Rhodococcus equi pneumonia in a child with leukemia.
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Quantitative culture of R. equi in the feces of dams and foals, in the air of the stalls and in the soil of the paddocks was carried out on three horse-breeding farms during the foaling season. The isolation rates of R. equi from the feces of dams from the 3 farms suddenly increased to approximately 80% at the end of March, when the snow in the paddocks finished melting, and remained at that level during April and May. The mean number of R. equi and the isolation rate of R. equi from the feces of dams on the farms were investigated for 5 weeks before and 5 weeks after delivery. During the 10 weeks, there were no differences in the isolation rate or in the mean number of R. equi from the feces of dams. R. equi was first isolated from the feces of the foals born in February and the middle of March at 3-4 weeks of age, on the other hand, it was first isolated from the feces of foals born in the end of March and April at 1-2 weeks of age. The number of R. equi in the soil collected from the paddocks used by dams during the winter was approximately 10(2)-10(4) g-1 of soil during the experiment. R. equi was isolated from the air in the stalls at the end of March and the number of R. equi in the air increased particularly on dry and windy days.(ABSTRACT TRUNCATED AT 250 WORDS)
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Normal horses of all ages regularly show evidence of having responded immunologically to R. equi, thus adding serological support to epidemiological evidence that this organism is a normal intestinal inhabitant. More animals from "diseased" farms show a stronger antibody response when compared with foals from "healthy" farms. Various serological tests have been used to detect evidence of infection and to relate antibody level to severity of disease. Anti-R. equi IgG antibody levels, as measured by ELISA, are raised significantly during natural infection. Clinical severity of pneumonia can be correlated with lower specific antibody responses. Following experimental infection, immunological responses can be detected by complement fixation, indirect immunofluorescence, ELISA, lymphocyte blastogenesis and skin testing. Very little work has been carried out to evaluate vaccines against R. equi infection and results have not been encouraging. Success in treatment has been reported following passive immunisation. Administration of immune leucocyte extracts has had no effect on morbidity or mortality rates. The widespread distribution of this organism, together with the relative infrequency of disease caused by it, suggest that R. equi may initiate infection only in such circumstances as a very high infectious challenge, immunological immaturity or deficiency in the host and genetic predisposition.
We studied the inhibitory effect on the PCA reaction of BSA-rabbit anti-BSA (BSA-anti-BSA) system in CP-sensitized and CP-unsensitized guinea pigs by the oral administration of C. equi. Strain Ko-85 (Ko-85) cells. The results were as follows: 1) Guinea pigs were sensitized by intraperitoneal injection with the CP. Nine days later, the animals were administered with an oral dose of 80 mg (W.W.) of Ko-85 cells. The animals were submitted daily to PCA reaction tests using BSA and rabbit anti-BSA serum. The inhibition of PCA reaction of BSA-anti-BSA system in CP-sensitized guinea pigs was shown from 4th day to 10th day after administration of Ko-85 cells. The PCA reaction of 9th day was recognized strong inhibition by the oral administration of Ko-85 cells. 2) CP-unsensitized guinea pigs were given an oral dose of 80 mg of Ko-85 cells, and the animals were submitted every other day to PCA reaction tests of the BSA-anti-BSA system. PCA elicited in these animals on the 9th, 15th and 17th days Ko-85 administration were inhibited compared with those in the control animals (which were not given an oral dose of Ko-85 cells). The strongest inhibition was shown in animals challenged on the 15th day after Ko-85 administration. The inhibitory effect of PCA reaction of BSA-anti-BSA system by the oral administration of an oral dose 80 mg (W.W.) of Ko-85 cells was recognized in CP-sensitized and CP-unsensitized guinea pigs.
A procedure was developed for isolating large numbers of purified polymorphonuclear leukocytes (PMNs) from the peripheral blood of horses. Equine PMN function was evaluated by three procedures: 1) Staphylococcus aureus ingestion, 2) nitroblue tetrazolium reduction, and 3) iodination. Four preparations of R. equi were added to polymorphonuclear leukocytes (PMNs) in each test system. Live bacteria, heat-killed bacteria, the washed pellet from heat-killed bacteria, and the supernatant fluid from heat-killed bacteria were evaluated for effects on equine PMN function. None of the R. equi preparations had an effect on S. aureus ingestion by equine PMNs. Nitroblue tetrazolium reduction by PMNs, a measure of oxidative metabolism, was suppressed by pellet and supernatant fractions. Values for the iodination reaction were depressed by all R. equi preparations, indicating decreased activity of the myeloperoxidase-H2O2-halide system of the PMN. Further evaluation of the supernatant from heat-killed R. equi showed that it retained its inhibitory effect on iodination following autoclaving and/or passage through a 10,000 MW filter. R. equi fractions did not alter the enzymatic conversion of 125I to a protein-bound form in a PMN-free assay developed to evaluate this reaction. The presence of a surface component capable of inhibiting bactericidal mechanisms of the PMN may play an important role in intracellular survival of R. equi.
Two techniques were developed to detect antibodies against the exosubstance of C. equi called equi-factor. In the first technique serum samples are tested against native equi-factor produced by the growth of C. equi on agar medium. A positive result is manifested by the development of precipitation lines. The second test is based on neutralization of prepurified equi-factor by antibody, resulting in the inhibition of its hemolytic synergism with staphylococcal beta toxin. Sera (125 samples) from horses of different ages, kept in localities with a history of C. equi infections, were examined. The first technique detected 65.6%, and the second 40% of positive cases.
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The selective isolation of R. equi in NANC medium was made by quantitative culture of the organism in the feces. R. equi was observed in the feces of all the mares and foals investigated. The mean viable count of R. equi organisms in 1 gram of feces was 8.42 X 10(2) in the mares and 7.57 X 10(2) in the foals, and its 95% confidence limits were in a range of 6.48 X 10(2) to 1.09 X 10(3) and 4.19 X 10(2) to 1.37 X 10(3), respectively. This result indicates that R. equi is a member of the normal intestinal flora in the horse.
A 7-year-old horse with no previous history of illness was determined to have a systemic infection of Rhodococcus equi. Rhodococcus equi was isolated from blood, tracheal fluid, and feces. Lymphopenia, failure to respond to concanavalin A and phytohemagglutinin lymphocyte stimulation testing, decreased concentrations of immunoglobulin (Ig)M, IgA, and IgG, low R equi antibody titer, histologic depletion of lymphoid tissue, and a failure to respond to antigenic stimulation led to the conclusion that both humoral and cell-mediated immunity were compromised. No cause for the acquired immunodeficiency could be determined.
Rhodococcus equi is an intracellular pathogen of macrophages, causing disease in young foals, humans, and sporadically other animals. Although R. equi is easy to grow and manipulate, the analysis of virulence is hampered by a lack of molecular tools. This paper describes the development of a number of versatile plasmids for use in R. equi. Plasmids pREV2 and pREV5 use origins of replication derived from the Mycobacterium fortuitum plasmids pAL5000 and pMF1. These plasmids and their derivatives are compatible in R. equi, allowing their use for analysis of gene function in trans. The stability of these plasmids in R. equi in the absence of selection for the plasmid borne antibiotic resistance markers, and their integrity following passage through Escherichia coli and R. equi was determined.