Evaluation of Factors Contributing to Daily Within-Horse Variation of Plasma Alpha-Tocopherol Concentration

The current study was conducted to evaluate daily within-horse variation of plasma alpha-tocopherol concentration (TOC) (Experiment 1 [EXPT1]) and to determine the effect of a single meal on plasma TOC (Experiment 2 [EXPT2]). In EXPT1, venous jugular blood samples were collected from four pregnant (6–7 months of gestation), light horse mares (8.8 ± 2.9 years of age, mean ± SD; range, 5–11 years of age) between 7:30 AM and 8:30 AM, followed by eight hourly samples, and analyzed for plasma TOC. All horses were fed 5 kg dry matter (DM) grass hay 18 hours before sample collection and received no additional feed during the sampling period. Mean within-horse coefficient of variation for TOC was 3.5% ± 0.01%. In EXPT2, seven mature light horses, four mares and three geldings (4.9 ± 4.4 years of age, mean ± SD; range, 2–14 years of age), were randomly assigned to one of two treatments (fasted [FST; n = 7] or fed [FD; n = 7]), followed by reversal of treatments 24 hours later. At 7:30 AM, FD horses were fed 2.1 kg DM grain-mix concentrate (187 IU vitamin E/kg DM) plus 3.3 kg DM mixed-grass hay, whereas FST horses received no feed. Neither group received any feed for 18 hours before 7:30 AM. Venous jugular blood samples were collected at 10:30 AM from all horses and analyzed for plasma TOC. The mean (±SD) within-horse paired-treatment difference was not significant (0.05 ± 0.12 μg/mL). The results suggest that variation in within-horse plasma TOC is relatively small over an 8-hour period in fasting horses and is unaffected 3 hours after a hay grain meal.     

Efficacy of Hyperimmunized Plasma in the Treatment of Horses with Acute Colitis

Colitis in the adult horse is a life-threatening clinical condition that can be caused by any of several enteric pathogens. This study was conducted to determine whether treating horses with plasma obtained from donors that were hyperimmunized against the common equine diarrheal pathogens Clostridium difficile, Clostridium perfringens, and Salmonella sp shortens the duration of diarrhea in acute colitis. To evaluate the efficacy of plasma treatment, 42 horses with acute onset of diarrhea were studied. Horses were enrolled if they were of age >1 year, duration of diarrhea at presentation was <72 hours, and they had not received equine plasma within the last 3 months. In addition, the serum cortisol concentrations of horses with acute diarrhea were studied.Horses were randomized to receive hyperimmunized plasma, control plasma (collected from nonimmunized horses), or no plasma therapy. Clinical parameters and fecal consistency were observed until resolution, discharge, or death, and complete blood counts (CBCs) and biochemical profiles were collected throughout the study. A total of 38 horses completed the study. The mean duration of diarrhea was 40.7 ± 9.8 (mean ± SEM) hours, 119.2 ± 56.1 hours, and 72.0 ± 24.5 hours for the hyperimmunized plasma, normal plasma (NP), and control groups, respectively. Using survival analysis techniques, this difference did not achieve statistical significance (P = .374). Serum cortisol was found to be increased in all horses at presentation and to decrease with time in all treatment groups. There was no difference in cortisol concentrations between the three treatment groups studied (P = .237).     

Salivary Cortisol Concentration in Exercised Thoroughbred Horses

Both physical activity and stress result in an increase in plasma cortisol level. The measurement of cortisol in plasma requires taking blood samples, which is stressful itself. Therefore, the aim of this study was to evaluate the use of saliva sampling for the determination of cortisol concentrations, indicating the intensity of exercise in horses during race training. Twelve Thoroughbred horses aged 2-3 years were examined during their speed training sessions. The horses galloped on the 1,200-m sand track at a speed of 14.4-15.3 m/s. Three saliva samples and three blood samples were collected from each horse. Both types of samples were taken when the horse was at rest, immediately after returning from the track and 30 minutes after the end of exercise. Blood lactic acid (LA) concentration was determined using the enzymatic cuvette test. The concentrations of cortisol in saliva and plasma samples were measured by enzyme immunoassay methods. Statistically significant correlations were found between salivary cortisol level determined 30 minutes after the end of exercise and blood LA concentration obtained immediately after exercise (P = .003) and between salivary and plasma cortisol levels measured 30 minutes after the end of training session (P = .015). The measurement of cortisol concentration in saliva samples taken from race horses 30 minutes after the end of exercise can be recommended for use in practice under field conditions to estimate the level of relative intensity of exercise in race horses.     

Effects of feed form on growth and blood glucose in weanling horses

Eight weanling Standardbred horses were used to evaluate the effects on growth and plasma glucose of two differently processed diets with identical ingredients fed at a rate of 3% of body weight, as fed, daily. The daily ration was divided into two equal feedings at 7:00 am and 7:00 pm. The hay and grain diet (diet HG) consisted of 50% alfalfa hay cubes and 50% of a commercial growing horse texturized grain mix fed as is, without any further processing, with each part fed in separate containers. The completely pelleted diet (diet P) consisted of the same proportions of the hay cubes and grain mix that was ground, pelleted, and fed in a single container. Two groups of four horses were fed either diet P or diet HG for two 21-day periods such that each horse consumed each diet for 21 days. On the last day of each feeding period, 7-mL blood samples were drawn into heparinized evacuated blood tubes for determination of plasma glucose concentration. Blood samples were drawn 30 minutes before the afternoon feeding, immediately before, and every 30 minutes thereafter for 5 hours. Analysis of variance found greater (P = .027) average daily gain for horses fed diet P versus diet HG. A nonsignificant difference was seen in feed efficiency (P = .057) for horses fed diet P versus diet HG. Plasma glucose changes were compared across treatments using area under the curve analysis. No difference (P > .05) was found in the postprandial plasma glucose changes between diet treatments.     

Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.     

Effect of Sward Height on Pasture Nonstructural Carbohydrate Concentrations and Blood Glucose/Insulin Profiles in Grazing Horses

Six mature stock-type geldings with maintenance only requirements were used in a randomized cross-over design to determine the effect of sward height on pasture plant nonstructural carbohydrate (NSC) concentrations and blood glucose and insulin concentrations. Horses were randomly assigned to one of two tall fescue (Lolium arundinaceum Schreb cv Max-Q, Pennington Seed, Madison, GA) grazing cells (0.37 ha) having two different sward heights for a period of 7 days: (1) short (approximately 15 cm; n = 3) or tall (between 30 and 40 cm; n = 3). After the first 7-day period, treatment groups were reversed by moving horses to ungrazed cells having similar characteristics to those used in the first 7 days, so that all horses receive all treatments resulting in six observations per treatment. Both short and tall grazing cells were mowed to a height of approximately 15 cm 32 days before the experiment starts. The short grazing cells were removed to approximately 15 cm at 11 days before the start of the first 7-day period and again 1 day before the start of each 7-day period. All horses had access to pasture for 10 h/d beginning at 8 AM and ending at 6 PM. Although not at pasture, all horses were individually housed in 3.7 × 12.2 m partially covered pens containing automatic water troughs and a crushed stone surface. Herbage mass (kg DM/ha) was determined by use of a falling plate meter for each pasture to ensure that both groups of horses had adequate dry matter to provide grazing for at least 7 days. On day 7 of each period, jugular venous blood samples were collected from each horse before being turned out to pasture, and then at 2, 4, 6, and 8 hours after turn-out. Pasture samples were also collected from each grazing cell at the same time blood samples were taken. Serum and plasma from blood samples were harvested and analyzed for insulin and glucose concentrations, respectively. Pasture samples were analyzed for water soluble carbohydrate (WSC), ethanol soluble carbohydrate (ESC), and starch. The sum of WSC and starch were used as an estimate of NSC. Area under the curve (AUC) and peak concentration were calculated for both plasma glucose (PPG) and serum insulin (PSI) concentration and were analyzed using analysis of variance for randomized cross-over designs. Pasture WSC, ESC, starch, and NSC concentrations were analyzed using analysis of variance for randomized complete block design. A P value of < .05 was considered significant. Mean pasture plant NSC, WSC, and ESC concentrations were lower (P < .001) in short as compared with tall. Pasture plant starch concentration was not different between treatments. Mean pregrazing plasma glucose concentrations, PPG concentrations, and plasma glucose AUC were not affected by treatment. Mean pregrazing serum insulin concentrations were not affected by treatment. Mean PSI and insulin AUC were greater (P < .01) when horses grazed tall, as compared with short. In conclusion, decreasing the sward height by mowing pasture decreased NSC, WSC, and ESC concentrations and subsequently decreased the postprandial insulin response of horses grazing the pasture. These findings may be important in developing strategies aimed at preventing insulin resistance in grazing horses.     

High glycemic and insulinemic responses to meals affect plasma growth hormone secretory characteristics in Quarter Horse weanlings

Growth hormone is a key component of the somatotropic axis and is critical for the interplay between nutrition, regulation of metabolic functions, and subsequent processes of growth. The objective of this study was to investigate potential relations between meal feeding concentrates differing in the glycemic responses they elicit and GH secretory patterns in young growing horses. Twelve Quarter Horse weanlings (5.4 ± 0.4 mo of age) were used in a crossover design, consisting of two 21-d periods and two treatments, a high-glycemic (HG) or low-glycemic (LG) concentrate meal, fed twice daily. Horses were individually housed and fed hay ad libitum. On the final day of each period, quarter-hourly blood samples were drawn for 24 h to measure plasma glucose, insulin, non-esterified fatty acids, and GH. Growth hormone secretory characteristics were estimated with deconvolution analysis. After a meal, HG-fed horses exhibited a longer inhibition until the first pulse of GH secretion (P = 0.012). During late night hours (1:00 AM to 6:45 AM), HG horses secreted a greater amount of pulsatile GH than LG horses (P = 0.002). These differences highlight the potential relations between glycemic and insulinemic responses to meals and GH secretion. Dietary energy source and metabolic perturbations associated with feeding HG meals to young, growing horses have the potential to alter GH secretory patterns compared with LG meals. This may potentially affect the developmental pattern of various tissues in the young growing horse.     

Effect of Restricted Grazing on Hindgut pH and Fluid Balance

Six mature idle geldings were used in a crossover design to determine the effects of restricted grazing on hindgut pH and fluid balance. Initially, horses were randomly assigned to a control group (CTRL: n = 3) having access to warm-season grass pasture continuously, or a restricted grazing (RG: n = 3) group having access to pasture for 12 consecutive hours (1900-700 hours) per 24-hour period for 7 days; they were then reassigned to the opposite treatment for an additional 7 days (i.e., CTRL: n = 6; RG: n = 6). Fecal samples were collected from each horse at 700 hours on day 7 of each period and analyzed for pH and dry matter (DM) as indicators of hindgut pH and fluid balance, respectively. Jugular blood samples were also collected at 700, 1300, and 1900 hours on day 7 and were analyzed for plasma protein as an indicator of systematic fluid balance. Fecal pH and DM data were analyzed using a paired t test. Plasma protein data were analyzed as a repeated-measures design. The mean (± SE) difference between CTRL and RG for fecal pH (.01 ± .16) and fecal DM (.68 ± .6%) was not significant (P = .93 and .52, respectively). Mean plasma protein concentrations were not affected by treatment or by treatment × sample time interaction, but tended to increase (P = .07) during the sampling period regardless of treatment. In conclusion, 12 hours of grazing restriction followed by 12 hours of grazing did not negatively impact hindgut pH or fluid balance.     

Heart rate, blood biochemistry and performance of horses competing in a 100 km endurance ride

To determine whether a clinical examination was adequate to assess the fitness of horses in an endurance ride, and to characterise the relationship between a clinical assessment of the horse's fitness and its blood biochemistry, 23 horses were monitored before, during and after an 87 to 100 km endurance ride. Blood samples were taken before the ride, at three checkpoints on the ride, immediately after the ride and one day later for the determination of packed cell volume, total protein, lactate, glucose, non-esterified fatty acids, creatine phosphokinase, aspartate aminotransferase and plasma electrolytes. Heart rate measurements appeared to be a reliable indicator of the metabolic status of endurance horses suggesting that veterinary examinations according to the official Fédération Equestre Internationale rules are adequate to protect the health and welfare of horses competing in endurance races.     

Leptin and Ghrelin and the Indices of Lipid Metabolism as Related to Sex Steroid Hormones in Trotters

This study examined the influence of sex steroid hormones on lipid metabolism in horses. The group of 34 clinically healthy Standardbred trotters aged 2 to 4 years was studied during an exercise test. The horses were divided into groups according to their sex. These groups were: 11 stallions, 16 mares, and seven geldings. Concentrations of testosterone, 17-β-estradiol, leptin, ghrelin, glycerol, free fatty acids (FFA), and triacylglycerols (TG) were measured in plasma obtained from blood samples taken at rest and after the end of the exercise. At rest, plasma ghrelin concentration was significantly higher in geldings than in stallions and mares (1,541 ± 206 vs 1,280 ± 288 and 1,310 ± 267 pg/mL, respectively; P = .012). Leptin was lower in geldings than in mares (2.65 ± 0.93 vs 4.70 ± 2.31 ng/mL; P = .036). The post-exercise rise in plasma ghrelin and TG concentrations was significantly higher in mares than in geldings (+220 ± 330 vs -25 ± 206 pg/mL; P = .049 and 0.31 ± 0.14 vs 0.13 ± 0.15 mmol/L; P = .016, respectively). The increase in plasma FFA level was higher in geldings than in stallions (535 ± 178 vs 334 ± 191 μmol/L, P = .046). In conclusion, lipolysis rate in geldings is higher than in noncastrated trotters.     

Effects of transport, lairage and stunning on the concentrations of some blood constituents in horses destined for slaughter

The aim of the present study was to determine, in horses destined to slaughter, the effects of transport, lairage and stunning on some blood constituents related to stress. Twenty one horses culled from a race track due to poor athletic performance, transported weekly to a nearby slaughterhouse were used. Each horse was canulated through venipuncture of the right jugular vein with a central venous catheter which remained permanently until after the exsanguination. Through the catheter, blood samples were taken at 6 sampling times: one hour before loading of the horses, immediately after loading, at the end of the journey, immediately after unloading, after lairage, in the stunning box before stunning and during exsanguination. Blood samples were analyzed for the following variables: plasma lactate concentration, plasma creatine phosphokinase activity (CK), plasma glucose concentration, plasma cortisol concentration and packed cell volume (PCV). Time in the stunning box, number of stunning attempts, presence of return to consciousness signs and time between stunning and sticking were also recorded. The results for pre-mortem variables were: mean time in the stunning box 9 min 48 sec, 85.7% of the horses fell at the first stunning attempt, 57.2% of the horses showed signs of returning to consciousness after stunning and the most frequent interval between stunning and sticking was 1.01 to 2 min. Mean values for the blood variables showed a significant rise (P < 0.05) in the concentrations of lactate, glucose and cortisol, as well as CK activity and PCV during transport. The highest values for these blood constituents were seen before stunning and during exsanguination, except for cortisol concentration, which showed the highest values during the transport process.     

Assessment of Insulin and Glucose Dynamics by Using an Oral Sugar Test in Horses

Straightforward testing procedures are needed to facilitate the diagnosis of insulin dysregulation in horses because hyperinsulinemia and insulin resistance are associated with laminitis. Results of an oral sugar test (OST) were compared with those of the intravenous glucose tolerance test (IVGTT). We hypothesized that OST and IVGTT area under the curve values for glucose (AUCg) and insulin (AUCi) would be closely correlated, as defined by a correlation coefficient value ≥0.90. Both tests were performed in 10 horses meeting the criteria for equine metabolic syndrome (EMS) and 8 Quarter horse crossbred mares from a university teaching herd (control group). The OST was also performed in 21 Quarter horse crossbred mares from the same herd, and test repeatability was evaluated in 8 of these horses. All testing was performed under fasting conditions. Median AUCg and AUCi values were 1.3- and 9.0-fold higher, respectively, for the IVGTT and 1.3- and 6.8-fold higher, respectively, for the OST in the EMS group than those in the control group. AUCg (Spearman correlation coefficient [r_s] = 0.58; P = .012) and AUCi (r_s = 0.90; P < .001) values for the two tests were positively correlated. Mean ± SD coefficients of variation for repeated tests in 8 mares were 6.4% ± 3.1% and 45.1% ± 36.2% for AUCg and AUCi, respectively. We conclude that OST and IVGTT insulin results are closely correlated, so the OST warrants further consideration as a field test for insulin dysregulation in horses.     

Sex differences in the response of yearling horses to handling by unfamiliar humans

In adult horses, pronounced sex differences in behavior exist, and many riders prefer male horses to mares. In the present study, we tested the hypothesis that sex differences in the response of horses to handling emerge before puberty, that is, in sexually immature animals. Warm-blooded yearling horses (6 males and 9 females) were exposed to a stationary human test (exposure to an unfamiliar person for 5 minutes) and a tolerance test (haltering and direct contact with the horse) on 5 consecutive days. The horses' behavior and heart rate were recorded. Male yearling horses displayed significantly less exploratory behavior than females during the stationary human test on days 1 and 2 (P < 0.05). Latency to halter and ease with which horses could be haltered were significantly affected by day (P < 0.05), and a significant interaction of day and sex was detected (P < 0.05; e.g., time for haltering: females, day 1: 23.8 ± 7.8 seconds [±standard error of the mean] and day 5: 21.1 ± 1.7 seconds; males, day 1: 53.6 ± 10.4 seconds and day 5: 19.4 ± 2.2 seconds). Heart rate of the horses in response to the test situations was significantly affected by day (P < 0.01) but not by sex. In conclusion, sex differences in behavior do not only exist in adult horses but are already present in young horses before puberty. When exposed to unfamiliar humans for the first time, male yearling horses display more caution than females.     

Age- and Sex-Related Modifications of Hematology in Spanish Purebred Horse

The purpose of this study was to test the hypothesis that age and sex significantly modulate the changes in hematological parameters of physiologically healthy Spanish Purebred horse, establishing the reference values. Blood samples were obtained on a total of 159 horses (n = 77 males and n = 82 females), classified in four age groups: group A (1–2 years; n = 39), group B (>2–3 years; n = 38), group C (>3–4 years; n = 41), and group D (>4–7 years; n = 41). By means ADVIA 2020i, red blood cell (RBC), hemoglobin concentration (HB), packed cell volume, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration, platelets (PLTs), as well as total and differential white blood cells (WBCs) were determined. Total plasma protein was determined by spectrophotometry. Results obtained showed that sex had a limited influence on these parameters, being the HB higher in males and WBC higher in females (P = .047). Age caused a reduction in RBC, with a compensatory increase of MCV and MCH, decrease in WBC, PLT, lymphocytes (LYMPs), as well as an increase of N/L ratio (P = .047). Interactions between age and sex showed higher RBC in males than in females of the group B, and higher HB and WBC in males than in females of the group D. According to hematological data, Spanish Purebred horse was classified as a warm- or cold-blooded horse. Decline of RBC, WBC, and PLT parameters is a natural condition of horse with the advance of age, indicating a decrease of the narrow bone response.     

Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

Selected Biochemical Indicators of Equine Rhabdomyolysis in Arabian Horses: Acute Phase Proteins and Trace Elements

Although creatine kinase (CK), aspartate transaminase (AST), cytokines, and oxidative stress parameters were shown to be useful biomarkers for diagnosis of equine rhabdomyolysis, additional biomarkers of the disease may be of interest to indicate prognosis of the disease. Therefore, the present study investigated acute phase proteins and trace elements as additional biomarkers of ER. Sixty male horses (4-6 years old) of 2 equal groups were used. Horses of the first group were clinically healthy and served as a control group, whereas horses of the second group were ER-diseased animals. Harvested sera were used for estimation of the activities of CK, AST, lactate dehydrogenase (LDH), serum amyloid A (SAA), haptoglobin (Hp), ceruloplasmin (Cp), copper, iron, and zinc, whereas plasma samples were used for determination of fibrinogen. The present findings revealed a significant (P ≤ .05) increase in values of CK, AST, and LDH in diseased horses compared to those in control values. In addition, a significant (P ≤ .05) increase in SAA (162.6 ± 5.32 mg/L), Hp (3.6 ± 0.54 g/L), Cp (39.32 ± 2.31 mg/L), and copper (28.36 ± 1.23 μmol/L) along with a significant (P ≤ .05) reduction in levels of iron (9.32 ± 0.23 μmol/L) and zinc (8.65 ± 1.02 μmol/L) was recorded in diseased horses compare to that in controls (11.3 ± 2.2 mg/L, 0.8 ± 0.2 g/L, 24.23 ± 1.32 mg/L, 18.41 ± 1.03 μmol/L, and 14.2 ± 0.42 μmol/L, respectively). In conclusion, SAA, Hp, Cp, copper, and zinc were useful prognostic biomarkers for the diagnosis of ER in Arabian horses.     

The Effect of the Acute-Phase Response on In Vitro Drug Metabolism and Plasma Protein Binding in the Horse

The effect of the acute-phase response (APR) on the activity of the hepatic drug-metabolizing system (DMS) and on the binding of phenylbutazone to plasma proteins was investigated in the horse. An APR was induced by intramuscular injections of Freund's complete adjuvant in five horses and, five days later, these horses together with five clinically normal horses were shot and the right ventral lobe of each liver removed. The hepatic microsomal fractions from the liver samples ere isolated and significantly lower (p<0.01) concentrations of cytochromes P450 and b5 and activities of aniline-p-hydroxylase and aminopyrine N-demethylase (43%, 55%, 45% and 30%, respectively) were measured in the livers from the adjuvant-inflamed horses, compared to the controls. Phenylbutazone (PBZ) was administered intravenously (4.4 mg/kg) to a further four horses and plasma protein binding was measured by ultracentrifugation. Five weeks later, these horses were injected with Freund's complete adjuvant and the intravenous administration of PBZ (4.4 mg/kg) was repeated. Inflammation induced a significant increase (p<0.01) in the unbound fraction of PBZ (5.2±0.5 as against 1.4±0.6%). These results suggest that the APR depresses the hepatic DMS and reduces the binding of PBZ to plasma proteins.     

Detection and pharmacokinetics of tetrahydrogestrinone in horses

The anti-doping rules of national and international sport federations ban any use of tetrahydrogestrinone (THG) in human as well as in horse sports. Initiated by the THG doping scandals in human sports a method for the detection of 3-keto-4,9,11-triene steroids in horse blood and urine was developed. The method comprises the isolation of the analytes by a combination of solid phase and liquid–liquid extraction after hydrolysis and solvolysis of the steroid conjugates. The concentrations of THG in blood and urine samples were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
A THG excretion study on horses was conducted to verify the method capability for the analysis of postadministration urine samples. In addition, blood samples were collected to allow for determination of the pharmacokinetics of THG in horses. Following the administration of a single oral dose of 25 μg THG per kg bodyweight to 10 horses, samples were collected at appropriate intervals. The plasma levels of THG reached maximal concentrations of 1.5–4.8 ng/mL. Twenty-four hours after the administration plasma levels returned to baseline. In urine, THG was detectable for 36 h. Urinary peak concentrations of total THG ranged from 16 to 206 ng/mL. For the 10 horses tested, the mean plasma clearance of THG was 2250 mL/h/kg and the plasma elimination half-life was 1.9 h.     

Relationships between body weight,fasting blood glucose concentration,sex and age in tree shrews (Tupaia belangeri chinensis)

The tree shrew (Tupaia belangeri chinensis) is a squirrel‐like lower primate or a close relative of primates, commonly used as an animal model in biomedical research. Despite more than three decades of usage in research, the clear relationships between body weight, fasting blood glucose concentration, sex and age among tree shrews remain unclear. Based on an investigation of 992 tree shrews (454 males and 538 females) aged between 4 months and 4 years old, we found that male tree shrews have significantly higher body weight and fasting blood glucose concentration than female tree shrews (p < 0.001). The concentration of fasting blood glucose slightly increased with body weight in males (r = 0.152, p < 0.001). Meanwhile, in females, the body weight, concentration of fasting blood glucose and waist circumference positively increased with age (p < 0.001). Additionally, 17 tree shrews with Lee index [body weight (g)*0.33*1000/body length (cm)] above 290 had significantly higher body weight, waist circumference and glycated haemoglobin A1c (HbA1c) than non‐obese tree shrews with a Lee index score below 290 (p < 0.001). Interestingly, 6 of 992 tree shrews (three males and three females, 2–4 years old) displayed impaired plasma triglycerides, HbA1c, low‐density lipoprotein and oral glucose tolerance test, suggestive of the early symptoms of metabolic syndrome. This study provides the first clear relationships between body weight, fasting blood glucose concentration, sex and age in tree shrews, further improving our understanding of this relationship in metabolic syndrome (MetS). Given the similarity of tree shrews to humans and non‐human primates, this finding supports their potential use as an animal model in the research of MetS.     

Gender Differences–Induced Changes in Serum Hematologic and Biochemical Variables in Mangalarga Marchador Horses After a Marcha Gait Competition

The aim of this study was to investigate whether marcha exercise results in changes in serum hematologic and biochemical variables and whether this condition differs in male and female horses. Thirty-five Mangalarga Marchador horses, 18 males and 17 females were included in the study. Blood samples were obtained before and immediately after a marcha competition. Samples were used for measurement of hematocrit (HCT), hemoglobin concentration (HGB), red blood cell (RBC), total and differential white blood cell (WBC), total protein (TP), albumin (Alb), aspartate aminotransferase (AST), gamma glutamyltransferase (GGT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatinine (Crea), and urea (BUN) levels. Data were submitted to analysis of variance using the SAS statistical program, and means were compared by the Tukey test (P < .05). In both males and females, RBC, HCT, AST, and GGT activity increased in response to exercise, but there were no significant differences between genders. In females, significant increases in TP, BUN, Crea, CK, and LDH levels were detected after the marcha gait, but no such changes were detected in males. Total WBC concentrations increased in response to the exercise in both, males and females; however, there were no differences between genders. Plasma HGB, Alb, and ALP levels after marcha gait did not differ in both genders. Marcha exercise–induced significant hematologic and biochemical changes in serum of Mangalarga Marchador horses, independently of gender. However, females presented more blood changes after marcha gait. Gender differences should be taken into consideration in experiments with athletic horses.