The treatment of pneumonia in foals caused by Rhodococcus (Corynebacterium) equi.
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The pharmacokinetic parameters of a powder formulation of trimethoprim/sulphachlorpyridazine were studied in eight healthy horses which received 5 mg/kg trimethoprim and 25 mg/kg sulphachlorpyridazine 12-hourly with concentrate for five days. The intake of the medicated concentrate by the horses was variable during the first two days, but after they became accustomed to the taste the intake by all the horses during the last three days was good. Faecal samples taken before and on the last day of the drug administrations were negative when cultured for salmonella. Compared with the results of a previous single-dose experiment, higher plasma concentrations and a higher area under the curve for both the drugs were observed. The repeated doses provided plasma concentrations above the minimal inhibitory concentration for Streptococcus zooepidemicus, S equi, Actinobacillus equuli and Rhodococcus equi isolated from the respiratory tract of horses. Synergism between the two drugs occurred at different drug concentration ratios with different bacterial species.
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Undifferentiated distal respiratory tract disease (nasal discharge, cough, pneumonia) in foals (1 to 8 months old) is a burdensome economic problem on breeding farms; yet, the infective agents associated with these episodes have not been well described. Possible causes of these episodes of illness were investigated by culturing specimens of proximal and distal airways of clinically diseased foals (n = 101), prior to any treatment, for aerobic and anaerobic bacteria and viruses (rhinoviruses, equine arteritis virus, equine herpesvirus subtype 1 [EHV-1], influenza virus, and adenovirus). Pairs of sera (n = 47) were examined for antibodies to influenza A virus, equine subtypes 1 and 2, EHV-1, and adenovirus antigens, and sera obtained from foals during acute infection were examined for antibodies (by agar gel immunodiffusion [AGID]) to equi factor antigens of Rhodococcus equi. Viruses were not isolated from the proximal (swab) or distal (bronchial lavage) airway specimens in foals, and only 2 of 47 randomly selected foals seroconverted to EHV-1. Serotiters to the other viruses were low and frequently decreasing between samples, which was compatible with maternally derived antibody. Streptococcus zooepidemicus was the predominant isolate from bronchial lavage specimens (88/101 cases), accompanied by alpha-hemolytic streptococci (8 cases), Bordetella bronchiseptica (13 cases), Staphylococcus epidermidis (9 cases), and other organisms in lesser frequency. Only Str zooepidemicus was recovered significantly (P < 0.05) more often in cases than in controls. The AGID test was found useful to detect foals with presumed exposure to R equi, but positive tests results did not correspond well with bacterial culture results; positive AGID results were recorded in 34% of culture-negative foals.(ABSTRACT TRUNCATED AT 250 WORDS)
In order to assess the Rhodococcus equi infection in three provinces of Turkey (Bursa, Izmir and Istanbul), 696 sera from healthy foals and adult horses were tested by indirect ELISA using a R. equi reference strain (ATCC 6939) as antigen. 103 sera (14.80%) with titres >0.646 resulted positive. Seroprevalence was significantly higher (P=0.0053) in male than in female horses of Istanbul province, although higher antibody titres (mean value) were observed in the female group of Bursa and Izmir provinces with differences estimated between provinces (P=0.0002). Seroprevalence was correlated with age: foals aged less than 1 year (P<10(-4)) and horses from 5 to 10 years old (P=0.018) resulted more infected in Bursa and Izmir provinces. Our findings indicate that R. equi infection actually occurs in all investigated provinces, suggesting the importance of serological survey to diagnose the infection and to prevent the zoonotic risk.
The production of cholesterol oxidase in two strains of Rhodococcus equi No. 23 from butter, and Rhodococcus sp. No. 33 from bacon, which had properties on biochemical and physiological tests almost similar to the strains of R. equi, was compared with that of the type strain (ATCC 6939) of R. equi. The intensity of cholesterol oxidase activity, both extracellular and membrane-bound, from the three strains was in the order No. 23, ATCC 6939 and No. 33. More extracellular enzyme was produced by strain No. 23 than by the other two strains. Halo formation on the agar medium containing cholesterol depended on the conversion of cholesterol to 4-cholesten-3-one by the extracellular cholesterol oxidase.
The negative mutant approach was used in this study to identify listerial cytolytic factors involved in cooperative haemolysis (CAMP-like phenomenon) with Staphylococcus aureus and Rhodococcus equi. A Listeria monocytogenes non-haemolytic mutant specifically impaired in listeriolysin O (LLO) production gave no CAMP reaction with S. aureus, and was virtually CAMP-negative with R. equi, indicating that the listerial sulphydryl-activated toxin played a major role in cooperative haemolysis. This was confirmed by direct evidence using purified LLO and alveolysin (from Bacillus alvei) in diffusion CAMP assays. To our knowledge, this is the first evidence of involvement of a sulphydryl-activated toxin in cooperative lytic processes. Phosphatidylcholine- and phosphatidylinositol-specific phospholipases C from L. monocytogenes did not seem to significantly contribute to cooperative haemolysis, as the corresponding mutants displayed wild-type CAMP reactions. In contrast, the sphingomyelinase C from Listeria iva-novii was the cytolytic factor responsible for the characteristic shovel-shaped CAMP reaction shown by this listerial species with R. equi. Possible mechanisms of lytic cooperation are discussed.
A 1-year-old neutered male domestic shorthair cat had an ulcerated, proliferative lesion in the submandibular area that did not respond to antibiotic therapy. Impression smears from the mass revealed septic pyogranulomatous inflammation, with large numbers of pleomorphic bacteria observed intracellularly within macrophages as well as neutrophils. Bacterial culture was consistent with a diagnosis of Rhodococcus equi, a facultative intracellular coccobacillus capable of replicating within macrophages. The cat's lesion resolved after treatment with rifampin and clarithromycin. R equi should be considered as a differential diagnosis when coccobacilli are recognized within macrophages in cytologic samples.
Clarithromycin offers numerous advantages over erythromycin and thus, is an attractive alternative for the treatment of Rhodococcus equi infections in foals. The disposition of clarithromycin was investigated in 6 foals after intragastric administration at a dose of 10 mg/kg body weight. Detectable serum concentrations of clarithromycin were found in 3 of 6 foals at 10 minutes and in all foals by 20 minutes post-administration. Time to peak serum concentration (Tmax) was 1.5 hours and peak serum concentration (Cmax) was 0.92+/-0.17 microg/ml. Mean serum concentrations decreased to 0.03 microg/ml at 24 h. No adverse reactions were noted during or after IG administration in any of the foals. Based on the pharmacokinetic parameters, the MIC90 of R. equi isolates, and predicted steady state concentrations, an oral dose of 7.5 mg/kg given every 12 hours would appear appropriate for the treatment of R. equi infections in foals.
Malakoplakia is an unusual inflammatory reaction to a variety of infections, characterized by the accumulation of macrophages containing the target-like calcospherites, the Michaelis-Gutmann body (MGB). We report three patients with acquired immunodeficiency syndrome with pulmonary malakoplakia associated with Rhodococcus equi infection; two patients were diagnosed at autopsy and one by examination of a transbronchial biopsy specimen. All three patients had pulmonary bacterial cultures and light and electron microscopic examination. The patients were 33-, 41-, and 43-year old men, human immunodeficiency virus-positive for 2, 6, 8 years, respectively. The two patients diagnosed at autopsy had cavitary lesions, and the patient diagnosed by biopsy specimen had nodular lesions on chest radiographs. Histologically, the lungs had well-circumscribed areas of infiltration with benign macrophages with granular cytoplasm, scattered MGBs, and numerous gram-positive coccobacilli. Electron microscopic examination showed intracellular coccobacilli, from 990 X 702 to 972 X 648 nm in diameter, with thick, homogenous cell walls, trilaminar cytoplasmic membranes, and dense cytoplasm with from one to five vacuoles. Electron microscopic studies showed that the bacteria within the pulmonary macrophages had thicker cell walls, less prominent nucleoid areas, and more vacuoles than the bacteria in cultures from the sputum and blood. The mature MGB ultrastructurally had a concentric, trilaminate structure with central mineralized core and was without recognizable bacterial forms. Early MGBs, however, consisted of a circular, electron-dense core containing bacteria, ultrastructurally similar to the R. equi seen in the culture. Pulmonary malakoplakia in patients with the acquired immunodeficiency syndrome might thus represent an acquired macrophage dysfunction of the intracellular digestion of phagocytized bacteria. The bacteria within the macrophages, however, seemed to have thicker cell walls compared with those in culture, and thus might be protected from enzyme digestion. It seems that MGBs are formed around the undigested bacteria as an alternative pathway for bacterial destruction, because R. equi was identified within the cores of early MGBs but not the mature or late stage MGBs.
Pulmonary infection by Rhodococcus equi is characterized by indolent infection in an immunocompromised host with a propensity to form cavitary lesions. Mortality can be greater than 50%; treatment involves prolonged therapy with multiple antibiotics and, occasionally, surgical resection. Recurrence is common. We report a case of a liver transplant patient with a pulmonary nodule caused by R equi; the nodule followed a benign clinical course and resolved spontaneously. This case illustrates that the spectrum of disease caused by R equi is not fully appreciated and that significant pitfalls complicate the diagnosis and management of infection by this unusual and probably underrecognized pathogen.
Rifampicin is a principal drug used to combat infections by mycobacteria and related organisms. Most strains of Mycobacterium are able to inactivate this antibiotic by ribosylation via an ADP-ribosylated intermediate. We found that this inactivation was inhibited by light at levels similar to those prevailing in laboratory environments. Rifampicin resistance arising from the cloned ADP-ribosyl transferase was also greatly diminished at these light levels. The cloned Rhodococcus equi monooxygenase which inactivates this antibiotic by a different mechanism was, in contrast, not inhibited by light.
Alveolar macrophages are almost invariably present in percutaneous fine-needle aspiration cytology of the lung. They may predominate, appear foamy and finely vesiculated, or may reflect the cellular composition of the lung mass lesion. In a review of 172 cases of "negative for malignant cells" from the percutaneous lung fine-needle aspiration cytology file in an 8-year period at Hines VA Hospital, the vacuolated macrophages were evaluated qualitatively and quantitatively. Among the 53 cases (34%) showing vacuolated macrophages, only 5-25% of the cells were multivesiculated, the cytoplasmic vacuoles were few, focal, and occasionally global, and the majority of the vacuolated macrophages contained anthracotic or hemosiderin pigments. One case exhibited striking multivesiculation in at least 95% of macrophages and also in bronchial and alveolar cells, fibroblasts, and endothelial and inflammatory cells, a finding consistent with amiodarone toxicity (index case 1). The diagnosis was confirmed on subsequent transbronchial lung biopsy. In another patient with clinical HIV infection, the multivesiculation was also seen in 95% of the macrophages with associated acute inflammatory exudate, coccobacilli, and a positive culture for Rhodococcus equi (index case 2). In most cases, the vacuolated macrophages are reactive and inflammatory. Occasionally, as in our index cases they may actually indicate a specific diagnosis.
The properties of azithromycin suggest that it may be an alternative to erythromycin for treatment of Rhodococcus equi pneumonia in foals. To investigate this possibility, the disposition of azithromycin in plasma, polymorphonuclear leukocytes (PMN), and alveolar cells was examined after a single administration in foals. Azithromycin suspension was administered orally (p.o.) at a dose of 10 mg/kg to five healthy 2-3-month-old foals. Two weeks later, azithromycin for injection was administered by intravenous (i.v.) infusion at a dose of 5 mg/kg to the same foals. Plasma samples were collected after p.o. and i.v. administration. Peripheral blood PMN and bronchoalveolar lavage fluid and alveolar cells were collected after p.o. administration. Azithromycin concentrations were determined by reverse-phase high-performance liquid chromatography (HPLC) with coulometric electrochemical detection. Azithromycin p.o. absorption was variable with a mean systemic availability of 39% (+/-20%). The plasma half-life was 16 and 18.3 h after i.v. and p.o. administration, respectively. Azithromycin had a very large volume of distribution (V(d)) of 11.6 L/kg [V(d(ss))] and 12.4 L/kg [V(d(area))]. The large V(d) can be attributed to high tissue and intracellular concentrations, exhibited by the high concentration of azithromycin in PMN and alveolar cells. The PMN half-life was 49.2 h. Dosage of 10 mg/kg of azithromycin p.o. once daily for foals with R. equi pneumonia is recommended for further study.
We report on a case with an unusual combination of pulmonary pathogens including Pneumocystis carinii, Rhodococcus equi and Toxoplasma gondii in an HIV infected patient. The multiple microbiological procedures required to ascertain the etiological diagnoses of pneumonitis in HIV-seropositive subjects are discussed.