Factors associated with insulin responses to oral sugars in a mixed-breed cohort of ponies.

BACKGROUND: Serum insulin concentration at 60 min (InsulinT60) during an oral sugar test (OST) indicates future laminitis risk and insulin dysregulation (ID). Associations between InsulinT60 and physical and owner-reported variables may help clinicians select individuals for ID testing. Associations between InsulinT60 and other metabolic markers may help elucidate ID pathophysiology. OBJECTIVES: To describe associations between (A) season, physically-apparent and owner-reported factors and binary InsulinT60 interpretation (initial models) and (B) variables included in the initial models, other metabolic markers and continuous InsulinT60 (full models). STUDY DESIGN: Prospective longitudinal. METHODS: Non-laminitic ponies were examined and OSTs (0.3 mL/kg Karo syrup) performed every 6 months (autumn and spring) for ≤4 years. Factors associated with InsulinT60 were determined using mixed-effects models with physical, owner-reported, season and serum/plasma markers as fixed effects and pony and premises identifiers as random effects. Autumn and spring data were analysed separately for full models. RESULTS: One thousand seven hundred and sixty-three OSTs from 367 ponies were included. High-risk InsulinT60 (>153 µIU/mL) was independently associated with (odds ratio, 95% confidence interval [CI]): age (1.36, 1.16-1.60), body condition score (BCS) (2.38, 1.21-4.69), and bulging supraorbital fatpads (7.25, 2.1-24.98). However, the initial models provided little explanatory power (Nakagawa R2 = 0.1-0.27). LoginsulinT60 was independently associated with (effect estimate, 95% CI): age (0.02, 0.01-0.04), Welsh/Welsh X breed (0.22, 0.05, 0.39), sex (gelding = -0.2, -0.34 to 0.06), BCS (0.16, 0.08-0.23), plasma adiponectin (-0.02, -0.02 to 0.01) and basal insulin (0.01, 0.01-0.01) in spring, and: age (0.03, 0.02-0.04), BCS (0.17, 0.08-0.26), bulging supraorbital fatpads (0.37, 0.2-0.54), turnout score (0.05, 0.02-0.09), plasma adiponectin (-0.01, -0.02 to 0.01), ACTH (per 10 pg/mL) (0.01, 0.00-0.01), triglycerides (0.28, 0.07-0.49) and InsulinT0 (0.01, 0.01-0.01) in autumn. MAIN LIMITATIONS: Only non-laminitic ponies in one region were included. CONCLUSIONS: Owner-reported and physically-apparent data were limited InsulinT60 predictors. InsulinT60 is a complex trait, independently associated with numerous variables, some with seasonal interactions.

Insulin, but not adiponectin, is detectable in equine saliva using an automated, commercial assay.

BACKGROUND: The hormones insulin and adiponectin are commonly measured in equids because of their involvement in endocrinopathic laminitis. These are currently measured in serum/plasma, although jugular venipuncture can cause stress in some animals and may be impossible in needle-shy individuals. However, both hormones can be measured in saliva in other species. OBJECTIVES: To determine whether [insulin] and [total adiponectin] are detectable in equine saliva using automated assays and whether saliva collection is associated with changes in stress indicators. Additionally, the correlation between serum and salivary [insulin] was investigated. STUDY DESIGN: In vivo experiment. METHODS: Paired blood and saliva samples were collected from eight adult ponies at multiple time-points (n = 45 paired samples). [Insulin] and [total adiponectin] were measured using automated assays validated for equine serum/plasma. Blink rates and heart rates were determined, using video recordings and a wearable heart rate monitor respectively, to assess the effects of sample collection on stress indicators compared with a control situation without a stressful stimulus. RESULTS: [Total adiponectin] was undetectable in saliva. However, salivary [insulin] was measurable with acceptable inter-assay (1.3 ± 0.9%) and intra-assay (1.1 ± 0.6%) variability. Blink and heart rates during saliva collection did not differ significantly from those in the control setting. Serum and salivary [insulin] were not significantly correlated. MAIN LIMITATIONS: Small sample size comprising native UK ponies; potential sampling bias as animals were recruited to the study partly based on their behaviour during blood sampling; saliva collected from unfasted animals. CONCLUSIONS: Insulin is measurable in equine saliva using an automated assay currently available in the UK, but further validation and the determination of specific diagnostic thresholds are required. Saliva collection was not associated with changes in stress indicators. Further research is therefore needed to determine the potential of equine saliva as a non-invasive alternative to blood for insulin determination.

BEVA primary care clinical guidelines: Diagnosis and management of equine pituitary pars intermedia dysfunction.

BACKGROUND: Pituitary pars intermedia dysfunction (PPID) is a prevalent, age-related chronic disorder in equids. Diagnosis of PPID can be challenging because of its broad spectrum of clinical presentations and disparate published diagnostic criteria, and there are limited available treatment options. OBJECTIVES: To develop evidence-based primary care guidelines for the diagnosis and treatment of equine PPID based on the available literature. STUDY DESIGN: Evidence-based clinical guideline using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) framework. METHODS: Research questions were proposed by a panel of veterinarians and developed into PICO or another structured format. VetSRev and Veterinary Evidence were searched for evidence summaries, and systematic searches of the NCBI PubMed and CAB Direct databases were conducted using keyword searches in July 2022 and updated in January 2023. The evidence was evaluated using the GRADE framework. RESULTS AND RECOMMENDATIONS: The research questions were categorised into four areas: (A) Case selection for diagnostic testing, pre-test probability and diagnostic test accuracy, (B) interpretation of test results, (C) pharmacological treatments and other treatment/management options and (D) monitoring treated cases. Relevant veterinary publications were identified and assessed using the GRADE criteria. The results were developed into recommendations: (A) Case selection for diagnostic testing and diagnostic test accuracy: (i) The prevalence of PPID in equids aged ≥15 years is between 21% and 27%; (ii) hypertrichosis or delayed/incomplete hair coat shedding provides a high index of clinical suspicion for PPID; (iii) the combination of clinical signs and age informs the index of clinical suspicion prior to diagnostic testing; (iv) estimated pre-test probability of PPID should be considered in interpretation of diagnostic test results; (v) pre-test probability of PPID is low in equids aged <10 years; (vi) both pre-test probability of disease and season of testing have strong influence on the ability to diagnose PPID using basal adrenocorticotropic hormone (ACTH) or ACTH after thyrotropin-releasing hormone (TRH) stimulation. The overall diagnostic accuracy of basal ACTH concentrations for diagnosing PPID ranged between 88% and 92% in the autumn and 70% and 86% in the non-autumn, depending on the pre-test probability. Based on a single study, the overall diagnostic accuracy of ACTH concentrations in response to TRH after 30 minutes for diagnosing PPID ranged between 92% and 98% in the autumn and 90% and 94% in the non-autumn, depending on the pre-test probability. Thus, it should be remembered that the risk of a false positive result increases in situations where there is a low pre-test probability, which could mean that treatment is initiated for PPID without checking for a more likely alternative diagnosis. This could compromise horse welfare due to the commencement of lifelong therapy and/or failing to identify and treat an alternative potentially life-threatening condition. (B) Interpretation of diagnostic tests: (i) There is a significant effect of breed on plasma ACTH concentration, particularly in the autumn with markedly higher ACTH concentrations in some but not all 'thrifty' breeds; (ii) basal and/or post-TRH ACTH concentrations may also be affected by latitude/location, diet/feeding, coat colour, critical illness and trailer transport; (iii) mild pain is unlikely to have a large effect on basal ACTH, but caution may be required for more severe pain; (iv) determining diagnostic thresholds that allow for all possible contributory factors is not practical; therefore, the use of equivocal ranges is supported; (v) dynamic insulin testing and TRH stimulation testing may be combined, but TRH stimulation testing should not immediately follow an oral sugar test; (vi) equids with PPID and hyperinsulinaemia appear to be at higher risk of laminitis, but ACTH is not an independent predictor of laminitis risk. (C) Pharmacologic treatments and other treatment/management options: (i) Pergolide improves most clinical signs associated with PPID in the majority of affected animals; (ii) Pergolide treatment lowers basal ACTH concentrations and improves the ACTH response to TRH in many animals, but measures of insulin dysregulation (ID) are not altered in most cases; (iii) chasteberry has no effect on ACTH concentrations and there is no benefit to adding chasteberry to pergolide therapy; (iv) combination of cyproheptadine with pergolide is not superior to pergolide alone; (v) there is no evidence that pergolide has adverse cardiac effects in horses; (vi) Pergolide does not affect insulin sensitivity. (D) Monitoring pergolide-treated cases: (i) Hormone assays provide a crude indication of pituitary control in response to pergolide therapy, however it is unknown whether monitoring of ACTH concentrations and titrating of pergolide doses accordingly is associated with improved endocrinological or clinical outcome; (ii) it is unknown whether monitoring the ACTH response to TRH or clinical signs is associated with an improved outcome; (iii) there is very weak evidence to suggest that increasing pergolide dose in autumn months may be beneficial; (iv) there is little advantage in waiting for more than a month to perform follow-up endocrine testing following initiation of pergolide therapy; there may be merit in performing repeat tests sooner; (v) timing of sampling in relation to pergolide dosing does not confound measurement of ACTH concentration; (vi) there is no evidence that making changes after interpretation of ACTH concentrations measured at certain times of the year is associated with improved outcomes; (vii) evidence is very limited, however, compliance with PPID treatment appears to be poor and it is unclear whether this influences clinical outcome; (viii) evidence is very limited, but horses with clinical signs of PPID are likely to shed more nematode eggs than horses without clinical signs of PPID; it is unclear whether this results in an increased risk of parasitic disease or whether there is a need for more frequent assessment of faecal worm egg counts. MAIN LIMITATIONS: Limited relevant publications in the veterinary scientific literature. CONCLUSIONS: These findings should be used to inform decision-making in equine primary care practice.

Short-term induced hyperinsulinaemia and dexamethasone challenge do not affect circulating total adiponectin concentrations in insulin-sensitive ponies.

BACKGROUND: Hypoadiponectinaemia is a risk factor for endocrinopathic laminitis, but the directionality and nature of its association with insulin dysregulation is unclear. OBJECTIVES: To investigate the effects of short-term induced hyperinsulinaemia and dexamethasone challenge on circulating [total adiponectin] and whole blood expression of adiponectin (AdipoR1 and AdipoR2), insulin, and insulin-like growth factor 1 (IGF-1) receptors in insulin-sensitive ponies. STUDY DESIGN: In vivo experiment. METHODS: Six never-laminitic, insulin-sensitive, native-breed UK ponies first underwent a dexamethasone challenge (0.08 mg/kg i.v.) with blood samples collected every 15 min over 3 h. After a 14-day washout period, hyperinsulinaemia was induced for 9 h via a euglycaemic-hyperinsulinaemic clamp (EHC), with blood samples collected every 30 min. Serum [insulin], plasma [total adiponectin], and plasma [IGF-1] were measured using validated assays and receptor gene expression was assessed via quantitative polymerase chain reaction (qPCR). Finally, whole blood was incubated with 10-1000 ng/mL dexamethasone for 3 h at 37°C to investigate its direct effects on gene expression. RESULTS: There were no adverse effects observed during either protocol. Dexamethasone challenge did not alter circulating [insulin] or [total adiponectin] at any time-point, but significantly upregulated AdipoR1 and IGF-1R expression at 150 and 180 min. Ex vivo incubation of whole blood with dexamethasone did not alter expression of the genes examined. There was no change in [total adiponectin] or expression of the genes examined associated with EHC-induced hyperinsulinemia. MAIN LIMITATIONS: This was a small sample size that included only native-breed ponies; total adiponectin was measured rather than high-molecular-weight adiponectin. CONCLUSIONS: Short-term induced hyperinsulinaemia and dexamethasone challenge did not affect circulating [total adiponectin] in insulin-sensitive ponies. However, dexamethasone administration was associated with upregulation of two receptors linked to adiponectin signalling, suggesting that a physiological response occurred possibly to counteract dexamethasone-associated changes in tissue insulin sensitivity.

Relationships between total adiponectin concentrations and obesity in native-breed ponies in England.

BACKGROUND: Equine metabolic syndrome is a collection of risk factors associated with an increased risk of endocrinopathic laminitis. All affected animals display insulin dysregulation and some may show adiponectin dysregulation and/or excessive adiposity. However, the relationship between obesity and hypoadiponectinaemia in equids remains unclear. OBJECTIVES: To investigate the relationship between obesity and circulating plasma total adiponectin (TA) concentrations in native-breed ponies in England. STUDY DESIGN: Secondary data analysis. METHODS: Data collected for three previous studies were retrospectively analysed and cohorts were pooled where possible (maximum sample size: n = 734 ponies). Correlations between [TA], age, and morphometric measures were assessed using Spearman's correlation coefficient. [TA] was compared between animals of different body condition score (BCS) classification (ideal-weight, overweight, and obese), breed, and body shape using Kruskal-Wallis with Dunn's post hoc tests, and sex using Mann-Whitney U test. The proportions of obese and ideal-weight ponies with basal hyperinsulinaemia and/or hypoadiponectinaemia were compared using a Chi-square test of homogeneity and post hoc z-test. Logistic regression was used to identify factors that may discriminate ponies with hypoadiponectinaemia. RESULTS: [TA] was weakly positively correlated with BCS, height, weight, and weight:height ratio (Spearman's ρ = 0.14-0.29, p < 0.05). There were significant differences in [TA] in ponies with different BCS group classification, body shape, and breed. A greater percentage of obese (54.6%) than ideal-weight ponies (33.1%, p < 0.001) had both normal [TA] and [basal insulin], and a greater percentage of ideal-weight (38.6%) than obese ponies (16.5%, p < 0.001) showed hypoadiponectinaemia. Weight:height and BCS group were significant variables in a logistic regression of hypoadiponectinaemia but model fit and predictive accuracy were poor. MAIN LIMITATIONS: Retrospective study design, only native-breed ponies included. CONCLUSIONS: Morphometric measures such as BCS do not closely reflect [TA]. Circulating [TA] and [basal insulin] should be determined in all animals with predisposing factors, regardless of obesity status.

Association between insulin dysregulation and adrenocorticotropic hormone in aged horses and ponies with no clinical signs of pituitary pars intermedia dysfunction.

BACKGROUND: High concentrations of adrenocorticotropic hormone (ACTH) are used as an indicator of pituitary pars intermedia dysfunction (PPID), but other factors that may influence ACTH need to be understood, if diagnostic reference ranges for ACTH are to be used with confidence. Insulin dysregulation (ID) could be one such factor, as insulin affects pituitary hormones in other species. OBJECTIVES: To test the hypothesis that a relationship exists between high insulin and high ACTH in aged (>15-year-old) animals with no clinical signs of PPID. STUDY DESIGN: A cohort study. METHODS: Thirteen horses and eleven ponies (17-25 years-old; mares and geldings) were clinically examined for signs of PPID in the spring (November 2020) and autumn (April 2021). On the same day, blood samples were taken before and 2 h after an oral glucose test (OGT). Concentrations of insulin, glucose, ACTH and cortisol were measured. RESULTS: There was no association between ACTH and cortisol. However, there was a positive linear correlation between ACTH and post-OGT (insulin in the autumn (r = 0.427, p = 0.04). Two horses and six ponies had ACTH above the cut-off value for PPID diagnosis, and of these eight animals, six also had insulin concentrations above the cut-off value for ID. MAIN LIMITATIONS: The cohort was small and thyrotropin-releasing hormone (TRH) stimulation tests were not performed. CONCLUSIONS: In autumn, high ACTH was associated with ID, when no clinical signs of PPID were present. Because ACTH is used in PPID diagnosis, further work is required to understand this interaction.

Predictors of laminitis development in a cohort of nonlaminitic ponies.

BACKGROUND: Quantifying risk factors for laminitis development requires improvement. OBJECTIVES: To identify the most useful physical examination, metabolic and management factors to predict laminitis development in client-owned, nonlaminitic ponies. STUDY DESIGN: Prospective cohort study. METHODS: Physical examination, metabolic and management data were collected from a pony cohort 6 monthly for up to 4 years. Ponies were monitored for the development of laminitis. Metabolic data included basal plasma concentrations of ACTH ([ACTH]), adiponectin ([adiponectin]), triglycerides and glucose. Serum insulin concentrations ([insulin]) were measured in the unfasted basal state ([insulin]T0) and 60 minutes ([insulin]T60) after administration of corn syrup (0.3ml/kg). Separate multivariable Cox proportional-hazards models were developed for physical, management/signalment and metabolic data and later combined into two final models. Low-, medium- and high-laminitis risk categories were defined based on basal or T60 [insulin]. RESULTS: Overall, 374 ponies (age 5-32 years) and 891 pony-years were included in the main analysis. Laminitis incidence (95% confidence interval (CI)) was 4.8 (3.5-6.5) cases/100 pony-years. Laminitis development was associated with numerous univariable factors. Significant (P < .05) factors retained in the final multivariable models included [insulin]T0, [insulin]T60, [adiponectin] and divergent hoof growth. [ACTH] was not independently associated with laminitis. Based on [Insulin]T0, low- (<21.6 µIU/ml), medium- (21.6-45.2 µIU/ml) and high-risk (>45.2 µIU/ml) categories encompassed 70, 20 and 10% of the population and had estimated 4-year laminitis incidences (95%CI) of 6 (2-9)%, 22 (10-33)% and 69 (48-82)% respectively. Based on [Insulin]T60 the low- (<53.4 µIU/ml), medium- (53.4-153 µIU/ml) and high-risk (≥153 µIU/ml) categories comprised 60, 30 and 10% of the population and had estimated 4-year laminitis incidences (95%CI) of 3 (0-6)%, 20 (10-29)% and 73 (52-84)% respectively. MAIN LIMITATIONS: Results may not apply to different insulin assays, geographical regions, breeds or management types. CONCLUSIONS: [Insulin]T0 or [insulin]T60 best quantify the risk of future laminitis development in nonlaminitic ponies.

Outcome following emergency laparotomy in 33 UK donkeys: A retrospective multicentre study.

BACKGROUND: Emergency laparotomies in donkeys are infrequently performed and there is limited literature on the subject. OBJECTIVES: To determine findings and associated outcomes of exploratory laparotomies in donkeys. STUDY DESIGN: Descriptive retrospective study. METHODS: Donkeys undergoing emergency exploratory laparotomy for investigation and treatment of colic at seven UK referral hospitals between 2005-2017 were included. Data were retrieved from available hospital records. Descriptive statistics and inferential statistical analysis of outcomes of interest was performed in three steps. RESULTS: Thirty-three cases fulfilled the inclusion criteria. Clinical signs on presentation were available for 32 donkeys, of which 53.1% (17/32) presented for investigation of colic while in 46.9% (15/32) the presenting complaint was non-specific. Primary lesion location included small intestine (42.4%, 14/33), large colon (39.3%, 13/33), caecum (6.1%, 2/33), stomach (6.1%, 2/33) and 6.1% (2/33) had multiple abnormal findings without a clear primary lesion. Overall survival to discharge was 54.5% (18/33). Five donkeys (15.2%, 5/33) were euthanased at surgery and of those recovering from general anaesthesia a further 35.7% (10/28) were euthanased or died prior to discharge. Six donkeys (21.4%, 6/28) required a second laparotomy of which 4 (66.7%, 4/6) survived. Post-operative complications occurred in 82.1% (23/28) of cases and included hyperlipaemia (42.9%, 12/28), incisional complications (21.4%, 6/28), ileus (21.4%, 6/28) and persistent colic (17.9%, 5/28). When adjusted for other complications, donkeys with primary gastric lesions were less likely to have presented with severe colic compared with those with primary small intestinal lesions (OR: 0.07, 95% CI 0.01-0.95, p = 0.05). Only age was positively associated with death prior to discharge (OR: 1.18, 95% CI 1.03-1.36, p = 0.02). MAIN LIMITATIONS: Small sample size and retrospective design. CONCLUSION: Donkeys with abdominal lesions may present with a range of signs often not including colic. Surgical findings were diverse and survival to discharge appears to be lower than in horses.

Nitric oxide synthase inhibition reveals differences in the nitric oxide pathway in previously laminitic ponies.

Previously laminitic (PL) ponies are reported to have higher blood pressure than non laminitic (NL) ponies. This relative hypertension may be related to endothelial cell dysfunction, similar to humans with metabolic syndrome. To investigate the relationship between laminitis predisposition and endothelial dysfunction, the effect of nitric oxide synthase (NOS) inhibition on the change in circulating nitric oxide (NO) concentrations and systemic blood pressure (BP) was determined. An intravenous NO sensor was used to measure changes in blood NO concentration during and after intravenous infusion of the NOS inhibitor L-nitroarginine methyl ester (L-NAME; 4 mg/kg IV) in PL and NL ponies. NO concentrations decreased and BP increased in response to L-NAME infusion, with a significantly (p = 0.02) greater rate of increase in systolic BP in PL (14.62 ± 1.88 mmHg/h) compared to NL (8.54 ± 1.88 mmHg/h) ponies. This greater effect on BP in PL compared to NL ponies is consistent with higher basal NO production in PL ponies, suggesting that the relative hypertension previously seen in PL ponies results in increased basal NO production, rather than being caused by reduced NO production as hypothesised. Up-regulation of the NO system may be a compensatory mechanism stimulated by the higher resting BP in PL ponies. Further investigation of the mechanism underlying the relative hypertension seen in PL ponies is required.

The Effect of Strip Grazing on Physical Activity and Behavior in Ponies.

This work aimed to determine the effect of strip grazing on physical activity in ponies using behavioral observations alongside accelerometers positioned at the poll. In study one, ten British native breed ponies were randomly assigned to paddock A (50 × 110 m) or B (50 × 110 m divided into seven equal strips with access to one additional strip per day) for seven days (n = 5/paddock). In study two, ten different British native breed ponies were randomly assigned for 14 days individually to (1) a control field where the animal was allowed complete access to their allotted area (n = 4); (2) a field that increased in size daily by moving a lead fence (n = 2); and (3) a field that was strip grazed using lead and back fences moved the same distance daily (n = 4). Accelerometer data were sorted into twenty-four-hour periods; each 10-second epoch was categorized as standing, grazing or locomoting using previously validated cut-off points; and time spent in each category for each day calculated. Behavioral monitoring was undertaken by direct observation on days 12-14 (study two only). Accelerometer and behavioral data were compared between grazing methods within each study. Strip grazing had no significant effect on the time spent in each physical activity category in either study. Behavioral observation revealed all ponies spent most time grazing ≤4 hours after fence moving and strip grazed ponies spent significantly more time grazing the newly available grass than elsewhere. Thus, strip grazing did not alter physical activity in ponies, but did result in preferential grazing of new grass.

Cross-sectional study to identify the prevalence of and factors associated with laminitis in UK donkeys.

BACKGROUND: Laminitis causes lameness in donkeys, but its prevalence and factors associated with disease remain uncertain. OBJECTIVES: To determine the prevalence of and identify factors associated with laminitis in donkeys. STUDY DESIGN: Retrospective cross-sectional study. METHODS: All donkeys at the Donkey Sanctuary, UK, October 2015 to March 2019 were included. For animals that had laminitis during this period, age, sex, weight, body condition score, and the onset date and type of each episode (first or recurrent, acute or chronic) were recorded. Additionally, management data, foot lesion score, endocrine data, other medical conditions, occurrence of foot trimming, surgical procedures, diagnostic imaging, behavioural modification therapy or movement between farms within the month prior were noted. Controls were animals that did not experience laminitis during this period and similar data were recorded. Multivariable logistic regression modelling assessed the differences between the control group and laminitis outcome groups (first, all laminitis, acute and chronic episodes). RESULTS: Altogether, 707 animals were included; 364 were control animals; 343 had a first episode of laminitis during the study period, of which 200/343 had no further episodes and 143/343 had recurrent episodes resulting in a total of 512 laminitis episodes and the period prevalence was 48.5% over 42 months. Overall, 180/512 (35%) laminitis episodes were acute and 332/512 (65%) were chronic. Compared with control animals, the laminitic outcome groups were significantly (P < .05) more likely to be younger (first episode), less likely to get extra feed (all four groups) or have an additional medical problem (first episode), and less likely to have undergone dental work, movement, imaging (all four groups) or surgery (first; all laminitis, chronic episodes) in the month preceding the episode. MAIN LIMITATIONS: These results may not be applicable to the wider donkey population. CONCLUSIONS: Laminitis commonly affects donkeys, but factors associated with donkey laminitis differ from those reported in horses.

Influence of endocrine disease on l-lactate concentrations in blood of ponies.

BACKGROUND: Blood l-lactate concentrations are higher in people with developing or established diabetes mellitus and insulin resistance. OBJECTIVES: To investigate whether blood l-lactate concentrations are positively correlated with measures of insulin dysregulation (ID) or increased autumnal ACTH concentrations in ponies. ANIMALS: Systemically healthy client-owned ponies (n = 101). METHODS: Prospective case-control study. Blood samples were obtained from 101 clinically healthy ponies. Breed, weight, height, and subjective and objective measures of body condition were recorded. Blood l-lactate, glucose, triglyceride, total adiponectin, and ACTH concentrations were measured and an oral sugar test (OST) was carried out. Correlations between blood l-lactate and variables of endocrine health were determined. RESULTS: Using a seasonal cutoff point of ACTH concentrations ≥47 pg/mL, 55 ponies had increased autumnal ACTH concentrations and 45 did not. Using a basal insulin concentration of >50 µiU/mL, 42 ponies were diagnosed with ID and 58 were not. Using a 60 minutes after OST cutoff point of >45 µiU/mL, 57 ponies had ID and 37 did not. Blood l-lactate concentrations were significantly lower in obese (average body condition score ≥ 7/9) compared to nonobese ponies (0.6 mmol/L; range, 0.0-1.9 mmol/L vs 0.8 mmol/L; range, 0.3-2.7 mmol/L; P = .01). No other significant correlations were detected. No differences were detected between ponies with and without increased autumnal ACTH concentrations (0.7 mmol/L; range, 0.0-2.7 mmol/L vs 0.7 mmol/L; range, 0.3-1.8 mmol/L; P = .84) and with and without ID (0.7 mmol/L; range, 0.3-2.7 mmol/L vs 0.8 mmol/L; range, 0.0-1.6 mmol/L; P = .63). CONCLUSIONS AND CLINICAL IMPORTANCE: Results do not support an effect of endocrine status on l-lactate concentrations in blood of ponies.

Pergolide dosing compliance and factors affecting the laboratory control of equine pituitary pars intermedia dysfunction.

BACKGROUND: Equine pituitary pars intermedia dysfunction (PPID) is treated with daily pergolide therapy. Owner compliance and its effect on PPID control have not been previously investigated. METHODS: Clinical records were searched to identify the sample of animals with PPID treated with pergolide from 2016 to 2019. The signalment was noted and the dose of pergolide received calculated. Animals were classified as compliant (receiving ≥90% of the veterinarian recommended dose of pergolide) or non-compliant, and as controlled (follow-up basal adrenocorticotrophic hormone concentrations within the reference range) or not. RESULTS: In total, 110 animals were included. The majority (85%) were ≥16 years (mean ± SD 19.8 ± 4.4 years); the most common breeds were Cob (18%), Thoroughbred (16%) and Welsh (15%); 37% were female and 63% male. Overall, 48% were compliant and 52% non-compliant. There was no significant effect of compliance on laboratory control. Of those that were compliant, 74% were controlled, while 67% of non-compliant animals were controlled. Univariable analysis revealed a significant (p < 0.001) effect of age and breed on compliance and control, and of sex on control. On multivariable analysis, only age (compliance) and breed (compliance and control) were retained in the final model. CONCLUSION: Only half of animals received the recommended pergolide dose; however, this did not affect laboratory control of PPID.

Comparison of immunofluorescence and chemiluminescence assays for measuring ACTH in equine plasma.

BACKGROUND: The chemiluminescence (CL) and immunofluorescence (IF) assays yield different results for basal adrenocorticotropin hormone concentrations [ACTH] in pony plasma. It is unclear whether this difference also occurs in basal samples from horses or samples from ponies following thyrotropin-releasing hormone (TRH) stimulation. OBJECTIVES: To compare the results of [ACTH] analysis by CL and IF methods in basal samples from horses and pony samples following TRH stimulation. STUDY DESIGN: Method comparison. METHODS: Plasma [ACTH] was measured concurrently using CL and IF methods in 12 ponies (basal and post-TRH stimulation) in November and basal samples from horses (n = 45; November and May). RESULTS: CL and IF methods yielded different results (P < .01). The median difference (CL-IF) (95% CI) for ponies was 5.9 (0.1-7.5) pg/mL at baseline and 227.9 (61-1001) pg/mL post TRH; and horses 1.9 (1.1-5.4) pg/mL in November and 9.4 (8.2-11.5) pg/mL in May, at baseline. Correlation was good in ponies at baseline (R = 0.80, P = .003) but not post-TRH, and good in horses in November and May (R = 0.68 and 0.71, P < .001). Bland-Altman analysis demonstrated moderate bias and wide 95% limits of agreement (95% LOA) in ponies at baseline (bias 5.5 pg/mL; 95% LOA -9.9 to 20.9 pg/mL) and horses in May (bias 10.6 pg/mL; 95% LOA -9 to 30.3 pg/mL) and very large bias and wide 95% LOA in ponies post-TRH (bias 477 pg/mL; 95% LOA -633 to 1587 pg/mL). Using CL cut-offs of >29 and >110 pg/mL, agreement was moderate (ƙ = 0.67) and very good (ƙ = 0.82) for binary classification of PPID in ponies at baseline and post-TRH; and good (ƙ = 0.73) for horses in November, but poor (ƙ = 0.40) in May. MAIN LIMITATIONS: Limited numbers of horses with [ACTH] above threshold values. CONCLUSIONS: The assays yielded different absolute values, particularly in post-TRH samples from ponies, suggesting TRH stimulates secretion of cross-reacting peptides other than ACTH. Agreement for binary classification for PPID was moderate to good, except in basal samples from horses in May.

Correlation between l-lactate and glucose concentrations and body condition score in healthy horses and ponies.

BACKGROUND: Blood l-lactate and glucose concentrations were higher in ponies with gastrointestinal disease than in horses, possibly because of differences in body condition (BC). OBJECTIVES: To investigate whether l-lactate and glucose concentrations correlate with BC and differ between healthy horses and ponies. ANIMALS: Systemically healthy client-owned ponies (n = 101) and horses (n = 51). METHODS: Prospective observational study. Breed, weight, height, and subjective and objective measures of BC were recorded and l-lactate and glucose concentrations were measured. Correlations between l-lactate and glucose concentrations and BC were established. The association between l-lactate concentrations, equid type (pony or horse), BC, age, and glucose concentrations was investigated using a multivariable model. RESULTS: Weak but significant (P = .001) negative correlations were detected between l-lactate concentration and average BC score (r = -0.29), heart girth:height ratio (r = -0.27), and age (r = -0.27). Glucose concentrations were significantly (P < .001) positively correlated with neck length:heart girth ratio (r = 0.37) and heart girth:height ratio (r = 0.31). l-lactate and glucose concentrations were weakly correlated (r = 0.15; P = .04). In the final multivariable model, age (-0.02 ± 0.006; P = .001) and heart girth:height ratio (-1.74 ± 0.53; P = .001) were significantly associated with the natural logarithm of l-lactate concentration (Lnl-Lactate). This represents a 2% decrease in l-lactate concentration per year increase in age and 10% decrease in l-lactate concentration per 0.06 unit increase in heart girth:height ratio. CONCLUSIONS AND CLINICAL IMPORTANCE: In healthy horses and ponies, age and BC significantly influence l-lactate concentrations.

ECEIM consensus statement on equine metabolic syndrome.

Equine metabolic syndrome (EMS) is a widely recognized collection of risk factors for endocrinopathic laminitis. The most important of these risk factors is insulin dysregulation (ID). Clinicians and horse owners must recognize the presence of these risk factors so that they can be targeted and controlled to reduce the risk of laminitis attacks. Diagnosis of EMS is based partly on the horse's history and clinical examination findings, and partly on laboratory testing. Several choices of test exist which examine different facets of ID and other related metabolic disturbances. EMS is controlled mainly by dietary strategies and exercise programs that aim to improve insulin regulation and decrease obesity where present. In some cases, pharmacologic aids might be useful. Management of an EMS case is a long-term strategy requiring diligence and discipline by the horse's carer and support and guidance from their veterinarians.

The effect of tumour necrosis factor-α and insulin on equine digital blood vessel function in vitro.

OBJECTIVE AND DESIGN: Insulin and inflammatory cytokines may be involved in equine laminitis, which might be associated with digital vascular dysfunction. This study determined the effects of TNF-α and insulin on the endothelial-dependent relaxant responses of equine digital blood vessels and on equine digital vein endothelial cell (EDVEC) cGMP production. MATERIAL: Isolated rings of equine digital arteries (EDAs) and veins (EDVs) were obtained and EDVECs were cultured from horses euthanized at an abattoir. METHODS: The effect of incubation with TNF-α (10 ng/ml) and/or insulin (1,000 µIU/ml) for 1.5 h or overnight under hyperoxic and hypoxic conditions on carbachol (endothelium-dependent) induced relaxation was assessed. The time course and concentration dependency of the effect of TNF-α and the effect of insulin (1,000 µIU/ml) on EDVEC cGMP production was determined. RESULTS: Incubation of EDAs overnight with TNF-α under hypoxic conditions resulted in endothelial-dependent vascular dysfunction. EDVs produced a more variable response. TNF-α increased EDVEC cGMP formation in a time- and concentration-dependent manner. Insulin had no significant effects. CONCLUSIONS: There is a mismatch between the results obtained from isolated vessel rings and cultured endothelial cells suggesting TNF-α may reduce the biological effect of NO by reducing its bioavailability rather than its formation, leading to endothelial cell dysregulation.

Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum.

OBJECTIVE: To evaluate a human radioimmunoassay (RIA) and equine and high-range porcine (hrp) species-specific ELISAs for the measurement of high serum insulin concentrations in ponies. SAMPLES: Serum samples from 12 healthy nonobese ponies (7 clinically normal and 5 laminitis prone; 13 to 26 years of age; 11 mares and 1 gelding) before and after glucose, insulin, and dexamethasone administration. PROCEDURES: Intra-and interassay repeatability, freeze-thaw stability, dilutional parallelism, and assay agreement were assessed. RESULTS: Assay detection limits were as follows: RIA, < 389 µU/mL; equine ELISA, < 175 µU/mL; and hrp ELISA, 293 to 8,775 µU/mL. Mean ± SD intra- and interassay repeatability were respectively as follows: RIA, 6.5 ± 5.1 % and 74 ± 3.4%; equine ELISA, 10.6 ± 11.0% and 9.0 ± 4.6%; and hrp ELISA, 19.9 ± 172% and 173 ± 16.6%. Freezing and thawing affected measured concentrations. Dilutional parallelism in the RIA was only evident when insulin-depleted equine serum was used as a diluent (percentage recovery, 95.7 ± 274%); in the ELISAs, dilutional parallelism was observed when a zero calibrator was used. Agreement between RIA and equine ELISA results was good for samples containing concentrations < 175 µU of insulin/mL (bias, -18.5 ± 25.5 µU/mL; higher in RIA). At higher concentrations, assay agreement was poor between RIA and equine ELISA results (bias, -185.3 ± 98.7 µU/mL) and between RIA and hrp ELISA results (bias, 25.3 ± 183.0 µU/mL). CONCLUSIONS AND CLINICAL RELEVANCE: Agreement among results of the 3 assays was variable, and dilutional parallelism was only evident with the RIA when insulin-depleted equine serum was tested. Caution is recommended when evaluating high insulin concentrations measured with the RIA or ELISAs.