Disturbance of oxidant/antioxidant equilibrium in horses naturally infected with Trypanosoma evansi

Oxidant/antioxidant equilibrium disturbance has already been reported in trypanosome infections by several authors. The present study was aimed to explore the possible oxidant/antioxidant disturbance in surra of naturally infected horses before and after treatment. Fifteen naturally infected horses were chosen to analyse erythrocytic indices, platelet counts, lipid peroxides (LPO), nitric oxide (NO), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) while six healthy animals acted as control. There was a highly significant (P<0.001) reduction in red blood cell (RBC) count, haemoglobin (Hb), packed cell volume (PCV) and platelet levels and a significant reduction in mean corpuscular haemoglobin concentration (MCHC) (P<0.05) was noticed. A highly significant increase in NO (P<0.001), a significant increase in LPO (P<0.05) and a significant decrease in GSH, SOD and CAT (P<0.05) were found. A negative correlation of RBC count with LPO (r=-0.844) and nitrate (r=-0.702) while a positive correlation with GSH (r=0.489), SOD (r=0.580) and CAT (r=0.689) was observed. All the animals were treated with Quinapyramine sulphate (3mg/kg s.c.) only once. Nine animals recovered completely without any side effects. The recovered animals were monitored and samples were collected every seven days for up to 21 days and parameters were analysed. After treatment, a significant increase in haematological parameters was noticed whereas the oxidative indices varied without any statistical significance. To conclude, the increase in oxidant parameters and decrease in antioxidant enzymes in infected horses indicates the disturbance of oxidant/antioxidant indices. There was a significant increase in post therapy haematological values, while the oxidant/antioxidant indices changed insignificantly indicating that antioxidants might be supplemented in the therapeutic regimen.     

Form of Vitamin E Supplementation Affects Oxidative and Inflammatory Response in Exercising Horses

Vitamin E is an essential antioxidant that may benefit athletes by reducing oxidative stress and influencing cytokine expression. Supplements can be derived from natural or manufactured synthetic sources. This study aimed to determine (1) if supplemental vitamin E is beneficial to exercising horses and (2) if there is a benefit of natural versus synthetic vitamin E. After 2 weeks on the control diet (vitamin E–deficient grain and hay), 18 horses were divided into three groups and fed the control diet plus (1) 1000 IU/d synthetic α-tocopherol (SYN-L), (2) 4000 IU/d synthetic α-tocopherol (SYN-H), or (3) 4000 IU/d RRR-α-tocopherol (natural source [NAT]). On day 7, horses began a 6-week training protocol, with standard exercise tests (SETs) performed before and after the 6-week protocol. Venous blood samples were collected on days 0, 7, 29, and 49. Horses fed NAT had higher α-tocopherol (P < .05) at post-SET1 through post-SET2. Plasma thiobarbituric acid–reactive substance levels were lower in NAT versus SYN-L horses after SET2 (P = .02). Serum aspartate aminotransferase was lower after exercise in NAT horses versus SYN-L and SYN-H (P = .02), and less reduction in stride duration was seen after exercise in NAT as compared with SYN-L and SYN-H (P = .02). Gene expression of tumor necrosis factor α was lower in NAT compared with SYN-H (P = .01) but not SYN-L. In conclusion, feeding higher levels of natural vitamin E source resulted in higher serum α-tocopherol levels as well as some improvement in oxidative and inflammatory response and improved functional outcomes in response to an exercise test.     

Prevalence of ulcers of the squamous gastric mucosa in standardbred horses

This study was performed to determine the prevalence of ulcers in the gastric squamous mucosa in Standardbred racehorses. Observations were performed at monthly intervals between the beginning of their training season and their 1st qualifying race. This study also identified risk factors at different levels of race training. Forty-eight Standardbred racehorses from 3 training stables in Quebec, Canada, were studied. Baseline historical information and gastroscopic findings were recorded at the beginning of the trial, and once a month thereafter, between December 2001 and June 2002, until the horse's 1st qualifying race or the end of the training. Intensity of training ranged from jogging to intensive training just before the 1st race and was assigned an ordinal score. Location of squamous ulcers and their appearance were observed on gastroscopy, and an ordinal score was assigned. Prevalence of squamous ulcers from the 2nd through the 4th month (72-88%) of training remained at a significantly higher level (P = .002 to .04) than at the onset of the study (38%) and was also higher in intensely trained horses than in joggers (93% versus 56%). Moderate or more intensive training increased the odds (odds ratio [OR], 3.39; confidence interval [CI], 1.34-8.56; and OR, 11.4; CI, 3.21-40.5, respectively) of detecting ulcers with higher scores. These odds were also higher in trotters (OR, 2.17; CI, 1.07-4.43) than in pacers and generally increased with the duration of training. Duration of training, training level, and gait type also influenced the number of sites with ulcers in the same way. Ulcers had higher scores along the lesser curvature (LC) and the margo plicatus (MP) areas of the stomach. It was concluded that squamous ulcers appeared early in the training of Standardbred racehorses, that the number of sites affected and the ulcer score are related to the intensity of training, and that trotters are more prone to squamous ulcers than pacers.     

Atrial natriuretic peptide and cardiac troponin I concentrations in healthy Warmblood horses and in Warmblood horses with mitral regurgitation at rest and after exercise

ObjectiveAtrial natriuretic peptide (ANP) and cardiac troponin I (cTnI) serve as biomarkers for increased cardiac pressure/volume loading and for myocardial stress or damage. The objective was to describe the time course of plasma ANP concentrations (Cp_ANP) and plasma cTnI concentrations (Cp_cTnI) in horses with mitral regurgitation (MR) compared to healthy horses at rest and after exercise, and to describe the relationship of Cp_ANP with cardiac dimensions and intracardiac pressures.Animals15 healthy Warmblood horses and 7 Warmblood horses with MR.MethodsCardiac dimensions at rest were measured using echocardiography. All horses underwent standardized treadmill exercise. Biomarker concentrations and intracardiac pressures were measured at rest and after exercise. Hypotheses were tested using statistical methods. The level of significance was P < 0.05.ResultsHorses with MR showed increased left atrial (LA) and left ventricular (LV) dimensions but similar exercise capacity compared to healthy horses. Pulmonary capillary wedge pressures (PCWP) and Cp_ANP increased with exercise. Horses with MR had higher PCWP and higher Cp_ANP at rest and after exercise compared to healthy horses, with the maximum difference in Cp_ANP reached 10 min after exercise. Cp_ANP was significantly related to PCWP and – although inconsistently and only in healthy horses – to echocardiographic indices of LA and LV size and function. Cp_cTnI was low throughout the study in both groups.ConclusionsCp_ANP is increased in horses with MR and is related to LA pressures and to left heart dimensions. MR is not necessarily associated with exercise intolerance and exercise-induced myocardial stress or damage.     

Oxidative Stress and Antioxidant Status in Standardbreds: Effect of Age and Training in Resting Plasma and Muscle

The hypothesis of this study was that yearlings would have more oxidative stress as measured by malondialdehyde (MDA), total glutathione (GSH-T), glutathione peroxidase (GPx), and nitric oxide (NO) than mature mares before training, but after they would have similar levels. Ten Standardbred yearling fillies and 10 mature Standardbred mares were split into trained and nontrained groups. Horses were trained 5 days/wk for 7 weeks. Blood and muscle samples were collected at rest on weeks 0, 2, 5, 7, and 9. At week 0, the yearlings had higher muscle GSH-T and NO concentrations (P = .03) than the trained mares. At week 9, plasma NO concentrations were lower in trained mares than in the trained yearlings (P = .007). Trained mares increased muscle MDA and decreased plasma MDA concentrations from weeks 0 to 9 (P < .01), while all mares increased muscle NO and GSH-T concentrations by week 9 (P < .05). Trained mares and yearlings had increased erythrocyte GPx activity at weeks 7 and 9 and GSH-T concentration at week 7 (P < .05). Mares had higher lipid peroxidation and lower antioxidant status in the muscle than the yearlings prior to training. Trained mares improved antioxidant status and oxidative stress levels through training, resulting in levels similar to the yearlings.     

Evaluation of total oxidant and antioxidant status in dogs under different CO2 pneumoperitoneum conditions

Background

The induction of the pneumoperitoneum increases intraabdominal pressure (IAP), causing splanchnic ischemia, whereas its deflation normalizes IAP and splanchnic blood flow. We investigated the oxidant-antioxidant status of dogs who underwent low pressure (7 mm Hg), standard pressure (12 mm Hg), and high pressure (15 mm Hg) pneumoperitoneum.

Results

Twenty-four beagle dogs (12 males and 12 females), 4–6 years old, weighing 8–11 kg were used. The animals were assigned to one of four groups (n = 6 dogs). Group 1 served as a control; these animals received only anaesthesia for 90 min. In groups 2, 3 and 4, intra-abdominal pressure was increased to 7, 12 and 15 mmHg, respectively, and maintained for 60 min. Total oxidant status (TOS) and total antioxidant status (TAS) were determined in venous blood samples. The percentage ratio of TOS to TAS provided an oxidative stress index (OSI).No significant difference in TOS levels was found among the groups. A significant decrease in TAS levels and an increase in OSI levels were observed at 90 min and 24 h of pneumoperitoneum deflation within group 4. No differences were found among the groups.

Conclusions

A high pressure pneumoperitoneum induced significant changes in TAS and OSI. In addition, TOS and TAS levels are useful markers for evaluating changes in the oxidative status caused by a pneumoperitoneum during laparoscopy. Furthermore, a low-pressure pneumoperitoneum could attenuate oxidative stress induced by CO₂ insufflation in dogs.     

Oxidative Stress Biomarkers and Erythrocytes Hemolysis in Well-Trained Equine Athletes Before and After Exercise

The chronic exposure to regular exercise training seems to increase resistance to oxidative stress and improves the antioxidant defense system. The purpose of the present study was to investigate the effect of an exercise test of moderate intensity on oxidative stress biomarkers, antioxidant enzymes activity, and osmotic resistance of erythrocytes in well-trained equine athletes. Eighteen middle-aged horses of Ukrainian warmblood (8.3 ± 1.6 years) and Holsteiner (7.4 ± 1.9 years) breeds were used in this study. All horses have been in regular training for several years. The exercise test induced a significant increase of erythrocyte values, hemoglobin concentration, and hematocrit in horses of both breeds. Regular training induces activation the antioxidant enzymes and thereby can reduce oxidative stress in athletic horses. Our results suggest that the exercise test in horses of both breeds attenuates oxidative stress and accompanied with a significant decrease of lipid peroxidation and oxidatively modified proteins in erythrocytes after exercise. The findings of the present study demonstrated the elevated level of erythrocytes' catalase and glutathione reductase in Ukrainian warmblood horses, as well as decreased level of superoxide dismutase and glutathione reductase in Holsteiner horses reporting changes in levels of exercise-induced oxidative stress biomarkers in horses of both breeds. Statistically significant differences in the percentage of hemolyzed erythrocytes between pre-exercise and postexercise tests were observed and thereby signifying an oxidative stress-dependent impairment of erythrocyte stability. Our data suggest that oxidative stress and enzymatic antioxidant defense biomarkers can be used for the monitoring of fitness level, health benefits, and performance of equine athletes.     

Validation of a method for collection and assay of pentane in the exhaled breath of the horse

Oxidative stress refers to an imbalance between the production of oxidising free radicals and the antioxidant defenses of the cell, and is associated with many pathogenic processes. Oxidative damage to cellular lipids results in the evolution of pentane and ethane gas, and detection of these hydrocarbons in the exhaled breath can be used to monitor in vivo oxidative stress. The aim of this study was to validate a gas chromatography (GC) method for measurement of breath pentane in the horse. The GC-system developed showed good specificity for discrimination of pentane from other breath hydrocarbons, and was sensitive to 0.5 ppb pentane. Pentane was detected in the exhaled breath of five horses investigated on two occasions. The results of this preliminary study demonstrate that breath pentane can be measured in the horse, and further work is now justified to investigate the feasibility of applying this method for monitoring in vivo oxidative stress in the horse.     

Cytokine dysregulation in aged horses and horses with pituitary pars intermedia dysfunction

BACKGROUND: Equine pituitary pars intermedia dysfunction (PPID) is the result of a loss of dopaminergic inhibition of the pars intermedia secondary to neurodegeneration of periventricular hypothalamic neurons. The pathologic events contributing to development of neurodegeneration or clinical signs in equids with PPID are unknown. Chronic inflammation may contribute to initiation or progression of PPID. HYPOTHESIS: Horses with PPID have a distinct systemic cytokine profile compared with that of normal adult or aged horses. The cytokine profile of healthy aged horses differs from that of adult horses. ANIMALS: Aged horses with PPID, healthy aged-matched controls, and adult controls (n = 14 per group). METHODS: Total leukocyte cytokine expression was determined by quantitative polymerase chain reaction (PCR), and tumor necrosis factor (TNF)-alpha plasma concentration was determined by enzyme-linked immunosorbent assay (ELISA). Peripheral blood mononuclear cell (PBMC) TNF-alpha response after endotoxin (lipopolysaccharide [LPS]) treatment was assessed by ELISA. RESULTS: Aged healthy horses had increased expression of interleukin (IL)-6, IL-8, and interferon-gamma as well as PBMC TNF-alpha release after LPS stimulation compared with healthy adult horses. In contrast, aged horses with PPID had increased IL-8 expression, but expression of other cytokines was similar to that of healthy adult horses, not age-matched controls. CONCLUSIONS AND CLINICAL IMPORTANCE: Aged horses show evidence of a proinflammatory state that may contribute to development of age-associated diseases. Horses with PPID have increased expression of IL-8, which may influence the ability of horses with PPID to respond to bacterial pathogens. The general decrease in proinflammatory cytokine expression observed in horses with PPID may be the outcome of high plasma concentrations of anti-inflammatory hormones.     

Plasma homocysteine concentrations in healthy horses and horses with atrial fibrillation

Introduction

Homocysteine (HCY) is an amino acid produced from methionine metabolism. Plasma homocysteine concentrations ([HCY]_p) are elevated (>13 μmol/L) in people with atrial fibrillation (AF) and can predict the recurrence of AF after cardioversion. This study aimed to validate a commercially available human HCY assay for use in horses to develop reference intervals for [HCY]_p and compare [HCY]_p in healthy horses and horses with AF.

Fragmentation of intravenous catheters in three horses

Fragmentation of an intravenous, polytetrafluoroethylene catheter occurred in three horses. The retained fragment was identified in the cervical region by radiography and in the jugular vein by ultrasonography. The retained fragment was removed successfully by jugular venotomy.     

The prevalence of endocrinopathic laminitis among horses presented for laminitis at a first-opinion/referral equine hospital

Endocrinopathic causes of laminitis may be a common underlying causative pathogenesis in first-opinion or field cases presenting with laminitis, as opposed to laminitis produced in inflammatory research models. This study aimed to determine whether evidence of an underlying endocrinopathy was present in horses presented for laminitis to a first-opinion/referral veterinary teaching hospital. A second aim was to compare the signalment of horses and ponies with laminitis with the equine hospital population during the same period. All horses presenting for laminitis at Helsinki University Equine Teaching Hospital, Finland, over a 16-month period were examined for an underlying endocrinopathy. Horses presenting for laminitis were compared with the hospitalized population over the same period. There were 36 horses presented for laminitis, and evidence of endocrinopathy was present in 89%. Of the horses showing an underlying endocrinopathy, one-third had a diagnosis of pituitary pars intermedia dysfunction, and two-thirds showed basal hyperinsulinemia indicative of insulin resistance, without evidence of hirsutism. Phenotypic indicators of obesity were present in 95% of horses with basal hyperinsulinemia without hirsutism. Compared with the hospital population during the same period, horses with laminitis associated with an underlying endocrinopathy were significantly older and more likely to be pony breeds. Our data support that endocrine testing should be performed on all cases of laminitis that do not have a clear inflammatory or gastrointestinal origin.     

Effect of atropine-dobutamine stress test on left ventricular echocardiographic parameters in untrained warmblood horses

The aim of this study was to investigate the effect of combined atropine low-dose dobutamine stress test on left ventricular parameters in adult warmblood horses, to establish a potential protocol for pharmacological stress echocardiography. Seven healthy untrained warmblood horses aged 9 to 22 years were used. Heart rate (HR) and left ventricular B- and M-mode dimensions were recorded at baseline and during stress testing with 35 microg/kg atropine IV followed by incremental dobutamine infusion of 2 to 6 microg/kg/min. HR increased significantly (P < .05) during the pharmacological challenge, and a maximal HR of 156.6 +/- 12.5 bpm was reached at maximal dobutamine infusion rate. Systolic and diastolic interventricular septum thickness, systolic and diastolic left ventricular free wall thickness, and fractional shortening increased significantly and reached a maximum at the highest infusion rate (mean +/- SD: 4.51 +/- 0.27 versus 5.65 +/- 0.31 cm, 2.89 +/- 0.19 versus 3.78 +/- 0.10 cm, 3.72 +/- 0.34 versus 4.77 +/- 0.18 cm, 2.44 +/- 0.28 versus 3.11 +/- 0.34 cm, 34.98 +/- 3.82 versus 50.56 +/- 3.42%, respectively). Systolic and diastolic left ventricular internal diameter decreased significantly during dobutamine infusion. Left ventricular external and internal area were significantly lower at a dobutamine infusion rate of 2 microg/kg/min but no further decrease was observed during the subsequent steps. Systolic and diastolic myocardial area was significantly lower after the administration of dobutamine but not significantly different during dobutamine infusion, when compared to baseline values. This pharmacological stress test induced significant changes in left ventricular echocardiographic parameters in adult warmblood horses. Additional research should evaluate the value of this stress test in horses suffering from cardiac disease.     

Sarcocystis neurona-specific immunoglobulin G in the serum and cerebrospinal fluid of horses administered S neurona vaccine

A vaccine against Sarcocystis neurona, which induces equine protozoal myeloencephalitis (EPM), has received conditional licensure in the United States. A major concern is whether the immunoglobulin G (IgG) response elicited by the vaccine will compromise the use of Western blotting (WB) as a diagnostic tool in vaccinated horses with neurologic disease. Our goals were to determine if vaccination (1) causes seroconversion: (2) causes at least a transient increase in S neurona-specific IgG in the cerebrospinal fluid (CSF); and (3) induces an IgG response that can be differentiated from that induced by natural exposure. Horses included in the study (n = 29) were older than 6 months with no evidence of neurologic disease. The presence or absence of anti-S neurona antibodies in the serum of each horse was determined by WB analysis. Seropositive horses had CSF collected and submitted for cytology, CSF index, and WB analysis. The vaccine was administered to all the horses and boostered 3-4 weeks later. On day 14 after the 2nd administration, serum and CSF were collected and analyzed. Eighty-nine percent (8 of 9) of the initial seronegative horses seroconverted after vaccination, of which 57% (4 of 7) had anti-S neurona IgG in their CSE Eighty percent (16 of 20) of the seropositive horses had an increase in serum S neurona IgG after vaccination. Of the 6 of 20 horses that were initially seropositive/CSF negative, 2 were borderline positive for anti-S neurona IgG in the CSF, 2 tested positive, and 2 were excluded because the CSF sample had been contaminated by blood. There were no WB banding patterns that distinguished samples from horses that seroconverted due to vaccination versus natural exposure. Caution must be used in interpreting WB analysis from neurologic horses that have been recently vaccinated for EPM.     

The effects of feeding and fasting on gastrointestinal sounds in adult horses

The effect of changes in feed intake on auscultatable gastrointestinal sounds has not been systematically studied. Disagreement also is present in the literature about variation in sounds according to the quadrant of auscultation. Gastrointestinal sounds were recorded over the center of the left dorsal, left ventral, right ventral, and right dorsal quadrants and over the middle of the right abdominal flank. During 24 hours (n = 4) or 48 hours (n = 5) of fasting, there was a reduction in the intensity of gastrointestinal sounds as assessed by analysis of sound recordings. There was also a reduction in the number of mixing-like and propulsive-like sounds heard by 2 blinded observers. After refeeding, there was a marked increase in sound. Sound intensity varied among abdominal quadrants, but blinded observers did not notice significant differences in the number of mixing-like sounds. The left dorsal quadrant was quieter than others during fasting and refeeding. The right ventral quadrant appeared to be least affected by fasting, and sounds were louder over the right ventral and right middle quadrants than over the others. The blinded observers' perceptions of sound correlated poorly with one another and with objective measures of sound intensity. This experiment demonstrates the effectiveness of computerized analysis of abdominal sound in detecting a reduction in the intensity of gastrointestinal sounds during fasting and their return during refeeding. The left dorsal quadrant was quieter than other quadrants, likely because of its position over the small colon. There was considerable observer variation in the number of intestinal sounds heard.     

The relationship between serum biotin and oxidant/antioxidant activities in bovine lameness

Serum biotin concentrations, erythrocyte superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and plasma thiobarbituric acid reactive substances (TBARS) were measured in 36 dairy cows, 18 of them were healthy and served as control. In the 18 cows with lameness problems, there were 5 cows with interdigital necrobacillosis, 5 cows with subsolar abscessation, 2 cows with solar ulcers, 2 cows with white line disease, 2 cows with chronic laminitis and 2 cows with septic arthritis. The degree of lameness was estimated to be slight in 3 cows, moderate in 11 cows and severe in 4 cows. Plasma fibrinogen levels and TBARS concentrations were increased significantly (P≤0.05) in lame cows compared to control group. The antioxidant enzymes GSH-Px, and CAT concentrations were increased significantly (P≤0.05) in lame cows. The level of reduced glutathione and the activity of SOD were significantly decreased in affected cows compared to healthy ones. Serum biotin levels in healthy cows ranged from 2.25 to 3.5ng/ml while in lame cows, biotin levels ranged from 1.17 to 2.3ng/ml. Biotin levels correlated positively with blood GSH (r=0.870, P≤0.05), (r=0.735, P≤0.05) and with GSH-Px (r=0.539, P≤0.05), (r=0.637, P≤0.05) and with SOD (r=0.637, P≤0.05), (r=0.449, P≤0.05) and with catalase (r=0.533, P≤0.05), (r=0.585, P≤0.05) in both healthy and lameness affected subjects, respectively.     

Hypomagnesemia in hospitalized horses

This study was initiated to identify the signalment and clinical variables potentially associated with hypomagnesemia in horses evaluated at the North Carolina State University, College of Veterinary Medicine (NCSU-CVM) veterinary teaching hospital between January 1999 and May 2001. A nested case reference study (nested case-control study) was conducted to examine the potential relationship between hypomagnesemia and signalment, serum chemistry panel analyses, number of hospitalization days, discharge status, and diagnosis. A series of independent and multivariable logistic regression models were used to assess the potential association of each variable with low total serum magnesium concentrations. Four hundred one of 823 (48.7%) horses had serum total magnesium concentrations below the normal reference range. Hypomagnesemia was more likely to occur in horses older than I month of age. Colic (odds ratio [OR]: 2.96, 95% confidence intervals [CI]: 2.14-4.08), acute diarrhea (OR: 5.91, 95% CI: 2.32-15.06), other gastrointestinal disease (OR: 2.07, 95% CI: 1.15-3.71), infectious respiratory disease (OR: 5.07, 95% CI: 2.09-12.28), and multiorgan system disease (OR: 2.31, 95% CI: 1.24-4.28) were associated with hypomagnesemia in adult horses, whereas foals with diarrhea (excluding septic foals) (OR: 0.11, 95% CI: 0.01-0.84) were less likely to have hypomagnesemia. Overall, there was no relationship between hypomagnesemia and mortality (OR: 1.00, 95% CI: 0.72-1.41), but horses with colic and hypomagnesemia were less likely to die than horses with colic and normal or high total magnesium (OR: 0.53, 95% CI: 0.30-0.95). Among horses that survived, hypomagnesemia at admission was associated with a longer hospitalization period (OR: 1.45, 95% CI: 1.00-2.11).