Endocrine, morphometric, and ultrasonographic characterization of neck adiposity in Andalusian horses.

Equine metabolic syndrome (EMS) can be diagnosed by hormonal measurements; however, it would be important to find simpler measurements that allow easy identification of affected or at risk individuals. In horses, the dorsal neck region is one of the most frequent anatomical sites for fat deposition and neck obesity has been linked to EMS. The aim of this study was to evaluate the association of hormonal markers of obesity (leptin) and insulin resistance (insulin) with morphometric and ultrasonographic neck measurements in Andalusian horses. Plasma leptin and insulin concentrations were measured by RIA in 127 Andalusian horses. Neck circumferences (NC) were measured at 3 equidistant locations at 25%, 50%, and 75% of neck length (NC-25%, NC-50%, and NC-75%). At the same 3 locations, subcutaneous fat thickness (SFT-25%, SFT-50%, and SFT-75%) was measured ultrasonographically. In the population under study, a tendency to adiposity was confirmed by the elevated plasma leptin levels (7.47 ± 5.03 ng/mL). However, plasma insulin concentrations (4.05 ± 3.74 µIU/mL) were within normal range in most horses. Our results indicate that NC showed significant sexual dimorphism and did not correlate well with hormonal measurements. Ultrasonographic assessment of fat thickness at the base of the neck (SFT-75%) was significantly correlated with both plasma leptin and insulin and did not show differences between males and females. Thus, in the search for a single objective parameter which can be used in large populations, SFT-75% is a potential candidate and may be a meaningful parameter to predict EMS.

Eosinophilic Enteritis in Horses with Motor Neuron Disease.

BACKGROUND: Equine motor neuron disease (EMND) is a neuromuscular disorder that affects adult horses. Although EMND has been linked to vitamin E deficiency, its etiopathogenesis is poorly understood. OBJECTIVES: To describe clinical features, laboratory results, and postmortem findings in a series of young horses with motor neuron disease (MND). ANIMALS: A herd of 15 young Andalusian horses with weakness, weight loss, muscle atrophy, and muscle fasciculations related to restricted intake of green forage. METHODS: A case series is presented in which horses were subjected to a clinical examination and plasma vitamin E measurement. Five severely affected horses were euthanized for detailed postmortem examination. Muscle specimens were taken from the M. sacrocaudalis dorsalis medialis and the M. gluteus medius for histopathologic and morphometric evaluation. RESULTS: MND was diagnosed in 5 horses based on clinical signs, low serum levels of vitamin E (0.11 ± 0.05 mg/dL; normal range,: 0.3-1.5 mg/dL), changes in muscle histopathology (neurogenic atrophy), and spinal cord lesions (neuronal chromatolysis in ventral horns). An unexpected postmortem finding was the presence of intestinal inflammation (catarrhal enteritis, edema, and eosinophilic infiltrate) associated with the presence of giant ciliated protozoa in all of the horses. CONCLUSIONS: Although a mechanistic link could not be established, it is hypothesized that intestinal inflammation may have been involved in the decreased absorption of vitamin E, thus favoring the development of MND.

Effects of two calculolytic diets on parameters of feline mineral metabolism.

OBJECTIVES: To evaluate the influence of two feline calculolytic diets on selected parameters of mineral metabolism. MATERIALS AND METHODS: Two dry commercial diets designed for struvite urolith dissolution were evaluated in 14 cats. The study was designed as a two-sequence, four-period crossover protocol with a baseline period, two 60-day "run-in" periods in which calculolytic diets (Diet 1 and Diet 2) were fed and one 30-day "wash-out" period. Data are expressed as median (range). RESULTS: Feeding the calculolytic diets for two months did not alter plasma concentrations of calcium, phosphorus, magnesium and parathyroid hormone. A significant (P < 0.05 in each case) decline in calcitriol was observed after administering both diets from 236.4 (122.4-429.6) to 170.4 (108.0-394.3) pmol/L (Diet 1) and from 278.4 (153.6-492.0) to 177.1 (87.6-392.4) pmol/L (Diet 2). Cats fed Diet 1 showed a significant increase in urine calcium concentration (from 0.3 (0.2-0.5) to 0.4 (0.3-0.7) mmol/L). Magnesium concentration in urine was significantly increased with both diets, from 1.4 (0.1-1.7) to 1.5 (1.3-2.4) mmol/L (Diet 1) and from 1.1 (0.4-1.9) to 2.0 (0.1-3.1) mmol/L (Diet 2). CLINICAL SIGNIFICANCE: Both diets resulted in an increased urinary concentration of magnesium, through different mechanisms: urine acidification (Diet 1) and increased sodium load (Diet 2).

Repeated measurements of P retention in ponies fed rations with various Ca:P ratios.

This study addresses the question of whether feeding rations rich in P for a period of up to 42 d induces a positive P balance in adult ponies. Biochemical bone markers and parathyroid hormone (PTH; intact as well as whole PTH) were measured to obtain clues as to the effect of P loading on bone metabolism. The experiment had a Latin square design. Each feeding period lasted 42 d, and there were 2 balance trials (ECP1 and ECP2) within each feeding period. Each balance trial lasted 10 d (ECP1: d 11 to 21; ECP2: d 33 to 42). Six ponies aged 2.5 to 7 yr were fed a control diet that provided P and Ca according to the requirement (Control diet: 54 mg Ca·kg BW(-1) · d(-1); 36 mg P · kg BW(-1) · d(-1)), a diet high in Ca and P (HCaHP diet: 146 mg Ca · kg BW(-1) · d(-1); 121 mg P · kg BW(-1) · d(-1)), and a diet with a high P level only and Ca fed to the requirement (HP diet: 54 mg Ca · kg BW(-1) · d(-1); 122 mg P · kg BW(-1) · d(-1)). When fed the Control diet, the ponies showed a zero P and Ca balance over the 42-d period. The HCaHP diet resulted in both P and Ca retention (about 2 g Ca and P/d; P < 0.05). Phosphorus retention (about 2 g P/d) alone was observed when ponies were fed the HP diet, but P retention was only different (P < 0.05) from the Control diet in ECP1. The excretion of P in urine was reduced by greater Ca intake (P < 0.05), and Mg absorption was reduced by high P intake (P < 0.05). Plasma P concentration was raised by high P intake. Plasma Ca levels were not affected by dietary treatment. The greater (P < 0.05) P retentions observed for the HCaHP diet during ECP1 and ECP2 and HP diet during ECP1 could not be explained by processes that could have been indicated by the bone markers or PTH values. It was concluded that dietary-P-induced retention of P in ponies does not seem to be associated with altered bone metabolism in this study.

Influence of feeding status, time of the day, and season on baseline adrenocorticotropic hormone and the response to thyrotropin releasing hormone-stimulation test in healthy horses.

Equine pituitary pars intermedia function can be assessed by the measurement of baseline and thyrotropin releasing hormone (TRH)-induced concentrations of adrenocorticotropic hormone (ACTH); however, these measurements may be affected by the environment. Therefore, a prospective observational study evaluated the influence of feeding, time of the day, and season on baseline and TRH-induced concentrations of ACTH in healthy horses. Baseline ACTH was measured in 50 horses before and 2 h after feeding. Six research horses were subjected to a crossover study in which 6 TRH tests were performed in 2 different seasons, March-April (MA) and July-September (JS), at 2 different times of the day, 8 AM and 8 PM, and, under 2 different conditions relative to feeding status, fasted and 2 h after feeding. Differences between fasted and fed horses were found in baseline ACTH, 17.1 ± 1.8 versus 46.1 ± 7.6 pg/mL (P = 0.003) and TRH-stimulated ACTH: 124.1 ± 21.3 versus 192.6 ± 33.1 pg/mL (P = 0.029) at 10 min, and 40.1 ± 4.9 versus 73.2 ± 13.4 pg/mL (P = 0.018) at 30 min post TRH injection. No differences were found between tests performed at different times of the day. Basal ACTH concentrations were greater in JS than in MA, 17.1 ± 1.8 versus 11.9 ± 0.6 pg/mL (P = 0.006). A seasonal influence was also found in stimulated ACTH values, which were much greater in JS 122.7 ± 36.7 versus 31.2 ± 7.4 pg/mL, at 10 min (P = 0.03) and 39.0 ± 7.2 versus 19.8 ± 3.1 pg/mL, at 30 min (P = 0.03). In addition to season, feeding is a potential confounding factor when measuring baseline or stimulated ACTH in horses. In conclusion, feeding status should be standardized for the diagnosis of equine pituitary pars intermedia dysfunction.

Vitamin E protection of obesity-enhanced vascular calcification in uremic rats.

This study aimed to determine the extent of extraskeletal calcification in uremic Zucker rats, by comparing obese and lean phenotypes, and to evaluate the influence of vitamin E (VitE) on the development of calcifications in both uremic rats and human vascular smooth muscle cells (HVSMCs) cultured in vitro. Zucker rats of lean and obese phenotypes with normal renal function [control (C); C-lean and C-obese groups] and with uremia [5/6 nephrectomy (Nx); Nx-lean and Nx-obese groups] and uremic rats treated with VitE (Nx-lean + VitE and Nx-obese + VitE groups) were studied. Uremic groups were subjected to Nx, fed a 0.9% phosphorus diet, and treated with calcitriol (80 ng/kg ip). The aortic calcium concentration was significantly higher (P < 0.05) in Nx-obese rats (10.0 ± 2.1 mg/g tissue) than in Nx-lean rats (3.6 ± 1.3 mg/g tissue). A decrease in plasma glutathione peroxidase activity was observed in Nx-obese rats compared with Nx-lean rats (217.2 ± 18.2 vs. 382.3 ± 15.5 nmol·min(-1)·ml(-1), P < 0.05). Treatment with VitE restored glutathione peroxidase activity and reduced the aortic calcium concentration to 4.6 ± 1.3 mg/g tissue. The differences in mineral deposition between Nx-lean, Nx-obese, Nx-lean + VitE, and Nx-obese + VitE rats were also evidenced in other soft tissues. In HVSMCs incubated with high phosphate, VitE also prevented oxidative stress and reduced calcium content, bone alkaline phosphatase, and gene expression of core-binding factor-α1. In conclusion, uremic obese rats develop more severe calcifications than uremic lean rats and VitE reduces oxidative stress and vascular calcifications in both rats and cultures of HVSMCs.

Comparison of different treatments for oestrous induction in seasonally anovulatory mares.

The aim of this study was to evaluate the effects of different treatments for induction and synchronization of oestrus and ovulation in seasonally anovulatory mares. Fifteen mares formed the control group (C), while 26 mares were randomly assigned to three treatment groups. Group T1 (n = 11) were treated with oral altrenogest (0.044 mg/kg; Regumate(®) ) during 11 days. Group T2 (n = 7) was intravaginally treated with 1.38 g of progesterone (CIDR(®) ) for 11 days. In group T3 (n = 8), mares were also treated with CIDR(®) , but only for 8 days. All mares received PGF2α 1 day after finishing the treatment. Sonographic evaluation of follicles, pre-ovulatory follicle size and ovulation time was recorded. Progesterone and leptin levels were analysed. Results show that pre-ovulatory follicles were developed after the treatment in 88.5% of mares. However, the pre-ovulatory follicle growth was dispersal, and sometimes it was detected when treatment was not finished. While in mares treated with intravaginal device, the follicle was soon detected (1.5 ± 1.2 days and 2.3 ± 2.0 days in T2 and T3 groups, respectively), in T1 group, the pre-ovulatory follicle was detected slightly later (3.9 ± 1.6 days). The interval from the end of treatment to ovulation did not show significant differences between groups (T1 = 13.1 ± 2.5 days; T2 = 11.0 ± 3.6 days; T3 = 13.8 ± 4.3 days). The pregnancy rate was 47.4%, similar to the rate observed in group C (46.7%; p > 0.05). Initial leptin concentrations were significantly higher in mares, which restart their ovarian activity after treatments, suggesting a role in the reproduction mechanisms in mares. It could be concluded that the used treatments may be effective for oestrous induction in mares during the late phase of the seasonally anovulatory period. Furthermore, they cannot synchronize oestrus, and then, it is necessary to know the reproductive status of mares when these treatments are used for oestrous synchronization.

Plasma leptin concentration in donkeys.

BACKGROUND: Donkeys appear to be more predisposed than large breed horses to suffer from hyperlipemia. The reason for that predisposition is unknown but anorexia is a consistent feature of the disease. Leptin, a protein synthesized in fat tissue, is one of the major inhibitors of appetite in mammals. OBJECTIVE: We hypothesized that donkeys could have elevated plasma leptin concentrations compared to horses. ANIMALS AND METHODS: Blood samples were obtained from 50 donkeys for measurement of leptin, triglycerides (TGs), glucose, and insulin. Glucose/insulin ratio, modified insulin to glucose ratio, and reciprocal of the square root of insulin were calculated. Based on their body condition score (BCS), donkeys were classified as lean (n = 18), normal (n = 16), or overweight (n = 16). The results were compared with reference values from our laboratory and with a group of horses (n = 25) used as an internal control. RESULTS: Values of both leptin and TGs in donkeys were above the horse reference range and also significantly higher than those of the control horses: leptin (11.2 ± 1.7 versus 5.8 ± 0.5 µg/L, p < 0.05) and TGs (0.93 ± 0.1 versus 0.54 ± 0.1 mmol/L, p < 0.01). Overweight donkeys had leptin (19.3 ± 2.9 µg/L) and TG (1.3 ± 0.2 mmol/L) concentrations that were significantly (p < 0.01) higher than normal (9.4 ± 3.3 µg/L and 0.85 ± 0.1 mmol/L, respectively) and lean (5.5 ± 1.0 µg/L and 0.66 ± 0.1 mmol/L, respectively) donkeys. A significant positive correlation (p < 0.01) was found between BCS and leptin (r = 0.43), TGs (r = 0.46), glucose (r = 0.41), and insulin (r = 0.40). CONCLUSION: Donkeys have higher plasma leptin concentrations than horses and leptin is correlated with BCS.

Feline parathyroid hormone: validation of hormonal assays and dynamics of secretion.

Validated assays for quantification of intact parathyroid hormone (I-PTH) are no longer available. Moreover, the third-generation PTH assay that only detects the whole PTH molecule (W-PTH) has never been tested in cats. The work presented here is aimed to validate a commercially available assay for measurement of I-PTH and W-PTH in cats and to study the dynamics of PTH secretion in healthy cats. Our results show that both assays are reliable for the measurement of feline PTH. In healthy adult cats W-PTH concentration (15.1 ± 1.6 pg/mL) was greater (P < 0.001) than I-PTH concentration (9.1 ± 0.7 pg/mL). The dynamics of PTH secretion in response to changes in extracellular calcium (Ca(2+)) were investigated in 13 cats by studying PTH-Ca(2+) curves. PTH-Ca(2+) curves were obtained by intravenous infusion of disodium ethylenediaminetetraacetic acid and CaCl(2). PTH was measured using both I-PTH and W-PTH assays. During hypocalcemia a sigmoidal curve that was similar when measured with I-PTH or W-PTH was obtained. The maximal PTH concentration in response to hypocalcemia was greater with W-PTH (179.6 ± 41.9 pg/mL) than with I-PTH (67.6 ± 10.5 pg/mL; P = 0.01). However, hypercalcemia resulted in an equivalent PTH inhibition, with both assays yielding PTH concentrations as follows: W-PTH = 4.0 ± 0.4 pg/mL and I-PTH = 4.9 ± 0.3 pg/mL (NS). Parameters of the feline PTH-Ca(2+) curve are similar to what has been previously reported in dogs.

Metabolic acidosis inhibits soft tissue calcification in uremic rats.

Metabolic acidosis is common in patients with chronic kidney disease, which is known to affect bone metabolism. We examined the effect of metabolic acidosis on the development of vascular and other soft-tissue calcifications in uremic rats treated with calcitriol. Extraskeletal calcification was measured in vivo, in control rats and rats with a remnant kidney model of uremia with or without ammonium chloride-induced acidosis. Soft-tissue calcification was assessed histologically, by measurement of the expression of the sodium-dependent phosphate cotransporter Pit-1 and by quantification of tissue calcium and phosphorus. Calcitriol administration to uremic rats resulted in significant deposition of material positive for von Kossa stain in the aorta, stomach, and kidney, elevated aortic calcium and phosphorus, increased aortic Pit-1 expression, and high mortality. Calcitriol-treated uremic rats with acidosis did not develop aortic or soft-tissue calcification, did not increase aortic Pit-1 expression, and had significantly lower mortality. Additionally, an acidotic environment prevented calcification of vascular smooth muscle cells in vitro. Our study shows that metabolic acidosis inhibits extraskeletal calcification.

The effect of calcitriol, paricalcitol, and a calcimimetic on extraosseous calcifications in uremic rats.

Vitamin D derivatives and calcimimetics are used to treat secondary hyperparathyroidism in patients with chronic renal failure. We investigated the effect of calcitriol, paricalcitol, and the calcimimetic AMG 641 on soft-tissue calcification in uremic rats with secondary hyperparathyroidism. Control and uremic rats were treated with vehicle, calcitriol, paricalcitol, AMG 641, or a combination of AMG 641 plus calcitriol or paricalcitol. Parathyroid hormone levels were reduced by all treatments but were better controlled by the combination of paricalcitol and AMG 641. The calcimimetic alone did not induce extraosseous calcification but co-administration of AMG 641 reduced soft-tissue calcification and aortic mineralization in both calcitriol- and paricalcitol-treated rats. Survival was significantly reduced in rats treated with calcitriol and this mortality was attenuated by co-treatment with AMG 641. Our study shows that extraskeletal calcification was present in animals treated with calcitriol and paricalcitol but not with AMG 641. When used in combination with paricalcitol, AMG 641 provided excellent control of secondary hyperparathyroidism and prevented mortality associated with the use of vitamin D derivatives without causing tissue calcification.

Circadian variation of the Ca2+ -PTH curve during hypercalcaemia in dogs.

To test the hypothesis that the parathyroid hormone (PTH) response to hypercalcaemia is influenced by circadian rhythms, the Ca2+ -PTH curve was studied in six dogs after infusion of CaCl2 (0.66 mEq/kg/h) at daytime (09:00 h) and at night-time (21:00 h). Plasma Ca2+ and PTH values measured before or after CaCl2 infusion were not different at day and at night. However, in the recovery from hypercalcaemia, PTH concentrations were significantly lower (P < 0.05) at 21:00 h (23 +/- 7.5 pg/ml at Ca2+ = 1.30 mm) than at 09:00 h (38.8 +/- 6.9 pg/ml at Ca2+ = 1.30 mm). In addition, the Ca2+ -PTH curve showed hysteresis at daytime (for the same Ca2+ concentration, PTH values were higher during recovery than during induction of hypercalcaemia) but not at night-time (PTH values were lower during recovery than during induction of hypercalcaemia). In conclusion, a circadian variation in the PTH secretory pattern during recovery from hypercalcaemia has been identified in dogs.

The calcimimetic R-568 increases vitamin D receptor expression in rat parathyroid glands.

We previously demonstrated that extracellular calcium regulates vitamin D receptor (VDR) expression by parathyroid cells. Since the calcimimetic R-568 potentiates the effects of calcium on the calcium-sensing receptor, it was hypothesized that administration of R-568 may result in increased VDR expression in parathyroid tissue. In vitro studies of the effect of R-568 on VDR mRNA and protein were conducted in cultures of whole rat parathyroid glands and human hyperplastic parathyroid glands. In vivo studies in Wistar rats examined the effect of R-568 and calcitriol alone and in combination. Incubation of rat parathyroid glands in vitro with R-568 (0.001-1 microM) resulted in a dose-dependent decrease in parathyroid hormone (PTH) secretion and an increase in VDR expression (mean +/- SE). Incubation in 1 mM calcium + 0.001 microM R-568 elicited an increase in VDR mRNA (306 +/- 46%) similar to the maximum increase detected with 1.5 mM calcium (330 +/- 42%). In vivo, VDR mRNA was increased after administration of R-568 (168 +/- 9%, P < 0.001 vs. control) or calcitriol (198 +/- 16%, P < 0.001 vs. control). Treatment with R-568 also increased VDR protein in normal rat parathyroid glands and in human parathyroid glands with diffuse, but not nodular, hyperplasia. In conclusion, the present study shows that the calcimimetic R-568 exerts a stimulatory effect on VDR expression in the parathyroid glands of study models and provides additional evidence for the use of calcimimetics in the treatment of secondary hyperparathyroidism.

An unusual case of generalized soft-tissue mineralization in a suckling foal.

An atypical case of severe soft-tissue mineralization in a 3-week-old foal from a herd of Andalusian horses is described. The herd clinical history and the laboratory findings were compatible with a diagnosis of secondary hyperparathyroidism due to a mineral imbalance in the diet (low calcium and high phosphorus intake). Mares showed a marked increase in serum parathyroid hormone (PTH) approximately 10 times normal levels. Serum PTH was marginally elevated in foals. Clinical signs (unthriftiness, painful joints, lameness in one or more limbs, and stiff gait) were more pronounced in foals than in mares. Two foals died and necropsy of one of them revealed extensive soft-tissue mineralization of arterial walls and pulmonary parenchyma. Clinical signs in mares and foals resolved by 4 weeks after diet adjustment.

The influence of anticoagulants on the measurement of total protein concentration in equine peritoneal fluid.

The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The solution of K3EDTA in distilled water only caused TP overestimation at high K3EDTA concentrations (>5 micromol/ml). By contrast, lithium heparin did not influence the refractometric values of TP. Neither anticoagulant modified TP values when measured by the biuret method. In conclusion, the use of K3EDTA as anticoagulant may result in a significant overestimation of TP values of peritoneal fluid samples measured by refractometry.

Diurnal variations in the plasma concentration of parathyroid hormone in dogs.

The plasma concentrations of parathyroid hormone (PTH), ionised calcium (Ca(2+)), total calcium, albumin and inorganic phosphorus, and the pH were measured in blood samples obtained from nine dogs during a period of 26 hours. The plasma pth levels fluctuated slightly during the day, by about 20 pg/ml, but there was a distinct peak (42.8 [8.8] pg/ml) at 07.00. Plasma Ca(2+) showed a diurnal pattern in which two peaks (increases of 0.03 mmol/l) were observed at 05.00 and 17.00, and the plasma concentration of inorganic phosphorus showed a similar pattern. There were no diurnal changes in total calcium or albumin.

The influence of the progression of secondary hyperparathyroidism on the set point of the parathyroid hormone-calcium curve.

The influence of secondary hyperparathyroidism (2 HPT) on the set point of the parathyroid hormone (PTH)-Ca(2+) curve is controversial. In vitro experiments have shown an increase in the set point. However, clinical studies with hemodialysis patients have provided a variety of results (increases, decreases and no changes in the set point have been reported). The present study was designed to investigate the influence of the progression of 2 HPT on the set point of the PTH-Ca(2+) curve. The PTH-Ca(2+) curve and the expression of parathyroid calcium receptor (CaR mRNA) and vitamin D receptor (VDR mRNA) have been studied in normal rabbits (group I, n=9) and in nephrectomized rabbits (group II, n=18) at two stages after inducing 2 HPT: 2-3 weeks (group IIA) and 5-6 weeks (group IIB). In group I, the set point of the PTH-Ca(2+) curve was 1.63+/-0.03 mM. A progressive hypocalcemia was detected during the evolution of 2 HPT (groups IIA and IIB). Rabbits from group IIA had a significant (P<0.001) decrease in the set point to values of 1.45+/-0.02 mM. However, the set point increased significantly in group IIB (P<0.001) to 1.56+/-0.03 mM. CaR mRNA was similarly decreased in groups IIA (39+/-12%) and IIB (48+/-7%). No changes were detected in VDR mRNA. In conclusion, a reduction in the set point of the PTH-Ca(2+) curve in response to decreased extracellular Ca(2+) was detected in the early phases of 2 HPT. However, with the progression of 2 HPT the set point tended to increase even though extracellular Ca(2+) was markedly decreased. The increase in the set point in the course of 2 HPT seems to be a complex process that cannot be fully explained by changes in parathyroid CaR mRNA or VDR mRNA.

Validation and clinical utility of a novel immunoradiometric assay exclusively for biologically active whole parathyroid hormone in the horse.

REASONS FOR PERFORMING STUDY: Parathyroid hormone (PTH) plays a critical role in the regulation of mineral metabolism in mammals. Until recently, the standard method for PTH measurement has been the 2nd generation intact-PTH (I-PTH) assay. Current evidence indicates that the I-PTH assay binds to the PTH molecule and to an inactive N-terminally truncated PTH fragment that tends to accumulate in the blood of uraemic patients. Therefore, a new 3rd generation PTH assay that detects only the whole PTH molecule (W-PTH; cyclase-activating PTH [CAP]) has been developed. OBJECTIVES: To validate this more specific W-PTH assay for measurement of equine PTH and evaluate its clinical utility. METHODS: W-PTH and I-PTH were measured in plasma samples from normal horses (adults and foals) and horses with nutritional secondary hyperparathyroidism (N2HPT) and with chronic renal failure (CRF). Replicate measurements and dilutional paralellism were used for assay validation. Changes in blood ionized calcium were induced by EDTA and CaCl2 administration. RESULTS: Performance of the W-PTH assay (accuracy, sensitivity, specificity and ability to detect changes in PTH in response to changes in calcium) was similar to that of the I-PTH assay. Surprisingly, the relative W-PTH concentration in normal horses and foals was higher than the relative I-PTH concentration. W-PTH values remained higher than I-PTH during acute hypo- and hypercalcaemia. An increase in both W-PTH and I-PTH concentrations was found in horses with N2HPT. In horses with CRF, W-PTH and I-PTH values were very low and no increase in I-PTH was observed. CONCLUSIONS: The W-PTH assay can be used for measurement of equine PTH. POTENTIAL RELEVANCE: The use of W-PTH assay is likely to improve the diagnosis of mineral metabolism in horses.