Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-05-03T14:32:38Z No. of bitstreams: 1 Neurisvan Ramos Guerra.pdf: 2637002 bytes, checksum: 451efc2cbd7dd76dda9b010b8b0189ba (MD5) / Made available in DSpace on 2018-05-03T14:32:39Z (GMT). No. of bitstreams: 1 Neurisvan Ramos Guerra.pdf: 2637002 bytes, checksum: 451efc2cbd7dd76dda9b010b8b0189ba (MD5) Previous issue date: 2017-02-20 / The equid industry in Brazil occupies a prominent position worldwide, with about eight million equids. Diseases caused by protozoa such as Babesia caballi, Theileria equi and Neospora spp. as well as parasites that cause zoonotic protozooses such as Leishmania spp. and Toxoplasma gondii represent one of the main obstacles in the development of the sector. Therefore, this study aims to detect infection by Leishmania spp., Babesia caballi, Theileria equi, Toxoplasma gondii and Neospora spp. and their respective risk factors in equidae created with different management forms. To perform the tests, 400 samples of whole blood and serum from clinically healthy equines, including horses, mules and donkeys from 41 rural properties in the state of Pernambuco were analyzed. In order to detect Leishmania spp., direct parasitological and Polymerase Chain Reaction (PCR) tests were performed. Concerning the presence of infection by Babesia caballi and Theileria equi, direct parasitological tests and enzyme-linked immunosorbent assay (ELISA) were used for anti-Babesia caballi and anti-Theileria equi immunoglobulins detection. For the determination of seroprevalences of toxoplasmosis and neosporosis, modified agglutination (MAT) tests were used to identify anti-T. gondii IgG antibodies and anti-Neospora spp. All samples were negative for Leishmania spp. in the tests, suggesting that equidae do not participate in the epidemiological chain of leishmaniasis in the studied areas. The prevalence of anti-Babesia caballi and anti-Theileria equi antibodies of 4.3% (17/400; CI: 2.6-6.9) and the presence of B. caballi and T. equi in the serological tests revealed a prevalence of 10.8% (43/400; CI: 8.0 - 14.3), respectively, and co-infection was detected in 1% (4/400) of the animals. These data allow the characterization of areas of enzootic instability in the sites surveyed. Anti-T. gondii IgG antibodies were detected in 12.5% (50/400) of the animals analyzed. When evaluating the risk factors for T. gondii infection, only the mesoregion factor (p = 0.029) was associated with infection, particularly Zona da Mata (OR = 3). The results reveal the presence of the parasite in the studied area, which may represent a link in the transmission chain of toxoplasmosis. Seropositivity for Neospora spp. was 5.7% (23/400) and the variables age, breeding type and region presented statistical significance. In relation to age, it was observed that animals older than 11 years presented 11.8 times more chances of being serum-reactive wjhen compared with young animals (<2,5) and the prevalence found shows that the parasite is dispersed in the areas studied and that the variables age, breeding type and region are the most important risk factors for the occurrence of infection in equidae, and should be considered in the prevention of the disease. Considering the results found in the present study, the diagnosis of the various diseases present in the State of Pernambuco, when performed at an early stage, allows the application of preventive and control measures, contributing significantly to animal health and public health. / A equideocultura do Brasil ocupa posição de destaque mundial, com cerca de oito milhões de cabeças. Doenças causadas por protozoários como Babesia caballi, Theileria equi e Neospora spp. além de parasitos que causam protozooses zoonóticas a exemplo de Leishmania spp. e Toxoplasma gondii representam um dos principais entraves no desenvolvimento do setor. Diante disso, esse estudo tem como objetivo determinar as prevalências e fatores de risco associados às infecções por Leishmania spp., Babesia caballi (Nuttall & Strickland, 1910), Theileria equi (Mehlhorn & Schein, 1998), Toxoplasma gondii (Nicolle & Manceaux, 1909) e Neospora spp. em equídeos submetidos a diferentes regimes de criação. Para realização dos exames, foram analisadas 400 amostras de sangue total e soro de equídeos clinicamente saudáveis, incluindo equinos (387/400), muares (9/400) e asininos (4/400) provenientes de 41 propriedades rurais do estado de Pernambuco. Com a finalidade da detecção de Leishmania spp., foram realizados os exames parasitológicos diretos e Reação em cadeia da Polimerase (PCR). No intuito de averiguar a presença de infecção por Babesia caballi e Theileria equi foram utilizados os exames parasitológico direto e Ensaio de Imunoadsorção Enzimática (ELISA), para detecção de imunoglobulinas anti-Babesia caballi e anti-Theileria equi. Para determinação das soroprevalências da toxoplasmose e neosporose foram utilizados os testes de aglutinação modificado (MAT) para identificação de anticorpos IgG anti-T. gondii e IgG anti-Neospora spp. Todas as amostras resultaram negativas para Leishmania spp. nos testes, o que sugere que os equídeos não participam da cadeia epidemiológica das leishmanioses nas áreas estudadas. Quanto à presença de B. caballi e T. equi, os testes sorológicos revelaram uma prevalência de anticorpos anti-Babesia caballi e anti-Theileria equi de 4.3% (17/400; I.C. 2,6 – 6.9%) e 10,8% (43/400; I.C. 8.0 – 14.3), respectivamente e foi detectada co-infecção em 1% (4/400) dos animais. Tais dados permitem caracterizar como áreas de instabilidade enzoótica os locais pesquisados. Anticorpos IgG anti-T. gondii foram detectados em 12,5% (50/400) dos animais analisados. Quando avaliados os fatores de risco para infecção por T. gondii, apenas o fator mesorregião (p=0,029) apresentou associação com a infecção, particularmente Zona da Mata (OR=3). Os resultados revelam a presença do parasito na área estudada, o que pode representar um elo na cadeia de transmissão da toxoplasmose. A soropositividade para Neospora spp. total foi de 5,7% (23/400) e as variáveis idade, tipo de criação e região apresentaram significância estatística. Em relação à idade, observou-se que animais acima de 11 anos apresentaram 11,8 vezes de chances a mais de serem sororreagentes quando comparados com os animais jovens (<2,5) e a prevalência encontrada demonstra que o parasito está disperso nas áreas estudadas e que as variáveis idade, tipo de criação e região são fatores de riscos mais importantes para ocorrência da infecção em equídeos, devendo ser considerados na prevenção da doença. Considerando os resultados encontrados no presente estudo, o diagnóstico das diversas doenças presentes no estado de Pernambuco, quando realizado de forma precoce, possibilita a aplicação de medidas preventivas e de controle, contribuindo significativamente com a sanidade animal e saúde pública.

Made available in DSpace on 2014-08-20T14:38:04Z (GMT). No. of bitstreams: 1 dissertacao_alvaro_carlos_menezes_arteche.pdf: 1268541 bytes, checksum: 48cb8939395a534c09b30fdb0ec17ef9 (MD5) Previous issue date: 2011-11-21 / This study analyzed, by reaction to indirect immunofluorescence and individual weighting, the levels of protection and weight-gain in beef calves from birth to the age of eight month, which received two doses of vaccine against tick fever and were maintained free from ticks, compared to naturally infested calves in extensive management conditions with continuous grazing in a rural property situated in the municipality of Santana do Livramento Espinilho / RS. Two groups were randomly gathered, n=30, from which Group I (test) was kept free from ticks from birth to weaning and received two doses of the attenuated, trivalent, refrigerated vaccine against tick fever. Groupe II (control) followed the property s traditional management, which reflects the one used in the region, allowing the tick infestation by the animals. Weight control was accomplished in the first day (d0) and in the last day of the experiment (d180). The serology revealed that 100% of the vaccinated animals presented titles equal to 1:5120 for the three parasites Anaplasma marginale, Babesia bovis e Babesia bigemina (p≤0,025), whereas in the control group the highest titles were 1:2560 in four animals for A. marginale, 1:1280 in four animals for B. bovis and 1:1280 in two animals for B. bigemina (p≤ 0,025). Group I got a mean weight-gain of 30,5 kg more than the control group at the end of the experiment (p≤0,025). The results obtained show that keeping animals free from tick, from birth to weaning, and maintaining them vaccinated against tick fever is safer, more efficient and economically and technically more advantageous than the traditional management system carrapateamento (natural tick infestation). / Avaliaram-se, pela reação de imunofluorescência indireta (RIFI) e de pesagens individuais, os níveis de proteção e ganho de peso de terneiros de corte do nascimento ao desmame com oito meses de idade que receberam duas doses de vacina contra tristeza bovina e foram mantidos livres de carrapatos em comparação a terneiros naturalmente infestados em condições de manejo extensivo com pastoreio contínuo em propriedade rural localizada no município de Santana do Livramento-Espinilho/RS. Formaram-se dois grupos aleatóriamente, n=30, sendo o GRUPO I (teste) mantido livre de carrapatos desde o nascimento até o desmame e com duas doses da vacina atenuada, trivalente e refrigerada contra tristeza bovina. O GRUPO II, controle, seguiu o manejo tradicional da propriedade, que reflete o da região, permitindo que os animais fossem infestados por carrapatos. As pesagens foram realizadas no primeiro dia do experimento (d0) e no último dia do experimento (d180). A sorologia revelou que 100% dos animais vacinados apresentaram títulos maiores ou iguais a 1:5120 para os três parasitos Anaplasma marginale, Babesia bovis e Babesia bigemina (p≤0,025) enquanto que no grupo controle os títulos máximos foram 1:2560 em quatro animais para A. marginale, 1:1280 em quatro animais para B. bovis e 1:1280 em dois animais para B. bigemina. Houve diferença significativa entre os dois grupos (p≤0,025). O grupo I obteve 30,5 Kg a mais de ganho médio de peso que o grupo controle no final do experimento (p≤0,025). Com os resultados obtidos, conclui-se que é mais eficiente, seguro e vantajoso econômica e tecnicamente manter os animais livres de carrapatos, desde o nascimento até o desmame, e vacinados contra tristeza bovina, do que com o manejo tradicional carrapateamento .

Made available in DSpace on 2014-08-20T13:32:57Z (GMT). No. of bitstreams: 1 dissertacao_marcelo_gotze.pdf: 556489 bytes, checksum: cb0ceeb57d3c58da117f1a655a510d0d (MD5) Previous issue date: 2010-03-18 / Bovine babesiosis caused by Babesia bovis and Babesia bigemina, is the most important disease transmitted by Rhipicephalus (Boophilus) microplus in tropical and subtropical areas in South America. Definitive diagnosis can be made by detecting infected erythrocytes in blood smears. However, the parasitemia in peripheral blood is often too low for this method to be used for diagnostic purposes. For this reason, several serological tests, including complement fixation, indirect hemagglutination and indirect immunofluorescence (IIF) have been used to detect antibodies in infected cattle. Although these tests allow the detection of persistently infected animals, they have limitations in specificity and/or sensitivity. The IIF has been the most sensitive, but cross-reactivity between species, subjective interpretation, and low production has limited its usefulness. The enzyme linked immunosorbent assay (ELISA) have found wide application in the diagnosis of infectious diseases. The cELISA format (competitive) can provide an additional level of specificity, because the antibody is directed to a single epitope, specific for the organism to be detected. For these reasons, this study aimed to evaluate the sensitivity and specificity of cELISA compared to IIF and nested PCR (nPCR) for diagnosis babesiosis caused by B. bovis. Therefore, blood samples were collected from cattle in Brazil and Argentina, and processed for the diagnosis of B. bovis. The nPCR was used as the gold standard to validate the cELISA and the IIF as a comparative test. The cELISA for the diagnosis of B. bovis presented is easily processed with high levels of sensitivity and specificity. It is easily performed in a high number of samples, making it useful in cases of outbreaks of bovine babesiosis. / A babesiose bovina, causada por Babesia bovis e Babesia bigemina, é a doença mais importante transmitida por carrapatos Rhipicephalus (Boophilus) microplus em áreas tropicais e subtropicais da América do Sul. O diagnóstico definitivo pode ser feito através da detecção de eritrócitos infectados em esfregaços sanguineos, porém a parasitemia em sangue periférico é frequentemente muito baixa para que esse método seja utilizado de forma confiável para fins de diagnóstico. Por esse motivo, vários testes sorológicos, incluindo a fixação de complemento, hemaglutinação indireta e imunofluorescência indireta (IIF) têm sido usados para detectar anticorpos em bovinos infectados. Embora estes testes permitam a detecção de animais persistentemente infectados, eles têm limitações na especificidade e/ou sensibilidade. O IIF tem sido o mais sensível, mas a reatividade cruzada entre as espécies, interpretação subjetiva, e baixa produção tem limitado a sua utilidade. Os ELISA têm encontrado ampla aplicação no diagnóstico de doenças infecciosas. O formato cELISA (competitivo) pode fornecer um nível adicional de especificidade, pois o anticorpo é dirigido para um epitopo único específico para o organismo a ser detectado. Por estas razões, este estudo teve como objetivo avaliar a sensibilidade e especificidade do cELISA comparado com IIF e nested PCR (nPCR) para diagnóstico de babesiose causada por B. bovis. Para tanto, amostras de sangue bovino foram coletadas no Brasil e na Argentina e processadas para o diagnóstico de B. bovis. Utilizou-se o nPCR como teste padrão para a validação do cELISA, e a IIF como teste comparativo. O cELISA para diagnóstico de B. bovis apresentou-se de fácil processamento, com altos níveis sensibilidade e especificidade, além da rapidez no processamento de amostras em larga escala, sendo de grande utilidade para casos de surtos de babesiose bovina

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.

This observational study of 100 dogs naturally infected with Babesia rossi determined whether severity of parasitaemia was associated with outcome of infection and documented the relative distribution of parasitised red blood cells (pRBC) in capillary and venous circulation. The association between increased parasitaemias and outcome with a clinically compromised circulation was also investigated. Outcome was defined as either hospitalisation with death, or hospitalisation with eventual recovery or treatment as an outpatient. Dogs were enrolled if large babesias were found on stained thin capillary blood smears made from an ear prick. Thin venous smears were prepared from jugular or cephalic blood. Parasitaemias were manually counted and expressed as the percent pRBC. Ten dogs died, 50 recovered after hospitalisation and 40 were treated as outpatients. Venous sampling site did not affect venous parasitaemia (P = 0.6). Both capillary and venous parasitaemias of dogs that died were significantly higher than those of dogs that recovered after hospitalisation (P = 0.002) and dogs that were treated as outpatients (P < 0.0001). When assessing the whole group, capillary parasitaemia (median 0.61%, range <0.05-71.6%, interquartile range (IQR) 0.22-3.75%) was significantly higher than venous parasitaemia (median 0.14%, range 0-30.6%, IQR 0.046–0.52%) with P < 0.0001. The 21 dogs with a clinically compromised circulation were more likely to die (P <0.0001) and had significantly higher capillary (median 5.98%, range 0.09-71.6%, IQR 2.44-19.41%) and venous (median 2.81%, range <0.05-30.6%, IQR 0.17-9.03%) parasitaemias than the 79 dogs with a clinically normal circulation (capillary median parasitaemia 0.38%, range <0.05-12.87%, IQR 0.16-1.42%; venous median parasitaemia 0.096%, range 0-6.13%, IQR <0.05-0.33%; P < 0.0001). This study shows that high parasitaemia is significantly associated with death in B rossi infected dogs. Unfortunately, there was a wide overlap in the parasitaemias of the three outcome groups with the result that neither capillary nor venous parasitaemias appear prognostically useful. The previous clinical suspicion that capillary parasitaemias are usually higher than venous parasitaemias is confirmed. Thus capillary samples are the most appropriate diagnostic samples. Prior observations that a clinically compromised circulation is associated with death are confirmed. This association provides a rapid means of identifying patients in need of intensive monitoring and treatment. Despite the highly significant association between compromised circulation and higher parasitaemia, it is thought unlikely that parasite burden is the sole trigger for circulatory collapse. Copyright 2006, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Bohm, M 2006, The comparative assessment of capillary and venous Babesia rossi parasitaemias on thin blood smears and their association with disease manifestation, MMedVet dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-05042007-154527 / > / Dissertation (Master of Veterinary Medicine (Small Animal Medicine))--University of Pretoria, 2006. / Companion Animal Clinical Studies / unrestricted

Babesia bovis outbreaks were reported in cattle alleged to be immunised with the commercially available live-blood vaccine in the Swartberg region of KwaZulu-Natal, South Africa and an investigation into the nature of parasites causing the outbreaks was carried out. The H isolate was obtained from a clinically ill animal on the Haistings farm and characterised using BvVA1 and Bv80 size analysis coupled with Bv80 and 18S rRNA V4 hypervariable region sequence analysis. In total, four South African B. bovis isolates were analysed: the vaccine stock (S) at passage 11 and 23 and field isolates H and F. The S23 strain used to infect vaccine donor animals could not be detected in the H isolate and could not be responsible for the severe disease symptoms observed in the field animals. Sequence profiles of the Bv80 and 18S rRNA V4 hypervariable regions for all detectable strains were compiled and now serve as a basis for the investigation of future babesiosis outbreaks. It was determined that the Bv80 PCR is not able to detect animals at the carrier stage of infection and that non-specific primer binding to Boophilus microplus and Boophilus decoloratus tick DNA occurs. For this reason, the Bv80 PCR is not suitable for investigating the nature of B. bovis infections in ticks. The BvVA1 PCR reaction required extensive optimisation and did not detect all strains present in the isolates and was therefore not used as a basis for strain discrimination. Microaerophilous stationary phase cultures of the vaccine strain at passage 24 (S24) and the H strain were initiated as a potential source of soluble parasite antigens. Continuous cultivation was not possible despite the alteration of a number of conditions. Currently there is no culture adapted B. bovis strain in South Africa and the availability of such a strain would form the basis of studies on the development of alternative vaccines. / Dissertation (MSc)--University of Pretoria, 2008. / Veterinary Tropical Diseases / unrestricted

As part of the larger survey to map the geographical distribution of Babesia and Theileria parasites in the Southern African rhinoceros population, white rhinoceroses were sampled during routine immobilizations in the Kruger National Park. Polymerase Chain Reaction (PCR) and Reverse Line Blot (RLB) hybridization assays were used to screen for the presence of haemoprotozoa and complete blood counts were used to assess associated changes in clinical parameters. Of the 195 rhinoceroses sampled, 36.4% tested positive for the presence of Theileria bicornis, with no significant change in the haematological parameters measured. None of the rhinoceroses sampled tested positive for Babesia bicornis, the parasite linked to mortalities in black rhinoceroses. Copyright / Dissertation (MSc)--University of Pretoria, 2009. / Veterinary Tropical Diseases / unrestricted

The Affymetrix GeneChip® Canine Genome 2.0 microarray is re-annotated using AgBase tools, up-to-date ID mapping and GO annotations associated with publicly available gene products updated on this array. This re-annotation makes the array more useful for researchers using the canine microarray for biological discovery. We use flow cytometry to determine if liposomal clodronate (LC) is an acceptable alternative to surgical splenectomy to facilitate detection of subclinical infection with Babesia canis in potential blood donor greyhounds. Our study shows that LC is not a reliable means of exposing babesiosis in greyhounds with a recent history of infection. We evaluate the effect of depletion of antigen presenting cells on regulatory T cells (Tregs) in dogs treated with LC by multi-color flow cytometry. We demonstrate that LC promotes increases in the CD4+CD25+FOXP3+ Tregs affecting mostly the CD4+CD25lowFOXP3+ Tregs subset suggesting a role of monocytes in naïve T cell priming and differentiation into Tregs.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-15T11:44:50Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_PrevalenciaSorologicaMolecular.pdf: 861713 bytes, checksum: ffddee7491410356b82fbbbef16f58ea (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-17T12:36:50Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_PrevalenciaSorologicaMolecular.pdf: 861713 bytes, checksum: ffddee7491410356b82fbbbef16f58ea (MD5) / Made available in DSpace on 2017-05-17T12:36:50Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_PrevalenciaSorologicaMolecular.pdf: 861713 bytes, checksum: ffddee7491410356b82fbbbef16f58ea (MD5) Previous issue date: 2014-06-26 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo do presente estudo foi verificar a prevalência sorológica e molecular de Babesia bovis e Babesia bigemina em búfalos da Ilha de Marajó, estado do Pará, Brasil. Foi utilizado o Ensaio de Imunoadsorção Enzimático indireto (ELISAi) com antígeno total contendo proteínas de superfície externa e a Reação em Cadeia da Polimerase quantitativa (PCRq), envolvendo o uso de SYBR Green com base na amplificação de um pequeno fragmento de gene do citocromo b. A prevalência de animais positivos no ELISAi para B. bovis, B. bigemina e para infecção mista foi de 24.87% (199/800), 20.75% (166/800) e 18.75% (150/800), respectivamente. Na PCRq foi detectado a presença de B. bovis em 15% (18/199) e de B. bigemina em 16% (19/199) dos animais, sendo que destes, 58% (11/19) apresentavam-se co-infectados pelos dois agentes. Os resultados mostram uma baixa prevalência de anticorpos anti-B. bovis e anti-B. bigemina em búfalos da Ilha do Marajó. Porém, observou-se que os agentes da babesiose bovina circulam em búfalos, podendo atuar como reservatório. / The aim of the study was to estimate the prevalence of Babesia bovis and Babesia bigemina in water buffaloes of the Marajó Island, State of Pará, Brazil. We used an indirect enzyme-linked immunosorbent assay (iELISA), with total antigen containing proteins outer surface, and polymerase chain reaction (qPCR), involving the use of SYBR Green based on amplification of a small fragment of the cytochrome b gene. The prevalence of positive animals in iELISA to B. bovis B. bigemina and mixed infection was 24.87% (199/800), 20.75% (166/800) and 18.75% (150/800), respectively. Using the PCR, the presence of B. bovis was detected in 15% (18/199) and B. bigemina in 16% (19/199) of animals, and of these, 58% (11/19) presented co-infected by the two agents. The results show a low prevalence of an- tibodies anti-B. bovis and anti-B. bigemina in water buffaloes from Marajó Island. However, it was observed that the agents of bovine babesiosis circulate in buffaloes, and these may act as reservoirs.

No description available.