Glucose is a fundamental energy source, the utilization and regulation of which impacts exercise performance and health. These studies have used modeling techniques to evaluate glucose kinetics and dynamics in equids and developed tests to evaluate the status of glucose metabolism. In Part I, 12 exercise-trained Arabians underwent insulin-modified FSIGT (with minimal model analysis) and single-injection glucose tracer (with compartmental analysis) studies at rest and during exercise to evaluate the effects of exercise on glucose kinetics and dynamics. These geldings were maintained on pasture, but adapted for 4 months to twice-daily feeding of feeds rich in sugar and starch (SS, n=6: NSC 45%, Fat 3%, NDF 24%) or fat and fiber (FF, n=6:NSC 13%, Fat 11%, NDF 45% ). Exercise increased insulin sensitivity (P = 0.070) and glucose transport (P < 0.038). Although variables were not different between FF and SS horses at rest, during exercise SS horses tended to have lower (P = 0.085) insulin sensitivity and increased (P = 0.043) glucose utilization compared to FF horses. In Part II, satisfactory proxies for minimal model parameters were developed to facilitate the evaluation of insulin sensitivity in larger populations. These proxies were applied to a population of 163 ponies and used to characterize metabolic differences between ponies predisposed to pasture laminitis (PL) from ponies not predisposed (NL). A subset of 14 ponies (7 PL, 7 NL) also underwent the FSIGT for minimal model analysis. Ponies predisposed to laminitis were found to have lower insulin sensitivity (P < 0.007) and higher insulin secretory response (P < 0.045) by both the minimal model and proxies, and higher (P < 0.001) circulating triglycerides and body condition score. Cut-point analysis for these variables was used to define a pre-laminitic metabolic syndrome with total predictive power of 78% to identifity ponies at risk for developing pasture laminitis. Increased insulin resistance and prevalence of lamintis were associated with increased non-structural carbohydrates in spring pasture. These studies demonstrate the importance of glucose regulation for exercise and animal health. When glucose regulation is altered in the case of insulin resistance, performance could be impacted and diseases such as laminitis may occur. Insulin resistance may be moderated by exercise or by avoiding sugar and starch in feeds and pasture. / Ph. D.

The objective of this study was to investigate whether it is possible to create acute laminitis in young ruminating calves by feeding high carbohydrate diets. Three experiments were performed. In the first, 16 calves, 17 wk of age, were fed one of four diets (4 replications) that contained either 71 or 81% of TDN and either 15 or 20% CP. Jugular blood and rumen fluid were sampled and hoof temperature measured at frequent intervals over a subsequent 2-d period. Calves responded acutely to the 81% TDN diets by anorexia, stiffness and diarrhea. Ruminal pH was lower and L- and D-lactates greater in the rumen of 81% TDN treatments. Total ruminal VFA decreased as pH declined. Proportion of acetate increased while propionate decreased in 81% TDN treatments. Butyrate differed but was not dietary related. Whole blood L-lactate did not differ by treatments. Blood D-lactate increased significantly in the calves fed 81% TDN, peaking at 32 h (65 mg/dL). Hoof temperature was significantly lower in 81% TDN treatments at 28-32 h. In the second experiment dietary sodium bicarbonate (.9% of DM) attenuated lactic acidosis in animals which consumed high quantities of concentrate. Although some animals in the buffer group suffered from acidosis, sickness was potentiated in nonbuffer group. Buffer inhibited the decline of ruminal fluid pH, and the increase of lactate in the rumen. Total VFA in the rumen declined with pH. Proportions of major VFA remained unchanged. Blood L-lactate increased at 28 h in animals which did not receive buffer. Blood D-lactate increased in both treatments and was greater in nonbuffer treatment. In the third experiment, 24 calves, 17 wk of age, were fed diets containing either 68 or 80% TDN. The latter diet was supplemented with either ionophore, buffer or contained no supplementation. Sudden introduction of the diets resulted in transient lactic acidosis. Buffer was more efficient in the prevention of acidosis than ionophore. The acetate : propionate (Ac:Pr) ratio tended to be lower in ionophore treatment. Growth performance did not differ between the treatments. Acute laminitis was not detected but the reduction of ruminal pH and a many fold increase in blood D-lactate observed in this study, may contribute to occurrence of laminitis. / Ph. D.

Submitted by Ubirajara Cruz (ubirajara.cruz@gmail.com) on 2017-05-10T11:33:05Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Fernanda_Pazinato.pdf: 1717141 bytes, checksum: 85ea4dd2cdaca042f7c8c423f2f8d128 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-05-10T13:42:16Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Fernanda_Pazinato.pdf: 1717141 bytes, checksum: 85ea4dd2cdaca042f7c8c423f2f8d128 (MD5) / Made available in DSpace on 2017-05-10T13:42:16Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Fernanda_Pazinato.pdf: 1717141 bytes, checksum: 85ea4dd2cdaca042f7c8c423f2f8d128 (MD5) Previous issue date: 2015-04-02 / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul - FAPERGS / Foram realizados dois estudos, com o objetivo de relacionar os aspectos anatomopatológicos da placenta de éguas Puro Sangue Inglês, com os dados da égua e peso do potro. O primeiro estudo descreve os achados histopatológicos das placentas de éguas PSI saudáveis e sua relação com a macroscopia placentária e dados das éguas. Foram utilizadas 188 éguas, cujas placentas foram avaliadas macro e microscopicamente As lâminas foram coradas por hematoxilina-eosina e ácido periódico de Schiff (PAS). Os dados avaliados nas éguas foram idade, número de partos, tempo de gestação, tempo de eliminação da placenta, comprimento do cordão umbilical, peso da placenta, e apresentação de sinais clínicos de placentite na gestação avaliada. Na macroscopia 79 placentas não demonstraram alteração, 87 apresentaram artefatos e 22 tinham placentite. Na histologia, observou-se que em 30 placentas o epitélio coriônico apresentava mais vacúolos, os quais se coravam pelo PAS. As éguas com esse tipo de placenta eram mais velhas e com maior número de partos. Observou-se 40 casos de placentites, sendo 19 crônicas e 21 agudas. Em 19 casos verificou-se hipoplasia de microcotilédones. Em 93% dos casos, os achados macroscópicos foram consistentes com a histologia placentária. A presença de sinais clínicos foi condizente com a histologia de placentite, entretanto, 19,3% das éguas com placentite na histologia, não demonstraram sinais clínicos. No segundo estudo avaliou-se a morfometria vascular da placenta de éguas PSI com laminite crônica. Foram utilizadas 26 éguas, 13 com apresentação de laminite crônica na gestação avaliada, e 13 éguas saudáveis. A pressão arterial sistólica e frequência cardíaca foram obtidas de 10 éguas, de cada grupo. As placentas eram avaliadas macroscopicamente e coletados fragmentos para histopatologia e morfometria. Foram obtidas imagens digitalizadas de corpo uterino, corno gravídico e corno não gravídico, e as medidas realizadas através do software NIH ImageJ 1.48r. As medidas dos vasos foram diâmetro vascular total, diâmetro de luz vascular e espessura de parede vascular. As medidas dos microcotilédones foram área total de microcotilédones, área capilar de microcotilédones e diferença entre área total de microcotilédones e capilar. Foram considerados idade da égua, tempo de gestação, número de partos, peso da placenta e peso do potro. As éguas com laminite crônica demonstraram quadro de hipertensão, com menor tempo gestacional, peso da placenta e do potro. Na morfometria estas demonstraram menor luz vascular, maior espessamento de parece vascular e menor área capilar nos microcotilédones. A laminite cursa com quadro hipertensivo, sendo este possivelmente responsável por alterações morfométricas vasculares. / Were made two studies, attempt to the relation between placental anatomopathologyc and data from Thoroughbreds mares and foal weight. The first study aimed to describe the histopathological features of the placentas of healthy Thoroughbred mares at foaling and to evaluate their relationship with the gross placental and mare’s data. The study was performed using 188 Thoroughbred mares, whose the placentas were grossly and histologic evaluated. The slides were stained using hematoxylin and eosin and periodic-acid-Schiff (PAS) reagents. The data from mares were age, number of parturition, gestational age, placental elimination time, umbilical-cord length and placental weight and signs of placentites. In grossly evaluation were observed 79 placentas without alterations, 87 with artifactuals findings and 22 placentites. In histology were observed 30 placentas with intense cytoplasmic vacuolization of the epithelial areolar cells, that stained by PAS. These mares were older and larger parturition number. In 40 of the placentas, the histopathological changes consisted of placentitis, 19 these with chronic placentites and 21 acute placentites. Also observed 19 placentas with hypoplasia/atrophy from microcotyledones. In 93% of cases the placental grossly were consistently with histologic findings. The mares that demonstrate clinical sings of placentitis showed correspondent histologic findings, however in 19,3% of mares have placentites in histology without presence of clinical sings. On second study the aim was evaluate the placental microcotyledonary and vascular morphometry, in Thoroughbreds mares with chronic laminitis. Twenty six mares divided in two groups were used, 13 these with chronic laminitis, and 13 mares healthy. Arterial systolic pressure and cardiac frequency were obtained in 30 days prefoaling. The placentas were submit to grossly evaluation and taken samples to histopathological and morphometric study. Digital images analysis was made from placental uterine body, gravid horn and non gravid horn, and measures by software ImageJ 1.48r. Measures of allantoic surface vessels was total vessel diameter, lumen vessel diameter and wall vessel thickness. In microcotyledons the measures was total area from microcotyledons, capillary area from microcotyledons, difference capillary/total area from microcotyledons and relation between these areas. The age of mare, gestational age, number of parturition, placental weight and foal weight were considered. The mares with chronic laminitis showed hypertension, lower gestational period, placental and foal weight. These mares showed lower lumen vessel diameter, thickness of wall vessel e less capillary area from microcotyledons.

In horses, a painful and often debilitating disease known as laminitis can result in impaired function and, in severe cases, euthanasia. Equine Metabolic Syndrome (EMS) is a syndrome in horses that results in development of laminitis and is characterized by the presence of general and/or regional adiposity (“cresty neck”), aberrations in blood lipid concentrations, insulin resistance (IR) and/ or hyperinsulinemia. Therapies have focused on improving the state of obesity and insulin resistance with the goal of diminishing the likelihood of laminitis development. A definitive cause for laminitis has not been established, but hyperinsulinemia and IR are likely candidates as experimental states of hyperinsulinemia have been shown to induce laminitis and improvements in insulin sensitivity and obesity have been associated with a decreased risk of laminitis development. This dissertation discusses associations between obesity and IR, as well as potential therapies for alleviating insulin resistance with the ultimate goal of decreasing the risk of developing laminitis. Therapies evaluated included chromium and magnesium, levothyroxine sodium, and metformin hydrochloride. Horses were treated with each supplement for 10 to 36 weeks, depending on the supplement tested, and physical measurements such as body weight, neck circumference, and body condition score were obtained. Throughout each study, blood concentrations of glucose, insulin, and plasma lipids were analyzed. Chromium and magnesium currently do not appear to have any effect on insulin sensitivity, whereas results of levothyroxine administration indicate therapeutic responses, as does metformin, though results indicate further work are required. Research contained in this dissertation focuses on the potential of identifying animals at risk of developing IR and laminitis through measurement of blood biomarkers such as adiponectin and glucagon-like peptide 1. Assays to measure markers included enzyme-linked immunosorbent assays, western blots, and radioimmunoassays. Glucagon-like peptide 1 currently does not appear to differ between healthy and IR animals, but protein band density of high-molecular weight adiponectin does appear to be lower in horses with IR when compared to healthy animals. There is still much to learn about IR in horses, and therapy appears to be dependent on a case by case basis.

Die subklinische Klauenrehe ist eine weltweit vorkommende, multifaktorielle und bei Rindern vor allem nach der Abkalbung gehäuft auftretende Erkrankung. Als prädisponierender Faktor für weitere, z. T. sehr schmerzhafte Klauenerkrankungen wie Sohlengeschwüre oder White-Line-Disease besitzt sie nicht nur eine hohe tierschutzrelevante, sondern auch eine große wirtschaftliche Bedeutung. Die Ätiologie und Pathogenese der Klauenrehe sind bis heute nicht hinreichend geklärt. In neuerer Zeit konzentrieren sich die Forschungen auf den peripartalen Zeitraum und den damit verbundenen metabolischen und hormonellen Einflüssen auf das Klauengewebe. In dieser Arbeit wird der Frage nachgegangen, inwieweit eine erhöhte Fettmobilisation bei Kühen in der negativen Energiebilanz nach der Abkalbung sowie eine verstärkte Lipolyse bei antepartal verfetteten Kühen zur Entstehung der subklinischen Klauenrehe beitragen. Gleichzeitig soll die Frage beantwortet werden, welche Rolle die postpartale Insulinresistenz und die dadurch verminderte Glucoseaufnahme ins periphere Gewebe bei der Ausbildung der subklinischen Klauenrehe spielen. Außerdem soll untersucht werden, inwiefern sich systemische Einflüsse auf die verstärkte Ausbildung der Sohlenhämorrhagien nach der Abkalbung nachweisen lassen und ob diese anhand ausgewählter Blutparameter vorhersehbar sind. Für die Untersuchung wurden 30 primi- und 44 multipare Milchkühe aus drei Betrieben mit Laufstallhaltung ausgewählt. Die Sohlenflächen aller acht Hauptklauen eines Tieres wurden nach funktionellem Klauenschnitt 1 Woche und 8 Wochen p.p. fotografiert, in fünf Zonen eingeteilt und anhand Anzahl, Größe und Schweregrad der sichtbaren Läsionen beurteilt. Zur weiteren Differenzierung wurden verschiedene Klauenscores gebildet. 2 - 3 Wochen vor der Abkalbung sowie 1 Woche, 4 und 8 Wochen p.p. wurden Blutproben entnommen und die Konzentrationen der Freien Fettsäuren (FFS), ß-Hydroxy-Butyrat (BHB), Glucose, Insulin und Haptoglobin bestimmt. Die IGF-1-Konzentration wurde zur weiteren Einschätzung der Energiebilanz bzw. Energieaufnahme und der Körperkondition gemessen. Die Insulinresistenz wurde anhand basaler Insulin- und Glucose-Konzentrationen bestimmt. Des Weiteren wurde vor und nach der Abkalbungdie Rückenfettdicke sonographisch gemessen und die peripartale Fettmobilisierung über die Rückenfettdickenänderung errechnet. Nahezu alle untersuchten Tiere (96 %) zeigten für die subklinische Klauenrehe typische Veränderungen. Bei über der Hälfte der Kühe waren 8 Wochen p.p. an allen vier Gliedmaßen Sohlenhämorrhagien vorhanden. Es konnten signifikante Korrelationen der Klauenscoreparameter sowohl zwischen den einzelnen Zonen als auch zwischen den Hinter- und Vordergliedmaßen gefunden werden. 8 Wochen p.p. war eine signifikante Verschlechterung der Klauengesundheit im Gegensatz zur ersten Woche p.p. zu verzeichnen. Da diese Hämorrhagien erst nach zwei Monaten an der Fußungsfläche sichtbar werden, sind die ursächlichen Veränderungen in der Lederhaut zum Zeitpunkt der Abkalbung entstanden. Dabei waren die lateralen Klauen der Hintergliedmaße an der Rusterholzstelle am stärksten betroffen. Ein Einfluss auf die Klauengesundheit durch die Fütterung, die Bodenbeschaffenheit der Haltungssysteme, altersbedingt sowie durch peripartale Erkrankungen konnte nicht gefunden werden. Ungefähr zwei Drittel der untersuchten Kühe hatten nach der Abkalbung eine negative Energiebilanz. Sie zeigten signifikant weniger Läsionen an den Sohlenflächen als Kühe mit positiver Energiebilanz. Eine übermäßige Rückenfettmobilisierung führte in diesen Untersuchungen nicht zu einer Verschlechterung der Klauengesundheit. Antepartal unterkonditionierte Tiere mit wenig Fettmobilisierung waren sogar stärker von Klauenläsionen betroffen als normal- oder überkonditionierte Kühe. Bei Tieren mit einem peripartalen Abfall der IGF-1-Konzentration waren signifikant mehr Veränderungen an der Sohlenfläche nachzuweisen. Die IGF-1-Konzentration korrelierte dabei aber, im Gegensatz zu vielen Angaben in der Literatur, hochsignifikant negativ mit der Energiebilanz und zeigte keinen Bezug zur RFD, so dass fraglich ist, ob dieser Parameter alleine überhaupt geeignet wäre, eine Aussage über die Energiebilanz oder die Ausbildung einer subklinischen Klauenrehe zu treffen. Bei insulinresistenten Kühen waren eine signifikante Erhöhung der Klauenläsionen sowie ein signifikanter Abfall der IGF-1-Konzentration zu verzeichnen. Die Bestimmung der Insulinresistenz anhand basaler Blutglucose- und Insulin-Konzentrationen bei Kühen nach der Abkalbung erwies sich jedoch als äußerst fragwürdig. Signifikante Korrelationen zwischen den Konzentrationen der einzelnen Blutparameter und den Klauenscoreparametern bestanden, bis auf die FFSKonzentration 2 - 3 Wochen a.p., ausschließlich 1 Woche p.p.. Allerdings blieben die Korrelationen insgesamt relativ niedrig. Die IGF-1-Konzentration korrelierte am häufigsten sowie am engsten, Insulin und Haptoglobin korrelierten zu keinem Zeitpunkt mit den Klauenscoreparametern. Es kann davon ausgegangen werden, dass es sich bei den gefundenen Hämorrhagien an der Sohlenfläche um die Ausbildung einer subklinischen Klauenrehe aufgrund systemischer Einflüsse im peripartalen Zeitraum handelt. Eine negative Energiebilanz sowie die antepartale Verfettung der Milchkühe stellen nach den vorliegenden Untersuchungen keinen Risikofaktor dar. In dieser Arbeit konnte nicht bestätigt werden, dass Lipidmobilisation oder mangelnde Glucoseversorgung nach der Abkalbung eine Rolle bei der Entstehung der subklinischen Klauenrehe spielen.

Made available in DSpace on 2014-06-11T19:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-07Bitstream added on 2014-06-13T19:09:51Z : No. of bitstreams: 1 laskoski_lm_me_jabo.pdf: 647030 bytes, checksum: 950b1f5d5358b98ef4e5d3a67333637b (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A laminite está associada a ativação inflamatória sistêmica, a qual ocorre em diversas afecções, principalmente as gastrointestinais. Sua patogenia este relacionada a ativação de metaloproteinases e degradação do colágeno e/ou hemidesmossomos. 0 objetivo deste estudo foi analisar alterações morfológicas, apoptose, metaloproteinases (MMP) 2 e 9 e infiltração leucocitária, pela pesquisa da lipocalina liberada por neutrófilos (NGAL), no tecido laminar de equinos submetidos a obstrução intestinal experimental. Os animais foram distribuídos nos grupos: controle (Gc), instrumentado (Gi), com procedimento de enterotomia sem obstrução; obstruído tratado com hidrocortisona (Gt) e obstruído não tratado (Gnt). 0 tecido laminar foi analisado pelas técnicas de microscopia em luz, imunohistoquímica e zimografia. Os animals dos grupos experimentais (Gi, Gt e Gnt) demonstraram alterações morfológicas, MMPs e substância formada pela M M P-9 e NGAL superiores aos resultados do Cc. A presença de apoptose não foi evidenciada pelo procedimento cirúrgico. 0 Gc não foi diferente quanto as alterações morfológicas quando comparado ao Gi, e também não diferiu do Gt em relação e MMP-9, principalmente associada a neutrófilos. Com estes resultados, conclui-se que: o modelo experimental pode causar alterações morfológicas associadas a laminite; a apoptose não participa da fase inicial de alterações laminares no córium laminar; a hidrocortisona pode ser utilizada para coibir os efeitos deletérios da ativação inflamatória associada a diversas doenças, sem comprometer o córium laminar. / The laminitis is associated to systemic inflammatory activation that occur in several diseases, mainly gastrointestinal. The pathogeny of the laminitis is related to matrix metalloproteinases (MMPs) activation with collagen or hemidesmossomos degradation. The aim of this study is evaluated of the morphologic alterations, apoptosis, MMPs 2 and 9 and leukocyte infiltration, for research of MMP-9 related to neutrophil gelatinase associated to lipocalin (NGAL) in laminar corium of horses subjected to intestinal obstruction experimental. The animals were divided in groups: control (Cc), instrumented (Gi), with enterotomy without obstruction; obstruction treated by hydrocortisone (Gt) and without treatment (Gnt). The laminar tissue was analyzed by optical microscopy, immunohistochemistry and zymography. The animals showed morphologic alterations, MM Ps expression and MMP-9 associated to NGAL greater the Gc. The presence of apoptosis was not evidend by cirurgical procedure. However, the Gc was not different to Gi about morphologic alterations, and too Gt, about MMP-9, mainly the form conjugated to NGAL. The results indicate that the experimental model can cause morphologic alterations associated with laminitis. The apoptosis is not prerequisite for laminar lesion. The hydrocortisone can be administrated to reduce the deleterious effects of inflammatory activation in several diseases, without to affect the laminar corium.

Submitted by Luanna Matias (lua_matias@yahoo.com.br) on 2015-03-30T11:13:52Z No. of bitstreams: 2 Dissertação - Antônio Dionísio Feitosa Noronha Filho - 2013.pdf: 1195605 bytes, checksum: 6407e61f5c39652f43701aff78788bfd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luanna Matias (lua_matias@yahoo.com.br) on 2015-03-30T12:42:48Z (GMT) No. of bitstreams: 2 Dissertação - Antônio Dionísio Feitosa Noronha Filho - 2013.pdf: 1195605 bytes, checksum: 6407e61f5c39652f43701aff78788bfd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-03-30T12:42:48Z (GMT). No. of bitstreams: 2 Dissertação - Antônio Dionísio Feitosa Noronha Filho - 2013.pdf: 1195605 bytes, checksum: 6407e61f5c39652f43701aff78788bfd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-03-15 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Digital diseases worries livestock industry and arouse interest in extensionists because of the economic losses and difficult control. Losses come from decrease in productive and reproductive performance and, in some cases, the premature culling of high value breeding animals. Besides these, lameness causes negative impact on animal welfare. Digital diseases assume different characteristics in distinct production systems, requiring specific solutions that fits each farms condition. Innovative treatments have been discussed, but are not always economically viable, however, in last years in Veterinary Medicine has increased the interest on the possibility of applying cell therapy in the treatment of diseases related to the locomotor systems, including digital diseases. In the equine, it has been studied its use in the repair of tendineous, articular and osseous lesions, and the perspective of application in other important diseases like laminitis. Considering the importance of digital diseases, especially laminitis, for cattle, cell therapy can represent an additional option for treatment. This study evaluated the development of digital lesions in cattle intensively managed and its response to the treatment with bone marrow mononuclear cells. Were used 15 animals, male, cross-bred (Bos taurus X Bos indicus), healthy, aging about 12 months and with average weight of151,6 ± 24,65 kg, divided in two groups (GT, n=7, GC, n=8). The study was developed in three stages divided in evaluation moments. On first stage, on second month of confinement, after adjustment of management over concrete floor and feeding of corn silage and concentrate, hooves were trimmed and started evaluation of digital disease wich extended to the tenth month. At this time, were observed digital lesions that developed in the period, their influence over behavior and was conducted radiographic examinations trying to establish relationship between external and internal lesions of digits. After ten months of study all animals had its hooves trimmed and all digital lesions, in decorrence of management, were surgically treated. At this time, started last stage in wich animals received cell therapy. To obtain material needed for the protocol, were harvested samples of bone marrow from animals belonging to treatment group (GT). After this, mononuclear cells were isolated and samples were injected at jugular vein. In the protocol for bone marrow harvest and isolation of mononuclear cells was used Ficoll-Paque. On animals from control group (GC), their lesions were treated during hoof trimming, but all anials were evaluated at twelfth month. To certify the effect of cell therapy, besides clinical examination, identification of possible recidivance or new lesions, were done histological examination for occurrence of hyperemia, hemorrhages or edema on dermis, presence of white blood cells inside or outside vessels and necrosis of basal cells from epidermis, and radiographic examination. On direct inspection were observed expressive occurrence of digital lesions like heel horn erosion, haematoma and erosions of sole and white line disease, however with no difference between groups. Major changes in behavior observed along time were decrease in eating time and increase in times lying down and in leisure. Radiographic evaluation didn´t show signs of distal phalanx rotation suggesting external alterations may not be related to internal alterations. Protocols for bone marrow aspiration and for cell isolation using Ficoll-Paque were effective, resulting in obtainance of mononuclear cells with satisfatory viability. But, since it wasn´t observed significative differences between groups related to digital lesions, histological and radiographical examinations, the reoccurrence and appearance of new lesions after cell therapy cannot be related to this treatment, because seem to be more related to management conditions. It can be concluded that applying of bone marrow derived mononuclear cells at jugular vein didn´t show benefits as auxiliary treatment of digital lesions in cattle. The control of risk factors and site of cell application seem to explain the apparent failure of the therapeutic protocol used. / As doenças digitais causam preocupação ao setor produtivo e despertam grande interesse nos técnicos ligados à bovinocultura em função do impacto econômico e do difícil controle. Os prejuízos decorrem de queda no desempenho produtivo, reprodutivo, tratamento oneroso e em alguns casos descarte prematuro de animais de alto mérito genético. Além disso, ameaçam seriamente o bem estar animal. Essas doenças assumem características distintas nos diversos sistemas de produção, o que em alguns casos demandam soluções específicas e tratamentos que atendam a realidade do criatório. Tratamentos inovadores têm sido discutidos, mas nem sempre são viáveis economicamente. Contudo, nos últimos anos, na Medicina Veterinária cresce o interesse na possibilidade de se empregar a terapia celular, que poderia ser aplicada como auxiliar no tratamento de várias enfermidades do aparelho locomotor, incluindo as enfermidades digitais. Em equinos, já se tem investigado seu emprego no reparo de lesões tendíneas, articulares e ósseas, havendo ainda a perspectiva de aplicação em outras doenças importantes como a laminite. Portanto, considerando a importância que as doenças digitais, em especial a laminite, assumem na espécie bovina, a terapia celular pode representar uma ferramenta adicional no tratamento. Esta pesquisa objetivou avaliar o desenvolvimento de lesões digitais em bovinos manejados intensivamente e sua resposta ao tratamento com células mononucleares da medula óssea. Foram empregados 15 bovinos machos, mestiços (Bos taurus X Bos indicus), hígidos com idade inicial aproximada de 12 meses e peso corporal médio de 151,6 ± 24,65 kg, alocados em dois grupos (Grupo tratamento, GT, n=7; Grupo controle, GC, n=8). O estudo foi desenvolvido em três etapas divididos em momentos de avaliação. Na primeira etapa, no 2o mês de confinamento após ajustar o manejo em piso de concreto e a alimentação a base de silagem de milho e concentrado, realizou-se ajuste nas medidas digitais dos animais e iniciou-se o acompanhamento para avaliar a ocorrência de doenças digitais, que se estendeu até o décimo mês. Nesse período, foram realizadas avaliações para diagnosticar as enfermidades que eventualmente surgissem. Estudou-se a influência delas sobre o comportamento diário dos bovinos e procedeu-se a análise macroscópica das lesões digitais e a avaliação radiográfica para se tentar estabelecer uma relação entre as lesões externas e internas dos dígitos. Após dez meses de confinamento, todos os animais tiveram as medidas digitais novamente ajustadas e as enfermidades digitais que surgiram na etapa anterior, em decorrência do manejo, foram tratadas cirurgicamente. Nesse momento iniciou-se a ultima etapa na qual os animais receberam a terapia celular. Para se obter o material necessário que o protocolo exigia colheu-se medula óssea dos bovinos pertencentes ao grupo tratamento (GT), na sequência, as células mononucleares foram isoladas e uma amostra destas foram aplicadas pela veia jugular em cada animal. No protocolo de colheita da medula óssea e isolamento celular empregou-se o Ficoll-Paque®. O grupo controle (GC) foi apenas submetido ao tratamento das lesões e tiveram as medidas dos cascos ajustadas, mas os bovinos de ambos os grupos foram reavaliados ao final do 12o mês do estudo. Para certificar o efeito da terapia celular, além do exame clínico, identificação de possível recrudescimento ou surgimento de novas lesões, realizou-se exame radiográfico, avaliação histológica da região coronária para avaliar presença de hiperemia, hemorragia ou edema da derme, presença de células brancas dentro ou fora dos vasos e necrose de células basais da epiderme. Na avaliação macroscópica foi observada ocorrência expressiva de lesões digitais como erosão de talão, hematoma e erosão de sola e lesão de linha branca, porém, sem diferenças entre grupos. As principais alterações de comportamento observadas ao longo das avaliações foram a redução no tempo de alimentação e aumento nos tempos em ócio e em decúbito, mas, sem diferenças entre grupos tratamento e controle. A avaliação radiográfica não mostrou ter ocorrido a rotação da falange distal indicando que as lesões externas podem não ter relação com as alterações internas. Os resultados obtidos indicam que o protocolo de aspiração de medula óssea e de isolamento celular usando Ficoll-Paque® foi eficaz, resultando na obtenção de células mononucleares com rendimento e viabilidade satisfatórios. Mas, como não foram observadas diferenças significativas entre grupos, tanto com relação as enfermidades como em relação aos resultados histológicos e radiográficos as recidivas ou o aparecimento de novas lesões após a terapia celular não podem ser imputadas a esse tratamento, pois parecem ter relação com o manejo. Conclui-se que a aplicação de células mononucleares da medula óssea pela veia jugular não apresentou benefícios no tratamento auxiliar de lesões digitais em bovinos, sendo que aspectos relacionados ao controle dos fatores de risco e local de aplicação das células parecem explicar a aparente falta de sucesso do protocolo terapêutico empregado.

Einleitung: Lahmheiten bedingt durch Klauenläsionen spielen als Ursache von Tierabgängen in der Schweinehaltung eine sehr große Rolle. Speziell zur Histomorphologie der Laminitis beim Schwein gibt es jedoch kaum Untersuchungen. Ziel der Untersuchungen: Es soll geklärt werden, ob Laminitis bei den Schweinen der untersuchten Population überhaupt auftritt und wenn ja, durch welche Histomorphologie diese Erkrankung gekennzeichnet ist. Gleichzeitig soll untersucht werden, ob sich die Haltung bzw. die Qualität der Muttersau auf Prävalenz und Qualität der Laminitis bei deren Nachkommen auswirken kann. Tiere, Material und Methoden: Untersucht wurden 329 verschieden alte Schweine aus einem Tierversuch mit unterschiedlichen Haltungsbedingungen und Muttersau-Qualitäten. Die Altersgruppen setzten sich wie folgt zusammen: 115 Ferkel am 3. Lebenstag (Saugferkel), 111 Ferkel am 11. Tag nach dem Absetzen (Absatzferkel) sowie 103 adulte Tiere zum Mastende (Mastschweine). Es wurde jeweils die mediale sowie die laterale Klaue einer Gliedmaße untersucht. Unterschieden wurde zwischen Tieren mit normaler und verbesserter Haltung sowie zwischen Tieren mit guter und schlechter Muttersauen-Qualität. Die verbesserte Haltung zeichnete sich durch zusätzliches Heu-/Strohangebot, Mutter-Kind-Beckentränken, Schalentränken sowie durch eine Trinkwasserhygienisierung mittels Chlorbleichlauge aus. Die jeweilige Sauen-Qualitätsgruppe leitete sich von den Ergebnissen der Bonitur der Muttersauen im Hinblick auf Entzündungsanzeichen an den Klauen und der Milchleiste um den 50. Trächtigkeitstag ab und wurde auf die Ferkel übertragen. Das routinemäßig aufgearbeitete Probenmaterial wurde anhand verschiedener Laminitis-typischer Merkmale lichtmikroskopisch untersucht. Zur Auswertung herangezogene statistische Methoden umfassten einen Mann-Whitney-Test (Parameter im Vergleich mit den Haltungs- & Sauen-Qualitätsgruppen) sowie einen Kruskal-Wallis-Test mit „Dunnʾs multiplem post-hoc-Test“ (Parameter im Vergleich mit den drei Altersgruppen). Für den Parameter „Abweichungen von der Lederhaut-blättchenform“ wurden Zusammenhänge zwischen den Haltungs-, Sauen-Qualitäts- und Altersgruppen jeweils mittels einer Häufigkeitstabelle dargestellt. Weiterführend wurden ein exakter Test nach Fisher (Haltung & Sauen-Qualität) sowie ein Chi-Quadrat-Test mit einem exakten Test nach Fisher als post-hoc-Test (Altersgruppen) durchgeführt. Ergebnisse: Die überwiegende Mehrheit der Tiere zeigte Abweichungen von der Lederhautblättchenform im Sinne einer Zergliederung der Lederhautblättchen. Bezogen auf die ausgewerteten Klauen fand sich bei der Mehrzahl eine Mehrfach-Zergliederung. Mastschweine wiesen im Vergleich zu Saugferkeln einen signifikant höheren Anteil an Abweichungen von der Lederhautblättchenform (Zergliederung) auf. Weiterhin nahm die Länge der Lederhautblättchen mit dem Alter signifikant zu. Altersabhängige Effekte zeigten sich auch bei subepithelialen Ödemen, die bei den Saugferkeln im Vergleich zu den anderen Altersgruppen signifikant stärker ausgeprägt waren. Absatzferkel und Mastschweine wiesen hingegen signifikant mehr epitheliale, interzelluläre Ödeme auf. Weiterhin wurden signifikant stärkere Leukozyteninfiltrate bei den Mastschweinen (vornehmlich Lymphozyten) festgestellt. Absatzferkel wiesen im Vergleich zu den Mastschweinen signifikant mehr Thromben auf. Haltungsabhängige Effekte wurden bei der Lederhautblättchen-assoziierten epithelialen Hyperplasie und bei den epithelialen, interzellulären Ödemen beobachtet, wobei jeweils nur die Tiere der verbesserten Haltung signifikante Effekte zeigten. Saugferkel wiesen die höchsten epithelialen Schichtdicken auf. Es zeigten sich signifikant mehr epitheliale, interzelluläre Ödeme bei Absatzferkeln und Mastschweinen. Die Faktoren Alter und Haltung hatten allerdings keinen signifikanten Einfluss auf das Vorkommen von epithelialen, intrazellulären Ödemen und subepithelialen Blutungen. Hinsichtlich der Muttersauen-Qualität zeigten sich keinerlei signifikante Effekte bei den untersuchten Parametern. Weitere untersuchte Parameter wie „Hornkappen“, subepitheliale Fibrose, subepitheliale/epitheliale Nekrose, Hyperämie, intraepitheliale Blutungen, Hämosiderose, Vaskulitis sowie Arteriosklerose wurden nicht nachgewiesen. Mittels PAS-Reaktion fanden sich keine Hinweise auf Basalmembrandegenerationen. Schlussfolgerungen: Laminitis-typische histopathologische Veränderungen waren bei den untersuchten Schweinen nicht nachweisbar. Nichtsdestotrotz wurden wertvolle Erkenntnisse zur Histomorphologie des dermo-epidermalen Übergangs des Wandsegmentes gewonnen, die für weitere Untersuchungen an der Schweineklaue von Nutzen sein werden. Hierbei sind zum einen neu gewonnene Daten zur Schichtdicke des Stratum germinativum in Abhängigkeit vom Alter des Tieres und zum anderen neue Erkenntnisse hinsichtlich der bei einer Mehrzahl der Tiere vorgefundenen Zergliederung der Lederhautblättchen zu nennen.

Das Equine Metabolische Syndrom (EMS) beschreibt einen Cluster von metabolischen Störungen, die durch Adipositas, Insulinresistenz und Hufrehe im Zusammenhang stehen. Ziel dieser Arbeit war es, den Einfluss einer achtwöchigen oralen Supplementierung von Chrom, Metformin beziehungsweise von einer Kombination aus Chrom und Metformin auf die Insulinsensitivität und die Glukosetoleranz bei adipösen Pferden und Ponys zu untersuchen. Für diese Studie standen 24 Pferde und Ponys (14,4 ± 3,87 Jahre, 14 Stuten und 10 Wallache) mit Adipositas, Insulinresistenz und Hufrehe zur Verfügung. Während der achtwöchigen Versuchsphase bekamen die Tiere eine Heuration (1,5 kg Heu/100 kg Körpermasse (KM)) und zweimal täglich das ihnen zugewiesene Supplement (Chrom: 25 µg/kg KM, Metformin: 15 mg/kg KM, Chrom+Metformin: 25 µg/kg KM Chrom + 15 mg/kg KM Metformin in jeweils 25 g Grünmehl/100 kg KM) verabreicht. Eine vierte Gruppe erhielt als Placebo 25 g Grünmehl/100 kg KM ohne Supplement. Vor Versuchsbeginn und nach Versuchsende durchliefen die Pferde und Ponys, an zwei aufeinanderfolgenden Tagen, nach jeweils einer zwölfstündigen Fastenperiode einen kombinierten Glukose-Insulin-Toleranztest (KGIT) zur Bestimmung der Insulinsensitivität und einen oralen Glukose-Toleranz-Test (OGTT) zur Bestimmung der Glukoseabsorption und -toleranz. Im Verlauf des Versuchszeitraums konnte bei den Pferden ein durchschnittlicher Gewichtsverlust von 2,77 ± 2,99 % verzeichnet werden (Behandlung p > 0,05). Beim OGTT zeigte sich keine signifikante Veränderung der Glukose- und Insulinreaktionen zwischen Versuchsbeginn und Versuchsende. Die Seruminsulinmaximalwerte der mit Metformin und der mit Chrom+Metformin supplementierten Gruppen waren nach der Versuchszeit numerisch gesunken (Metformin Versuchsbeginn: 452 ± 642 µU/ml, Versuchsende: 202 ± 121 µU/ml; Chrom+Metformin Versuchsbeginn: 388 ± 347 µU/ml, Versuchsende: 342 ± 164 µU/ml, Behandlung p > 0,05). Im KGIT zeigten sich bei den Glukosewerten keine signifikanten Unterschiede zwischen den einzelnen Behandlungsgruppen und zwischen den Werten vor Versuchsbeginn und nach Versuchsende. Die Seruminsulinkonzentrationen lagen im KGIT vor Versuchsbeginn bei allen Probanden zum Zeitpunkt Minute 45 über 100 µU/ml. Somit gelten alle Versuchsteilnehmer per definitionem als insulinresistent. Nach den acht Wochen Supplementierung befanden sich beim KGIT zum Zeitpunkt Minute 45, mit Ausnahme eines Probanden aus der Chrom+Metformin-Gruppe, alle Seruminsulinwerte weiterhin >100 µU/ml. Somit sind die übrigen 23 Pferde und Ponys weiterhin als insulinresistent einzustufen. Weder Chrom, noch Metformin oder die Kombination von Chrom+Metformin konnte in den hier im Versuch angewandten Dosierungen die Insulinsensitivität und Glukosetoleranz der erkrankten Pferde und Ponys verbessern.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-01-05T14:15:02Z No. of bitstreams: 2 Dissertação - Sabrina Lucas Ribeiro de Freitas - 2015.pdf: 1727922 bytes, checksum: 3502487e7bd9da8dda2d7674ddde726a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-01-06T09:24:49Z (GMT) No. of bitstreams: 2 Dissertação - Sabrina Lucas Ribeiro de Freitas - 2015.pdf: 1727922 bytes, checksum: 3502487e7bd9da8dda2d7674ddde726a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-01-06T09:24:49Z (GMT). No. of bitstreams: 2 Dissertação - Sabrina Lucas Ribeiro de Freitas - 2015.pdf: 1727922 bytes, checksum: 3502487e7bd9da8dda2d7674ddde726a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-09-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The real etiopathogeny of many digital diseases of cattle is still reason to many questions. It can´t be ignored that compromising perfusion and draining of digital vasculature have close relation with some disorders of the digit in this specie. Therefore, a solid knowledge on digital vasculature of the bovine can lead to better understanding of diseases like laminitis. This study intended to decribed venographic findings of the digits of young crossbred (Bos indicus X Bos taurus) cattle, before and after intrarruminal administration of oligofructose as na alternative to the study of laminitis etiopathogenesis a help the study of vascular anatomy of the digits. In the study was used seven crossbred (Bos indicus X Bos taurus) steers, non castrated, aging 12,6±0,9 months and weigthing 156±37,8 kg. Animals were sorted in two groups (GI and GII) receiving 13 and 17 g/kg respectively of intrarruminal oligofructose. Venography were done in the digit of cattle in five different moments, 15 days before (M0), 36 hours (M1), seven (M2), 15 (M3) and 30 (M4) days after intrarruminal administration of high fermentative carboidrate. Venography examination intended to evaluate topographic anatomy and possible vascular changes in the distal pelvic limb of the animals. Description of topographic anatomy was done based on radiographs obtained in all moments and to vascular changes in the distal limb was evaluated diferences among moments, having M0 as control. In the evaluation of vascular changes, main veins and regions like coronary corium were evaluated. Three veins were chosen to measurement of its diameter and data was analysed using ANOVA and T test at 5% significance level. Main veins and regions of coronary region, in diferente moments, were evaluated descriptively. Were identified changes in vascularization at diferente moments, like vasodilation 36 hours after oligofructose administration and vasoconstriction in the other moments, in the main veins and regions of coronary corium and bulbs. Animals from GII, changes were more accentuated. There was statistical difference for common digital dorsal III vein beteween M1 and M4 in each group, with dilation in M1 and constriction in M4. For the palmar branch to the digital cushion of medial digit the existing difference was between the groups in M4, with the constriction of the vein more accentuated in GII. It was concluded that venogtraphic exam of digits in pelvic limbs of young crossbred (Bos indicus X Bos taurus) cattle, examining before and after intrarruminal administration of oligofructose permited description of local vascular anatomy and identified changes compatible with laminits in this specie. / A etiopatogenia de várias doenças podais dos bovinos ainda é motivo de especulação. Mas não se pode ignorar que o comprometimento da irrigação e da drenagem sanguínea dos dígitos tem uma relação estreita com diversas alterações que acometem os dígitos dessa espécie animal. Portanto, o conhecimento sólido da vascularização da extremidade distal dos membros locomotores de bovinos pode nortear o entendimento de enfermidades como a laminite. Esse estudo objetivou descrever achados venográficos dos dígitos dos pés de bovinos jovens, mestiços (Bos indicus X Bos taurus), antes e após aplicação intrarruminal de oligofrutose como alternativas ao estudo da etiopatogenia da laminite e auxiliar no estudo da anatomia vascular dos dígitos. No estudo foram empregados, sete animais mestiços (Bos indicus X Bos taurus), inteiros, com 12,6±0,9 meses de idade e peso de 156±37,8 kg. Distribuiu-se por sorteio os animais em dois grupos (GI e GII) que receberam 13 e 17 g/kg de oligofrutose via intrarruminal, respectivamente. Realizou-se venografia dos dígitos dos bovinos, em cinco momentos distintos, sendo 15 dias antes (M0), 36 horas (M1), sete (M2), quinze (M3) e trinta (M4) dias após aplicação intrarruminal do carboidrato de alta fermentação. A venografia teve a finalidade de avaliar a anatomia topográfica e possíveis alterações vasculares da extremidade distal dos membros pélvicos destes animais. A descrição da anatomia topográfica foi realizada com base nas radiografias obtidas em todos os momentos. Para as alterações vasculares da extremidade distal avaliou-se as diferenças entre os momentos, tendo o M0 como controle. Nesta avaliação veias principais e regiões como a rede do cório coronário foram avaliadas. Escolheu-se três veias para proceder a mensuração dos seus diâmetros e analisá-los utilizando-se a Análise de Variância (ANOVA) e o Teste t, com nível de significância de 5%. As veias principais e da região do cório coronário, nos diferentes momentos, foram avaliados de forma descritiva. Identificaram-se alterações na vascularização nos diferentes momentos, como vasodilatação com 36 horas após aplicação de oligofrutose e vasoconstrição nos demais momentos, nas veias principais e das regiões do cório coronário e rede bulbar. Nos animais do GII as alterações foram mais acentuadas. Houve diferença estatística para a veia digital dorsal comum III entre M1 e M4 dentro de cada grupo, sendo que em M1 estava dilatada e M4 constrita. Para o ramo plantar para o coxim digital do dígito medial a diferença existente foi entre os grupos no momento M4, sendo a constrição desta veia mais acentuada no GII. Concluiu-se que o exame venográfico dos dígitos dos membros pélvicos de bovinos jovens, mestiços (Bos indicus X Bos taurus), realizado antes e após aplicação intrarruminal de oligofrutose possibilitou a descrição da anatomia vascular local e identificou alterações compatíveis com a laminite nessa espécie animal.