Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-06-18T14:44:33Z No. of bitstreams: 1 Carlos Adriano de Santana Leal.pdf: 1268535 bytes, checksum: 8c99ca6fceca2937628813239c097e09 (MD5) / Made available in DSpace on 2018-06-18T14:44:33Z (GMT). No. of bitstreams: 1 Carlos Adriano de Santana Leal.pdf: 1268535 bytes, checksum: 8c99ca6fceca2937628813239c097e09 (MD5) Previous issue date: 2017-02-16 / The aim of this study was to standardize a multiplex PCR (mPCR) reaction to detect Microsporum canis, Microsporum gypseum and the Trichophyton mentagrophytes complex in fur and/or crusts samples of dogs and cats. 250 samples were analyzed by direct examination and culture; The DNA from these samples was extracted using the DNeasy Blood & Tissue extraction kit (QIAGEN®, Hilden-Germany). For the PCR, primers were designed for the M. canis, M. gypseum and T. mentagrophytes species and the DNA extracted from colonies of M. canis (URM 6273), M. gypseum (URM 6921) and T. mentagrophytes (URM 6211) from the Collection of Cultures of the Micoteca of the Mycology Department of the Biological Sciences Center of the Federal University of Pernambuco, were utilized as positive controls. A PCR for the detection of M. canis and an mPCR for the detection of M. canis, M. gypseum and the T. mentagrophytes complex was standardized. The protocols standardized in this study, from drawn primers, showed good sensitivity and high specificity in the detection of M. canis, M. gypseum and T. mentagrophytes directly from samples of fur and/or crusts of dogs and cats, making possible a faster and specificity in the results, can be used in the laboratory routine as methods capable of speeding the detection of the agents in question. / Objetivou-se neste estudo padronizar uma reação do tipo multiplex PCR (mPCR) para detectar Microsporum canis, Microsporum gypseum e o complexo Trichophyton mentagrophytes em amostras de pelos e/ou crostas de cães e gatos. Foram analisadas 250 amostras por meio de exame direto e cultura; o DNA destas mesmas amostras foi extraído utilizando-se o kit de extração DNeasy Blood & Tissue (QIAGEN®, Hilden - Germany). Para a PCR foram desenhados primers para as espécies M. canis, M. gypseum e T. mentagrophytes e como controle positivo da reação utilizou-se o DNA extraído de colônias de M. canis (URM 6273), M. gypseum (URM 6921) e T. mentagrophytes (URM 6211), provenientes da Coleção de Culturas da Micoteca do Departamento de Micologia do Centro de Ciências Biológicas da Universidade Federal de Pernambuco. Padronizou-se uma PCR para detecção de M. canis e uma mPCR para detecção de M. canis, M. gypseum e o complexo T. mentagrophytes. Os protocolos padronizados neste estudo, a partir de primers desenhados, apresentaram boa sensibilidade e alta especificidade na detecção de M. canis, M. gypseum e T. mentagrophytes diretamente de amostras de pelos e/ou crostas de cães e gatos, viabilizando um diagnóstico mais rápido e específico, podendo ser empregados na rotina laboratorial como métodos para agilizar a detecção dos agentes estudados.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-06-26T18:12:15Z No. of bitstreams: 1 2016 - Fabr?cia Ferreira e Ferreira.pdf: 1846980 bytes, checksum: c127e17162c67e64b719801f92b0c6fc (MD5) / Made available in DSpace on 2017-06-26T18:12:15Z (GMT). No. of bitstreams: 1 2016 - Fabr?cia Ferreira e Ferreira.pdf: 1846980 bytes, checksum: c127e17162c67e64b719801f92b0c6fc (MD5) Previous issue date: 2016-03-23 / Canine demodicosis is an inflammatory skin disease, frequently diagnosed in veterinary clinics, caused by the proliferation of mites of the species Demodex sp. In recent years, important findings about the disease have been reported, mainly aspects related to treatment, with the insertion of new molecules or new treatment regimens. Doramectin is a macrocyclic lactone that has been used empirically by veterinarians, who use different routes, doses and intervals in its administration, with no homogeneus results. This study aimed to evaluate the use of doramectin in the treatment of dogs affected by the generalized form of demodicosis. Of the forty-six dogs diagnosed with the disease during the study, 20 were selected for the study and divided into three groups: Group I ? treated with doramectin at a dose of 600 mcg/kg once a week orally, group II ? treated at a dose of 300 mcg/kg orally every 3 days and group III ? treated at a dose of 600 mcg/kg every 7 days subcutaneously. The animals were treated until three consecutive negative skin scrapings were obtained, with intervals of at least fifteen days between them (parasitological cure). The days required to obtain the parasitological cure were 105, 82 and 100 according to the indicated groups; and their treatment efficiencies were 75, 100 and 83%, respectively. Doramectin was effective in treating generalized demodectic mange in dogs, regardless of the dose, route and interval of administration. However, the best results were obtained in the group treated at a dose of 300 mcg/kg orally every 3 days. There were no reported adverse reactions with the use of macrocyclic lactone. / Demodiciose canina ? uma doen?a inflamat?ria da pele, frequentemente diagnosticada nos consult?rios veterin?rios, causada pela prolifera??o de ?caros da esp?cie Demodex sp. Nos ?ltimos anos, importantes descobertas sobre a doen?a foram reportadas, principalmente os aspectos relacionados ao tratamento, com a inser??o de novas mol?culas ou novos esquemas de tratamento. A doramectina ? uma lactona macroc?clica que vem sendo usada de forma emp?rica por m?dicos veterin?rios, que a utilizam por diferentes vias, doses e intervalos na sua administra??o, com resultados heterog?neos. O objetivo do estudo foi avaliar a utiliza??o da doramectina no tratamento da demodiciose generalizada em c?es. Dos 46 animais diagnosticados com a doen?a, 20 foram selecionados e divididos em tr?s grupos experimentais: grupo I ? tratado com doramectina dose de 600 mcg/kg semanalmente por via oral, grupo II ? tratado na dose de 300 mcg/ kg por via oral a cada 3 dias e o grupo III ? tratado na dose de 600 mcg/kg a cada 7 dias por via subcut?nea. Os animais foram tratados at? a obten??o de tr?s raspados negativos consecutivos com pelo menos 15 dias de intervalo entre eles (cura parasitol?gica). Os dias necess?rios para obten??o da cura parasitol?gica foram 105, 82 e 100 de acordo com os grupos assinalados e as respectivas efic?cias ao tratamento foram 75, 100 e 83%. A doramectina demonstrou ser eficaz no tratamento da demodiciose generalizada em c?es independente da dose, via e intervalo de sua administra??o. Entretanto, os melhores resultados obtidos foram observados no grupo tratado com a dose de 300 mcg/ kg por via oral a cada 3 dias. N?o foram reportadas quaisquer rea??es adversas com a utiliza??o da lactona macroc?clica.

Made available in DSpace on 2014-08-20T14:31:33Z (GMT). No. of bitstreams: 1 dissertacao_fernanda_antunes_brum.pdf: 139752 bytes, checksum: 989f0c94f1c779f13cc72237d9ab8552 (MD5) Previous issue date: 2010-12-23 / The Ehrlichia canis is a canine monocytic ehrlichiosis responsible for (EMC). The incubation period of EMC is 8 to 20 days, the disease has three phases: acute, subclinical and chronic. The species E. canis is transmitted to the dog and the man by the tick Rhipicephalus sanguineus. It is diagnosed by blood smear, serological or polymerase chain reaction (PCR), with the immunofluorescence method used. Recently E. canis was described as being capable of causing severe disease in humans, with death cases, mainly children and elderly. The gp 19 protein is an important immunodominant antigen, it induces rapid immune response in dogs. The similarity between the geographically distinct samples suggests that the gp19 protein can be used for testing the diagnostic immunoassays, as well as vaccination programs, because this protein is specific for E. canis and thus not have cross reactions with other genera of Ehrlichia.Este study aimed to clone and express the glycoprotein 19 of Ehrlichia canis in Escherichia coli for use as immunobiological, rapid and accurate detection of this disease. The gp 19 gene was amplified by polymerase chain reaction (PCR) using specific primers containing sites for restriction enzymes. The PCR product after digestion, was purified and cloned into a vector and inserted into E. pAE coli TOP 10 competent by electroporation. Of the identified clones was extracted plasmid, which was digested with restriction enzymes to confirm the presence of the insert. After selection of recombinant clones containing the gene linked to the vector, this was purified by heat shock and inserted in expression strain E. coli Star, cultured and induced to express the protein gp19. / A Ehrlichia canis é a responsável pela erliquiose monocitica canina (EMC). O período de incubação da EMC é de 8 a 20 dias; a doença apresenta três fases: aguda, subclínica e crônica. As espécies de E. canis são transmitidas para o cão e para o homem pelo carrapato da espécie Rhipicephalus sanguineus. O diagnóstico é realizado através de esfregaços sanguíneos, métodos sorológicos ou reação em cadeia da polimerase (PCR), sendo a imunofluorescencia o método mais utilizado. Recentemente E. canis, foi descrita como sendo capaz de causar doença grave em humanos, com casos de óbito principalmente em crianças e idosos. A proteína gp 19 é um importante antígeno imunodominante, pois induz rápida resposta imunológica nos cães. A similaridade entre as amostras geograficamente distintas sugere que a proteína gp19 possa ser usada para ensaios de imunoenzimáticos de diagnóstico, bem como em programas vacinais, pois esta proteína é especifica para E. canis não tendo assim reações cruzadas com outros gêneros de Ehrlichia. O gene gp 19 foi amplificado pela reação da polimerase em cadeia (PCR) utilizando oligonucleotídeos iniciadores específicos contendo sítios para enzimas de restrição. O produto da PCR, após digestão, foi purificado e clonado no vetor pAE e inserido em E. coli TOP 10 competente por eletroporação. Dos clones identificados extraiu-se o plasmídio, o qual foi digerido com as enzimas de restrição para comprovar a presença do inserto. Após seleção dos clones recombinantes contendo o gene ligado ao vetor, este foi purificado e inserido por choque térmico na cepa de expressão E. coli Star cultivado e induzido para expressar a proteína gp19. Esta expressão foi verificada por gel de SDS-Page 12% e confirmada por Dot-Blot.

Made available in DSpace on 2014-08-20T13:32:51Z (GMT). No. of bitstreams: 1 dissertacao_elizandra_schoenardie.pdf: 268602 bytes, checksum: 08408f44b3e993faf7cf221741aa401e (MD5) Previous issue date: 2005-06-24 / The Visceral Larva Migrans (VLM) is a zoonotic disease caused by the helminth Toxocara cannis. The precocious diagnosis of this disease in humans is very important to determinate the evolution of the clinical case and the patient's treatment. The goal of the first experiment was determinate the presence of antibodies anti-T. cannis in children from Pelotas through the Indirect Enzyme-Linked Immunosorbent Assay (ELISA) front to the antigen TES, as well as to define the pattern of bands recognized by the positive serums in ELISA through the "Western blotting . For this experiment 427 serums from children, with ages between one to eleven yeas old was tested, those serums was adsorved with AgSoAl and determinate that 50,6% was positive for antibodies anti-TES, showing a significant association among the positive children for antibodies anti-TES and the contact with dogs and cats. This association was also observed in the children's different age groups, but not regarding the gender of the same ones. To perform the Western Blotting , 70 serums witch give a positive result in the Indirect ELISA was been used, all serums recognize glicoproteic bands in the range between 30 and 120 kDa. Was observed a diminution in the crusade reaction with AgSoAl when the adsorved and not adsorved serums with this antigen has been test in the Western blotting , where a band of 30 kDa demonstrate to be an important glicoprotein to specific diagnosis of VLM. In the second experiment, 25 mice BALB/c were inoculated with approximately 1000 eggs containing the larvae L3. Each fifteen days blood collection was made through the reto orbital plexus until the 105 days after de animals infection. The serums was tested in the Indirect ELISA using the Antigen TES and urea 6M in order to discriminate recent and late infection, through the percentile of avidity of the IgG in the different days after the infection. A low percentile of avidity was observed to the 15 days after inoculation (between 7,25 and 27,5%). After 60 days of infection, all the animals presented avidity between 31,4 and 58%. This result suggests that in mice BALB/c, to the 60 days after infection the chronic phase of VLM is already established. / A Larva Migrans Visceral (LMV) é uma doença zoonótica que possui como principal agente etiológico o helminto Toxocara canis. O diagnóstico precoce da doença no homem é importante para estudos de evolução clínica e tratamento do paciente e os inquéritos epidemiológicos para determinar a freqüência da infecção em uma população. Por isso, o primeiro experimento teve como objetivo, determinar a presença de anticorpos anti-T. canis em crianças da região de Pelotas através de Enzyme linked immnosorbent assay (ELISA) com o antígeno de excreção e secreção de larvas de Toxocara canis (TES), bem como definir, através de Western blotting , o padrão de bandas do TES reconhecidas pelos soros positivos ao ELISA. Foram ensaiados no ELISA Indireto 427 soros de crianças de um a 12 anos de idade adsorvidos com antígeno somático de Ascaris lumbricoides e determinado que 50,6% apresentaram anticorpos anti-TES, ocorrendo uma associação significativa entre as crianças positivas e o contato com cães e gatos. Esta associação também foi observada em diferentes faixas etárias das crianças, mas não com relação ao sexo das mesmas. Setenta soros positivos no ELISA foram ensaiados no Western blotting e todos reconheceram frações proteicas entre 30 e 120 kDa. Uma diminuição da reação cruzada com o AgSoAl foi observada quando soros adsorvidos com este antígeno foram testados no Western blotting , sendo que uma fração antigênica de 30 kDa apresentou-se como uma proteína importante para o diagnóstico específico da LMV. No segundo experimento, 25 camundongos BALB/c foram inoculados com aproximadamente 1000 ovos contendo a larva infectante (L3). Colheitas quinzenais de sangue foram realizadas através do plexo retro orbital até os 105 dias pós-infecção dos animais. Os soros foram ensaiados no ELISA Indireto utilizando o antígeno TES e a uréia 6M a fim de discriminar infecção recente e tardia, através do percentual de avidez da IgG nos diferentes dias após a infecção. Um baixo percentual de avidez, característico da infecção aguda, foi observado aos 15 dias pós-inoculação (entre 7,3 e 27,5%). Após 60 dias de infecção, todos os animais apresentaram avidez entre 31,4 e 58%. Através destes resultados, sugere-se que em camundongos BALB/c, aos 60 dias pós-infecção a fase crônica da LMV já está estabelecida.

This study attempted to characterise 5 field stocks of B. canis, collected from various geographic areas of southern Africa in terms of pathogenicity, immune response, cross resistance and antigenic composition. This was done by means of artificial infection of two to four year old Beagle dogs. Serological tests included indirect fluorescent antibody tests, polyacrylamide gel electrophoresis , Western blotting of B. canis antigen and immune serum transfers. The ZIM1, GHT1 , QTN1 and DBN1 stocks were found to be virulent and the JHB1 stock, which might be a cardiac form of babesiosis, was found to be highly virulent. Immune responses of dogs to primary challenges were poor. A second challenge or relapse was important in the development of a resistant immunity. Heterologous challenges showed that dogs which were immune to the ZIM1, QTN1 and DBN1 stocks were partially immune to the JHB1 stock. The dog that was immune to the GHT1 stock was the only one that was totally resistant to the JHB1 stock. Western blots revealed interstock antigenic variation between the JHB1 and the other four stocks. These differences were, however, not great enough to separate the stocks taxonomically.

Context. At the end of their lives, most stars lose a significant amount of mass through a stellar wind. The specific physical and chemical circumstances that lead to the onset of the stellar wind for cool luminous stars are not yet understood. Complex geometrical morphologies in the circumstellar envelopes prove that various dynamical and chemical processes are interlocked and that their relative contributions are not easy to disentangle. Aims. We aim to study the inner-wind structure (R < 250 R-star) of the well-known red supergiant VY CMa, the archetype for the class of luminous red supergiant stars experiencing high mass loss. Specifically, the objective is to unravel the density structure in the inner envelope and to examine the chemical interaction between gas and dust species. Methods. We analyse high spatial resolution (similar to 0 ''.024 x 0 ''.13) ALMA science verification (SV) data in band 7, in which four thermal emission lines of gaseous sodium chloride (NaCl) are present at high signal-to-noise ratio. Results. For the first time, the NaCl emission in the inner wind region of VY CMa is spatially resolved. The ALMA observations reveal the contribution of up to four different spatial regions. The NaCl emission pattern is different compared to the dust continuum and TiO2 emission already analysed from the ALMA SV data. The emission can be reconciled with an axisymmetric geometry, where the lower density polar/rotation axis has a position angle of similar to 50 degrees measured from north to east. However, this picture cannot capture the full morphological diversity, and discrete mass ejection events need to be invoked to explain localized higher-density regions. The velocity traced by the gaseous NaCl line profiles is significantly lower than the average wind terminal velocity, and much slower than some of the fastest mass ejections, signalling a wide range of characteristic speeds for the mass loss. Gaseous NaCl is detected far beyond the main dust condensation region. Realising the refractory nature of this metal halide, this hints at a chemical process that prevents all NaCl from condensing onto dust grains. We show that in the case of the ratio of the surface binding temperature to the grain temperature being similar to 50, only some 10% of NaCl remains in gaseous form while, for lower values of this ratio, thermal desorption efficiently evaporates NaCl. Photodesorption by stellar photons does not seem to be a viable explanation for the detection of gaseous NaCl at 220 R-star from the central star, so instead, we propose shock-induced sputtering driven by localized mass ejection events as an alternative. Conclusions. The analysis of the NaCl lines demonstrates the capabilities of ALMA to decode the geometric morphologies and chemical pathways prevailing in the winds of evolved stars. These early ALMA results prove that the envelopes surrounding evolved stars are far from homogeneous, and that a variety of dynamical and chemical processes dictate the wind structure.

Increased snowmobile use in mountainous terrain has been highlighted as a conservation concern for some Canada lynx (Lynx canadensis) populations. Snow compaction resulting from winter recreation may potentially facilitate access by coyotes (Canis latrans) to habitats used by lynx during winter. Increased interactions could result in either exploitation or interference competition between the two species. Two recent, yet geographically distinct, studies showed contrasting findings regarding coyote movements and their use of snow-compacted trails during the winter. These findings suggest coyote association with snow-compacted trails may be regionally specific and dependent upon ecosystem dynamics and snow characteristics. The objectives of this study were to document diet, space use, and movements of coyotes occupying deep snow regions and explore whether a potential existed for increased interactions between coyotes and lynx due to snowmobile activity. We documented seasonal variation in coyote diets using scat collections to assess dietary overlap with lynx. Coyote resource use within and among habitats containing snowmobile activity was examined using coyote backtrack surveys during two consecutive field seasons in northwestern Wyoming. Although scat analysis findings suggest dietary overlap was not significant between coyotes and lynx during the winter or overall (all seasons combined), we lacked adequate sample size of lynx scats to determine if dietary overlap occurred during the fall, when coyote use of snowshoe hare peaked (24.1 % of all fall occurrences). Coyote backtrack surveys revealed that coyotes not only persisted in habitats used by lynx throughout the winter, but that snow compaction resulting from winter recreation use appeared to influence coyote movements during the winter months. Microhabitat analysis revealed that snow conditions influenced coyote behaviors and habitat use. This research provided insight into the impacts of winter recreation on coyote diet and habitat use during the winter months in northwestern Wyoming. In addition, these results have implications for local lynx populations in the southern periphery of their natural range. These results may assist land management agencies in planning and implementing management strategies to enhance lynx recovery, and may be used to guide decisions regarding areas designated for winter recreation and areas proposed for expansion of winter activities.

While antipredator strategies have been a focus of behavioral research for decades, scientists generally study the responses of prey toward overt, explicit threat. However, risk can also be significant when a threat is covert, such as when an ambush predator may be nearby or a secondary threat remains after a predator's departure. Little is known about the mechanism that prey use to assess risk in a predator's absence. Tests were conducted to determine the manner in which coyotes respond to these ambiguous threats. Specifically, I tested whether coyotes respond to prior anthropogenic activity that has occurred near their only food source, whether they investigate human activity at both profitable (feeding) and unprofitable (non-feeding) locations, and what sort of information coyotes are capable of gaining through their investigation. I explored these questions in three experiments spanning 4 years at the USDA/APHIS/WS National Wildlife Research Center's Logan field station. Test subjects were eight pairs of captive coyotes. Results showed that coyotes delayed or avoided feeding in response to prior anthropogenic activity, and that often a delay was due to investigation of human scent trails. Investigation of non-feeding areas occurred but was relatively brief. When coyotes were prevented from investigating locations of prior anthropogenic activity, foraging ceased altogether. In addition, coyotes were able to differentiate among the activity of different humans based on their association with negative, neutral, or positive threat levels, even in the presence of confounding visual and olfactory cues. They remembered these associations even after one month. This study is the first that provides evidence suggesting that canids gather and interpret complex information for cognitive inference about threat level associated with access to food.

Historically, coyotes were associated with the western United States. During their expansion eastward, coyotes have become more tolerant of humans and have been able to live in varying degrees of urbanization. One main question ecologists around the country are asking is how coyotes are surviving in anthropogenic environments. To aid in answering this question, I have compared coyote land use preference generally and specifically during coyote breeding season, winter and summer, human tourist seasons, and day and night. I also compared coyote land cover preference for deciduous and evergreen cover types during natural seasons. I found that, in general, there was a high variation of preference between and within land use categories. More broadly however, they prefer natural areas over non-natural areas. They used natural and non-natural land use types equally in winter and summer, and during tourist and off-tourist seasons with increased variation in preference during seasons with higher human activity. They had a higher preference for non-natural land use types at night. There is no difference in coyote preference for deciduous or evergreen cover types during the seasons.

No description available.