A cross-sectional study of geriatric horses in the United Kingdom. Part 1: Demographics and management practices

Reasons for performing study: With growing numbers of aged horses, geriatric medicine is becoming increasingly important in equine veterinary practice; however, there is a paucity of information on the UK equine geriatric population. Objectives: To describe the demographic characteristics of the equine geriatric population and to assess management practices undertaken by owners of geriatric horses (aged ≥15 years). Methods: A cross‐sectional study was conducted, surveying a randomly selected sample of veterinary registered owners with horses aged ≥15 years, using a self‐administered postal questionnaire. Results: Horses aged ≥15 years represented 29% of the equine population in the study area. The response rate to the questionnaire survey was 80.2%. Management practices were similar to those previously described for the general equine population. However, the level of exercise and feeding practices changed significantly with increasing age. A summary of the demographic characteristics, feeding and management of horses in a sample of the geriatric equine population is presented. Conclusions and potential relevance: The management of geriatric horses has a direct effect on their health and welfare and this study has provided valuable information on the demographics and management of geriatric horses and ponies in the UK. The information can be used to form targeted owner education programmes for geriatric health and should aid in the investigation and prevention of disease.     

Ethyl pyruvate decreases proinflammatory gene expression in lipopolysaccharide-stimulated equine monocytes

Monocytes are among the initial cells that interact with circulating LPS. Binding of LPS to monocyte surface receptors triggers an intracellular signaling cascade and results in the production of proinflammatory cytokines. Ethyl pyruvate, a stable derivative of pyruvate, has been effective in mitigating LPS induced alterations in isolated human monocytes. We hypothesized that ethyl pyruvate would suppress proinflammatory gene expression in LPS-stimulated equine monocytes without affecting cell viability. Equine monocytes were isolated from whole blood using a sediment-gradient centrifugation protocol and enriched to 76% purity by adhesion to tissue culture dishes. Isolated monocytes were incubated with 0, 1, 5, 10 and 50 mM ethyl pyruvate. Cell viability, production of caspase 3/7, and caspase-3 gene expression were determined. In a separate experiment, monocytes were stimulated with LPS (0.1 ng/ml for 1h) followed by incubation with 0, 1, 5, or 10 mM ethyl pyruvate for 1 h. Proinflammatory gene expression was determined by real-time PCR. Ethyl pyruvate at 50 mM adversely affected monocyte viability. Ethyl pyruvate at 10mM or less had no significant effect on monocyte viability, and did not increase activity of caspase 3/7 nor caspase-3 gene expression. Incubation with LPS alone induced a significant upregulation in proinflammatory gene expression. Subsequent treatment of monocytes with ethyl pyruvate significantly reduced IL-8 expression in LPS stimulated monocytes at 5 mM, and IL-8, TNF-α and COX-2 at 10 mM. No beneficial effect on expression of IL-1β or IL-6 was detected. Overall, 10 mM ethyl pyruvate did not adversely affect monocyte viability and suppressed LPS-induced proinflammatory gene expression. Ethyl pyruvate may be a beneficial anti-inflammatory therapy in equine endotoxemia.     

Further insights into the characterization of equine adipose tissue-derived mesenchymal stem cells

Adipose tissue-derived stem cells (ADSCs) represent a promising subpopulation of adult stem cells for tissue engineering applications in veterinary medicine. In this study we focused on the morphological and molecular biological properties of the ADSCs. The expression of stem cell markers Oct4, Nanog and the surface markers CD90 and CD105 were detected using RT-PCR. ADSCs showed a proliferative potential and were capable of adipogenic and osteogenic differentiation. Expression of Alkaline phosphatase (AP), phosphoprotein (SPP1), Runx2 and osteocalcin (OC) mRNA were positive in osteogenic lineages and peroxisome proliferator activated receptor (Pparγ2) mRNA was positive in adipogenic lineages. ADSCs show stem cell and surface marker profiles and differentiation characteristics that are similar to but distinct from other adult stem cells, such as bone marrow-derived mesenchymal stem cells (BM-MSCs). The availability of an easily accessible and reproducible cell source may greatly facilitate the development of stem cell based tissue engineering and therapies for regenerative equine medicine.     

Presence of bluetongue virus in the marginal zone of the spleen in acute infected sheep

Bluetongue virus, a member of the genus Orbivirus of the family Reoviridae, is the causative agent of bluetongue, which is a non-contagious Culicoides mediated blood-borne disease. The present study characterizes the pathogenicity of a Taiwan prototype BTV2/KM/2003 in Corriedale sheep inoculated subcutaneously into the ear pinna. Histologically, multifocal petechiated hemorrhage, with mild to moderate inflammation and edema, were present in the contralateral ear pinna, tongue, and facial skin, without remarkable lesions in lymphoid organs. By days post-infection (DPI) 7, viral VP7 antigen, detected by immunohistochemistry, presented in the spleen, chiefly located in the outer rim of <3 cell thickness of marginal zone macrophages bordering the marginal zone and red pulp, and T lymphocytes of the red pulp. By DPI 11, viral signals shifted from the marginal zone to macrophages and small lymphocytes within follicles of the spleen. In situ hybridization with VP7 gene probe detected strong signals in the spleen, chiefly spanning the whole width of 5-10 cell thickness of the marginal zone, including the marginal zone macrophages and marginal zone B cells, as well as macrophages of sheathed capillaries in the red pulp. This study demonstrates molecular as well as morphologic evidence of the presence of bluetongue virus in the marginal zone of the spleen, most likely associated with viremia in acute infection, as previously demonstrated by the authors.     

Investigation of endothelial cells as an in vivo nidus of Anaplasma marginale infection in cattle

Continuous culture of Anaplasma marginale in endothelial cells and the potential implications for vaccine development heightened interest in determining the importance of endothelial cells in the A. marginale life cycle. A. marginale-infection trials were performed to determine if endothelial cells are an in vivo host cell in cattle and if A. marginale from in vitro endothelial cells were infective to cattle. Adult, immunocompetent steers were infected by tick-feeding transmission and were euthanized at different points in the parasitemic cycle. Based on quantitative PCR, the tissue distribution of A. marginale DNA during peak and trough parasitemia was variable with higher quantities observed in spleen, lung, hemal nodes, and abomasum. A. marginale was not conclusively identified in tissue endothelial cells from the steers' tick-bitten dermis or post-mortem tissues using three microscopy techniques (dual indirect immunofluorescence, transmission electron microscopy, and in situ DNA target-primed rolling-circle amplification of a padlock probe). Intravenous inoculation of spleen-intact or splenectomized calves with endothelial cell culture-derived VA isolate A. marginale did not cause seroconversion or clinical anaplasmosis regardless of whether the endothelial culture-derived bacteria were inoculated as host cell-free organisms or within endothelial cells and regardless of the type of endothelial cell culture used - RF/6A primate endothelial cells or primary bovine testicular vein endothelial cells. Data presented here suggest that endothelial cells are likely not a pivotal component of the A. marginale life cycle in vivo.     

Effects of dietary energy concentration, nonstarch polysaccharide concentration, and particle sizes of nonstarch polysaccharides on digesta mean retention time and gut development in laying hens

1. From an experiment with 504 laying hens (ISA Brown strain, 18–40 weeks of age), 90 40-week old hens were used for determining digesta mean retention time (MRT) and gut weight development. This experiment comprised 6 dietary treatments according to a 2?×?3 factorial design. Factors were dietary apparent metabolisable energy (AME) concentration (11·8 vs 10·6?MJ/kg), insoluble nonstarch polysaccharides (NSP) concentration (65 vs 134?g/kg), and fine vs coarse particle sizes of added NSP. Titanium recovery in different gut segments was used as an indicator of MRT.

2. Increasing NSP concentration prolonged MRT in the crop (68 vs 34?min) and total foregut (91 vs 57?min) compared with control NSP. Reducing energy concentration prolonged MRT in the colon (26 vs 7?min), and total hind gut (30 vs 9?min), compared with control energy. Overall MRT was not affected by dietary treatments.

3. Increasing NSP concentration increased relative weights of the empty proventriculus-gizzard and its contents by 30% (25·2 vs 19·4?g/kg) and 18% (15·4 vs 13·0?g/kg), respectively, compared with control NSP diets.

4. MRT in the foregut was prolonged as daily insoluble NSP intake increased, and this was more pronounced in hens given coarsely ground NSP, compared with finely ground. A prolonged MRT in the foregut seemed to indicate a higher level of satiety, which may contribute to a lower feather pecking pressure in laying hens.