The horse is a non-ruminant herbivore adapted to eating plant-fibre or forage-based diets. Some horses are stabled for most or the majority of the day with limited or no access to fresh pasture and are fed preserved forage typically as hay or haylage and sometimes silage. This raises questions with respect to the quality and suitability of these preserved forages (considering production, nutritional content, digestibility as well as hygiene) and required quantities. Especially for performance horses, forage is often replaced with energy dense feedstuffs which can result in a reduction in the proportion of the diet that is forage based. This may adversely affect the health, welfare, behaviour and even performance of the horse. In the past 20 years a large body of research work has contributed to a better and deeper understanding of equine forage needs and the physiological and behavioural consequences if these are not met. Recent nutrient requirement systems have incorporated some, but not all, of this new knowledge into their recommendations. This review paper amalgamates recommendations based on the latest understanding in forage feeding for horses, defining forage types and preservation methods, hygienic quality, feed intake behaviour, typical nutrient composition, digestion and digestibility as well as health and performance implications. Based on this, consensual applied recommendations for feeding preserved forages are provided.

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‐ (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.

Nuchal crest adiposity in horses and ponies has been associated with an enhanced risk of metabolic health problems. However, there is no current information on the prevalence of, and risk factors specific to, nuchal crest adiposity in horses and ponies. In addition, the cresty neck score has not previously been utilised across different seasons within a UK leisure population, it is not know whether nuchal crest adiposity shows the same seasonal trends as general obesity. A Cresty Neck Score (CNS, 0–5) was given to 96 horses with access to pasture (>6 h per day) at the end of winter and at the end of summer in order to obtain two prevalence estimates. Risk factors were assessed using the single outcome cresty neck/no cresty neck in either season (binary), from owner questionnaires and analysed using a mixed effects logistic regression model (outcome variable CNS <3 or CNS ≥3/5). Agreement between winter and summer scores was assessed using weighted Kappa methods. Winter CNS values were significantly higher than summer CNS values (p = 0.002) indicating a systematic bias. The prevalence of a CNS ≥ 3/5 was 45.83% at the end of winter, falling to 33.33% at the end of summer and was higher in ponies (<14.2 hh) than horses (≥14.2 hh) in both seasons. This may reflect a real winter increase in regional fat deposition, or an increased difficulty in obtaining an accurate estimate of regional adiposity in winter months. Breed was the strongest risk factor for CNS ≥3/5 in both seasons, with native UK breeds appearing to be most at risk (p < 0.001). In a separate, small validation study, the CNS showed good inter-observer reliability. The prevalence of a CNS ≥3/5 was higher at the end of winter than at the end of summer, which was the opposite pattern seasonal variation to that observed for general obesity. Further studies are required to investigate the potential influence of time of year upon CNS interpretation and studies utilising the CNS should consider potential seasonal variability in nuchal crest adiposity.

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 ofFirmicutesandTenericutesand 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 withSphaerochaeta (Spirochaetesphylum) 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.

Introduction: Arthritic diseases are characterized by the degradation of collagenous and noncollagenous extracellular matrix (ECM) components in articular cartilage. The increased expression and activity of matrix metalloproteinases (MMPs) is partly responsible for cartilage degradation. This study used proteomics to identify inflammatory proteins and catabolic enzymes released in a serum-free explant model of articular cartilage stimulated with the pro-inflammatory cytokine interleukin 1β (IL-1β). Western blotting was used to quantify the release of selected proteins in the presence or absence of the cyclooxygenase-2 specific nonsteroidal pro-inflammatory drug carprofen. Methods: Cartilage explant cultures were established by using metacarpophalangeal joints from horses euthanized for purposes other than research. Samples were treated as follows: no treatment (control), IL-1β (10 ng/ml), carprofen (100 μg/ml), and carprofen (100 μg/ml) + IL-1β (10 ng/ml). Explants were incubated (37°C, 5% CO₂) over twelve day time courses. High-throughput nano liquid chromatography/mass spectrometry/mass spectrometry uncovered candidate proteins for quantitative western blot analysis. Proteoglycan loss was assessed by using the dimethylmethylene blue (DMMB) assay, which measures the release of sulfated glycosaminoglycans (GAGs). Results: Mass spectrometry identified MMP-1, -3, -13, and the ECM constituents thrombospondin-1 (TSP-1) and fibronectin-1 (FN1). IL-1β stimulation increased the release of all three MMPs. IL-1β also stimulated the fragmentation of FN1 and increased chondrocyte cell death (as assessed by β-actin release). Addition of carprofen significantly decreased MMP release and the appearance of a 60 kDa fragment of FN1 without causing any detectable cytotoxicity to chondrocytes. DMMB assays suggested that carprofen initially inhibited IL-1β-induced GAG release, but this effect was transient. Overall, during the two time courses, GAG release was 58.67% ± 10.91% (SD) for IL-1β versus 52.91% ± 9.35% (SD) with carprofen + IL-1β. Conclusions: Carprofen exhibits beneficial anti-inflammatory and anti-catabolic effects in vitro without causing any detectable cytotoxicity. Combining proteomics with this explant model provides a sensitive screening system for anti-inflammatory compounds.

In humans, adipose tissue (AT) originating from different depots shows varying gene expression profiles. In horses, the risk of certain metabolic disorders may also be influenced by the impact of specific AT depots. Macrophage infiltration in human and rat AT is considered to be a source of inflammatory changes. In horses, this relationship has not been extensively studied yet. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), a useful method to evaluate differences in mRNA expression across different tissues, can be used to evaluate differences between equine AT depots. For a correct interpretation of the RT-qPCR results, expression data have to be normalized by the use of validated reference genes. The main objectives of this study were to compare mRNA expression of inflammation-related genes, as well as adipocyte morphology and number between different equine AT depots; and in addition, to investigate the presence of antigen presenting cells in equine AT and any potential relationship with adipokine mRNA expression.