Conselho Nacional de Desenvolvimento Científico e Tecnológico / Seven outbreaks and an isolated case of meningoencephalitis caused by bovine herpesvirus-5 (BoHV-5) in cattle from Rio Grande do Sul, Brazil, occurring in 2002-2004 are described. From a total population at risk of 1,359 cattle, 54 1-18-month-old calves from both sexes and several breeds were affected and 50 died spontaneously or were euthanatized while moribund. The highest frequency of cases were in recently weaned calves or calves submitted to other stressing factors. General rates of morbidity, mortality and lethality were respectively 3.97%, 3.67% e 92.59%. Clinical courses varied from 3-10 days and included depression, nasal and ocular discharge, grinding of teeth, circling, blindness, fever, nistagmus, trembling, anorexia, dysphagia, drooling, incoordination, head pressing, rough hair coat, tachycardia, tachypnea, abdominal pain, melena, falls, recumbency, opisthotonus, convulsions and paddling. Nineteen calves were necropsied. Necropsy findings were characterized by hyperemia of leptomeninges, swollen of rostral portions of the telencephalon, and flattening of frontal lobes gyri; frequently in these frontal areas there were segmental brown-yellow discoloration and softening (malacia) of the cortex. In cases with more protracted clinical courses there were extensive swelling, softening and hemorrhaging of the telencephalic frontal lobes. Microscopically, all affected cattle had a necrotizing non-suppurative meningoencephalitis with variable distribution among the 19 cases and among the various telencephalic regions of the same case. The severity of these changes were more marked, in decreasing order of intensity, in the telencephalic frontal cortex, basal ganglia (nuclei), thalamus, brain stem, parietal telencephalic cortex, occipital telencephalic cortex and cerebellum. Perivascular inflammatory infiltrate consisted predominantly of lymphocytes, plasm cells, and, less frequently, neutrophils. Additional microscopic findings included variable degrees of gliosis, edema, neuronal necrosis in the telencephalic cortex characterized by shrinking and eosinophilia of perikaria and nuclear picnosis (red neuron); basophilic intranuclear inclusion bodies in astrocytes and neurons (21.05% of the cases); sattelitosis; and neuronophagia. The areas of softening in the cortical substance consisted of necrosis of the neuroctodermal elements with maintenance of mesenchymal structures (vessels and microglia), infiltrate of gitter cells, and, in more severe cases, extensive hemorrhages. In chronic cases, only vascular structures and a few gitter cells remained in the cortical area leaving a cavity between white matter and leptomeninges (residual lesion). The diagnosis was based on the epidemiological, clinical, necropsy and histopathologial findings. Viral isolation, immunofluorescent antibody technique and polymerase chain reaction were performed in three outbreaks. / São descritos sete surtos e um caso isolado de meningoencefalite por herpesvírus bovino-5 (BoHV-5) em bovinos no Rio Grande do Sul entre 2002-2004. Foram afetados bovinos de 1-18 meses, de diversas raças e ambos os sexos. A maior freqüência foi observada em bovinos recém-desmamados e submetidos a outros fatores de estresse. Nesses surtos, de uma população total sob risco de 1.359 bovinos, 54 foram afetados, quatro se recuperaram e 50 morreram espontaneamente ou foram submetidos à eutanásia quando moribundos. Os índices gerais de morbidade, mortalidade e letalidade foram, respectivamente, de 3,97%, 3,67% e 92,59%. A evolução clínica variou de 3-10 dias e os sinais eram caracterizados por depressão, corrimento nasal ou ocular, ranger de dentes, andar em círculos, cegueira, febre, nistagmo, tremores, anorexia, disfagia, sialorréia, incoordenação, pressão da cabeça contra objetos, pêlos arrepiados, taquicardia, taquipnéia, dor abdominal, melena, quedas, decúbito, opistótono, convulsões e movimentos de pedalagem. Dezenove bezerros foram necropsiados. Achados de necropsia foram caracterizados por hiperemia das leptomeninges, tumefação das porções rostrais do telencéfalo, com achatamento das circunvoluções dos lobos frontais; nessas áreas havia focos marromamarelados e amolecidos (malacia). Nos casos de evolução clínica mais longa era observada acentuada tumefação, amolecimento e extensas áreas de hemorragia nos lobos frontais telencefálicos. Microscopicamente, todos os bovinos afetados apresentaram meningoencefalite não-supurativa e necrosante, que variou quanto à localização e intensidade nos 19 casos examinados e nas seções de encéfalo de um mesmo caso. A intensidade dessas lesões foi mais acentuada, em ordem decrescente, no córtex telencefálico frontal, nos núcleos da base, tálamo, tronco encefálico, córtex parietal, córtex occipital e cerebelo. O infiltrado inflamatório perivascular era constituído predominantemente por linfócitos e plasmócitos e, menos freqüentemente, neutrófilos. Outros achados incluíam variados graus de gliose, edema, necrose neuronal no córtex telencefálico, caracterizada por encarquilhamento e eosinofilia do citoplasma e picnose nuclear (neurônio vermelho), corpúsculos de inclusão basofílicos intranucleares em astrócitos e neurônios (21,05% dos casos), satelitose e neuronofagia. As áreas de amolecimento do parênquima eram caracterizadas por necrose do componente neuroectodérmico e manutenção das estruturas mesenquimais (vasos e micróglia), com infiltrado de células gitter e, em casos mais graves, áreas de hemorragia. Nos casos crônicos apenas estruturas vasculares e poucas células gitter permaneciam, formando uma cavidade entre a substância branca e as leptomeninges (lesão residual). O diagnóstico foi realizado com base nos achados epidemiológicos, clínicos, de necropsia e histopatológicos. Adicionalmente, foi realizado isolamento viral, imunofluorescência e reação em cadeia da polimerase em três surtos.

Cases of trypanosomiasis by Trypanosoma evansi were diagnosed in horses in the state of Rio Grande do Sul, Brazil, between 2003 and 2006. In one stud farm (Farm A) with 125 horses, 53 died. Additionally, around 80 mares were sent to Farm A to be bred. Of those, 66 became ill and 56 died after being returned to their farms of origin. Twenty three horses clinically affected by the disease were observed. Clinical signs included loss of weight (despite voracious appetite), lethargy, incoordination and instability of hindlimbs, atrophy of the large muscles of the hindlimbs, muscle weakness and paleness of mucosae. Specimens of T. evansi were detected in the blood drawn from four affected horses. Normocytic normochromic anemia with PCVs ranging from 15 to 31%, leucocytosis due to lymphocytosis associated to large atypical lymphocytes was observed in several affected horses. High levels of antibodies against T. evansi were detected in the serum of fifteen horses. Ten horses presented encephalic neurological signs such as circling, ataxia, blindness, excitation, falls, listlessness, proprioception deficits and head tilt. One horse assumed a dog-seating position . Necropsy findings included muscle atrophy, enlargement and lymphoid hyperplasia of the spleen and lymphnodes. Seven out of the 9 necropsied horses with encephalic signs had asymmetrical gross lesions in the brain consisting of flattening of gyri and focal extensive areas of yellow discoloration and softening of white matter. Histologically, an overwhelming necrotizing anencephalitis was observed in all 9 horses with encephalic neurological signs. This panencephalitis was characterized by marked edema, demyelination and malacia, and perivascular infiltrates of up to 20 rows of mononuclear cells affecting mainly the white matter. Several plasma cells in the inflammatory infiltrate contained numerous eosinophilic globules (Mott cells) or homogenous bright-red material (flame cells) in their cytoplasm. Mild to moderate meningomyelitis and/or meningitis were observed in the spinal cord of 5 horses. Similar histological lesions were observed in the spinal cord of the horse with the dog-seating position . The brains of nine horses with the encephalic signs were submitted to immunohistochemistry stain by the streptavidin-biotin technique. In eight brains moderate to abundant specimens of T. evansi in the perivascular spaces and neuropile were marked by the specific antibody. / Casos de tripanossomíase por Trypanosoma evansi foram diagnosticados em eqüinos no Rio Grande do Sul entre 2003 e 2006. Em uma propriedade (Propriedade A) com 125 eqüinos, 53 morreram. A Propriedade A recebeu ao redor de 80 éguas de outras propriedades para cobertura. Dessas, 66 adoeceram e 56 morreram após voltarem para suas propriedades de origem. A doença clínica observada em 23 eqüinos caracterizava-se por emagrecimento (apesar de apetite voraz), letargia, incoordenação e instabilidade dos membros pélvicos, atrofia das grandes massas musculares dos membros pélvicos, fraqueza muscular e palidez das mucosas. Exemplares de T. evansi foram observados na corrente sangüínea de 4 eqüinos. Anemia normocítica normocrômica, com hematócritos que variavam de 15-31%, e leucocitose por linfocitose associada à presença de linfócitos atípicos foram observadas em vários eqüinos. Altos níveis de anticorpos contra T. evansi foram detectados em 15 eqüinos. Dez eqüinos desenvolveram um quadro neurológico encefálico caracterizado por andar em círculos, ataxia, cegueira, hiperexcitabilidade, quedas, embotamento, déficits proprioceptivos e desvio da cabeça. Um eqüino desenvolveu posição de cão sentado . Nas 15 necropsias, havia esplenomegalia, linfadenomegalia, hiperplasia linfóide no baço e linfonodo e atrofia das grandes massas musculares dos membros pélvicos. Sete dos nove eqüinos com um quadro neurológico encefálico que foram necropsiados apresentavam lesões encefálicas macroscópicas assimétricas que consistiam de achatamentos dos giros e áreas amarelas e amolecidas focalmente extensas na substância branca. Histologicamente, uma panencefalite necrosante avassaladora foi observada em todos os 9 eqüinos. Essa panencefalite era caracterizada por acentuado edema, desmielinização, malacia e infiltrado perivascular de até 20 fileiras de células mononucleares afetando principalmente a substância branca. Vários plasmócitos no infiltrado inflamatório continham numerosos glóbulos eosinofílicos (células de Mott) ou material vermelho-brilhante (células em flama) em seus citoplasmas. Meningomielite e/ou meningite leve ou moderada foram observadas na medula espinhal de 5 eqüinos. Lesões semelhantes foram observadas na medula espinhal do eqüino que desenvolveu posição de cão sentado . Os encéfalos de 9 eqüinos com quadro encefálico foram submetidos à técnica de imunoistoquímica estreptoavidina-biotina; em oito observou-se a marcação de números moderados ou elevados de espécimes de T. evansi pelo anticorpo específico nos espaços perivasculares e na neurópila.

Many neurodegenerative diseases are characterized by the accumulation of misfolded proteins that often share common morphological and biochemical features, and can similarly co-localize with several other proteins, including various chaperone proteins. Chaperone proteins, like heat shock protein 27 (HSP27), heme oxygenase 1 (HO-1) and clusterin, have been implicated as potent modulators of misfolded proteins, thus may play important roles in the pathogenesis of neurodegenerative diseases. The present study aims to investigate their roles in the pathogenesis of Frontotemporal lobar degeneration (FTLD), Alzheimer's disease (AD), Parkinson's disease (PD), and Motor neuron disease (MND) by determining their distribution and amount via immunohistochemical staining and western blotting in diseased and control subjects.There were distinct patterns of HSP27 and clusterin immunostaining in different brain regions. For HSP27, patients with AD and FTLD were in general more severely affected than were patients with MND and control subjects. For clusterin, patients with AD and FTLD were more severely affected than control subjects where neurons and glial cells were concerned, while patients with AD and control subjects were more severely affected than those with FTLD where diffuse and cored plaques were concerned. However, there were no obvious differences in the pattern of HO-1 immunostaining in various brain regions in patients with AD or FTLD relative to control subjects. Moreover, there was no association between HSP27, HO-1 and clusterin with disease or histological type, and the ‘classic’ neuropathological changes in FTLD, AD and MND were not immunoreactive to any of these proteins. There were significant correlations between the degrees of HO-1 and clusterin immunostaining in many brain areas for both AD and FTLD cases, and for all cases overall, but none between HSP27 and clusterin or HSP27 and HO-1. Present results suggest an involvement with ongoing cellular stress, misfolded or unfolded protein accumulation or the deficits/failure of other relevant protein quality control systems, in the pathogenesis of these neurodegenerative diseases. Present work may therefore have implications for the further development of ideas concerning the cause or treatment of neurodegenerative diseases where there is aberrant accumulation of misfolded, aggregated protein, and perhaps for conformational diseases in general. However, there are still many issues remain to be elucidated. Further research aimed at understanding the function and mechanisms of the chaperone system, and other protein quality control mechanisms, in the pathogenesis of neurodegenerative diseases is still needed.

Anxiety and depression are highly prevalent diseases with common heritable elements, but the particular genetic mechanisms and biological pathways underlying them are poorly understood. Part of the challenge in understanding the genetic basis of these disorders is that they are polygenic and often context-dependent. In my thesis, I apply a series of modern statistical tools to ascertain some of the myriad genetic and environmental factors that underlie fear-related behaviours in nearly two thousand heterogeneous stock mice, which serve as animal models of anxiety and depression. Using a Bayesian method called Sparse Partitioning and a frequentist method called Bagphenotype, I identify gene-by-sex interactions that contribute to variation in fear-related behaviours, such as those displayed in the elevated plus maze and the open field test, although I demonstrate that the contributions are generally small. Also using Bagphenotype, I identify hundreds of gene-by-environment interactions related to these traits. The interacting environmental covariates are diverse, ranging from experimenter to season of the year. With gene expression data from a brain structure associated with anxiety called the hippocampus, I generate modules of co-expressed genes and map them to the genome. Two of these modules were enriched for key nervous system components — one for dendritic spines, another for oligodendrocyte markers — but I was unable to find significant correlations between them and fear-related behaviours. Finally, I employed another Bayesian technique, Sparse Instrumental Variables, which takes advantage of conditional probabilities to identify hippocampus genes whose expression appears not just to be associated with variation in fear-related behaviours, but cause variation in those phenotypes.