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Evaluation of pituitary gland anatomy and histopathologic findings in clinically normal horses and horses and ponies with pituitary pars intermedia adenoma.

OBJECTIVE: To determine size and weight of the pituitary gland and associations between pituitary gland size and weight and sex and age in horses without clinical signs associated with pituitary pars intermedia adenoma (PPIA) and horses and ponies with PPIA. ANIMALS: Pituitary glands from 100 horses without clinical signs of PPIA and 19 horses and 17 ponies with PPIA. PROCEDURES: Pituitary glands were weighed, measured, and examined histologically by use of H&E stain. Masson trichrome and periodic acid-Schiff staining were used, when appropriate. Histologic lesions in the pars intermedia, pars distalis, or both were classified as no significant lesions, single or multiple cysts, focal or multifocal hyperplasia, single or multiple microadenomas, and adenoma. Relative pituitary weight (RPW) was calculated as pituitary weight (grams) divided by body weight (grams). RESULTS: There was an age-related increase in the presence of pituitary lesions in the pars distalis and pars intermedia in geldings, mares overall, and non-pregnant mares. Mean (+/-SD) RPW in horses with PPIA was not significantly different from ponies with PPIA (15+/-5.9 x 10(-6) and 16+/-72 x 10(-6), respectively). Maximum pituitary weight in a horse with PPIA was 13.9 g (RPW, 2.9 X 10(-5)). Plasma glucose concentration was positively correlated with RPW in ponies with PPIA. CONCLUSIONS AND CLINICAL RELEVANCE: Pituitary lesions may be a factor in horses with insulin resistance and laminitis before development of clinical signs of PPIA. Ovarian steroids may be involved in the pathogenesis of lesions in the pars intermedia.

Adenoma↗

Postmortem findings of gastric ulcers in Swedish horses older than age one year: a retrospective study of 3715 horses (1924-1996).

The present study was performed to estimate retrospectively the prevalence of gastric ulceration in necropsied Swedish horses and to evaluate some potential risk factors. Horses (n = 3715) older than age one year and necropsied 1924-1996 were included. Information about breed, gender, age, season of death, clinical signs of colic, Gasterophilus larvae and documentation of diseases in different organs was recorded. Data on size, number and anatomical distribution of gastric ulcers were also obtained. The cold-blooded horses and the Standardbreds were the 2 most dominating breeds in our material. Gastric ulcers/erosions were seen in 633 (17%) of the horses. The highest prevalence was found in Thoroughbreds (19%) and Standardbreds (19%), whereas only 7% of the cold-blooded horses were affected. The cutaneous region along the margo plicatus was the most commonly affected area (52%). Multiple ulcers were seen in 62% of the horses with gastric ulcers and concurrent ulcers in the oesophagus were reported in 6%. In the bivariate analysis, significant associations were found between gastric ulcers and signs of colic (P<0.001) and gender (P<0.001), as well as between concomitant bowel (P<0.001), liver (P<0.05) and oesophageal (P<0.01) lesions. Using a multivariate logistic regression model, only year of necropsy and breed were found significantly to influence the risk of gastric ulceration by decreasing the Log Likelihood test value. The present study shows clearly that gastric ulceration has been present in Swedish horses throughout the 20th century, which also may be the situation worldwide.

Age Factors↗

Palmar digital vessel relaxation in healthy horses and in horses given carbohydrate.

OBJECTIVE: To compare in vitro smooth muscle relaxation of palmar digital vessels from healthy horses with those from horses in the prodromal stage of experimentally (carbohydrate) induced laminitis. ANIMALS: 16 adult horses. PROCEDURE: Segments of palmar digital vessels were obtained from 5 healthy horses and 6 horses given carbohydrate. Vascular rings from the palmar digital artery and vein were suspended in individual organ baths containing buffer solution and indomethacin; isometric tension was recorded, and contraction and relaxation were compared. Smooth muscle contraction in response to cumulative addition of phenylephrine was recorded in the absence and presence of 1 microM NG-nitro-L-arginine methyl ester (L -NAME). After wash out, vascular rings were preconstricted with phenylephrine (0.3 microM), and cumulative endothelium-dependent (acetylcholine-induced) and independent (nitroprusside-induced) smooth muscle relaxations were recorded in the absence or presence of L -NAME. RESULTS: Phenylephrine increased vascular smooth muscle tone in ring preparations of palmar digital arteries and veins. Addition of acetylcholine or nitroprusside induced relaxation of palmar digital artery and vein ring preparations. Use of L-NAME (1 microM) significantly reduced maximal relaxation induced by acetylcholine, but not by nitroprusside. Maximal relaxation induced by acetylcholine, but not by nitroprusside, was reduced in vascular rings prepared from carbohydrate-overloaded horses. CONCLUSION AND CLINICAL RELEVANCE: Reduced endothelium-dependent relaxation of palmar digital vessels may have a role in the pathophysiology of acute laminitis after carbohydrate overload in horses.

Acetylcholine↗

Histamine inhalation challenge in normal horses and in horses with small airway disease.

A histamine inhalation challenge (HIC) procedure was developed to assess hyperreactive states in horses. Following clinical evaluation, percutaneous lung biopsies were performed on nine light breed mares aged 6 to 15 years. Five horses, with normal small airways, were classified as group A and four subjects with small airway disease (SAD) lesions formed group B. Pulmonary mechanics parameters were monitored following an aerosol of 0.9% saline and every 5 min for up to 30 min after HIC with 0.5% w/v of histamine diphosphate, administered through a face mask for 2.5 min. Tidal volume (VT) and airflow (V) values were obtained with a pneumotachograph. Transpulmonary pressure (delta Ppl) was measured by the esophageal balloon catheter method. Dynamic compliance (Cdyn), total pulmonary resistance (RL), end expiratory work of breathing (EEW) and respiratory rate (f) were calculated by a pulmonary mechanics computer. Group A horses had increases in RL, and decreases in Cdyn whereas horses in group B were hyperreactive and showed greater changes in EEW, Cdyn, and delta Ppl but with a relatively lower variation of RL. One horse in clinical remission from SAD, but with a high biopsy score (group B), and one clinically normal horse belonging to group A showed marked hyperreactivity as shown by increases in EEW, maximum change in delta Ppl and RL and decreases in Cdyn. These results suggest that the HIC described can be used as a method to investigate airway hyperreactivity and SAD in horses.

Aerosols↗

Transmission of the South African asinine strain of equine arteritis virus (EAV) among horses and between donkeys and horses.

Lateral and sexual transmission of EAV among horses and lateral transmission between donkeys and horses were attempted by experimental infection with the South African asinine strain. Clinical, immunological and virological responses were evaluated. All intramuscularly inoculated horses developed very mild clinical signs, were viraemic, shed virus from nasopharynx, and seroconverted. Lateral infection was demonstrated in one in-contact mare. Reinfection of two stallions by intranasal instillation was shown by virus recovery from buffy-coat cultures. After nasal instillation of virus, one stallion which did not become infected by in-contact exposure, showed slight serous nasal and ocular discharge, contained virus in a blood and nasopharynx and seroconverted. Attempts to transmit the virus from seropositive stallions to seronegative mares by breeding, were not successful; no virus was isolated from semen. All inoculated donkeys and three in-contact horses showed clinical signs consistent with an EAV infection. Although virus was isolated from donkey buffy-coat preparations and the nasopharynx, and they seroconverted, no virus was isolated from the horses, and they failed to seroconvert; it was assumed that their clinical signs were due to factors unrelated to EAV. The South African strain of EAV appears to be poorly transmissible to horses, supporting the findings of other field studies which indicate a widespread distribution and long-standing presence of the virus among South African donkeys, but a very restricted prevalence of seropositive horses.

Animals↗

Cadmium/zinc relationships in kidney cortex and metallothionein of horse and red deer: histopathological observations on horse kidneys.

Cadmium and zinc were determined in kidney cortex of 63 horses and 51 red deer (Cervus elaphus). Cadmium and zinc were also determined in protein fractions obtained by Sephadex chromatography of kidney cortex from 10 horses and 4 red deer. Histopathological parameters in kidney cortex of horses were compared to cadmium content. The metal contents (on wet weight basis) in kidney cortex of the horses were 0.31 +/- 0.22 mmole Cd/kg (range 0.03-1.21) and 0.63 +/- 0.17 mmole Zn/kg (range 0.36-1.23). The Zn content increased with the Cd content, the Zn increase being less at higher concentrations. No significant increase of the Cd content with age of the horses (range 2-19 years) was found. The metal contents in kidney cortex of the red deer were 0.030 +/- 0.031 mmole Cd/kg (range 0.002-0.13) and 0.51 +/- 0.37 mmole Zn/kg (range 0.30-2.82). Here the Zn content did not increase significantly with Cd content, and in this case a positive linear relation of Cd content with age of the deer (range 0-11 years) was observed. The molar Zn/Cd ratio in the metallothionein fractions of horse kidney cortex was less variable at higher Cd contents. All Cd and the increase of Zn above an average basal level of 0.36 mmole Zn/kg was recovered in the metallothionein fractions. No gross histopathological changes could be observed in kidney cortex of the horses. Nevertheless a slight increase of the thickness of the basement membrane of Bowman's Capsule and also of the diameter of Bowman's Capsule in relation to Cd content up to 0.3 mmole/kg could be observed.

Animals↗

IGF -I plasma concentrations in non-treated horses and horses administered with methionyl equine somatotropin.

Insulin-like growth factor-I (IGF -I) is likely to be an indicator of somatotropin (ST) administration in the horse. To investigate the different ways ST administration may be detected, the following aspects of IGF -I concentrations in plasma were studied: (i) the daily variation; (ii) variation following a treadmill test; (iii) concentrations at rest and after exercise; and (iv) concentrations in plasma from two young horses and two adults treated with methionyl equine somatotropin (e ST). In the population of horses at rest, IGF -I mean concentration (SEM) was 261 (104) ng ml(-1). In post race samples, IGF -I mean concentration was 187 (100) ng ml(-1). All of these data indicate that exercise does not modify IGF -I concentration in plasma. The magnitude of the increase in IGF -I following administration of e ST differed according to the age of the horses. The critical value of 700 ng ml(-1)was exceeded for 1 day in adult horses and for at least 11 days in young horses. These results show that IGF -I has potential as an indirect marker of ST administration in horses.

Age Factors↗

Haematological and biochemical changes in horses competing in a 3 Star horse trial and 3-day-event.

Haematological and biochemical changes in horses competing in the Endurance Test (Phase T and D) of an advanced Horse Trial (HT, n = 22) and the Endurance Test (Phases A-D) of an advanced (CCI) 3-day-event (TD, n = 11) over a similar course on the same day were studied. Environmental conditions during the event were cool (5.5-11.1 degrees C). Blood samples were collected from the horses in each group the evening prior to the Endurance Test, within 60 s after, and 10 min after, completion of Phase D (cross-country jumping). The following were determined in the blood samples and compared between the 2 groups of horses: packed cell volume (PCV), serum total protein [TP], serum albumin [ALB], plasma lactate [lactate], serum total calcium [TCa], plasma ionised calcium [Ca+2], serum inorganic phosphate [PO4], plasma pH, plasma sodium [Na], plasma potassium [K], serum chloride [Cl], serum urea nitrogen [SUN], serum creatinine [Cr] and serum glucose concentrations and aspartate aminotransferase (AST) and creatine kinase (CK) activities. The PCV and [Cr] were higher in the TD group and approached significance (P = 0.063 and P = 0.057, respectively). The [TP], [ALB], [Na], glucose concentration and CK, and AST were significantly higher and [Cl] and [PO4] were significantly lower in the TD group after exercise when compared to the HT group. It was deduced from these data that the horses competing in the 3-day-event experienced greater fluid and electrolyte losses, reduced glomerular filtration, higher glycogenolysis and had greater leakage of enzymes from working muscles during competition than horses competing in the horse trial.

Animals↗

A retrospective case-control study of horse falls in the sport of horse trials and three-day eventing.

REASONS FOR PERFORMING STUDY: Serious injuries to horses and riders in horse trials (HT) and three-day events (3DE) are usually associated with falls of horses, which invariably involve falls of the riders. Many potential causes for these falls have been discussed. OBJECTIVES: The aim of this case-control study was to investigate the risk factors for horse falls on the cross-country phase of horse trials and three-day events. METHODS: Using retrospective data, significant risk factors identified with unvariable analysis (P value <0.2) were entered into a multivariable logistic regression model. Significant risk factors (P value <0.05) were included in the final model. RESULTS: It was revealed that a number of course, obstacle and rider variables were significantly and independently associated with the risk of falling. Falling was associated with obstacles sited downhill (Odds ratio [OR] 8.41) and with obstacles with ditches in front (OR = 5.77). CONCLUSIONS: The relationship between course variables and the risk of falling was characterised and showed a significantly increased risk with increasing numbers of jumps on the course and for jumping efforts later in the course. In contrast, after allowing for the total number of obstacles on the course, an increase in the total number of jumping efforts appeared to have a protective effect. A later cross-country start time was associated with a decreased risk of a horse fall. Amateur event riders were approximately 20 times more likely to fall than professional riders. POTENTIAL CLINICAL RELEVANCE: This study has identified a number of risk factors associated with horse falls and highlights areas that can be altered to improve safety in cross-country competitions.

Accidental Falls↗

[Sequence comparing of mtDNA D-Loop varied region in Chinese Mongolian horse and External Thoroughbred horse].

Mitochondrial DNA D-Loop varied region 400 bp sequence variations in 4 Chinese Mongolian horses and 4 External Thoroughbred horses were analyzed in this experiment. The results showed that the average nucleotide mutational rate of mtDNA D-Loop varied region in 4 Chinese Mongolian horses was 3.69%, while External Thoroughbred horses were 4.00%. Three types of mutations including transition, transversion and deletion were all found in the investigated mtDNA D-Loop regions, of which transition was the most frequent. Nucleotide mutational loci were abundant, length mutations were found and great differences were all observed among the 8 horses. It showed there existed much polymorphism in the mitochondrial DNA D-Loop varied region of Chinese Mongolian horses and External Thoroughbred horses.

Animals↗

Genetic variation of Polish endangered Biłgoraj horses and two common horse breeds in microsatellite loci.

Genetic variation of endangered Biłgoraj horses and two common Polish horse breeds was compared with the use of 12 microsatellite loci (AHT4, AHT5, ASB2, HMS2, HMS3, HMS6, HMS7, HTG4, HTG6, HTG7, HTG10, VHL20). Lower allelic diversity was detected in all investigated populations in comparison to other studies. Large differences in the frequencies of microsatellite alleles between Biłgoraj horses and two other horse breeds were discovered. In all polymorphic loci all investigated breeds were in the Hardy-Weinberg equilibrium. Mean Fis values and the results of a test for the presence of a recent bottleneck were non-significant in all studied populations. Comparable values of observed and expected gene diversity indicate no substantial loss of genetic variation in the Biłgoraj population and two other breeds. The lowest variability observed in the investigated group of Thoroughbred horses was confirmed. About 10% of genetic variation are explained by differences between breeds. Values of pairwise Fst and two measures of genetic distance demonstrated that Biłgoraj horses are distantly related to both common horse breeds.

Animals↗

Detection of equine infectious anemia virus in horse leukocyte cultures derived from horses in various stages of equine infectious anemia viral infection.

The enzyme-linked immunosorbent assay (ELISA) antigen-positive and agar-gel immunodiffusion test (AGID)-negative horses do not have infective equine infectious anemia (EIA) virus. The ELISA testing of horse leukocyte culture (HLC) supernatants did detect EIA virus in a HLC that was infected with the Wyoming strain of EIA virus and in HLC derived from horses in febrile, acute, or subacute stages of EIA infection. In supernatants of HLC derived from chronic and inapparent carrier horses, EIA virus was not detected with ELISA. Direct fluorescent antibody tests detected EIA virus in HLC infected with 10(6)TCID50 of the Wyoming strain of EIA virus and in 50% of the HLC from febrile acute or subacute horses. The direct fluorescent antibody testing of HLC derived from chronic and inapparent carrier horses did not detect cell-associated EIA virus. The pony inoculation test proved to be the most reliable and accurate method for detecting infective EIA virus in horses in various stages of EIA infection and accurately correlated with the AGID test.

Animals↗

[Population genetic parameters of aboriginal Yakut horses as related to modern breeds of the domestic horse Equus caballus L].

This study was the first to analyze the polymorphic characteristics of a wide range of biochemical markers in aboriginal Yakut horses. A total of 124 alleles, including 48 alleles of seven blood-group loci and 76 alleles of ten loci for enzymes and other proteins, were studied. For these polymorphic systems, a computer analysis of the genetic distances between 85 horse breeds of different origin from all parts of the world was performed. The low level of hereditary variation in the Yakut horses confirmed that this breed is old and has long been an isolated population. Phylogenetic analysis demonstrated that Yakut horses exhibited the most genetic similarity to the breeds from the Central Asian cluster, such as Akhal Teke, Arabian, Yabou, and Caspan Pony (Iran). The dispersal route of ancient horses was revealed. It led from America through Siberia and Central Asia to Africa and Eastern Europe, where evidence of the earliest domestication of horses was found. Genetic and ecological explanations of the formation of racing and draft breeds with similar immunogenetic characteristics are advanced. These explanations agree with craniological data on fossils and with the relative rates of growth of the axial and peripheral skeletons in modern breeds. These data shed light on the initial stages of domestication of the horse, an event that was extremely important for development of the human civilization.

Animals↗

Preparation of recombinant African horse sickness virus VP7 antigen via a simple method and validation of a VP7-based indirect ELISA for the detection of group-specific IgG antibodies in horse sera.

This paper describes the production and purification of a group-specific recombinant protein VP7 of African horse sickness virus serotype 3 (AHSV-3) and validation of an I-ELISA for the detection of IgG-antibodies to VP7 in horse sera. Baculovirus-expressed VP7 crystals were purified from infected insect cells. Analytical accuracy of the I-ELISA was examined using sera (n = 38) from an experimentally infected horse, from foals born to vaccinated mares, from guinea-pigs immunized with nine serotypes of AHSV, and from sera of animals infected with other orbiviruses. Compared to traditional serological assays, the I-ELISA was more sensitive in detection of the earliest immunological response in an infected horse and declining levels of maternal immunity in foals. Antibodies to all nine serotypes of AHSV could be detected. Cross-reactivity to related orbiviruses was not observed. Diagnostic accuracy of the I-ELISA was assessed by testing sera from vaccinated horses (n = 358) residing in AHS-enzootic areas and from unvaccinated horses (n = 481) residing in an AHS-free area. Sera were categorised as positive or negative for antibodies to AHSV using virus neutralisation tests. The TG-ROC analysis was used for the selection of the cut-off value. At a cut-off of 11.9 of the high positive control serum (percentage positivity), the I-ELISA specificity was 100%, sensitivity 99.4%, and the Jouden index was 0.99.

African Horse Sickness↗

[Use of the immunoenzyme test ELISA-NS3 to distinguish horses infected by African horsesickness virus from vaccinated horses].

A vaccination protocol involving three horses, with five repeated injections of inactivated serotype 4 African horse sickness virus, was undertaken to determine a possible threshold for the appearance of antibodies against the non-structural protein NS3. Using an indirect enzyme-linked immunosorbent assay, with the recombinant NS3 protein as an antigen, the authors detected a response to NS3 as of the second injection for the first horse and after four injections for the second horse. No response to NS3 was detected for the third horse. The results show that the inactivated vaccine is insufficiently purified to eliminate the non-structural protein NS3. Therefore using the NS3 protein as a marker did not enable differentiation between vaccinated and infected horses.

African Horse Sickness↗

Urinary concentration of corticoids in normal horses and horses with hyperadrenocorticism.

The urinary corticoid:creatinine (c:c) ratio was determined in eight horses with hyperadrenocorticism (HAC). The mean (+/- SD) urinary c:c ratio of the eight horses with HAC (29 +/- 14 x 10(-6)) was significantly (P < 0.05) greater than the ratio in seven control horses (11 +/- 4.3 x 10(-6)). The urinary concentration of corticoids in control horses (201 +/- 60.4 nmol litre-1) was significantly (P < 0.05) lower than in horses with HAC (664 +/- 291 nmol litre-1). The urinary creatinine concentrations did not differ significantly (P > 0.05) between both groups. As both false negative and false positive cases were found, it is concluded that a measurement of the urinary c:c ratio in the horse should not be used as the sole test to confirm equine HAC.

Adrenal Cortex Hormones↗

Pulmonary artery wedge pressure measurement in healthy warmblood horses and in warmblood horses with mitral valve insufficiencies of various degrees during standardised treadmill exercise.

In 12 healthy warmblood horses and 10 horses with mitral valve insufficiencies (MVI) of various degrees heart rate and pulmonary artery wedge pressure (PWP) was measured at rest and during standardised exercise on a high speed treadmill. There was a significant increase in PWP with each change in speed of the treadmill (p < 0.01). The PWP of horses with mild mitral valve regurgitation under working conditions was not significantly different compared to the healthy horses. The horses with moderate mitral valve regurgitation showed a significant higher pulmonary artery wedge pressure at rest and during exercise compared to the healthy horses (p < 0.01) at rest and during treadmill velocity. The tendencies were seen that mild mitral valve regurgitation results only in mild hemodynamic changes during exercise, while moderate MVI have an important influence on haemodynamics.

Animals↗

Effects of platelet activating factor on the distribution of radiolabelled leucocytes and platelets in normal horses and asymptomatic horses with chronic obstructive pulmonary disease.

Antigen challenge is known to cause the recruitment of neutrophils to the lungs of horses with chronic obstructive pulmonary disease (COPD). To evaluate a possible role of platelet activating factor (PAF) in this process, the effects of PAF on the distribution of radiolabelled neutrophils were compared in normal horses and asymptomatic horses with COPD. Changes in lung function, heart rate and the distribution of platelets and eosinophils were also measured. PAF (5 ng kg-1 intravenously) caused immediate but transient increases in the number of radiolabelled neutrophils in the lungs and a concomitant decrease in the peripheral neutrophil count. The total numbers of circulating leucocytes and neutrophils were also significantly decreased by PAF. Rapid and reversible increases in heart rate, respiratory rate and pleural pressure were also observed. In separate experiments, the numbers of radiolabelled eosinophils and platelets in the lungs increased transiently after the administration of PAF. The responses to PAF were qualitatively and quantitatively similar in normal horses and asymptomatic COPD horses. The PAF receptor antagonist WEB2086 (3 mg kg-1 intravenously) inhibited the effects of PAF. These results suggest that PAF, if released in the lungs of horses with COPD during an antigen challenge, might contribute to the recruitment of leucocytes and the respiratory changes.

Analysis of Variance↗