Assessment of apoptosis in epidermal lamellar cells in clinically normal horses and those with laminitis

Objective - To determine and compare the number, type, location, and distribution of apoptotic epidermal cells in the laminae of clinically normal horses and horses with laminitis.Sample Population - Formalin-fixed samples of digital lamellar tissue from 47 horses (including clinically normal horses [controls; n = 7], horses with acurte [4] and chronic [7] naturally acquired laminitis, and horses with black walnut extract-induced [11] or carbohydrate overload-induced [18] laminitis).Procedure - Blocks of paraffin-embedded lamellar tissues were stained for DNA fragmentation with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Differential immunohistochemical staining for caspases 3 and 14 were used to confirm apoptosis.Results - the number of TUNEL-positive epidermal cells per 0.1 mm of primary laminae was significantly greater in the acute laminitis group than in the other groups. In the acute laminitis group, there were 17 and 1,025 times as many TUN EL-positive basal layer cells and keratinocytes, respectively, compared with the control group. Apoptosis of TUNEL-positive basal layer cells was confirmed by results of caspase 3 immunohistochemical staining. The TUNEL-positive keratinocytes did not stain for caspases 3 or 14.Conclusions and Clinical Relevance - the large number of apoptotic basal layer cells detected in the lamellar tissue of horses with acute naturally acquired laminitis suggests that apoptosis may be important in the development of acute laminitis. The role of the large number of TUNEL-positive keratinocytes detected in the interface of primary and secondary epidermal laminae of horses with acute laminitis remains to be elucidated.

Changes in plasma protein concentrations in ponies with experimentally induced alimentary laminitis

Objective-To determine whether plasma protein concentrations were altered in ponies with alimentary laminitis.Animals-12 adult ponies.Procedure-Acute laminitis was induced in 6 ponies by oral administration of carbohydrate (85% corn starch, 15% wood flour); the other 6 ponies were used as controls. A physical examination was performed and blood samples were collected immediately before and 4, 8, 12, 24, and 28 hours after administration of carbohydrate. Plasma protein concentrations were determined by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis.Results-19 plasma proteins ranging from a molecular weight of 24,000 to a molecular weight of 350,000 were identified in all 12 ponies. Plasma concentrations of proteins with molecular weights of 350,000 (fibrinogen), 130,000 (ceruloplasmin), 118,000 (c-reactive protein), 67,000 (alpha(1)-antitrypsin I), 65,000 (alpha(1)-antitrypsin II), 50,000 (haptoglobulin), and 45,000 (acid glycoprotein) were significantly increased in ponies with laminitis, compared with concentrations in control ponies.Conclusion-Changes in plasma protein concentrations are detectable within 4 hours after the onset of alimentary laminitis in ponies.Clinical Relevance-Measurement of plasma protein concentrations may be useful in monitoring the progression of laminitis in ponies.

Morphological abnormalities and apoptosis in lamellar tissue of equines after intestinal obstruction and treatment with hydrocortisone

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da heparina sódica na ocorrência de laminite eqüina induzida por sobrecarga de carboidratos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Alterações morfológicas de tecido laminar do casco e parâmetros clínicos e laboratoriais de equinos com síndrome cólica letal

As afecções gastrintestinais dos cavalos são agravadas por complicações como a laminite, cuja etiopatogenia está relacionada à degradação da membrana basal do tecido laminar por metaloproteinases (MMPs). A ativação das MMPs pode ocorrer devido à liberação local de citocinas inflamatórias ou enzimas provenientes de leucócitos infiltrados no tecido laminar. O objetivo deste trabalho foi avaliar as alterações morfológicas do tecido laminar de equinos com síndrome cólica letal e sua provável associação com parâmetros clínicos e laboratoriais. Observou-se intensa destruição da arquitetura laminar, principalmente nos animais com alterações físicas e laboratoriais mais acentuadas, como tempo de preenchimento capilar prolongado (TPC), membranas mucosas congestas, taquicardia, hemoconcentração e redução nas contagens de plaquetas e leucócitos. Os resultados sinalizam o provável momento do desenvolvimento de lesões do tecido laminar em equinos com síndrome cólica, no qual é possível adotar medidas preventivas contra a laminite.

Lipocalina associada à gelatinase de neutrófilos (NGAL) e calprotectina no tecido laminar de equinos após obstrução jejunal, tratados ou não com hidrocortisona

A laminite é uma doença podal grave que acomete os equídeos, sendo responsável por intenso sofrimento. Neste estudo foram pesquisadas a presença de calprotectina por meio da imunoistoquímica, e de lipocalina associada à gelatinase de neutrófilos (NGAL), por zimografia, no tecido laminar do casco de equinos após obstrução intestinal. Os animais foram divididos em quatro grupos: Grupo controle (Gc), contendo sete animais normais, sem procedimento cirúrgico; Grupo Instrumentado (Gi), contendo cinco animais, os quais passaram por todo o procedimento cirúrgico sem sofrerem obstrução intestinal; Grupo Não Tratado (Gnt), contendo quatro equinos submetidos a obstrução intestinal do jejuno por distensão de balão intraluminal, sem tratamento; e Grupo Tratado (Gt), contendo quatro equinos submetidos a obstrução intestinal, e tratados preventivamente com hidrocortisona. Houve imunomarcação de calprotectina em todos os grupos experimentais, com aumento nos equinos do grupo distendido em relação ao Gc. Com relação ao NGAL, houve aumento também do Gnt e do Gi em relação ao Gc. O Gt não diferiu dos demais. Conclui-se que a distensão do intestino delgado pode promover acúmulos de leucócitos nos cascos de equinos e que o NGAL é um método viável para se detectar infiltração neutrofílica em equinos. Novos estudos deverão ser realizados para se verificar possível benefício anti-inflamatório da hidrocortisona no casco de equinos com obstrução intestinal.

Acidosis ruminal en bovinos lecheros: Implicaciones sobre la producción y la salud animal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Alterações morfológicas, apoptose e expressão de metaloproteinases no tecido laminar de equinos submetidos à obstrução intestinal experimental: efeitos da hidrocortisona

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expressão das metaloproteinases e morfologia do tecido laminar do casco de equinos submetidos à obstrução intra-luminal do cólon menor

Pós-graduação em Medicina Veterinária - FCAV

Tamponamento cecal: aspectos clínico, fisiopatológico e terapêutico na laminite experimental em eqüinos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Laminite experimental: aspectos morfológicos, morfométricos e ultra-estruturais das lâminas dérmicas e epidérmicas do casco de eqüinos tratados com a trinitroglicerina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo das correlações entre lesões ósseas podais diagnosticadas pela radiografia com o peso vivo, circunferência torácica, D.M.O.(determinada por densitometria óptica radiográfica)concentrações séricas de Ca, P, Mg, Zn, Fosfatase Alcalina, Proteínas Plasmáticas totais, Globulina, Creatina quinase, GamaGlutamiltransferase, Albumina, Aspartato-aminotransferase, Osteocalcina, pH do Rúmen e Hemograma em bovinos fêmeas da raça Nelore, sem sintomas clínicos de doença podal

Pós-graduação em Medicina Veterinária - FMVZ

Avaliação das características do líquido ruminal, hemogasometria, atividade pedométrica e diagnóstico de laminite subclínica em vacas leiteiras

The objectives of this study were to evaluate the characteristics of ruminal fluid, blood gas analysis, pedometer activity and suclinical laminitis occurrence, through the presence of secondary foot diseases in high production dairy cows, from a commercial herd. 200 Holstein cows originating from the same farm, located in Araçatuba/SP, Brazil, were divided into four groups, which are established from the daily milk production. Initially clinical examination of the cows was procedure, followed by sampling of rumen fluid, by esophageal tube. Fluid was evaluated for pH, color, odor, consistency, sedimentation, flotation and methylene blue reduction test. Venous blood samples were also collected for blood gas analysis, in addition to collecting data from pedometrics (number of steps) and daily milk production. Data were tabulated and submitted to correlation analysis. No animal had reported changes in rumen pH. Non-acid base imbalance were found, since the values of blood pH, PCO2, TCO2, HCO3- and BE were normal during hemogasimetric analysis. The pedometric was effective as a screening method to cows with foot diseases. It demonstrated reduction in the number of steps due to pain, correlated with lower milk production. However, the identification of foot lesions was only possible through specific digital clinical examination. The occurrence of 49.5 % of herd foot problems was due to the risk factors present in the farm. The abrasive concrete and inadequate facilities, also associated with the possible occurrence of sub-acute ruminal acidosis, were observed as risk factors. However, sub-acute ruminal acidosis was undiagnosed by the methods used. The correlation between the values of ruminal pH, and blood gas analysis pedometrics showed efficient for the early diagnosis of foot diseases and also in establishing the etiology of these diseases. Subclinical laminitis occurred primarily in cow's herd, considering the multifactorial etiology of this disease, occurrence and distribution of foot diseases diagnosed.

Atualidades no tratamento da laminite em equinos

Laminitis is still a major cause of lameness in horses, causing damage to many interested in it. It has been long studied, but there are several points that are not yet fully understood. Doubts about its pathogenesis end up extending at the treatments. Just as there are several theories that attempt to explain the mechanisms of its development, also appear treatments that are sometimes contradictory and inconsistent. Mostly times, laminitis is a consequence of another disease process that occurs systemically in the body. Among them are the main grain overload and processes that cause the animal to the table of endotoxemia, strangled colic, metritis with retained placenta and pleuropneumonia. The proposed treatments include the use of anti-inflammatory drugs, cryotherapy, use of vasodilators and correction of Starling forces, and other measures to support the frog. The stall should be bedded deeply with sand or other material that provides support to the frog

Respostas inflamatórias sistêmica, do casco e do cólon menor submetido à distensão em equinos

Pós-graduação em Medicina Veterinária - FCAV