Insulin dysregulation (ID), core to equine metabolic syndrome, may present without obesity. Testing for ID risk is commonly based on breed and obese phenotype but might be valuable for non-obese stock-type horses. This study aimed to determine the prevalence of ID in non-obese stock-type horses and evaluate if morphometric neck measurements (MNM) correlate with ID. Sixty-two, non-obese (BCS 5, range 2.5-6/9) stock-type horses were assessed for MNM: neck circumference at 25%, 50% (NC50), and 75% (NC75) length, and crest height. An oral sugar test (OST; 0.15 mL/kg BW corn syrup) was performed with blood taken pre- and 60 min post-OST for insulin (PREI, POSTI) and glucose (PREG, POSTG). Insulin dysregulation was defined as insulin concentration > 45 µIU/mL POSTI. Three of 62 horses were ID (4.8%; 95% CI 1.0%-13.5%). Horses with ID had greater PREG (121.0 ± 7.56 vs. 105.3 ± 1.72 mg/dL; LS means ± SEM; P = 0.04) and PREI (15.7 ± 2.63 vs. 10.5 ± 0.59 µIU/mL; P = 0.05) than normal responders. Mares had greater PREI than geldings (11.7 ± 0.76 vs. 9.4 ± 0.89 µIU/mL; P = 0.04). Stepwise regression indicated a weak relationship with crest height and POSTG (y = 51.27 + (0.88 x NC50); R2 = 0.09; P = 0.02). Post-glucose correlated with NC50 (r = 0.30; P = 0.04) and NC75 (r = 0.29; P = 0.03). This study showed 4.8% of non-obese horses had ID, warranting testing irrespective of phenotype, but only a weak association between MNM and POSTG was found.

OBJECTIVE: Congenital hyperinsulinism (CHI) is a rare, monogenic disease characterized by excessive insulin secretion. We aimed to evaluate all probands with suspected CHI in Norway registered over the past two decades.
METHODS: The study included 98 probands. Clinical data were cumulated from medical records. All probands were screened for variants in the genes ABCC8 and KCNJ11. Other CHI-related genes were Sanger-sequenced as indicated by the patients\' phenotype (N=75) or analyzed by next-generation sequencing employing a panel of 30 CHI-related genes (N=23).
RESULTS: Twenty-one probands (21%) received a diagnosis other than CHI, the most common being idiopathic ketotic hypoglycemia (9%) or syndromic hyperinsulinism (4%). In the final cohort of 77 CHI probands, genetic findings were revealed in 46 (60%). ABCC8 variants were most common (N=40) and five novel variants were identified. One proband harbored both the pathogenic GCK variant p.(Ala456Val) and the ABCC8 variant p.(Gly505Cys). Although most ABCC8 variants caused immediate disease onset with severe hypoglycemia and were diazoxide-unresponsive, eight probands had a heterozygous, apparently dominant variant with milder phenotype. Two probands had pathogenic variants in GLUD1, whereas variants in HADH, HNF4A, KCNJ11, and HK1 were identified in one proband each, the latter being non-coding. Neurologic sequelae were reported in 53% of the CHI probands. Of non-surgically treated probands, 43% had spontaneous resolution. The minimum birth prevalence of CHI in Norway is 1:19,400 live births.
CONCLUSIONS: Individuals with disease-causing ABCC8 variants dominated our cohort. Patients with known genetic etiology had earlier and more severe disease-onset than genetically unsolved patients.

In insulin dysregulation, hyperinsulinemia (HI) can be accompanied by peripheral insulin resistance (IR) in horses, which can be diagnosed with an insulin-tolerance test (ITT). The administration of 0.1 IU/kg body weight of recombinant regular human insulin (RHI) should elicit a 50 % reduction of the initial blood glucose concentration at 30 min after insulin administration in insulin sensitive horses. Compared to RHI, porcine zinc insulin (PZI) is veterinary-approved and therefore easier accessible for many practitioners. The aim of this study was to compare the insulin and glucose dynamics during a standard ITT with RHI to an ITT performed with PZI. Twelve Icelandic horses were subjected to an ITT with RHI (ITT-RHI) and with PZI (ITT-PZI) at same dosages in a randomised crossover design. The insulin and glucose dynamics that resulted from these tests were compared, and the consistency of classification into insulin-sensitive and IR categories was evaluated. No complications were observed with the use of either RHI or PZI in ITT. A good correlation of the test results was observed (r = 0.88; P < 0.001). The blood glucose concentrations and the percentage reduction in glucose concentration did not differ significantly between the two tests (P = 0.053), but four out of twelve horses were classified as IR in the ITT-RHI whereas with the ITT-PZI seven out of twelve horses were classified as IR with the 50 % glucose reduction from baseline. Based on the Youden index, when using the ITT-PZI, an adjusted cut-off value for blood glucose reduction of 40 % at 30 min resulted in better test performance. With consideration for the seemingly weaker effect of PZI and the adjusted cut-off value, PZI can be an appropriate substitute to RHI in an ITT.

Obesity is a complex disease associated with augmented risk of metabolic disorder development and cellular dysfunction in various species. The goal of the present study was to investigate the impacts of obesity on the metabolic health of old mares as well as test the ability of diet supplementation with either a complex blend of nutrients designed to improve equine metabolism and gastrointestinal health or L-carnitine alone to mitigate negative effects of obesity. Mares (n = 19, 17.9 ± 3.7 years) were placed into one of three group: normal-weight (NW, n = 6), obese (OB, n = 7) or obese fed a complex diet supplement for 12 weeks (OBD, n = 6). After 12 weeks and completion of sample collections, OB mares received L-carnitine alone for an additional 6 weeks. Obesity in mares was significantly associated with insulin dysregulation, reduced muscle mitochondrial function, and decreased skeletal muscle oxidative capacity with greater ROS production when compared to NW. Obese mares fed the complex diet supplement had better insulin sensivity, greater cell lipid metabolism, and higher muscle oxidative capacity with reduced ROS production than OB. L-carnitine supplementation alone did not significantly alter insulin signaling, but improved lipid metabolism and muscle oxidative capacity with reduced ROS. In conclusion, obesity is associated with insulin dysregulation and altered skeletal muscle metabolism in older mares. However, dietary interventions are an effective strategy to improve metabolic status and skeletal muscle mitochondrial function in older mares.

The aim of this study was to determine the effect of body condition score (BCS) on metabolic and endocrine parameters in pregnant Criollo mares (n=41), which were categorized according to their BCS as obese (7 to 9 BCS, n=26) or normal (5 to 7, n=15). Blood samples were taken during gestation in 3 periods: between 3.5 and 5 months (I), 8 and 9 months (II) and in the last month of gestation (III). The data was analyzed in the statistical model by mixed procedures, including BCS, gestational period and their interaction as fixed effects. BCS was only different in period I, as normal mares increased their BCS in the later periods. Leptin concentrations were greater in obese mares when compared to non-obese mares during all sampling periods (P<0.01), while glucose concentrations were also greater in the former group (P<0.01) but only during the first sampling period. Insulin concentrations and homeostatic model assessment for insulin resistance (HOMA-IR) index were greater also in obese mares in periods I and III (P<0.05). Adiponectin concentrations in period I were lower in obese mares (P<0.05). Cholesterol concentrations increased during gestation, and obese mares tended to have greater concentrations than nonobese mares (P<0.1). Triglyceride concentrations were not affected by group or gestational period. This study revealed adaptations in carbohydrate and lipid metabolism during gestation in mares. Several parameters are dependent on the degree of body fat reserves, which are reflected in the concentrations of biomarkers such as leptin and adiponectin. Insulin concentration in obese mares was higher than non-obese mares at the end of gestation, a similar profile was observed for HOMA-IR although cutoff values are yet still to be validated.

Glucagon-like peptide-1 (GLP-1), the principal incretin in horses, may play a role in the pathophysiology of insulin dysregulation (ID). This study aimed to describe its concentration in response to three preserved forages and four dynamic tests for ID in ponies. Twelve adult ponies of mixed ID status were given a meal of hay, soaked hay or haylage, an in-feed oral glucose test (OGT), oral sugar test (OST), an oral test using a proprietary breakfast cereal (WEET) or a combined glucose-insulin tolerance test (CGIT) weekly in a randomised cross-over study. Glucose, insulin and GLP-1 concentrations were measured before and following each intervention. Ponies were designated ID or non-ID and insulin resistant (IR) or non-IR according to OGT and CGIT results, respectively. All interventions apart from the CGIT provoked a GLP-1 response within 30 min. The OGT and WEET interventions, (containing the greatest dose of non-structural carbohydrate, 1.06 and 1 g/kg BW, respectively), resulted in a greater area under the curve (AUC) for GLP-1 compared to all other interventions (P < 0.001). No difference in GLP-1 response was detected according to ID or IR status, despite there being strong positive correlations (rs [95 % CI]) between GLP-1 and insulin concentrations measured at individual time points (0.67 [0.62 - 0.71]; P < 0.001) and as AUC (0.66 [0.49-0.79], P < 0.001). These data do not support of the use of GLP-1 as an adjunctive diagnostic test for ID or IR, as defined by conventional intravenous or oral dynamic tests.

Obesity is an important health concern in horses, along with humans and companion animals. Adipose tissue is an inflammatory organ that alters the insulin-signaling cascade, ultimately causing insulin dysregulation and impaired glucose metabolism. These disruptions can increase the risk of metabolic disease and laminitis in horses and may also impact energy metabolism during exercise. A single bout of exercise, along with chronic exercise conditioning, increases insulin sensitivity and glucose disposal via both contraction- and insulin-mediated glucose uptake pathways. Regular exercise also increases calorie expenditure, which can facilitate weight (as body fat) loss. This paper explores the metabolic pathways affected by adiposity, as well as discusses the impact of exercise on insulin metabolism in horses.

OBJECTIVE: To evaluate the feasibility of using the FreeStyle Libre (a continuous glucose monitoring system [CGMS]) for instantaneous continuous monitoring of interstitial glucose in adult horses and examine the applicability and accuracy of this system in horses submitted to combined glucose-insulin test (CGIT).
METHODS: Laboratory measurements and continuous glucose monitoring system (CGMS) readings were analyzed using a 2 × 2 factorial statistical model with repeated measures over time. This analysis assessed the effects of the test (factor 1), group (factor 2), and their interactions (test × group, test × time, and group × time). Pearson\'s correlation analysis was applied to blood glucose values. Mean comparisons were conducted using the t-test, and agreement between techniques was assessed via the Bland-Altman method, with a 95% confidence interval.
METHODS: Field study on private horse farms in association with a veterinary school.
METHODS: Ten healthy stallions were assigned to one of two groups based on their body condition scores (BCS). Group 1 (G1, n = 5) consisted of nonobese horses with a BCS of 5 or 6, while Group 2 (G2, n = 5) consisted of obese horses with a BCS of 7 or higher.
METHODS: A CGMS sensor was attached to the dorsolateral aspect of the proximal one third of each horse\'s neck. Laboratory blood glucose measurements and CGMS interstitial glucose readings were compared at different time points for up to 7 days after sensor fixation. Obese horses were also submitted to CGIT on Day 4.
RESULTS: A comparative analysis of glucose measurements obtained in G1 and G2 horses using the CGMS and enzymatic methods revealed significant group × time interactions (P < 0.001) and time effects (P < 0.001). No interactions were detected between group (P = 0.45), test (P = 0.62), group and test (P = 0.28), or time and test (P = 0.92). In G1 and G2, tests were significantly correlated (r = 0.84 and P = 0.00) at all time points (T0-T5). Agreement between the glucose values obtained using different methods was excellent despite a small time delay in CGMS detection of rapid changes in blood glucose.
CONCLUSIONS: It was concluded that the CGMS can be used for indirect assessment of glycemic status (ie, based on interstitial glucose measurements) in nonobese and obese adult horses submitted to CGIT.

Equine metabolic syndrome (EMS) is a common welfare problem in horses worldwide. It is characterized by insulin dysregulation (ID), predisposition to laminitis and often obesity. EMS is multifactorial by nature, with both the environment and genetics contributing to the phenotype. Environmental factors, such as feeding and exercise, can be controlled, thus forming the basis for treatment and prevention. Genetic factors, by contrast, are less well-known and not easily controllable. The aim of this study was to identify potential genetic loci influencing ID/EMS in Finnhorses. A single-breed (Finnhorse) case-control genome-wide association study (GWAS) of ID was conducted with controls that included age-appropriate non-ID horses. ID status was determined with an oral sugar test (OST) for fasted horses. Seventy-one Finnhorses participated (n = 34 ID, n = 37 control). DNA samples (hair roots) were genotyped for 65 157 single-nucleotide polymorphisms (SNPs) with the Illumina Equine SNP70 BeadChip, and these data were analysed for association and FST outliers with genomic tools. P-values that exceeded the suggestive threshold (P = 1.00 ×10-5) were found in SNP BIEC2_383954 (P = 3.45 ×10-6) in chromosome 17 and SNP BIEC2_312374 (P = 1.89 ×10-5) in chromosome 15. Hierarchical and Bayesian FST outlier tests also detected these SNPs. Potential candidate genes associated with the ID close to SNP BIEC2_383954, with functions in carbohydrate metabolism, were Arginine and Glutamate Rich 1 (ARGLU1) and Ephrin-B2 (EFNB2).

Hyperinsulinemia-associated laminitis (HAL) is the primary concern for insulin dysregulated (ID) equids and their insulin response to the consumption of oral, nonstructural carbohydrates (NSC) has been shown to be a risk predictor for HAL development. This randomized, crossover study\'s objective was to examine the insulinemic responses to 3 forage pellets (1 g/kg BW) (timothy hay, TH, 9.5 % CP & 10 % NSC DM; alfalfa hay, AH, 16.3 % CP & 9.8 % NSC DM; timothy-alfalfa hay; TAH, 17.2 % CP & 9.8 % NSC DM) along with a positive (dehulled oats; OG, 14.7 % CP & 59.7 % NSC DM) and negative dietary challenge control (low-NSC; LNSC, 12.8 % CP & 5.4 % NSC DM) of ID (n = 8; 16.1 ± 2.2 yr; 565.4 ± 99.1 kgs.) and non-ID (NID; n = 7; 17.0 ± 2.8 yr; 583.6 ± 57.9 kgs.) horses. ID horses had higher positive incremental area under the curve for insulin (IAUCi) (ID: 890 ± 925 µIU/mL*minute vs. NID: 225 ± 228 µIU/mL*minute), peak (ID: 101.5 ± 80.72 µIU/mL vs. NID: 25.7 ± 7.2 µIU/mL), and delta (ID: 45.5 ± 77.1 µIU/mL vs. NID: 4.9 ± 5.3 µIU/mL) insulin for all forage pellets compared to NID (p < 0.01). ID horses IAUCi for the forage pellets was not different compared to the LNSC (218 ± 327 µIU/mL*minute) but was different from OG (10,522 ± 4,565 µIU/mL*minute). ID horses\' lack of an augmented insulinemic response to the low NSC forage pellets (fed in small amounts) indicates that they could be a safe feedstuff for ID animals.