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.

Development of a body condition index to estimate adiposity in ponies and horses from morphometric measurements.

BACKGROUND: There is a high prevalence of obesity in ponies and pleasure horses. This may be associated with equine metabolic syndrome and an increased risk of laminitis. Body condition scoring (BCS) systems are widely used but are subjective and not very sensitive. OBJECTIVES: To derive a body condition index (BCI), based on objective morphometric measurements, that correlates with % body fat. STUDY DESIGN: Retrospective cohort study. METHODS: Morphometric measurements were obtained from 21 ponies and horses in obese and moderate body condition. Percentage body fat was determined using the deuterium dilution method and the BCI was derived to give the optimal correlation with body fat, applying appropriate weightings. The index was then validated by assessing inter-observer variation and correlation with % body fat in a separate population of Welsh ponies; and finally, the correlation between BCI and BCS was evaluated in larger populations from studies undertaken in Australia, the United Kingdom and the United States. RESULTS: The BCI correlated well with adiposity in the ponies and horses, giving a Pearson r value of 0.74 (P < 0.001); however, it was found to slightly overestimate the % body fat in leaner animals and underestimate in more obese animals. In field studies, the correlation between BCI and BCS varied particularly in Shetlands and miniature ponies, presumably due to differences in body shape. MAIN LIMITATIONS: Further work may be required to adapt the BCI to a method that is more applicable for Shetlands and miniature ponies. CONCLUSIONS: This BCI was able to provide an index of adiposity which compared favourably with condition scoring in terms of accuracy of estimating adiposity; and was more consistent and repeatable when used by inexperienced assessors. Therefore, this may be a useful tool for assessing adiposity; and may be more sensitive than condition scoring for tracking weight gain or weight loss in individual animals.

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.

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.

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.

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.