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[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↗

Immunohistochemical detection of virulence-associated Rhodococcus equi antigens in pulmonary and intestinal lesions in horses.

Two horses with Rhodococcus equi infection were examined post mortem by an immunohistochemical method (peroxidase-antiperoxidase; PAP) with a monoclonal antibody (Mab 10G5) to the 15-17 kDa antigen of R. equi. One of the horses was also examined bacteriologically, R. equi being isolated in culture. Immunolabelling with this Mab was marked and widespread. On the other hand, the immunohistochemical reactivity of infected macrophages with a polyclonal antibody specific for lysozyme was slight. Thus, Mab 10G5 would appear to be a useful diagnostic reagent in R. equi infection, with or without cultural confirmation.

Actinomycetales Infections↗

Associations between the ecology of virulent Rhodococcus equi and the epidemiology of R. equi pneumonia on Australian thoroughbred farms.

The ecology of virulent strains of Rhodococcus equi on horse farms is likely to influence the prevalence and severity of R. equi pneumonia in foals. This study examined the association between the ecology of virulent R. equi and the epidemiology of R. equi pneumonia by collecting air and soil samples over two breeding seasons (28 farm-year combinations) on Thoroughbred breeding farms with different reported prevalences of R. equi pneumonia. Colony blotting and DNA hybridization were used to detect and measure concentrations of virulent R. equi. The prevalence of R. equi pneumonia was associated with the airborne burden of virulent R. equi (both the concentration and the proportion of R. equi bacteria that were virulent) but was not associated with the burden of virulent R. equi in the soil. Univariable screening and multivariable model building were used to evaluate the effect of environmental and management factors on virulent R. equi burdens. Lower soil moisture concentrations and lower pasture heights were significantly associated with elevated airborne concentrations of virulent R. equi, as were the holding pens and lanes, which typically were sandy, dry, and devoid of pasture cover. Few variables appeared to influence concentrations of virulent R. equi in soil. Acidic soil conditions may have contributed to an elevated proportion of virulent strains within the R. equi population. Environmental management strategies that aim to reduce the level of exposure of susceptible foals to airborne virulent R. equi are most likely to reduce the impact of R. equi pneumonia on endemically affected farms.

Actinomycetales Infections↗

[Vertebral osteomyelitis caused by Rhodococcus equi in a three-and-half-month-old Dutch Warmblood foal].

Rhodococcus equi infection occurs worldwide and is especially a problem in foals, where it often causes colitis or pneumonia. Other organs are seldom affected, and their involvement is regarded as a complication of pneumonia and/or colitis. Vertebral osteomyelitis is one such rare complication and is probably caused by haematogenous spread from inflammatory lesions in the lungs and/or intestine. In rare cases, osteomyelitis can be caused by contamination of a wound. This case study describes a foal with vertebral osteomyelitis due to R. equi in which there were only minor inflammatory changes in a mesenteric lymph node.

Actinomycetales Infections↗

Serogrouping of Rhodococcus equi.

The serological relationships among 27 isolates of Rhodococcus equi selected from a total of 1,195 isolates were investigated by cross-agglutination and absorption tests. The presence of capsular material was demonstrated in all the 27 isolates by electron microscopic observation. Antisera were prepared by employing formalized antigen of each isolate. In the cross-agglutination test with formalized antigen, 13 antisera reacted with homologous antigens alone, but the remaining 14 antisera reacted not only with homologous antigens but also with one to four heterologous antigens. When these 14 antisera possessing heterologous agglutinins were absorbed with each of the cross-reacting antigens, 14 specific antisera were obtained. The cross-agglutination test with these 27 antisera proved the 27 strains examined to be serologically distinct from one another. These strains were designated serogroups 1 to 27. Thus the same number of grouping antisera were prepared. The distribution of each serogroup among the 1,195 isolates and 15 reference strains was investigated by the slide agglutination test. All the strains were found to be groupable. Most of them belonged to serogroups 1 to 4, 7 to 9, 11, 14, and 15. Of the serogroups designated, 4, 16, 2, 12, 21, 1, and 9 were identical with Prescott's serovars 1, 2, 3, 4, 5, 6, and 7, respectively.

Agglutination Tests↗

Pathogenicity of Rhodococcus equi expressing a virulence-associated 20 kDa protein (VapB) in foals.

Rhodococcus equi strains of intermediate virulence (IMV) for mice possess a 20kDa protein designated Virulence Associated Protein B (VapB) and a virulence plasmid of 79-100kb, and can be recovered from the submaxillary lymph nodes of pigs. The pathogenicity of such R. equi strains for foals is unknown. In this study, two foals, 42 and 43 days of age, were infected intratracheally with 10(6) and 10(9) cells of R. equi IMV strain A5, respectively. The foal infected with 10(9) cells of strain A5 became clinically ill, with the onset of illness (pyrexia and depression) occurring 21 days after inoculation. R. equi was isolated from the feces and tracheal washings of the foal from 14 to 28 days after inoculation. The foal infected with 10(6) cells of A5 showed no clinical signs, and no R. equi was isolated from any of the samples of feces or tracheal washings during the 28 days of observation. Two foals of 45 and 50 days of age were infected with 10(5) or 10(6) of virulent R. equi ATCC 33701 having 15-17kDa surface proteins designated VapA. Both exhibited severe clinical signs (pyrexia, depression and anorexia) at 12 and 13 days after inoculation. Histopathological examination revealed that strain A5 caused focal granulomatous pneumonia in the foals. R. equi IMV strain A5 was isolated from lung lesions of both foals and from the contents of the intestinal tracts of the foal infected with 10(9) bacteria. These results suggest that IMV R. equi having VapB is less virulent than virulent R. equi having VapA in foals. This finding supports our previous results on the pathogenicities of R. equi strains having these virulence-associated antigens assessed by mouse pathogenicity tests.

Animals↗

Serotaxonomical analyses of strains referred to Nocardia amarae and Rhodococcus equi.

Four strains of Nocardia amarae and four strains of Rhodococcus equi (earlier designated Corynebacterium equi) were analysed by means of the comparative immunodiffusion technique for taxonomical purposes. Nineteen reference precipitation systems, representing different species of Corynebacterium, Mycobacterium, Nocardia, Rhodococcus, Streptomyces, and related taxa were employed. The N. amarae strains differed serologically from the reference strains of Nocardia and their assignment to the latter genus was therefore questioned. Furthermore, the results provided additional evidence for the view that R. equi belongs to the genus Rhodococcus and not to the genus Corynebacterium.

Corynebacterium↗

A physical map of the 85 kb virulence plasmid of Rhodococcus equi 103.

A physical map of the 85 kb virulence plasmid pOTS from Rhodococcus equi 103 was constructed. The restriction map contains 2 AsnI, 5 BglII, 9 EcoRI, 4 HindIII, and 3 XbaI sites. The positions of the EcoRI and HindIII of pOTS are identical to that of the 85 kb virulence plasmid of R. equi ATCC 33701 reported recently by others. EcoRI restriction fragment sizes were similar in the 85 kb plasmids isolated from 4 horse derived R. equi but, except apparently for the 28.3 and possibly 2.0 and 1.5 kb fragments, were different in an 80.1 kb plasmid isolated from a pig source R. equi.

Animals↗

Assessment of the immunogenic potential of Rhodococcus equi virulence associated protein (VapA) in mice.

The development of immunity to Rhodococcus equi, particularly to a virulence-associated protein (VapA) based antigen preparation, was examined in CD1 and BALB/c mice after intraperitoneal vaccination. Immunization with VapA based antigen without adjuvant markedly enhanced organ clearance in CD1 mice but not in BALB/c mice. Delayed type hypersensitivity response and antibody titres in VapA based antigen immunized BALB/c mice were less than in CD1 mice. By contrast also to CD1 mice, sera from immunized BALB/c mice did not react as strongly with VapA in western blots. Use of adjuvants (aluminium hydroxide, iscoms) interfered markedly with the immunogenic properties of the VapA based antigen, in the case of aluminium hydroxide by apparently driving a Th2 type of response. Unexpectedly, iscom adjuvants also impaired immunity and, despite the highest DTH response, produced a low IgG2a response, suggesting that iscomization of the antigen produced a low interferon gamma and high interleukin 2 response. Passive immunization of BALB/c mice with serum from mice immunized with live virulent strain 103+ resulted in only temporary and slight enhancement of organ clearance, supporting the central importance of cellular immunity to R. equi. Immunization with live virulence plasmid- and VapA-positive R. equi strain 103 resulted in marked liver clearance, in marked DTH response and high antibody titres. By contrast, immunization with live virulence plasmid- and VapA-negative strain 103 resulted in slight but variable enhancement of clearance, but insignificant DTH and antibody. The virulence plasmid, and by implication VapA, was thus shown to be critical in determining a highly effective protection to live organisms.

Actinomycetales Infections↗

Epidemiologic study of results of pulsed-field gel electrophoresis of isolates of Rhodococcus equi obtained from horses and horse farms.

OBJECTIVE: To compare isolates of Rhodococcus equi on the basis of geographic source and virulence status by use of pulsed-field gel electrophoresis (PFGE). SAMPLE POPULATION: 290 isolates of R equi (218 virulent isolates from foals and 72 avirulent isolates from feces, soil, and respiratory tract samples) obtained between 1985 and 2000 from horses and horse farms from 4 countries. PROCEDURE: DNA from isolates was digested with the restriction enzyme Asel and tested by use of PFGE. Products were analyzed for similarities in banding patterns by use of dendrograms. A similarity matrix was constructed for isolates, and the matrix was tested for nonrandom distributions of similarity values with respect to groupings of interest. RESULTS: There was little grouping of isolates on the basis of country, virulence status, or region within Texas. Isolates of R equi were generally < 80% similar, as determined by use of PFGE. Isolates from the same farm generally were rarely of the same strain. CONCLUSIONS AND CLINICAL RELEVANCE: Considerable chromosomal variability exists among isolates of R equiobtained from the same farm, sites withinTexas, or among countries from various continents. Only rarely will it be possible to link infections to a given site or region on the basis of analysis of isolates by use of PFGE of chromosomal DNA.

Actinomycetales Infections↗

Structural elucidation of the specific capsular polysaccharide of Rhodococcus equi serotype 7.

Structural analysis of the specific capsular polysaccharide produced by Rhodococcus equi serotype 7 indicated it to be a high-molecular-weight polymer consisting of equal molar amounts of D-galactose, D-mannose, L-rhamnose, and pyruvic acid. By employing a combination of chemical and NMR techniques, it was established that the polysaccharide is composed of the linear repeating trisaccharide units: [formula: see text] -->3)-alpha-D-Galp-(1-->3)-alpha-D-Manp-(1-->3)-alpha-L-Rhap -(1-->, in which the cyclic pyruvic acid acetal groups bridging the O-4 and O-6 positions of the alpha-D-Manp residues have the S-configuration. The 1H and 13C NMR spectra of the native and pyruvic acetal-free polysaccharides were fully assigned.

Bacterial Capsules↗

Facile synthesis of arabinomannose penta- and decasaccharide fragments of the lipoarabinomannan of the equine pathogen, Rhodococcus equi.

Pentasaccharide repeating unit 20 of the lipoarabinomannan from the equine pathogen, Rhodococcus equi, and its dimer 31, were synthesized. The pentasaccharide was obtained by assembling a benzoylated 2,6-branched mannosyl trisaccharide acceptor 13 with a free hydroxyl group at C-2' of the mannose residue attached to the core mannose residue by (1 --> 6)-linkage, followed by coupling with 2,3,5-tri-O-benzoyl-alpha-D-arabinofuranosyl-(1 --> 2)-3,4,6-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroacetimidate (18), and by deacylation. Meanwhile, the decamer 31 was obtained by firstly preparing a benzoylated mannose (1 --> 6)-linked tetrasaccharide backbone 26 with 2-, 2"-O-ClAc, and 2'-, 2'''-O-Ac groups, respectively, then by dechloroacetylation and subsequent condensation with perbenzoylated trichloroacetimidate, and then by deacetylation and subsequent coupling with 18, and finally, by deacylation.

Acetamides↗

Immune response to the Rhodococcus equi infection in high and low antibody-producing mice (Selection IV-A).

Rhodococcus equi is a gram-positive, facultative intracellular bacterium which infects macrophages and causes rhodococcal pneumonia and enteritis in foals. Recently, this agent has been recognized as an opportunistic pathogen for immunocompromised humans. Several murine experimental models have been used to study R. equi infection. High (H(IV-A)) and Low (L(IV-A)) antibody (Ab)-producers mice were obtained by bi-directional genetic selections for their ability to produce antibodies against sheep and human erythrocytes (Selection IV-A). These lines maintain their phenotypes of high and low responders also for other antigens than those of selection (multispecific effect). A higher macrophage activity in L(IV-A) mice has been described for several intracellular infectious agents, which could be responsible for their intense macrophage antigens (Ag)-handling and low Ab production. Due to these differences, L(IV-A) mice were found to exhibit a better performance to trigger an effective immune response towards intracellular pathogens. The objective of this work was to characterize the immune response of Selection IV-A against R. equi. H(IV-A) and L(IV-A) mice were infected with 2.0x10(6) CFU of ATCC 33701+R. equi by intravenous route. With regards to bacterial clearance and survival assays, L(IV-A) mice were more resistant than H(IV-A) mice to virulent R. equi. L(IV-A) mice presented a higher hydrogen peroxide (H2O2) and nitric oxide (NO) endogenous production by splenic macrophages than H(IV-A) mice. L(IV-A) expressed the most intense cellular response, available by the Delayed-Type Hypersensitivity (DTH) reaction, which activated macrophages and produced more H2O2 and NO. The three times higher specific antibodies titres in H(IV-A) indicated that Selection IV-A maintained the multispecific effect and the polygenic control of humoral and cellular responses also to R. equi.

Actinomycetales Infections↗

Blastogenic response of lymphocytes from foals infected with Rhodococcus equi.

The blastogenic response of lymphocytes from 16 newborn foals naturally infected with Rhodococcus equi was investigated, in order to evaluate the relationship between R. equi infection and depressed host response. Naturally infected foals showed evidence of R. equi infection at 5-6 weeks of age, as determined by clinical, haematological, bacteriological and serological methods. The blastogenic response of lymphocytes against phytohaemagglutinin was significantly depressed (stimulation index < 1.80; P < 0.01, P < 0.05) in R. equi-infected foals at 5-6 weeks of age compared with those of control foals. Serum IgG concentration decreased rapidly after foals reached 1 week of age, and minimum levels of IgG were observed at 5-7 weeks of age in R. equi-infected foals. This study suggests that the onset of R. equi infection may be associated with the depressed immune function of naturally infected foals during the first 5-6 weeks after birth.

Actinomycetales Infections↗

The absence of Rhodococcus equi in Mongolian horses.

In native Mongolian horses, the incidence and distribution of Rhodococcus equi are poorly understood. One hundred and fourteen equine fecal samples and 71 soil samples were collected from the camp sites of 26 nomadic families located in three areas less than 100 km from Ulaanbaatar, Mongolia. Five fecal samples were also collected from foals of Przewalski's Horses introduced into the Hustai National Park, Mongolia. No R. equi was isolated from the Mongolian horses or the soil samples. However, three colonies of R. equi were isolated from two fecal samples collected from foals of Przewalski's Horses. These isolates were avirulent, with neither 15- to 17-kDa antigens (VapA) nor a 20-kDa antigen (VapB) genes being detected. We concluded that native Mongolian horses and their environment appear free from contamination with R. equi.

Animals↗

Failure of pulmonary clearance of Rhodococcus equi infection in CD4+ T-lymphocyte-deficient transgenic mice.

Pulmonary clearance of Rhodococcus equi requires functional T lymphocytes. In this study, CD8+ T-lymphocyte-deficient transgenic mice cleared virulent R. equi from the lungs while infection in CD4+ T-lymphocyte-deficient transgenic mice persisted. Although both CD4+ and CD8+ T cells function early in pulmonary defense against R. equi, clearance is dependent on CD4+ T lymphocytes.

Actinomycetales Infections↗

Recurrent Rhodococcus equi infection with fatal outcome in an immunocompetent patient.

The majority of human Rhodococcus equi infections occur in immunocompromised hosts, especially those with AIDS, and infection in immunocompetent patients is rare. Reported here is a case of R. equi infection in a seemingly healthy patient with a very complicated course. Despite neurosurgery and prolonged antibiotic therapy the patient deceased.

Actinomycetales Infections↗