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Rhodococcus equi (R. equi), a gram-positive facultative intracellular pathogen, is a common cause of pneumonia in foals and represents a major cause of disease and death. The aim of the present study was to investigate the time-depended changes in White Blood Cells (WBC), basophils (Baso), neutrophils (Neu), lymphocytes (Lymf), monocytes (Mon), eosinophils (Eos), platelet (PLT) counts, fibrinogen (Fbg) concentration, interferon (IFN-α, IFN-γ) and interleukins (IL-2 and IL-10) in foals with clinical R. equi pneumonia. The main treatment was with azithromycin-rifampicin for 14 days. Blood was sampled prior to, 7 and 14 days after starting therapy. Treatment was associated with significantly decreased counts of WBC, (25.6 ± 6.7 and 14.2 ± 2,7 × 10 3 /ml), Neu (18.6 ±6.2 and 10.7 ± 3.1 × 10 3 /ml), Mon (1.5 ± 0.5 and 0.9 ± 0.2 × 10 3 /ml) and Fbg (539 ± 124 and 287 ± 26 g/dl) between day 0 and day 14. IL-2 and IL-10 concentrations were significantly increased (P = 0.028, P = 0.013, respectively) after treatment, whereas IFN-α and IFN-γ concentrations were not. The diagnostic potentials of INF-α, INF-γ, IL-2 and IL-10 per se seems not very high, however, the study suggests that the activity change of selected interleukins in the course of the disease may be associated with amelioration. We concluded that patterns of serum concentration changes of INF-α, INF-γ, IL-2 and IL-10 may help in the study of the innate immune response in foals during infection and treatment of R. equi pneumonia., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

A Gram-stain-positive coccus was isolated from the blood of a paediatric patient suffering from gastroenteritis. The taxonomic position of this catalase-positive, non-motile, non-spore-forming facultative anaerobe designated as strain MKL-02 T was investigated using a polyphasic approach. Colonies grown on tryptic soy agar with 10 % sheep blood were circular, creamy yellow, and convex. Phylogenetic analysis based on 16S rRNA gene and whole-genome sequences revealed that this strain was most closely related to Arsenicicoccus bolidensis CCUG 47306 T within the cluster of the genus Arsenicicoccus . Average nucleotide identity and digital DNA-DNA hybridization values between strain MKL-02 T and A. bolidensis DSM 15745 T , A. dermatophillus DSM 25571 T and A. piscis DSM 22760 T were 89.5 and 37.0 %, 79.6 and 22.4 %, and 75.9 and 21.0 %, respectively. The genomic size of strain MKL-02 T was 3 423 857 bp with a 72.7 mol% G+C content. Growth was observed at 10-45 °C (optimum, 37-40 °C) and pH 6.0-10.0 (optimum, pH 7.0), in the presence of 0-10 % (w/v) NaCl (optimum, 0.5 %). Cells of strain MKL-02 T were non-motile cocci and 0.50-0.60 µm long, as determined by transmission electron microscopy. The strain was catalase-positive and oxidase-negative. The major fatty acid type (>10 % of total) was C 15 : 0 . The polar lipid profile consisted of two unidentified phospholipids, three unidentified lipids and an unidentified aminophospholipid. The strain contained MK-8 (H 4 ) as the predominant menaquinone. Based on phylogenetic and phenotypic considerations, it is proposed that strain MKL-02 T be classified as a new species, named Arsenicicoccus cauae sp. nov. The type strain is MKL-02 T (=NCCP 16967 T =JCM 34624 T ).

Rhodococcus equi is a prevalent cause of pneumonia in foals worldwide. Our laboratory has demonstrated that vaccination against the surface polysaccharide β-1→6-poly- N -acetylglucosamine (PNAG) protects foals against intrabronchial infection with R. equi when challenged at age 28 days. However, it is important that the efficacy of this vaccine be evaluated in foals when they are infected at an earlier age, because foals are naturally exposed to virulent R. equi in their environment from birth and because susceptibility is inversely related to age in foals. Using a randomized, blind experimental design, we evaluated whether maternal vaccination against PNAG protected foals against intrabronchial infection with R. equi 6 days after birth. Vaccination of mares per se did not significantly reduce the incidence of pneumonia in foals; however, activities of antibody against PNAG or for deposition of complement component 1q onto PNAG was significantly ( P  < 0.05) higher among foals that did not develop pneumonia than among foals that developed pneumonia. Results differed between years, with evidence of protection during 2018 but not 2020. In the absence of a licensed vaccine, further evaluation of the PNAG vaccine is warranted, including efforts to optimize the formulation and dose of this vaccine. IMPORTANCE Pneumonia caused by R. equi is an important cause of disease and death in foals worldwide for which a licensed vaccine is lacking. Foals are exposed to R. equi in their environment from birth, and they appear to be infected soon after parturition at an age when innate and adaptive immune responses are diminished. Results of this study indicate that higher activity of antibodies recognizing PNAG was associated with protection against R. equi pneumonia, indicating the need for further optimization of maternal vaccination against PNAG to protect foals against R. equi pneumonia.

Background: Cellulosimicrobium cellulans is a gram-positive filamentous bacterium found primarily in soil and sewage that rarely causes human infection, especially in previously healthy adults, but when it does, it often indicates a poor prognosis., Case Presentation: We report a case of endocarditis and intracranial infection caused by C. cellulans in a 52-year-old woman with normal immune function and no implants in vivo. The patient started with a febrile headache that progressed to impaired consciousness after 20 days, and she finally died after treatment with vancomycin combined with rifampicin. C. cellulans was isolated from her blood cultures for 3 consecutive days after her admission; however, there was only evidence of C. cellulans sequences for two samples in the second-generation sequencing data generated from her peripheral blood, which were ignored by the technicians. No C. cellulans bands were detected in her cerebrospinal fluid by second-generation sequencing., Conclusions: Second-generation sequencing seems to have limitations for certain specific strains of bacteria.

An 83-year-old previously self-sufficient man was referred to our hospital for a fever, severe tenderness over the lumbar spine, and elevated C-reactive protein levels. Computed tomography revealed fluid collection in the intervertebral space of L3/4. Gram-positive, short rod-shaped bacteria were isolated from two sets of blood cultures. A 16S rRNA sequence analysis of an isolate showed a similarity of 98.1% to the nearest type strain Brachybacterium squillarum JCM 16464 T . Biochemical characteristics of the presently isolated strain differed from those of the most closely related species of the genus Brachybacterium. The patient was successfully discharged on day 73 of admission with antimicrobial therapies and showed no recurrence during outpatient visits. Brachybacterium spp. have mainly been isolated from the environment, and human Brachybacterium infections have rarely been documented to date. To our knowledge, this is the first clinical isolation of Brachybacterium sp. as a causative pathogen of bloodstream infection., (Copyright © 2018. Published by Elsevier Ltd.)

Reasons for Performing Study: Rhodococcus equi (Rhodococcus hoagii/Prescottella equi) is a common cause of foal pneumonia, but its diagnosis remains a challenge for equine veterinarians. While the VapA-specific (virulence-associated protein A) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) has low sensitivity and specificity for detecting pneumonic foals, little is known about VapA-specific IgG subclasses., Objectives: To evaluate the performance of VapA-specific ELISA for IgG and its subclasses IgGa, IgGb and IgG(T) in the early diagnosis of pneumonia caused by R. equi., Study Design: Assay validation followed by assessment of diagnostic performance using archived samples from animals of known status., Methods: Serum samples from exposed (n = 125) and nonexposed adult horses (n = 10) and from experimentally challenged and naturally infected foals were used for ELISA validation. Post mortem and tissue culture records of the last 24 years from the Institute for Experimental Pathology at the University of Iceland in Keldur, Iceland laboratory were evaluated to confirm the absence of R. equi cases in Iceland. The diagnostic performance of VapA-specific IgG and its subclasses was evaluated using banked serum samples from pneumonic (n = 21) and healthy foals (n = 80). To evaluate each IgG assay, a cut-off value was selected based on receiver operating characteristic curve analysis and used to calculate sensitivity and specificity. The intra- and interassay coefficients of variation were calculated for each ELISA., Results: Using sera from Iceland, where R. equi infection has not been reported, the VapA-specific IgG ELISA differentiated exposed from nonexposed horses. When used to identify infected foals, VapA-specific IgG, IgGa and IgGb had no diagnostic value. In contrast, IgG(T) had high sensitivity and specificity., Conclusions: Horses from Iceland are not exposed to VapA(+) R. equi and can serve as negative controls. VapA-specific IgG subclasses, with the exception of IgG(T), are poor predictors of disease. Further investigation on the use of IgG(T) as a diagnostic tool in field conditions is needed., (© 2015 EVJ Ltd.)

Early diagnosis and prevention of Rhodococcus equi pneumonia in foals represent important goals for equine clinicians. Recent protocols for diagnosis and treatment of Rhodococcosis in foals typically rely on a multimodal approach based on sonographic evidence suggestive of pyogranulomas, sonographic abscess scores and laboratory findings including plasma fibrinogen concentrations, blood biochemistry testing and platelet and leukocyte counts. The aim of this study was to assess the utility of weekly testing of serum amyloid A (SAA) and plasma fibrinogen concentrations in foals to achieve early diagnosis of R. equi pneumonia prior to the onset of clinical signs. This testing was used to simulate a clinically practical screening procedure and compared with thoracic ultrasonography performed in parallel. The present study suggests that SAA does not represent a reliable early marker of Rhodococcosis when plasma concentrations are tested weekly. However, when clinical signs of R. equi pneumonia are present, SAA concentrations may allow clinicians to obtain 'real-time' indications concerning both the progress of infection and the effectiveness of therapy. This study raises the possibility that plasma fibrinogen monitoring starting at 1 week of age and repeated on a weekly basis, could serve as a screening test allowing clinicians to identify foals as suspected of R. equi infection. Future investigations regarding both physiological plasma fibrinogen concentrations in foals as well as fibrinogen kinetics in foals affected with R. equi pneumonia, including the establishment of appropriate reference intervals for the test method employed in this study, will be necessary in order to clarify this possibility., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Rhodococcus equi infection occurs worldwide and is one of the major causes of losing foals in the first six months of life. The application of serological tests in the diagnostics of rhodococcosis is limited, however they play a crucial role in immunological studies. The objective of this study was to develop and standardize ELISA test for the determination of the level of antibodies against Rhodococcus equi in equine serum and colostrum.Bacterial cell lysate was used as antigen. The test was standardized on 175 sera obtained from adult horses kept on rhodococcosis-free and endemic farms. Positive and negative control sera were used. The test detected IgG antibodies mainly against VapA protein, which was confirmed by Western blot analysis. The test was easy to perform, did not require inactivation of sera and had low well-to-well variation. The shelf life of antigen-coated ELISA plates was 21 days.The test allowed to reveal significant increase of R. equi-specific antibodies in both serum and colostrum in response to the vaccination (p<0.001). Therefore it can be applied to the evaluation of efficacy of immunization. Moreover, no statistically significant difference in the baseline antibody level in adult horses from rhodococcosis-free and endemic farm was revealed (α=0.05)., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The purpose of this study was to compare the clinical, microbiological, radiological, haematological and cytological features of foals with pneumonia caused by Rhodococcus equi infection and with other bacteria, in order to provide markers for early diagnosis and treatment. A retrospective study of 113 cases of bacterial pneumonia was undertaken. Although there was considerable overlap in the affected populations, foals with R. equi pneumonia were significantly younger and had higher respiratory rates. Radiological evidence of thoracic abscessation had a sensitivity of 71% and a specificity of 85% for the diagnosis of R. equi pneumonia. Foals positive for R. equi also had higher peripheral white cell counts and fibrinogen concentrations than animals not infected with this pathogen. Respiratory rate, fibrinogen concentration and the log of the neutrophil count were retained in the final multivariate analysis. Using microbiological culture as the 'gold standard', identification of Gram-positive coccobacilli in tracheal aspirates was highly specific (91%), but poorly sensitive (35%) for R. equi infection. White cell counts >20,000cells/μL (86% specificity), fibrinogen concentrations >700mg/dL (92% specificity), radiological evidence of thoracic abscessation (85% specificity), and the presence of Gram-positive coccobacilli in tracheal aspirates (91% specificity) in pneumonic foals are highly suggestive of R. equi infection and justify early targeted antimicrobial intervention while awaiting culture results., (2009 Elsevier Ltd. All rights reserved.)

In this study, the authors evaluated parenteral treatment of zebu cattle, with naturally and experimentally induced bovine dermatophilosis, in western Sudan, using four different antibiotic treatments. In terms of recovery rate, weight gain, avoiding relapse and preventing death, gentamycin was found to be the most effective treatment, followed by a combination of penicillin and streptomycin and, finally, long-acting oxytetracycline. However, enrofloxacin was not successful. A significant improvement in the red blood cell count was noticed among cattle treated with penicillin-streptomycin (p = 0.021) and gentamycin (p = 0.029). All treated cattle, except those treated with enrofloxacin, showed a significant improvement in mean corpuscular haemoglobin concentration (p = 0.021); mean corpuscular volume (p = 0.021), and white blood cell count (p < 0.021). Significant improvements were observed among treated cattle in their total levels of protein, calcium (p = 0.021) and cholesterol (p < 0.05), when compared to untreated cattle infected with Dermatophilus congolensis. This study recommends gentamycin as a drug of choice for the parenteral treatment of dermatophilosis. Treatment was not only effective in early, mild cases but also useful among moderately and heavily affected cattle. According to the observations of the authors, when no intervention took place, the condition of moderately and heavily affected cattle deteriorated and/or resulted in death.

Tsukamurellae are strictly aerobic Gram-positive rods that can be easily misidentified as Corynebacterium species, Rhodococcus species, Nocardia species, Mycobacterium species, or other Gram-positive aerobic rods. They have been uncommonly reported as a cause of different human infections, including bloodstream infections. We describe 2 new cases of catheter-related bloodstream infections (CR-BSI) caused by Tsukamurella species and review 12 similar cases reported in the literature. Conventional procedures have often misidentified Tsukamurella species as other aerobic Gram-positive rods. This misidentification could be avoided using genotyping. All cases ultimately required the withdrawal of the infected line. The literature provides no firm conclusions regarding ideal choice or duration of antimicrobial therapy for this infection. Tsukamurella species should be added to the list of agents able to produce CR-BSI. Genotypic methods such as PCR 16S rRNA can allow a reliable identification at the genus level of Tsukamurella strains faster than a combination of conventional phenotypic methods.

Reasons for Performing Study: Prognosis of Rhodococcus equi pneumonia can be challenging because the course of the disease is often insidious and overt clinical signs are subtle. Early diagnosis is considered desirable because it may offer the chance of more successful implementation of treatment and, thereby, improved outcome. Serological tests have previously failed to be accurate for early detection or diagnosis. Measurement of serum amyloid A (SAA) prior to and at the time of clinical signs was therefore chosen in order to assess its potential clinical use., Objective: To determine whether SAA concentrations differentiate foals affected with R. equi pneumonia from unaffected foals, either prior to the onset of disease or at the time of onset of clinical signs., Hypothesis: SAA concentrations are significantly higher among foals that develop R. equi pneumonia than in foals from the same environment that remain clinically unaffected., Methods: Serum samples were obtained from 212 foals 7-14 days and 196 foals 21-28 days post partum, and from affected foals and age-matched controls at the time of onset of signs of pneumonia. SAA concentration was determined for each sample., Results: There were no significant differences between SAA concentrations of foals with R. equi and clinically unaffected foals during the 2 periods of examination or at the time of onset of clinical signs of R. equi pneumonia., Conclusions: Concentrations of SAA are variable among foals with R. equi pneumonia and cannot be used reliably either as an ancillary diagnostic tool or to screen for early detection of disease during the first month post partum., Potential Relevance: Bimonthly monitoring concentration of SAA is not useful as a screening test for early detection of R. equi pneumonia and does not facilitate diagnosis of this disease when used according to the protocol of this study.

Objective: To evaluate WBC concentration, plasma fibrinogen concentration, and an agar gel immunodiffusion (AGID) test for early identification of Rhodococcus equi-infected foals., Design: Prospective study., Animals: 162 foals from a farm with enzootic R equi infection., Procedure: Blood samples were obtained from each foal at 4-week intervals for measurement of WBC and plasma fibrinogen concentrations and at 2-week intervals for detection of anti-R equi antibody by an AGID assay. Diagnostic performance of WBC and fibrinogen concentrations was assessed by use of receiver operating characteristic curve analysis. For each assay, sensitivity, specificity, and predictive values were calculated at various cutoff points; bacteriologic culture of R equi from a tracheobronchial aspirate was used as the reference standard test., Results: Diagnostic performance of WBC concentration was significantly higher than that of fibrinogen concentration. Sensitivity and specificity of measurement of WBC concentration at a cutoff of 13,000 cells/microL were 95.2 and 61.2%, respectively; at a cutoff of 15,000 cells/microL, sensitivity was 78.6% and specificity was 90.8%. When a positive test result was used as the cutoff, sensitivity of the AGID assay was 62.5% and specificity was 53.8%., Conclusion and Clinical Relevance: Monitoring WBC concentration is a useful approach for early detection of infected foals on farms with a high prevalence of R equi pneumonia. In contrast, serologic surveillance by use of an AGID assay is of little benefit for that purpose.

It has been suggested that in cases of puerperal endometritis of cattle infected with Escherichia coli and Arcanobacterium pyogenes, the neutrophils are compromised in their defense capacity or downregulated functionally. In addition to direct bacterial effects, contents of lochial secretions and secreted products of locally activated polymorphonuclear neutrophilic granulocytes (PMNs) may also account for changes in function of freshly immigrating neutrophils. In this study, lochial secretions were obtained from healthy cows and from cows infected by E. coli or A. pyogenes. Separated uterine PMN of infected cows displayed an altered phenotype and function which correlated with the degree of bacterial contamination. Concurrently tested circulating PMN showed no such changes. Infected lochial secretions sterilized by filtration also changed the phenotype of blood PMN. Lochial secretions of healthy cows displayed only minor effects. The effects on PMN function in infected cows varied: ingestion was less affected, whereas generation of reactive oxygen species (ROS) was severely depressed. Concurrently tested purified bacterial products (solubles and fragments) of E. coli and A. pyogenes did not induce the phenotypical and functional changes observed in blood PMN. Since infected lochia also contained high numbers of immigrated and probably activated PMN, the influence of supernatants from phorbol myristate acetate-activated PMN were tested on freshly isolated blood PMN. Such supernatants also increased the expression of certain surface molecules and inhibited the ROS generation. Thus, reduced function and altered phenotypes of PMN which immigrate into the uteri of cows with bacterial endometritis is due not only to interactions with bacteria or bacterial products, but is also to the uterine milieu.

In this study, the effects of orally administered viable Lactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined. For this purpose, we used the Trichinella spiralis host resistance model. Two weeks before and during T. spiralis infection, rats were fed 10(9) viable L. casei bacteria 5 days per week. The T. spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L. casei. In both rat strains fed L. casei, serum T. spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased. In the model, no significant effects of L. casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed. Serum specific antibody responses, other than IgG2b, were not changed by feeding of L. casei. In contrast to L. casei, it was shown that orally administered Bifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model. Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L. casei. Furthermore, our data do not indicate that the effect of oral supplementation with L. casei is dependent on the genetic background of the host.

After isolating the two virulent strains of Rhodococcus equi from Alpaca, a serological survey of Rhodococcus equi infection was carried out on 57 blood samples of Alpaca collected in Central Italy. The survey was performed with an ELISA test using a reference R. equi strain as antigen (ATCC 33701). Four (7.0%) sera (OD greater or equal to 0.3) tested positive, while five (8.77%) were considered doubtful (OD between 0.2 and 0.3). This is the first serological survey of R. equi infection in Alpaca in Italy. The results indicate that besides the horse R. equi infection could also affect some ruminant species. The ELISA test was recently introduced as a reliable diagnostic method in horses and was adapted to alpaca.

A non-competitive chemiluminescence enzyme immunoassay for measuring serum amyloid A (SAA) in equine serum was developed. A polyclonal anti-equine-amyloid A antiserum specific for equine SAA was utilized, and the assay was standardized using highly purified equine SAA. An acute phase horse serum was calibrated against the purified SAA and was used as standard when running the assay. Serum SAA concentrations in the range of 3-1210 mg/l could be measured. The reference range of SAA in clinically healthy adult horses was <7 mg/l. The clinical validation of the assay comprised the SAA responses after surgery and experimentally induced aseptic arthritis, and those associated with viral and bacterial infections. The SAA response after surgery (castration) was consistent, with peak concentrations on day 2 and a return to normal SAA concentrations within eight days. The aseptic arthritis produced an SAA response with a pattern similar to that seen after surgery, with peak concentrations of SAA 36-48 h after induction. Seven horses showed a biphasic pattern, with a second rise in SAA concentrations on day 4 and 5. All animals had SAA levels <7 mg/l on day 15. All horses with viral and bacterial infections had SAA concentrations above 7 mg/l. The ranges of SAA concentrations following the different types of inflammation overlap, being consistent with the unspecific nature of the SAA response. This study revealed that SAA is a sensitive and unspecific marker for inflammation, and describes the dynamics of the SAA response after standardized and well defined tissue damage.

Objective: To evaluate the ability of nucleic acid amplification techniques to detect Rhodococcus equi in equine buffy coat, blood, and tracheal wash fluid and to differentiate between virulent and avirulent strains of the bacteria., Sample Population: Blood anticoagulated with EDTA and tracheal wash fluid from healthy horses., Procedure: Logarithmic dilutions of virulent and avirulent strains of R equi were added to equine buffy coat and tracheal wash fluid samples. The DNA was extracted and amplified by polymerase chain reaction (PCR), using primers specific for the 16S ribosomal subunit gene and the virulence plasmid of R equi., Results: PCR with 16S ribosomal subunit primers amplified a 441-bp segment of DNA from virulent and avirulent strains of R equi, but not from samples containing other species of bacteria. The virulence plasmid primers amplified an 875-bp segment of DNA from virulent strains of R equi, but not from avirulent R equi, or from other species of bacteria. Virulent strains of R equi could be identified by PCR and differentiated from avirulent strains within 12 to 24 hours after sample collection, with as few as 10 to 100 organisms present., Conclusions: PCR can be used to rapidly and accurately identify R equi in equine blood and tracheal wash fluid samples and can differentiate between virulent and avirulent strains of the organism., Clinical Relevance: Because PCR can confirm a diagnosis of R equi infection in horses more rapidly and specifically than use of standard culture techniques, extrapolation of this assay to soil and fecal samples could be useful in epidemiologic studies and studies of environmental disinfection or decontamination.

Whole blood and serum were collected from foals to determine the prevalence of Equine herpesvirus type 2 (EHV 2) infection in foals, age at which infection can first be identified and serological responses to infection. Equine herpesvirus type 2 was isolated from peripheral blood mononuclear cells (PBMC) from 68 of 69 foals, 1-8-months-old, sampled once. Virus isolation was performed twice at intervals of 2-7 months on PBMCs from 33 foals and EHV2 was isolated on both occasions in all but one foal (negative, then positive). Regression analysis of log2-transformed reciprocal serum EHV2 virus neutralising (VN) titres revealed that in foals age 1-7 months, EHV2 VN antibody titre was positively correlated with age (r = 0.94). Paired serum samples were obtained from 58 foals, with the first samples collected age 1-6 months and the second samples collected 2-4 months later. There were significant (P < 0.05) increases in mean VN titres to EHV2 in foals sampled initially at age 1-4. Eight foals had blood sampled prior to sucking and at age 7, 20, 30 and 45 days. Each foal was negative for EHV2 in PBMC and each foal had a negative serum EHV2 VN titre immediately after birth. Each foal was positive for EHV2 in PBMC by age 45 days, with the earliest isolation at 25 days. Tracheal aspirate fluid and peripheral blood were collected from 20 foals without clinical signs of respiratory disease and from 30 foals with clinical signs of lower respiratory disease. In 20 foals without clinical signs of respiratory disease, EHV2 was isolated from tracheal aspirates (1/20 foals) and PBMC (20/20 foals) and in 30 foals with such clinical signs, from trachea aspirates (20/30 foals: P < 0.01) and from PBMC (30/30 foals). In one 6-month-old foal, EHV1, but not EHV2, was isolated from the tracheal aspirate, 3 months after EHV2 had been isolated from a tracheal aspirate. These results demonstrate a greater prevalence of EHV2 in lower respiratory secretions in foals with clinically apparent lower respiratory disease, but a cause and effect relationship between the virus and lower respiratory disease remains to be elucidated. It is noteworthy, however, that of virus isolations performed on 50 tracheal aspirates, a virus (EHV1) other than EHV2 was isolated only once.

Over nearly two decades, 13 yellow- or orange-pigmented, fermentative gram-positive rods belonging to the genus Microbacterium were encountered in clinical specimens. All 13 strains, 10 of which came from blood cultures, were initially identified as CDC coryneform group A-4 and A-5 bacteria according to the scheme of Hollis and Weaver for the identification of gram-positive rods. The clinical isolates were compared with the type strains of the six species constituting the genus Microbacterium as well as with three Microbacterium strains isolated from hospital environments. By biochemical methods only 5 of 13 clinical isolates could be identified to species level. Peptidoglycan analysis proved to be a valuable tool for differentiation between Microbacterium spp. and related genera, whereas cellular fatty acid analysis did not allow species identification within the genus Microbacterium. The 22 Microbacterium strains studied were, in general, susceptible to antimicrobial agents used in the treatment of infections caused by gram-positive rods. This report is the first one concerning the isolation of Microbacterium strains from clinical specimens. The sources as well as the mode of transmission remain to be established.

Polymorphonuclear neutrophil leukocytes (PMNL) from 8 healthy foals (2-14 weeks of age) and 2 foals with bacterial pneumonia were separated from whole blood using a 2 step Percoll gradient. Purified PMNL were tested for bactericidal function against Rhodococcus equi and Staphylococcus aureus in the presence of normal horse serum. The percentage uptake after a 15-min pre-incubation of PMNL and bacteria was also calculated. Ultrastructural examination of the interaction of R. equi and normal foal PMNL was performed after 15 min incubation. Results indicated that foal PMNL effectively phagocytose and destroy R. equi and S. aureus in the presence of normal horse serum. The mean percent uptake for R. equi was 99.3 +/- 0.4% and for S. aureus 99.9 +/- 0.1%. Further, 97.8 +/- 0.1% ingested R. equi and 98.4 +/- 0.1% ingested S. aureus were destroyed in the 15-min incubation period. Over the 3-h incubation, 91.9% of remaining R. equi were killed, but only 49.2 +/- 31.9% of S. aureus (P less than 0.01). Total bactericidal effect of foal PMNL, however, was 99.3 +/- 0.4% against R. equi and 99.9 +/- 0.1% against S. aureus. The percentage uptake and total bactericidal efficacy of neutrophils from sick foals was greater than 95%. Ultrastructural examination of the PMNL-R. equi interaction after 15 min incubation revealed phagocytosis of the bacteria and morphologic changes consistent with neutrophil degranulation. This study suggests that a defect in PMNL bactericidal capability is not likely to be a contributing factor in the pathogenesis of R. equi pneumonia in foals.

Pyogranulomatous pneumonia was induced in Thoroughbred foals by intranasal challenge with freeze-dried cultures of Rhodococcus equi (previously Corynebacterium equi). The incubation period was about 18 days and clinical signs were not seen for a further week. There were marked seasonal and individual foal differences in responses to infection. Elevations in serum caeruloplasmin oxidase activity and copper concentrations appeared to be sensitive indicators of infection. Serum zinc concentrations and serum alpha-mannosidase and alkaline phosphatase activities fell in the more severely infected foals. Use of trace elements and trace element-related parameters along with faecal culture for R. equi could prove useful for early diagnosis of field cases.