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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 (r s [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., Competing Interests: Declaration of Competing Interest None of the authors has a financial or personal relationship with other people or organisations that could inappropriately influence or via the content of the paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

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., (© 2023 Mars Horsecare UK Ltd and The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Several tests have been advocated for diagnosis of insulin dysregulation (ID). Tests using simple sugars may not reflect the response to naturally ingested carbohydrates. This study aimed to evaluate agreement between the oral glucose test (OGT), the oral sugar test (OST), a novel oral test using a proprietary cereal (WEET), the IV combined glucose-insulin tolerance test (CGIT) and fasted basal insulin (FI) for diagnosis of ID. Each of the five tests above was performed on a group of six normal and six insulin dysregulated mixed-breed ponies in a randomised crossover study. Area under the curve (AUC) and maximum concentration of insulin from OGT, OST and WEET showed strong to very strong bivariate correlations (r = 0.85-0.94, and r = 0.87-0.92, respectively; P ≤ 0.001) and were significantly different between tests (associated with dose of carbohydrate) and between CGIT-positive and -negative ponies. Dichotomous results showed substantial agreement between OST and both WEET (κ = 0.65; P = 0.02) and OGT (κ = 0.67; P = 0.01) and between CGIT and both OST (κ = 0.63; P = 0.03) and OGT (κ = 0.67; P = 0.01), and no agreement between FI, which had low sensitivity, and all other tests (κ = 0.15 - 0.31; P > 0. 05). Palatability of WEET was variable, resulting in one pony being excluded for analysis of WEET data. Further work on development of an oral test using a more palatable feedstuff and appropriate cut-offs or diagnostic thresholds for tests of ID is warranted., Competing Interests: Declaration of Competing Interest None of the authors has a financial or personal relationship with other people or organisations that could inappropriately influence or via the content of the paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Native ponies are at increased risk of obesity and metabolic perturbations, such as insulin dysregulation (ID), a key risk factor for endocrinopathic laminitis. Management and feeding practices can be adapted to maintain healthy body condition and support metabolic health, but owners may inadvertently provide their ponies with inappropriate management leading to obesity and exacerbating risk of metabolic disease. Adoption of preventative weight management approaches (WMAs), including regular monitoring of body condition, providing appropriate preserved forage, promoting seasonal weight loss, and using exercise accordingly, are key in supporting native ponies' metabolic health. The factors influencing the adoption of WMAs, such as owners' experience and confidence, require exploration. The aim of the current study was to understand factors influencing owners' likelihood to undertake certain WMAs, to develop our understanding of suitable intervention targets. A total of 571 responses to an online cross-sectional questionnaire were analysed. Mediation analysis revealed that whilst long term (≥20 years) experience caring for native ponies was associated with owners increased, self-reported confidence in identifying disease and managing their native ponies, this did not translate to an increased likelihood of implementing WMAs. Conversely, respondents who managed ponies with dietary requirements related to obesity, laminitis, or equine metabolic syndrome were more likely to use WMAs related to feeding, seasonal weight management and exercise. Owner confidence was assessed and rejected as a mediator of the relationship between experience and WMA use. These results highlight the need for further work that elucidates the pathways leading owners to undertake action against obesity without the need for ponies to develop overt disease, as well as suggesting a need for long term managers of native ponies to update management practices with preventative care as the focus., Competing Interests: Co-author PH is employed by the funding organization. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Ward et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The equine faecal microbiota is often assessed as a proxy of the microbial community in the distal colon, where the microbiome has been linked to states of health and disease in the horse. However, the microbial community structure may change over time if samples are not adequately preserved. This study stored equine faecal samples from n = 10 horses in four preservation treatments at room temperature for up to 150 h and assessed the resulting impact on microbial diversity and the differential abundance of taxa. Treatments included "COLD" (samples packaged with a cool pack), "CLX" (2% chlorhexidine digluconate solution), "NAP" (nucleic acid preservation buffer), and "FTA" (Whatman FTA™ cards). The samples were assessed using 16S rRNA gene sequencing after storage for 0, 24, 72, and 150 h at room temperature under the different treatments. The results showed effective preservation of diversity and community structure with NAP buffer but lower diversity ( p = 0.001) and the under-representation of Fibrobacterota in the FTA card samples. The NAP treatment inhibited the overgrowth of bloom taxa that occurred by 72 h at room temperature. The COLD, CLX, and NAP treatments were effective in preserving the faecal microbiota for up to 24 h at room temperature, and the CLX and NAP treatments improved the yield of Patescibacteria and Fibrobacterota in some cases. The cold and CLX treatments were ineffective in preventing community shifts that occurred by 72 h at room temperature. These findings demonstrate the suitability of the COLD, NAP, and CLX treatments for the room temperature storage of equine faeces for up to 24 h and of NAP buffer for up to 150 h prior to processing., Competing Interests: The authors declare no conflicts of interest. With the exception of PH (employed by the funding organization), the funding organization did not have any additional role in the conceptualization, methodology, investigation, data curation, formal analysis, decision to publish, or preparation of the manuscript. PH was involved in study design, data interpretation, and manuscript preparation.

Caroline McGregor Argo argues that the new vet school at SRUC will not just be 'more of the same', and that instead it will have a clear remit to produce vets with attributes aligned with society's priority needs., (© 2021 British Veterinary Association.)

The COVID-19 pandemic continues to impact human health and welfare on a global level. In March 2020, stringent national restrictions were enforced in the UK to protect public health and slow the spread of the SARS-CoV-2 virus. Restrictions were likely to have resulted in collateral consequences for the health and welfare of horses and ponies, especially those at risk of obesity and laminitis and this issue warranted more detailed exploration. The current study utilised qualitative methodology to investigate the implications of COVID-19 related policies upon equine management and welfare with a focus on horses and ponies at risk of laminitis and obesity. Twenty-four interviews with five sub-groups of key equestrian welfare stakeholders in the UK were conducted between May and July of 2020 to understand the challenges facing equine welfare in the context of laminitis and obesity susceptible animals. Thematic analysis revealed lockdown-associated factors with the potential to compromise welfare of horses and ponies at risk of obesity and laminitis. These included: disparate information and guidance, difficulties enacting public health measures in yard environments, and horses having reduced exercise during the pandemic. Positive examples of clear and consistent information sharing by farriers were reported to have improved horse owner understanding of routine hoof care during lockdown. Analysis suggested that the recommendations for supporting the management-based needs of horses under reduced supervision were not clearly defined, or were not sufficiently disseminated, across the equine industry. These findings support the development of guidelines specific to the care of horses and ponies at risk of obesity and laminitis through collaborative input from veterinary and welfare experts, to reduce the negative impacts of future lockdown events in the UK., Competing Interests: Co-author PH is employed by the funding organization. This does not alter our adherence to PLOS ONE policies on sharing data and materials. All other authors declare that they have no competing interests.

Bacteria residing in the gastrointestinal tract of mammals are crucial for the digestion of dietary nutrients. Bacterial community composition is modified by age and diet in other species. Although horses are adapted to consuming fibre-based diets, high-energy, often high-starch containing feeds are increasingly used. The current study assessed the impact of age on the faecal bacteriome of ponies transitioning from a hay-based diet to a high-starch diet. Over two years, 23 Welsh Section A pony mares were evaluated (Controls, 5-15 years, n = 6/year, 12 in total; Aged, ≥19 years, n = 6 Year 1; n = 5 Year 2, 11 in total). Across the same 30-week (May to November) period in each year, animals were randomly assigned to a 5-week period of study and were individually fed the same hay to maintenance (2% body mass as daily dry matter intake) for 4-weeks. During the final week, 2g starch per kg body mass (micronized steam-flaked barley) was incorporated into the diet (3-day transition and 5 days at maximum). Faecal samples were collected for 11 days (final 3 days hay and 8 days hay + barley feeding). Bacterial communities were determined using Ion Torrent Sequencing of amplified V1-V2 hypervariable regions of 16S rRNA. Age had a minimal effect on the bacteriome response to diet. The dietary transition increased Candidatus Saccharibacteria and Firmicutes phyla abundance and reduced Fibrobactres abundance. At the genera level, Streptococcus abundance was increased but not consistently across individual animals. Bacterial diversity was reduced during dietary transition in Streptococcus 'responders'. Faecal pH and VFA concentrations were modified by diet but considerable inter-individual variation was present. The current study describes compositional changes in the faecal bacteriome associated with the transition from a fibre-based to a high-starch diet in ponies and emphasises the individual nature of dietary responses, which may reflect functional differences in the bacterial populations present in the hindgut., Competing Interests: Co-authors PH and CB are employed by the funding organization, WALTHAM (https://www.waltham.com/). Co-author AD is employed by ChesterGates Veterinary Specialists CVS Ltd. This does not alter our adherence to PLOS ONE policies on sharing data and materials. All other authors declare that they have no competing interests.

Background: Obesity is an important equine welfare issue. Whilst dietary restriction is the most effective weight-loss tool, individual animals range in their weight-loss propensity. Gastrointestinal-derived bacteria play a fundamental role in host-health and have been associated with obesity and weight-loss in other species. This study evaluated the faecal microbiome (next-generation sequencing of 16S rRNA genes) of 15 obese Welsh Mountain pony mares, in the same 11-week period across 2 years (n = 8 Year 1; n = 7 Year 2). Following a 4-week acclimation period (pre-diet phase) during which time individuals were fed the same hay to maintenance (2% body mass (BM) as daily dry matter (DM) intake), animals underwent a 7-week period of dietary restriction (1% BM hay as daily DM intake). Faeces were sampled on the final 3 days of the pre-diet phase and the final 3 days of the dietary restriction phase. Bacterial communities were determined using Next Generation Sequencing of amplified V1-V2 hypervariable regions of bacterial 16S rRNA., Results: Losses in body mass ranged from 7.11 to 11.59%. Changes in the faecal microbiome composition following weight-loss included a reduction in the relative abundance of Firmicutes and Tenericutes and a reduction in indices of bacterial diversity. Pre-diet diversity was negatively associated with weight-loss. Pre-diet faecal acetate concentration was a strong predictor of subsequent weight-loss and negatively associated with Sphaerochaeta (Spirochaetes phylum) abundance. When animals were divided into 3 groups (high, mid, low) based overall weight loss, pre-diet bacterial community structure was found to have the greatest divergence between the high and low weight-loss groups (R = 0.67, p <  0.01), following PERMANOVA and ANOSIM analysis., Conclusions: Weight-loss in this group of ponies was associated with lower pre-diet faecal bacterial diversity and greater pre-diet acetate concentration. Overall, these data support a role for the faecal microbiome in weight-loss propensity in ponies and provide a baseline for research evaluating elements of the faecal microbiome in predicting weight-loss success in larger cohorts.

Gastrointestinal microbial communities are increasingly being implicated in host susceptibilities to nutritional/metabolic diseases; such conditions are more prevalent in obese and/or older horses. This controlled study evaluated associations between host-phenotype and the fecal microbiome / metabolome. Thirty-five, Welsh Mountain pony mares were studied across 2 years (Controls, n = 6/year, 5-15 years, Body Condition Score (BCS) 4.5-6/9; Obese, n = 6/year, 5-15 years, BCS > 7/9; Aged, n = 6 Year 1; n = 5 Year 2, ≥19 years old). Animals were individually fed the same hay to maintenance (2% body mass as daily dry matter intake) for 2 (aged / obese) or 4 (control), 4-week periods in a randomized study. Outset phenotype was determined (body fat%, markers of insulin sensitivity). Feces were sampled on the final 3 days of hay feeding-periods and communities determined using Next Generation Sequencing of amplified V1-V2 hypervariable regions of bacterial 16S rRNA. Copy numbers for fecal bacteria, protozoa and fungi were similar across groups, whilst bacterial diversity was increased in the obese group. Dominant bacterial phyla in all groups were Bacteroidetes > Firmicutes > Fibrobacter . Significant differences in the bacterial communities of feces were detected between host-phenotype groups. Relative to controls, abundances of Proteobacteria were increased for aged animals and Bacteroidetes , Firmicutes , and Actinobacteria were increased for obese animals. Over 500 bacterial operational taxonomic units (OTUs) differed significantly between host-phenotype groups. No consistent pattern of changes in discriminant OTUs between groups were maintained across groups and between years. The core bacterial populations contained 21 OTUs, 6.7% of recovered sequences. Distance-based Redundancy Analyses separated fecal bacterial communities with respect to markers of obesity and insulin dysregulation, as opposed to age. Host-phenotype had no impact on the apparent digestibility of dietary GE or DM, fecal volatile fatty acid concentrations or the fecal metabolome (FT-IR). The current study demonstrates that host-phenotype has major effects on equine fecal microbial population structure. Changes were predominantly associated with the obese state, confirming an obesity-associated impact in the absence of nutritional differences. Clear biomarkers of animal-phenotype were not identified within either the fecal microbiome or metabolome, suggesting functional redundancy within the gut microbiome and/or metabolome.

Reduction of the hyperinsulinaemic response to feeding is central to the management of insulin dysregulation (ID). The aim of this study was to compare insulinaemic and glycaemic responses to soaked hay, dry hay and haylage in ponies. Twelve ponies of mixed breeds were maintained under identical management conditions. A randomised four-way crossover trial was conducted, in which fasted animals were fed a meal of 0.25% body weight as dry matter intake soaked hay, dry hay or haylage, or administered an oral glucose test (OGT). Blood glucose and serum insulin concentrations were measured before and at 2h following OGT, and regularly for 5h following forage meals. Median and interquartile range (IQR) area under the curve (AUC) for insulin (AUC i ) was greater for haylage (median 6495; IQR 17352) vs. dry hay (2932; IQR 5937; P=0.019) and soaked hay (1066; IQR 1753; P=0.002), and greater for dry hay vs. soaked hay (P=0.002). The AUC for glucose (AUC g ) was lower for soaked hay (1021; IQR 99) vs. dry hay (1075; IQR 105; P=0.002) and haylage (1107; IQR 221; P=0.003). Six ponies were classified as having ID based on the OGT. AUC i was greater in ID vs. non-ID ponies after all forages. In contrast, there was no detectable effect of ID status on AUC g . On an equivalent dry matter basis, soaked hay produced the lowest insulinaemic and glycaemic responses to feeding, while haylage produced the highest responses. The insulinaemic effects of all forages were greater in ponies with ID. These data support the practice of soaking hay with water to reduce postprandial insulinaemic responses in ponies., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Body condition score and percent body fat (BF; %) of horses are positively correlated with reproductive efficiency and are indicative of metabolic issues. However, BF in horses may be poorly predicted because current procedures are either subjective or dependent on one anatomical location. Therefore, the objectives of the current study were to compare 2 methods of predicting BF using rump fat thickness (RFT) and deuterium oxide (DO) dilution with actual tissue fat analysis by near-infrared spectroscopy (NIRS) in stock-type horses and to identify the relationship between BF and BCS. Twenty-four stock-type horses were selected to be humanely euthanized based on 3 primary criteria: geriatric, crippled, and/or unsafe. Approximately 20 h before slaughter, horses were weighed and BCS assessed to be 1 ( = 1; 433 kg), 2 ( = 1; 415 kg), 3 ( = 1; 376 kg), 4 ( = 7; 468 ± 13 kg), 5 ( = 10; 455 ± 11 kg), and 6 ( = 4; 493 ± 12 kg) and RFT was measured using ultrasonography. Blood samples were collected immediately before and 4 h after DO infusion (0.12 g/kg BW). Deuterium oxide concentration of plasma was determined by gas isotope ratio mass spectrometry. Horses were housed in a dry lot overnight before being individually sedated (1.1 mg xylazine/kg BW) and anesthetized using a jugular venipuncture (2.2 mg ketamine/kg BW), and potassium chloride (KCl) solution was administered to cease cardiac function before exsanguination. After euthanasia, horse carcasses were processed and dissected and tissues were collected for NIRS analysis. Body fat predicted by DO dilution was correlated with BF measured by NIRS analysis on various weight bases ( = 0.76 to 0.81, < 0.001), whereas no correlation was observed for BF predicted by RFT. A paired -test indicated that both DO dilution and RFT overestimated BF by 2.48 to 3.26% ( < 0.001) and 5.81 to 6.59% ( < 0.001), respectively, compared with NIRS analysis. Body condition scores affected BF measured by NIRS analysis ( ≤ 0.038) and predicted by DO dilution ( < 0.001) and RFT ( = 0.042). The current study provided evidence that DO dilution was a better prediction method for BF than RFT and that BCS may serve as a BF predictor in stock-type horses.

Twelve mature (aged 5-16 years) horses and ponies of mixed breed and type were fed restricted (1.25% BM Dry matter) quantities of one of two fiber based diets formulated to be iso-caloric. Diet 1 comprised of 0.8% body mass (BM) of chaff based complete feed plus 0.45% BM low energy grass hay (the same hay used for both diets). Diet 2 comprised 0.1% BM of a nutrient balancer plus 1.15% BM grass hay. Fecal samples were collected at week 10 and week 16. DNA was extracted and the V1-V2 regions of 16SrDNA were 454-pyrosequenced to investigate the bacterial microbiome of the horse. The two most abundant phyla found in both diets and sampling periods were the Firmicutes and Bacteroidetes. There was a clear reduction in Bacteroidetes with a concordant increase in Firmicutes over time. There was a limited degree of stability within the bacterial community of the hindgut of horses, with 65% of bacteria retained, over a 6 week period whilst on a uniform diet. The presence of a core community defined by being present in all samples (each animal/diet combination) included in the study and being present at 0.1% relative abundance (or greater) was identified. In total 65 operational taxonomic units (OTUs) were identified that fit the definition of core making up 21-28% of the total sequences recovered. As with total population the most abundant phyla were the Bacteroidetes followed by the Firmicutes , however there was no obvious shift in phyla due to period. Indeed, when the relative abundance of OTUs was examined across diets and periods there was no significant effect of diet or period alone or in combination on the relative abundance of the core OTUs.

Anatomically distinct adipose tissues represent variable risks to metabolic health in man and some other mammals. Quantitative-imaging of internal adipose depots is problematic in large animals and associations between regional adiposity and health are poorly understood. This study aimed to develop and test a semi-quantitative system (EQUIFAT) which could be applied to regional adipose tissues. Anatomically-defined, photographic images of adipose depots (omental, mesenteric, epicardial, rump) were collected from 38 animals immediately post-mortem. Images were ranked and depot-specific descriptors were developed (1 = no fat visible; 5 = excessive fat present). Nuchal-crest and ventro-abdominal-retroperitoneal adipose depot depths (cm) were transformed to categorical 5 point scores. The repeatability and reliability of EQUIFAT was independently tested by 24 observers. When half scores were permitted, inter-observer agreement was substantial (average κw: mesenteric, 0.79; omental, 0.79; rump 0.61) or moderate (average κw; epicardial, 0.60). Intra-observer repeatability was tested by 8 observers on 2 occasions. Kappa analysis indicated perfect (omental and mesenteric) and substantial agreement (epicardial and rump) between attempts. A further 207 animals were evaluated ante-mortem (age, height, breed-type, gender, body condition score [BCS]) and again immediately post-mortem (EQUIFAT scores, carcass weight). Multivariable, random effect linear regression models were fitted (breed as random effect; BCS as outcome variable). Only height, carcass weight, omental and retroperitoneal EQUIFAT scores remained as explanatory variables in the final model. The EQUIFAT scores developed here demonstrate clear functional differences between regional adipose depots and future studies could be directed towards describing associations between adiposity and disease risk in surgical and post-mortem situations.

Contentious issues in calf rearing include milk feeding level and single versus group housing. The current study was performed on a high-producing 170 Holstein cow dairy farm to investigate the impact of nutrition and housing on disease incidence. Calves (n=100) were allocated in birth order to one of two commonly used feeding strategies. Group A calves were group housed from birth and fed ad libitum milk replacer (MR) via a computerised machine using a single teat, with weaning commencing at 63 days. Group R calves were initially housed in individual pens receiving 2.5 litres of MR twice daily via a bucket until three weeks of age when they were group housed and fed 3 litres of MR twice daily via a group trough with weaning commencing at 56 days. In total, 80 (80 per cent) calves suffered from at least one incident of disease during the period from birth to 12 weeks. Group A calves had a greater risk of disease than group R calves (diarrhoea: OR 3.86 (95 per cent CI 1.67 to 8.9); pneumonia: OR 5.80 (95 per cent CI 2.33 to 14.44)). There was a 5.1 per cent incidence of failure of passive transfer of Ig assessed via measurement of plasma total protein concentrations at 48 hours of age. It is hypothesised that the increased diarrhoea risk in group A calves was most likely associated with group housing, while the increased pneumonia risk was associated with the use of a single teat allowing increased transmission of pathogens from calf to calf., (British Veterinary Association.)

Leisure animals now comprise the majority of working horses in industrialized nations; a shift that has decreased workloads yet improved veterinary care and lifetime health. Although many horses now progress well into their 20s without any requirement for dietary modification, age-related changes are insidious, and older animals benefit from regular veterinary monitoring to identify, address, and ameliorate the inevitable onset of age-related "disease." Basal metabolic rate decreases with age; older animals expend less energy on controlled exercise, and there can be an increased propensity toward the development of obesity, which needs to be recognized and managed., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The addition of hay soaking to current nutritional advice for weight loss management for equine obesity lacks clinical evidence. Twelve overweight/obese horses and ponies were used to test the hypothesis that feeding soaked hay at 1.25% of body mass (BM) daily as dry matter (DM) before soaking would elicit weight losses within the target 0.5-1.0% of BM weekly. Six animals were used to evaluate the impact of nutrient-leaching on the digestibility and daily intakes of dietary energy and nutrients. Soaked hay DM was corrected in accordance with the 'insoluble' ADF content of fresh and soaked hays. The ADF-based method was validated using a test-soaking protocol. Animals fed soaked hay for 6 weeks lost 0.98 ± 0.10% of BM weekly. The most weight loss sensitive animal lost ~2% of BM weekly. Soaking hay did not alter DM gross energy concentrations, incurred losses of water soluble carbohydrates (WSC) and ash and increased acid detergent fibre (ADF) concentrations. Digestibilities of GE, DM, ash and WSC were unaltered but soaking increased uncorrected values for crude protein (+12%) and ADF (+13.5%) digestibility. Corrected DM provision was only 1% of BM daily, providing 64% of maintenance DE requirements, a 23.5% increase in the intended magnitude of energy restriction. Hay soaking leached nutrients, reduced DM and DE provision and was associated with accelerated weight losses over those expected had fresh-hay been fed to the same level. The ADF-based method will allow the predictive evaluation of individual hays to direct feeding management and prevent inadvertently severe DM and energy restriction., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Obesity is a widespread problem across the leisure population of horses and ponies in industrialised nations. Skeletal muscle is a major contributor to whole body resting energy requirements and communicates with other tissues through the secretion of myokines into the circulation. Myostatin, a myokine and negative regulator of skeletal muscle mass, has been implicated in obesity development in other species. This study evaluated gene and protein expression of myostatin and its receptor, ActRIIB in adipose tissues and skeletal muscles and serum myostatin concentrations in six lean and six obese animals to explore putative associations between these factors and obesity in horses and ponies. Myostatin mRNA expression was increased while ActRIIB mRNA was decreased in skeletal muscles of obese animals but these differences were absent at the protein level. Myostatin mRNA was increased in crest fat of obese animals but neither myostatin nor ActRIIB proteins were detected in this tissue. Mean circulating myostatin concentrations were significantly higher in obese than in lean groups; 4.98 ng/ml (±2.71) and 9.00 ng/ml (±2.04) for the lean and obese groups, respectively. In addition, there was a significant positive association between these levels and myostatin gene expression in skeletal muscles (average R2 = 0.58; p<0.05). Together, these results provide further basis for the speculation that myostatin and its receptor may play a role in obesity in horses and ponies.

Obesity, a major concern for equine welfare, is highly prevalent in the leisure horse population. Skeletal-muscle and adipose tissues are important determinants of maintenance energy requirements. The myostatin and perilipin pathways play key roles in the regulation of muscle mass and lipolysis respectively and have both been associated with obesity predisposition in other mammalian species. High quality samples, suitable for molecular biology, are an essential prerequisite for detailed investigations of gene and protein expression. Hence, this study has evaluated a) the post-mortem stability of RNA extracted from skeletal-muscle and adipose-tissues collected under commercial conditions and b) the tissue-specific presence of myostatin, the moystatin receptor (activin receptor IIB, ActRIIB), follistatin and perilipin, genes and proteins across a range of equine tissues. Objectives were addressed using tissues from 7 Thoroughbred horses presented for slaughter at a commercial abattoir; a) samples were collected at 7 time-points from Masseter muscle and perirenal adipose from 5 minutes to 6 hours post-mortem. Extracted RN was appraised by Optical Density analysis and agarose-gel electrophoresis. b) Quantitative real time PCR and Western Blotting were used to evaluate gene and protein expression in anatomically-defined samples collected from 17 tissues (6 organs, 4 skeletal muscles and 7 discrete adipose depots). The results indicate that, under the present collection conditions, intact, good quality RNA could be extracted from skeletal-muscle for up to 2 hours post-mortem. However, RNA from adipose tissue may be more susceptible to degradation/contamination and samples should be collected no later than 30 minutes post-mortem. The data also show that myostatin and ActRIIB genes and proteins were almost exclusively expressed in skeletal muscle. The follistatin gene showed a more diverse gene expression profile, with expression evident in several organs, adipose tissue depots and skeletal muscles. Perilipin gene and protein were almost exclusively expressed by adipose tissue.

The horse, as a hindgut fermenter, is reliant on its intestinal bacterial population for efficient diet utilisation. However, sudden disturbance of this population can result in severe colic or laminitis, both of which may require euthanasia. This study therefore aimed to determine the temporal stability of the bacterial population of faecal samples from six ponies maintained on a formulated high fibre diet. Bacterial 16S rRNA terminal restriction fragment length polymorphism (TRFLP) analyses of 10 faecal samples collected from 6 ponies at regular intervals over 72 hour trial periods identified a significant pony-specific profile (P<0.001) with strong stability. Within each pony, a significantly different population was found after 11 weeks on the same diet (P<0.001) and with greater intra-individual similarity. Total short chain fatty acid (SCFA) concentration increased in all ponies, but other changes (such as bacterial population diversity measures, individual major SCFA concentration) were significant and dependent on the individual. This study is the first to report the extent of stability of microbes resident in the intestinal tract as represented with such depth and frequency of faecal sampling. In doing so, this provides a baseline from which future trials can be planned and the extent to which results may be interpreted.

Few studies have examined the effect of dietary restriction in horses with equine metabolic syndrome (EMS). This study aimed to determine improvements in insulin sensitivity following dietary restriction for 6 weeks, and to determine if the improvement would be greater in horses receiving short-chain fructo-oligosaccharides (sc-FOS). Dietary management involved feeding grass hay, restricted to 1.25% of body mass (BM) as daily dry matter intake and soaked in cold water prior to feeding, with the addition of a vitamin and mineral nutraceutical supplement with or without the addition of sc-FOS (10 g/100 kg). Soaking the hay resulted in a significant reduction in non-structural carbohydrates (38%, P = 0.01), digestible energy (6.78%, P = 0.01) and water soluble minerals. Following 6 weeks of dietary restriction with soaked grass hay and nutraceutical supplement, horses lost an average of 6.8% BM and showed reductions in body condition score (BCS) and belly circumference. Sensitivity to insulin improved overall, as determined by the total insulin response during the combined glucose insulin test. The magnitude of improvement in insulin sensitivity was associated with the degree of insulin resistance recorded at outset, and the extent of overall losses in BM and BCS, but was independent of the addition of sc-FOS. The nutraceutical supplement was highly palatable and no adverse effects were noted. From the findings of this study a strict dietary program in combination with a specifically designed vitamin and mineral nutraceutical supplement can be recommended to obtain rapid improvements in BM, BCS and insulin sensitivity of animals presenting with EMS., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Evidence-based, weight loss management advice is required to address equine obesity. Changes in body mass (BM), body condition score (BCS), heart (HG) and belly circumference (BG), direct (ultrasonographic) and indirect (D(2)O dilution, bioelectrical impedance analysis [BIA]) measures of body fat as well as indices of insulin resistance (IR) were monitored in 12 overweight (BCS ≥ 7/9) horses and ponies of mixed breed and gender for 16 weeks. Animals were randomly assigned to two groups (Group 1, n=6, BCS 7.6/9 ± 0.6, 489 ± 184.6 kg; Group 2, n=6, BCS 8.1/9 ± 0.6, 479 ± 191.5 kg). Daily dry matter intake (DMI) was restricted to 1.25% BM as one of two, near-isocaloric (DE ∼0.115 MJ/kg BM/day), forage-based diets (Group 1, 0.8% BM chaff-based feed: 0.45% BM hay; Group 2, 1.15% BM hay: 0.1% BM nutrient-balancer). Statistical modelling revealed considerable between-animal heterogeneity in proportional weight losses (0.16-0.55% of Week 1 BM weekly). The magnitude of weight loss resistance (WLR) or sensitivity to dietary restriction was independent of diet or any measured outset variable and was largely (65%) attributed to animal identity. Predicted rates of weight loss decreased over time. BCS and BIA were poor estimates of D(2)O-derived body fat%. Reciprocal changes in depths of retroperitoneal and subcutaneous adipose tissues were evident. Changes in BG were associated with losses in retroperitoneal fat and BM (r(2), 0.67 and 0.79). Indices of IR improved for 9/12 animals by Week 16. For obese animals, weight loss should be initiated by restricting forage DMI to 1.25% BM. Subsequent restriction to 1% BM may be warranted for WLR animals., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Body condition scoring systems were originally developed to quantify flesh cover in food animals and are commonly used to evaluate body fat in Equidae. The relationship between concurrent estimates of body fat content (eTBF%, deuterium oxide dilution; range, 2.7-35.6%) and subjective appraisals of body 'fatness' (body condition score, BCS; range, 1.25-9/9), was investigated in 77 mature horses and ponies. Univariate (UVM, r(2)=0.79) and multivariable (MVM, r(2)=0.86) linear regression models described the association, where BCS and eTBF% were explanatory and outcome variables, respectively. Other measures (age, sex, breed, body mass, ultrasound-generated subcutaneous and abdominal retroperitoneal fat depths, withers height, heart and belly circumferences) were considered as potential confounders but only height, belly circumference and retroperitoneal fat depth remained in the final MVM. The association between BCS and eTBF% was logarithmic. Appraisal of the transformed regression (UVM), actual eTBF% values and 95%CIs of the model forecast, suggested that the power of log-transformed BCS as a predictor of eTBF% decreased as BCS increased. The receiver operating characteristic curve for the prediction of horses with an eTBF% of >20%, suggested that the UVM correctly classified 76% of horses using a 'cut-off' of BCS 6.83/9 (sensitivity, 82.5%; specificity, 70.8%). Negative values for eTBF% were obtained for two thin ponies which were excluded from analyses, and caution is advised in the application of deuterium dilution methodologies where perturbed tissue hydration could be predicted. The data suggest that BCS descriptors may warrant further consideration/refinement to establish more clinically-useful, sub-classifications for overweight/obese animals., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Changes in appetite, body mass (BM), body condition score (BCS), direct (ultrasonographic) and indirect (deuterium oxide dilution technique) measures of body fat were monitored in Welsh Mountain pony mares (n=11, 5-19 years of age) offered ad libitum access to a complete diet (gross energy 16.9±0.07 MJ/kg dry matter) for 12 weeks during summer (n=6; 246±20 kg) and winter (n=5; 219±21 kg). At the outset, each group comprised two thin (BCS 1-3/9), moderate (BCS 4-6/9) and obese (BCS 7-9/9) animals. For ponies that were non-obese at the outset, BM was gained more rapidly (P=0.001) in summer (0.8±0.1 kg/day) than winter (0.6±0.0 kg/day). This was associated with a seasonal increase in dry matter intake (DMI) which became maximal (summer, 4.6±0.3% BM as DMI/day; winter, 3.5±0.1% BM as DMI/day) during the second month. The appetite of the obese ponies was half that reported for non-obese animals in the summer and BM remained constant irrespective of season. Body 'fatness' increased progressively for non-obese but not obese ponies. Body fat content was exponentially associated with increasing BCS but BCSs >6 were not useful indicators of actual body fat. Endogenous circannual mechanisms to suppress winter weight gain were insufficient to prevent the development of obesity in ad libitum fed ponies., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Dietary restriction for the weight-loss management of obese horses limits the natural trickle-feeding behaviour. During feed restriction, wood shavings are often advised as bedding to prevent dietary supplementation from non-feed sources. Data from twelve overweight/obese horses and ponies of mixed breed and sex, bedded on wood shavings during 16 weeks of feed restriction, were retrospectively evaluated. DM intake (DMI) was restricted to 1.25 % of body mass (BM) daily. Animals were randomly assigned to one of two diets (hay/chaff, n 6; hay/balancer meal, n 6). BM was recorded weekly. Feeding behaviour was recorded by continual observation over 24 h during week 15. The apparent digestibility (gross energy (GE), acid-detergent fibre (ADF) and DM) of feed was determined for all animals by total faecal collection (72 h, week 16). Rates of weight loss were independent of diet type, DM (R(2) 0.15), GE (R(2) 0.20) and ADF digestibilities (R(2) 0.18). Despite similar DMI, faecal DM ranged between 0.52 and 1.16 % of BM daily and was associated with wide ranges in apparent digestibility (GE--11.34 to 53.08 %; ADF--50.37 to 42.83 % and DM 2.14 to 57.32 %), which were improbably low for some animals. Apparent digestibilities were associated with DM output (GE R2 0.96; ADF R(2) 0.99 and DM R(2) 0.99) and time spent feeding (GE R(2) 0.62; DM R(2) 0.61 and ADF R(2) 0.59), indicating that feed intake was supplemented with wood shavings in at least five of the twelve animals. Quantities of wood shavings ingested (negligible to >3.0 kg/d) were back-calculated from predicted feed digestibilities. All animals remained healthy. Implications of 'feed-bulking/energy dilution' for feed-restricted animals need further consideration.

Reasons for Performing the Study: Excessive accumulations or depletions of body fat have been associated with increased morbidity and mortality in horses and ponies. An objective, minimally-invasive method to accurately quantify body fat in living animals is required to aid nutritional management and define welfare/performance limits., Objectives: To compare deuterium oxide (D(2) O) dilution-derived estimates of total body water (TBW) and body fat with values obtained by 'gold standard' proximate analysis and cadaver dissection., Hypothesis: D(2) O dilution offers a valid method for the determination of TBW and body fat in equids., Methods: Seven mature (mean ± s.e. 13 ± 3 years, 212 ± 14 kg, body condition scores 1.25-7/9), healthy, Welsh Mountain pony mares, destined for euthanasia (for nonresearch purposes) were used. Blood samples were collected before and 4 h after D(2) O (0.11-0.13 g/kg bwt, 99.8 atom percent excess) administration. Plasma was analysed by gas isotope ratio mass spectrometry following filtration and zinc reduction. After euthanasia, white adipose tissue (WAT) mass was recorded before all body tissues were analysed by proximate chemical analyses., Results: D(2) O-derived estimates of TBW and body fat were strongly associated with proximate analysis- and dissection-derived values (all r(2) >0.97, P≤0.0001). Bland-Altman analyses demonstrated good agreements between methods. D(2) O dilution slightly overestimated TBW (0.79%, limits of agreement (LoA) -3.75-2.17%) and underestimated total body lipid (1.78%, LoA -0.59-4.15%) and dissected WAT (0.72%, LoA -2.77-4.21%)., Conclusions and Potential Relevance: This study provides the first validation of the D(2) O dilution method for the minimally-invasive, accurate, repeatable and objective measurement of body water and fat in living equids., (© 2011 EVJ Ltd.)

Reasons for Performing Study: Evaluation of equine body fat content is important for nutritional and clinical purposes. However, our understanding of total body fat and its regional distribution in the body is sparse. Currently, body fat evaluation relies on the subjective assessment of body condition score (BCS), which has never been validated against 'gold standard' chemical analysis or dissection measurements in ponies., Objectives: To define the relationships between subjective (BCS), objective (morphometric) indices of body fat and 'gold standard' measurements of actual body composition., Hypotheses: BCS and morphometry offer valid, noninvasive methods for determination of body fat in equids., Methods: Seven mature (mean ± s.e. 13 ± 3 years, 212 ± 14 kg, BCS 1.25-7/9), Welsh Mountain pony mares, destined for euthanasia (for nonresearch purposes), were used. For all ponies, body mass (BM), BCS and various morphometric measurements were recorded. Following euthanasia, all ponies were systematically dissected. Discrete white adipose tissue (WAT) depots were independently described. Gross, body chemical composition was determined by proximate analyses., Results: Total somatic soft tissues increased linearly (r(2) = 1.00), whereas body WAT content (1-26% live BM) increased exponentially (r(2) = 0.96), with BCS. WAT was equally distributed between internal and external sites in all animals irrespective of BCS. Nuchal fat was a poor predictor of total WAT (r(2) = 0.66). Periorbital WAT did not alter with BCS (r(2) = 0.01). Heart girth:withers height and ultrasonic retroperitoneal fat depth were closely associated with total, chemically-extracted lipid which comprised 1-29% live BM (r(2) = 0.91 and 0.88, respectively)., Conclusions and Potential Relevance: The exponential relationship between BCS and total body WAT/lipid suggests that BCS is unlikely to be a sensitive index of body fat for animals in moderate-obese states. Morphometric measurements (body girths and retroperitonel fat depth) may be useful to augment subjective BCS systems., (© 2011 EVJ Ltd.)

A total of 318 cows were monitored in the pre-breeding postpartum period for the presence of three production stressors: lameness, subclinical mastitis and body condition score (BCS) loss. For each stressor, cows were given a classification of severely, moderately or non-affected based on mobility scores, somatic cell counts and BCS change. The number of days from calving to onset of the first luteal phase was greater in cows that had one severe production stressor (median 44 days) or two moderate production stressors (41 days) compared with cows that had no stressors (31 days) (P=0.02 and P=0.04, respectively). More than one severe stressor increased the interval further. There was no difference between cows with one moderate stressor (median 38 days) and those with none (P=0.13). The delay to the first luteal phase was significantly longer in cows with two moderate stressors if the onset of one stressor occurred at the time when resumption of ovarian activity was expected. The presence of these production stressors in early lactation had no effect on the interval from calving to establishment of the next pregnancy or the number of inseminations required despite the negative effect on the onset of the luteal phase.

Reasons for Performing Study: Increased prevalence of obesity among UK horses and ponies demands evidence-based advice to promote weight loss., Hypothesis: Restriction of dry matter intake (DMI) to 1% of body mass (BM, 67% [corrected] of predicted maintenance digestible energy [DE] requirements) would promote weight loss without compromise to health., Methods: Five mature (mean ± s.e. 10 ± 2 years), overweight/obese pony mares (BM, 257 ± 20 kg: body condition score [BCS] 6.8/9 ± 0.5) were studied over 12 weeks. Animals were individually housed. Daily provision of a chaff-based, complete diet (measured DE, 8.5 MJ/kg DM) was restricted to 1% of actual BM as DMI daily. BCS, girth measurements and ultrasound-derived measures of subcutaneous fat depth overlying the gluteal region and 12th intercostal space (rib-eye) were recorded weekly. Body fat content was estimated at the beginning and end of the study by deuterium oxide dilution methods. Clinical biochemistry was monitored weekly. Behaviour was observed (24 h, 3/5 ponies) on 3 occasions., Results: BM decreased by 4.3 ± 1.1% during the first week and thereafter by 0.7 ± 0.1% of BM at end of Week 1 each week. BCS remained constant. Heart and belly girths, rump width and subcutaneous fat depth at rib-eye decreased significantly with time and BM. Fat comprised 45 ± 19% of BM loss. Fatter animals lost relatively more fat. With decreased feeding activity, time spent in 'play' and rest increased by 36 ± 11% and 438 ± 95%, respectively., Conclusions: This plane of nutrition resulted in an overall rate of weight loss of 1% of outset BM weekly. BCS was not a useful index of early weight loss but heart and belly girths and subcutaneous rib-eye fat were identified as alternative markers., Potential Relevance: This study provides an evidence-base for the management of weight loss in obese animals, especially those for which exercise may be contra-indicated., (© 2010 EVJ Ltd.)

Spherical (35 mm diameter) glass intra-uterine devices (IUDs) are commonly understood to modify sexual behaviour in performance mares. The effect of IUDs on the endocrinology (luteinising hormone, oestradiol, progesterone, testosterone), ovarian and uterine function, sexual and social behaviours associated with the ovarian cycle were evaluated during the natural breeding season in nine, nuliparous pony mares (4.6±0.5 years old, 230±12 kg bodyweight). Age and weight-matched groups were allocated to sham-treatment (n=5) or IUD-implantation (n=4) groups. Fifty complete ovarian cycles were evaluated, with 26 ovarian cycles monitored for the nine mares prior to treatment and a further 12 cycles were evaluated for mares in each of the groups post-treatment. Detailed characterisation of cyclic reproductive changes in the pony may be of value for controlled breeding programmes. The presence of an IUD was well-tolerated, but failed to modify any recorded aspect of reproductive physiology or behaviour., (Crown Copyright © 2009. Published by Elsevier Ltd. All rights reserved.)

Standard and modified measuring sticks were used to record height at the withers and a 'non-contact' laser was used to measure withers and loin heights. Sixty horses and ponies, ranging in height (115 to 155 cm) and body condition score (BCS) (moderate to obese) were measured by each method at 10-minute intervals for 40 minutes. Measurement series were repeated over three successive days. Unique regression models were constructed for method-specific data. Coefficients of variation were similar for stick and laser methods (0.002 to 0.004 per cent). Models were not influenced by day of measurement or BCS. Withers height decreased significantly (0.48 cm, 95 per cent confidence intervals -0.61 to -0.36 cm, P<0.001) over the first 20 minutes. In living animals, laser-derived measurements of withers height at T(20) exceeded stick measurements by approximately 1 cm (P<0.001). Loin height remained similar across time. Some alteration in relaxed withers height is an inevitable consequence of changes in muscle tone at the scapulothoracic synsarcosis.

Concentrations of gonadotrophins and prolactin were recorded in pony stallions castrated during the early breeding season, to examine the regulatory role of the gonad at a time when testosterone has been postulated to exert positive feedback on LH secretion. Further, gonadotrophin concentrations in geldings are reported to return to values within the normal range of the entire stallion. In an attempt to characterize this species-specific reversal, the gonadotrophin concentrations of 6 male ponies castrated on 25 March were monitored for 4 months, and 4 stallions were used to generate control data. Blood samples were collected daily, from 3 d before to 10 d after castration (Day 0), and weekly thereafter until Day 122. The pituitary response to castration was immediate. Castration resulted in a previously unreported, dramatic (13-fold) but transient (3 d) surge in circulating concentrations of LH. Concentrations of LH and FSH increased in a logarithmically scaled (LH, R2 = 0.77; FSH, R2 = 0.93) manner over the subsequent 5 wk, during which temporal changes in concentrations of both hormones were strongly correlated (R2 = 0.97). The ratio of plasma gonadotrophin concentrations was consistent throughout (LH:FSH, 1.43 +/- 0.04). Maximal concentrations of LH (20.58 +/- 1.97 ng/mL, Day 34.8 +/- 3.2) were attained approximately 2 wk before the peak in FSH (16.99 +/- 1.97 ng/mL, Day 49.7 +/- 3.0). Plasma gonadotrophin concentrations exceeded those of entire stallions throughout the study. The equine testes inhibited LH secretion during the early breeding season, and no chronic decrease in plasma gonadotrophin concentrations was recorded. However, the LH surge evident for 3 d immediately afer castration, may be related to the dynamic seasonal interaction between gonadal steroids and the regulation of pituitary gonadotrophin release.

Two comparative studies were undertaken using adult, female red and Père David's deer to examine the ovulatory response of these animals to a superovulation regimen and fertilization rates following inter- and intraspecific laparoscopic insemination. In Expt 1 six Père David's deer and 12 red deer hinds were treated during the breeding season with an intravaginal progesterone-impregnated controlled internal drug release device (CIDR) for 14 days, with 200 iu pregnant mares' serum gonadotrophin (PMSG) administered 72 h before the device was withdrawn and eight injections of ovine FSH given at 12 h intervals starting at the time of PMSG administration. Oestrous behaviour began one day after CIDR device withdrawal (Père David's deer: 24.00 +/- 2.32 h; red deer: 24.60 +/- 2.23 h). The duration of oestrus was greater in Père David's deer than in red deer (17.50 +/- 1.43 h and 8.25 +/- 3.25 h, respectively, P < 0.001). The peak LH surge of Père David's deer was 68.65 +/- 4.74 ng ml-1 occurring 29.00 +/- 2.41 h after removal of the CIDR devices. In comparison, the peak LH surge in red deer was 17.09 +/- 3.64 ng ml-1 (P < 0.001), occurring 24.00 +/- 0.00 h after CIDR device withdrawal. Pre-surge concentrations of LH were also greater (P < 0.001) in Père David's deer (1.37 +/- 0.11 ng ml-1) than in red deer hinds (0.41 +/- 0.02 ng ml-1).(ABSTRACT TRUNCATED AT 250 WORDS)

The oestrous cycles of fourteen red deer hinds (six yearling; eight more than 2 years old) were synchronized during the early breeding season by removal of a progesterone-containing intravaginal device and blood samples were taken at intervals of 3 h commencing 13 or 25 h later and continued for 54 h. The controlled internal drug release devices (CIDRs) were removed at 08:00 h (group 1; three yearlings and four adults) or 12 h later at 20:00 h (group 2; three yearlings and four adults). There was no significant effect of time of removal of CIDR on the interval to the onset of oestrus (group 1, 34.5 +/- 4.05 h; group 2, 42.14 +/- 7.8 h) on the time of peak concentration (group 1, 41.81 +/- 5.69 h; group 2, 41.71 +/- 7.81 h) or on duration of the luteinizing hormone (LH) surge (group 1, 15.00 +/- 0.95 h; group 2, 14.57 +/- 0.78 h). The six yearling animals exhibited oestrus and LH surge significantly later than the adults (55 +/- 4.2 versus 32 +/- 6.3 h for the LH surge for yearling and adult females, respectively). In a further experiment, 20 hinds were synchronized during the breeding season by removal of CIDR at two times of day 12 h apart and placed with a stag. Mating took place at a mean time of 42.1 +/- 2.4 h and 37.0 +/- 1.3 h later in the two groups. There was no significant effect of time of removal of CIDR upon time to onset of oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)

This study investigated the effect of melatonin treatment of ewe lambs on LH pulsatility in an attempt to examine the mechanism whereby melatonin advances the onset of puberty. Six ewe lambs were given intravaginal melatonin implants at 12.8 weeks of age. Another six lambs received empty implants. All lambs were serially blood sampled every 15 minutes for six hours on several occasions prior to the onset of puberty. One week after implantation LH pulse frequency and mean LH levels were higher in treated lambs than the control lambs (pulse frequency 0.13/h vs 0.03/h; mean LH levels 2.0 +/- 0.2 ng/ml vs 1.3 +/- 0.1 ng/ml; p less than 0.05). Melatonin treatment failed to alter pulse frequency after the initial increase. Puberty was advanced by 3 weeks in the treated group. In the second experiment six lambs received melatonin implants at 13 weeks of age and another six lambs served as control. In this experiment blood samples were taken intensively during the first few weeks after treatment. Results of this study show that mean plasma LH levels and LH pulse frequency were again higher during the first week after implantation. This transient increase in LH release may be part of the mechanism initiating the eventual advancement of puberty although the significance of this increase is questionable. In both experiments the LH response to estradiol injection was monitored at various times after treatment, but no effects of melatonin were found, although the magnitude of the response increased with age.

This study tested the role of melatonin in the regulation of seasonal physiological change in the pony stallion. Four 3-year-old, Welsh Mountain pony stallions were housed initially under the prevailing short-day photoperiod in December (8 of light [L]:16 h of darkness [D]) before being transferred to long days (16L:8D) on 13 January for the remaining 22 weeks of the study. On Day 76 (11 weeks later) the stallions began an 11-week period of daily melatonin treatment (20 mg orally, 8 h after lights on). Marked changes in mean plasma testosterone, beta-endorphin and cortisol concentrations occurred in response to long days and to subsequent melatonin treatment. Photostimulation produced a sharp rise in overall mean daily testosterone to a peak of 6.74 nmol/litre by Day 30. Values then fell to a nadir (3.17 nmol/litre) by Day 85, suggesting a role for melatonin in the termination of breeding activity in the horse. Cortisol and beta-endorphin values remained low throughout the first 11 weeks, but by Day 105 (Day 30 of melatonin treatment) concentrations had risen sharply, attaining a peak on Day 125 (510 pg beta-endorphin/ml, 50 ng cortisol/ml). Concentrations of both hormones had fallen by Day 77 of melatonin treatment (Day 152), perhaps as a result of refractoriness. Parallelism between beta-endorphin and cortisol suggests a pituitary origin for peripheral beta-endorphin. Diurnal variation in cortisol was observed under long days but no change in beta-endorphin was detected. Long days and melatonin treatment stimulated shedding of the winter and summer coats respectively, whereas growth rate was increased (2.03 kg/week) during the period of melatonin treatment relative to that of long days only (0.37 kg/week). The study provides evidence that the diurnal pattern of melatonin secretion mediates the reproductive and non-sexual responses to photoperiodic change in pony stallions.

The administration of melatonin via intravaginal sponges is an effective method of advancing the breeding season in ewes. In the present study the fertility of melatonin-treated ewes has been compared with that of ewes induced to ovulate by conventional treatment. On June 25, 15, 15-month-old ewes (group 1) were given intravaginal implants containing melatonin in silastic tubing. On August 8, 13 similar ewes (group 2) were given Veramix sponges which were removed 12 days later, when they were given 500 iu pregnant mare's serum gonadotrophin (PMSG). Two intact raddled rams were introduced to the combined groups on August 21. The mean date of mating was September 3 +/- 1.5 for group 1 and August 21 +/- 0.2 for group 2 ewes. All the ewes in group 1 and 10 in group 2 (77 per cent) were mated. All the ewes were slaughtered approximately 50 days after mating and their reproductive tracts removed. The mean ovulation rates were 2.1 and 2.3 in groups 1 and 2, respectively. The results indicate that conception rates of 87 per cent and 61.5 per cent of ewes put to the ram were obtained in the melatonin-treated and PMSG-treated groups, respectively. At slaughter the melatonin-treated group were found to have a mean of 1.47 live fetuses per ewe put to the ram and the PMSG-treated group a mean of 1.08. It can therefore be concluded that melatonin implantation is an effective method for the advancement of seasonal breeding in anoestrous sheep, and that the fertility achieved is at least as good as that given by conventional progestogen-PMSG treatment.