Equine herpesvirus-1 (EHV-1) can cause acute upper respiratory tract disease, abortion, neonatal death and neurological disease in horses. Rapid, accurate and timely diagnosis of EHV-1 infection in horses is important to curtail the spread of this pathogen. It has been reported that the neuropathogenic phenotype of EHV-1 can result from a single non-synonymous nucleotide substitution at position 2254 (A→G2254) in open reading frame 30 (ORF30). This was the basis for the development of an allelic discrimination, real-time PCR assay to distinguish between potential neuropathogenic and non-neuropathogenic EHV-1 strains. However, PCR analysis of a panel of EHV-1 abortion isolates revealed that other point mutations within ORF30 could produce false negative results with this previously described assay. Therefore, one of the objectives of this dissertation project was to develop a more sensitive and specific allelic discrimination real-time PCR assay for the detection of EHV-1. This was achieved by redesigning the primers and probes targeting ORF30. The new assay was ten times more sensitive than the original assay, with a lower detection limit of 10 infectious virus particles. While mutations within EHV-1’s genome can hinder diagnosis, they can also impact the virulence of the virus. Objective two, therefore, was to determine if sequential cell passage of T953 would induce sufficient attenuation of the EHV-1 genome to produce a low virulence phenotype. Two separate groups of 28 BALB/c mice were inoculated with either the parental strain or passage 135 (T953 P135) of EHV-1 strain T953. The animals were observed for fourteen days, euthanized and their tissues analyzed for the presence of EHV-1. At the conclusion of the fourteen day observation period, all of the mice infected with T953 P135 survived and regained their pre-inoculation body condition. Furthermore, there were significant differences in virus titer and viral DNA concentrations between T953 P135 and the parental strain, further confirming the attenuated phenotype of the virus. Taken together, data from this study clearly demonstrates that sequential cell culture passage of the neuropathogenic T953 strain of EHV-1 results in attenuation for young adult BALB/C mice.

Os objetivos deste trabalho são avaliar o papel da atenção na percepção de tempo em jovens, idosos e pacientes com a doença de Alzheimer (DA). Nosso trabalho experimental consiste em 4 experimentos. Os experimentos 1 e 2 avaliaram a percepção de tempo em jovens, empregando tarefas de reprodução e de discriminação de durações temporais, respectivamente. Os resultados mostram um efeito atencional nas durações curtas com menor d. Os experimentos 3 e 4 compararam a percepção de tempo entre jovens, idosos e pacientes com DA, empregando tarefas de reprodução e de discriminação de tempo, respectivamente. Os resultados mostram um pior desempenho de idosos e pacientes com DA em relação aos jovens, que parece corresponder a uma deficiência atencional. Concluímos que a modulação da atenção apresenta papel fundamental e crítico na percepção de tempo e os déficits na percepção de tempo em idosos e em pacientes com DA parecem estar associada a uma menor quantidade de recursos atencionais ou a uma dificuldade de focar a atenção. / Our work focused on the attentional modulation of the perception of time. Additionally, other objective was to study the effect of aging and Alzheimers disease (DA) on temporal perception. We have performed four experiments. The first and the second experiments evaluated time perception in young volunteers, by means of the reproduction and discrimination of time intervals, respectively. The results showed an effect of attention on short durations, confirmed by d. The third and fourth experiments had the objective to compare time perception among young volunteers, elderly and DA patients, utilizing reproduction and discrimination of time intervals, respectively. Our findings showed a worse performance in both elderly volunteers and patients with DA in comparison with the group of young volunteers. Our conclusion is that time perception deficits in elderly and patients with DA are associated either to lesser attentional resources or to an increased difficulty in focusing attention on the target, which is more evident in patients suffering of DA.

Os objetivos deste trabalho são avaliar o papel da atenção na percepção de tempo em jovens, idosos e pacientes com a doença de Alzheimer (DA). Nosso trabalho experimental consiste em 4 experimentos. Os experimentos 1 e 2 avaliaram a percepção de tempo em jovens, empregando tarefas de reprodução e de discriminação de durações temporais, respectivamente. Os resultados mostram um efeito atencional nas durações curtas com menor d. Os experimentos 3 e 4 compararam a percepção de tempo entre jovens, idosos e pacientes com DA, empregando tarefas de reprodução e de discriminação de tempo, respectivamente. Os resultados mostram um pior desempenho de idosos e pacientes com DA em relação aos jovens, que parece corresponder a uma deficiência atencional. Concluímos que a modulação da atenção apresenta papel fundamental e crítico na percepção de tempo e os déficits na percepção de tempo em idosos e em pacientes com DA parecem estar associada a uma menor quantidade de recursos atencionais ou a uma dificuldade de focar a atenção. / Our work focused on the attentional modulation of the perception of time. Additionally, other objective was to study the effect of aging and Alzheimers disease (DA) on temporal perception. We have performed four experiments. The first and the second experiments evaluated time perception in young volunteers, by means of the reproduction and discrimination of time intervals, respectively. The results showed an effect of attention on short durations, confirmed by d. The third and fourth experiments had the objective to compare time perception among young volunteers, elderly and DA patients, utilizing reproduction and discrimination of time intervals, respectively. Our findings showed a worse performance in both elderly volunteers and patients with DA in comparison with the group of young volunteers. Our conclusion is that time perception deficits in elderly and patients with DA are associated either to lesser attentional resources or to an increased difficulty in focusing attention on the target, which is more evident in patients suffering of DA.