Macroscopic and Histological Morphology of the Equine Digital Cushion and Its Association with Subcutaneous Fat Thickness.

This study evaluated the influence of the Body Condition Score (BCS) and subcutaneous fat thickness on the tissue composition of the digital cushion in horses. Sixty mixed breeds of Criollo horses (21 males and 39 females) were sent for slaughter. All animals were submitted to BCS analysis, through visual antemortem evaluation, and then ultrasound evaluation to estimate the subcutaneous fat thickness. Macroscopic analyses of the thoracic and a pelvic limb (weight, volume, and density of the hooves and digital cushions) were performed. In addition, measurements of the area of internal structures to the hoof and histological analyzes were carried out to measure the areas of adipose, fibroelastic, and collagen fibers of the digital cushion. There were no macroscopic differences in the digital cushion between thoracic and pelvic limbs or between genders (P > .05). Likewise, the histological characteristics between the limbs and the genders were similar (P > .05). There was no correlation between the weight, volume, and density of the digital cushion with the BCS (P > .05). A positive correlation was observed with the area of adipose tissue (P = .038, R2 = 0.28) and a negative correlation with area of fibroelastic tissue (P = .005, R2 = -0.37) and collagen fibers (P = .003, R2 = -0.39). In conclusion, the adipose tissue, fibroelastic tissue, and collagen fibers of equine digital cushion alter their areas in the functions of the various subcutaneous fat patterns in horses.

Equine yolk sac: a stem cells source / Saco vitelino equino: una fuente de células madre

SUMMARY: Mesenchymal stem cells are characterized by in vitro high proliferation and multilineage potential maintenance. This study aimed to isolate and characterize equine YS mesenchymal stem cells and compare these with amniotic membranes. The yolk sac (YS) and amniotic membranes (AM) were obtained from 20 pregnant mares with gestational age around 30 days. Cells were cultured in α-MEM supplemented with 15 % FBS, 1 % antibiotic solution, 1 % L-glutamine and 1 % nonessential amino acids. To cell characterization we used cytogenetic analysis, fibroblast colony-forming unit assays, cell growth curves, immunophenotyping, flow cytometry, differentiation assays and teratoma formation. Results: Both cell sources presented fibroblastoid and epithelioid-like format. The YS cells have lower colony formation potential then AM ones, 3 versus 8 colonies per 103 plated cells. However, YS cells grew progressively while AM cells showed steady. Both, the YS and amnion cells immunolabeled for Oct-4, Nanog, SSEA-3, cytokeratin 18, PCNA, and vimentin. In addition, presented mesenchymal, hematopoietic, endothelial and pluripotency markers in flow cytometry. Discussion: Both cell sources presented high plasticity and differed into osteogenic, adipogenic, and chondrogenic lineages, and no tumor formation in nude mice was observed. The results suggest that horse YS may be useful for cell therapy such as amnion-derived cells.

RESUMEN: Las células madre mesenquimales se caracterizan por una alta proliferación in vitro y un mantenimiento potencial de múltiples líneas. Este estudio tuvo como objetivo aislar y caracterizar las células madre mesenquimales del saco vitelino equinas y compararlas con las membranas amnióticas. Se obtuvo el saco vitelino (SV) y las membranas amnióticas (MA) de 20 yeguas preñadas con edad gestacional de aproximadamente 30 días. Las células se cultivaron en α -MEM suplementado con 15 % de FBS, 1 % de solución antibiótica, 1 % de L-glutamina y 1 % de aminoácidos no esenciales. Para la caracterización celular utilizamos análisis citogenéticos, ensayos de unidades de colonias de fibroblastos, curvas de crecimiento celular, inmunofenotipaje, citometría de flujo, ensayos de diferenciación y formación de teratomas. Ambas fuentes celulares presentaron formato fibroblastoideo y epitelioide. Las células SV tienen un potencial de formación de colonias más bajo que las de MA, 3 versus 8 colonias por 103 células en placa. Sin embargo, las células SV crecieron progresivamente mientras que las células MA se mostraron estables. Tanto las células YS como las células amnios están inmunomarcadas para Oct-4, Nanog, SSEA-3, citoqueratina 18, PCNA y vimentina. Además, presentó marcadores mesenquimales, hematopoyéticos, endoteliales y pluripotenciales en citometría de flujo. Ambas fuentes celulares presentaron alta plasticidad y diferían en linajes osteogénicos, adipogénicos y condrogénicos, y no se observó formación de tumores en ratones. Los resultados sugieren que el SV de caballo puede ser útil para la terapia celular, como las células derivadas de amnios.

Impact of high-energy diets on the rumen environment and digital cushion in confined cattle / O impacto de dietas de alto valor energético sobre o ambiente ruminal e o tecido digital em bovinos confinados

This study aimed to identify gross and microscopic changes, caused by high-energy diets, in the rumen environment and hoof of confined beef cattle. The study sample comprised 40 confined heifers (Bos taurus) with no disease history divided into four experimental groups using different diets: Group 1 (D1, control), 48:52 forage:concentrate ratio; Group 2 (D2), 30:70 forage:concentrate ratio; Group 3 (D3), 30:70 forage:concentrate ratio + sucrose; Group 4 (D4), 100% concentrate. All animals underwent clinical examination, assessment of ruminal fluid pH and lameness, and sample collection after slaughter for histopathology of the hoof laminae and digital cushion and ruminal tissue. All dependent variables of the study were compared using the SPSS 20.0 statistical software. The variables that did not show normality (HR, RM, ST, and CRT) were compared with application of the Kruskal-Wallis test followed by the Dunnet's multiple comparison test. All other variables were submitted to analysis of variance (ANOVA) followed by Tukey's test. The different diets had an impact on the rumen environment (p<0.05) of the heifers assessed, with momentary general depression in the first 12 h after sucrose induction (D3), as well as mild clinical signs in D4. The animals in D3 and D4 presented lower motility (p<0.05) and ruminal pH (p<0.01) than those in D1. Of the 40 heifers, 27.5% (n=11) showed gross lesions in the epithelium of ruminal pillars, whereas 22.5% (n=9) of those in D3 and D4 presented these lesions. Sole corium thickness varied between heifers in D3 compared with those in D1 and D2 (p<0.05). Therefore, high-energy diets, as used in this study, alter some clinical parameters and the rumen environment, causing lesions in the rumen mucosa, and of lesser intensity, in the hoof corium and laminae, suggestive of laminitis.(AU)

O objetivo deste trabalho foi identificar alterações macroscópicas e microscópicas provocadas por dietas de alto valor energético no ambiente ruminal e casco de bovinos de corte (Bos taurus) em confinamento. Foram utilizadas 40 novilhas mantidas em confinamento, sendo divididas em 4 grupos experimentais: Grupo D1(controle) - 48% de volumoso e 52% de concentrado; Grupo D2 - 30% de volumoso e 70% de concentrado; Grupo D3 - 30% de volumoso e 70% de concentrado + sacarose; Grupo D4 - 100% de concentrado. Foi realizado exame clínico dos animais, avaliação do pH ruminal, da claudicação, além de coletas de amostras post-mortem para análises histopatológicas da região laminar do casco, coxim digital e do rúmen. Todas as variáveis dependentes do estudo foram comparadas por meio do software estatístico SPSS 20.0. As variáveis sem normalidade (FC, MR, Tc e TPC) foram comparadas pelo teste de Kruskal Wallis, seguido pelo teste de comparação múltipla de Dunnet. Todas demais foram submetidas a análise de variância (ANOVA), seguida pelo teste de Tukey. Foi observado impacto das distintas dietas ao ambiente ruminal (p<0,05). Os grupos D3 e D4 exibiram menor motilidade (p<0,05) e pH ruminal (p<0,01). 27,5% (n=11) dos animais tiveram registros macroscópicos de lesões no epitélio dos pilares ruminais enquanto que os submetidos às dietas 3 e 4 foi observada ocorrência de 22,5% (n=9) de lesões macroscópicas no rúmen. As espessuras do córium solear variaram entre os grupos que receberam a indução de sacarose em relação ao grupo controle e novilhas alimentadas com a dieta 2 (p<0,05). Sendo assim dietas com elevada densidade energética, na condição em que foi empregada, alteraram alguns parâmetros clínicos e o ambiente ruminal, provocando lesões na mucosa do rúmen e de pouca intensidade no córium e tecido laminar, sugestiva de laminite.(AU)