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Enzyme-linked immunosorbent assay for Potomac horse fever disease.
An enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G (IgG) and IgM in natural and experimental infections of equids with Ehrlichia risticii was developed. Ehrlichial organisms purified from an infected mouse macrophage cell line were used as the antigen. IgM was separated from serum IgG by the expedient of spun-column chromatography, allowing the use of an indirect ELISA for quantitation of both IgG and IgM in the test sera. Among 16 paired sera from horses exhibiting clinical signs of Potomac horse fever, 8 were positive by the indirect fluorescent-antibody test (IFA), 11 were positive by the IgG ELISA, and 8 were positive by the IgM ELISA. All IFA-positive specimens were positive by the IgG ELISA, which appeared to be more sensitive than the IFA. In all cases, the IgG ELISA alone would have sufficed for diagnosis when acute- and convalescent-phase sera were available. When 26 single acute- or convalescent-phase serum samples were tested, the IFA detected 8, the IgG ELISA detected 10, and the IgM ELISA detected 6 positive serum specimens. The kinetics of IgG and IgM responses as determined by ELISA in two experimentally infected ponies which survived infection and challenges revealed that specific IgM was short-lived, falling to undetectable levels by day 60 postinoculation, whereas specific IgG persisted for more than 1 year. IgM and IgG were detected as early as days 1 and 10, respectively, postinoculation. The results suggest that the ELISA is more sensitive than the IFA and that the IgM ELISA may provide a means for early diagnosis of Potomac horse fever at or before the onset of clinical signs.
Laboratory diagnosis and characterization of renal disease in horses.
Laboratory evaluation of renal function in horses has advanced dramatically in the last 10 years largely as a result of the interest generated by the creative approach to diagnostic indices taken by Brobst, Traver, Coffman, and others. Some methods of assessing renal function discussed here are clearly outside the scope of a practice environment but are available in referral hospitals for use in difficult or unusual cases. Other methods described, such as calculation of fractional excretions and urine to serum creatinine ratios, are accessible and readily interpreted by the veterinary practitioner. These diagnostic methods should help in the early identification of renal disease in horses and therefore should allow veterinarians the option to treat renal disease before renal failure is advanced. In addition, practitioners may be able to offer more informed prognoses after a more complete evaluation of renal function.
An unusual incidence of neurological disease affecting horses during a drought.
The clinical, pathological and epidemiological factors were investigated in 12 horses presenting with severe neurological signs. Although the cases involved differing central (n = 1), spinal cord (n = 4) and peripheral nerve (n = 7) deficits in a number of instances, there were similar pathological findings. The possibility of a unifying aetiological factor, such as a toxicosis, is discussed because of the pathological similarities and as the cases appeared during an unusually long dry period.
[Corneal diseases in horses (part 2)].
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Studies on the physiopathology of chronic obstructive pulmonary disease in horses. I. Clinical signs.
Twenty cases of chronic cough originating in the lung and associated with loss of performance were clinically examined. The physical signs observed were compared with those observed in a control series of 38 clinically normal horses. Reduced work tolerance, coughing for more than 3 months and abnormal pulmonary sounds (râles) were primary signs of chronic obstructive pulmonary disease (COPD). Forced abdominal expiratory efforts and pumping of the anus were regarded as confirmatory signs. Neither nasal discharge nor increased marginal distance was found to be a reliable sign of COPD. The mean respiratory frequency of the COPD subjects, namely 25,4 per minute, was significantly higher than the 16,7 per minute (P less than 0,001) of the 38 normal subjects.
Sulfidoleukotriene generation from peripheral blood leukocytes of horses affected with insect bite dermal hypersensitivity.
Sulfidoleukotrienes (sLT) generated in vitro after incubation of equine peripheral blood leukocytes (PBL) with different inducing agents were determined in 18 healthy and 16 insect bite dermal hypersensitivity (IDH)-affected horses. PBL from these 32 horses were stimulated with Concanavalin A, Parascaris equorum, Culicoides nubeculosus and Simulium extracts, and with a six-Grass mix. The cells of all but four horses generated sLT after incubation with Concanavalin A; these four horses did also not produce sLT with the other inducing agents. Of the 28 remaining horses (12 affected with IDH and 16 healthy), all but three generated sLT with the P. equorum extract. The six-Grass mix did not induce sLT production in any of the tested horses. sLT generation with Concanavalin A and Parascaris was statistically not different between IDH-affected and healthy horses. PBL of the diseased horses, however, produced significantly more sLT with the Culicoides (p < 0.01) and Simulium (p < 0.05) extracts than those of the healthy animals. Additionally, sLT generation with the Culicoides extract was measured at different times of the year in one IDH-affected animal and remained high even in winter, when the horse was asymptomatic. sLT and histamine release were determined in 10 horses in parallel. Positive correlations of 0.81 and 0.82 for Concanavalin A and Parascaris (p < 0.01 and p < 0.05, respectively), and of 0.95 and 0.94 for Culicoides and Simulium (p < 0.01) were found between sLT and histamine release. These results indicate that, alike in humans, sLT are released in vitro from equine basophils along with histamine in response to various stimuli and that immediate type hypersensitivity reactions to Culicoides and Simulium are often involved in the pathogenesis of IDH. Thus, sLT generation from equine basophils offers an in vitro diagnostic tool for IDH even in sensitised but asymptomatic horses.
Equine intestinal clostridiosis. An acute disease in horses associated with high intestinal counts of Clostridium perfringens type A.
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Interleukin-4 and interferon-gamma gene expression in summer pasture-associated obstructive pulmonary disease affected horses.
We hypothesised that horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD) react to an allergen or allergens in their summer environment that is either absent or present at lower levels in their winter environment; and that such allergens stimulate SPAOPD-affected horses to produce a different T helper lymphocyte cytokine profile from that of control horses. The primary objective of this study was to determine the cytokine mRNA profile of T helper lymphocytes obtained from summer pasture-associated obstructive pulmonary disease (SPAOPD) affected horses when 1) the horses were showing signs of disease (summer) and 2) they were in clinical remission (winter). A further objective was to determine the differences between cytokine mRNA T helper lymphocyte profiles of control and affected horses in the summer and winter seasons. Interleukin 4 (IL-4) and interferon-gamma (IFN-gamma) mRNA expression levels were increased in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cell (PBMC) samples of affected horses during disease expression. No significant amounts of IL-5 mRNA were detected in any of the samples. These results suggest that there is an allergic component to SPAOPD of horses and that appropriate manipulation of the immune system could offer hope for treatment and prevention of the disease in the future. Further research studies will be needed to determine the most appropriate treatments to use to alter the antigen-stimulated cytokine profile being expressed by SPAOPD-affected horses or to alter the effects that these cytokines produce.
Trypanosoma evansi in inbred and Swiss-Webster mice: distinct aspects of pathogenesis.
Trypanosoma evansi (Trypanosomatidae, Kinetoplastida) is a salivarian trypanosomatid that infects eight mammal orders spread over America, Europe and Asia. In Brazil, T. evansi is the etiological agent of "Mal de Cadeiras", a horse disease very often described in the region known as Pantanal do Mato Grosso. Few data concerning the genetic diversity and biology of subpopulations of T. evansi that circulate in Brazil are available. The factors that modulate the interaction of this parasite with its hosts also remain to be elucidated. Here we evaluated the course of experimental infection of six T. evansi isolates derived from domestic and wild animals in Swiss-Webster mice and three Mus musculus lineages. The follow-up included biological, immunological as well as biochemical and hematological parameters. The same isolates as well as three others were characterized by pulsed-field electrophoresis. Our results showed that T. evansi isolates displayed significant differences regarding behavior and morbidity patterns in the distinct mouse lineages. Nevertheless, these differences could not be correlated with pulsed-field electrophoresis profiles. Indeed, concerning this molecular marker, only microheterogeneity was observed. Moreover, we observed that the outcome of the infection is defined by both host genetic background and peculiarities (virulence factors) of the distinct T. evansi isolates. Anemia and hypoglycemia were the only features that could be observed in all mouse lineages, independently of the inoculated T. evansi subpopulation. In addition, our data also show that Mus musculus is a suitable model host for the study of the different pathogenetic features of T. evansi infection.
Control of chronic obstructive pulmonary disease in the horse.
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Paracentesis as an aid to the diagnosis of abdominal disease in the horse.
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Equine inco-ordination and wobbler disease of young horses.
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Studies on the physiopathology of chronic obstructive pulmonary disease in the horse. VI. The alveolar dead space.
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Urine cartilage oligomeric matrix protein (COMP) measurement is useful in discriminating the osteoarthritic Thoroughbreds.
OBJECTIVE: To quantify the urinary concentration of cartilage oligomeric matrix protein (COMP), and to evaluate the relationship between urinary COMP concentration and the catabolic activity of synovial fluid (SF) in diseased horses. METHODS: COMP in horse urine was detected by immunoblotting with a monoclonal antibody (mAb; 14G4) raised against equine COMP from articular cartilage. Urine and serum samples were obtained from 83 Thoroughbred horses with aseptic joint diseases (AJD, 79 horses) or septic joint diseases (SJD, four horses) at the time of anesthesia induction, and samples of SF were obtained during surgery. Control samples of urine (n=111) were collected from normal horses free of any orthopedic diseases after they had been racing. COMP concentration was determined in all samples using inhibition enzyme-linked immunosorbent assay (ELISA) with mAb 14G4. SF samples were also used for the quantification of gelatinase activity. RESULTS: Positive bands of COMP fragments were determined on the immunoblots with mAb 14G4. The urinary COMP concentrations in AJD and SJD horses (1.02+/-0.75 and 1.55+/-1.17 microg/100mg creatinine, respectively) were significantly higher than normal (0.57+/-0.29 microg/100mg creatinine). In 55 horses with fractures in the AJD group there was a logarithmic relationship (r=-0.45, P<0.001) between the urinary and SF COMP measurements, while the urinary COMP level was positively correlated with matrix metalloproteinase (MMP)-2 and -9 activities (r=0.30, P<0.05 and r=0.51, P<0.001, respectively) in SF. CONCLUSIONS: The urinary COMP assay with mAb 14G4 is useful for discriminating horses with osteoarthritis. The higher COMP levels in urine from such horses would be indicative of enhanced proteolytic activity, in addition to the increased COMP levels in the diseased joints.
Inflammatory bowel disease in horses: 11 cases (1988-1998).
OBJECTIVE: To determine clinical and surgical abnormalities in, and long-term outcome of, horses that undergo surgery because of colic secondary to inflammatory bowel disease (IBD). DESIGN: Retrospective study. ANIMALS: 11 horses. PROCEDURE: Medical records of horses that had undergone abdominal surgery and in which IBD had been diagnosed on the basis of histologic examination of intestinal biopsy specimens were reviewed. RESULTS: 5 horses were examined because of acute colic and 6 were examined because of chronic colic. At surgery, all 11 horses had edematous or hemorrhagic bowel segments suggestive of IBD. In addition, 6 horses had circumferential mural bands (CMB) causing constriction of the small (4 horses) or large (2) intestine. Intestinal resections were performed in 7 horses. All 11 horses survived surgery and were discharged from the hospital; 10 horses were still alive at the time of follow-up (1.5 to 7 years after surgery). CLINICAL IMPLICATIONS: Results suggest that IBD is an uncommon cause of colic in horses. Surgical resection of segments of intestine with constrictive CMB may relieve clinical signs of colic. Horses with IBD that had surgery had a good prognosis for long-term survival.
Effect of vaccination against leptospirosis on the frequency, days to recurrence and progression of disease in horses with equine recurrent uveitis.
OBJECTIVE: To evaluate the effect of vaccination against leptospirosis on frequency and days to recurrence of uveitis and progression of disease in horses with equine recurrent uveitis (ERU). ANIMALS: Forty-one horses with ERU. PROCEDURE: Horses were randomly assigned to experimental (vaccinated) or control groups. Vaccine containing six serovars of Leptospira or placebo was administered, an ophthalmic examination performed and blood samples drawn on days 0, 28, 180 and 365. Antibody titers were measured against each serovar. Recurrence of uveitis was verified by ophthalmic examination. Results of the initial and final ophthalmic examinations were compared and progression of disease defined as an increase in extent of synechiae, or development of new or progression of an existing cataract. RESULTS: Vaccination increased the average geometric mean serum antibody titer from 225 on day 0, to 4077 and 593 on days 28 and 180, respectively. After the second vaccination, days to first recurrence was significantly longer (median 126 days; range 24-231 days) when compared with controls (median 86 days; range 14-192 days, P=0.04). Recurrence of ERU was observed among 7/20 (35%) vaccinated horses and 12/21 (57%) controls; however, this difference was not statistically significant (P=0.061, OR 0.25, 95% CI 0.06, 1.07). More horses in the experimental group 13/20 (65%) experienced progression of disease when compared with controls 12/21(57%); however, this difference was statistically nonsignificant (P=0.35). CONCLUSIONS: Vaccine significantly increased days to recurrence, but failed to slow the progression of disease. These data do not support the use of vaccination against leptospirosis as adjunct therapy for the routine treatment of horses with ERU.
Evaluation of pleural fluid in the diagnosis of thoracic disease in the horse.
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