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Rhodococcus equi (R. equi) primarily causes zoonotic infections affecting grazing animals and is an unusual cause of infection in immunocompetent human beings. We report a case of bacteremia due to R. equi a rare isolate in a child suffering from protein energy malnutrition
The in vitro susceptibility of nine Rhodococcus equi strains (seven isolates from immunocompromised patients mainly HIV positive and two reference strains) to twenty various antibiotics were assessed for bacteriostatic effects by an agar dilution method. Imipenem and ceftriaxone were the most effective of the beta-lactams studied. The lowest MIC were noted with vancomycin, teicoplanin, erythromycin, clarithromycin, rifampicin, gentamicin and doxycycline. A longitudinal survey, including three strains isolated from the same patient, showed the emergence of rifampicin resistance and a marked increase of the MIC to imipenem.
Rhodococcus equi, a recognized pathogen in horses, is emerging as a human opportunistic pathogen, especially in immunocompromized hosts. We describe four immunocompromized patients who had serious R. equi infections with an overall mortality of 75%. The natural habitat of R. equi is soil, particularly soil contaminated with animal manure. Necrotizing pneumonia is the commonest form of infection but extrapulmonary infections, such as wound infections and subcutaneous abscess, have also been described in humans. R. equi is cultured easily in ordinary non-selective media. Large, smooth, irregular colonies appear within 48 hours. It is a facultative, intracellular, nonmotile, non-spore forming, gram-positive coccobacillus, which is weakly acid-fast staining and bears a similarity to diphtheroids. It forms a salmon-colored pigment usually after 48 hours incubation. A particular characteristic of this organism is that it undergoes synergistic hemolysis with some bacteria on sheep blood agar. R. equi may be misidentified as diphtheroids, Mycobacterium species, or Nocardia. In vitro R. equi is usually susceptible to erythromycin, ciprofloxacin, vancomycin, aminoglycosides, rifampin, imipenem and meropenem. The organism can be difficult to eradicate, making treatment challenging. Increased awareness of the infection may help with early diagnosis and timely treatment.
The production of biosurfactant by Rhodococcus erythropolis during the growth on glycerol was investigated. The process was carried out at 28 degrees C in a 1.5-L bioreactor using glycerol as carbon source. The bioprocess was monitored through measurements of biosurfactant concentration and glycerol consumption. After 51 h of cultivation, 1.7 g/L of biosurfactant, surface, and interfacial tensions values (with n-hexadecane) of 43 and 15 mN/m, respectively, 67% of Emulsifying Index (E24), and 94% of oil removal were obtained. The use of glycerol rather than what happens with hydrophobic carbon source allowed the release of the biosurfactant, originally associated to the cell wall.
Rhodococcus sp. PR-1 with high capability of demulsification on a surfactant-stabilized kerosene-water model emulsion was isolated from sewage of Dagang oil field. It could demulsify the model emulsion completely in 8 hours at 55 degrees C and had better demulsifying capability than chemical demulsifier DGF-01. The freezing-thawing and autoclaving had no effect on the process of demulsification, yet ultrasonic disposal and deal with organic solvent could inhibit its activity. A linear relationship, not the first order reaction that was used in some references, was observed between the percentage of demulsification and reaction time. The demulsifying capability of PR-1 was mainly resulted from the hydrophobic surfaces of microbial cells, which were characterized by the mycolic acids with the carbon-chain-length from 27 to 54. These results provide foundation for biological application on demulsifying crude oil emulsions in produced water of oil field.
Before highly active antiretroviral therapy (HAART) has become available, antibiotic treatment was usually unable to eradicate Rhodococcus (R.) equi infection in HIV-infected patients, although some clinical improvement could be observed in most cases. There are limited data on the outcome of treatment of R. equi pneumonia in the HAART era. We report on a 52-year-old HIV-infected man who presented in poor general condition with an extensive lung cavitation lesion caused by R. equi. The patient recalled exposure to horses on several occasions. R. equi was cultured from the sputum and the isolate was sensitive to imipenem vancomycin, co-trimoxazole, erythromycin, azithromycin, ciprofloxacin and rifampicin. The CD4+ lymphocyte count was 5 cells/mm3 (0.9%) and his plasma HIV-1 RNA viral load was 101000 copies/mL. The patient was successfully treated with a combination of antibiotics that included azithromycin both as part of an initial and suppressive regimen together with antiretroviral treatment. Surgery was not needed and the patient had no relapse for more than five years after the diagnosis and for more than 3 years of suppressive therapy discontinuation. Our literature search revealed 27 patients treated for R. equi infection in the HAART era. However, details on antimicrobial treatment were given in only 3 cases. The optimal drug regimen and duration of treatment for R. equi pneumonia have not yet been established. Because drug resistance may occur during single agent therapy, it has been suggested that at least two antibiotics to which R. equi is susceptible be given. The recommended choices usually include imipenem, antipseudomonal aminoglycosides, erythromycin or azithromycin, vancomycin, rifampin, and levofloxacin. To our knowledge this is the first documented case of long term remission of R. equi pneumonia in an HIV-infected man treated with azithromycin as part of his antibiotic regimen and HAART.
Infection with Rhodococcus equi is an important cause of pneumonia in foals, but other organ systems may also be affected. The intracellular presence of R. equi and the formation of granulomatous and suppurative inflammatory tissue mean that prolonged treatment is needed. The pharmacological properties of the combination of erythromycin and rifampicin have improved the survival of foals infected with R. equi; however, erythromycin can cause adverse reactions in foals and mares, which has prompted the search for alternative therapies. The combination of azithromycin or clarithromycin with rifampicin seems to be a promising alternative. However these combinations are expensive and adverse effects remain to be determined, especially in the dams of treated foals. Thus correct diagnosis and appropriate use of drugs are essential for the treatment of R. equi infection in foals.
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.
The ability of Rhodococcus rhodochrous strain 172 to consume fluorene as the sole source of carbon and energy in a liquid medium was successfully increased by an addition of polycapramide fiber to the medium and preliminary adaptation of cells on fluorene agar. A combination of these approaches allowed complete degradation of fluorene without an accumulation of intermediates.
The amidase gene from Rhodococcus rhodochrous M18 was cloned by PCR amplification with primers developed by use of peptide amino acid sequences obtained after treating amidase with trypsin. Nucleotide sequence analysis of this gene revealed high homology with aliphatic amidases from R. erythropolis R312 and Pseudomonas aeruginosa. Considering the substrate specificity and the results of DNA analysis, amidase from R. rhodochrous M8 was assigned to the group of aliphatic amidases preferentially hydrolyzing short-chain aliphatic amides. The amidase gene was expressed in cells of Escherichia coli from the self promoter and from the lac promoter. To clone a fragment of R. rhodochrous M8 chromosome (approximately 9 kb), containing the entire structural gene and its flanking regions, plasmid pRY1 that can be integrated into the chromosome via homology regions was used. No sequences of the nitrile hydratase gene, the second key gene of nitrile degradation in strain R. rhodochrous M8, were detected. Thus, genes encoding amidase and nitrile hydratase in strain R. rhodochrous M8 are not organized into a single operon despite their common regulation.
Rhodococcus equi can cause severe or fatal pneumonia in foals as well as in immunocompromised animals and humans. Its ability to persist in macrophages is fundamental to how it causes disease, but the basis of this is poorly understood. To examine further the general application of a recently developed system of targeted gene mutation and to assess the importance of different genes in resistance to innate immune defenses, we disrupted the genes encoding high-temperature requirement A (htrA), nitrate reductase (narG), peptidase D (pepD), phosphoribosylaminoimidazole-succinocarboxamide synthase (purC), and superoxide dismutase (sodC) in strain 103 of R. equi using a double-crossover homologous recombination approach. Virulence testing by clearance after intravenous injection in mice showed that the htrA and narG mutants were fully attenuated, the purC and sodC mutants were unchanged, and the pepD mutant was slightly attenuated. Complementation with the pREM shuttle plasmid restored the virulence of the htrA and pepD mutants but not that of the narG mutant. A single-crossover mutation approach was simpler and faster than the double-crossover homologous recombination technique and was used to obtain mutations in 6 other genes potentially involved in virulence (clpB, fadD8, fbpB, glnA1, regX3, and sigF). These mutants were not attenuated in the mouse clearance assay. We were not able to obtain mutants for genesfurA, galE, and sigE using the single-crossover mutation approach. In summary, the targeted-mutation system had general applicability but was not always completely successful, perhaps because some genes are essential under the growth conditions used or because the success of mutation depends on the target genes.
This study evaluated the prophylactic effectiveness of hyperimmune plasma (HIP) as an aid in the prevention of pneumonia caused by experimental infection with Rhodococcus equi. Thirty neonatal foals were administered R. equi HIP or saline at 2 days of age and were infected with virulent R. equi at 7 days. All foals developed signs or symptoms of respiratory disease. Radiographic scores on day 28 and neutrophil concentrations on day 49 were significantly greater in control foals, and time to respiratory effort score of 2 or higher was significantly shorter for control foals. Three foals, all in the principal group, died or were euthanized before the end of the study, but there was no significant difference in mortality between groups. VapA titers were significantly greater in principal foals. Administration of R. equi HIP decreased the severity of radiographic lesions and prolonged time to increased respiratory effort due to R. equi-induced pneumonia.
Cholesterol oxidase (CO) and choline phosphohydrolase (CPH) exoenzymes were isolated from culture supernatants of Rhodococcus equi ATCC 33701 and their hemolytic and cytotoxic activities examined. The purifications involved differential ammonium sulphate precipitation, ion exchange and gel filtration chromatography. A purification of 32.8-fold and a yield of 0.3% of CO were determined by synergistic hemolysis of sheep red blood cells (SRBC) presensitized with Staphylococcus aureus beta toxin. The enzymatic activity of CO was also demonstrated by oxidation of aqueous cholesterol suspensions. The activity of CO was reversibly inhibited by concentration. A purification of 412.4-fold and a yield of 1.7% of CPH were determined by hydrolysis of p-nitrophenyphosphorylcholine. Purity of both exoenzymes was confirmed by immunoblotting. On sodium dodecyl sulphate polyacrylamide gel electrophoresis, the CO had a molecular mass (Mr) of 60 kd and the CPH a Mr of 65 kd. Choline phosphohydrolase did not hydrolyse sphingomyelin. Sphingomyelinase C (SMC) activity was however demonstrated in concentrated culture supernatants. This dissociation of SMC from CPH activity indicates that R. equi produces two distinct phospholipase C exoenzymes, a CPH and a SMC. Both CO and CPH combined, or individually, did not lyse native SRBC even with subsequent chilling of the cells at 4 degrees C ("hot-cold" treatment). Purified CO lysed beta toxin-sensitized SRBC. The CPH showed only minor hemolytic activity against such sensitized SRBC even at high concentrations. Combination of CO and CPH in lysis of beta toxin sensitized SRBC showed only minor additive rather than synergistic effects.(ABSTRACT TRUNCATED AT 250 WORDS)
A 2-year field study was performed to determine the capability of increasing Rhodococcus equi specific antibody in foals via plasma transfusion or mare vaccination, to determine the kinetics of R. equi (ELISA) antibody decay and to assess the protective effects of these procedures in foals on a farm endemic for R. equi. Plasma donors were vaccinated with a killed R. equi bacterin and produced high levels of anti-R. equi antibodies, which were harvested by plasmapheresis. In Experiment 1, 68 foals were given 1 litre of hyperimmune plasma intravenously (i.v.) between 1-60 days of age. Foal plasma R. equi antibody was significantly increased and high levels of R. equi antibody (ELISA) were maintained for 60 days. No R. equi pneumonia developed in any foals receiving plasma. In Experiment 2, 99 pregnant mares were vaccinated with R. equi bacterin at 30, 60 and 90 days before foaling. Group 1 foals (101:85 from R. equi immunized mares) also received plasma transfusions and Group 2 foals (14), from R. equi immunized mares, did not receive plasma transfusions. Pregnant mare immunization increased colostrum R. equi antibody significantly. Eight foals showed failure of transfer of specific R. equi antibody. The incidence of R. equi pneumonia was 2.9% in Group 1 foals and 43% in Group 2 foals. Vaccination of pregnant mares did not provide protection against R. equi pneumonia; however, plasma transfusion with hyperimmune plasma administered prior to R. equi exposure was significantly protective in foals.
Rhodococcus equi (Corynebacterium equi) is able to produce infections not only in animals but also in patients, usually immunosuppressed ones. We report a new case of bacteremic pneumonia in a drug addict who was also infected by HIV. X-ray film of the chest showed a cavitated infiltrate in right upper lobe. R. equi was recovered from blood, respiratory secretions and lung tissue. The patient was with a prolonged course of antibiotics and also surgical treatment. In spite of both therapies, the patient died. We believe that this infection has to be considered in HIV infected patients with cavitated pneumonia and that early surgical treatment, combined with a prolonged course of multiple antibiotics, is advisable. Finally, in view of the severity of this infection, and its relation with other opportunistic infections, we believe that could be included as AIDS diagnostic criteria.
Three Rhodococcus rubropertinctus dissociants (R, S and M) and two Streptococcus lactis dissociants (R and S) were compared using the electron microscopy method of negative contrasting. The cell wall of R.rubropertinctus dissociants had a thickness of 40 nm (R), 30 nm (S) and 20 nm (M). The cell wall of S. lactis dissociants was 35-55 nm (R) and 25-30 nm (S) thick. 1.5-2-fold variations in the thickness of cell walls could account for the different resistance of the dissociants against the action of such external factors as dehydration, antibiotics, UV, elevated NaCl concentrations, etc.
A male patient of 33 years of age with AIDS developed pneumonia with abscess formation and a slow clinical course. The sputum, bronchoalveolar lavage and blood cultures yielded a Gram-positive, partially acid-alcohol resistant bacillus which was identified as Rhodococcus equi. This organism has been uncommonly implicated in pneumonia in patients with immunodeficiency, particularly with neoplasia and severe cell immunity deficiency. It has been very rarely reported in patients with AIDS. To our knowledge, this is the first reported case in Spain.
A foal with vegetative bacterial endocarditis affecting the wall of the left atrium was treated successfully with cefotaxime, erythromycin, and rifampin. Bacterial isolates included Escherichia coli from blood and Rhodococcus equi from a P-type osteomyelitic lesion of the left third metatarsal bone and from synovial fluid from the left metatarsophalangeal joint. Cardiac complications included cardiomegaly and atrial fibrillation, which responded to treatment with digoxin and quinidine sulfate. Cardiac function was considered normal 18 months after treatment. Bacteriologic cure of osteoarthritis was achieved by use of surgical debridement, lavage, and local and systemic antimicrobial treatment; however, lameness developed 18 months after treatment when training for flat racing was begun. Radiography revealed chronic degenerative joint disease.