Acute phase protein concentrations after limited distance and long distance endurance rides in horses

Acute phase proteins (APP) have been described as useful for assessing health in human and animal patients, as they closely reflect the acute phase reaction (APR). In humans and dogs a reaction analogous to APR has also been described after prolonged or strenuous exercise. The aim of this study was to determine, if similar reactions occur in endurance horses after limited and long distance rides. Seventeen horses that successfully completed various distance competitions were tested. Routine haematological and biochemical tests were performed and the concentrations of serum amyloid A (SAA), C-reactive protein (CRP) and haptoglobin were measured. Typical endurance exercise-induced haematological and biochemical changes were observed in all horses, regardless the distance. After long distance rides, the level of SAA markedly increased, but CRP and haptoglobin concentrations remained unchanged. After limited distance rides no changes in the levels of APPs were noted. Exercise-induced APR in horses occurred only after prolonged, strenuous exertion, and differed from APR in inflammation in that only SAA concentration was increased.     

Racing Induces Changes in the Blood Concentration of Serum Amyloid A in Thoroughbred Racehorses

Intensive exercise results in the increased blood concentration of the acute phase proteins in horses competing in some sport disciplines. In this study, the blood level of serum amyloid A (SAA) was analyzed in Thoroughbred racehorses during 5 days after completion of the race. Samples were collected from 25 healthy Thoroughbred horses beginning with 4 hours after the race and repeated daily up to the fifth day after the race. Serum amyloid A analysis was performed using commercial enzyme-linked immunosorbent assay kit, and the results were presented as median, interquartile range (IQR), and range. Data were analyzed using Friedman's nonparametric analysis of variance. The acute phase response (APR) was reflected by an increased SAA level after the race, reaching significantly higher concentrations on days 1 (P < .001) and 2 (P = .005) and falling below the level of the first sample on day 5 (P = .006). The median peak concentration observed on day 1 after the race was 3.84 mg/L (IQR, 2.32 to 8.86). Racing induces minute changes in SAA concentration typical for the exercise-induced APR; however, the significance of this reaction in the context of horse health and fitness remains unclear.     

Exercised-induced pulmonary hemorrhage in polo and racing horses

Philippine polo and racing horses were examined for exercise-induced pulmonary hemorrhage after their competitive exercise. Exercise-induced pulmonary hemorrhage occurred in 11.1% of the polo horses and 64% of the racing horses. None of the horses had blood at the nostrils.     

Changes in blood constituents accompanying exercise in polo horses

There have been several studies of biochemical changes in horses doing intense exercise such as Thoroughbred and Standardbred racehorses and in horses performing exercise over a long period of time such as endurance horses and three-day eventing horses, but we are not aware of studies with polo horses. Blood samples were taken from 18 polo horses at rest, immediately after playing 2 chukkers of indoor polo, and after a 15 minute rest period. Each horse was studied at 2 different games. The blood samples were analyzed for lactic acid, protein, sodium, potassium, chloride, calcium, phosphorus, HCO-3, PCO2, hemoglobin, packed cell volume, and pH. Samples taken immediately after playing polo had significant increases in lactic acid, protein, sodium, hemoglobin, packed cell volume, and pH, and significant decreases in chloride, calcium, PCO2, and HCO-3. Pulse and respiration were significantly increased. After a 15 minute rest period, there was a significant decrease in potassium. The HCO-3 was lower immediately after playing, but was above the resting value after 15 minutes. It was concluded that the changes after exercise are similar in some aspects to those reported for horses performing intense exercise such as racehorses, and in some aspects to those reported for horses performing prolonged exercise such as three-day event horses and endurance horses. Horses playing indoor polo develop a high plasma lactic acid, but with alkalemia, and could be used as a model to study this condition.     

Aminophylline Affects Glycemia Control and Increases Anaerobic Glycolysis in Horses during Incremental Exercise

A study was conducted on the effects of acute administration of aminophylline on physiological variables in purebred Arabian horses submitted to incremental exercise test. Twelve horses were submitted to two physical tests separated by a 10-day interval in a crossover study. These horses were divided into two groups: control (C, n = 12) and aminophylline (AM, n = 12). The drug at 10 mg/kg body weight or saline was given intravenously, 30 minutes before the incremental exercise test. The treadmill exercise test consisted of an initial warmup followed by gradually increasing physical exigency. Blood samples were assayed for lactic acid, glucose, and insulin. Maximal lactic acidemia was greater (P = .0238) in the AM group. Both V₂ and V₄ (velocities at which lactate concentrations were 2 and 4 mmol/L, respectively) were reduced in the AM group by 15.85% (P = .0402) and 17.76% (P = .0109), respectively. At rest as well as at 4 minutes, insulinemia was greater in the AM group (P = .0417 and .0393). Glycemia was statistically lower in the AM group at times 8 (P = .0138) and 10 minutes (P = .0432). Use of aminophylline in horses during incremental exercise does not seem to be beneficial, because this drug has a tendency to cause hypoglycemia and to increase dependence on anaerobic glucose metabolism.     

Effects induced by exercise on lymphocyte β-adrenergic receptors and plasma catecholamine levels in performance horses

The effect of dynamic exercise on complete blood cell count, lymphocyte β-adrenergic receptor and plasma catecholamine (adrenaline and noradrenaline) levels in horses performing different disciplines were investigated during rest and after exercise. Blood samples were collected from jumping horses (n=6), Arabian Endurance horses (n=6) and Standardbred trotters (n=6) before and immediately after competition. Dynamic exercise caused a significant increase in red blood cell count (Standardbred trotters: P=0.0012), haemoglobin concentration (jumping horses: P=0.001; Standardbred trotters: P=0.01), haematocrit percentage (Standardbred trotters: P=0.005), neutrophil percentage (jumping horses: P=0.0003), lymphocyte percentage (jumping horses: P=0.0003), monocyte percentage (Standardbred trotters: P=0.0008), lymphocyte β-AR numbers (jumping horses: P=0.01; Arabian Endurance horses: P=0.016; Standardbred trotters: P=0.05), plasma adrenaline concentration (Standardbred trotters: P=0.0001) and plasma noradrenaline levels (Standardbred trotters: P=0.003). It is concluded that acute increases in plasma catecholamine concentrations depended on the exercise performed and may induce up-regulation of β-AR in equine lymphocytes. However, the exact mechanism of β-AR up-regulation still remains unclear.     

Acute exercise does not induce an acute phase response (APR) in Standardbred trotters

The purpose of the study was to investigate whether acute strenuous exercise (1600- to 2500-m race) would elicit an acute phase response (APR) in Standardbred trotters. Blood levels of several inflammatory markers [serum amyloid A (SAA), haptoglobin, fibrinogen, white blood cell count (WBC), and iron], muscle enzymes [creatinine kinase (CK) and aspartate transaminase (AST)], and hemoglobin were assessed in 58 Standardbred trotters before and after racing. Hemoglobin levels increased and iron levels decreased 12 to 14 h after racing and haptoglobin concentrations, white blood cell counts, and iron levels were decreased 2 and/or 7 d after racing. Concentrations of CK, AST, SAA, and fibrinogen were unaltered in response to racing.Acute strenuous exercise did not elicit an acute phase reaction. The observed acute increase in hemoglobin levels and decreases in haptoglobin and iron levels may have been caused by exercise-induced hemolysis, which indicates that horses might experience a condition similar to athlete’s anemia in humans. The pathogenesis and clinical implications of the hematological and blood-biochemical changes elicited by acute exercise in Standardbred trotters in the present study warrant further investigation.     

Effect of chronic clenbuterol administration and exercise training on immune function in horses

Effects of longitudinal exercise training and acute intensive exercise (simulated race test) on immune function have not been reported in horses. Clenbuterol, a beta2-adrenergic agonist, is used to manage inflammatory airway disease in horses. This study investigated the interaction of 8 wk of exercise training with or without 12 wk of clenbuterol administration in horses. Twenty-three untrained standardbred mares (10 +/- 3 yr, Mean +/- SE) were used and divided into four experimental groups. Horses given clenbuterol plus exercise (CLENEX; n = 6) and clenbuterol alone (CLEN; n = 6) received 2.4 microg/kg BW of clenbuterol twice daily (in an average volume of 20 mL) on a schedule of 5 d on and 2 d off for 12 wk. The CLENEX group was also aerobically trained 3 d/wk. Mares given exercise alone (EX; n = 5) were aerobically trained for 3 d/wk, and the control group (CON; n = 6) remained sedentary. Both EX and CON horses were administered similar volumes (approximately 20 mL) of molasses twice daily. A simulated race test (SRT) resulted in an elevation in lymphocyte number postexercise (P < 0.05). There was no significant difference after acute exercise in either monocyte or granulocyte number. Acute exercise resulted in a decrease (P < 0.05) in the percentage of CD4+ and an increase (P < 0.05) in the percentage of CD8+ cells. The SRT resulted in a decreased lymphoproliferative response to pokeweed mitogen (P < 0.05). A SRT had no effect on antibody production in response to equine influenza vaccine. The EX group demonstrated greater cortisol concentrations at rest and at all other time points postexercise after completing the training regimen compared with CLENEX horses (P < 0.05). Preexercise (SRT) peripheral blood monocyte number was lower in CLENEX horses than in other treatment groups (P < 0.05). Clenbuterol and exercise training did not significantly affect post-SRT changes in leukocyte numbers. Exercise training resulted in a decrease (P < 0.05) in the percentage of CD8+ cells post-SRT compared with other groups, but the percentage of CD4+ cells was not altered by either clenbuterol or exercise conditioning. Lymphocyte proliferative response was not affected by clenbuterol or exercise treatment. Horses demonstrated responses to bouts of acute exercise as noted with other species, namely humans and rodents.     

Transdermal fentanyl combined with nonsteroidal anti-inflammatory drugs for analgesia in horses

This study investigated the pharmcokinetics, efficacy, and safety of the fentanyl transdermal therapeutic system (TTS) in horses in which there was an inadequate analgesic response to nonsteroidal anti-inflammatory drugs (NSAIDs) alone. Nine horses with pain that was refractory to therapeutic doses of phenylbutazone (n = 3) or flunixin meglumine (n = 6) subsequently also received between 39 and 110 microg/kg of transdermal fentanyl. Blood samples were collected at 0, 1, 2, 3, 4, 5, 6, 12, 24, 36, 48, 60, and 72 hours after patch application, and a radioimmunoassay was used to determine serum fentanyl concentrations. Pharmacokinetic values were determined by noncompartmental analysis. Physical examination findings were recorded in all horses, and pain and lameness grading systems were used to assign scores to 8 and 6 horses, respectively. All horses tolerated the administration of fentanyl TTS, in that no clinically significant adverse effects attributable to fentanyl were observed. Use of the TTS resulted in variable serum concentrations of fentanyl, with a peak serum concentration of 2.2+/-1.1 ng/mL (mean+/-SD) and a time to peak serum concentration of 26+/-13 hours. After transdermal fentanyl administration, mean time to reach serum fentanyl concentrations consistent with analgesia in other species (1 ng/mL) was 14 hours. In addition, serum fentanyl concentrations of 1 ng/mL or greater were maintained in all but one horse for at least 18 hours. Pain scores were significantly decreased after fentanyl TTS and NSAID administration (P < .05), but lameness scores were not significantly different (P > .05). Overall, administration of fentanyl TTS had a favorable pharmacokinetic profile in horses with clinical pain, and the fentanyl TTS in combination with NSAIDs appeared to provide safe and effective analgesia in most of the horses with pain that was refractory to NSAID therapy alone.     

Vaccination elicits a prominent acute phase response in horses

European and American guidelines for vaccination against tetanus and influenza in horses recommend annual and annual/semi-annual vaccinations, respectively, against the two pathogens. Too-frequent vaccination may, however, have adverse effects, among other things because an inflammatory response is elicited with subsequent alterations in homeostasis. The objective of the study was to compare the acute phase response (APR) in 10 horses following administration of two different types of vaccines, namely, an inactivated Immune Stimulating COMplex (ISCOM) vaccine and a live recombinant vector vaccine. Blood was sampled before and after vaccination to measure levels of serum amyloid A (SAA), fibrinogen, white blood cell counts (WBC) and iron. Vaccination induced a prominent APR with increased WBC, elevated blood levels of SAA and fibrinogen, and decreased serum iron concentrations. The ISCOM vaccine caused significantly (P<0.05) greater SAA, fibrinogen and WBC responses than the vector vaccine. During the APR muscle catabolism and liver and kidney metabolism are altered. Also drug metabolism may change during the APR. The findings of the present study may be relevant for advising horse owners about convalescence after vaccination.     

Changes in blood biomarkers in Arabian horses with Clostridium difficile-induced enterocolitis

Clostridium difficile (CD) is considered a major health care problem both in developing and developed countries; frequently reported to be associated with enterocolitis and diarrhea in horses and other animals. In this study, we examined acute phase response (APR), cytokines response, neopterin (NP) procalcitonin (PCT) production and oxidative stress condition in horses and foals with C. difficile-induced enterocolitis (CDIE) and evaluated the effectiveness of these parameters as biomarkers for the disease. A total of 407 Arabian horses in 35 stables were examined between January 2017 to December 2018. Only 24 out of 407 horses showed two or more signs of CDIE. The blood level of serum amyloid A (SAA), haptoglobin (HP), proinflammatory cytokines (TNF-α, IL-6 and IL1-β), serum malondialdehyde (MDA), PCT and NPT in horses with CDIE were higher than in healthy horses. Nevertheless, the levels of nitric oxide (NO), superoxide dismutase (SOD) and total antioxidant concentration (TAC) were considerably lower in diseased horses compared to those that were healthy. The ROC curves for eleven selected blood parameters, both in healthy horses and horses with CDIE demonstrated that all examined blood markers had significant levels of differentiation between CDIE cases and healthy controls (AUC > 87.5). The data in this study suggest that the evaluation of acute-phase proteins, cytokines, PCT, NPT, and oxidative stress biomarkers may well be used as a tool for diagnosis and assessment of CDIE and in disease pathogenesis in Arabian horses.     

Relationship Between Neutrophil Activity,Oxidative Stress,Acute Phase Response,and Lameness Grade in Naturally Occurring Acute and Chronic Joint Disorders in Horses

We hypothesized that in horses with naturally occurring joint disorders, the neutrophil response, acute phase response (APR), and oxidative stress parameters elevated significantly and are markers of increased inflammatory response in these conditions. Therefore, the first aim of the study was to evaluate neutrophil response, oxidative status, and APR. The neutrophil activity was assessed on the basis of elastase, myeloperoxidase (MPO), and alkaline phosphatase release, whereas free radical generation was assessed on the basis of nitric oxide and superoxide production. Acute phase response was estimated on the basis of fibrinogen or haptoglobin plasma concentration and oxidative stress on the basis of malondialdehyde plasma concentration. Then, these parameters were compared with lameness grade, and correlation coefficients between them were calculated. The study was conducted on 43 horses divided on control group of healthy horses (n = 17), acute lameness (AL) group (11 horses), and chronic lameness (CL) group (15 horses). The neutrophil activity from horses of both groups of joint disorders (AL and CL) was significantly (P < .01) higher in comparison with healthy horses. Elastase release was 67.28 ± 1.89% of maximal activity in AL group in comparison with 51.72 ± 1.75% in healthy horses and 62.61 ± 1.54% in CL group. The highest values of other enzymes were also noted in AL group. Moreover, in AL group release of elastase and MPO positively correlated (P < .01) with grades of lameness. These findings revealed the mutual relation between studied parameters and obtained results may be useful in the development of new therapeutic strategies in the treatment of acute and chronic joint disorders in horses.     

The effect of dietary fish oil supplementation on exercising horses

Ten horses of Thoroughbred or Standardbred breeding were used to study the effects of dietary fish oil supplementation on the metabolic response to a high-intensity incremental exercise test. Horses were assigned to either a fish oil (n = 6) or corn oil (n = 4) treatment. The fish oil (Omega Protein, Hammond, LA) contained 10.6% eicosapentaenoic acid and 8% docosahexaenoic acid. Each horse received timothy hay and a textured concentrate at a rate necessary to meet its energy needs. The supplemental oil was top-dressed on the concentrate daily at a rate of 324 mg/kg BW. Horses received their assigned diet for 63 d, during which time they were exercised 5 d/wk in a round pen or on a treadmill. During wk 1, horses exercised for 10 min at a trot. After wk 1, exercise time and intensity were increased so that at wk 5, exercise time in the round pen increased to 30 min (10 min of cantering and 20 min of trotting) per day. Starting at wk 6, horses were exercised 3 d/wk in the round pen for 30 min and 2 d/wk on a treadmill for 20 min. After 63 d, all horses performed an exercise test consisting of a 5-min warm-up at 1.9 m/s, 0% grade, followed by a step test on a 10% grade at incremental speeds of 2 to 8 m/s. Blood samples were taken throughout exercise. During exercise, horses receiving fish oil had a lower heart rate (treatment x time interaction; P < 0.05) and tended to have lower packed cell volume (treatment effect; P = 0.087). Plasma lactate concentrations were not affected by treatment. Plasma glucose concentrations were not different between groups during exercise but were lower (treatment x time interaction; P < 0.01) for the fish oil group during recovery. Serum insulin tended to be lower in fish oil horses throughout exercise (treatment effect; P = 0.064). There was a tendency for glucose:insulin ratios to be higher for fish oil-treated horses throughout exercise (treatment effect; P = 0.065). Plasma FFA were lower (treatment x time interaction; P < 0.01) in horses receiving fish oil than in horses receiving corn oil during the initial stages of the exercise test. Serum glycerol concentrations also were lower in fish oil-treated horses (P < 0.05). Serum cholesterol concentrations were lower in horses receiving fish oil (treatment effect; P < 0.05), but serum triglycerides were not affected by treatment (P = 0.55). These data suggest that addition of fish oil to the diet alters exercise metabolism in conditioned horses.     

Echocardiographic analysis of segmental left ventricular wall motion at rest and after exercise in horses with and without heart disease

An echocardiography was performed on 23 healthy warmblood horses and on 12 warmblood horses with cardiac diseases at rest and after treadmill or lungeing exercise. The B-mode technique was used, and the left ventricular wall motion was analyzed visually. The left ventricle was divided into 6 equally sized myocardial segments, 3 of them in the region of the interventricular septum, and the other 3 in the region of the left ventricular rear wall (“6-segment model”). The kinetic of each segment was determined using a kinetic score system. A normokinetic wall motion was defined with a kinetic index of 1. To the authors' knowledge, the current study is the first to present a visual qualitative method for assessment of the healthy equine heart with regard to the myocardial left ventricular wall motion at rest and after exercise under standardized conditions. First, reference values for the left ventricular myocardial kinetics were established in healthy horses. Subsequently, horses with cardiac diseases of differing severity were examined, and the diagnostic value of the method was evaluated. In contrast with human cardiology, the healthy equine heart is characterized by several deviating wall motion patterns. The majority of the healthy horses (52%) showed a normokinetic wall motion of all 6 segments at rest and a normokinetic concentrically increased hypertrophy after exercise. In 9 healthy horses, however, the middle septal segment (at the papillary muscle level) was characterized by a hypokinesia (kinetics index = 2) at rest. In 5 of these horses, the hypokinesia was not observed after exercise. The kinetic wall motion score was 1.1 ± 0.2 at rest in the healthy horses. In the horses with cardiac disorders, the score was 1.5 ± 0.4 at rest. In all healthy horses, the middle septal segment (at the papillary muscle level) was characterized by a hypokinesia (kinetics index = 2; n = 5) or akinesia (kinetics index = 3; n = 3) at rest.After standardized treadmill exercise, the wall motion score of the healthy horses was 1.12 ± 0.16 and the score of the horses with cardiac disorders was 1.52 ± 0.4. In contrast to human cardiology, the mid-septal segment showed a hypokinesia (kinetics index: 2) and an akinesia (kinetics index: 3) in two healthy horses after exercise. In all horses with low-grade and moderate cardiac disorder, an atypical wall motion pattern was observed after exercise. In all horses with high-grade cardiac disorder, an atypical motion pattern was documented in 5 out of 6 segments. An exercise test was not performed on the latter group.     

Maximal lactate concentrations in horses after exercise of different duration and intensity

Lactate kinetics in whole blood of horses was investigated after exercise of differing velocities and duration. The following categories of exercise were used: A: <11 m/second and >180 seconds (n=35), B: >11 m/second and <180 seconds (n=17) and C: <11 m/second and <180 s (n=10). The mean peak lactate concentration determined in horses in category A was 4.49 ± 2.21 mmol/1, in B, 16.32 ± 4.81 mmoVl and in C, 4.58 ± 1.59 mmol/l. While the maximum lactate concentrations in categories A and C were always found immediately after the exercise, the peaks in category B were measured between the first and tenth minute after exercise. Mean lactate concentrations measured at 2-minute intervals after bouts of category-B exercise tended to stabilize 3 to 10 minutes after exercise; however, mean lactate concentrations measured during the intervals before and after the peak value differed significantly. The lactate concentration returned to pre-exercise levels within 20 minutes after exercise bouts of category C, but remained above pre-exercise levels up to 60 minutes after bouts of category-A and -B exercise. It was concluded that, for an evaluation of lactate data after intensive anaerobic exercise, sequential blood sampling at 2-minute intervals for a period of up to 12 minutes after exercise is necessary. Less frequent sampling may be a reason for the often described irreproducibility of lactate concentrations in horses. After aerobic or mild anaerobic exercise, one sample is sufficient, but it has to be taken as soon as possible after exercise.     

High intensity exercise conditioning increases accumulated oxygen deficit of horses

High intensity exercise is associated with production of energy by both aerobic and anaerobic metabolism. Conditioning by repeated exercise increases the maximal rate of aerobic metabolism, aerobic capacity, of horses, but whether the maximal amount of energy provided by anaerobic metabolism, anaerobic capacity, can be increased by conditioning of horses is unknown. We, therefore, examined the effects of 10 weeks of regular (4-5 days/week) high intensity (92+/-3 % VO2max) exercise on accumulated oxygen deficit of 8 Standardbred horses that had been confined to box stalls for 12 weeks. Exercise conditioning resulted in increases of 17% in VO2max (P<0.001), 11% in the speed at which VO2max was achieved (P = 0.019) and 9% in the speed at 115% of VO2max (P = 0.003). During a high speed exercise test at 115% VO2max, sprint duration was 25% longer (P = 0.047), oxygen demand was 36% greater (P<0.001), oxygen consumption was 38% greater (P<0.001) and accumulated oxygen deficit was 27% higher (P = 0.040) than values before conditioning. VLa4 was 33% higher (P<0.05) after conditioning. There was no effect of conditioning on blood lactate concentration at the speed producing VO2max or at the end of the high speed exercise test. The rate of increase in muscle lactate concentration was greater (P = 0.006) in horses before conditioning. Muscle glycogen concentrations before exercise were 17% higher (P<0.05) after conditioning. Exercise resulted in nearly identical (P = 0.938) reductions in muscle glycogen concentrations before and after conditioning. There was no detectable effect of conditioning on muscle buffering capacity. These results are consistent with a conditioning-induced increase in both aerobic and anaerobic capacity of horses demonstrating that anaerobic capacity of horses can be increased by an appropriate conditioning programme that includes regular, high intensity exercise. Furthermore, increases in anaerobic capacity are not reflected in blood lactate concentrations measured during intense, exhaustive exercise or during recovery from such exercise.     

Evaluation of the acute phase response in cloned pigs following a lipopolysaccharide challenge

The objective of this study was to evaluate the acute phase response (APR) in cloned pigs derived from two different cell lines [C1 (n = 2) and C2 (n = 7)] as compared to genetically similar non-cloned pigs (CONT; n = 11) following a lipopolysaccharide (LPS; 25 microg/kg BW) challenge. Pigs were weaned at 21 days of age and maintained in individual pens in the same room until sample collection approximately 1 week later. Blood samples were collected every 30 min for 2 h prior to and 4h after the LPS challenge. Serum samples were analyzed for cortisol, tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6). Average gestational length for cloned pigs, 118.8 +/- 0.97 days, was longer (P < 0.005) than that of CONT pigs, 114+/-0.41 days. For serum cortisol, there was a time by group interaction (P < 0.0001) such that the cortisol response was greater in CONT pigs as compared to C2 pigs (P < 0.0001), but not different from C1 pigs (P > 0.74). A time by group interaction (P < 0.0001) was observed for serum TNF-alpha such that the TNF-alpha response was greater in CONT pigs as compared to C2 pigs (P = 0.0002) and tended to be greater (P < 0.06) than C1 pigs. A time by group interaction (P < 0.0001) was also observed for serum IL-6 such that the serum IL-6 response was greater (P < 0.003) in CONT pigs as compared to C2 pigs and there was a trend (P = 0.10) for serum IL-6 to be greater in CONT pigs compared to the C1 pigs. These are the first results to demonstrate that cortisol and proinflammatory cytokine profiles associated with the APR of cloned pigs are altered compared to genetically similar non-cloned pigs. Our results also indicate that the cell line from which clones are derived may dictate the APR. The hormone and cytokine profiles reported herein are a significant contribution towards our understanding, and perhaps our ability to prevent or reduce the incidence of premature deaths in cloned animals and warrants further investigation of the immune system of cloned animals.     

Phenylbutazone prevents the endotoxin-induced delay in gastric emptying in horses.

The effect of phenylbutazone on gastric emptying in horses was determined by measuring serum concentrations of acetaminophen. Gastric emptying was determined in normal fasted horses (n = 6), horses given endotoxin intravenously (n = 6), horses given intravenous phenylbutazone (n = 6), and horses given intravenous phenylbutazone plus endotoxin (n = 6). The mean time to reach maximum serum acetaminophen concentration (Tmax), the maximum serum concentration (Cmax), and the area under the serum acetaminophen concentration versus time curve (AUC) were compared among treatment groups. Phenylbutazone did not alter gastric emptying in normal horses. Endotoxin caused a profound delay in gastric emptying, and pretreatment with phenylbutazone abolished this effect.     

Effects of competition on acute phase proteins and lymphocyte subpopulations – oxidative stress markers in eventing horses

The aim of the study was to evaluate markers of the acute phase response (APR) in eventing horses by measuring acute phase proteins (APP) (haptoglobin, Hp, and serum amyloid A, SAA), lysozyme, protein adducts such as pentosidine‐like adducts (PENT), malondialdehyde adducts (MDA), hydroxynonenal adducts (HNE) and total advanced glycation/glycoxidation end products (AGEs), complete blood count and lymphocyte subpopulations (CD4+, CD8+ and CD21+) both at rest and at the end of an eventing competition. Blood samples were collected from eight Warmblood horses (medium age 10 ± 3) during an official national 2‐day event competition at rest (R) and 10 min after the arrival of the cross‐country test on the second day. Exercise caused a significant increase in red blood cell number, haemoglobin, packed cell volume, neutrophils, white blood cell and lymphocyte number; however, these values remained within the normal range. The CD4+ and CD8+ cells significantly increased, whereas the CD21+ lymphocytes decreased; a significant increase in serum SAA, lysozyme and protein carbonyl derivates was also observed. Two‐day event causes significant changes in APR markers such as lysozyme, protein carbonyl derivates (HNE, AGEs, PENT) and lymphocyte subpopulations. The data support the hypothesis that 2‐day event may alter significantly APR markers. Limitations of the study were the relatively small sample size and sampling time conditioned by the official regulations of the event. Therefore, further studies are needed to investigate the time required for recovery to basal values in order to define the possible effects on the immune function of the athlete horse.     

Flow cytometric analysis of blood lymphocyte phenotypes in horses infected with the equine infectious anemia virus

Lymphocyte phenotypes were evaluated in bloodsamples taken from horses in the persistent phase EIA virus infection (n=10), from diseased controls (n=5) and from normal controls (n=10). A single animal in the acute phase of EIA was also studied. Cells were identified using flow cytometry after labelling with polyclonal antibodies to horses immunoglobulins for B-lymphocytes, or monoclonal antibodies (MAbs) to CD4, CD5, CD8 and MHC Class-II antigens. In horses persistently infected with EIA virus, the percentage of CD4+T-lymphocytes is systematically reduced and the percentage of CD8+T-lymphocytes is irregular, ranging from normal to severely reduced. Most of them have low values for cells expressing class-II antigens, but high B-cell percentages. Total CD5+ cell percentages are low in all diseased horses examined compared to normal controls. The acutely-infected foal differed from the persistently infected animals in having an elevated percentage of CD8+ T-lymphocytes, and a severely reduced percentage of B-cells.BSA, bovine serum albumin; EIA, equine infectiousanemia; FITC, fluorescein isothiocyanate; MAb, monoclonal antibody; PBL, peripheral blood lymphocytes; PBMC, peripheral blood mononuclear cells, PBS, phosphate buffered saline.