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Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus.

Equine infectious anemia virus (EIAV) is a lentivirus that causes persistent infections in horses. We hypothesized that high-avidity CTL specific for nonvariable epitopes might be associated with low viral load and minimal disease in EIAV-infected horses. To test this hypothesis, memory CTL (CTLm) responses were analyzed in two infected horses with high plasma viral loads and recurrent disease (progressors), and in two infected horses with low-to-undetectable viral loads and mild disease (nonprogressors). High-avidity CTLm in one progressor recognized an envelope gp90 epitope, and the data documented for the first time in EIAV that viral variation led to CTL escape. Each of the nonprogressors had high-to-moderate avidity CTLm directed against epitopes within Rev, including the nuclear export and nuclear localization domains. These results suggested that the epitope specificity of high- and moderate-avidity CTLm was an important determinant for disease outcome in the EIAV-infected horses examined.

Amino Acid Sequence↗

Concentration and molecular weight distribution of hyaluronate in synovial fluid from clinically normal horses and horses with diseased joints.

High molecular weight (MW) hyaluronate (HA) is an integral part of synovial fluid (SF), regulating many important physiologic and pathophysiologic mechanisms. Many of its effects depend on, or are reflected in, the concentration and MW of HA. High-performance liquid chromatography was used to assess simultaneously the concentration and MW of HA in SF obtained from horses with various arthritides: acute traumatic arthritis; chronic traumatic arthritis, including degenerative joint disease (DJD); and infectious arthritis. The size-exclusion column was calibrated, using appropriate HA concentration and MW standards, before the high-performance liquid chromatographic assays of the SF samples. Calibration of the column disclosed that the maximal limit for MW estimation of HA was around 3 million. In control joints, MW of HA ranged from 2 to 3 x 10(6) (mean 2.5 x 10(6)) and did not differ significantly from MW of HA in SF from horses with acute or chronic traumatic arthritis (mean 2 x 10(6); range 1.5 to 3 x 10(6)). Interestingly, a small amount of HA of moderately high MW (approx 1 to 1.5 x 10(6)) was detected in chromatograms of SF from infected joints. This degree of polymerization of SF HA was significantly (P < 0.01) lower, compared with that for control joints. There was no difference in mean (+/- SD) concentration of HA between control joints and joints with acute or chronic traumatic arthritis (0.33 +/- 0.12 g/L vs 0.18 +/- 0.03 g/L or 0.23 +/- 0.12 g/L), indicating that SF HA concentration probably should not be used as a diagnostic marker for the condition.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗

Causes of colic and types requiring surgical intervention.

The migration of strongyle larvae is the most common or basic underlying cause of colic in the horse. Disease conditions producing symptoms of colic occur in all sections of the intestinal tract and consist of impactions, torsions, herniations and foreign bodies. Colic also occurs as a result of pre- and post-partum diseases such as torsion of the uterus, haemorrhage, rupture and inversion of the uterus. In general, lesions resulting in circulatory obstruction are the types requiring surgical intervention. There are six general types of small intestine obstruction that lend themselves to surgical treatment; namely, volvulus, herniations, intussusceptions, stenosis of the lumen of the bowel by external bands or by foreign bodies and chronic inflammatory lesions. The large intestine is not subject to the variety of obstructive lesions found in the small intestine. Impactions count for a large percentage of the obstructions seen. A standing laparotomy for diagnostic purposes may be indicated. Small colon impactions are readily treated by standing laparotomy. Enteroliths are of very common occurrence in some areas of the country; they often result in rupture of the colon. Torsions of the colon produce septic shock very rapidly. The left dorsal colon moving medially or laterally and ventrally initiates the torsion. Clockwise rotation is most common. Massive intravenous therapy is needed to maintain hydration. Ventral midline laparotomy gives best access. Surgery must be performed very early to avoid massive tissue necrosis. Survival rate is 30 percent or less. The small colon is also capable of rotation and volvulus, and of strangulation in the umbilical or inguinal ring.

Animals↗

Population pharmacokinetics of marbofloxacin in horses: preliminary analysis.

Population pharmacokinetic of marbofloxacin was investigated on 21 healthy and 16 diseased horses to assess interindividual variability of drug exposure. Demographic, physiologic and disease covariables were tested using mixed effects models. As a preliminary analysis, this study has demonstrated that none of the tested covariables were significant in regression models for compartmental volumes or clearance of distribution, but the clinical status of the horse (healthy/diseased) was a significant covariable (P < 0.01) for systemic clearance. Clearance had a lower mean and a higher variance for diseased horses than healthy horses, with respectively a mean of 0.209 and 0.284 L/h/kg and a coefficient of variation of 52 and 15%. Consequently, variability of AUC was greater in diseased horses. Considering an AUC/MIC ratio below 60 h as a prediction of poor efficacy, a dosage regimen of 2 mg/kg intravenous was deemed to be inadequate for 19% of diseased horses if the MIC of the bacteria was 0.1 microg/mL. However 93% of diseased horses could achieve a ratio above 125 h, predicting a very good efficacy, for the MIC(90) of Enterobacteriacae (0.027 microg/mL).

Animals↗

Serum antiproteases and respiratory diseases of the horse.

The serum antiprotease (AAT) levels are reported in healthy horses and horses with respiratory diseases. Of the methods used, only the STIC test seemed to give useful results; this test showed variations in horses with respiratory diseases, especially in horses with acute alveolar pulmonary emphysema.

Alpha-Globulins↗

Serum level of cartilage oligomeric matrix protein (COMP) in equine osteoarthritis.

This study was designed to assay and compare cartilage oligomeric matrix protein (COMP) in horse sera, in samples from normal and joint diseased horses, and to investigate the relationships between COMP in sera and synovial fluids (SF) with keratan sulphate (KS) data. Sera from 38 horses free of any joint pathology (controls) and from horses with aseptic joint disease (AJD horses, n = 40) were assayed for COMP and KS concentrations. Of the 78 horses in the study, 53 were also assayed for COMP and KS concentrations in SF. COMP and KS were measured by inhibition ELISA, using monoclonal antibodies 12C4 and 5D4, respectively. The COMP concentration in sera from AJD horses (mean +/- s.d. 10.7 +/- 7.4 microg/ml) was significantly (P<0.02) lower than in control sera (14.8 +/- 7.8 microg/ml). The joint disease sera also had significantly lower (P<0.01) KS levels (180.5 +/- 61.8 ng/ml) than controls (237.1 +/- 116.1 ng/ml). A significant correlation (r = 0.52, n = 53, P<0.001) was seen between serum and SF in COMP levels; no such relationship was seen in KS levels. It is possible that serum COMP concentration could be a more specific marker of equine joint disease than any other described to date.

Animals↗

Myopathy in horses with pituitary pars intermedia dysfunction (Cushing's disease).

Fifteen horses with pituitary pars intermedia dysfunction were studied. The horses were of various breeds and between 15 and 28 years of age. Control horses matched for breed and age were studied for comparison. Evaluations included complete blood cell count and serum biochemical analysis, electromyography, and gluteus medius muscle biopsies for histochemical, morphometric, and ultrastructural analysis. No differences were found between groups of horses on routine laboratory analysis or electromyography. We demonstrated that muscle wasting in diseased horses was the result of atrophy of types 2A and 2B muscle fibers and loss of type 2B myofibers. Mild non-specific non-inflammatory myopathic alterations such as myofiber size variation, internal nuclei, perimysial, endomysial and sarcoplasmic fat accumulation were observed. At the ultrastructural level, subsarcolemmal mitochondrial accumulation and increased lipid droplets were evident. Similar to other species, this study confirmed atrophy of type 2 fibers as the cause of muscle mass loss in horses with Cushing's disease.

Animals↗

Assessment of radiographic positioning for the diagnosis of navicular disease in the horse.

Three of the standard radiographic views of the navicular bone were assessed in normal horses and horses with navicular disease to determine the most effective radiographic positioning. Using the upright pedal view, a pastern angulation of 20 degrees from the vertical produced the optimum result when radiographs were taken using a grid. The best result using the high coronary view was obtained with an anode-film distance of 1 1000 mm, and a tube-head angle of 50 degrees from the vertical. When the special navicular view described by Morgan was examined, the most satisfactory projection was obtained using an anode-film distance of 900 mm, a tube-head angle of 55 degree and the primary beam centred between the bulbs of the heel, for the diagnosis of navicular disease, a combination of the upright pedal view and the special navicular view appears most effective.

Animals↗

Diagnostic methods applied to analysis of an outbreak of equine influenza in a riding school in which vaccine failure occurred.

An outbreak of equine influenza H3N8 in a riding school is described retrospectively with emphasis on diagnosis and putative vaccine failure. In March 1995 an outbreak of equine influenza occurred among 11 horses in a riding school, where most horses had received basic primary immunizations and several booster vaccinations against influenza. Six of the 11 diseased horses had received their last booster vaccination within 5 months of the outbreak. Nevertheless, the influenza infection spread rapidly and clinical manifestations were prominent with frequent, harsh, dry coughing often accompanied by high fever. Nasal swabs were taken from 11 diseased horses. Influenza A virus of the equine H3N8 (equi-2) subtype was isolated from five nasal swab extracts. Stored nasal swab extracts were also retrospectively investigated in two different enzyme immunoassays designed to detect the type-specific conserved nucleoprotein of influenza A viruses, and in a single-tube reverse transcription-PCR (RT-PCR) using a set of primers based on highly conserved regions of the matrix gene of influenza A viruses. Five nasal swab extracts were found positive in a DAS-ELISA and seven in the Directigen((R)) Flu A (DFA) assay, respectively. Two nasal swab extracts from which virus was isolated did not give a positive result in the DAS-ELISA, and one of these also did not give a positive result in the DFA assay. Nine nasal swab extracts were found positive by RT-PCR. Moreover, all virus isolation and/or ELISA positive nasal swab extracts were confirmed by RT-PCR. Three nasal swab extracts were negative by virus isolation, PCR and ELISA. A significant rise in HI titre against influenza A/eq/Miami/63 (H3N8) virus was detected in seven of the nine paired sera available. In acute phase serum samples from 10 horses, SRH antibody levels varied widely. However, some horses with high, or at least putatively clinically protective SRH antibody levels, showed clinical signs and infection was confirmed. Antigenic analysis of two isolates showed that A/eq/Holland/1/95 (H3N8) and A/eq/Holland/2/95 (H3N8) cluster with the UK isolate Osgodsby/92, the Swedish isolate Borlänge/91 and some other European isolates, with H/2/95 identical in reactivity to Borlänge/91 and H/1/95 more similar in reactivity to Osgodsby/92 than H/2/95. Nucleotide and deduced amino-acid sequences showed large differences of both isolates as compared with Miami/63, Fontainebleau/79 and Kentucky/81, the influenza A H3N8 subtype strains incorporated in the vaccines used in this riding school. The role of antigenic drift in vaccine breakdown is discussed in the light of evidence for vaccine breakdown in the UK in 1989, Sweden in 1991 and in the USA since 1991.

Animals↗

[Preliminary results using a combined xylose absorption/hydrogen exhalation test in horses].

In the present study the breath hydrogen (H2) excretion test was combined with the xylose absorption test in 4 normal horses and 9 clinical patients with chronic diarrhea (n = 3) or chronic weight loss without diarrhea (n = 6). All horses underwent a thorough clinical examination. Laboratory evaluations consisted of haematology and serum biochemistry as well as bacteriological and parasitological examination of feces. In addition, serum electrophoresis and abdominocentesis was performed in all the clinical patients. Gastroscopy was carried out in 6 patients and rectal biopsies were obtained from 4 animals. Two animals were euthanized within 4 weeks after the evaluation. The diagnosis of chronic granulomatous enteritis was confirmed in one of them at necropsy, the cause of weight loss in the other remained unexplained. In the remaining animals, specific causes such as management, parasites, chronic infections and diseases of liver, kidneys and heart were excluded, but no specific diagnosis could be obtained. For the combined xylose absorption/H2 excretion test, the animals were starved overnight and given 0.5/kg/bwt xylose as a 10% solution by stomach tube the next morning. Blood samples were collected for plasma xylose analysis at 30 min intervals for 4 hrs, and breath samples were also collected at 30 min intervals for 8 hrs. With the exception of the animal with granulomatous enteritis, the diseased horses showed only slight alterations in either peak concentrations or times to reach peak levels of plasma xylose. None of the healthy animals showed an increase in breath H2 production after xylose administration, whereas five of the diseased animals showed distinct increases of variable heights. In addition, the diseased horses showed higher fasting breath H2 levels (range 7.5-61.5 ppm) than normal horses (range 0-5 ppm). It is concluded that gastrointestinal disorders might be influenced or even induced by a change in intestinal microbial composition, as evidenced by an increased hydrogenic metabolism.

Animals↗

Computerized ECG recording in horses during a standardized exercise test.

In the horse, it is important to make a reliable ECG recording during exercise as cardiac arrhythmia is one of the possible causes of poor performance. The purpose of the present study was to obtain a satisfactory, standardized method for ECG recording by computer during a standardized treadmill exercise test and to evaluate the procedure both in healthy and in diseased horses. Ten experienced healthy research horses were used to develop and adapt the original method (for humans) of computerized ECG recording with the Cardio Perfect software program and to obtain reference values for heart rate and blood variables. Further, three diseased horses (patients) were examined to evaluate whether the test procedures were also usable for horses without extensive experience in treadmill and other experimental procedures. During a standardized exercise test on a treadmill at walk (1.7 m/s), trot (4.0 m/s), and canter (7.0 m/s at 0% and 4% treadmill inclination), the ECG was recorded using two bipolar leads (from xiphoid to manubrium respectively to the left withers) and one unipolar lead (on the left ventral thorax). The quality of the exercise ECGs was satisfactory. The heart rate was detectable and arrhythmias were identifiable. Most research horses did not show arrhythmias during exercise, except a single atrial premature complex (APC). The reference values for the mean heart rate for the different stages of the exercise test differed from 84 + or - 7 to 176 + or - 21 beats per minute (bpm). The exercise test provoked in healthy horses a plasma lactate concentration around the anaerobic threshold. Most patients were also able to finish the test. In conclusion, the developed method to obtain an exercise ECG during a standardized exercise test proved to be reliable and applicable both in research horses and in patients, thus extending the possibilities to diagnose arrhythmias.

Animals↗

Respiratory disease in thoroughbred horses in training: the relationships between disease and viruses, bacteria and environment.

A longitudinal study of respiratory disease in racehorses was carried out to assess its relative associations with different infectious agents and to examine any role that the environmental conditions might play. The relationships between coughing, nasal discharge, pyrexia and lower respiratory tract disease were also examined to provide information for improving clinical diagnosis, particularly of disease of the lower respiratory tract. Lower airway disease was closely associated with infection with Streptococcus zooepidemicus. It was also found that equine herpesvirus seroconversions and S pneumoniae infections were independently associated with the development of nasal discharge. Coughing was a specific, but insensitive measure of lower respiratory tract disease (specificity 84 per cent, sensitivity 38 per cent). However, horses that coughed were very likely to have had lower airway disease for more than one month. Horses housed on straw in loose boxes were twice as likely to suffer from lower airway disease as those kept on shredded paper in American barns. The study was not large enough to assess the significance of rarer infections but it did improve the definition of the problem of respiratory disease in racehorses and revealed some of the trends in the associations between viruses, bacteria and the environment in respiratory disease.

Adenoviridae↗

Amyloid-beta in Alzheimer's disease: the horse or the cart? Pathogenic or protective?

While the pathogenesis of Alzheimer's disease (AD) is unclear, amyloid-beta plaques remain major lesions in the brain of individuals with AD. Likewise, amyloid-beta is one of the best-studied proteins relating to the pathogenesis of AD. Indeed, the pathological diagnosis of AD tends to be congruous with the quantity of amyloid-beta. However, it is important to recognize that pathological diagnosis merely represents the association of a pattern of pathological changes with a clinical phenotype. Therefore, it should be acknowledged that, although amyloid-beta detection and semiquantification have some diagnostic utility, the simple presence of amyloid plaques, as with proteinaceous accumulations in essentially all neurodegenerative diseases, does not presume aetiology. Thus, in this review, we discuss the role of amyloid-beta in the pathogenesis of AD and provide an alternative view to the widely accepted dogma.

Alzheimer Disease↗