Thèses sur le sujet « Babesiosi »

L'anaplasmosi, la babesiosi i la besnoitiosi es troben entre les malalties bovines poc conegudes i estudiades a Catalunya. A dia d'avui, més enllà de descripcions i comunicacions puntuals no disposem de cap estudi epidemiològic que ens permeti conèixer el seu estat actual. Amb el propòsit d'iniciar aquests estudis es planteja el present treball de Tesi Doctoral, centrat en l'estudi d'Anaplasma spp., Babesia bigemina i Besnoitia besnoiti. La prevalença dels dos hemoparàsits es va determinar a través de l’anàlisi de sèrums de diferents poblacions bovines. La metodologia va consistir en tècniques d’ELISA. L’estudi estadístic dels resultats i de diferents factors de risc associats a l’animal, el maneig i localització de l’explotació es va realitzar amb un model lineal generalitzat. Les seroprevalences observades van ser aproximadament d’un 40 % per a l’anaplasmosi i un 54,4 % per a la babesiosi causada per B. bigemina. L’edat i la cria en extensiu es van establir com a factors determinants. L'estudi de B. besnoiti consta a la vegada de dues parts. En la primera part, es van analitzar sèrums de bovins procedents de les zones nord i sud de Catalunya. mitjançant una tècnica d’ELISA L’estudi estadístic de les dades es va basar en un model lineal generalitzat. Els resultats de seroprevalença obtinguts van ser d'un 64 %. En la segona part de l'estudi de B. besnoiti es van obtenir mostres de globus oculars i de sang de 206 animals sacrificats en dos escorxadors de la comarca de la Garrotxa. Es van observar els quists escleroconjuntivals en un 3,9 % dels animals estudiats, mentre que la seroprevalença en els mateixos animals va ser d'un 29,6 %. Les seroprevalences obtingudes per a les hemoparasitosis i la besnoitiosi bovina mostren que s’haurien d’incloure en el diagnòstic diferencial de les malalties bovines a la nostra zona.
Anaplasmosis, babesiosis and besnoitiosis are bovine diseases with limited information in Catalonia. Nowadays, except for brief descriptions, no epidemiological studies are available to know the current status. The main objective of this Doctoral Thesis is to initiate these studies focused on Anaplasma spp., Babesia bigemina and Besnoitia besnoiti. Hemoparasites seroprevalence were established by the analysis of samples from different cattle population. The methodology consisted in ELISA analysis of serum samples. The statistical study of the results and different risk factors from animals, management and geographic location were obtained using a generalized linear model. The overall seroprevalence to Anaplasma spp. was closely to 40 % and to B. bigemina was 54.4 %. Age and extensive management were determined as determinant factors. B. besnoiti study was divided in two parts. In the first one, serum samples from north and south areas from Catalonia were analysed by an ELISA technique. Data analysis was based on a generalized linear model. The seroprevalence was found to be 64 %. In the second part of B. besnoiti study, eyeball and blood samples were collected from 206 slaughtered cattle in two abattoirs from Garrotxa region. 3,9 % of studied animals showed sclero-conjunctival cysts, whereas the seroprevalence was 29,6 % in the same animals. The high seroprevalences detected for both hemoparasites and bovine besnoitia suggested they all should be included in differential diagnosis in bovine diseases in our area.

Vector borne infections are now again a priority worldwide. Emergence or re-emergence of these diseases are usually associated with climatic change, but also other factor are involved, such as demographic, socio-economic and accidental factors. The results of 3 years epidemiolocal investigations about ruminants babesiosis and canine leishmaniosis in north-eastern Italy are presented. These two case studies show that these infections are a potential threat to Public Health. Outcomes of the research confirm the importance of a proper surveillance of vector-borne infections also in north-eastern Italy.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
A babesiose e a erliquiose canina são doenças comuns em cães, transmitidas pelo vetor Rhipicephalus sanguineus. Os sinais e sintomas dessas doenças não são específicos. A detecção dos parasitos em esfregaços sanguíneos nem sempre é possível e a detecção sorológica pode indicar apenas que o animal teve contato com o agente, mas não necessariamente a doença ativa. Na Colômbia existem poucas pesquisas sobre a epidemiologia e diagnóstico de erliquiose e babesiose em cães, as poucas existentes se basearam na detecção dos hemoparasitas em esfregaços sanguíneos, ou de anticorpos específicos. Sendo assim propos-se este estudo com o objetivo de pesquisar a erliquiose e a babesiose em esfregaços sangüíneos associados a provas sorológicas e moleculares em cães da região centro-oriental de Colômbia. Amostras de sangue e soro de 91 cães, provenientes das cidades de Bogotá (n=21), Villavicencio (n=31) e Bucaramanga (n=39) foram submetidas a pesquisa parasitológica, detecção de anticorpos anti-B. canis e anti-E. canis e detecção molecular de Babesia canis e Ehrlichia canis. Os resultados mostraram que dentre as 91 amostras testadas, 4% e 5% delas apresentaram E. canis e B. canis, respectivamente nas pesquisas parasitológicas dos esfregaços sanguíneos. Nas mesmas amostras, 82% e 51% delas apresentaram anticorpos anti-E. canis e anti-B. canis, respectivamente e, em 40% e 5%, foram detectados os DNA de E. canis e de B. canis, respectivamente. Os amplicons foram confirmados por seqüenciamento e o DNA de E. canis e de B. canis obtidos mostraram similaridade genética de 98% a 99% com amostras de E. canis e de B. canis isoladas em outros países, as árvores filogenéticas dos agentes pesquisados foram
Canine babesiosis and ehrlichiosis are common diseases in dogs, and are transmitted by the vector Rhipicephalus sanguineus. The clinical signs of these diseases are not specific. Detection of parasites in blood smears is not always possible and the serological detection could only indicate that the animal has had contact with the agent and not necessarily that the disease is being developed. In Colombia, there are few reports on canine babesiosis and ehrlichiosis´s epidemiology and diagnosis, being them based only on the detection of haemoparasites in blood smears and detection of antibodies. The purpose of the present study was to investigate the presence of Ehrlichia sp. and Babesia sp. in blood smears, associated with serological and molecular detection of these parasites in dog blood samples from the middle east region of Colombia. Blood and sera samples from 91 dogs from Bogotá (n=21), Villavicencio (n=31) and Bucaramanga cities (n=39) were submitted to parasitological, serological and molecular detection of B. canis and E. canis. Among the 91 samples tested, 4% and 5% presented E. canis morulae and B. canis piroplasms in blood smears, respectively. Moreover, 82% and 51% of samples presented antibodies anti-E. canis and anti-B. canis, respectively. E. canis and B. canis DNA were detected in 40% and 5% of samples, respectively. The amplicons were confirmed by sequencing and the found DNA from E. canis and B. canis showed a genetic identity of 98% to 99% among samples from E. canis and B. canis isolated from other countries. Finally, phyllogenetic trees were constructed from both agents

O relatório de estágio de Mestrado Integrado em Medicina Veterinária, aqui apresentado, encontra-se dividido em duas secções. A primeira corresponde à descrição das atividades médico-veterinárias desenvolvidas ao longo do estágio curricular e a segunda secção corresponde à revisão bibliográfica de babesiose canina. O estágio foi realizado no Hospital Animal do Sul, situado em Faro e abrangeu a área de medicina interna e cirurgia de animais de companhia. A revisão bibliográfica é seguida do relato e discussão de um caso clínico de babesiose não complicada, acompanhado durante o período de estágio. A babesiose canina faz parte das doenças transmitidas por vetores com grande importância mundial e, apesar de ainda não estarem descritos casos humanos provocados por espécies de Babesia responsáveis por doença em cães, existem outros fatores que devem ser tidos em conta na análise da babesiose canina como uma futura potencial zoonose; Abstract Canine Babesiosis This report of Integrated Masters Degree in Veterinary Medicine, here presented, is divided into two sections. The first one corresponds to the description of medicalveterinary activities developed throughout the internship and the second one corresponds to a literature review on canine babesiosis. The internship was perfomed in Hospital Animal do Sul, in Faro and has as main areas of interest internal medicine and surgery of domestic animals. The literature review is followed by a presentation and discussion of a case-report of a uncomplicated babesiosis case, followed during the internship period. Canine babesiosis is one of vector-borne diseases of worldwide importance and, although human cases caused by species of Babesia responsible for disease in dogs, haven’t been reported, there are other factors that should be considered in the analysis of canine babesiosis as a future potential zoonosis.

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This study was conducted with the purpose to evaluate the clinical, laboratory and epidemiological aspects of naturally acquired Babesia equi infection in 178 horses handled in a semi-intensive system, property of the Academia Militar das Agulhas Negras Brazilian Army, situated in the city of Resende, in the Rio de Janeiro State. Babesia equi is a species of hemoparasite of elevated prevalence among equids in tropical regions, being hard to control, causing important economic losses because of the long periods of convalescence, decline of performance in the affected animals, expenses with medication and workers, besides the restrictions in transit of equines, either for hippic competitions or international commerce. In the Indirect Immunofluorescence Reaction, a prevalence of 85,96% (n=153) was verified, therefore the area can be considered endemic for Babesia equi and in enzootic stability. There was no significant difference in the age or sex of the seropositive animals. Laboratory parameters like packed cell volume, red blood cell count, platelet count, global and differential white blood cell counts, dosages of seric creatinine, albumin and total plasmatic proteins, all of which led to the conclusion that in the assymptomatic carrier state there is no hematologic or seric biochemical variations that could be attributed to the protozoal Babesia equi.
Este estudo foi conduzido com o intuito de avaliar achados cl?nicos, laboratoriais e epidemiol?gicos na infec??o natural de Babesia equi em 178 eq?inos mantidos em sistema semiintensivo de cria??o, de propriedade da Academia Militar das Agulhas Negras Ex?rcito Brasileiro, situada no munic?pio de Resende, Estado do Rio de Janeiro. Babesia equi ? uma esp?cie de hemoparasita de elevada preval?ncia entre eq??deos de ?reas tropicais, sendo de dif?cil controle, causando perdas econ?micas importantes, pelos longos per?odos de convalescen?a, queda de desempenho nos animais acometidos, gastos com medicamentos e m?o de obra, al?m de impor restri??es no tr?nsito de eq?inos, seja para competi??es h?picas ou com?rcio internacional. Na Rea??o de Imunofluoresc?ncia Indireta, foi verificada uma preval?ncia de 85,96% (n=153), podendo ser a ?rea considerada end?mica para Babesia equi e em estabilidade enzo?tica, n?o sendo observadas diferen?as significativas entre sexo e idade dos animais. Par?metros laboratoriais como volume globular, hematimetria, plaquetometria, leucometrias global e espec?fica, dosagens de creatinina s?rica, albumina e prote?nas plasm?ticas totais foram conduzidos, podendo-se concluir que no estado de portador assintom?tico n?o h? varia??o hematol?gica ou de bioqu?mica s?rica que possa ser creditada ao protozo?rio Babesia equi.

Dissertação de Mestrado Integrado em Medicina Veterinária
As doenças transmitidas por vectores são causadas por parasitas, bactérias ou vírus e são veiculadas pela picada de artrópodes (principalmente ixodídeos e mosquitos). Estas doenças afectam cães a nível mundial e a maioria delas têm potencial zoonótico. Nestas estão incluídas as doenças sob estudo: Erliquiose, Babesiose, Riquetsiose, Anaplasmose e Leishmaniose. A patogénese das doenças sob estudo, baseia-se fundamentalmente na resposta humoral exagerada e não protectora desenvolvida pelo hospedeiro, que provoca sintomas semelhantes aos de uma doença auto-imune. Isto é, ao contrário do que se podia pensar, não é o parasita em si que é responsável por todos os sintomas da doença, mas sim a resposta imunológica do hospedeiro à sua presença. No que respeita ao diagnóstico, os médicos veterinários podem ter a sua tarefa complicada devido à ausência de sintomas específicos e à presença de co-infecções (infecção por mais do que um parasita). A compreensão por parte do clínico, da patogenia e imunologia das doenças transmitidas por vectores, é uma ferramenta essencial para o diagnóstico rápido e preciso. Sob o ponto de vista zoonótico, se por um lado os canídeos actuam como sentinelas de doenças, por outro, são reservatórios de parasitas e importantes hospedeiros de transporte de vectores. Devido a isto um dos pontos fulcrais da prevenção destas hemoparasitoses é o controlo de vectores. Neste estudo foi realizado o rastreio de cinco doenças transmitidas por vectores, causadas pelos agentes: Ehrlichia canis, Anaplasma phagocytophilum, Rickettsia conorii, Babesia canis e Leishmania infantum. Este estudo incidiu sobre uma população de um canil de Setúbal, no qual foram testados 80 cães (29 machos e 51 fêmeas) escolhidos aleatoriamente. Os testes utilizados foram testes comerciais de imunofluorescência indirecta (IFI). Dos animais testados 58,75% dos animais encontravam-se infectados (20% co-infectados e 38,75% mono-infectados). As prevalências de anticorpos contra os 5 agentes foram: Rickettsia conorii (23,75%), Babesia canis (20%), Ehrlichia canis (16,25%), Leishmania infantum (16,25%) e Anaplasma phagocytophilum (12,5%).
ABSTRACT - Vector-borne diseases in kennel dogs, a study - Vector-borne diseases are caused by parasites, virus and bacteria, and transmitted through the bite of arthropods. These diseases have a great zoonotic potential and affect dogs worldwide. The five diseases studied (Ehrlichiosis, Babesiosis, Rickettsiosis, Anaplasmosis and Leishmaniasis) are included in this category of illness. The pathogenesis, it‘s based mainly on the humoral response developed by the host, that being exaggerated and not protective, causes symptoms similar to an autoimmune disease. Meaning, on the contrary to what might be thought, is not the parasite itself that is responsible for all the symptoms, but the host‘s immune response to their presence. Concerning the diagnosis, veterinarians can have their task complicated by the absence of specific symptoms and the presence of co-infection (infection for more than a parasite).The understanding of the clinical, the pathogenesis and immunology of vector-borne diseases, is an essential tool for rapid and accurate diagnosis. From the zoonotic point of view, dogs act as sentinels of disease, and are important reservoirs of parasites and carry vectors. Due to this, one of the key points for prevention of vector-borne diseases in animals and humans is vector control. In this study was performed the screening of five vector-borne diseases caused by the agents: Ehrlichia canis, Anaplasma phagocytophilum, Rickettsia conorii, Babesia canis and Leishmania infantum. This study focused on a population of a kennel of Setúbal, in which 80 dogs (29 males and 51 females), randomly chosen, were tested. The tests used were indirect fluorescence antibody (IFA) commercial. 58.75% of the animals tested animals were infected (20% co-infected and 38.75% single infections). The point prevalence for different pathogens was: Rickettsia conorii (23.75%), Babesia canis (20%), Ehrlichia canis (16.25%), Leishmania infantum (16.25%) and Anaplasma phagocytophilum (12.5%).

Dissertação de Mestrado Integrado em Medicina Veterinária
Este estudo foi realizado com os seguintes objetivos: a) compreender a ocorrência de Babesiose Canina (BC) e Leishmaniose Canina (LCan) nos cães nos concelhos de Lamego, Tarouca e Peso da Régua; b) avaliar o risco de contacto dos cães com os vetores; c) analisar as medidas profiláticas instituídas nos animais amostrados. Para obtermos informação sobre as características da vida dos animais foi realizado um inquérito aos proprietários dos animais. Ao todo foram preenchidos 60 inquéritos (1 por animal). A amostra de 60 animais foi constituída por 32 cães diagnosticados com BC e 28 cães que foram submetidos a diagnóstico, rastreio ou avaliação do controlo de LCan. Relativamente aos animais amostrados para BC, verificámos que 100% dos animais foram positivos a Babesia spp, enquanto 60,7% dos cães amostrados para LCan foram positivos para Leishmania sp. O diagnóstico de BC foi realizado através da visualização de formas intraeritrocitárias de Babesia spp. no esfregaço sanguíneo dos 32 animais. Os testes utilizados para diagnóstico, rastreio e controlo de LCan variaram entre o método de ELISA e um teste imunocromatográfico. O teste de ELISA foi utilizado em 71,4% da população amostrada ao passo que o teste imunocromatográfico foi empregue em 28,6% dos cães. Relativamente às duas doenças estudadas verificámos que 56,3% dos animais amostrados para BC eram provenientes do concelho de Lamego, ao passo que 42,9% dos animais estudados para LCan pertenciam ao concelho do Peso da Régua. O estudo decorreu entre o dia 5 de setembro de 2016 e 28 de fevereiro de 2017, e verificámos que novembro de 2016 foi o mês em que se realizaram mais diagnósticos de BC, enquanto fevereiro de 2017 foi o mês em que se realizaram mais testes para diagnóstico, rastreio ou controlo de LCan. Alguns dos factores estudados para avaliar o risco de contacto com os vetores das respetivas doenças foram: a aptidão do animal, local onde dormem, convivência com cães diagnosticados com BC e LCan, acesso ao exterior das habitações, entre outros. Para análise da profilaxia dos animais comparámos a frequência utilizada com a recomendada pelo fabricante dos respetivos produtos. Assim dos 32 animais amostrados para Babesiose Canina verificámos que nenhum dos animais estava corretamente desparasitado, ao passo que apenas 39,3% dos animais testados para LCan apresentavam proteção correta para ectoparasitas.
ABSTRACT - Babesiosis and Leishmaniaosis study in domestic canids from the counties of Lamego, Tarouca and Peso da Régua, Portugal. - This study was accomplished based on the following goals: a) understand the occurrence of Canine Babesiosis (CB) and Canine Leishmaniasis (CanL) in domestic canids from the counties of Lamego, Tarouca and Peso da Régua; b) assess the risk of contact with the vectors; c) to analyse the prophylactic measures established in the sampled animals. In order to obtain information about the characteristics of the animals’ life, a survey was performed with the dog owners, resulting in 60 fulfilled inquiries, one per animal. This sample was constituted by 32 canids diagnosed with CB and 28 canids subbmited to a CanL diagnosis, survey and control assessment. Concerning the animals sampled for CB, we verified that 100% of the animals were positive to Babesia spp, while 60.7% of the dogs sampled to CanL were positive for Leishmania sp.. The CB diagnosis was performed using visualisation of intra-erythrocyte forms of Babesia spp. in the blood smear of the 32 animals. The tests used for the CanL diagnosis, survey and control assessment varied between the ELISA’s method and an immunochromatographic test. The first was used in 71,4% of the samples population and the second in 28,6% of the canids. Regarding the two studied diseases, we found that 56,3% of the CB samples animals originated from the county of Lamego whilst 42,9% of the CanL sampled animals belonged to the Peso da Régua county. The study was conducted between 5th September 2016 and 28th February 2017, and we verified that November 2016 was the month with more CB diagnosis, while February 2017 was the month in which more CanL diagnosis tests, survey and control assessment were performed. Some of the factors studied to assess the risk of contact with the vectors concerning these diseases were: the aptitude of the animal, the place where they slept, the coexistence with canids diagnosed with CB and CanL, the access to the outside of dwellings, among others. After analysing the animals prophylaxis, we compared the frequency used with the one recommended by the manufacturers of the respective products. Thus, among the 32 canine Babesiosis sampled animals we verified that none of those animals were properly dewormed, whilst only 39,3% of the CanL tested animals showed a correct protection regarding ectoparasites.
N/A

Orientadora: Mirela Tinucci Costa
Banca: Gervásio Henrique Bechara
Banca: Paulo Cesar Ciarlini
Resumo: A babesiose e a erliquiose canina são doenças comuns em cães, transmitidas pelo vetor Rhipicephalus sanguineus. Os sinais e sintomas dessas doenças não são específicos. A detecção dos parasitos em esfregaços sanguíneos nem sempre é possível e a detecção sorológica pode indicar apenas que o animal teve contato com o agente, mas não necessariamente a doença ativa. Na Colômbia existem poucas pesquisas sobre a epidemiologia e diagnóstico de erliquiose e babesiose em cães, as poucas existentes se basearam na detecção dos hemoparasitas em esfregaços sanguíneos, ou de anticorpos específicos. Sendo assim propos-se este estudo com o objetivo de pesquisar a erliquiose e a babesiose em esfregaços sangüíneos associados a provas sorológicas e moleculares em cães da região centro-oriental de Colômbia. Amostras de sangue e soro de 91 cães, provenientes das cidades de Bogotá (n=21), Villavicencio (n=31) e Bucaramanga (n=39) foram submetidas a pesquisa parasitológica, detecção de anticorpos anti-B. canis e anti-E. canis e detecção molecular de Babesia canis e Ehrlichia canis. Os resultados mostraram que dentre as 91 amostras testadas, 4% e 5% delas apresentaram E. canis e B. canis, respectivamente nas pesquisas parasitológicas dos esfregaços sanguíneos. Nas mesmas amostras, 82% e 51% delas apresentaram anticorpos anti-E. canis e anti-B. canis, respectivamente e, em 40% e 5%, foram detectados os DNA de E. canis e de B. canis, respectivamente. Os amplicons foram confirmados por seqüenciamento e o DNA de E. canis e de B. canis obtidos mostraram similaridade genética de 98% a 99% com amostras de E. canis e de B. canis isoladas em outros países, as árvores filogenéticas dos agentes pesquisados foram
Abstract: Canine babesiosis and ehrlichiosis are common diseases in dogs, and are transmitted by the vector Rhipicephalus sanguineus. The clinical signs of these diseases are not specific. Detection of parasites in blood smears is not always possible and the serological detection could only indicate that the animal has had contact with the agent and not necessarily that the disease is being developed. In Colombia, there are few reports on canine babesiosis and ehrlichiosis's epidemiology and diagnosis, being them based only on the detection of haemoparasites in blood smears and detection of antibodies. The purpose of the present study was to investigate the presence of Ehrlichia sp. and Babesia sp. in blood smears, associated with serological and molecular detection of these parasites in dog blood samples from the middle east region of Colombia. Blood and sera samples from 91 dogs from Bogotá (n=21), Villavicencio (n=31) and Bucaramanga cities (n=39) were submitted to parasitological, serological and molecular detection of B. canis and E. canis. Among the 91 samples tested, 4% and 5% presented E. canis morulae and B. canis piroplasms in blood smears, respectively. Moreover, 82% and 51% of samples presented antibodies anti-E. canis and anti-B. canis, respectively. E. canis and B. canis DNA were detected in 40% and 5% of samples, respectively. The amplicons were confirmed by sequencing and the found DNA from E. canis and B. canis showed a genetic identity of 98% to 99% among samples from E. canis and B. canis isolated from other countries. Finally, phyllogenetic trees were constructed from both agents
Mestre

Orientador: Teresa Cristina Goulart de Oliveira Sequeira
Banca: Claudio Roberto Madruga
Banca: Márcia Cristina Sena de Oliveira
Banca: Lúcia Helena O'Dwyer
Banca: Agueri Kohayagawa
Resumo: A dinâmica das infecções naturais por Babesia bovis, Babesia bigemina e Anaplasma marginale, foi investigada ao longo do primeiro ano de vida de bezerros da raça Nelore criados em sistema extensivo no Estado de São Paulo, com o propósito de se avaliar a influência da criação exclusiva e extensiva de bovinos da raça Nelore sobre a epidemiologia desses hemoparasitas. Amostras de sangue de 17 animais P.O. foram colhidas a cada 15 dias do nascimento até os quatro meses, e então, a cada 30 dias até um ano. Amostras de sangue colhidas da ponta da orelha foram utilizadas para a confecção de esfregaços sanguíneos, e amostras de sangue da veia jugular foram utilizadas para a determinação da contagem total de eritrócitos (RBC), concentração de hemoglobina (HB), volume globular (VG), contagem total de leucócitos (CTL) e extração de DNA. As teleóginas colhidas foram incubadas em estufa BOD e posteriormente submetidas ao processo de extração de DNA. A amplificação do DNA dos hemoparasitas foi feita pelas técnicas de PCR e nPCR, utilizando-se respectivamente os primers BoF e BoR; BoFN e BoRN para Babesia bovis, BiIA e BilB; BilAN e BilBN para B. bigemina e Am9 e Am10; Am11 e Am12 para A. marginale. Ao longo de um ano de experimento não foram observados casos clínicos de tristeza parasitária, e os quatro animais em que parâmetros hematológicos indicativos de anemia foram detectados concomitantemente com uma ou mais espécies de hemoparasitas se recuperaram sem que houvesse necessidade de tratamento. Babesia bovis não foi detectada nos esfregaços sangüíneos corados por Giemsa, porém merozoítas de B. bigemina foram encontrados em 5 amostras de sangue provenientes de 4 animais. Corpúsculos intraeritrocitários de A. marginale foram detectados em 29 amostras oriundas de 14 animais, nas quais a mais elevada taxa de parasitemia foi de 26%... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The dynamics of natural Babesia bovis, Babesia bigemina and Anaplasma marginale infection was investigated along the first year of Nelore calves, raised on extensive system in São Paulo State. The purpose was to evaluate the influence of the exclusive and extensive breeding of Nelore bovine on the epidemiology of those hemoparasites. Blood samples of 17 purebred animals were harvested every 15 days beginning on the first day of birth till four months old, and then, every 30 days till one year old. Blood samples from the ear tip were used to made blood smears, and blood samples were taken from the jugular vein to determine the red blood cells total count (RBC), hemoglobin concentration (HB), haematocrite (VG), leukocytes total count (LTC) and DNA extraction. The engorged Boophilus microplus were incubated and submitted to the DNA extraction process. The hemoparasite DNA amplification was made by PCR and nPCR respectively, using the primers BoF and BoR; BoFN and BoRN for Babesia bovis, BiIA and BilB; BilAN and BilBN for B. bigemina and Am9 and Am10; Am11 and Am12 for A. marginale. Clinical cases of parasitic sickness were not observed along one year of experiment. The four animals that were observed with indication of anemia hematological parameters, detected at the same time with one or more hemoparasites species, recovered without treatment. Babesia bovis was not detected in the blood smears. However, B. bigemina merozoites were found in five blood samples coming from four animals. A. marginale was detected in 29 blood samples of 14 animals, in which the highest parasitemia level was 26%. The reactions of PCR and nPCR revealed the presence of B. bovis DNA in seven blood samples (four by PCR) from three animals, one of them born in less than 24 hours... (Complete abstract, access undermentioned electronic address)
Doutor

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil.
This study aimed to evaluate the influence of babesiosis in embryo recovery rates, early pregnancy and embryonic loss. The project was carried out in a Central Commercial Embryo Transfer, located in the city of Itaguai, and two farms in the county of Serop?dica-RJ, using animals from the same breed (Mangalarga Marchador). The 13 donors and 40 recipients used in this experiment were positive for Theileria equi confirmed by nested-PCR method . Two embryo collections were performed in donor mares in two consecutive estrous cycles (GID), in sequence these same animals were treated with imidocarb dipropionate (1.2 mg / kg IM.) in order to conduct two more embryo collections in two estrous cycles (GIId). The embryo recipients were divided into two groups of 20 animals each, where one was the control group (IRG), and the other group was treated (GIIr) with 1.2 mg / kg IM imidocarb dipropionate, with the aim of evaluating pregnancy rate at 15, 30, 45 and 60 days. The embryo donors mares had normal erythrocyte before and after treatment with imidocarb dipropionate. ConcerningWBC, the embryo donors showed a decrease in the count of total leukocytes and neutrophils after treatment and a slight increase of lymphocytes and monocytes after treatment. The embryo recipients showed normal erythrocyte and leukocyte counts before and after the treatment with imidocarb dipropionate. After 52 embryo collections performed, embryo recovery rate was 53.84% (14/26) and 65.38% (17/26) ( p> 0,05) for GID and GIId respectively. The pregnancy rate was 70% (14/20) ( p> 0,05) at 15, 30, 45 and 60 days in the Gir and the GIIr was 85% (17/20) ( p> 0,05) at 15 days, 80% (16/20) ( p> 0,05) at 30 , 45 and 60 days. Treatment with imidocarb dipropionate did not improve significantly the reproductive efficiency in an ET program.
Este estudo teve por objetivo avaliar a influ?ncia da babesiose nas taxas de recupera??o embrion?ria, gesta??o e perda embrion?ria precoce. O projeto foi realizado em uma Central Comercial de Transfer?ncia de Embri?o, situado no Munic?pio de Itagua?, e em dois haras no munic?pio de Serop?dica- RJ. Foram utilizadas 13 doadoras e 40 receptoras de embri?o da ra?a Mangalarga Marchador, positivas para Theileria equi atrav?s do m?todo de nested-PCR. Nas ?guas doadoras foram realizados duas coletas de embri?es em dois ciclo estrais consecutivos (GId), em sequ?ncia, esses mesmos animais foram tratados com dipropionato de imidocarb (1,2 mg/kg IM.) para realiza??o de mais duas coletas de embri?es em dois ciclos estrais (GIId). As receptoras de embri?o foram divididas em dois grupos de 20 animais cada, onde um grupo foi o controle (GIr) e, o outro grupo, foi tratado (GIIr) com 1,2 mg/ Kg IM de dipropionato de imidocarb, com intuito de avaliar a taxa de gesta??o aos 15, 30, 45 e 60 dias. As ?guas doadoras apresentaram eritrograma normal antes e ap?s o tratamento com dipropionato de imidocarb, e em rela??o ao leucograma, apresentaram diminui??o na contagem de leuc?citos e neutr?filos totais e discreto aumento de linf?citos e mon?citos somente ap?s o tratamento. As receptoras apresentaram eritrograma e leucograma normal antes e ap?s o tratamento com dipropionato de imidocarb. Ap?s a realiza??o de 52 coletas de embri?o, a taxa de recupera??o embrion?ria foi 53,84% (14/26) e 65,38% (17/26) (p> 0,05) para GId e GIId respectivamente. A taxa de gesta??o foi de 70% (14/20) (p> 0,05) aos 15, 30, 45 e 60 dias no grupo GIr e para o GIIr foi 85% (17/20) (p> 0,05) aos 15 dias, 80% (16/20) (p>0,05) aos 30, 45 e 60 dias. O tratamento com dipropionato de imidocarb na forma como foi realizado n?o promoveu melhora significativa na efici?ncia reprodutiva em um programa de TE.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
A dinâmica das infecções naturais por Babesia bovis, Babesia bigemina e Anaplasma marginale, foi investigada ao longo do primeiro ano de vida de bezerros da raça Nelore criados em sistema extensivo no Estado de São Paulo, com o propósito de se avaliar a influência da criação exclusiva e extensiva de bovinos da raça Nelore sobre a epidemiologia desses hemoparasitas. Amostras de sangue de 17 animais P.O. foram colhidas a cada 15 dias do nascimento até os quatro meses, e então, a cada 30 dias até um ano. Amostras de sangue colhidas da ponta da orelha foram utilizadas para a confecção de esfregaços sanguíneos, e amostras de sangue da veia jugular foram utilizadas para a determinação da contagem total de eritrócitos (RBC), concentração de hemoglobina (HB), volume globular (VG), contagem total de leucócitos (CTL) e extração de DNA. As teleóginas colhidas foram incubadas em estufa BOD e posteriormente submetidas ao processo de extração de DNA. A amplificação do DNA dos hemoparasitas foi feita pelas técnicas de PCR e nPCR, utilizando-se respectivamente os primers BoF e BoR; BoFN e BoRN para Babesia bovis, BiIA e BilB; BilAN e BilBN para B. bigemina e Am9 e Am10; Am11 e Am12 para A. marginale. Ao longo de um ano de experimento não foram observados casos clínicos de tristeza parasitária, e os quatro animais em que parâmetros hematológicos indicativos de anemia foram detectados concomitantemente com uma ou mais espécies de hemoparasitas se recuperaram sem que houvesse necessidade de tratamento. Babesia bovis não foi detectada nos esfregaços sangüíneos corados por Giemsa, porém merozoítas de B. bigemina foram encontrados em 5 amostras de sangue provenientes de 4 animais. Corpúsculos intraeritrocitários de A. marginale foram detectados em 29 amostras oriundas de 14 animais, nas quais a mais elevada taxa de parasitemia foi de 26%...
The dynamics of natural Babesia bovis, Babesia bigemina and Anaplasma marginale infection was investigated along the first year of Nelore calves, raised on extensive system in São Paulo State. The purpose was to evaluate the influence of the exclusive and extensive breeding of Nelore bovine on the epidemiology of those hemoparasites. Blood samples of 17 purebred animals were harvested every 15 days beginning on the first day of birth till four months old, and then, every 30 days till one year old. Blood samples from the ear tip were used to made blood smears, and blood samples were taken from the jugular vein to determine the red blood cells total count (RBC), hemoglobin concentration (HB), haematocrite (VG), leukocytes total count (LTC) and DNA extraction. The engorged Boophilus microplus were incubated and submitted to the DNA extraction process. The hemoparasite DNA amplification was made by PCR and nPCR respectively, using the primers BoF and BoR; BoFN and BoRN for Babesia bovis, BiIA and BilB; BilAN and BilBN for B. bigemina and Am9 and Am10; Am11 and Am12 for A. marginale. Clinical cases of parasitic sickness were not observed along one year of experiment. The four animals that were observed with indication of anemia hematological parameters, detected at the same time with one or more hemoparasites species, recovered without treatment. Babesia bovis was not detected in the blood smears. However, B. bigemina merozoites were found in five blood samples coming from four animals. A. marginale was detected in 29 blood samples of 14 animals, in which the highest parasitemia level was 26%. The reactions of PCR and nPCR revealed the presence of B. bovis DNA in seven blood samples (four by PCR) from three animals, one of them born in less than 24 hours... (Complete abstract, access undermentioned electronic address)

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico
This study was conducted with the objective to contribute with the agreement of the relation calves x ticks x hemoparasites in the sector of milk cows of the Farm of the Institute of Zootecnia (FAIZ) of the Universidade Federal Rural do Rio de Janeiro (UFRRJ). Seventeen female calves with age between 15 days old and 14 months old, between july of 2006 and june of 2007. These animals were subdivided in three ages bands: up to 2 months, between 3- 6 months and above of 7 months, in accordance with the handling of the property. Was realized ticks`s counting, collection of blood and hematological examination of all the animals in interval of 14 days. The exams were carrying through in laboratories of Parasites Diseases, Clinical Pathology of UFRRJ and the Serological of Embrapa Beef Cattle. Throughout 12 months, it can be verified the constant presence of larvaes, nymphs and females of tick Rhipicephalus (Boophilus) microplus. The frequency of positive animals for the indirect enzyme-linked immunosorbent assay (ELISA), for the agents of the Tick-borne Disease (Babesia bigemina, B. bovis and Anaplasma marginale) and Borrelia sp., was verified that in all ages bands exist positive serological animal. The frequency and antibody levels, as much for B. bigemina as for B. bovis, evaluated through the indirect ELISA, had been high. This fact associated with the absence of infection symptoms suggests a situation of immunization of the animals and an area of enzootically stable. None trend of seasonal distribution of infections for B. bigemina, B. bovis, A. marginale and Borrelia sp. was observed.
Este estudo foi conduzido com o objetivo de contribuir para o entendimento das rela??es bezerros x carrapatos x hemoparasitos no setor de bovinocultura de leite da Fazenda do Instituto de Zootecnia (FAIZ) da Universidade Federal Rural do Rio de Janeiro (UFRRJ). Foram utilizadas 17 bezerras com idade entre 15 dias e 14 meses, entre julho de 2006 a junho de 2007. Estes animais foram subdivididos em tr?s faixas et?rias: at? 2 meses, de 3-6 meses e acima de 7 meses, de acordo com o manejo zoot?cnico da propriedade. Procedeu-se contagem de carrapatos, coleta de sangue e exames hematol?gicos de todos animais em intervalo de 14 dias. Os exames foram realizados nos Laborat?rios de Doen?as Parasit?rias, de Patologia Cl?nica da UFRRJ e de Sorologia da Embrapa Gado de Corte. Ao longo de 12 meses, podese verificar a presen?a constante de larvas, ninfas e f?meas de carrapatos Rhipicephalus (Boophilus) microplus. Em rela??o a freq??ncia de positividade pelo ensaio de imunoadsors?o enzim?tica (ELISA) indireto, para os agentes da Tristeza Parasit?ria Bovina (Babesia bigemina, B. bovis e Anaplasma marginale) e Borrelia sp., verificou-se que em todas as faixas et?rias haviam animais sorologicamente positivos. A freq??ncia e os n?veis de anticorpos, tanto para B. bigemina como para B. bovis, avaliados atrav?s do ELISA indireto, foram altos. Este fato associado ? aus?ncia de sintomas de infec??o sugere uma situa??o de pr?-imuniza??o dos animais e uma ?rea de estabilidade enzo?tica. N?o foi observada qualquer tend?ncia de distribui??o sazonal de infec??es por B. bigemina, B. bovis, Anaplasma marginale e Borrelia sp.

O dipropionato de imidocar (D. I.) é um medicamento empregado tanto no tratamento como na profilaxia da babesiose. O objetivo com esse trabalho foi avaliar os possíveis efeitos embriotóxicos da administração do D. I. a ratas durante o período de organogenese. Para tanto, foram utilizadas sessenta ratas divididas em três grupos dois experimentais e um controle, que receberam do sexto ao décimo quinto dia de gestação por via subcutânea, duas doses terapêuticas do D. I. a de 1,7 mg/kg e a de 2,5 mg/kg e o grupo controle NaCl a 0,9%. Ambas doses não alteraram o peso e o ganho de peso materno, durante o período de administração, indicando ausência de toxicidade materna. Na dose de 2,5 mg/kg do D. I. promoveu aumento da média de peso dos filhotes por ninhada e diminuição da média de peso da placenta individual e por ninhada. Em relação à avaliação óssea e visceral não foram detectadas diferenças entre os grupos experimentais e controle. Na dosse de 2,5 mg/kg do D. I. notou-se também maior maturidade da prole indicada por uma maior ossificação total e de número de esternébrios
The imidocarb dipropionate (I. D.) drug is widely used in the treatment as well in the profilaxy of the babesiosis. The treatment of this study was to investigative a possible embriotoxic effects of I. D. administered during organogenic period in rats. Sixty rats were used and divided into three groups, two experimental groups and one control group. Animals of the experimental groups received (sc), 1,7 mg/kg or 2,5 mg/kg of the I. D. from 6º to 15º days of pregnancy. The control group received only the saline solution. Results showed that both I. D. doses were unable to induce changes in the maternal weight and maternal weight gain during the treatment period, showing no maternal toxicity. At the I. D. 2,5 mg/kg dose group an increase on the fetal weight was observed by brood and a decrease on the placental weight was observed either individually or by brood. The skeletal and visceral evaluation were not modified by the prenatal exposure. However, at the I. D. 2,5 mg/kg dose and improvement in the offspring maturity was observed by an increase of the skeletal total ossification and by the number of esternebrios

Canine babesiosis typically causes a haemolytic anemia and results in hypoxia and sepsis, which can eventually result in multiple organ dysfunction. Human patients with severe injury or disease such as shock, sepsis and malaria often have persistent hyperlactataemia, and there is a correlation between blood lactate and survival rate. There are various similarities between human malaria and canine babesiosis, eg. anaemia, renal failure, cerebral forms, coagulopathy, hepatopathy, pulmonary oedema, and shock. In severe malaria, lactate levels in blood rise in direct proportion to the severity of the disease. Venous lactate concentrations measured at 4 hours after admission appears to be the best prognostic indicator in severe malaria. In dogs blood lactate has been shown to be of prognostic value in patients with gastric dilatationvolvulus and in dogs admitted to intensive care units. Blood lactate has also been shown to be of prognostic value in equine colic. Blood lactate was determined in ninety dogs with naturally occurring canine babesiosis. Forty-five dogs (50%) presented with hyperlactataemia (blood lactate > 2.5mmol/L) and 20 (22.2%) with hypoglycaemia (blood glucose < 3.3 mmol/L). Measurements significantly associated with mortality were hypoglycaemia on admission, blood lactate > 5mmol/L on admission, blood lactate > 2.5 mmol/L at 8, 16 and 24 hours after admission, and increase or < 50% decrease in blood lactate within 8 and 16 hours after admission. Blood lactate persistently > 4.4 mmol/L indicated a very poor prognosis. The study concluded that serial blood lactate measurements are useful in predicting survival in dogs with severe and complicated canine babesiosis.
Dissertation (MMed Vet (Med))--University of Pretoria, 2004.
Companion Animal Clinical Studies
unrestricted

Babesiosis and anaplasmosis are part of the complex called Bovine Parasitic Sadness (TPB), a disease that causes high morbidity and mortality in susceptible cattle. It is caused by infection of Babesia bovis and Babesia bigemina protozoa and by the bacterium Anaplasma marginale. The objective of this study was to evaluate oxidative stress parameters in cattle experimentally infected with B. bovis and B. bigemina and the activities of cholinesterase in asymptomatic B. bigemina well as interference of splenectomy in the oxidative balance of cattle infected with A. marginale. For this, three experiments were performed, using 24 young cattle divided into three groups, each consisting of eight animals which: in the first experiment, four control and four infected with attenuated strain of B. bovis, which were observed decrease in erythrocytes count and activities of catalase (CAT) and superoxide dismutase (SOD) in addition to increased levels of thiobarbituric acid reactive substances (TBARS). In Experiment II, we used four cattle control and four infected with attenuated strain of B. bigemina, observed decrease in the activities of acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and CAT, and increased levels of TBARS and SOD in infected cattle. And, in Experiment III, four splenectomized cattle and four intact, both groups infected with A. marginale, with no difference between groups in the hematological and enzymatic profile, only observed drop in hematocrit, red blood cell count and hemoglobin concentration, and increased total leukocyte count due to lymphocytosis in both groups. Positive correlations were found between TBARS versus bacteremia and negative between NPSH versus bacteremia in both groups, but the correlations were higher in splenectomized group. From the results it can be inferred that infection with B. bovis causes oxidative balance, in the same way B. bigemina infection induces an oxidative stress condition and changes the atividase cholinesterase even in asymptomatic animals and bacteremia by A. marginale influences lipid peroxidation in independent splenectomy cattle. This study may suggest that oxidative stress and low-grade inflammation markers can be used as auxiliary tool in the early diagnosis of this disease as well as the basis for studies on the use of antioxidants in the diet of cattle to prevent infection and / or reduce the severity of injuries caused by these parasites.
A Tristeza Parasitária Bovina (TPB) é uma doença que causa alta morbidade e mortalidade em bovinos suscetíveis, causada pela infecção dos protozoários Babesia bovis e Babesia bigemina e pela bactéria Anaplasma marginale. O objetivo deste estudo foi avaliar parâmetros de estresse oxidativo em bovinos experimentalmente infectados com B. bovis e B. bigemina e as atividades das colinesterases na infecção assintomática por B. bigemina assim como a interferência da esplenectomia no equilíbrio oxidativo de bovinos infectados com A. marginale. Para tanto, foram realizados três experimentos, sendo utilizados 24 bovinos jovens divididos em três grupos experimentais, cada um composto por oito animais sendo: no Experimento I, quatro controles e quatro infectados com cepa atenuada de B. bovis, onde foram observados decréscimo na contagem de hemácias e nas atividades das enzimas catalase (CAT) e superóxido dismutase (SOD) concomitantes com aumento nos níveis das substâncias reativas ao ácido tiobarbitúrico (TBARS). No experimento II, foram usados quatro bovinos controles e quatro infectados com cepa atenuada de B. bigemina, sendo observados decréscimo nas atividades das enzimas acetilcolinesterase (AChE), butirilcolinesterase (BChE) e CAT e aumento nos níveis de TBARS e SOD nos bovinos infectados. E, no Experimento III, quatro bovinos esplenectomizados e quatro intactos, ambos os grupos infectados com A. marginale, não havendo diferença entre os grupos no perfil hematológico e enzimático, apenas observada queda no hematócrito, contagem de hemácias e concentração de hemoglobina e aumento na contagem total de leucócitos devido a um aumento na contagem de linfócitos em ambos os grupos. Foram evidenciadas correlações positiva entre TBARS e a bacteremia e negativa entre NPSH e a bacteremia em ambos os grupos, porém as correlações foram maiores no grupo esplenectomizado. A partir dos resultados pode-se inferir que a infecção por B. bovis causa desequilíbrio oxidativo, da mesma forma que a infecção por B. bigemina induz a uma condição de estresse oxidativo e altera a atividade das colinesterases mesmo em animais assintomáticos e que a bacteremia por A. marginale influencia na peroxidação lipídica em bovinos independente da esplenectomia. Com este estudo, pode-se sugerir que marcadores de estresse oxidativo e de inflamação de baixo grau podem ser utilizados como ferramenta no auxílio do diagnóstico precoce desta enfermidade assim como servir de base para estudos referentes ao uso de antioxidantes na alimentação de bovinos para prevenir a infecção e/ou reduzir a gravidade das lesões causadas por estes parasitas.

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior
Babesiosis, anaplasmosis, trypanosomosis and borreliosis are relevant diseases, potentially causing morbidity in cattle, leading to economic losses. The objective of this study was to determine, through indirect enzyme-linked immunosorbent assay (ELISA), the prevalence of Babesia bigemina, Babesia bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia sp. antibodies in cattle to estabilish the risk of outbreaks. Sera samples of 246 female adult cattle from Castanhal and S?o Miguel do Guam?, northeastern Par? counties (areas with recorded copper and cobalt deficiency) were used. Crude antigen was used in the serologic technique to detect antibodies against B. bigemina, B. bovis, T. vivax and Borrelia sp. For the detection of A. marginale antibodies, recombinant major surface 1a protein (MSP1a) was used as antigen for indirect ELISA. The serum samples showed a prevalence of 99.2% (B. bigemina), 98.8% (B. bovis), 68.3% (A. marginale), 93.1% (T. vivax) and 54.9% (Borrelia sp). The municipalities were considered stable enzootic areas for B. bigemina, B. bovis and T. vivax, but unstable ones for A. marginale and Borrelia sp. In spite of the stability determined for some studied areas, babesiosis, anaplasmosis and trypanosomosis cases with severe clinical signs were seen on the studied region, possibly due to the mineral deficiency that decreases the immune response and immunoreativity of the animals.
A babesiose e a anaplasmose juntamente com a tripanossomose e borreliose s?o doen?as de import?ncia por causarem, potencialmente, morbidade no rebanho bovino, causando preju?zos ? economia. O presente trabalho teve como objetivo investigar, atrav?s do teste ELISA indireto, a preval?ncia de anticorpos para Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax e Borrelia burgdorferi em bovinos, avaliando o risco de surtos de doen?a. Foram utilizadas amostras de soro de 246 bovinos f?meas, adultas, provenientes dos munic?pios de Castanhal e S?o Miguel do Guam?, regi?o nordeste do estado do Par? com registros de defici?ncias de Cobre e Cobalto. Foi feita sorologia atrav?s do ensaio imunoenzim?tico ELISA indireto, utilizando-se ant?genos brutos para os agentes: B. bigemina, B. bovis, T. vivax e B. burgdorferi. Em rela??o a A. marginale, foi utilizada a prote?na recombinante MSP1a como ant?geno. A an?lise dos soros revelou a preval?ncia para B. bigemina de 99,2%, B. bovis 98,8 %, A. marginale 68,3%, T. vivax 93,1% e Borrelia sp 54,9%. Os munic?pios estudados foram caracterizados como enzooticamente est?veis para B. bigemina, B. bovis e T. vivax, e enzooticamente inst?veis para A. marginale e Borrelia sp. Apesar da estabilidade prevista para alguns agentes na ?rea do estudo, casos cl?nicos de babesiose, anaplasmose e tripanossomose ocorrem na regi?o, possivelmente devido ? defici?ncia mineral, que leva a diminui??o da resposta imune e da imunorreatividade org?nica dos animais.

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior
This work was carried out at Cavalry School Regiment of Brazilian Army to analyze clinical occurrence of babesiosis caused by Babesia equi. There were been used half-breed horses with average age of 11 years old, apparently healthy. These horses received handling standard of Military Unit. Biochemistry methods were used to determinate plasmatic proteins, bilirubins, alkaline phosphatase and lactate dehydrogenase and these tests were correlated with occurrence of B. equi at optical microscopy, horse s age and handling, and antibodies against B. equi using indirect immunofluorescence reaction. It s can be concluded that positive serology to babesiosis didn t affected significantly horses body score. Horses handling and lesser laboratorial tests favors Babesiosis by B. equi. Prevalence of B. equi detected by indirect Immunofluorescence Reaction was 90,24%, similar to others works conducted at Rio de Janeiro State, and can be considered an endemic disease. Horses were on a stable enzootic situation and so B. equi can be observed at optical microscopy relatively higher on stabled horses, because they were under stressing conditions, always exposed to return to disease. Plasmatic levels of alkaline phosphatase confirm clinically that horses presented osteopathies and occurrence of hyperglobulinemia and direct bilirrubinemia indicated hepatic damage in horses.
Este trabalho foi realizado no Regimento Escola de Cavalaria do Ex?rcito Brasileiro, com o objetivo de avaliar a ocorr?ncia cl?nica de babesiose por Babesia equi. Foram utilizados eq?inos sem ra?a definida, com a m?dia de idade de 11 anos e aparentemente sadios. Os eq?inos receberam o manejo padr?o da Unidade Militar. Foram executadas as t?cnicas bioqu?micas para a dosagem das prote?nas, bilirrubinas, fosfatase alcalina e lactato desidrogenase, com a finalidade de avaliar os valores destes com o esfrega?o sang??neo, o manejo, a idade, a sorologia para a detec??o de anticorpos anti-Babesia equi por imunofluoresc?ncia indireta. As conclus?es deste trabalho foram as seguintes: a sorologia positiva para a babesiose n?o interfere significativamente no estado corporal dos animais; o tipo de manejo e o reduzido acompanhamento laboratorial propiciam a ocorr?ncia da doen?a provocada pela B. equi. A preval?ncia de 90,24% de B.equi pela rea??o de imunofluoresc?ncia indireta, observada nos eq?inos foi semelhante ? relatada em outros trabalhos realizados no Estado do Rio de Janeiro, o que faz-se considerar o plantel estudado como end?mico para esta doen?a. Os animais encontravam-se em situa??o de estabilidade enzo?tica conseq?entemente, a observa??o da B. equi na microscopia ?ptica foi relativamente alta nos eq?inos aquartelados, indicando condi??es de estresse e o risco de reagudizac?o da doen?a. Os n?veis plasm?ticos da fosfatase alcalina indicam que plantel sofre osteopatias desmineralizantes e, a ocorr?ncia de hiperglobulinemia e bilirrubinemia direta indica de dano hep?tico nos eq?inos.

Canine babesiosis is an emerging infectious disease in the United States (US). An epidemic of Babesia gibsoni infections in the US was identified. An association between dog breed and B. gibsoni infections was detected. Babesia gibsoni-infected dogs were more likely to be American pit bull terriers and B. canis vogeli infected dogs were more likely to be greyhounds. An association between a recent dog bite and B. gibsoni infection was detected, implicating direct dog-to-dog transmission as a route of infection in the US. Several genes from canine Babesia spp. were characterized, including 18S ribosomal RNA (rRNA), internal transcribed spacer regions (ITS), cytochrome B (cytB), and rhoptry-associated protein-1 (RAP-1). These genetic data were used to develop a sensitive and specific diagnostic semi-nested polymerase chain reaction (PCR) test for canine babesiosis. Using this assay, a novel large Babesia organism was identified in a blood sample obtained from a clinical patient. Molecular and phylogenetic characterization of this large Babesia spp. determined that it was most closely related to B. bigemina. Lastly, an atovaquone and azithromycin drug combination was shown to be the first treatment to clear canine B. gibsoni infections.

Para determinação da ocorrência de transmissão transplacentária da Theileria equi em neonatos eqüinos foram avaliados 50 potros da raça Puro Sangue Lusitano, machos e fêmeas, bem como suas respectivas mães, logo após o parto. Foram colhidas amostras de sangue total, tanto das mães como dos neonatos, entre as primeiras cinco horas pós parto para pesquisa de Theileria equi e Babesia caballi através da técnica de RT-PCR. Utilizou-se o kit de detecção baseado no fluofóro intercalante de DNA SYBERgreen. Um total de 46% das éguas apresentaram resultado positivo para Theileira equi e 54% se mostraram negativas, enquanto que 66% dos potros apresentaram resultados positivos e 34% negativos, sendo que 73,9% dos potros positivos nasceram de mães também positivas. Já para Babesia caballi, 16% das éguas foram positivas e 84% negativas, assim como 2% dos potros foram positivos e 98% negativos. O teste de RT-PCR é bastante sensível e específico, mas pode resultar em falso negativo, apesar de ser eficaz na detecção da Theileria equi e Babesia caballi nos eqüinos. Estes dados permitem concluir que existe a possibilidade de transmissão transplacentária de Theileria equi.
The occurrence of transplacentary transmission of Theileria equi in horses was determined by evaluating 50 young male and female horses of the breed Lusitano Horses as well as their respective mothers. Colts and fillies were evaluated as soon as they were born. Total blood samples were collected from both mother and offspring within the first five hours right after the parturition to analyse Theileria equi and Babesia caballi through the RT-PCR technique. It was used the kit of detection based on DNA SYBERgreen. This study showed us that 46% of the female horses had positive results for Theileira equi and 54% negative results while 66% of the male horses had positive results and 34% of them, negative ones. Moreover, 73.9% of the positive young horses also had their mothers positive. However, for Babesia caballi 16% of the female horses had positive results and 84% negative ones while 2% of the male horses had positive results and 98%, negative ones. The RT-PCR test is very sensitive and specific but it can occur false-negative results although it is efficient in detecting Theileria equi and Babesia caballi in horses. In conclusion, the data show us that there is a possibility of transplacentary transmission of Theileria equi.

The pathology of canine cerebral babesiosis was examined at the gross, histological and ultrastructural levels. Gross lesions could be categorised as either global or regional. Congestive brain swelling , diffuse cerebral congestion and diffuse cerebral pallor were classified as global lesions. Multifocal haemorrhage and malacia were classified as regional lesions. Oedema was inconsistently present and could be either focal or diffuse. The majority of histological changes were observed in both cerebral babesiosis and control cases. Regional lesions were unique to cerebral babesiosis and had specific histological features. Highly localised endothelial injury was the primary lesion. Early lesions were multifocal and strictly associated with the microvasculature. Intermediate lesions, with perivascular haemorrhage and neutrophil infiltration, were suggestive of reperfusion injury. Advanced lesions were locally extensive and similar in appearance to haemorrhagic infarction. It is likely that the pathogenesis of regional lesions is by a process of microvascular infarction, as venous thrombosis could not be demonstrated. Ultrastructural evidence for adherent contact between erythrocytes and capillary endothelium was demonstrated. Endothelial cell necrosis occurred early in the development of lesions, before neuronal and glial injury. It is postulated that endothelial injury is the primary event in the development of regional lesions and secondary lesions develop as a consequence of microvascular infarction.
Die patologie van die serebrale vorm van bosluiskoors in honde is ondersoek. Die letsels is makroskopies, histologies en elektronmikroskopies beskryf. Letsels kon makroskopies in twee groepe verdeel word: Globale letsels en gelokaliseerde letsels. Kongestiewe brein swelling, diffuse serebrale kongestie en serebrale anemie kom voor as globale letsels in serebrale babesiose. Multifokale bloeding en nekrose kom voor as gelokaliseerde letsels. Edeem was nie konsekwent teenwoordig nie, en was algemeen of verspreid. Die meeste algemene histologiese veranderinge was in beide serebrale en kontrole gevalle teenwoordig. Gelokaliseerde letsels waarin spesifieke hisotpatologiese veranderinge voorgekom het, was kenmerkend van serebrale babesiose. Die primere letsel is hoogs gelokaliseerde beskadiging van endoteelselle. Beskadiging van die kapillere bloedvate ontstaan vroeg in die ontwikkeling van letsels. Verdere ontwikkeling van die letsel word gekenmerk deur peri-vaskulere bloeding en neutrofiel infiltrasie wat aanduidend is van reperfusie beskadiging. Volontwikkelde letsels is plaaslik-ekstensief en het die voorkoms van hemoragiese infarkte Dit is waarskynlik dat mikrovaskulere infarksie 'n rol speel in die patogenese van die letsels, aangesien veneuse trombose nie ontstaan nie. Noue kontak tussen rooibloedselle en kapillere endoteel is elektronmikroskopies bevestig. Endoteelselnekrose ontstaan voordat tekens van beskadiging geidentifiseer kan word in neurone of gliaselle. Dit blyk dat kapillere endoteelselbeskadiging die primere letsel by die ontstaan van gelokaliseerde lese Is is, en dat sekondere lesels ontwikkel as gevolg van mikrovaskulere infarksie.
Dissertation (MSc)--University of Pretoria, 2000.
Paraclinical Sciences
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Electrocardiographic (ECG) changes have never been reported in canine babesiosis. Based on the metabolic, electrolyte, and myocardial alterations described for the disease, such changes are to be expected. The purpose of this study was to describe ECG changes in canine babesiosis, and to correlate those changes to clinical severity, outcome and cardiac histopathological changes. Four groups of dogs with babesiosis were studied: mild to moderate anaemia (n=40), severe anaemia (n=35), concurrent immune-mediated haemolytic anaemia (n=18) and complicated (n=28). Lead II ECG was recorded at admission for 1 minute in all dogs, and repeated after 24 hours in admitted dogs (groups II – IV). Six lead ECG was recorded in 88 dogs. Full necropsy was performed between 30-60 minutes after death on 16 dogs (5 died on arrival, 11 had ECG recording). Gross cardiac pathology was recorded and histopathology of myocardial sections from ventricles, atria, apex and interventricular septa was evaluated, using a scoring system for haemorrhages, necrosis, inflammatory infiltrate and fibrin microthrombi. The following ECG changes were recorded: sinoatrial (7%) and atrioventricular blocks (4%), ventricular premature complexes (7%), low R-amplitude (23%), prominent Q (33%), axis deviations (40%), prolonged QRS (32%), ST depression and coving (28%), large T (42%), and notched R (28%). Differences between groups were minor and inconsistent. Gross pathological changes were pericardial effusion (25%) and subepicardial (56%) and subendocardial haemorrhages (63%). Histological changes were haemorrhages (69%), necrosis (50%), inflammation (63%) and fibrin microthrombi (75%). The only correlation between pathology and ECG was low R-amplitude and pericardial effusion. There was a significantly higher prevalence of sinus bradycardia and irregular sinus rhythm in the non-survivors. Both ECG and pathological changes were non-specific, but there were similarities to the pattern of changes that have been described for myocarditis and myocardial ischaemia. Antiarrhythmic treatment was only required in 1 dog. Thus, the clinical application of the ECG changes found in this study was limited. It was concluded that the heart suffers from the same pathological processes described in other organs in canine babesiosis, namely inflammation and hypoxia. Cardiovascular management, if necessary, should be based on functional monitoring rather than ECG.
Dissertation (MMedVet (Med))--University of Pretoria, 2001.
Companion Animal Clinical Studies
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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, Brasil.
This work describes the dairy farms profile and of its owners, as well as the epidemiological condition of the herd to Babesia bovis, Babesia bigemina and Anaplasma marginale infection in the south mesorregi?o Espirito-santense, with base in a serologic study. Owners or manager of 38 dairy farms were interviewed with a semi-structured questionnaire and the data analyzed by descriptive statistics. In 97% of the dairy farms, the herd was formed by animals with different degrees of blood girolando. Of all producers, 61% were exclusively dairy farms owners and 34% were also coffee producer. The extensive creation system is used in 61% of dairy farms. The dairy farms management, in 89% and 11% of the cases were accomplished by the owners and administrators, respectively. The serologic study was accomplished using indirect ELISA of 756 samples of bovine serum, 380 of dairy cows and 376 of calves in breast feeding phase. The mesorregi?o studied was classified in situation of stability for B. bovis (98,4%), B. bigemina (96,7%) and A. marginale (96,6%) to dairy cows. In relation to the calves was observed prevalence of 75,5% to Babesia bovis, 78,5% to B. bigemina and 69,2% to A. marginale. In general, the results demonstrated that is possible to increase the production and productivity of these dairy farms, but is necessary the owners adapts to the current competitive productive milk system, mainly about the managerial aspect. In the south mesorregi?o Esp?rito-santense the risk of occurrence of outbreaks of babesiosis or anaplasmosis in adult animals is low. However, preventive measured should be adopted for the calves, mainly for those with less to three months of age.
Este trabalho descreve o perfil de propriedades leiteiras e dos produtores, bem como a situa??o epidemiol?gica do rebanho da mesorregi?o sul Esp?rito-santense em rela??o ? Babesia bovis, Babesia bigemina e Anaplasma marginale, com base em um estudo sorol?gico. Propriet?rios ou administradores de 38 estabelecimentos produtores foram entrevistados com uso de um question?rio semi-estruturado e os dados analisados atrav?s de estat?stica descritiva. Em 97% das propriedades visitadas, o rebanho leiteiro era formado por animais com diferentes graus de sangue girolando. Dentre os produtores, 61% se dedicavam exclusivamente a produ??o leiteira e 34% tamb?m se dedicavam ? cafeicultura. O sistema de cria??o extensiva foi utilizado em 61% das propriedades. Foi observado que o gerenciamento, em 89% e 11% dos casos era realizado pelos propriet?rios e administradores, respectivamente. O estudo sorol?gico foi realizado utilizando o m?todo de ELISA indireto a partir de 756 amostras de soro bovino, dos quais 380 de vacas em lacta??o e 376 de bovinos jovens em fase de aleitamento. Para vacas em lacta??o, a mesorregi?o foi classificada em situa??o de estabilidade para B. bovis (98,4%), B. bigemina (96,7%) e A. marginale (96,6%). Em rela??o aos bovinos jovens foi observada preval?ncia de 75,5% para Babesia bovis, 78,5% para B. bigemina e 69,2% para A. marginale. Os resultados demonstraram que, em geral, as propriedades possuem potencial para aumentar sua produ??o e produtividade, mas os produtores precisam se adequar ao competitivo sistema de produ??o de leite, principalmente no aspecto gerencial. Na mesorregi?o sul Esp?rito-santense o risco de ocorr?ncia de surtos das doen?as do Complexo Tristeza Parasit?ria Bovina ? baixo para os animais adultos. Contudo, medidas preventivas devem ser adotadas para os bezerros, principalmente para aqueles com menos de tr?s meses de idade.

Babesiosis, caused by Babesia rossi, is a common cause of morbidity and mortality of dogs in South Africa. Canine babesiosis can be classified either as uncomplicated or complicated based on the degree of anaemia and the severity of the presenting clinical signs.1,2 In uncomplicated babesiosis, the clinical signs are mostly attributable to the degree of the anaemia, whereas in complicated babesiosis the disease process is characterised by additional organ involvement.3,4 One of the most common haematological hallmarks of canine babesiosis, caused by B. rossi, is thrombocytopenia, which is not associated with clinical haemorrhage despite very low platelet counts that would normally cause inability to maintain normal primary haemostatic function.5 The aim of this study was to describe the thromboelastographic findings in uncomplicated canine babesiosis and compare them with those of normal, healthy control dogs. We hypothesised that these dogs would have a normal to hypercoagulable haemostatic capacity, despite the severe thrombocytopenia, and that this could be detected with thromboelastography (TEG), which has previously been shown to correlate well with clinical signs of haemorrhage in dogs.6 This was a prospective, cross sectional, observational study that included 20 client-owned dogs, diagnosed with uncomplicated canine babesiosis at the Onderstepoort Veterinary Academic Hospital (OVAH). Infection with B. rossi was confirmed by polymerase chain reaction (PCR) and reverse line blot (RLB) hybridisation assay. Blood samples were collected at the time of diagnosis. A group of 10 healthy control dogs were included for comparison. Antithrombin activity (AT) was measured using an automated spectrophotometric analyser (Cobas Integra 400, Roche, South Africa). D-dimer was measured using an immunometric flow-through principle (D-dimer Single test, Nycocard Reader II, Medinor A/S). Prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen assays were performed on the ST art® 4 analyser (Diagnostica Stago, Roche, South Africa). TEG analysis was performed using the TEG® 5000 Thromboelastograph® Haemostasis System (Haemoscope, Pro-Gen Diagnostics (Pty) Ltd, South Africa). A complete blood count was performed on the ADVIA 2120 (Siemens, South Africa). The results of the babesiosis and control groups were compared using the Mann-Whitney U test or the Students t-test based on normality. The normality assumption for distribution of the variables in the data was evaluated using the Shapiro-Wilk test. The statistical significance was set at p<0.01. The mean haematocrit (Ht) and median platelet count was significantly lower in the babesiosis group than the controls (0.29 vs. 0.50 L/L; p<0.01 and 22.0 vs. 374.5 x 109/l; p<0.01, respectively). There was no significant difference in any of the TEG parameters between the babesiosis group and the controls. The medians for the various TEG parameters in the babesiosis group versus the controls were; R: 5.5 vs. 4.4 min (p=0.05); K: 2.5 vs. 2.0 min (p=0.08); angle: 58.3 vs. 61.1 degrees (p=0.35); MA: 47.0 vs. 57.0 mm (p=0.02); G: 4.9 vs. 6.7 dyn/cm2 (p=0.02); LY30: 0.00 vs. 0.6% (p=0.20); and LY60: 0.00 vs. 3.0% (p=0.014). The median fibrinogen concentration was significantly higher in the babesiosis group than in the control group; 5.8 g/L (5.0 – 7.0) vs. 2.9 g/L (2.5 – 3.3); (p<0.01). The mean AT activity was significantly lower in the babesiosis group than in the control group; 102.6 mg/dl (89.9 – 112.8) vs. 127.8 mg/dl (110.6 – 134.8); (p<0.01). The median D-dimer concentration was not significantly different in the babesiosis group compared to the control group; 0.3 mg/L (0.1 – 0.4) vs. 0.1 mg/L (0.1 – 0.2); (p=0.016). Median PT was not significantly different in the babesiosis group compared to the control group; 6.5 sec (6.4 – 7.2) vs. 6.8 sec (6.6 – 7.5); (p=0.14). Median aPTT was significantly prolonged in the babesiosis group compared to the control group; 13.6 sec (12.4 – 14.5) vs. 11.5 sec (10.7 – 12.2); (p<0.01). Despite the severe thrombocytopenia, dogs suffering from uncomplicated babesiosis did not have clinical signs of haemorrhage. The thromboelastograms of the babesiosis group were normal to hypercoagulable and thus correlated well with the clinical phenotype. Copyright
Dissertation (MSc)--University of Pretoria, 2011.
Companion Animal Clinical Studies
unrestricted

The genus Babesia comprises intraerythrocytic protozoa that cause babesiosis disease with flu-like symptoms in young immunocompetent individuals that can develop into anaemia, organ failure, neurological complications, and even death in older or immunocompromised patients. Babesia spp. are known to infect both humans and a wide range of wild and domestic animals. More than 100 species of Babesia have been identified but only a few species are known to cause disease in humans. Babesiosis causes huge economic losses in the cattle industry, with 1.2 billion cattle at risk of Babesia infection worldwide. In Australia alone, this disease costs the cattle industry millions of dollars, not just as a result of the mortality and abortion in infected cattle, but also due to the costs of control measures such as acaricidal baths and drug administration. Current control measures for bovine babesiosis include chemotherapeutics, acaricides, and live vaccines, which have inherent limitations and are non-sustainable, while no human vaccine is currently available to protect against human babesiosis. Effective, safe and sustainable vaccines are needed. In this thesis, novel approaches were taken to develop whole Babesia microti (King strain) parasite vaccines. In Chapter 3 the B. microti parasitised red blood cell (pRBCs) dose was characterised in a rodent model using two strains of mice (BALB/c and C56BL/6) and the dose of B. microti pRBCs was optimised for two different inoculation routes. We noted that BALB/c mice suffered from recrudescence, while C56BL/6 cleared parasites and no recrudescent parasitemia peak was detected. The findings in Chapter 3 were used in whole blood-stage parasite vaccine studies in the subsequent chapters. We subsequently examined whether a chemically attenuated B. microti pRBCs vaccine, where the parasite is treated in vitro with the chemical agent, tafuramycin-A, that previously induced protective immunity in pre-clinical vaccine studies of the related malaria parasite, could protect the vaccinee. We showed that mice intravenously administered B. microti pRBCs that had been treated with 2μM tafuramycin-A in vitro, were protected against homologous challenge. We then immunised mice with a threedose regimen of a chemically attenuated whole blood stage B. microti vaccine and showed protection with prolonged immunological memory at 9 months after the last immunisation. Protection was shown to be CD4+ T-cell-dependent. Analysis of the protective efficacy is described in Chapter 4. A live attenuated B. microti vaccine has limitations associated with storage, and delivery considerations. An alternative strategy is a liposome-based antigen delivery platform, whereby liposomes are used to deliver antigens to target cells. In Chapter 5, we produced freshly prepared mannosylated/ unmannosylated liposomes containing parasite extract of B. microti pRBCs using the thin-film hydration method as described in Chapter 2 and examined in a three-dose regimen their protective efficacy against homologous challenge with B. microti pRBCs compared with the chemically attenuated B. microti pRBCs vaccine in a rodent model. We then examined freshly prepared mannosylated liposomes containing parasite extract of B. divergens pRBCs, and studied induced immune responses and protective efficacy against heterologous challenge. Furthermore, we prepared lyophilised mannoyslated liposomes containing B. microti as described in Chapter 2 and examined their protective efficacy and induced immune responses to those freshly prepared containing B. microti against homologous challenge in order to address some of the difficulties associated with the storage and delivery of current whole blood-stage parasite vaccine approaches. We then compared in the protective efficacy and immune responses between lyophilised mannoyslated liposomes containing B. microti or B. divergens pRBCs against homologous or heterologous challenge. Stability of lyophilised vaccine stored at 4°C for a month prior to initiation of the immunisation regimen was evaluated and found to protect against homologous challenge. Lyophilised mannosylated liposomes containing B. microti pRBCs induced immunity with immunological memory at 3 months after the last immunisation. This protection was again determined to be CD4+ T-cell-dependent. Analysis of the protective efficacy is described in Chapter 5. We present a preclinical study employing a novel strategy of live whole organism (chemical attenuated B. microti vaccine) and non-live mannosylated liposomal vaccines to combat babesiosis. The data obtained further our understanding of the immune responses elicited following immunisation with both chemically attenuated B. microti and mannosylated liposome vaccines containing whole parasite antigens. Vaccination with freshly prepared mannosylated liposomes and lyophilised vaccines containing B. divergens antigens induces cross-species protection against B. microti, and as such this strategy could be employed for producing a vaccine from cultured B. divergens parasites. Lyophilised mannosylated liposomes provide a feasible strategy towards practical vaccine storage for use in the field. These data provide strong grounds towards further investigation of an effective babesial vaccine in a clinical setting.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Institute for Glycomics
Griffith Health
Full Text

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FUNDAÇÃO DE AMPARO À PESQUISA E AO DESENVOLVIMENTO CIENTIFICO E TECNOLÓGICO DO MARANHÃO
Canine babesiosis and canine visceral leishmaniasis (CVL) are vector borne diseases, where dogs exert a play as reservoir or source for arthropods responsible by the transmission of these protozoosis. Babesia canis vogeli, is transmited by tick Rhipicephalus sanguineus while Leishmania infantum (sin. Leishmania chagasi) is transmited by sand fly (Lutzomyia longipalpis). The results showed that CVL remains endemic in São Luís Municipality. Despite that the coinfection between Leishmania and Babesia was low considering that both diseases are endemic in this tropical area. Beside that was observed that Yorkshire terrier presented higher predisposition to acquire the infection by B. canis vogeli. A remarkable result was the occurrence of ocular lesions associated to L. infantum infection.
A babesiose canina e a leishmaniose visceral canina são doenças transmitidas por vetores, sendo os cães competentes reservatórios e fonte alimentar dos artrópodes envolvidos. Babesia canis vogeli, é um parasita intraeritrocitário transmitido pelo carrapato Rhipicephalus sanguineus. Leishmania infantum (sin. Leishmania chagasi) transmitidos a mamíferos pela picada dos flebotomíneos (Lutzomyia longipalpis), infectando macrófagos do Sistema Fagocítico Mononuclear do hospedeiro. O presente trabalho é divido em capítulos e os resultados encontrados mostraram que a leishmaniose visceral canina ainda é endêmica no município de São Luís, e que apesar disso a taxa de coinfecção com Babesia foi baixa. Também foi observado que a raça Yorkshire terrier, dentre as raças estudadas, apresentou maior predisposição para contrair a infecção por B. canis vogeli. Outro resultado significativo foi a ocorrência de lesões oculares associadas à infecção por L. infantum.

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq
Babesiosis is a hemolytic tick-borne disease caused by intraerythrocytic protozoal parasites of the genus Babesia. This is a disease of high incidence in the South region of Brazil and responsible for considerable economic losses. Clinical-pathological diagnosis can be made by demonstrating the parasite by examining blood smears or fresh tissues. However, frequently, only formalin-fixed organs are sent to pathology laboratories, which makes it impossible to perform the smears. The main objective of this study was to find alternative histochemical techniques capable to improve the identification of intraerythrocytic B. bovis in histological sections. For this study, tissue samples from 50 bovine necropsy cases were retrospectively analyzed, which had been fixed in formalin and routinely processed for histopathology. The cases were divided into a control group, consisting of 12 cases of cerebral babesiosis with characteristic pink-cherry gray matter (group A), and 38 cases suggestive of cattle tick fever (group B), according to necropsy protocols. Histochemical techniques of Alcian Blue, Methylene Blue, Toluidine Blue, Giemsa, Gram (McCallum-Goodpasture method), Grocott, Periodic Acid of Schiff and Ziehl-Neelsen were tested. Among these, it was observed that the techniques of Methylene Blue and Toluidine Blue allowed to observe morphological and dye characteristics in a clearer way, aiding in the identification of B. bovis. In addition, several clinical-epidemiological and anatomopathological parameters of babesiosis caused by B. bovis were established.
A babesiose é uma doença hemolítica transmitida por carrapatos e causada por protozoários intraeritrocitários do gênero Babesia. Esta é uma doença de incidência elevada na Região Sul do Brasil e responsável por perdas econômicas consideráveis. O diagnóstico clínico-patológico pode ser feito através da demonstração do parasita pelo exame de esfregaços sanguíneos ou de tecidos frescos. Entretanto, com frequência, somente órgãos fixados em formol são remetidos para laboratórios de patologia, o que impossibilita a realização do esfregaço. O principal objetivo deste estudo foi encontrar técnicas histoquímicas alternativas e capazes de aprimorar a evidenciação de B. bovis intraeritrocitária. Para este estudo, foram analisadas, retrospectivamente, amostras de tecidos de 50 casos de necropsias de bovinos, as quais haviam sido fixadas em formol e processadas rotineiramente para histopatologia. Os casos foram divididos em um grupo controle, constituído por 12 casos de babesiose cerebral com substância cinzenta encefálica róseo-cereja característica (grupo A), e 38 casos sugestivos de tristeza parasitária bovina (grupo B), conforme os protocolos de necropsias. Foram testadas as técnicas histoquímicas de Azul Alciano, Azul de Metileno, Azul de Toluidina, Giemsa, Gram (método de McCallum-Goodpasture), Grocott, Ácido Periódico de Schiff e Ziehl-Neelsen. Dentre estas, observou-se que as técnicas de Azul de Metileno e Azul de Toluidina permitiram observar características morfológicas e tintoriais de maneira mais nítida, auxiliando na identificação de B. bovis. Adicionalmente, foram estabelecidos vários parâmetros clínico-epidemiológicos e anatomopatológicos da babesiose por B. bovis.

This thesis showed that dogs with natural infection with B. canis had both renal and cardiac dysfunction, both of which can be classified as complications of babesiosis and would thus necessitate supportive therapy. This thesis demonstrated that RTE celluria, proteinuria, and variable enzymuria and azotaemia occur in dogs with babesiosis. However, these were all minimal changes and all could be consistent with hypoxia, reduced GFR, or reduced cardiac output This thesis showed that dogs with naturally occurring babesiosis had significant urine met-haemoglobin with no evidence of blood met-haemoglobin. The possibility would be that the urinary methaemoglobin was either produced in the kidney or possibly by oxidation of haemoglobin to met-haemoglobin in the bladder. It has been shown experimentally that met-haemoglobin can be toxic. The combination of reduced GFR, anaemic hypoxia, and met-haemoglobin can all act synergistically to cause renal damage. Renal haemodynamics are also much more likely to be abnormal when cardiac dysfunction is present Reduced renal blood flow and glomerular filtration rate are evidence of redistribution of blood flow that commonly occurs in early heart failure. An important finding in this thesis was that dogs with babesiosis had lower serum sodium than control dogs but there was no difference between mild, severe, or complicated cases of babesiosis. In addition, dogs with babesiosis had a lower fractional clearance of sodium than Clinically healthy control dogs, which can be interpreted as sodium retention by the kidneys. This sodium retention would also result in water retention , which will result in an expansion of the plasma volume. In the past heart lesions in canine babesiosis were regarded as rare complications, with the majority of lesions being reported as incidental findings at post-mortem examination of complicated babesiosis cases. This thesis has demonstrated that cardiac lesions to be common in canine babesiosis. This thesis showed that that ECG changes in babesiosis were similar to the pattern described for myocarditis and myocardial ischaemia, and together with the histopathological findings indicated that the heart suffers from the same pathological processes described in other organs in canine babesiosis, namely inflammation and hypoxia. As the clinical application of the ECG changes found in this thesis was limited, cardiovascular assessment should be based on functional monitoring rather than ECG. Using cardiac troponin as a marker of myocardial injury, this thesis showed that myocardial cell injury occurs with canine babesiosis. Cardiac troponins, especially troponin I, are sensitive markers of myocardial injury in canine babesiosis, and the magnitude of elevation of plasma troponin I concentrations appears to be proportional to the severity of the disease. ECG changes and serum cardiac troponin were correlated with histopathology. On cardiac histopathology from dogs that succumbed to babesiosis, haemorrhage, necrosis, inflammation and fibrin microthrombi in the myocardium were documented, all of which would have resulted in ECG changes and elevations in cardiac troponin. Myocardial infarction causes left ventricular failure, which will result in hypotension and an expansion of the plasma volume due to homeostatic mechanisms. This thesis showed that dogs with babesiosis had hypoalbuminaemia, which may be because of intravascular volume dilution due to fluid retention. In disease hypoalbuminaemia can occur as a negative acute-phase protein. In the light of the cardiac changes, hyponatraemia, and hypotension, a probable cause would be fluid retention due to myocardial disease. This thesis showed that dogs with babesiosis had left ventricular lesions, which can result in systolic heart failure.
Thesis (PhD)--University of Pretoria, 2005.
Veterinary Tropical Diseases
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In the genera Babesia, Ehrlichia and are positioned Hepatozoon species of parasites responsible for babesiosis, ehrlichiosis and hepatozoonosis respectively. These organisms have in common the fact that they are transmitted by tick vectors in dogs infected and cause general symptoms such as fever, anemia and jaundice accompanied by intracellular infections. In the state of Goias had not work with phylogenetic approach for these blood parasites. The overall objective of this study was molecular analysis of isolates of Babesia, Hepatozoon and Ehrlichia obtained from symptomatic dogs in Goiânia, Goiás State, thus confirming that the species and subspecies involved in infections. For this, DNA extraction was carried out of samples, followed by carrying out PCR with generic primers. After the PCR was obtained fragments of the 18S rRNA region for samples of Babesia and Hepatozoon and 16S rRNA for the detection of species of the genus Ehrlichia. PCR products obtained were purified and used for sequencing. We sequenced 35 samples of Babesia spp. and of these 17 were used for phylogenetic studies. Two samples of Hepatozoon spp. 17 samples were sequenced and Ehrlichia spp which only five were used for analysis. Analyses performed with the identity of sequenced samples allowed the identification of B. vogeli canis, Hepatozoon canis and Ehrlichia canis. When confronted with the species and subspecies of reference from other regions of Brazil and the world showed close molecular similarity, which demonstrates the low variability between different samples from different geographic regions. For the analysis using the program MEGA4 and subsequent construction of a phylogenetic tree samples in this study formed their own groups always association with the reference samples for the respective species. Thus it was concluded that samples of B. c. vogeli H. canis and E. canis from dogs in the city of Goiania GO show close similarity with isolates from other regions of the world
Nos gêneros Babesia, Ehrlichia e Hepatozoon são posicionadas as espécies de parasitos responsáveis pela babesiose, ehrlichiose e hepatozoonose respectivamente. Estes microrganismos têm em comum o fato de serem transmitidos por carrapatos vetores e causarem nos cães infectados sinais gerais como febre, anemia e icterícia acompanhados de infecções intracelulares. No estado de Goiás não haviam trabalhos com abordagem filogenética relativos a estes hemoparasitos. O objetivo geral do presente trabalho foi realizar análise molecular de isolados de Babesia, Ehrlichia e Hepatozoon obtidos de cães sintomáticos na cidade de Goiânia, Estado de Goiás, verificando assim qual a espécie e subespécie envolvida nas infecções. Para isto, foi realizado extração de DNA das amostras, seguido da realização de PCR com oligonucleotídeos genéricos. Após a PCR obteve-se fragmentos da região do 18S rRNA para as amostras de Babesia e Hepatozoon e do 16S rRNA para a pesquisa de espécies do gênero Ehrlichia. Os produtos de PCR obtidos foram purificados e utilizados para o sequenciamento. Foram seqüenciadas 35 amostras de Babesia spp. e destas 17 foram utilizadas para os estudos filogenéticos. Duas amostras de Hepatozoon spp. foram seqüenciadas e 17 amostras de Ehrlichia spp sendo que apenas cinco foram utilizadas para as análises. As análises de identidade realizadas com as amostras seqüenciadas permitiram a identificação de B. canis vogeli, Hepatozoon canis e Ehrlichia canis. Quando confrontadas com as espécies e subespécies de referência de outras regiões do Brasil e do mundo apresentaram estreita similaridade molecular, o que demonstra a baixa variabilidade entre as diferentes amostras de diferentes regiões geográficas. Pela análise feita utilizando-se o programa MEGA4 e subsequente construção de árvores filogenéticas as amostras deste estudo formaram grupos próprios sempre em associação com as amostras de referência para as respectivas espécies. Dessa forma foi possível concluir que as amostras de B. c. vogeli, H. canis e E. canis provenientes de cães da cidade de Goiânia-GO apresentam estreita similaridade com isolados de outras regiões do mundo

A babesiose bovina é uma doença transmitida por carraças, que causa elevada morbidade e mortalidade, e provoca consideráveis perdas económicas devido aos esforços para controlar esta doença. As medidas de controlo da babesiose bovina incluem a erradicação ou redução de carraças, correcto diagnóstico, assim como tratamento e vacinação apropriados. Este trabalho tem como objectivo contribuir para um melhor diagnóstico da infecção e a consequente melhoria de algumas das medidas de controlo, bem como identificar e caracterizar genes de proteases utilizados para o desenvolvimento de um método de diagnóstico e estudados como potenciais alvos para fármacos. Para este estudo, foi realizado um trabalho de colaboração em Moçambique, onde foram colhidas amostras de sangue de bovinos naturalmente infectados, em cinco explorações situadas na província de Maputo, no sul do país. Um novo método de detecção molecular por PCR foi desenvolvido e testado utilizando DNA genómico e amostras de campo aleatórias colhidas numa das explorações. Os iniciadores de PCR foram desenhados com base no gene putativo da protease aspártica babesipsina-1 identificado nos genomas de Babesia bigemina e de B. bovis. O novo seminested hot-start PCR foi desenvolvido utilizando a combinação de iniciadores longos de 30 pb de comprimento e uma hot-start polimerase, que permitem teoricamente a utilização de temperaturas de emparelhamento acima da temperatura de melting, impedindo assim a formação de amplificações não específicas, o que aumenta a especificidade do método. O novo seminested hot-start PCR foi avaliado utilizando 117 amostras de campo, e em paralelo com um método amplamente utilizado, o nested PCR. O seminested hot-start PCR neste estudo foi mais sensível que o nested PCR. Com o seminested hot-start PCR, 90% das amostras foram positivas para B. bigemina, e 82% foram positivas para B. bovis. Os resultados sugeriram que a babesiose bovina é comum e endémica em Moçambique, e que a doença se encontra numa situação de estabilidade endémica. O estudo do estado da babesiose bovina em Moçambique, foi então aprofundado, através da análise de amostras de campo aleatórias de mais quatro explorações utilizando o seminested hot-start PCR. Todas as amostras das cinco explorações foram também analisadas utilizando o RLB, e os resultados deste método foram comparados com os dados obtidos pelo seminested hot-start PCR. A detecção de Babesia spp. diferiu significativamente entre os métodos utilizados e os locais de recolha. Com o seminested hot-start PCR, a detecção de B. bigemina nas várias explorações, variou entre 30% e 89%, com uma detecção total de 61%, e a detecção de B. bovis variou entre 27% e 83% com uma frequência global de 53%. Utilizando o RLB, não foi detectado B. bigemina e a detecção de B. bovis variou entre 0% e 17% com uma frequência total de 5,1%. A análise de novas sequências do gene 18S rRNA, revelou que a actual sonda do RLB para B. bigemina não é adequada para a detecção de todos os isolados desta espécie identificados em Moçambique. O seminested hot-start PCR foi portanto mais sensível que o RLB. No entanto, dez espécies diferentes dos quatro Géneros Anaplasma, Babesia, Ehrlichia e Theileria foram detectadas pelo ensaio RLB, e isso demonstra que as infecções múltiplas são comuns em Moçambique. Os resultados deste estudo mostram que a babesiose bovina é comum na província de Maputo, e também que existem alguns locais com baixa prevalência de infecções, e portanto, os resultados sugerem que esta doença não está numa situação de estabilidade endémica na província de Maputo. São agora necessários novos estudos epidemiológicos para confirmar estes resultados. Tem sido demonstrado que as proteases têm papéis essenciais em parasitas protozoários e estão sob estudo como promissores alvos de fármacos. Algumas proteases cisteínicas de parasitas protozoários, são já reconhecidos alvos de fármacos, e encontram-se em validação inibidores específicos para a quimioterapia da leishmaniose, da malária e da tripanossomíase. Neste estudo, o nosso principal interesse na identificação e caracterização de proteases como alvos de fármacos foi portanto nesta classe de proteases. Foram identificados no banco de dados do projecto em curso de sequenciação do genoma de B. bigemina, os genes putativos de proteases cisteínicas em pesquisas por similaridade de sequência, que posteriormente foram comparados com os genes anotados nos genomas completos das espécies de piroplasmas bovinos B. bovis, Theileria annulata e T. parva. Para avaliar os eventos da evolução molecular que ocorreram na família C1 de proteases cisteínicas, foram feitos alinhamentos múltiplos entre os genomas e análises das sequências obtidas destes piroplasmas de importância veterinária. Existem assim, cinco grupos distintos de genes de proteases cisteínicas da família C1 em B. bigemina (5 genes), quatro grupos em B. bovis (4 genes) e seis grupos em Theileria spp. (13 genes). No Género Theileria a evolução molecular ocorreu através da duplicação de genes e da diversificação da sequência das proteínas codificadas por estes genes. Estas importantes diferenças observadas entre os Géneros Babesia e Theileria na família das proteases cisteínicas, podem parcialmente explicar os diferentes mecanismos de infecção destas espécies, em que parasitas Babesia não invadem linfócitos e parasitas Theileria invadem primeiro os linfócitos no hospedeiro vertebrado. A babesipaína-1, uma das proteases cisteínicas identificadas no genoma de B. bigemina, foi expressa como uma proteína de fusão com a glutationa S-transferase (GST) e a respectiva fracção solúvel foi purificada por cromatografia de afinidade. A babesipaína-1 recombinante apresentou actividade contra certos péptidos que são substratos típicos de proteases cisteínicas, e foi inibida por um inibidor geral da sua classe, mas o baixo rendimento da purificação da fracção solúvel impediu a sua caracterização adicional. A babesipaína-1 foi então purificada a partir da fracção insolúvel, e a proteína desnaturada foi re-enrolada e activada para produzir uma enzima activa. A análise da actividade da babesipaína-1 revelou propriedades típicas de uma protease cisteínica da família da papaína, incluindo a hidrólise de alguns péptidos que são substratos típicos da família da papaína, um pH ácido óptimo (5.5-6.0), o requisito de um ambiente redutor para ter actividade máxima, e a inibição por inibidores de proteases cisteínicas como o E-64, a leupeptina, o ALLN e a cistatina. Os resultados sugerem que a babesipaína-1 tem um papel no citosol, já que a babesipaína-1 manteve elevada actividade contra substratos a pH 7,5 (83% do máximo), uma característica incomum das proteases cisteínicas de parasitas protozoários. Assim, os resultados deste estudo demonstram que a babesiose bovina é uma infecção comum na província de Maputo em Moçambique, embora a doença não esteja numa situação de estabilidade endémica. Os resultados também sugerem que as proteases cisteínicas de Babesia spp. são alvos promissores para fármacos e consequentemente para o desenvolvimento de um tratamento eficaz para a babesiose bovina. Na sequência destes resultados foi associado um plano de trabalho futuro. Alguns pormenores e resultados deste trabalho podem ser transferidos para outros países, inclusivamente Portugal.
Bovine babesiosis is a tick–borne disease with significant morbidity and mortality, and the economic losses associated to this disease can be considerable. Control measures of bovine babesiosis include the eradication or reduction of ticks, good diagnosis, use of vaccination and correct treatment. The aim of this work was to contribute to a better diagnosis of the infection aiming at the improvement of some of the control measures, as well as to identify and characterize protease genes for the development of a diagnostic method and studied as potential drug targets. For this study, collaborative work was carried out in Mozambique, from where blood samples from naturally infected cattle were collected in five farms located in the Maputo province, south of the country. A new molecular detection PCR method was then developed and tested using genomic DNA and random field samples collected from one farm. Primers were designed based in the babesial aspartic protease putative babesipsin gene identified in the genomes of Babesia bigemina and B. bovis. The new seminested hot-start PCR method was developed using the combination of 30 bp long primers and a hot start polymerase that theoretically allows the use of annealing temperatures above the melting temperatures of the primers and prevents the formation of unspecific amplifications and therefore increases the specificity. The new seminested hot-start PCR method was assayed using 117 field samples in parallel with the widely used nested PCR method. The babesipsin seminested hot-start PCR was in this study more sensitive than the nested PCR. With the seminested hot-start PCR, 90% of the samples were positive for B. bigemina and 82% were positive for B. bovis. The results suggested that bovine babesiosis is common and endemic in Mozambique and that the disease was in an endemically stable situation. The status of bovine babesiosis in Mozambique was then further studied, by testing random field samples from four more farms using the seminested hot-start PCR. All the samples from the five farms were also analysed using the reverse line blot (RLB) assay, and the results were compared with the data obtained by the seminested hot-start PCR. The detection of Babesia spp. differed considerably between methods and locations. Using the seminested hot-start PCR, detection of B. bigemina between farms varied from 30% to 89% with an overall detection of 61%, and detection of B. bovis ranged between 27% and 83% with an overall frequency of 53%. Using the RLB assay B. bigemina was not detected and detection of B. bovis ranged between 0% and 17% with a total frequency of 5.1%. Analysis of new sequences of the 18S rRNA gene revealed that the current B. bigemina RLB probe is unspecific for the detection of all the identified isolates from Mozambique. The seminested hot-start PCR was therefore more sensitive than the RLB assay. Nevertheless, ten different species of the four genera Anaplasma, Babesia, Ehrlichia and Theileria were detected by the RLB assay, and this illustrates that multiple infections are widespread in Mozambique. The results of this study show that bovine babesiosis is common in Maputo province, but there are some locations with low prevalence of infections and therefore the results suggest that this disease is not in an endemically stable situation in Maputo province. Further epidemiological studies are now needed to corroborate these findings. Proteases have been shown to have essential roles in parasitic protozoa and are under study as promising drug targets. Some cysteine proteases of protozoan parasites are now recognized drug targets and specific inhibitors are in validation for chemotherapy of leishmaniasis, malaria, and trypanosomiasis. In this study our focus on the identification and characterization of proteases as drug targets was therefore in this class of proteases. Cysteine protease putative genes were identified by sequence similarity search in the ongoing B. bigemina genome sequencing project database and were compared with the annotated genes from the complete bovine piroplasms genomes of B. bovis, Theileria annulata, and T. parva. Multiple genome alignments and sequence analysis were used to evaluate the molecular evolution events that occurred in the C1 family of cysteine proteases in these piroplasms of veterinary importance. There are five distinct groups of cysteine protease genes of C1 family in B. bigemina (5 genes), four groups in B. bovis (4 genes) and six groups in Theileria spp. (13 genes). Molecular evolution in Theileria occurred through the duplication of genes and sequence diversity. These considerable differences observed in the cysteine protease family between Babesia and Theileria genera, may partially explain why Babesia cannot infect lymphocytes and Theileria infect first lymphocytes in the vertebrate host. One of the identified cysteine proteases in the B. bigemina genome, babesipain-1, was expressed as a fusion protein with gluthatione S-transferase (GST) and the soluble protein was purified by affinity chromatography. The recombinant babesipain-1 showed activity against typical peptide substrates of cysteine proteases, and was inhibited by a general inhibitor of its class, but the low yield of the soluble purification prevented additional characterization. Babesipain-1 was then purified from the insoluble fraction, and the denatured protein was refolded and activated to produce an active mature enzyme. Analysis of the activity of babesipain-1 revealed typical properties of a papain-family cysteine protease, including hydrolysis of typical papain-family peptide substrate, an acidic pH optimum (5.5-6.0), requirement for a reducing environment for maximum activity, and inhibition by standard cysteine protease inhibitors as E-64, leupeptin, ALLN and cystatin. The results suggest that babesipain-1 has a role in cytosol environment, since babesipain-1 retained high activity against peptide substrates at pH 7.5 (83% of maximum), an uncommon feature of cysteine proteases of parasitic protozoa. Thus the results of this study demonstrate that bovine babesiosis is widespread in Maputo province in Mozambique, although the disease is not in an endemically stable situation. The results also suggest that cysteine proteases of Babesia spp. are promising drug targets for the development of an effective treatment of bovine babesiosis. In face of these results a plan for future work is associated. Some aspects and results from this work can be adjusted to other countries, including Portugal.

Abnormal carbohydrate metabolism is a commonly encountered feature of malaria in people, and similar derangements have been detected in veterinary patients with canine babesiosis. Glucose, the major metabolic fuel source, is a key resource in critically ill patients as they mount an immunological response to infection and inflammation. The ability of the individual to effectively mobilise, distribute and utilise glucose is a major determinant of morbidity and mortality. Hypoglycaemia has been identified as a life threatening metabolic complication in almost 20% of severely ill dogs suffering from babesiosis due to Babesia rossi infection. Insulin and glucagon are the primary hormones involved in glucose homeostasis. Insulin lowers blood glucose concentration by facilitating cellular uptake and utilisation of glucose. Hyperinsulinaemia as a result of inappropriate insulin secretion may precipitate hypoglycaemia, and has been identif ed as a cause of hypoglycaemia in human and murine malaria. A similar phenomenon may exist in canine babesiosis. This prospective, cross-sectional, observational study, including 94 dogs with naturally acquired virulent babesiosis, sought to investigate and characterise the relationship between blood glucose concentrations and insulin concentrations in cases of canine babesiosis. Pre-treatment jugular blood samples were collected for simultaneous determination of plasma glucose and insulin concentrations. Animals were retrospectively divided into three groups: hypoglycaemic (plasma glucose concentration < 3.3 mmol/L; n=16), normoglycaemic (3.3-5.5 mmol/L; n=62), and hyperglycaemic (> 5.5 mmol/L; n=16). The median plasma insulin concentrations (IQR in parentheses) for the hypoglycaemic, normoglycaemic and hyperglycaemic groups were 10.7 pmol/L (10.7-18.8 pmol/L), 10.7 pmol/L (10.7-29.53 pmol/L; i.e below the detection limit of the assay), and 21.7 pmol/L (10.7-45.74 pmol/L), respectively. Statistical analysis revealed no significant difference in insulin concentration between the three groups. These results suggest that insulin secretion was appropriately suppressed in these dogs. Only two dogs had elevated insulin concentrations, one of which was hypoglycaemic. The median time since last meal (available for 87 dogs) was 24 hours (IQR 2-4 days), constituting a significant period of illness-induced starvation. We conclude that hyperinsulinaemia is not a cause of hypoglycaemia in virulent canine babesiosis. It is speculated that prolonged fasting due to disease-induced anorexia, in addition to increased glucose consumption, depletion of hepatic glycogen stores, and hepatic dysfunction with impaired gluconeogenesis, may play important roles in the pathophysiology of hypoglycaemia in canine babesiosis.
Dissertation (MMedVet)--University of Pretoria, 2010.
Companion Animal Clinical Studies
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Resumen: En esta tesis doctoral se presentan tres modelos matemáticos que describen el comportamiento de dos enfermedades parasitarias con vector de transmisión; de los cuales dos modelos están dedicados a la Toxoplasmosis donde se explora la dinámica de la enfermedad a nivel de la población humana y de gatos domésticos. Los gatos juegan un papel de agentes infecciosos del Toxoplasma gondii. La dinámica cualitativa del modelo es determinada por el umbral básico de reproducción, R0. Si el parámetro R0 < 1, entonces la solución converge al punto de equilibrio libre de la enfermedad. Por otro lado, si R0 > 1, la convergencia es al punto de equilibrio endémico. Las simulaciones numéricas ilustran diferentes dinámicas en función del parámetro umbral R0 y muestra la importancia de este parámetro en el sector salud. Finalmente la Babesiosis bovina se modela a partir de cinco ecuaciones diferenciales ordinarias, que permiten explicar la influencia de los parámetros epidemiológicos en la evolución de la enfermedad. Los estados estacionarios del sistema y el número básico de reproducción R0 son determinados. La existencia del punto endémico y libre de enfermedad se expone, puntos que dependen del R0, ratificando la importancia del parámetro umbral en la salud publica. Objetivo: Construir modelos matemáticos epidemiológicos aplicados a enfermedades parasitarias (Toxoplasmosis y Babesiosis) con vector de transmisión. Metodología: Para la construcción de los modelos matemáticos epidemiológicos es necesario representar la enfermedad a partir de modelos de flujo, permitiendo ver la dinámica de la población entre los diferentes estadíos de la enfermedad, dichos movimientos son analizados a partir de sistemas dinámicos, análisis matemático y métodos numéricos; con estas herramientas es posible hacer un estudio detallado del modelo, permitiendo calcular parámetros umbrales que dominan la dinámica de la enfermedad y a su vez simular escenarios reales e hipotéticos.
Aranda Lozano, DF. (2011). Modeling of parasitic diseases with vector of transmission: toxoplasmosis and babesiosis bovine [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/11539
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Universidade Estadual Paulista (UNESP)
Com o objetivo avaliar a epidemiologia e a característica molecular da infecção de cães de áreas rurais por Babesia spp., amostras de sangue de 150 animais foram coletadas e analisadas por esfregaço sangüíneo e por PCR. Além disso, os cães foram examinados para investigar possível infestação por carrapatos. Pelo esfregaço sangüíneo de sangue capilar (ponta de orelha) foram detectados três (2%) cães positivos e pela PCR para Babesia spp. 12 (8%) deles foram positivos, com bandas visualizadas em 450 pb. Foram encontrados R. sanguineus e Amblyomma spp. em 36 (24%) dos 150 cães. As espécies de Amblyomma detectadas foram A. cajennense (9/36 - 25%), A. ovale (8/36 - 22,2%) e A. aureolatum (1/36 - 2,7%). Um espécime de Amblyomma (1/36 - 2,7%) não pôde ser identificado especificamente pois se encontrava no estágio de ninfa. Não houve uma associação positiva entre o encontro de R. sanguineus e infecção por B. canis. O seqüenciamento de uma amostra positiva demonstrou identidade com B. canis vogeli já caracterizada no Brasil.
In order to study the parasitologic and morphological characteristics of Babesia spp. infection in dogs from rural areas, 150 animals were examined by blood smears and by PCR. Besides, the dogs were examined to investigate ticks infestation. By the blood smear of capillary blood (ear tip) examination, three (2%) were detected positive and by PCR for Babesia spp.12 (8%) were positive, with bands visualized in 450 bp. Rhipicephalus sanguineus and Amblyomma spp. were found in 36 (24%) of the 150 dogs. The found species of Amblyomma were A. cajennense (9/36 - 25%), A. ovale (8/36 - 22.2%) and A. aureolatum (1/36 - 2.7%). A specimen of Amblyomma (1/36 - 2.7%) was found in the nymph stage and could not be identified. There was not a positive association between the encounter of R. sanguineus and the infection for B. canis. The sequencing of two positive samples demonstrated close identity with B. canis vogeli already characterized in Brazil.

Orientador: Lucia Helena O'Dwyer de Oliveira
Banca: Teresa Cristina Goulart de Oliveira-Sequeira
Banca: Vamilton Álvares Santarém
Resumo: Com o objetivo avaliar a epidemiologia e a característica molecular da infecção de cães de áreas rurais por Babesia spp., amostras de sangue de 150 animais foram coletadas e analisadas por esfregaço sangüíneo e por PCR. Além disso, os cães foram examinados para investigar possível infestação por carrapatos. Pelo esfregaço sangüíneo de sangue capilar (ponta de orelha) foram detectados três (2%) cães positivos e pela PCR para Babesia spp. 12 (8%) deles foram positivos, com bandas visualizadas em 450 pb. Foram encontrados R. sanguineus e Amblyomma spp. em 36 (24%) dos 150 cães. As espécies de Amblyomma detectadas foram A. cajennense (9/36 - 25%), A. ovale (8/36 - 22,2%) e A. aureolatum (1/36 - 2,7%). Um espécime de Amblyomma (1/36 - 2,7%) não pôde ser identificado especificamente pois se encontrava no estágio de ninfa. Não houve uma associação positiva entre o encontro de R. sanguineus e infecção por B. canis. O seqüenciamento de uma amostra positiva demonstrou identidade com B. canis vogeli já caracterizada no Brasil.
Abstract: In order to study the parasitologic and morphological characteristics of Babesia spp. infection in dogs from rural areas, 150 animals were examined by blood smears and by PCR. Besides, the dogs were examined to investigate ticks infestation. By the blood smear of capillary blood (ear tip) examination, three (2%) were detected positive and by PCR for Babesia spp.12 (8%) were positive, with bands visualized in 450 bp. Rhipicephalus sanguineus and Amblyomma spp. were found in 36 (24%) of the 150 dogs. The found species of Amblyomma were A. cajennense (9/36 - 25%), A. ovale (8/36 - 22.2%) and A. aureolatum (1/36 - 2.7%). A specimen of Amblyomma (1/36 - 2.7%) was found in the nymph stage and could not be identified. There was not a positive association between the encounter of R. sanguineus and the infection for B. canis. The sequencing of two positive samples demonstrated close identity with B. canis vogeli already characterized in Brazil.
Mestre