La anaplasmosis canina es una enfermedad rickettsial zoonótica causada por bacterias intracelulares obligatorias gram negativas siendo Anaplasma phagocytophilum, responsable de la anaplasmosis Granulocítica Canina (AGC) y Anaplasma platys causante de la Trombocitopenia Cíclica Infecciosa Canina (TCIC) y que son transmitidos por la mordedura de garrapatas Ixodes spp. y Rhipicephalus sanguineus, respectivamente. En nuestro país existen pocos reportes de estos agentes en perros, más sí hay evidencia de la garrapata vector. El objetivo de este estudio fue determinar anaplasmosis canina mediante el hallazgo hematológico (evidencia de corpúsculos de inclusión o mórulas más trombocitopenia) y presencia de anticuerpos contra Anaplasma spp. en perros con antecedentes de garrapatas del distrito de Chiclayo (Lambayeque - Perú). Para ello se colectaron 88 muestras sanguíneas de perros aparentemente sanos y con antecedentes de mordeduras por garrapatas. El hallazgo hematológico evidenció que el 2.27% (2/88) de perros fueron positivos (presentaron corpúsculos de inclusión o mórulas más trombocitopenia) resultando ser compatibles con A. platys. Además, se detectó una seropositividad a Anaplasma spp. en un 22.73% (20/88) y una seropositividad múltiple del 21.59% (19/88) con Ehrlichia spp. mediante el Test SNAP 4Dx Plus. Por otro lado, los perros positivos y seropositivos evidenciaron de mayor a menor frecuencia alteraciones hematológicas como trombocitopenia, anemia, leucocitosis y leucopenia. Estos resultados demuestran anaplasmosis canina de tipo subclínico en perros con antecedentes de garrapatas del distrito de Chiclayo (Lambayeque - Perú). / Universidad Nacional Mayor de San Marcos (Lima). Vicerrectorado de Investigación y Posgrado / Universidad Nacional Mayor de San Marcos (Lima). Facultad de Medicina Veterinaria. Laboratorio de Patología Clínica Veterinaria / Tesis

Orientador: Rosangela Zacarias Machado / Resumo: Anaplasma marginale é uma bactéria, Gram negativa, intracelular obrigatória parasita de eritrócitos e o principal agente da Anaplasmose bovina. Esta doença causa anemia severa, provocando a redução do ganho de peso e da produção de leite e gerando grandes perdas econômicas na pecuária mundial. A diversidade genética desta bactéria vem sendo caracterizada com base na sequência das proteínas de superfície (MSPs), principalmente, na proteína MSP1α, sendo possível identificar as diferentes estirpes geográficas de acordo com as diferenças nas sequências de aminoácidos. O presente estudo teve como objetivo investigar a diversidade genética de A. marginale em bovinos de corte da raça Angus naturalmente infectados durante um surto da enfermidade. Vinte bovinos, com idades entre 6 e 9 meses, oriundos de uma fazenda no município de Itú, estado de São Paulo foram acompanhados por quatro meses e quatro colheitas de sangue foram realizadas. Amostras de soro foram submetidas à Ensaio Imunoenzimático Indireto (iELISA) para detecção de anticorpos IgG anti-A. marginale. As oitenta amostras de sangue total obtidas, foram submetidas à extração de DNA, PCR em tempo real quantitativa (qPCR) para o gene msp1β, semi-nested PCR (snPCR) para o gene msp1α, clonagem do fragmento alvo e sequenciamento pelo método de Sanger. As sequências obtidas foram submetidas à analises de diversidade genética pelo software RepeatAnalyzer. O ensaio Imunoenzimático Indireto (iELISA), utilizando Ag total, revelou baixa... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Anaplasma marginale is a Gram-negative, obligatory intracellular erythrocyte parasite and the main agent of bovine Anaplasmosis. This disease causes severe anemia, causing a reduction in weight gain and milk production and generating major economic losses in livestock worldwide. The genetic diversity of this bacterium has been characterized based on the sequence of surface proteins (MSPs), mainly in the protein MSP1α, making it possible to identify the different geographical strains according to the differences in the amino acid sequences. The present study aimed to investigate the genetic diversity of A. marginale in Angus beef cattle naturally infected during an outbreak of the disease. Twenty cattle, aged 6 to 9 months, from a farm in the city of Itú, state of São Paulo, were followed for four months and four blood samples were taken. Serum samples were subjected to the Indirect Immunoenzymatic Assay (iELISA) to detect antibodies IgG anti-A. marginale. The eighty whole blood samples obtained were subjected to DNA extraction, quantitative real-time PCR (qPCR) for the msp1β gene, semi-nested PCR (snPCR) for the msp1α gene, cloning of the target fragment and sequencing by the Sanger method. The obtained sequences were subjected to genetic diversity analysis using the RepeatAnalyzer software. The Indirect Immunoenzymatic assay (iELISA) revealed low seroprevalence in the sampled animals despite 100% positivity in the qPCR, with quantification between 103 and 107 number of DNA c... (Complete abstract click electronic access below) / Mestre

Orientador: Marcos Rogério André / Coorientador: Rosangela Zacarias Machado / Banca: Estevam Guilherme Lux Hoppe / Banca: Darci Moraes Barros-Battesti / Banca: Heitor Miraglia Herrera / Banca: Isabel Kinney Ferreira de Miranda Santos / Resumo: Poucos são os estudos acerca da diversidade genética de Anaplasma marginale nos rebanhos bovinos brasileiros, principalmente no que diz respeito a bovinos de corte. O presente estudo teve como objetivo avaliar a diversidade genética de A. marginale, com base nos genes msp1α e msp4, em Bos taurus indicus amostrados no Pantanal brasileiro. Alíquotas de sangue com e sem EDTA foram colhidas de 400 bovinos (200 vacas e 200 bezerros) em cinco propriedades de cria e recria extensiva. Enquanto as amostras de soro foram submetidas ao Ensaio Imunoenzimático Indireto (iELISA) para detecção de anticorpos IgG anti-A. marginale, amostras de sangue total foram submetidas à avaliação do hematócrito e confecção de esfregaços sanguíneos corados pelo Giemsa, PCR em tempo real quantitativa (qPCR) baseada no gene msp1β, semi-nested (sn)PCR baseada no gene msp1α e PCR convencional (cPCR) baseada no gene msp4 para A. marginale. Os produtos amplificados nos ensaios de snPCR e cPCR foram purificados e sequenciados pelo Método de Sanger. Sequências do gene msp1α foram submetidas à análise de diversidade pelo software RepeatAnalyser. Sequências do gene msp4 foram submetidas à análise de distância Network pelo software Splitstree e de genótipos pelo Software DnaSP5 e PopART. Cinco animais (duas vacas e três bezerros) apresentaram corpúsculos de A. marginale em esfregaços sanguíneos. As frequências de animais positivos pelo iELISA, qPCR e snPCR foram de 72,25% (289/400), 56,75% (227/400) e 22,9% (52/227)... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: There are few studies about the genetic diversity of Anaplasma marginale in Brazilian cattle herds, especially with regard to beef cattle. The present study aimed to evaluate the genetic diversity of A. marginale based on the msp1α and msp4 genes in Bos taurus indicus, sampled in the Brazilian Pantanal. Blood aliquots with and without EDTA were collected from 400 cattle (200 cows and 200 calves) in five extensive breeding and rearing properties. While the serum samples were submitted to the Indirect Immunoenzyme Assay (iELISA) for the detection of anti-A. marginale IgG antibodies, whole blood samples were submitted to the evaluation of hematocrit and Giemsa-stained blood smears, quantitative real-time PCR (qPCR) based on the msp1β gene, semi-nested (sn) PCR based on the msp1α gene and conventional PCR (cPCR) based on the msp4 gene for A. marginale. The amplified products obtained in the snPCR and cPCR assays were purified and sequenced by the Sanger Method. Sequences of the msp1α gene were submitted to diversity analysis by the RepeatAnalyser software. msp4 gene sequences were submitted to Network distance analysis using the Splitstree software and genotypes analysis by the DnaSP5 and PopART software. Five animals (two cows and three calves) presented A. marginale corpuscles in blood smears. The frequencies of iELISA positive animals, qPCR and snPCR were 72.25% (289/400), 56.75% (227/400) and 22.9% (52/227), respectively. The iELISA and qPCR tests showed weak kappa concordance. Cows (154/200) were statistically more seropositive than calves (135/200) (P <0.05). In the qPCR, the number of calves (138/200) and mean value of quantification (1.3 x 106 copies msp1α A. marginale/μL) were higher when compared to those found for cows (89/200; 3.9 x 104) (P <0.05). The rickettsemia estimated by qPCR showed no corr... (Complete abstract click electronic access below) / Doutor

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-11T19:12:48Z No. of bitstreams: 1 2012 - F?bio Jorge Moreira da Silva.pdf: 1417566 bytes, checksum: a168314142bf607a64227972ff4c85e8 (MD5) / Made available in DSpace on 2017-05-11T19:12:48Z (GMT). No. of bitstreams: 1 2012 - F?bio Jorge Moreira da Silva.pdf: 1417566 bytes, checksum: a168314142bf607a64227972ff4c85e8 (MD5) Previous issue date: 2012-06-06 / CNPq / Activities in high productivity systems to dairy cattle contribute to imbalance health and disease, and increase the possibility of illness compatible with the infestation and infections transmitted by arthropods vectors. The objectives were to evaluate the prevalence and the seroepidemiological condition for Anaplasma marginale in 41calves from birth to completed 180 days. The animals belonged to ?Centro Estadual de Pesquisa em Agricultura Org?nica? ? Pesagro-Rio, Serop?dica-RJ. There are few reports of molecular diagnostic techniques for this agent. The study was conducted during the rainy and dry seasons and collected a total of 1607 blood samples, initially every three days and processed using indirect ELISA test and PCR. Percent values for A. marginale seroprevalence as function of age were tested using the ?2 test at 5% significance level. The prevalence of anti-A. marginale antibodies were 39.8% in calves aged less than 30 days, 23.3% between 30 and 60 days, 27.3% between 60 and 120 days and 38.2% between 120 and 180 days, with 31.4% for samples of all age group (180 days). Calves aged between 30 and 60, 60 and 120 and 120 and 180 days were respectively 1.90, 1.75 and 1.55 more likely to be seronegative for A. marginale than newborn ones. All calves were positive to PCR until 13 days old. The values show that during the study calves had low levels of antibodies to A. marginale, a condition that predisposes them to the development of clinical anaplasmosis. In addition, the herd was considered unstable epidemiologically to A. marginale infection. The results show that animals had low antibody titers of IgG anti-A. marginale, being more susceptible to develop clinical anaplasmosis from 30 to 60 days. The results of the PCR method confirmed A. marginale in all animals before they are 15 days old and suggest the possibility of transplacental transmission occurs in the herd. / As atividades relacionadas a bovinocultura leiteira em sistemas de alta produtividade contribuem cada dia mais com o desequil?brio sa?de-doen?a, e aumentam a probabilidade de enfermidades compat?veis com as ectoparasitoses e as infec??es transmitidas por estes vetores. Os objetivos do trabalho foram detectar a infec??o por Anaplasma marginale de forma precoce e avaliar o n?vel de anticorpos IgG anti-A. marginale em bezerros nativos e naturalmente parasitados por Rhipicephalus microplus na Baixada Fluminense, estado do Rio de Janeiro. S?o escassos os relatos sobre t?cnicas de diagn?stico molecular para este agente. Um total de 41 bezerras foi acompanhado do nascimento aos 180 dias de idade. Os animais pertenciam ao Centro Estadual de Pesquisa em Agricultura Org?nica da Pesagro-RJ. O estudo foi conduzido nas esta??es chuvosa e seca, e foram coletados um total de 1607 amostras, com intervalo inicial de tr?s dias e processados utilizando o teste ELISA indireto e a t?cnica PCR. Os valores percentuais de soropreval?cia para A. marginale em fun??o da idade foram submetidos ao teste ?2 a 5% de signific?ncia. A preval?ncia de anticorpos anti-A. marginale nos bezerros, em fun??o da idade, foi de 39,8% do soro de animais com idade inferior a 30 dias, 23,3% entre 30 e 60 dias, 27,3% entre 60 e 120 dias e 38,2% entre 120 e 180 dias, com um percentual de 31,4% para as amostras de todo o grupo et?rio (180 dias). Bezerros com idade 30 - 60, 60 - 120 e 120 - 180 dias apresentaram respectivamente 1,90, 1,75 e 1,55 mais risco de serem soronegativos para A. marginale do que os animais rec?m nascidos. Os bezerros foram diagnosticados positivos a PCR em no m?ximo 13 dias de idade. Os valores demonstram que os animais estudados apresentaram baixos t?tulos de anticorpos da classe IgG anti-A. marginale, sendo mais suscet?veis a desenvolverem anaplasmose cl?nica entre 30 a 60 dias de vida. Os resultados do m?todo PCR comprovaram a circula??o de A. marginale em todos os animais antes de completarem 15 dias de vida e sugerem a possibilidade de ocorrer transmiss?o transplacent?ria no rebanho estudado.

Lyme borreliosis was first described some 30 years ago in the USA. Today, it is the most common vector borne disease in Europe and the USA. The disease can have multiple stages and symptoms can manifest from various parts of the body; joints, skin heart and nervous system. In Europe, neuroborreliosis is the most frequent late stage diagnosis. Although Lyme borreliosis is treatable with antibiotics and the causative spirochete has not been shown to be resistant to drugs, some patients do not recover completely. They have persistent symptoms and are diagnosed with chronic or persistent Lyme borreliosis. The mechanism behind the lingering symptoms is unclear but might be due to tissue damage caused by the immune system. The aim of this thesis was to study the immunological differences between patients with different outcome of Lyme borreliosis, i.e. chronic, subacute and asymptomatic, and various factors that might influence the course of the disease. The Borrelia-specific IFN-γ and IL-4 secretion was detected in blood and cerebrospinal fluid from patients with chronic and subacute neuroborreliosis during the course of the disease. Blood samples were also obtained from patients with erythema migrans (EM) and acrodermatitis chronicum atrophicans. An early increase of IFN-γ with a later switch to an IL-4 response was observed in patients with a subacute disease course whereas the IFN-γ secretion continued to be elevated in chronic patients. The Borrelia-specific Th1-response was further investigated in chronic, subacute and asymptomatic individuals by studying the expression of the Th1-marker IL-12Rβ2, on a protein and mRNA level. The cytokine secretion and Foxp3, a marker for regulatory T-cells, were also analyzed. Chronic patients had a lower IL-12Rβ2 expression on CD8+ T-cells and a lower number of Borrelia-specific IFN-γ secreting cells compared to asymptomatic individuals. Chronic patients also displayed a higher expression of Borrelia-specific Foxp3 than healthy controls. The conclusions for these tow studies were that a strong Th1-response early in the infection with a later switch to a Th2-response is beneficiary whereas a slow or weak Th1-response corresponds to a prolonged disease course. The influence of a previous infection with another pathogen, seen to suppress the immune response in animals, and the possible gender difference in immune response was also investigated. Patients with EM were screened for antibodies to Anaplasma phagocytophilum (Ap) as a sign of a previous exposure to these tick-borne bacteria. Blood lymphocytes from Ap seronegative, Ap seropositive and healthy controls were stimulated with Borrelia antigen and the secretion of IL-4, IL-5, IL-12, IL-13 and IFN-γ was detected by ELISPOT. Ap seropositive patients had a lower number of cells responding with IL-12 secretion compared to the other groups which might indicate an inhibited Th1-response. Reinfections with Lyme borreliosis was in a previous study, done by Bennet et al, found to be more frequent in postmenopausal women than in men. To investigate if there was an immunological explanation to the gender discrepancy, blood lymphocytes from individuals reinfected with Lyme borreliosis and individuals infected only once were stimulated with various antigens. The cytokine secretion was detected by ELISPOT, ELISA and Immulite. There were no differences between reinfected and single infected individuals. However, women, regardless of times infected, displayed a Th2-derived and anti-inflammatory spontaneous immune response compared to men. A previous infection with the bacteria Ap might possibly have a long term effect on the immune system and might be of disadvantage when mounting a Th1-response to a Borrelia infection. Also, the Th2-derived response displayed by postmenopausal women could indicate why more women than men get reinfected with Borrelia burgdorferi. / On the day of the public defence date of the doctoral thesis the status of article III was Accepted; the status of article IV was Submitted and the title was "Importance of induction and secretion of interferon-gamma for optimal resolution of human Lyme borreliosis: differencesbetween different outcomes of the infection".

Thesis (MSc (Production Animal Studies)--University of Pretoria, 2007. / Includes bibliographical references.

A anaplasmose bovina é uma das principais causas de perdas produtivas e mortes no Rio Grande do Sul em rebanhos bovinos. O Anaplasma marginale é o principal agente causador da enfermidade e provoca hipertermia, anemia, prostração, abortos e perdas produtivas nos animais acometidos. Tendo em vista o controle deste hemoparasita em uma propriedade leiteira localizada no município de Eldorado do Sul no Rio Grande do Sul, na qual a incidência da doença era alta, 473 animais foram imunizados com Anaplasma centrale na busca de desenvolvimento cruzado para Anaplasma marginale. No experimento foram verificadas que a incidência que normalmente era acima de 30% na propriedade passou para níveis inferires a 5%. No entanto, foram verificados abortos decorrentes da imunização, principalmente nos animais que possuíam menos de 90 dias de prenhes. Já o número de mortes globais na fazenda caiu consideravelmente tendo em vista que a principal causa de morte era a anaplasmose bovina. Dos animais inoculados com A. centrale em torno de 15% apresentaram sintomatologia clínica da enfermidade e precisaram ser tratados com oxitetraciclina no período entre 15 e 30 dias após a imunização. O custo com tratamento empregado na propriedade posterior à imunização caiu em torno de 85% o que provocou impacto significativo economicamente na propriedade. / Bovine anaplasmosis is a major cause of production losses and deaths in Rio Grande do Sul cattle herds. The Anaplasma is the main causative agent of cattle disease. It causes hyperthermia, anemia, prostration, abortions and reduces milk production in affected animals. In order to control this hemoparasite on a dairy farm located in the municipality of Eldorado do Sul in Rio Grande do Sul, where the disease incidence was high, 473 animals were immunized with Anaplasma centrale in the search for cross-protectiv immunity for Anaplasma marginale. The property anaplasmosis incidence, which usually was above 30%, became 5% after the immunization. However, abortions were observed resulting from innoculaition, especially in animals that had less than 90 days of pregnancy. The global number of deaths on the farm dropped considerably given that the main cause of death was the bovine anaplasmosis. 15% of animals inoculated with A. centrale showed clinical symptoms of the disease between 15 and 30 days after immunization and had to be treated with oxytetracycline . The amount of money spent with anaplasmosis treatment decay 85% after the immunization, which caused significant economic impact on the property.

La anaplasmosis es una enfermedad zoonótica emergente que es transmitida por la mordedura de garrapatas del género Ixodes. En nuestro país aún no existen reportes del agente Anaplasma phagocytophilum, más si hay evidencias de la garrapata vector, haciendo posible su presencia en el territorio. Por tanto, el objetivo del estudio fue detectar Anaplasma phagocytophilum en caballos de la provincia de Chiclayo (Departamento de Lambayeque, Perú) utilizando técnicas hematológicas. Para ello se colectaron 100 muestras de sangre de caballos procedentes de 4 haras en el año 2015, los cuales se encontraban aparentemente sanos y tenían antecedentes de exposición a garrapatas. El estudio hematológico evidenció que el 9% de caballos presentaron estructuras compatibles con cuerpos de inclusión o mórulas de Anaplasma phagocytophilum en frotices de sangre periférica teñidos con Wright y Naranja de acridina, a los que denóminamos “positivos”; además, este grupo de animales evidenciaron leves alteraciones en la serie leucocítica y trombocítica como lo reportaron otros estudios. A su vez, hubo 48% de animales sospechosos que presentaron trombocitopenia sin evidencia de cuerpos de inclusión o mórulas. Los hallazgos sugieren la exposición a A. phagocytophilum, constituyendo el primer reporte hematológico del agente en caballos en el Perú. / Tesis

A anaplasmose bovina é uma das principais causas de perdas produtivas e mortes no Rio Grande do Sul em rebanhos bovinos. O Anaplasma marginale é o principal agente causador da enfermidade e provoca hipertermia, anemia, prostração, abortos e perdas produtivas nos animais acometidos. Tendo em vista o controle deste hemoparasita em uma propriedade leiteira localizada no município de Eldorado do Sul no Rio Grande do Sul, na qual a incidência da doença era alta, 473 animais foram imunizados com Anaplasma centrale na busca de desenvolvimento cruzado para Anaplasma marginale. No experimento foram verificadas que a incidência que normalmente era acima de 30% na propriedade passou para níveis inferires a 5%. No entanto, foram verificados abortos decorrentes da imunização, principalmente nos animais que possuíam menos de 90 dias de prenhes. Já o número de mortes globais na fazenda caiu consideravelmente tendo em vista que a principal causa de morte era a anaplasmose bovina. Dos animais inoculados com A. centrale em torno de 15% apresentaram sintomatologia clínica da enfermidade e precisaram ser tratados com oxitetraciclina no período entre 15 e 30 dias após a imunização. O custo com tratamento empregado na propriedade posterior à imunização caiu em torno de 85% o que provocou impacto significativo economicamente na propriedade. / Bovine anaplasmosis is a major cause of production losses and deaths in Rio Grande do Sul cattle herds. The Anaplasma is the main causative agent of cattle disease. It causes hyperthermia, anemia, prostration, abortions and reduces milk production in affected animals. In order to control this hemoparasite on a dairy farm located in the municipality of Eldorado do Sul in Rio Grande do Sul, where the disease incidence was high, 473 animals were immunized with Anaplasma centrale in the search for cross-protectiv immunity for Anaplasma marginale. The property anaplasmosis incidence, which usually was above 30%, became 5% after the immunization. However, abortions were observed resulting from innoculaition, especially in animals that had less than 90 days of pregnancy. The global number of deaths on the farm dropped considerably given that the main cause of death was the bovine anaplasmosis. 15% of animals inoculated with A. centrale showed clinical symptoms of the disease between 15 and 30 days after immunization and had to be treated with oxytetracycline . The amount of money spent with anaplasmosis treatment decay 85% after the immunization, which caused significant economic impact on the property.

A anaplasmose bovina é uma das principais causas de perdas produtivas e mortes no Rio Grande do Sul em rebanhos bovinos. O Anaplasma marginale é o principal agente causador da enfermidade e provoca hipertermia, anemia, prostração, abortos e perdas produtivas nos animais acometidos. Tendo em vista o controle deste hemoparasita em uma propriedade leiteira localizada no município de Eldorado do Sul no Rio Grande do Sul, na qual a incidência da doença era alta, 473 animais foram imunizados com Anaplasma centrale na busca de desenvolvimento cruzado para Anaplasma marginale. No experimento foram verificadas que a incidência que normalmente era acima de 30% na propriedade passou para níveis inferires a 5%. No entanto, foram verificados abortos decorrentes da imunização, principalmente nos animais que possuíam menos de 90 dias de prenhes. Já o número de mortes globais na fazenda caiu consideravelmente tendo em vista que a principal causa de morte era a anaplasmose bovina. Dos animais inoculados com A. centrale em torno de 15% apresentaram sintomatologia clínica da enfermidade e precisaram ser tratados com oxitetraciclina no período entre 15 e 30 dias após a imunização. O custo com tratamento empregado na propriedade posterior à imunização caiu em torno de 85% o que provocou impacto significativo economicamente na propriedade. / Bovine anaplasmosis is a major cause of production losses and deaths in Rio Grande do Sul cattle herds. The Anaplasma is the main causative agent of cattle disease. It causes hyperthermia, anemia, prostration, abortions and reduces milk production in affected animals. In order to control this hemoparasite on a dairy farm located in the municipality of Eldorado do Sul in Rio Grande do Sul, where the disease incidence was high, 473 animals were immunized with Anaplasma centrale in the search for cross-protectiv immunity for Anaplasma marginale. The property anaplasmosis incidence, which usually was above 30%, became 5% after the immunization. However, abortions were observed resulting from innoculaition, especially in animals that had less than 90 days of pregnancy. The global number of deaths on the farm dropped considerably given that the main cause of death was the bovine anaplasmosis. 15% of animals inoculated with A. centrale showed clinical symptoms of the disease between 15 and 30 days after immunization and had to be treated with oxytetracycline . The amount of money spent with anaplasmosis treatment decay 85% after the immunization, which caused significant economic impact on the property.