Phenotypic, hormonal, and clinical characteristics of equine endocrinopathic laminitis

Background Equine endocrinopathic laminitis is common and can be associated with an underlying endocrinopathy, such as equine metabolic syndrome (EMS), pituitary pars intermedia dysfunction (PPID), pasture consumption, or any combination of these factors. Objectives The aim of the study was to improve the risk assessment capabilities of clinicians, and to inform management strategies, for acute endocrinopathic laminitis by prospectively examining the phenotypic, hormonal, and clinical characteristics of the disease in a large cohort. Animals Privately owned horses and ponies (n = 301) of any age, sex, or breed diagnosed with laminitis by a veterinarian. A history of laminitis was acceptable. Methods This was a prospective cohort study. Veterinarians provided information on each case via an online questionnaire after informed consent from the animal's owner, and all data were de‐identified before analysis. Serum insulin and plasma adrenocorticotrophic hormone concentrations were measured in each case. Results Most cases were recruited in spring (109/301; 36.2%). Concurrent EMS and PPID resulted in higher basal insulin concentrations (49 [21.5‐141]; P < .02) than if an animal had a single underlying cause for their laminitis. The insulin concentration was negatively correlated (r2 = −0.38; P < .001) with the animal's height, being higher in ponies (33[10‐14]; P < .001) than horses (9.5 [3‐25.7]) and was positively correlated (r2 = 0.12; P = .05) with their grade (severity) of laminitis. Conclusions and Clinical Importance Horses and ponies with concurrent endocrinopathies have more marked hyperinsulinemia. Higher basal insulin concentrations were associated with more severe lameness

Incidence and risk factors for recurrence of endocrinopathic laminitis in horses

Abstract Background Endocrinopathic laminitis is common in horses and ponies, but the recurrence rate of the disease is poorly defined. Objectives To determine the incidence of, and risk factors for, the recurrence of endocrinopathic laminitis. Animals Privately owned horses and ponies with acute laminitis (n = 317, of which 276 cases with endocrinopathic laminitis were followed up to study completion). Methods This prospective cohort study collected data on veterinary‐diagnosed cases of acute laminitis for 2 years. Each case was classified on acceptance to the study as endocrinopathic or non‐endocrinopathic using data collected in a questionnaire completed by the animal's veterinarian. Follow‐up data were collected at regular intervals to determine whether the laminitis recurred in the 2‐year period after diagnosis. Results The recurrence rate for endocrinopathic laminitis was 34.1%. The risk of recurrence during the 2‐year study period increased with basal, fasted serum insulin concentration (P ≤ .05), with the probability of recurrence increasing markedly as the insulin concentration increased beyond the normal range (0‐20 μIU/mL) to over the threshold for normal (up to approximately 45 μIU/mL). Being previously diagnosed with laminitis (before the study; P = .05) was also a risk factor for recurrent laminitis. Cases with a higher Obel grade of laminitis were likely (P = .05) to recur sooner. Conclusions and clinical importance Knowing that hyperinsulinemia and being previously diagnosed with laminitis are significant risk factors for recurrence will enable clinicians to proactively address these factors, thereby potentially reducing the risk of recurrence of laminitis

Metabolic profile distinguishes laminitis-susceptible and -resistant ponies before and after feeding a high sugar diet

Background: Insulin dysregulation (ID) is a key risk factor for equine endocrinopathic laminitis, but in many cases ID can only be assessed accurately using dynamic tests. The identification of other biomarkers could provide an alternative or adjunct diagnostic method, to allow early intervention before laminitis develops. The present study characterised the metabolome of ponies with varying degrees of ID using basal and postprandial plasma samples obtained during a previous study, which examined the predictive power of blood insulin levels for the development of laminitis, in ponies fed a high-sugar diet. Samples from 10 pre-laminitic (PL – subsequently developed laminitis) and 10 non-laminitic (NL – did not develop laminitis) ponies were used in a targeted metabolomic assay. Differential concentration and pathway analysis were performed using linear models and global tests. Results: Significant changes in the concentration of six glycerophospholipids (adj. P ≤ 0.024) and a global enrichment of the glucose-alanine cycle (adj. P = 0.048) were found to characterise the response of PL ponies to the high-sugar diet. In contrast, the metabolites showed no significant association with the presence or absence of pituitary pars intermedia dysfunction in all ponies. Conclusions: The present results suggest that ID and laminitis risk are associated with alterations in the glycerophospholipid and glucose metabolism, which may help understand and explain some molecular processes causing or resulting from these conditions. The prognostic value of the identified biomarkers for laminitis remains to be investigated in further metabolomic trials in horses and ponies.</p

A "modified Obel" method for the severity scoring of (endocrinopathic) equine laminitis

Background: Laminitis is a common equine disease characterized by foot pain, and is commonly diagnosed using a five-grade Obel system developed in 1948 using sepsis-related cases. However, endocrinopathic laminitis is now the most common form of the disease and clinical signs may be mild, or spread across two Obel grades. This paper describes a modified method which assigns scores to discreet clinical signs, providing a wider scale suitable for use in a research setting.Methods: The "modified Obel" method was developed using an iterative process. First, a prototype method was developed during the detailed observation of 37 ponies undergoing a laminitis induction experiment. The final method was refined and validated using video footage taken during the induction study and from a clinical trial of naturally occurring endocrinopathic laminitis cases. The Obel method was deconstructed and key laminitis signs were evaluated to develop a three-stage, five criteria method that employs a severity scale of 0-12. Veterinarians (n = 28) were recruited to watch and assess 15 video recordings of cases of varying severity, using the Obel and "modified Obel" methods. The inter-observer agreement (reproducibility) was determined using Kendall's coefficient of concordance (Kendall W) and Krippendorf's alpha reliability coefficient. A total of 14 veterinarians repeated the exercise 2-4 weeks after their original assessment, to determine intra-observer agreement (repeatability), assessed using a weighted kappa statistic (kw). Agreement between methods was calculated by converting all "modified Obel" scores to Obel grades and calculating the mean and distribution of the differences.Results: The "modified Obel" and Obel methods showed excellent and similar inter-observer agreement based on the Kendall W value (0.87, P < 0.001 vs. 0.85, P < 0.001) and Krippendorfs alpha (95% CI) value (0.83 [0.53-0.90] vs. 0.77 [0.55-0.85]). Based on the kw value, the "modified Obel" method also had substantial repeatability, although slightly less than the Obel method, (0.80 vs. 0.91). Excellent agreement between the methods was found, with the mean difference (95% CI), comparing the Obel grade, with the "modified Obel" score converted to an Obel grade, being -0.12 (-0.19 to -0.06) grades. The Obel and converted "modified Obel" grades were identical 62% of the time (259/420) and a difference of one grade (higher or lower) occurred in 35% of cases (148/420).Conclusion: Both methods show excellent agreement, reproducibility and repeatability when used to diagnose endocrinopathic laminitis. The "modified Obel" method is a three-step examination process for severity-scoring of endocrinopathic laminitis, initially proposed for use within a research setting. When using the modified method a diagnosis of laminitis also requires clinical acumen. The allocation of scores for specific clinical signs should be particularly useful in research trials monitoring laminitis recovery

Carbohydrate pellets to assess insulin dysregulation in horses

Background: A glycemic challenge test is used for the diagnosis of insulin dysregulation (ID) in horses and ponies. Different forms of the test exist where the administrative route and dose of glucose vary, which makes interpretation of results challenging. Hypothesis/Objectives: To evaluate the palatability of, and blood glucose and insulin responses to, carbohydrate pellets fed as an oral glucose test (OGT), and to establish the diagnostic threshold for ID when using the pellets. Animals: University and privately-owned horses and ponies (n = 157) comprised of 31 breeds and both sexes. Methods: Multicenter cohort study. A custom-produced glycemic pellet was offered for free intake at 0.5 g/kg BW soluble carbohydrate and serum insulin and blood glucose concentrations measured before and after (60, 120, and 180 minutes) the pellets were offered. Pellet acceptance and intake time (those that finished within 10 minutes) were determined to assess palatability. Results: The pellets were palatable to 132/157 animals, and ponies found the pellets more (P =.004) palatable than horses. The median intake time (4 [3-6] minutes) was positively correlated with acceptance grade (r =.51; P <.0001). Consumption of the pellets elicited peak blood glucose (6.6 [5.8-7.8] mmol/L) and serum insulin (40.5 [19-99.8] μIU/mL) responses at 120 minutes. At 120 minutes the optimal cut-off was 83 μIU/mL (95% CI: 70-99 μIU/mL) for the IMMULITE 2000XPi assay. Conclusions and Clinical Importance: The pellets were palatable and a suitable, novel carbohydrate source for the OGT.</p

Carbohydrate pellets to assess insulin dysregulation in horses.

BackgroundA glycemic challenge test is used for the diagnosis of insulin dysregulation (ID) in horses and ponies. Different forms of the test exist where the administrative route and dose of glucose vary, which makes interpretation of results challenging.Hypothesis/objectivesTo evaluate the palatability of, and blood glucose and insulin responses to, carbohydrate pellets fed as an oral glucose test (OGT), and to establish the diagnostic threshold for ID when using the pellets.AnimalsUniversity and privately-owned horses and ponies (n = 157) comprised of 31 breeds and both sexes.MethodsMulticenter cohort study. A custom-produced glycemic pellet was offered for free intake at 0.5 g/kg BW soluble carbohydrate and serum insulin and blood glucose concentrations measured before and after (60, 120, and 180 minutes) the pellets were offered. Pellet acceptance and intake time (those that finished within 10 minutes) were determined to assess palatability.ResultsThe pellets were palatable to 132/157 animals, and ponies found the pellets more (P = .004) palatable than horses. The median intake time (4 [3-6] minutes) was positively correlated with acceptance grade (r = .51; P Conclusions and clinical importanceThe pellets were palatable and a suitable, novel carbohydrate source for the OGT

Glycine activates myenteric neurones in adult guinea-pigs

We studied the effects of glycine on myenteric neurones and muscle activity in the colon and stomach of adult guinea-pigs.Intracellular recordings revealed that myenteric neurones responded to local microejection of glycine (1 mm) with a fast, transient membrane potential depolarisation (57 % of 191 colonic neurones and 26 % of 50 gastric neurones). Most glycine-sensitive neurones had ascending projections and were choline acetyltransferase immunoreactive. Glycine preferentially activated neurones with a late afterhyperpolarisation (AH-neurones) and tonic spiking neurones with fast synaptic inputs (tonic S-neurones) but less frequently phasic S-neurones and inexcitable (non-spiking) neurones. The depolarisation had a reversal potential at −19 ± 13 mV, which was increased by 18 ± 10 % upon lowering extracellular chloride concentration and decreased by 38 ± 14 % in furosemide (frusemide, 2 mm).Strychnine (300 nm) reversibly abolished the glycine-induced depolarisation and the Cl− channel blocker picrotoxin (100 μm) reduced the amplitude of the depolarisation by 55 ± 5 %. The glycine effect was a postsynaptic response because it was not changed after nerve blockade with tetrodotoxin (1 μm) or blockade of synaptic transmission in reduced extracellular [Ca2+]. The effect was specific since the response was not changed by the nicotinic antagonists hexamethonium (200 μm) and mecamylamine (100 μm), the GABAA receptor antagonist bicuculline (10 μm), the NMDA antagonist MK-801 (20 μm) or the 5-HT3 antagonist ICS 205930 (1 μm).Glycine (1 mm) induced a tetrodotoxin- and strychnine-sensitive contractile response in the colon; the contractile response in the stomach was tetrodotoxin insensitive.Glycine activated myenteric neurones in the adult enteric nervous system through strychnine-sensitive mechanisms. The glycine-evoked depolarisation was caused by Cl− efflux and the maintenance of relatively high intracellular chloride concentrations involved furosemide-sensitive cation-chloride co-transporters