No abstract available

Conselho Nacional de Desenvolvimento Científico e Tecnológico / A retrospective study of the goat necropsies performed over a period of 48 years (1964 to 2011) at the Laboratório de Patologia Veterinária (LPV), Universidade Federal de Santa Maria (UFSM), Rio Grande do Sul state, Brazil, was performed. A total of 114 reports of goat necropsies were analyzed. Ninety five necropsies (83.33%) had a conclusive diagnosis and 19 (19.66%) had inconclusive diagnosis. Out of the conclusive cases, infectious and parasitary diseases were the most prevalent, followed, in decreasing order of prevalence, by metabolic and nutritional diseases, poisonings and toxi-infections, and developmental diseases. Hemonchosis was the most prevalent cause of death in this study. Eimeriosis and listeriosis were also important causes of goat deaths. Among the metabolic and nutritional diseases, urolithiasis, osteoporosis, pregnancy toxemia, malnutrition, and white muscle disease were the most prevalent. Mostly the infectious and parasitary diseases and the metabolic and nutritional diseases occurred many times as outbreaks, causing even more important economic losses. Other conditions or lesions that did not fit any of the above groups of diseases affected about 10% of the necropsied goats. In parallel, a study of cases of osteoporosis in goats, diagnosed in the LPV-UFSM, was performed. The epidemiology, clinicopathological changes and possible pathogenetic mechanisms were determined and discussed. Five goats, females, mix breed, with six months to six years of age, that were kept on natural pasture, without supplemental feed, and under overcrowding condition, were affected. The main clinical signs were weight loss, limited mobility, and recumbence for long periods. The main gross bone changes (on the cutting surface) were depletion of cancellous bone (porosity) and marked reduction in the thickness of the cortical bone. Severe serous atrophy of medullary adipose tissue was also observed. Microscopically, in the evaluated regions (proximal humerus, distal radius, distal femur, proximal tibia and lumbar vertebral bodies), moderate to marked reduction in the number and thickness of bone trabeculae in the epiphyses and metaphyses of long bones and in the vertebral bodies were observed. The clinicopathological features indicated that the osteoporosis observed was probably caused by malnutrition. The bone changes (decrease in the number and thickness of trabeculae of cancellous bone) suggest that both mechanisms, poor bone formation and increased bone resorption, contributed to the occurrence of osteoporosis in the goats of this study. / Foi realizado um estudo retrospectivo para determinar a prevalência de doenças de caprinos diagnosticadas no Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM). Para isso, foram examinados os laudos de necropsias de caprinos realizadas num período de 48 anos (1964 a 2011). Foram computados 114 casos de necropsias de caprinos e destes, 95 (83,33%) tinham diagnóstico conclusivo e 19 (19,66%) tinham diagnóstico inconclusivo. Nos casos com diagnósticos conclusivos, as doenças infecciosas e parasitárias foram as mais prevalentes, seguidas em ordem decrescente de prevalência, pelas doenças metabólicas e nutricionais, intoxicações e toxi-infecções e alterações do desenvolvimento. A hemoncose foi a principal causa de morte de caprinos na área de abrangência do LPV-UFSM. Eimeriose e listeriose também foram causas importantes de morte. Dentre as doenças metabólicas e nutricionais, urolitíase, osteoporose, toxemia da prenhez, desnutrição e doença dos músculos brancos foram as mais prevalentes. Principalmente as doenças infecciosas e parasitárias e as metabólicas e nutricionais ocorreram muitas vezes na forma de surtos, acarretando maiores perdas econômicas associadas. Outras alterações de diferentes naturezas e etiologias que não se enquadravam nos grupos de doenças acima afetaram cerca de 10% dos caprinos examinados. Paralelamente foi realizado um estudo de casos de osteoporose em caprinos, diagnosticados no LPV-UFSM, no qual foram determinados a epidemiologia, o quadro clínico-patológico e os prováveis mecanismos patogenéticos envolvidos. Cinco cabras, fêmeas, SRD, de seis meses a seis anos de idade, que eram mantidas em campo nativo, sem suplementação com ração e com superlotação foram afetadas. Os principais sinais clínicos foram emagrecimento progressivo, dificuldade de locomoção e permanência em decúbito por longos períodos. As principais alterações ósseas macroscópicas nos ossos examinados (superfície de corte) caracterizavam-se por depleção do osso esponjoso (porosidade) e redução acentuada da espessura do osso cortical. Havia também marcada atrofia serosa da gordura da medula óssea. Microscopicamente, nas regiões avaliadas (úmero proximal, rádio distal, fêmur distal, tíbia proximal e corpos das vértebras lombares) foi observada redução moderada a acentuada do número e espessura das trabéculas ósseas nas epífises e metáfises dos ossos longos e nos corpos vertebrais. Os achados clínico-patológicos indicaram que a osteoporose observada provavelmente foi causada por desnutrição. As alterações ósseas (diminuição no número e na espessura das trabéculas do osso esponjoso) sugerem que ambos os mecanismos, má formação óssea e reabsorção óssea aumentada, contribuíram para a ocorrência de osteoporose nos caprinos deste estudo.

M.S.

Rhinoceroses were an important part of North America’s Paleogene and Neogene ecosystems, with Teleoceras aepysoma being one of the last representatives of this family. Specimens of T. aepysoma from the Gray Fossil Site (GFS) possess distinct/peculiar pathologies: including a pair of fused ribs and ankylosed phalanges. A qualitative description of the pathologies in the GFS T. aepysoma, including new material, was conducted to accurately identify pathologies and make interpretations about the life history of the GFS rhinos. Analysis suggests that rheumatoid arthritis is common in the lower limb bones of GFS rhinos. Additionally, the rib and toe pathologies are more severe than anticipated, with the ribs showing multiple stages of healing indicating repeated trauma, likely being the first direct evidence of agonistic behavior in Teleoceras. This study provides a glimpse of what pathological conditions rhinocerotids may have been vulnerable to through time, as well as a baseline for future studies.

Degenerative joint disease, or osteoarthritis, is a major cause of lameness and morbidity in horses, humans, and dogs. There are no truly satisfactory cures for this widespread problem and current treatments all have limitations or unwanted side effects. New cell-based strategies to repair joint surface lesions have generated a high level of interest, but have yet to achieve the full restoration of articular cartilage structure and function. Currently used therapy cells include autologous chondrocytes and adult mesenchymal cells such as bone marrow derived cells and adipose derived cells. Unfortunately, the resultant repair tissue is biomechanically inferior fibrocartilage. A critical gap in knowledge in this regard is a limited understanding of the specific cellular phenotype of normal, robust articular chondrocytes. This thesis examines the global mRNA transcriptome of equine articular cartilage to test the hypothesis that adult articular chondrocytes have a unique gene expression profile. In the first part of the study, RNA-sequencing was used to compare the mRNA transcriptome of normal adult articular cartilage with five other cartilaginous tissues. From these comparisons, locus level gene expression and alternative splicing patterns have been identified that clearly distinguish articular cartilage. In the second part of the study, fetal (interzone, cartilage anlagen chondrocytes, dermal fibroblasts) and adult (bone marrow derived, adipose derived, articular chondrocytes, dermal fibroblasts) primary cells were grown in culture and stimulated to differentiate into chondrocytes. The chondrogenic differentiation potential as assessed by matrix proteoglycan and the expression of cartilage biomarker genes was highly variable among cell types. Together, these results advance our understanding of the specific phenotype of articular chondrocytes and the potential of prospective therapeutic progenitor cells to differentiate into articular chondrocytes. This new knowledge will improve efforts to optimize cell-based therapies for osteoarthritis and the repair of joint cartilage lesions.

Herpes simplex virus (HSV) infection of the cornea leads to a blinding immuno-inflammatory condition of the eye also called stromal keratitis (SK). SK immunopathology is characterized by the infiltration of CD4+ T cells of Th1 phenotype as well as the development of new blood vessels into the normally avascular cornea. Studies in mouse models of SK have firmly established the role of CD4+ T cells, and particularly of Th1 phenotype, as the principal mediators of SK immunopathology. However, with the recent discovery of IL-17A and Th17 cells, the role of this cytokine as well as Th17 cells remains to be further defined. Recently it was shown that the normal cornea expresses VEGF-A, however its biological activity is impeded by its binding to a soluble form of VEGF-A receptor-1 (sVEGFR-1). Past studies have implicated the role of vascular endothelial growth factor-A (VEGF-A) in HSV induced corneal angiogenesis, however the source of VEGF-A as well as molecular mechanisms, particularly in the context of VEGF-A/sVEGFR-1 balance during HSV infection, are poorly understood. The first part of this dissertation (I) reviews past literature on HSV induced corneal SK immunopathology. It focuses on the understanding of HSV-1 induced events that particularly results in corneal angiogenesis as well as tissue damage mediated by different type of cells as well as their secreted products. The next three parts (II-IV) focus on the mechanisms of HSV induced corneal angiogenesis as well as the relative role of Th1 and Th17 cells in SK immunopathology. Results in part II focuses on the relative role of IFN-γ/IL-17 as well as Th1/Th17 cells in HSV induced corneal immunopathology. The third section evaluate the significance of VEGF-A/sVEGFR-1 balance in HSV induced corneal neovascularization. Results in part IV focus on the role of IL-17A in altering the balance between VEGF-A and sVEGFR-1 post ocular HSV infection and subsequent corneal angiogenesis. Collectively these studies identified novel mechanisms by which HSV infection of the cornea leads to the development of angiogenesis as well as corneal tissue damage and subsequent SK immunopathology, the most common cause of infectious blindness in the Western World.

A naturally occurring disease involving hyperplasia of the thyroid gland and a consistent pattern of musculoskeletal deformities of newborn foals in western Canada was first described in 1981. This disease was an important cause of foal mortality and, therefore, reproductive loss throughout western Canada during the 1990s and has since been recognized in western Ontario and the northwestern United States. A series of investigations were conducted to describe, characterize, and attempt to determine the pathogenesis and cause of this syndrome. Affected foals were typically born after a long gestation (x = 360 days, range = 340 to 400 days), were diagnosed as hypothyroid based on a poor response to the administration of thyroid-stimulating hormone, and had various musculoskeletal lesions of which mandibular prognathism, flexural deformities and rupture of tendons of the limbs, and incomplete ossification of the carpal and tarsal bones were present most commonly. In spite of the normal to long gestation, foals had signs of immaturity, were usually weak and unable to stand, became septic, and died or were euthanatised. Similar histories, clinical findings, and lesions were present in surgically created hypothyroid foals that were thyroidectomized in utero at about 210 days gestation. These findings supported the conclusion that foals which naturally developed these lesions were also hypothyroid in utero and that all the lesions present in affected foals were the result of the hypothyroidism and not of an underlying concurrent disease process. A case-control study was conducted to identify risk factors for naturally occurring congenital hypothyroidism. Information from congenitally hypothyroid foals concerning foal and dam signalment, farm environment, and dam management was compared with that from normal foals. Pregnant mares fed greenfeed, not supplemented with mineral, that left their "home farm" during gestation, or grazed irrigated pasture, had a 13.1 (<i>P</i>=0.0068), 5.6 (<i>P</i>=0.0472), 4.3 (<i>P</i>=0.0076) and approximately 15.3 (<i>P</i>=0.0245) times greater odds, respectively, of producing a congenitally hypothyroid foal than mares not exposed to these factors. Greenfeed often contains high levels of nitrate (NO₃^-) which is known to impair thyroid gland function. In light of this, forage samples from participating farms were analysed for nitrate levels. The odds of one or more congenitally hypothyroid foal being born on a farm feeding forage with at least a trace of nitrate was 8.0 times greater (<i>P</i>=0.0873) than the odds of the disease occurring a farm that fed forage free of nitrate. Further, the odds of a mare producing an affected foal when fed forage containing at least a trace of nitrate was 5.9 times greater (<i>P</i>=0.0007) than a mare fed nitrate-free forage.This study suggests that congenital hypothyroidism in foals may result from diets containing nitrate or low in iodine being fed to pregnant mares. These results need to be confirmed through further field investigations and controlled experiments. However, if they are accurate, there is cause for concern that other livestock raised in areas where congenitally hypothyroid foals occur may be exposed to the same dietary risk factors and may suffer similar disease.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This thesis was composed of three scientific papers. The first paper was the more comprehensive. All lesions of the urinary system of dogs necropsied between 1999 and 2010 were analyzed. In this time frame, 3,189 dogs were necropsied at the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) and 1,063 had lesions in the urinary system. In most of the dogs (79.1%), lesions were single and in about 21% they were multiple, totalizing 1,373 lesions. Kidneys were 2.8 times more affected than the lower urinary tract (LUT). One third of the lesions in the urinary system were cause of spontaneous death or reason for euthanasia (SD/EUTH) of the affected dogs. The other lesions were considered incidental findings. The main renal lesions diagnosed were: tubulointerstitial nephritis, infarct, granulomatous nephritis, glomerulonephritis, metastatic/multicentric neoplasms, pyelonephritis/pyelitis, and hydronephrosis. The main LUT lesions were: cystitis, presence of viral inclusions bodies, urolithiasis, urinary bladder dilatation, urinary bladder rupture, and metastatic/multicentric neoplasms. Epidemiological aspects such as gender, breed, and age of affected dogs had significant variations according to the type of lesion diagnosed. Uremia was observed in a significant number of cases of SD/EUTH and was mostly due to renal lesions. The second paper was focused on urolithiasis. From 1990 to 2010, 4,872 dogs were necropsied at LPV-UFSM. Out of these, 1.5% had uroliths along the urinary tract. The epidemiological profile of the affected dogs showed predominance of males; adults; and pure breeds. Clinical signs suggestive of urolithiasis were informed in 30.3% of the dogs and consisted mainly of hematuria, anuria, dysuria and urinary incontinency. The uroliths were found in one or more anatomical sites, and the main affected ones, in descending order, were urinary bladder, kidney, and urethra. Secondary lesions to urolithiasis were observed in about 40% of the cases. The most prevalent were cystitis, urethral obstruction, hydroureter, hydronephrosis, urinary bladder rupture, and pyelonephritis. In 25% of the affected dogs, SD/EUTH occurred due to the secondary lesions of urolithiasis. Extra-renal lesions of uremia were observed in 9 out of the 76 cases. The third article was focused on urinary system neoplasms of dogs. The main purposes of this study were to establish the prevalence and types of primary and metastatic/multicentric neoplasms of the urinary system in dogs submitted to necropsy in about 21 years at the LPV-UFSM. Neoplasms of the urinary system were present in 113 (2.4%) dogs. Twenty seven were primary neoplasms and 86 were metastatic or part of a multicentric tumor. The majority of the primary neoplasms were of epithelial origin. Thirteen dogs had primary renal neoplasms (prevalence of 0.27% over all dogs necropsied). Cystadenocarcinoma/cystadenoma and renal cell carcinoma were the most prevalent primary renal neoplasms and transitional cell carcinoma was the most prevalent urinary bladder neoplasm. Metastatic or multicentric tumors affecting the urinary system were the most prevalent (76.1%); mesenchymal tumors were more common than epithelial tumors; most of them were localized in the kidney. Metastases of mammary tumors and multicentric lymphoma were the most prevalent histologic types. / Esta tese foi constituída de três partes que resultaram em três artigos científicos. O primeiro artigo foi o mais abrangente, onde todas as lesões do sistema urinário em cães necropsiados entre 1999 e 2010 foram analisadas. No período estudado foram necropsiados 3.189 cães no Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) e destes, um terço (1.063) apresentou lesões no sistema urinário. Na maioria (79,1%) dos cães foram observadas lesões únicas e em aproximadamente 21% havia lesões múltiplas, totalizando 1.373 lesões. Os rins foram 2,8 vezes mais afetados que o trato urinário inferior (TUI). Um terço das lesões no sistema urinário foram causa de morte espontânea/eutanásia (ME/EUT). As demais foram consideradas achados incidentais. As principais lesões renais diagnosticadas foram: nefrite túbulo-intersticial, infarto, nefrite granulomatosa, glomerulonefrite, neoplasmas metastáticos/multicêntricos, pielonefrite/pielite e hidronefrose. As principais lesões do TUI diagnosticadas foram: cistite, presença de inclusões virais, urolitíase, dilatação da bexiga, ruptura de bexiga e neoplasmas metastáticos/multicêntricos. As características epidemiológicas como sexo, raça e idade dos cães afetados tiveram variações significativas de acordo com o tipo de lesão diagnosticada. Uremia foi observada em um número significativo de casos de ME/EUT e foi, principalmente, secundária a lesões renais. O segundo artigo enfocou a urolitíase. No período analisado (1990 2010) foram necropsiados 4.872 cães. Destes, 1,5% apresentaram urólitos ao longo do sistema urinário. O perfil epidemiológico mostrou o predomínio de cães machos; adultos; e com raça definida. Sinais clínicos indicativos de urolitíase foram computados em cerca de um terço dos casos e consistiram principalmente de hematúria, anúria, disúria e incontinência urinária. Os urólitos tiveram localização única ou múltipla e os locais anatômicos mais frequentemente acometimentos, em ordem decrescente de prevalência, foram bexiga, rim e uretra. Lesões secundárias à urolitíase foram observadas em aproximadamente 40% dos cães afetados. As mais prevalentes foram cistite, obstrução uretral, hidroureter, hidronefrose, ruptura vesical e pielonefrite. Em 25% do total de cães afetados ocorreu ME/EUT decorrentes das lesões secundárias à urolitíase. Uremia foi observada em 9 dos 76 casos. O terceiro artigo enfocou os neoplasmas que acometem o sistema urinário de cães. Nesse estudo foram determinados a prevalência e os tipos de neoplasmas primários e metastáticos/multicêntricos que ocorreram no sistema urinário de cães recebidos para necropsia no LPV-UFSM num período de aproximadamente 21 anos (janeiro de 1990 julho de 2010). Em 113 (2,4%) dos cães necropsiados no período estudado, foram diagnosticados 27 neoplasmas primários e 86 metastáticos ou como parte de tumores multicêntricos no sistema urinário. Dos neoplasmas primários, a grande maioria teve origem epitelial. Treze casos eram neoplasmas renais primários (0,27% do total de cães necropsiados). Cistadenocarcinoma/cistadenoma e o carcinoma de células renais foram os neoplasmas primários mais prevalentes no rim e o carcinoma de células de transição foi o mais prevalente na bexiga. Os neoplasmas metastáticos e multicêntricos que afetaram o sistema urinário foram os mais prevalentes (76,1%), com predomínio mesenquimal. Destes, a grande maioria estava localizada no rim e, quanto ao tipo histológico, as metástases de neoplasmas mamários e o linfoma multicêntrico predominaram.

<p>Despite previously accepted dogma, several recently published studies in humans and mice have shown that mature red blood cells (RBCs) contain a pool of microRNAs. Their role is currently uncertain; however, it has been suggested that microRNAs may play a role in cellular communications as they can be transferred from the RBC to endothelial cells or other cells. This thesis investigated the set of differentially abundant microRNAs found in mature RBCs of felines with oxidative stress, using diabetes mellitus as an oxidant stressor. We postulated that individual microRNAs identified in this study, might be valuable targets for future studies, investigating the role specific microRNAs play in the development or progression of diabetes and in the oxidative damage inflicted on the red cell and other cells by this disease.</p><p>The first specific objective of this thesis was to document oxidative stress in diabetic cats. In the absence of validated assays to document the presence of oxidative protein damage in felines, we first evaluated the performance of a commonly used colorimetric assay for measurement of protein carbonyls (PC) in serum and plasma. Although within run variation was acceptable and performed well over a wide range of PC content values, there were severe limitations related to excessive between run-variation, hemoglobin interference, and difficulty of assay performance. Therefore, we developed and validated a new method, using a fluorescent probe. This new assay had good within and between-run variations, a broad analytical range, and was easy and rapid to run. Hemoglobin and triglyceride only affected the results when present at moderate to higher levels. To further evaluate their redox status, free-radical production and oxidative stress were measured in diabetic and healthy, control cats. The presence of oxidative stress was assessed by measurement of the resulting damage to biomolecules, and detection of antioxidant levels. Our data indicated the presence of protein and membrane lipid oxidation in diabetic individuals and suggest that the redox status of the mature RBC was shifted toward an oxidation state.</p><p>In the final chapter of this thesis, we document the presence of an abundant and diverse set of microRNAs in the mature erythrocytes of healthy and diabetic cats. While their function in the mature erythrocyte remains unknown, a difference was found in the microRNA expression patterns of diabetic and healthy cats. Our data uncovered severe bias in the microRNA sequencing such that the expression levels of some microRNAs appeared to be artificially increased and other diminished. The library construction kit used, appeared to be the cause of this bias. Among the 899 erythrocyte microRNAs sequenced, 12 differentially abundant microRNAs were identified in diabetic cats, however only 6 were differentially abundant by RT-qPCR. Let-7b, miR-1692, miR-339, miR-486 and a feline specific microRNA were increased in mature RBCs of diabetic cats, while miR-451 was decreased.</p><p><a></a><a></a>In conclusion, we have shown that diabetic cats have evidence of significant systemic protein and lipid oxidation as well as erythrocytic oxidative stress. The new, fluorescent PC content assay developed and validated herein could serve as useful tool to better understand the role and consequence of oxidative stress in feline diabetes or other diseases and to monitor antioxidant treatment. Further, this test could be readily adapted for use in other domestic species. Additionally, we have shown that a set of erythrocytic microRNAs are differently abundant in diabetic in comparison to healthy cats. The significance of such changes is currently uncertain. It could represent adaptation of erythroid precursors to changes in their environment during erythropoiesis and as such, these microRNAs may be useful biomarkers for altered hematopoiesis. If microRNAs play a role in communication between circulating mature RBCs and cells in their surroundings such as endothelial cells, the possibility that changes in their expression in this host cells may result in pathology is an intriguing possibility that need to be further explored.</p>

<p dir="ltr">Viruses have long been investigated for their possible associations with a multitude of neurodegenerative diseases. Recently, herpesviruses, such as human herpesvirus 6 (HHV-6) and Epstein-Barr virus (EBV), have been of growing interest as potential triggers or contributors in the pathogenesis of Alzheimer’s disease (AD) and other causes of dementia. The role that herpesviruses play in these disease processes remains unclear, with evidence for the infectious hypothesis of AD often ambiguous and strongly debated. Obtaining definitive evidence for the role of herpesviruses in AD and other dementia-related diseases remains challenging due to many factors, including the complexities of multifactorial diseases, the unique lifecycle of herpesviruses, and the lack of translational <i>in vivo</i> models of herpesvirus infection and AD. We investigated the potential association of herpesviruses with these diseases through the application of bioinformatic analyses and molecular techniques to human whole genome sequences (WGS) and the utilization of a nonhuman primate model, the common marmoset <i>(Callithrix jacchus).</i></p><p dir="ltr">We observed a higher prevalence of HHV-6 partial integration in synucleinopathies associated with dementia compared to other cohorts. We characterized Callitrichine herpesvirus 3 (CalHV-3) in the common marmoset as a translational model of gammaherpesvirus infection, highlighting similarities to human EBV infection. We described beta-amyloid (Ab) pathology in the brains of herpesvirus infected and uninfected marmosets. In marmosets infected with CalHV-3, we noted lower plasma Ab42:40 ratios and GFAP levels and a positive correlation between viral load and Ab42, Ab40, and total tau. These findings suggest that herpesviruses may play a role in dementia-related neurodegenerative diseases through multiple different mechanisms, including the integration of viral genomes and the alteration or augmentation of disease biomarkers.</p>