Correlation of Plasma Insulin Concentration with Laminitis Score in a Field Study of Equine Cushing's Disease and Equine Metabolic Syndrome

This study aimed to investigate endocrinologic test values and the response to treatment of two commonly encountered causes of endocrinopathic laminitis, equine Cushing's disease (ECD) and equine metabolic syndrome (EMS), in a veterinary practice setting. In particular, the study aimed to determine whether insulin concentration correlated to the severity of clinical laminitis in horses with EMS or ECD. Twenty-five horses were included in the study and assigned to one of three groups: ECD (n = 6), EMS (n = 10), and controls (n = 9). Blood samples were collected at an initial visit and then at regular intervals for the next 12 months. Plasma concentrations of adrenocorticotropin (ACTH), cortisol, and insulin and serum concentrations of glucose and total thyroxine (T4) were obtained. Horses with ECD had significantly higher plasma ACTH concentrations than EMS horses or controls. Horses with EMS and ECD both had significantly higher plasma insulin concentrations than control horses, which was correlated with the Obel grade of laminitis (r = 0.63). After treatment, there was a trend for a reduction in plasma ACTH concentration in horses with ECD. A program of diet and exercise for horses with EMS resulted in reductions in both plasma insulin concentrations and bodyweight, which was variable, depending on the individual. There was a significant correlation between the change in plasma insulin concentration and Obel grade of laminitis (r = 0.69). This study has highlighted the importance of baseline plasma insulin concentration as a potential indicator of the susceptibility of horses to laminitis and the response to a program of diet and exercise.     

Seasonal Variation in Serum Concentrations of Selected Metabolic Hormones in Horses

Background: Determination of adrenocorticotropic hormone (ACTH) concentration is a commonly used test in the evaluation of endocrine causes of equine laminitis, but the concentration in healthy horses can be high at certain times of year, which alters the specificity of the ACTH test. Objective: To determine if circulating concentrations of ACTH, cortisol, glucose, insulin, and thyroxine vary month to month in healthy horses and in horses with equine metabolic syndrome (EMS). Animals: Nine healthy adult horses were studied on their farm/stable over the course of 1 year. After the diagnosis of EMS, 10 laminitic horses residing at the same farm/stable were also studied. Methods: Prospective study of healthy and laminitic horses. Plasma/serum samples were analyzed for concentrations of hormones and glucose. Results: ACTH was the only analyte to show a discrete seasonal pattern, with concentrations in healthy and EMS horses frequently outside of the reference range (9–35 pg/mL) in August through October. Insulin was elevated (>40 μIU/mL) in EMS horses during most months and median serum glucose was generally higher in EMS horses (100 mg/dL, range, 76–163 mg/ dL) than in controls (94 mg/dL, range, 56–110 mg/dL), but no seasonal patterns for insulin or glucose were found. Conclusions and Clinical Importance: An increased ACTH concentration in horses in late summer or autumn should be interpreted with caution. In contrast, insulin concentration is maintained within the reference range throughout the year in healthy horses, thus an increased insulin concentration at any time of year should raise suspicions of EMS, ECD, or both.     

Fat Supplementation to Alfalfa Diets for Refeeding the Starved Horse

The objective was to examine the metabolic responses of chronically starved horses using two isoenergetic diets consisting of either 100% alfalfa hay (AH) or 80% alfalfa hay with 20% corn oil (AO), on a caloric basis. Two 10-d trials were conducted using 14 emaciated, but otherwise healthy, horses with an initial mean body condition score of 1.8 ± 0.9 and mean BW of 336 ± 31 kg. The initial DE intake of 50% of maintenance was gradually increased to 100% during the 10-d refeeding period. Two of the horses receiving the AH diet were euthanized, thus 12 horses (AH, n = 7; AO, n = 5) survived the 10-d study. One preprandial and four postprandial venous blood samples were collected daily. Dietary effects (P<0.01) were shown for blood serum or plasma free fatty acids (FFA), phosphate, magnesium, and 2,3-diphosphoglycerate (2,3-DPG), but not glucose, insulin, or potassium concentrations. The response of insulin and FFA during the 10-d study showed differences (P<0.05) between AH and AO diets. Serum phosphate concentration in horses on both diets exhibited a general slow decline with concentrations below normal reference range during d 5 through 10, thus indicating hypophosphatemia. Serum magnesium increased in horses consuming the AH diet, whereas concentrations in horses consuming the AO diet consistently were below reference range, leading to mild hypomagnesemia. Hypokalemia did not develop in either treatment group, as all serum potassium concentrations remained in the normal reference range. The substitution of corn oil in isoenergetic alfalfa diets was not advantageous in minimizing or preventing hypophosphatemia or hypomagnesemia for refeeding starved horses. The AH diet is recommended as more supportive in refeeding starved horses because of the greater intake of dietary phosphorous and magnesium.     

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.     

Glucose and insulin response after intravenous and subcutaneous somatostatin administration in healthy horses

Equine metabolic syndrome (EMS) is prevalent in the equine population, and somatostatin analogs might be useful for diagnosis and/or treatment of EMS in horses. The purpose of this study was to evaluate the glucose and insulin responses to subcutaneous and intravenous administration of somatostatin. Six healthy research horses were included in this prospective study. An initial pilot study was performed to assess several different doses (10–22 µg/kg [4.5–10 µg/lb]) in two horses, then a final dosage of 22 µg/kg (10 µg/lb) was administered to six horses IV and SQ in a two‐period randomized cross‐over study performed over a 3‐month study period. Blood samples were collected for measurement of plasma insulin and glucose concentrations during a 24‐hr study period. Both IV and SQ somatostatin resulted in decreased insulin and increased glucose concentrations. SQ somatostatin resulted in a longer clinical effect, with return to baseline insulin occurring at 1.5 hr postadministration, versus 45 min for IV. Both IV and SQ administration of somatostatin to normal horses resulted in decreased insulin and increased glucose concentrations, likely due to suppression of insulin secretion by somatostatin. A more prolonged effect was seen following SQ administration as compared to IV administration, and no adverse effects were noted at varying doses. This study provides additional information regarding the effect of somatostatin administration on insulin and glucose concentrations in clinically healthy horses.     

Serum insulin concentrations in horses with equine Cushing's syndrome: response to a cortisol inhibitor and prognostic value

REASONS FOR PERFORMING STUDY: Serum insulin concentration and its use as a prognostic indicator in horses with equine Cushing's syndrome (ECS) have been poorly documented. OBJECTIVES: To examine daily insulin variations in horses with ECS and the effect of treatment using trilostane, a competitive inhibitor of 3beta-hydroxysteroid dehydrogenase. Further, we aimed to examine the relationship between baseline serum insulin concentration and survival in horses with ECS. METHODS: Basal serum insulin concentrations were measured in 20 confirmed ECS cases by taking blood at regular 4 h intervals for 24 h (1200, 1600, 2000, 2400, 0400 and 0800 h) before treatment (Day 0) and 10 days, and 30 days and 1-2 years after the onset of trilostane therapy. The temporal pattern of insulin was analysed using a linear mixed model approach, and the prognostic value of measurements on Day 0 assessed using receiver-operating characteristic analysis. RESULTS: Horses with ECS showed a diurnal pattern of serum insulin concentration, highest value at 1200 h, and this pattern was not altered by treatment with trilostane. Furthermore, despite a mild increase of serum insulin concentrations after 10 days of trilostane therapy, insulin concentration was unaffected in the long-term. Low serum insulin concentrations at the beginning of the trial were significantly associated with improved survival to 1-2 years. The 1200 h sampling before treatment had the highest prognostic value for prediction of survival with a sensitivity and specificity of at least 90% for serum insulin at < 62 and > 188 microu/ml to predict survival and nonsurvival, respectively. CONCLUSIONS AND POTENTIAL RELEVANCE: Insulin is a useful prognostic indicator for ECS, but potentially large variations can occur throughout a 24 h period, indicating a single sample may not be representative. Serum insulin concentration did not increase over 1-2 years in horses receiving trilostane therapy.     

Serum concentrations of calcium, phosphorus, magnesium and calciotropic hormones in donkeys

OBJECTIVE: To provide reference values for serum biochemical variables that are used for evaluation of mineral metabolism in donkeys and compare values with those in horses. ANIMALS: 18 donkeys and 18 horses. PROCEDURES: Total calcium (tCa), total magnesium (tMg), and inorganic phosphorus (P) concentrations were measured in serum samples via spectrophotometry. Ionized calcium (iCa) and magnesium (iMg) concentrations were quantified with selective electrodes. By use of a micropartition system, tCa and tMg were fractionated to separate protein-bound (pCa, pMg) and ultrafiltrable fractions. Complexed calcium (cCa) and magnesium (cMg) concentrations were calculated by substracting ionized fractions from ultrafiltrable fractions. Parathyroid hormone (PTH) and calcitriol (CTR) concentrations were measured via radioimmunoassay. RESULTS: Serum tCa concentration in donkeys (3.37 +/- 0.21 mmol/L) was composed of pCa (1.59 +/- 0.21 mmol/L [47.0 +/- 4.2%]), iCa (1.69 +/- 0.04 mmol/L [50.4 +/- 3.0%]), and cCa (0.09 +/- 0.08 mmol/L [2.6 +/- 2.9%]). Serum tMg concentration (1.00 +/- 0.08 mmol/L) was fractioned in pMg (0.23 +/- 0.08 mmol/L [23.4 +/- 8.1%]), iMg (0.59 +/- 0.04 mmol/L [58.8 +/- 5.1%]), and cMg (0.18 +/- 0.08 mmol/L [17.8 +/- 7.2%]). Serum concentrations of P (1.14 +/- 0.30 mmol/L), PTH (20.4 +/- 21.2 pg/mL), and CTR (13.4 +/- 5.9 pg/mL) were determined. CONCLUSIONS AND CLINICAL RELEVANCE: Serum variables of mineral metabolism in donkeys were within reference ranges for horses. However, when compared with horses, donkeys had higher iCa, cMg, and CTR and lower pMg and PTH concentrations.     

Fractionation of calcium and magnesium in equine serum

OBJECTIVE: To establish reference values for protein-bound, ionized, and weak-acid complexed fractions of calcium and magnesium in equine serum and determine stability of ionized calcium (iCa) and ionized magnesium (iMg) in serum samples kept under various storage conditions. ANIMALS: 28 clinically normal horses. PROCEDURE: Total calcium (tCa) and magnesium (tMg) in equine serum were fractionated by use of a micropartition system that allows separation of protein-bound calcium (pCa) and magnesium (pMg) and ultrafiltrable calcium (microCa) and magnesium (microMg) fractions. Serum concentrations of iCa and iMg were measured in the ultrafiltrate by use of selective electrodes. Serum concentration of complexed calcium (cCa) or magnesium (cMg) was calculated by subtracting iCa or iMg from microCa or microMg, respectively. RESULTS: Mean +/-SE serum tCa concentration was 3.26 +/- 0.06 mmol/L. Calcium fractions were as follows: pCa, 1.55 +/- 0.03 mmol/L (47.4 +/- 0.9%); iCa, 1.58 +/- 0.03 mmol/L (48.5 +/- 0.7%); and cCa, 0.13 +/- 0.02 mmol/L (4.1 +/- 0.9%). Serum tMg concentration was 0.99 +/- 0.04 mmol/L. Magnesium fractions were as follows: pMg, 0.33 +/- 0.04 mmol/L (33.3 +/- 4.2%); iMg, 0.57 +/- 0.02 mmol/L (57.6 +/- 1.7%); and cMg, 0.09 +/- 0.02 mmol/L (9.1 +/- 1.9%). Refrigeration (4 degrees C) did not affect iCa values, whereas iMg declined by 8% after 120 hours. Neither iCa nor iMg was affected by freezing (-20 degrees C). CONCLUSIONS AND CLINICAL RELEVANCE: In equine serum, iMg is less stable than iCa; thus, when serum samples are not going to be analyzed promptly, freezing may be preferable to refrigeration for storage.     

Effects of a supplement containing chromium and magnesium on morphometric measurements, resting glucose, insulin concentrations and insulin sensitivity in laminitic obese horses

Reasons for performing study: Obesity and insulin resistance are risk factors for laminitis in equids and supplements containing chromium and magnesium might improve insulin sensitivity. Hypothesis: A supplement containing chromium, magnesium and other nutraceuticals would alter morphometric measurements, blood variables, and insulin sensitivity in laminitic obese horses. Methods: Twelve previously laminitic obese (body condition score ≥ 7/9) horses were randomly allocated to treatment (n = 6) and control (n = 6) groups and 2 obese horses with clinical laminitis were included in the treatment group. Treated animals received 56 g supplement with 0.25 kg oats once daily for 16 weeks. The supplement contained chromium (5 mg/day as yeast), magnesium (8.8 g/day as oxide/proteinate), and other nutraceuticals. Insulin‐modified frequently sampled i.v. glucose tolerance tests were performed with hay provided at 0, 8 and 16 weeks, and insulin sensitivity was estimated by minimal model analysis. Physical measurements were collected at the same points. Horses were not exercised. Results: Hyperinsulinaemia (>30 µu/ml) was detected in 12 of 14 horses prior to treatment. Glucose and insulin data from one mare with clinical laminitis were excluded because of persistent pain. Mean ± s.d. insulin sensitivity was 0.64 ± 0.62 × 10^?4 l/min/mu prior to treatment for the remaining 13 horses. Time and treatment × time effects were not significant for any of the variables examined, with the exception of resting insulin concentrations, which significantly increased over time (P = 0.018). Health status remained the same. Conclusions: The supplement containing chromium and magnesium evaluated in this study did not alter morphometric measurements, blood variables, resting insulin concentrations or insulin sensitivity in laminitic obese horses. Potential relevance: Additional research is required to determine the appropriate use of chromium and magnesium supplements in horses.     

Equine Metabolic Syndrome

Equine metabolic syndrome (EMS) is important because of its association with laminitis. Obesity and insulin resistance are two important components of EMS, and the underlying cause of this syndrome is likely to be enhanced metabolic efficiency. Affected horses are often referred to as “easy keepers” because they require fewer calories to maintain body condition, and enhanced metabolic efficiency is an inherent risk factor for EMS that may be genetically determined. Pony breeds, Morgan horses, and Paso Finos are predisposed to EMS, but this problem can be prevented through effective management. Overfeeding, abundant pasture grass, and inadequate exercise are risk factors that relate to modern management practices. Obesity and adiposity induce insulin resistance, and recent research suggests that this is the determinant of laminitis susceptibility in ponies. Increased plasma insulin concentrations are detected in most affected horses and ponies, so this serves as a useful screening test for EMS. Physical characteristics also should be examined because horses with EMS exhibit regional adiposity in the form of a cresty neck or abnormal adipose tissue deposits close to the tailhead. All horses with enhanced metabolic efficiency, obesity, or regional adiposity should be screened for EMS. The combined intravenous glucose−insulin test can be performed to diagnose insulin resistance in mildly affected horses and quantify insulin sensitivity. Most horses with EMS can be effectively managed by reducing caloric intake, decreasing the starch and sugar content of the diet, increasing exercise, and limiting or eliminating access to pasture, but medical therapy is warranted in select cases.     

Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses

Equine metabolic syndrome is characterized by obesity and regional adiposity coupled with evidence of recurrent laminitis. Although inflammation has been well characterized in several experimental models of acute laminitis, the inflammatory events associated with endocrinopathic laminitis are not well documented. The aim of this study was to characterize selected markers of inflammation in horses with clinical evidence of equine metabolic syndrome (EMS). Neutrophil phagocytosis and oxidative burst, as well as endogenous and stimulated cytokine expression were evaluated. A marked increase in neutrophil reactive oxygen species production upon phagocytosis was observed in horses with EMS that was strongly correlated to the blood insulin concentration. Increased oxidative burst activity of neutrophils in hyperinsulinemic horses may predispose horses with metabolic syndrome to laminitis. In contrast, peripheral blood cells of obese hyperinsulinemic horses showed decreased endogenous proinflammatory cytokine gene expression (IL-1 and IL-6) and similar cytokine response following immune stimulation compared to that of control horses. This may suggest that, unlike in people, cytokine-mediated inflammation does not increase in direct response to obesity or insulin resistance in horses. This species-specific disparity may explain the difference in clinical outcomes observed in obese horses compared to obese people.     

Glycated Hemoglobin is an Indicator of Blood Glucose Status in Horses: Preliminary Study

Glycated hemoglobin (HbG) concentration is a retrospective measure of mean blood glucose level and is not affected by recent stresses, food ingestion, or exercise. Although HbG has been determined in various wild and domestic animals such as kestrels, mankhor, mouflon, aoudad, deer, goat, sheep, dog, camel, ostrich, and horse, there is no information about diagnostic values of HbG as an indicator of blood glucose status in horses. The purposes of this study were to determine normal value of HbG in Iranian crossbred horses and to investigate its relation to fasting plasma glucose. Blood samples were collected from jugular veins of 193 clinically healthy adult crossbred horses (102 males and 91 females). After separation and washing of red blood cells, hemolysate was prepared and subjected to weak cation exchange chromatography for determination of HbG. Glucose was measured in fasting plasma samples. HbG percent (HbG%) in the studied horses was 3.21 ± 0.56 in males and 3.34 ± 0.72 in females. Fasting plasma glucoses were 81.3 ± 7.6 and 84.2 ± 14.5 mg/dL in males and females, respectively. HbG% and plasma glucose were highly correlated (r = 0.81, P < .01). We concluded that HbG% can be considered as a good indicator of blood glucose status in the horse. Two horses with abnormally higher HbG% were found in this study showing persistent high fasting plasma glucose. We concluded that HbG determination could be a more reliable indicator of basal blood glucose concentrations in horses.     

The Effect of Different Physical Exercise on Plasma Leptin,Cortisol, and Some Energetic Parameters Concentrations in Purebred Arabian Horses

Leptin is a hormone that coordinates food intake, energy expenditure, and metabolic rate; it is, however, unknown how exercise influences plasma leptin concentration in horses. The aim of the study was to evaluate the exercise-induced changes in plasma leptin concentration in purebred Arabian horses competing in races and endurance rides. A total number of 26 horses (12 purebred Arabian racehorses aged 3–5 years and 14 endurance horses aged 7–14 years) were studied during performing routine physical exercise. From each horse, blood samples were collected at rest and immediately after the exercise session. Concentrations of plasma leptin, cortisol, lactic acid, uric acid, triacylglycerols, glycerol and free fatty acids were determined. Exercise-induced increase in cortisol and free fatty acid values were higher in endurance horses than in racehorses. Neither endurance nor race exercises influenced the plasma leptin concentration. Further research is needed to elucidate the effects of different types of exercise on leptin synthesis and secretion in horses.     

Variation of Hematochemical Profile and Vitamin E Status in Feral Giara Horses From Free Grazing in the Wild to Hay Feeding During Captivity

Wildlife protection and management are important priorities for landscape identity and biodiversity preservation. Feeding practices of fauna confined in facilities during temporary captivity are fundamental to support animal health and natural behavior. Appropriate provision of feedstuffs appears to be necessary to support the best practices in respect of animal species-specific natural diet. This investigation explored the variation of the metabolic profile by means of selected metabolite and respective circulating levels in a group feral Giara horses undergoing the change of the diet, moving from natural free grazing in the wild to temporary captivity. Six Giara horses (4 mares and 2 stallions; estimated age: 2.5–3 years; body weight: 163–170 kg) were captured to monitor the serological reaction to equine infectious anemia (EIA; screening at Coggins test). Animals were sheltered in a wildlife rescue center for a duration of 4 weeks, and all received the same hay-based diet (ad libitum). On 0 and 28 days of captivity, blood serum alpha-tocopherol (α-TOH) concentration was determined alongside selected metabolites (liver enzymes, total protein and fractions, cholesterol, triglycerides, and macrominerals and trace elements). Comparative feces quality and composition were also assessed. Both serum samples (0 vs. 28 days) displayed α-TOH levels below (<2 μg/mL) adequacy established for the domestic horse. Initial levels markedly (P = .020) decreased after the 4 weeks of captivity (Δ = −32.5%). Vitamin E status and ALT levels varied significantly, but serum protein fractions did not point to significant variations before and after captivity. All horses tested negative to EIA. Monitoring of vitamin E status of wild and feral herbivores may be recommendable in the context of adequate feeding practices during captivity to prevent potential deficiency or excessive depletion.     

Reference intervals for biochemical and haematological parameters in mature domestic donkeys (Equus asinus) in the UK

Reference intervals (RIs) for haematology and serum biochemistry for donkeys in a temperate climate have been previously published using blood sample results from the resident population of a large donkey shelter in the UK. Periodic review of reference intervals is recommended to ensure their applicability to the patient population and changes in laboratory methods and technologies. The current study aimed to revise the previously published haematology and serum biochemistry values for the adult domestic donkey (Equus asinus) in the UK in the light of a change in analytical equipment at the Donkey Sanctuary laboratory, but also to refine the demography of the sample population with respect to age, physiology and clinical history. Clinical pathology results from 138 clinically healthy mature (4–24 years inclusive) female and castrated male donkeys selected from the resident population of the Donkey Sanctuary, were analysed retrospectively. The animals were blood sampled during the period February 2008 to June 2011 as part of a routine health screen prior to rehoming. Results for a total of 38 biochemical and haematological parameters were analysed including 3 previously unreferenced parameters in addition to those assessed in the previous study. The new reference intervals and median values show very poor transferability with recently derived reference intervals for non‐Thoroughbred horses and only limited transferability with reference intervals previously published for donkeys in the UK. Of particular note is a marked reduction in the upper reference limit for triglycerides of 2.8 mmol/l (from 4.3 mmol/l) since this parameter is used to decide when donkeys are at risk of developing hyperlipaemia. This study demonstrates the value of intermittent review of reference intervals and refinement of study populations. Notwithstanding the caution with which reference interval data from different laboratories should be compared, the lack of transferability of results between donkeys and horses highlights the importance of use of species‐appropriate reference intervals for clinical decision‐making.     

Validation of the Lactate Plus Lactate Meter in the Horse and Its Use in a Conditioning Program

The equine industry has a need for a convenient, rapid, and reliable method of measuring blood lactate concentrations ([LA]). We hypothesized that the handheld Lactate Plus lactate meter (LPlus), developed and tested for use in humans, would provide dependable results when used in horses undergoing an exercise conditioning program and that horse's fitness would improve following individualized conditioning based on each horse's velocity at which [LA] = 4 mmol/L (V_LA4) was reached. Five adult horses underwent a 4-week training program that consisted of 3 exercise bouts/wk. Horses were subjected to an incremental step standardized exercise test (SET) before starting (SET-1) and after the completion of the program (SET-2). Blood samples were collected before each increase in speed until [LA] reached ≥4 mmol/L, and then the SET was terminated. The [LA] sample range in our study was 0–8 mmol/L. Blood was analyzed at the time of collection using a calibrated LPlus, and plasma was collected for [LA] determination using the lactate dehydrogenase–based enzymatic colorimetric method. Although the LPlus tended to significantly underestimate [LA] by 0.39 mmol/L (P < .001), the LPlus proved to be a dependable device for use in horses based on good correlation with the biochemical analysis (r = 0.978) and Bland–Altman limits of agreement and 95% confidence intervals. All horses showed an increase in V_LA4 from SET-1 to SET-2, consistent with improved fitness following our 3 exercise bout/wk training protocol. The LPlus can reliably be used in horses to determine [LA] ranging from 0–8 mmol/L. When determining serial [LA], analytical techniques should not be used interchangeably.     

Circadian rhythm of calciotropic hormones,serum calcium,phosphorus and magnesium during the shortest and longest days of the year in horses in New Zealand

A study was conducted to determine the circadian rhythms and trends of vitamin D metabolites including 25‐hydroxyvitamin D₃, 25‐hydroxyvitamin D₂, 1,25‐dihydroxyvitamin D and parathyroid hormone, in addition to serum calcium, phosphorus and magnesium concentrations in horses over 48 h on the shortest and longest days of the year in 2013. Five healthy adult horses (Equus caballus) were on a constant pasture feeding regimen, and blood samples were collected from each horse every 3 h over a 48‐h period, starting at 07:00 PM on day one and finishing at 07:00 PM on day three, for the measurement of calciotropic hormones and electrolytes. There was a significant difference between the serum concentration of calciotropic hormones, iCa, tCa, P and tMg between the shortest (winter) and longest (summer) days of the year in horses. Serum concentration of 25OHD₃ was very low and mostly undetectable. Serum iCa, 1,25(OH)₂D and PTH concentrations clearly showed a circadian rhythm on the longest days of the year and serum tCa, P and tMg concentrations showed a diurnal pattern on the longest days (summer) of the year. None of the analytes showed any circadian rhythm on the shortest days (winter) of the year. The result of this study could have significant relevance to equine athletes travelling to international equestrian competitions and facing a huge time and seasonal differences that might affect their ability to adjust their circadian rhythms to new time zones.     

Interday variation and effect of transportation on indirect blood pressure measurements,plasma endothelin-1 and serum cortisol in Standardbred and Icelandic horses

Background

Systemic hypertension is a prominent feature in humans with metabolic syndrome (MS) and this is partly caused by an enhanced endothelin-1 (ET-1) mediated vasoconstriction. There are indications that systemic hypertension might be a feature in equine metabolic syndrome (EMS) but if ET-1 is involved in the development of hypertension in horses is not known. Increased levels of cortisol have also been found in humans with MS but there are no reports of this in horses. Before blood pressure, plasma ET-1 and serum cortisol can be evaluated in horses with EMS, it is necessary to investigate the interday variation of these parameters on clinically healthy horses. The aims of the present study were therefore to evaluate the interday variation and influence of transportation on systemic blood pressure, plasma ET-1 and serum cortisol in healthy Standardbred and Icelandic horses, and to detect potential breed differences.

Methods

Nine horses of each breed were included in the study. Blood pressure was measured and blood samples were collected between 6 and 9 am on two separate days. Eight of the horses (four of each breed) were transported to a new stable were they stayed overnight. The next morning, the sampling procedure was repeated.

Results

The interday variation was higher for plasma ET-1 (37%) than for indirect pressure measurements (8-21%) and serum cortisol (18%). There were no differences in systemic blood pressure between the two breeds. The Icelandic horses had significantly lower serum cortisol and significantly higher plasma ET-1 concentrations compared to the Standardbred horses. Plasma ET-1 was significantly elevated after transportation, but systemic blood pressure and serum cortisol did not differ from the values obtained in the home environment.

Conclusions

Indirect blood pressure, plasma ET-1 and serum cortisol are of interest as markers for cardiovascular dysfunction in horses with EMS. The elevated plasma ET-1 concentrations recorded after transportation was likely caused by a stress response. This needs to be considered when evaluating plasma ET-1 in horses after transportation. The differences detected in plasma ET-1 and serum cortisol between the two breeds might be related to differences in genetic setup, training status as well as management conditions.     

Optimum conditions for serum clearance of iodixanol,applicable to the estimation of glomerular filtration rate in horses

To estimate the glomerular filtration rate (GFR) in horses, an optimum dose of the nonionic contrast medium iodixanol as a tracer was assessed with blood-sample times. Iodixanol was administered intravenously at 10–40 mg I/kg to geldings and mares, and blood was collected 30, 60, 90, 120, 150, and 180 min later. Serum iodixanol concentration was determined by high-performance liquid chromatography (HPLC), and serum urea nitrogen (UN) and creatinine concentrations were also measured. The combination of 20 mg I/kg iodixanol and sampling times of 60, 90, and 120 min after injection was considered to be appropriate for practical use. In clinically healthy horses, the reference values were determined to be 1.90 ± 0.03 ml/min/kg (150.8 ± 2.94 ml/min/m²), consistent with historical data using different tracers. The result suggests that serum clearance of iodixanol is a ready-to-use tool for a screening of alterations in the equine GFR, although it is necessary to perform a more longitudinal study using horses with a variety of renal functions.