A tracheobronchial aspirate was obtained from foals with clinical signs of respiratory tract disease for bacteriologic culture. RESULTS: A significant difference in incidence of pneumonia caused by R equi in foals with adequate passive transfer was not detected between foals that received plasma (19.1%) and nontreated foals (30%). Of 13 foals without FPT that received plasma and developed pneumonia caused by R equi, 12 developed disease prior to administration of the second dose of hyperimmune plasma. Incidence of undifferentiated pneumonia of all causes was not different between groups. CONCLUSION AND CLINICAL RELEVANCE: Intravenous administration of the commercially available hyperimmune plasma was safe, and the product contained high concentrations of anti-R equi antibodies. However, within this limited foal population, the difference in incidence of pneumonia caused by R equi observed between foals that received plasma and control foals was not significant.

Actinomycetales Infections↗

Survival of foals with experimentally induced Rhodococcus equi infection given either hyperimmune plasma containing R. equi antibody or normal equine plasma.

The purpose of this study was to determine if colostrum-deprived foals with experimentally induced Rhodococcus equi pneumonia have a decreased severity of the disease and decreased mortality rate when given hyperimmune (HI) R. equi antibody plasma (R. equi titer at least 100 % and virulence-associated protein A [VapA] at least 10000) prophylactically versus when given normal equine plasma (R. equi titer less than 20 % and VapA less than 160). Sixteen colostrum-deprived foals (R. equi titer less than 5 %) each received normal equine plasma in the first 24 hours of life (R. equi titer less than 20 %). At 14 days of age, six foals were given normal equine plasma and 10 foals were given HI plasma. All foals were subsequently infected intrabronchially with a pathogenic strain of R. equi (2.5 x 10 sup 8; organisms) at 21 days of age. Repeated physical examinations, weight measurements, complete blood cell counts, fibrinogen measurements, and thoracic radiographs (ventrodorsal and lateral) were performed to help determine the severity of the disease. Foals given HI plasma had significantly higher R. equi ELISA titers (42.4 %) than those given normal plasma (20.9 %) on the day of experimental infection. Mortality rates and severity of disease were statistically similar (P >.05) for the groups. Although none of the foals was treated with antibiotics, several with severe R. equi pneumonia recovered. Either HI or normal equine plasma administered to foals in the first few weeks of life caused no adverse effects and may be protective against R. equi, although the exact constituent responsible for protection is undetermined and requires further investigation.

Actinomycetales Infections↗

Rhodococcus equi and Nocardia brasiliensis infection of the brain and liver in a patient with acute nonlymphoblastic leukemia.

Resolution of neutropenia is usually followed by resolution of fever in patients with febrile neutropenia. However, in some cases fever continues even when the patient is no longer neutropenic. Described here is a case of acute myeloblastic leukemia complicated by brain abscess, pulmonary nodules, and hepatic lesions. The patient's fever had continued after the neutropenia resolved; brain and hepatic cultures grew Rhodococcus equi and Nocardia brasiliensis. Although Rhodococcus infections occur frequently in patients with HIV infection, they are uncommon in patients with acute leukemia.

Actinomycetales Infections↗

Prevalence of virulent Rhodococcus equi in isolates from soil and feces of horses from horse-breeding farms with and without endemic infections.

The prevalence of virulent Rhodococcus equi in isolates from soil and feces of foals on a farm with endemic R. equi infections was significantly higher than that of a farm with no history of the disease. Foals bred on a farm with the endemic disease might be constantly exposed to virulent R. equi in their environment.

Actinomycetales Infections↗

Rhodococcus equi infection in HIV-infected patients.

OBJECTIVE: To report clinical and microbiological features and response to treatment in HIV patients with Rhodococcus equi infection. DESIGN: Retrospective study. SETTING: Inpatients admitted to two Infectious Diseases Departments in a community-based hospital. PATIENTS: A total of 12 HIV-positive patients with R. equi infection. MAIN OUTCOME MEASURES: Clinical status, radiological finding, microbiological, haematochemical and immunological tests, and response to treatment. RESULTS: Twelve patients (11 men, six injecting drug users) were diagnosed with R. equi infection. Fever and cough were the principal clinical signs on presentation. Mean CD4+ count at the time of diagnosis was 47.67 x 10(6)/l (SD, 49.2 x 10(6)/l). In 58.3% of the cases the diagnosis of R. equi infection followed the appearance of an AIDS-defining illness. The most frequent radiological findings were cavitary lesions (41.7%) and lung consolidation (33.3%). In 83% of cases, R. equi was isolated from blood and in 33.3% cases from sputum. Test of chemosensitivity showed sensitivity to vancomycin (100%), teicoplanin (100%), ceftriaxone (80%), erythromycin (71%) and ciprofloxacin (66%). Clinical response alone with the disappearance of the presenting signs was observed in nine of the 12 cases (75%); complete response was observed in two cases. Seven patients died with a mortality rate of 58.3% and a mean survival of 5.75 months (SD, 6.48 x 10(6)/l). CONCLUSIONS: R. equi should be considered in the differential diagnosis of pulmonary of disseminated infections in patients with HIV infection. Blood culture may be the most sensitive means of diagnosis. Other studies are needed to determine the most effective choice and duration of antibiotic therapy.

AIDS-Related Opportunistic Infections↗

Expression of virulence-associated antigens of Rhodococcus equi is regulated by temperature and pH.

We recently reported that there are two different virulence-associated antigens correlated with virulence levels in Rhodococcus equi isolates from AIDS patients: virulent R. equi that kills mice with 10(6) cells expresses 15- to 17-kDa antigens and intermediately virulent R. equi that kills mice with 10(7) cells expresses a 20-kDa antigen. Environmental parameters were evaluated for their effects on the expression of these virulence-associated antigens in virulent R. equi strains by immunoblotting using monoclonal antibodies in this study. Expression of these two virulence-associated antigens of R. equi was regulated by pH and temperature; the antigens were produced maximally when the isolates were grown at 38 C and pH 6.5, but were not produced when grown at 38 C and pH 8, nor at temperatures below 30 C. The 20-kDa antigen was found to be located on the cell surface, as were the 15- to 17-kDa antigens, and showed susceptibility to proteolysis by trypsin. These results indicate that expression of the virulence-associated antigens of R. equi is dependent on the environmental conditions.

Antibodies, Bacterial↗

Evaluation of a multiplex polymerase chain reaction assay for simultaneous detection of Rhodococcus equi and the vapA gene.

OBJECTIVE: To evaluate sensitivity and specificity of a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Rhodococcus equi and differentiation of strains that contain the virulence-associated gene (vapA) from strains that do not. SAMPLE POPULATION: 187 isolates of R equi from equine and nonequine tissue and environmental specimens and 27 isolates of bacterial species genetically or morphologically similar to R equi. PROCEDURE: The multiplex PCR assay included 3 gene targets: a universal 311-bp bacterial 16S ribosomal RNA amplicon (positive internal control), a 959-bp R equi-specific target in the cholesterol oxidase gene (choE), and a 564-bp amplicon of the vapA gene. Duplicate multiplex PCR assays for these targets and confirmatory singleplex PCR assays for vapA and choE were performed for each R equi isolate. An additional PCR assay was used to examine isolates for the vapB gene. RESULTS: Results of duplicate multiplex and singleplex PCR assays were correlated in all instances, revealing high specificity and reliability (reproducibility) of the vapA multiplex assay. Of the pulmonary isolates from horses with suspected R equi pneumonia, 97.4% (76/78) yielded positive results for vapA. Seven of 50 (14%) human isolates of R equi yielded positive results for vapA. Six human R equi isolates and 1 porcine isolate yielded positive results for vapB. No isolates with vapA and vapB genes were detected. CONCLUSIONS AND CLINICAL RELEVANCE: The multiplex PCR assay is a sensitive and specific method for simultaneous confirmation of species identity and detection of the vapA gene. The assay appeared to be a useful tool for microbiologic and epidemiologic diagnosis and research.

Actinomycetales Infections↗

Diagnostic contribution of thoracic ultrasonography in 17 foals with Rhodococcus equi pneumonia.

The aim of this retrospective study was to determine the clinical usefulness of thoracic ultrasonography compared to thoracic radiography in evaluation of Rhodococcus equi pneumonia. Criteria for patient inclusion in this study were: (1) isolation of R. equi from transtracheal aspirate, (2) radiographic evaluation of the pulmonary parenchyma, and (3) sonographic evaluation of the pulmonary parenchyma. Seventeen foals met this criteria and their medical records were reviewed. Pyogranulomatous pneumonia was identified radiographically in 13 foals. Severe consolidative pneumonia with no detectable abscessation was identified radiographically in three others. Both consolidation and abscessation were identified radiographically in one. In this foal only consolidation was ultrasonographically identified. Ultrasonographically, pulmonary abscessation was identified in 12 foals and pulmonary consolidation with no detectable abscessation was identified in three others. Sonographic examination allowed detection of only pleural irregularities in one foal, which was subsequently found to have pyogranulomatous pneumonia radiographically. Results indicate that ultrasonography may be an accurate alternative imaging modality for detection of pulmonary pathology attributed to R. equi pneumonia in foals when thoracic radiography is not available.

Actinomycetales Infections↗

Evaluation of a monoclonal antibody-based colony blot test for rapid identification of virulent Rhodococcus equi.

We recently generated a monoclonal antibody immunoglobulin G1 (MAb 10G5), which can recognize 15- to 17-kDa antigens, virulence-associated antigens of Rhodococcus equi, and developed a colony blot enzyme-linked immunosorbent assay with MAb 10G5 for the rapid identification of virulent R. equi. In this epidemiologic study, we evaluated the results of the colony blot test in the identification of virulent isolates of R. equi from feces of horses and soil and compared them with those from a conventional procedure (plasmid profiles of isolates by agarose gel electrophoresis). Environmental isolates (778 isolates from feces of foals, 170 isolates from feces of dams, and 1,267 isolates from soil on horse-breeding farms in Hokkaido) were tested by the colony immunoblot test, and 238 of the 778 isolates, 6 of the 170 isolates, and 85 of the 1,267 isolates showed positive signals. Positive isolates were then analyzed for the presence of virulence plasmid DNA, and 235 (98.7%) of the 238 isolates from foals, 6 (100%) of the 6 isolates from dams, and 75 (88.2%) of the 85 isolates from soil showed the presence of virulence plasmids. On the other hand, 50 isolates from each source, which were randomly selected from the isolates that showed negative signals by colony immunoblot, did not contain virulence plasmids. These results demonstrated that the colony blot test that uses a monoclonal antibody specific for virulence-associated antigens is a rapid and reliable test for the identification of virulent R. equi.

Actinomycetales Infections↗

Diversity of isolates of Rhodococcus equi from Australian thoroughbred horse farms.

Pulsed field gel electrophoresis of restriction endonuclease digested genomic DNA from a collection of clinical isolates of Rhodococcus equi was used to compare strain diversity on different Thoroughbred horse farms over time. Restricted diversity was found among the isolates tested, as the same strains were detected on multiple farms and in multiple years. Marked variation occurred in strain prevalence with some strains being represented by single isolates, and the most prevalent by 26 isolates. There were dominant strains on some farms and the prevalence of some strains differed between farms. Infection with multiple strains was noted in some cases where multiple isolates from a single foal were examined.

Actinomycetales Infections↗

Pathogenicity of Rhodococcus equi strains possessing virulence-associated 15- to 17-kDa and 20-kDa antigens: experimental and natural cases in pigs.

The pathogenic role of Rhodococcus equi in pigs remains controversial. Small numbers of pigs were inoculated intravenously (i.v.), or intramuscularly (i.m.) around the mouth, with a virulent, an intermediately virulent, or an avirulent strain of R. equi and killed 14 days later. None showed clinical signs other than transient fever and weight loss. The virulent and intermediately virulent strains were recovered in culture from various organs and lymph nodes of pigs inoculated i.v., but only from the mandibular lymph nodes of pigs inoculated i.m. The avirulent strain was not recovered from any site. None of the pigs developed macroscopically visible lesions, but they showed reactive hyperplasia of the mandibular lymph nodes. The latter contained scattered phagocytic cells, which were labelled immunohistochemically for virulence-associated antigens (15- to 17-kDa antigens or 20-kDa antigen). Intermediately virulent and virulent strains of R. equi were isolated from mandibular lymph nodes of 5.5% of apparently healthy abattoir pigs (n = 1615). Virulence-associated antigens were detected in phagocytic cells of culture-positive nodes, but the latter showed no lesions other than reactive lymphoid hyperplasia. The results would seem to question the pathogenic role of R. equi in pigs, and it is speculated that the organism survives in the lymph nodes without causing pathognomonic lesions.

Actinomycetales Infections↗

Molecular epidemiology of Rhodococcus equi of intermediate virulence isolated from patients with and without acquired immune deficiency syndrome in Chiang Mai, Thailand.

We investigated the prevalence of virulent Rhodococcus equi in clinical isolates from 69 sporadic cases (60 men, 8 women, and 1 patient of unknown sex) in Chiang Mai, Thailand, between 1993 and 2001. Fifty were human immunodeficiency virus (HIV) positive, 3 were HIV negative, and HIV status was unknown for 16. Fifty-two (75%) of 69 isolates were strains of intermediate virulence that contained the virulence-associated 20-kDa antigen, and 17 isolates (25%) were avirulent. No virulent strains with the virulence-associated 15-17-kDa antigens were identified. R. equi was isolated from HIV-positive patients' houses and those of their neighbors: avirulent strains were widespread, but only 1 strain of intermediate virulence was isolated. R. equi strains of intermediate virulence were isolated from 4 (0.8%) of 500 submaxillary lymph nodes from apparently healthy pigs in Chiang Mai. The routes of R. equi acquisition should be investigated from the viewpoint of zoonosis and public health.

Acquired Immunodeficiency Syndrome↗

Cholesterol oxidase from Rhodococcus equi is likely the major factor involved in the cooperative lytic process (CAMP reaction) with Listeria monocytogenes.

The CAMP reaction between Listeria monocytogenes and Rhodococcus equi was studied by a diffusion assay. Listeria monocytogenes displayed identical cooperative haemolytic effect with supernatant cultures of R. equi or with commercial cholesterol oxidase (COX). This result, even with enzymes of different sources (commercial COX is obtained from Pseudomonas spp.) suggests that this enzyme secreted by R. equi has a crucial role in the synergistic haemolytic (CAMP) reaction with L. monocytogenes. The mechanism of the cooperative lytic process between L. monocytogenes and R. equi may represent a different and novel mechanism reaction, in which the COX may not act as a conventional second-step factor, and a reaction different to the direct interaction with the cholesterol of the erythrocyte membrane may be involved.

Cholesterol Oxidase↗

Rhodococcus equi infection in transplant recipients: a case of mistaken identity and review of the literature.

The incidence of Rhodococcus equi infection in solid-organ transplant recipients continues to rise throughout the world. Unfortunately, this opportunistic pathogen is still underestimated and potentially disregarded by physicians and microbiology laboratories due to its morphology on Gram staining. Pulmonary involvement is the most common finding in the immunocompromised host. We report a case of a 63-year-old heart-transplant recipient who presented with increasing fatigue and nonproductive cough for 3 weeks. After full evaluation, a lung abscess was demonstrated by thoracic computerized tomography (CT). Blood and sputum cultures were remarkable for heavy "diphtheroids." Although the Gram-stain result was initially interpreted as a contaminant, a clinical suspicion for Rhodococcus assisted in further investigation. Broncheoalveolar lavage and CT-guided biopsy of the lung abscess revealed heavy growth of diphtheroids. However, further evaluation by a reference laboratory demonstrated mycolic acid staining consistent with R. equi. Surgical drainage and prolonged antibiotic therapy resulted in complete remission of the pneumonia and abscess. This represents the fourth reported case of R. equi infection in a heart transplant recipient. It is imperative that all physicians and laboratory staff consider R. equi when an immunocompromised patient has any type of pneumonia, especially with abscess formation.

Actinomycetales Infections↗

In vitro activities of polycationic peptides alone and in combination with clinically used antimicrobial agents against Rhodococcus equi.

The in vitro activities of magainin II, nisin, and ranalexin alone and in combination with other antimicrobial agents against six clinical isolates of Rhodococcus equi were investigated by MIC and time-kill studies. All isolates were more susceptible to nisin. A positive interaction was observed when the peptides were combined with ampicillin, ceftriaxone, rifabutin, rifampin, azithromycin, clarithromycin, and vancomycin.

Anti-Bacterial Agents↗