Effects of phlebotomy on haemodynamic characteristics during exercise in standardbred trotters with red cell hypervolaemia

Five Standardbred trotters with red cell hypervolaemia (RCHV) were compared before and after removal of approximately 22% (36 ml/kg bwt) of the total blood volume in order to evaluate the haemodynamic responses, haemorheological alterations and oxygen transport during exercise to fatigue. Data were recorded during submaximal exercise at 4 different speeds on a treadmill and then during continued running at the highest speed step until fatigue. Oxygen uptake (VO2), pulmonary artery pressure (PAP), systemic artery pressure (SAP), heart rate (HR), haematocrit and haemoglobin concentrations (Hb) were measured. Arteriovenous O2 content difference (C(a-v)O2), pulmonary vascular resistance (PVR) and total systemic resistance (TSR) were calculated. Whole blood and plasma viscosity and erythrocyte aggregation tendency were determined with a rotational viscometer. Endoscopy was performed after exercise. ANOVA was used for statistical analysis. Phlebotomy resulted in a decrease in haematocrit and Hb during the course of exercise. Blood and plasma viscosity were lower and erythrocyte aggregation tendency was higher after phlebotomy. Throughout exercise, including submaximal work and continued running to fatigue, PAP, SAP, PVR, TSR and C(a-v)O2 were lower after phlebotomy. HR was higher after phlebotomy during submaximal exercise. Oxygen delivery and VO2 were lower after phlebotomy in the period from submaximal exercise to fatigue. Run time to fatigue was shorter after phlebotomy. Four horses showed exercise-induced pulmonary haemorrhage (EIPH) before phlebotomy and the degree of bleeding was diminished but not abolished after phlebotomy. The reductions in PVR, TSR, PAP and SAP after phlebotomy were probably a result of reduced blood viscosity. In conclusion, although a 22% reduction in blood volume improved the haemodynamic and haemorheological parameters and the degree of EIPH, it was found that RCHV trotters have to rely on high oxygen delivery to the working muscles for maintenance of maximal performance.     

Two-dimensional and M-mode echocardiographic measurements of cardiac dimensions in healthy standardbred trotters

The aim of the study was to establish normal echocardiographic values of healthy Standardbred trotters not published previously. Twenty-three clinically normal horses weighing between 350 and 490 kg were examined in the same manner: first a thorough physical and then detailed echocardiographic examination were performed. Standardised two-dimensional (2D) and guided M-mode echocardiographic imaging techniques were used to measure interventricular septal thickness (IVS), left ventricular internal diameter (LVID), left ventricular wall thickness (LVW), left atrial internal diameter (LAID) in end-systole (s) and end-diastole (d) and aortic diameter (AOD) in end-diastole. Mean, range and standard deviation of the different parameters were calculated. The mean values (in centimetres) were as follow (2D/M-mode): IVSs: 4.6/4.7; IVSd: 3.1/3.0; LVIDs: 7.0/7.0; LVIDd: 10.7/10.7; LVWs: 3.9/3.9; LVWd: 2.7/2.7; LAIDs: 10.4/-; LAIDd: 11.3/-; AODd: 7.2/-. Results of two-dimensional and M-mode measurements were compared to each other and to normal values obtained from other breeds.     

The effects of short intensive exercise on plasma free amino acids in standardbred trotters

The aim of this study was to investigate the effect of short intense exercise on plasma amino acid concentrations in trotters and to test the repeatability of plasma amino acids concentration in samples obtained on two independent days under field conditions. Plasma amino acid concentrations were analysed in blood samples of 36 standardbred trotters before and after intense exercise over a distance of 2000 m. Sampling was repeated in 20 horses after 35 days. Exercise intensity was estimated from post‐exercise lactate levels. Horses were divided in two groups according to a cut‐off lactate concentration at 15 mmol/l. The plasma concentrations of alanine, aspartate, glutamate, isoleucine, leucine, lysine and taurine increased and arginine, asparagine, citrulline, glutamine, glycine, histidine, methionine, serine, tryptophan and 3‐methylhistidine decreased after exercise. Ornithine, threonine, tyrosine, phenylalanine and valine concentrations remained constant. Higher intensity of exercise significantly decreased tryptophan and increased taurine concentrations. Sampling day had a significant effect on the absolute pre‐ and post‐exercise amino acid concentrations. Exercise had a significant influence on the concentrations of most plasma amino acids in trotters. These changes could reflect shifts between the free amino acid compartments, but there were also some indications for muscle catabolism. The amino acid supply of sporting horses could be of specific significance for maintaining muscle integrity and for the improvement of post‐exercise recovery of competition horses.     

Correlations between potassium, calcium and magnesium in blood and muscle of standardbred trotters

This study was conducted to investigate the relationships between the concentrations of potassium, sodium, calcium and magnesium of plasma and muscle in horses, as interactions between these ions can be important in the development of muscle cell injury. Blood samples and gluteal muscle biopsies were taken from 20 healthy Standardbreds to determine the potassium concentration in plasma (cK⁺(P)), red blood cells (cK⁺(RBC)), and muscle (cK⁺(M)), and the calcium and magnesium concentrations in plasma and muscle. No correlations were found between cK⁺(P), cK⁺(RBC), and cK⁺(M). Significant positive correlations were found between cK⁺(M) and muscle magnesium (p<0.01); and cK⁺(P) and plasma calcium (p<0.005). Significant negative correlations were found between cK⁺(M) and the percentage of muscle water (p<0.005); and the muscle magnesium and calcium concentrations (p<0.02).     

Production of reactive oxygen species in neutrophils after repeated bouts of exercise in standardbred trotters

Six trained Standardbred trotters exercised on a racetrack on 2 days with a 3-day interval. On both exercise days the horses trotted three different exercise bouts with increasing intensity with 60-min intervals. Exercise-induced stress was manifested as leucocytosis, an increase in the neutrophil:lymphocyte (N:L) ratio, and increased capacity to produce reactive oxygen species in the peripheral blood as indicated by an increase in whole blood chemiluminescence. The leucocytosis was mainly due to neutrophilia, which lasted for 6 h. Production of reactive oxygen species per single neutrophil showed no significant change during a day of exercise, but was lower on the second exercise day. The cortisol concentrations and N:L ratio, used as indicators of stress, behaved differently: Cortisol did not change significantly after exercise, whereas the N:L ratio increased. These results suggest that in trained horses, the N:L ratio is a sensitive indicator of stress of short duration, and an attenuated N:L response can be taken as an indicator of adaptation to exercise stress.     

Lameness in three-year-old standardbred trotters —Influence of parameters determined before three years of age

The aim of the study was to use available information about a group of Standardbred trotters collected during the first three years of life to predict the risk of lameness in these horses when they reach three years of age. In a previous study on the same population, a sire index defined as the frequency of lameness within each progeny group and presence of radiographic changes other than palmar/plantar bony fragments in the metacarpo- and metatarsophalangeal joints, were found to be significant predictors of the likelihood of developing lameness. The additional variables used in the present study were training and feeding of the horses, information that was collected retrospectively up to the time of clinical examination.A total of 265 three-year-old Standardbred trotters were randomly sampled for an epidemiological study of lameness. Eighty horses were classified as sound and 185 horses as lame, with moderate and severe signs in 138 and 47 horses, respectively. The sire index was found to be significant for predicting lameness (P<0.01), the risk of lameness increasing approximately 2.0 times for every 10% increase in sire index. Training and dietary parameters did not influence the risk of lameness significantly.     

Sclerosis of the third carpal bone. A prospective study of its significance in a group of young standardbred trotters

To assess the significance of radiographic signs of sclerosis of the third carpal bone (C3) in young Standardbred trotters in relation to performance, lameness and bone turnover both carpi in 14 Standardbred trotters were radiographically and scintigraphically examined 6 times, from the beginning of speed training until the beginning of racing, between the mean ages of 20 and 42 months. At the end of the study 8 horses had raced in official qualifying races and 14 limbs in 11 horses had been diagnosed with carpal lameness. All horses but 2 developed sclerosis and all but one had increased bone turnover in the C3 area by scintigraphy. C3 sclerosis increased continuously over time and with increased performance. Carpal lameness was significantly associated with progression of sclerosis but in most cases sclerosis developed without concomitant signs of carpal lameness. No association between carpal lameness and increased scintigraphic uptake was found, but horses that had qualified for racing had significantly higher C3 to carpus ratio of radiopharmaceutical uptake. We conclude that there is a continuous increase in C3 radiographic sclerosis with time in young Standardbred trotters in professional training, but radiographic sclerosis appears to be of limited value as an indicator of clinical carpal disease or level of performance in Standardbred trotters.     

Biochemical markers of bone activity in young standardbred horses during different types of exercise and training

Seven untrained Standardbred horses were used in a training programme of 6 weeks to evaluate the effects of exercise and training on bone metabolism. The horses were exercised on a treadmill according to a standardized exercise test (SET 1: six incremental steps, 5 min duration each; start 5 m/s, increase 1 m/s). SET 1 was followed by a training programme of 6 weeks. In alternating order: high-speed exercise (HSE): 15 min duration, start at VLa4, continuous increase in speed every 60 s by 0.3 m/s (14 incremental steps); low-speed exercise (LSE): constant velocity at VLa2.5, duration: approximately 60-90 min (total training programme: eight HSE and eight LSE sessions). SET 2 finished the training programme and a deconditioning period of 12 weeks followed. Blood samples for lactate, total plasma protein (TPP), osteocalcin, and ICTP (cross-linked C-telopeptide of type I collagen) were collected. ICTP increased during SET 1 and SET 2, whereas osteocalcin decreased to below resting concentration 24 h after SET 1. A rise in ICTP was observed during LSE 1 and LSE 8, which was followed by a drop 24 h after exercise. No changes in osteocalcin were noted during LSE 1, but 24 h after LSE 1 osteocalcin dropped to below pre-exercise levels. LSE 8 resulted in an increase in osteocalcin, followed by a drop 24 h after LSE 8. Osteocalcin and ICTP were not affected by HSE. Baseline osteocalcin levels dropped during the course of training. The acute response of biochemical bone markers indicates a direct influence of a single bout of exercise on bone metabolism.     

Adhesion of Streptococcus gallolyticus strains to extracellular matrix proteins

Fourteen pigeon Streptococcus gallolyticus strains of differing virulence, were tested for their ability to adhere to immobilised fibronectin, collagen types I, III and IV. Eight, 2 and 13 strains were able to bind fibronectin, collagen types III and IV, respectively. None of the strains adhered to collagen type I. Heat treatment, proteolytic digestion or periodate treatment reduced the binding of S. gallolyticus to fibronectin and collagen type IV, suggesting that surface receptors contain proteins and carbohydrates. Although binding to these extracellular matrix proteins can play a role in the pathogenesis of streptococcosis in pigeons, binding properties could not be related to virulence, indicating that other factors determine differences in virulence among pigeon S. gallolyticus strains. Adhesion to collagen type IV may account in part for the distribution pattern of the lesions observed in naturally and experimentally infected pigeons.     

Effects of training on biochemical values in standardbred horses.

Effects of training at a regular, fixed, standard exercise load on venous lactic acid, mixed venous and arterial blood gases and pH, and serum muscle enzymes were determined on previously unconditioned, healthy, adult, Standardbred horses. Arterial and mixed venous blood gases, pH, and serum muscle enzymes did not change in a consistent manner during training. Venous lactic acid concentrations did increase significantly with training and may be of value for the biochemical evaluation of fitness in horses.     

Physical response of dogs supplemented with fish oil during a treadmill training programme

The rise in popularity of dog sports competitions has led to the evaluation of improvements in dog physical performance. The potential benefit of dietary supplementation with fish oil (FO) on the physical performance of human beings and horses has been reported. However, such effect has not been studied in dogs. We therefore evaluated the effect of FO dietary supplementation on heart rate (HR), rectal temperature (RT) and thigh circumference (TC) in dogs during aerobic treadmill training, and further determined HR response and blood lactate (BL) concentration during an incremental exercise test. Using a cross‐over design, eight male dogs were randomly assigned to two groups and received a standard balanced commercial diet (control, CG, n = 7) and the same diet supplemented with 54 mg FO/kg metabolic weight per day (FOG, n = 8). All dogs had 30‐min treadmill sessions at 8 km/hr and 7.5% slope twice a week for 12 weeks. Assessment of HR and RT was performed before and immediately after each session; HR was also assessed 5 min after the end of each session. Thigh circumference was evaluated before each session. All dogs performed an incremental exercise test on the treadmill at 0, 6 and 12 weeks to evaluate HR response and BL concentration. Data were analysed using the mixed procedure (SAS 9.4). In FOG, pre‐HR (?4.9%) and post‐HR (?2.4%) values and post‐RT (?0.3%) values were lower during treadmill training, whereas TC (+2.2%) values were higher as compared with CG (p < 0.01). Through the incremental exercise test, mean HR (week 6, ?5.3%; week 12, ?6.0%) values in FOG were lower than in CG (p < 0.05). In conclusion, FO supplementation slightly improved the physiological response of dogs to exercise during training.     

Role of beta1 integrins in adhesion of canine mastocytoma cells to extracellular matrix proteins

The interactions of tumor cells with the extracellular matrix (ECM) are a crucial step in invasion and metastasis. Integrins are adhesive molecules forming heterodimers with alpha and beta subunits that play a definitive role in these interactions. In this study, mastocytoma (mast cell tumor: MCT) cell-ECM interaction was investigated using 3 canine MCT cell lines: CM-MC (originating from cutaneous MCT), VI-MC (originating from intestinal MCT), and CoMS (originating from oral MCT). Flow cytometric analysis showed that all cells highly expressed the integrin beta1 and alpha1 through alpha5 subunits and that they moderately expressed the alpha6 subunit. In adhesion studies, CoMS weakly but spontaneously adhered to fibronectin (FN), which was enhanced by phorbol ester (TPA), while CM-MC and VI-MC required cell activation by TPA to adhere to FN. Anti-beta1 and alpha5 integrin antibodies strongly inhibited cell adhesion to FN in CM-MC and CoMS and moderately inhibited cell adhesion in VI-MC. Only VI-MC adhered to laminin (LN) under activation by TPA. Anti-beta1 integrin antibodies strongly inhibited cell adhesion to LN, but all anti-alpha integrin antibodies failed to inhibit cell adhesion to LN. No cells adhered to collagen types I and IV. Canine MCT cells from different origins expressed similar integrin patterns; however, there were some differences in adhesive behavior in response to various ECM proteins and activating stimuli.     

Structural remodelling of the heart valves extracellular matrix during embryo development

Alterations in heart valve development represent more than 20% of congenital cardiovascular malformations. Most of the functional properties of heart valves depend on extracellular matrix. Despite its relevance, little is known about fibrillar components on developing stages. Our objective is to define histological changes on valves fibrillar components in late embryonic development of Mus musculus. We found type III collagen as the predominant fibre type in the ECM in prenatal stages followed by a switch to a type I predominance for postnatal ages. The change in fibrillar components is necessary to support the normal mechanical function of adult heart valves.     

Insulin-like growth factor binding proteins initiate cell death and extracellular matrix remodeling in the mammary gland

We have demonstrated that insulin-like growth factor binding protein-5 (IGFBP-5) production by mammary epithelial cells increases dramatically during forced involution of the mammary gland in rats, mice and pigs. We proposed that growth hormone (GH) increases the survival factor IGF-I, whilst prolactin (PRL) enhances the effects of GH by decreasing the concentration of IGFBP-5, which would otherwise inhibit the actions of IGFs. To demonstrate a causal relationship between IGFBP-5 and cell death, we created transgenic mice expressing IGFBP-5, specifically, in the mammary gland. DNA content in the mammary glands of transgenic mice was decreased as early as day 10 of pregnancy. Mammary cell number and milk synthesis were both decreased by approximately 50% during the first 10 days of lactation. The concentrations of the pro-apoptotic molecule caspase-3 was increased in transgenic animals whilst the concentrations of two pro-survival molecules Bcl-2 and Bcl-x were both decreased. In order to examine whether IGFBP-5 acts by inhibiting the survival effect of IGF-I, we examined IGF receptor- and Akt-phoshorylation and showed that both were inhibited. These studies also indicated that the effects of IGFBP-5 could be mediated in part by IGF-independent effects involving potential interactions with components of the extracellular matrix involved in tissue remodeling, such as components of the plasminogen system, and the matrix metallo-proteinases (MMPs). Mammary development was normalised in transgenic mice by R3-IGF-I, an analogue of IGF-I which binds weakly to IGFBPs, although milk production was only partially restored. In contrast, treatment with prolactin was able to inhibit early involutionary processes in normal mice but was unable to prevent this in mice over-expressing IGFBP-5, although it was able to inhibit activation of MMPs. Thus, IGFBP-5 can simultaneously inhibit IGF action and activate the plasminogen system thereby coordinating cell death and tissue remodeling processes. The ability to separate these properties, using mutant IGFBPs, is currently under investigation.     

Efficacy of a paste formulation of omeprazole for the treatment of naturally occurring gastric ulcers in training standardbred racehorses in Canada

The efficacy of a paste formulation of the H+, K+, -ATPase inhibitor omeprazole was evaluated in standardbred racehorses for the treatment and prevention of gastric ulcers. Twenty standardbred racehorses in training, aged 2 to 9 years, were enrolled from 2 training centres in this field trial. Endoscopic examinations confirmed the presence of gastric ulcers in all horses, prior to allocation and treatment and on day 0. Lesions were scored on a scale of 0 to 3 (intact epithelium to extensive ulceration). Replicates were formed, based on training level and location. Within replicates, 1 horse was assigned to group 1 and 3 horses were assigned to group 2, randomly. Horses in group 1 were sham-dosed controls. Horses in group 2 were given omeprazole paste orally at 4 mg/kg bodyweight (BW)/day from day 0 to day 27 and 2 mg/kg BW/day of omeprazole paste orally from day 28 to day 57. Follow-up endoscopies were conducted on post treatment days 28 and 58 or 59. Physical examinations, including BWs, were conducted on all horses prior to treatment and on days 13 or 14, 28, 42 or 43, and 58 or 59. Horses treated with omeprazole had significantly (P < 0.01) more improvement in gastric lesion scores than did controls at day 28 and at study termination on days 58 or 59. All of the omeprazole-treated horses were improved relative to baseline ulcer score at both examinations, and 73.3% were healed (lesion score of 0) at both examinations. None of the controls improved at any point during the study. When the dose was reduced to 2 mg/kg BW, 80% of the horses showed no recurrences or worsening in gastric ulcers. It was concluded that omeprazole paste at 4 mg/kg BW orally, once daily is highly effective in healing gastric ulcers in standardbred racehorses in training and that a dose of 2 mg/kg BW orally, once daily, effectively prevents the recurrence of gastric ulcers in most horses.