Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-25Bitstream added on 2014-06-13T18:56:53Z : No. of bitstreams: 1 andre_mr_me_jabo.pdf: 427316 bytes, checksum: 366b68e3f9cae8ab0938b9ab1032ffb9 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Poucos relatos têm sido feitos sobre o diagnóstico da erliquiose e babesiose em felinos domésticos e selvagens brasileiros, os quais são baseados diretamente pela presença de mórulas em leucócitos e piroplasmas em eritrócitos, e indiretamente pela detecção de anticorpos anti-Ehrlichia canis. O presente estudo teve como objetivo realizar a detecção molecular de E. canis e B. canis e a presença de anticorpos da classe IgG contra esses hemoparasitas em amostras de sangue e soro, respectivamente. Neste utilizamos 72 felídeos selvagens brasileiros mantidos em cativeiro em algumas instituições e zoológicos. Pela Reação de Imunofluorescência Indireta (RIFI), dezoito (25%) e cinqüenta e três (73,6%) dos 72 animais amostrados foram sororeagentes frente aos antígenos de E. canis e B. canis, respectivamente. Na PCR para E. canis, onze (15,3%) dos 72 animais amostrados foram positivos. Os amplicons foram confirmados por seqüenciamento e o DNA de E. canis encontrado mostrou grande similaridade genética com amostras de E. canis isoladas no Brasil, México, Portugal, Grécia e Taiwan, com 98% de similaridade. Nenhuma das amostras foi positiva na PCR para B. canis. Destaca-se a importância e a primeira detecção molecular de E. canis e presença de anticorpos anti-E. canis e anti-B. canis em felídeos selvagens brasileiros mantidos em cativeiro. / Few are the reports that have been carried out on ehrlichiosis and babesiosis diagnostic in Brazilian domestic and wild felids, which are based directly on the presence of morulae in leucocytes and piroplasms in erythrocytes, and indirectly by detection of antibodies against E. canis. The aim of this study was to detect molecularly E. canis and B. canis and the presence of anti-E. canis and B. canis IgG antibodies in the blood and sera samples, respectively, from 72 Brazilian wild captive felids maintained in some instituitions and zoos. Eighteen (25.0%) and fifty-three (73.6%) out of 72 animals were seroreagent for E. canis and B. canis antigen, respectively, by IFA (Indirect Immunofluorescent Assay). Eleven (15.3%) of the 72 samples were positive for nPCR E. canis. The amplicons were confirmed by sequencing and the E. canis DNA found appeared to be closely related to E. canis samples from Brazil, Mexico, Portugal, Greece and Taiwan with 98% percent identity. None of the 72 samples were positive for B. canis by PCR. This is the first molecular detection of E. canis and presence of seroreactivity for both B. canis and E. canis in Brazilian wild captive felids.

Submitted by Simone Souza (simonecgsouza@hotmail.com) on 2018-05-08T14:13:02Z No. of bitstreams: 1 DISS_2015_Rafael Willian Wolf.pdf: 6351388 bytes, checksum: 2670bd6b5374112e3153c188013fae90 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2018-05-23T12:53:29Z (GMT) No. of bitstreams: 1 DISS_2015_Rafael Willian Wolf.pdf: 6351388 bytes, checksum: 2670bd6b5374112e3153c188013fae90 (MD5) / Made available in DSpace on 2018-05-23T12:53:29Z (GMT). No. of bitstreams: 1 DISS_2015_Rafael Willian Wolf.pdf: 6351388 bytes, checksum: 2670bd6b5374112e3153c188013fae90 (MD5) Previous issue date: 2015-03-13 / Os carrapatos são parasitas que possuem uma grande diversidade de hospedeiros e podem transmitir diversos agentes patogênicos. Esses parasitas são responsáveis pela transmissão de agentes infecciosos como as bactérias dos gêneros Rickettsia e Ehrlichia e também protozoários dos gêneros Hepatozoon e Babesia. Relatos mostram que carrapatos do gênero Amblyomma, Ixodes e Rhipicephalus podem atuar como vetores na transmissão destes agentes. Esses carrapatos, nos estágios imaturos geralmente parasitam pequenos mamíferos terrestres enquanto os adultos parasitam mamíferos de médio e grande porte. Levando em conta a diversidade de pequenos mamíferos terrestres do Pantanal e sendo eles hospedeiros para os carrapatos vetores desses agentes infecciosos, o presente estudo teve como objetivo verificar a ocorrência de infecção por Rickettsia spp., Ehrlichia spp., Anaplasma spp., Hepatozoon spp. e Babesia spp., bem como o parasitismo por carrapatos em pequenos mamíferos terrestres da subregião do Pantanal de Poconé. As coletas foram realizadas em oito habitats distintos onde foram instaladas grades amostrais contendo 100 armadilhas cada. Dos animais coletados foram retiradas amostras de sangue, fígado e baço. A detecção dos agentes foi realizada por meio de Reação em Cadeia pela Polimerase (PCR). Foram coletados 64 indivíduos entre marsupiais e roedores, dos quais dez apresentavam parasitismo por carrapatos, sendo um animal parasitado por Amblyomma cajennense, um por Amblyomma triste e oito por Argasideos. Nas PCRs, um roedor (1,6%) foi positivo para o gênero Anaplasma; três (4,7%) foram positivos para Hepatozoon spp., sendo todos eles roedores e quatro (6,2%) foram positivos para Babesia spp., dos quais três (75%) eram roedores e um (25%) era marsupial. Nenhum animal apresentou resultado positivo para Rickettsia spp. ou Ehrlichia spp. Após o sequenciamento das amostras positivas, observou-se para Hepatozoon um fragmento de 560 pb do gene 18S rRNA, que apresentou 99% de similaridade com Hepatozoon sp. (FJ719819) detectado em marsupial do Chile. Para Babesia observou-se à amplificação de dois fragmentos distintos do gene 18S rRNA, um de 532 pb detectado em roedor e outro de 556 pb detectado em marsupial. Ambos apresentaram similaridade de 95% entre si e foram 97% similares com B. bicornis (AF419313). Os resultados sugerem que novas espécies ou genótipos ainda não descritos dos protozoários dos gêneros Hepatozoon e Babesia estão presentes na área estuda e que os pequenos mamíferos podem estar atuando como hospedeiros para esses agentes na região do Pantanal mato-grossense. / Ticks are parasites of a wide range of hosts that can transmit several pathogens. These parasites are responsible for the transmission of infectious agents such as bacteria of Rickettsia and Ehrlichia genus and also protozoa of Babesia and Hepatozoon genus. Previous reports show that Amblyomma, Ixodes and Rhipicephalus ticks genus can act as vectors in the transmission of these agents. Larvae and nymphs stages of the related ticks usually parasitize terrestrial small mammals while adults prey on large mammals. Taking into account the diversity of small terrestrial mammals of the Pantanal, the present study aimed to verify the occurrence of infection by Ehrlichia spp., Anaplasma spp., Rickettsia spp., Hepatozoon spp., Babesia spp. and parasitism by ticks in terrestrial small mammals of the Pantanal of Poconé region. Samples were collected in eight different habitats where sampling grids were installed containing 100 traps each. Samples of blood, liver and spleen were collected from animals captured. The detection of agents was performed by the Polymerase Chain Reaction (PCR). Sixty-four animals, marsupials and rodents were collected, and ten presented parasitism by ticks, one parasitized by Amblyomma cajennense, one by Amblyomma triste and eight by Argasidae. In the PCR analysis, one rodent (1.6%) was positive for the genus Anaplasma; three rodents (4,7%) were positive for Hepatozoon spp. and four (6,2%) were positive for Babesia spp., three of them (75%) were rodents and one (25%) was a marsupial. No animal presented positive result for Rickettsia spp. or Ehrlichia spp. After the sequencing analysis of positive samples, there was observed a fragment of 560 pb of 18S rRNA of Hepatozoon showed 99% of similarity with Hepatozoon sp. (FJ719819) detected in a marsupial species from Chile. For Babesia two distinct fragments of the 18S rRNA gene were amplified, one of 532 pb detected in a rodent and one of 556 pb detected in a marsupial. Both showed 95% of similarity with each other and were 97% similar to B. bicornis (AF419313).The results suggest that new genotypes or not described protozoa of the Babesia and Hepatozoon genus are present in the study area and small mammals may be acting as reservoirs for these agents in the Pantanal region.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A Floresta Tropical Atlântica apresenta uma enorme biodiversidade, e está atualmente sujeita a inúmeras pressões como a perda de área pela intensa ocupação humana, agricultura, pecuária, urbanização e industrialização. Esses impactos têm provocado desmatamento e fragmentação florestal, processos que interferem na manutenção das populações animais, inclusive afetando os ciclos silvestres de parasitas e microorganismos. Didelphis aurita é um marsupial da Mata Atlântica com alta capacidade adaptativa a ambientes perturbados. Esta espécie onívora é tolerante à fragmentação florestal, podendo sobreviver em ambientes silvestres, rurais, suburbanos e urbanos, tendo importância na conexão dos ciclos silvestres e urbanos de diversos agentes. Este trabalho teve por objetivo descrever aspectos hematológicos, bioquímicos e de hemoparasitas em Didelphis aurita de duas áreas da Serra dos Órgãos/ RJ, uma área fragmentada e outra de mata contínua. Entre julho de 2011 e fevereiro de 2012 foram capturados 61 animais que tiveram amostras de sangue avaliadas. Os resultados expressos como média desvio padrão foram: Volume Globular 38,66 % ( 4,97); Hemácias 5,40 ( 0,75) x106/mm3; Hemoglobina 12,78 ( 1,68) g/dL; VGM 71,69 ( 3,56) fl; CHGM 33,01 ( 0,63) %; Plaquetas 514,70 ( 323,10) x 103/mm3; Leucócitos 19.678,52 ( 10.152,26)/mm3; Basófilos 0,59 ( 0,72) %; Eosinófilos 13,79 ( 6,94)%; Bastonetes 0,77 ( 2,04) %; Segmentados 41,12 ( 13,95) %; Linfócitos 41,97 ( 12,97) %; Monócitos 1,75 ( 1,51)%. Para parâmetros bioquímicos encontramos os seguintes resultados: Proteínas totais 8,50 ( 1,68); albumina 3,03 ( 0,69); globulina 5,44 ( 1,66); uréia 83,57 ( 20,11); creatinina 0,44 ( 0,13); ALT 85,01 ( 65,65); AST 314,55 ( 130,58); FA 420,38 ( 371,89); GGT 19,40 ( 8,51). Os parâmetros hematócrito, hemoglobina, hematimetria, ALT, AST e FA foram maiores nos machos do que nas fêmeas. Adultos apresentaram valores de proteína plasmática total, leucócitos, hematócrito, hemoglobina, hematimetria, albumina, proteínas totais, creatinina e GGT maiores do que jovens, e o inverso ocorreu para plaquetas, globulina e FA. Animais do Fragmento apresentaram valores de massa corporal e albumina menores do que os do Garrafão, e o inverso ocorreu para GGT e globulina. Babesiasp. ocorreu em 26,6% da população, sendo mais freqüente em adultos. Estes resultados são os primeiros parâmetros de referência para Didelphis aurita na Serra dos Órgãos, contribuindo para o estudo desta espécie. / The Atlantic Forest is characterized by a high species diversity and endemism. This ecosystem is under human-induced threats for decades, loosing area to agriculture, pastures and urban areas.These threats have lead to habitat destruction and fragmentation, disturbing ecological processes and affecting the enzootic cycle of diseases. Didelphis aurita is an Atlantic Forest marsupial with synanthropic habit, capable of survive in disturbed and fragmented environments, including urban areas. The aim of this study was to describe haemoparasites, hematologic and serum chemistry aspects of D. aurita in Serra dos Órgãos/ RJ, comparing two areas, a conserved forest and a fragmented landscape. Between July 2011 and February 2012 61 specimens were studied. Results are given as mean standard deviation, as follows: Packed cell volume 38,66 % ( 4,97); erythrocytes 5,40 ( 0,75)x106/mm3; hemoglobin 12,78 ( 1,68) g/dL; MCV 71,69 ( 3,56) fl; MCHC 33,01 ( 0,63) %; Platelets 514,70 ( 323,10) x 103/mm3; Leukocytes 19.678,52 ( 10.152,26)/mm3; basophils 0,59 ( 0,72) %; eosinophils 13,79 ( 6,94)%; band neutrophils 0,77 ( 2,04) %; neutrophils 41,12 ( 13,95) %; lymphocytes 41,97 ( 12,97) %; monocytes 1,75 ( 1,51)%. The serum chemistry values were: total protein8,50 ( 1,68); albumin 3,03 ( 0,69); globulin 5,44 ( 1,66); BUN 83,57 ( 20,11); creatinine 0,44 ( 0,13); ALT 85,01 ( 65,65); AST 314,55 ( 130,58); AP 420,38 ( 371,89); GGT 19,40 ( 8,51). Packed cell volume, hemoglobin, erythrocytes, ALT, AST and AP were higher in males than in females. Adults have higher values of total plasmatic protein, leukocytes, packed cell volume, hemoglobin, erythrocytes, albumin, total protein and GGT than young animals. The Young have higher platelets, globulin and AP. Animals from the fragmented area present lower body mass and lower levels of albumin than those from the conserved Forest. The opposite occurred for GGT and globulin. Babesia sp. were found in 26.6% of specimens, more frequently in adults. These results are the very first reportof hematologic and serum chemistry values for D. aurita in Serra dos Órgãos.

No período de julho de 2004 a junho de 2012, foram realizadas 104 capturas de lobos-guará. Os animais foram contidos quimicamente para coleta de material biológico. Carrapatos foram encontrados em 94 lobos e enviados ao Laboratório de Doenças Parasitárias da FMVZ-USP, onde foram identificados com auxílio de estereomicroscópio e chaves taxonômicas. Das amostras analisadas, foram encontradas 72 larvas, 188 ninfas e 911 carrapatos adultos, pertencetes as espécies Rhipicephalus microplus, Amblyomma spp., Amblyomma cajennense, Amblyomma ovale, Amblyomma brasiliense, Amblyomma tigrinum. O presente trabalho registra pela primeira vez larva de R. microplus e adulto de A. brasiliense parasitando C. brachyurus no país, reforçando os achados prévios da literatura destas espécies de carrapatos utilizarem os lobos-guará como hospedeiros. Sangue e soro também foram coletados durante contenção química, 67 amostras de sangue total de lobos guará e 52 carrapatos adultos foram testados através da técnica de Reação em Cadeia de Polimerase (PCR) para a pesquisa de DNA de Anaplasma spp., Babesia spp., Ehrlichia spp., Hepatozoon spp. e Rickettsia spp. Foi realizada a sorologia para a pesquisa de anticorpos anti-Rickettsia spp. para 210 amostras de soro de cães domésticos e 43 foram positivas. Quatro indivíduos apresentaram reação homóloga para R. parkeri, dois para R. rhipicephali, um para R. rickettsii. Alem disso, foram testadas 88 amostras de soro de lobo guará e, destas, 84 foram positivas para pelo menos uma das espécies de Rickettsia e indicaram reação homóloga para R. parkeri e R. rhipicephali. Foram testadas 84 amostras de soro de lobo-guará na sorologia para Ehrlichia canis e, destas, 16 foram positivas. Nos testes moleculares foi detectada e confirmada a presença de H. canis, H. felis, R. parkeri já descritas para os hospedeiros vertebrados e invertebrados testados no estudo e ainda o primeiro registro em A. tigrinum de Candidatus R. andeanae e Candidatus Midichloria mitocondrii / From July 2004 to June 2012, 104 maned wolves were captured. The animals were chemically restrained in order to collect biological material. Ticks were found in 94 wolves and sent to the Laboratory of Parasitic Diseases FMVZ USP, and were identified using a stereomicroscope and taxonomic keys. Of the samples analyzed, 72 were larvae, 188 nymphs and 911 adult ticks, from the species Rhipicephalus microplus , Amblyomma spp , A. cajennense, Amblyomma ovale, Amblyomma brasiliense e Amblyomma tigrinum. This paper reports for the first time larvae of R. b. microplus and adult of A. brasiliense parasitizing C. brachyurus in the country, reinforcing the findings in previous studies of these species of ticks using the maned wolves as hosts . Blood and serum samples were also collected during chemical restraint, 67 blood samples of maned wolves and 52 adult ticks were tested using the technique of Polymerase Chain Reaction (PCR) for the detection of DNA of Anaplasma spp., Babesia spp. Ehrlichia spp., Hepatozoon spp. and Rickettsia spp. Serology was performed for the detection of antibodies to Rickettsia spp. 210 serum samples of domestic dogs were tested and 43 were positive. Four individuals showed reaction homologous to R. parkeri, two for R. rhipicephali and one for R. rickettsia. In addition, we tested 88 serum samples of maned wolf and 84 were positive for at least one species of Rickettsia and indicated homologous reaction to R. parkeri and R. rhipicephali. 84 maned wolf serum samples were in serology for Ehrlichia canis and 16 were positive. In molecular tests was detected and confirmed the presence of H. canis, H. felis and R. parkeri had already been described for the vertebrate and invertebrate hosts tested in the study. This is also the first report of Candidatus R. andeanae and Candidatus Midichloria mitocondrii in A. tigrinum

Feline babesiosis, first described in domestic cats in South Africa in 1937, is regarded to be of great importance in the coastal regions although isolated cases also occur on the eastern highlands of Mpumalanga Province. Babesia felis (described from domestic cats) and B. leo (described from lions) are the two best characterised Babesia species in felids. These two parasites are morphologically similar when examined under a light microscope, but are serologically and genetically distinct. In this study the prevalence of these two Babesia species in various wild and domestic felid species was determined. A total of 358 samples were tested using the reverse line blot hybridization (RLB) assay. This assay makes it possible to simultaneously detect and differentiate between blood parasites using DNA probes. The RLB consists of three basic steps, the first being amplification of the variable region (V4) in the 18S rRNA gene using genus-specific primers where one is labelled with biotin. This is followed by a blotting step, where the amplicons are hybridized to oligonucleotides bound to a nitrocellulose membrane. The third and last step is the detection of the hybridized amplicons by using chemiluminescence reagents. This assay is a screening tool utilizing the variable (V4) region in the 18S rRNA gene to detect and differentiate between blood parasites. A new B. felis-specific DNA probe was developed to use in the RLB assay. Results demonstrated that these two parasites not only occur in the felid species from which they have been described, but also in other felid species. Babesia microti was also detected in various felid species, while B. rossi was detected in 1 of the lion samples. Two hundred and twelve samples tested positive for Babesia spp., of which only 54.24% of the samples reacted with the genus-specific probe. This indicates the presence of a novel Babesia or Theileria species or variant of a species. / Dissertation (MSc)--University of Pretoria, 2010. / Veterinary Tropical Diseases / unrestricted

Orientador: Henrique Nunes de Oliveira / Coorientador: Márcia Cristina de Sena Oliveira / Banca: Simone Cristina Méo Niciura / Banca: Rodrigo Giglioti / Resumo: A bovinocultura é um setor com importante destaque no agronegócio brasileiro. O carrapato Ripicephalus (Boophilus) microplus é responsável por perdas econômicas significativas aos pecuaristas e é vetor de hemoparasitoses como Anaplasma spp e Babesia spp. Sabe-se que os bovinos Bos taurus taurus são mais susceptíveis à infestação por carrapatos do que Bos taurus indicus. Acredita-se que o mesmo ocorra para a infecção por Babesia bovis. Neste trabalho, foram avaliados, em duas colheitas, 355 bovinos da raça Angus, pertencentes a uma fazenda de Uruguaiana-RS, nos quais foram realizadas contagens de carrapatos e colheitas de amostras de sangue para quantificação de B. bovis, pela técnica de qPCR, e genotipagem com chip de 150.000 marcadores SNP. Para qPCR utilizaram-se sequências iniciadoras que flanqueiam um fragmento do gene do citocromo B (mt-cytB), como oligonucleotídeos iniciadores. Após genotipagem dos bovinos com o chip Gene Seek Genomic Profiler™ (GGP-HD) da Illumina Infinium®, foi realizado imputação de genótipos, para recuperação de genótipos faltantes, e controle de qualidade. Foi realizada análise de associação genômica ampla (GWAS), para cada uma das características, infecção por B. bovis e resistência a carrapatos, através do método denominado "Single Step Genomic BLUP" (ssGBLUP). Todos os animais apresentaram infestação por carrapatos e infecção por B. bovis, determinada pela qPCR, e altos valores médios para ambas as características. Algumas regiões cromossômicas ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The cattle industry is a sector with importance in the Brazilian agribusiness. The Ripicephalus (Boophilus) microplus is responsible for economic losses and is a vector for hemoparasitoses, such as Anaplasma spp and Babesia spp. It is known that the Bos tauros animals are more susceptible to infestation by ticks when compared with infestation in Bos indicus animals. It is believed that the same behavior keeps for infection by Babesia bovis. They were evaluated, in two collections, 355 Angus cattle, from a farm in Uruguaiana city, estate of Rio Grande do Sul, where were performed tick counts, quantification of B. bovis by qPCR and genotyping with a 150K chip. Were used as primers, in the qPCR, sequences that flanking the fragment of the cytochrome b gene. The technique was standardized and optimized using specimens of isolates of B. bovis. After genotyping, imputation was carried out, for recovery of missing genotypes, and quality control. Genome association analysis was performed (GWAS), to each of the characteristics, through the method called "Single Step Genomic BLUP" (ssGBLUP). All animals showed tick infestation and infection by B. bovis and high average values for both characteristics. Some regions on chromosomes were identified as significant to the characteristics tick infestation and infection by B. bovis, and seven chromosomes, identified in the present study, were already described in other studies. The present study agrees with other results indicating that the qP... (Complete abstract click electronic access below) / Mestre

Orientador: Wilma Aparecida Starke Buzetti / Banca: Solange Maria Gennari / Banca: Rosangela Zacarias Machado / Banca: Maria Conceição Zocoller Seno / Banca: Gilson Pereira de Oliveira / Resumo: O objetivo do presente estudo foi analisar a resposta imune humoral, pelo monitoramento dos anticorpos anti-Babesia bovis, anti-Babesia bigemina e anti-Anaplasma marginale, em búfalos (Bubalus bubalis) naturalmente infectados. Para esta pesquisa, utilizaram-se amostras de soro e de colostro/leite de búfalas adultas do periparto aos 11 meses após, e de soros dos seus bezerros, durante o primeiro ano de vida nos anos de 1999/2000 e 2005. Para determinar o perfil da resposta imune humoral destes animais, utilizou-se o método ELISA indireto e os dados foram apresentados e analisados como a média de um grupo de animais, em diferentes faixas etárias e, individualmente. Após a leitura e interpretação dos dados, os resultados dos animais analisados em grupos apresentaram baixa concentração de anticorpos, ou seja, abaixo do ponto de corte (D.O. = 0,265 e NE=3) de anticorpos anti-A. marginale nos soros, durante os primeiros 90 e 105 dias após o parto e nascimento, respectivamente, para búfalas e seus bezerros. Em seguida, a concentração de anticorpos anti-A. marginale no soro dos bezerros búfalos aumentou ligeiramente acima do ponto de corte e manteve-se assim até atingirem aproximadamente um ano de idade, indicando uma imunidade adquirida, após o contato com a bactéria. Nas búfalas ocorreu soroconversão (NE acima de 3) para Babesia de ambas as espécies, por quase todo o período analisado, com uma elevação acentuada (NE=4 a NE=6) entre os dias 91 e 335 dias após o parto, fato não verificado para os bezerros, no mesmo período. No colostro/leite, os anticorpos anti-B. bovis e anti-B. bigemina foram detectados nos primeiros sete dias pós-parto, mas não foram observados no teste anti-A. marginale. Quando os animais foram analisados individualmente (duas búfalas e seus bezerros), observou-se em um dos bezerros, uma forte imunidade humoral... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim of the present study was to analyze the humoral-immune response of water buffaloes (Bubalus bubalis) naturally infected with Babesia bovis, B. bigemina and Anaplasma marginale. For this work, colostrums/milk and blood samples were weekly, fortnightly and monthly harvested prior and after partum (buffalo cows) and from birth to 365 days after birth (buffalo calves). The antibodies in the colostrums/milk and serum samples from these animals were determined using an ELISA indirect method and the data were analyzed as a mean of a group of animals with matched ages during the period of 1999/2000 or individually during the year of 2005. The data from animals analyzed in group showed that the antibodies against A. marginale were in low concentration (below the cut off point, D.O. = 0.265 and ELISA levels, EL = 3), in the sera of buffalo, during the first 90 and 105 days, respectively for cows and calves. Then, the concentration of anti-A. marginale in the serum samples of buffalo calves, slightly raised to above the cut off point and kept in higher levels up to approximately 365 days after birth, indicating acquired immunity. Serum conversion for Babesia occurred in high levels and above the cut off point only for buffalo cows for all period of experimentation. The antibody levels against Babesia for both species and Anaplasma increased in the sera of buffalo cows between the days 91 and 335 after partum. In the colostrums, anti-B. bovis and anti-B. bigemina antibodies were detected in high levels during the first seven days after partum, but then abruptly declined to zero. Anti-A. marginale, on the other hand were not detected in the colostrums of these animals. When four animals (two buffalo cows and their calves) were individually analyzed it was observed an individual variation in the immune response: in one buffalo calf there was a strong passive... (Complete abstract click electronic access below) / Doutor

Babesiosis is a disease caused by parasites of Babesia species that is spread through ticks. Babesiosis can affect humans and many other mammals all over the world. In the United States, babesiosis is mainly caused by Babesia microti with additional species of Babesia infecting dogs. Dogs have long been known to be a good indicator species for human tick-borne infection due to the shared spaces they have with humans and their tendency to explore and pick up diseases in the environment. This study used a group of hunting dogs to determine the presence of Babesia infection. The goal was to determine a predictive model for human infection. Infection was defined as having a positive antibody test or molecular test for Babesia species in blood samples. Blood samples were taken at two time points, tested, and compared. Statistical methods were used to analyze the results of the tests and compare them with variables such as region, gender, age, and other diseases the dogs were exposed to. Of 214 dogs, 56 had Babesia infections, with a prevalence rate of 26.2%. At the first time point the model showed age and infection with Anaplasma platys as being significant. At the 2nd time point, 29 dogs were lost to follow up, leaving a total of 185 dogs sampled with 63 testing positive for Babesia spp. The rate for the 2nd time point was 34.1%. No variables were found to be significant in the model for the 2nd time point. Despite differences in Babesia that infect humans and dogs, the hunting dog cohort reflects a model that validates the environmental exposures, coinfections, and demographic variables that affect transmission of the pathogen. Furthermore, findings of this study cast doubt on the ability of Ixodes scapularis to act as capable vector for canine-infecting Babesia species.

Babesia odocoilei is an intraerythrocytic apicomplexan parasite which infects cervidae, sometimes causing babesiosis. It is vectored by the tick Ixodes scapularis and is distributed throughout the southeastern United States. The geographic and host range continue to extend as new incidence of infection is detected. A genomic DNA region spanning the internal transcribed spacer 1 (ITS1), 5.8S rRNA gene, and ITS2 of ribosomal RNA (rRNA) from 18 B. odocoilei isolates (speciation confirmed by small subunit rRNA analysis) was amplified using the polymerase chain reaction, cloned and sequenced. The isolates originated from 6 different cervidae or bovidae hosts in various U.S. geographic areas. Included in the analysis was a previously described reindeer B. odocoilei-like isolate, RD61, which showed only 99.0% identity in SSU rRNA analysis to B. odocoilei. Percent identity pairwise comparisons among the samples were calculated for both the full ITS1-5.8S-ITS2 and individual genomic regions. Identity values for all comparisons ranged from 90% to 100%, with the exception of RD61, which showed no higher than 88% identity for all gene regions. An analysis of fixed differences identified in the ITS1 and ITS2 gene regions of all clones revealed 21 fixed differences in ITS1, and only 11 in ITS2. Most isolates were found to have 2 overall patterns of fixed differences, although some had 1 or 3. Phylogenetic analysis of all sequences for the entire ITS1-5.8S-ITS2 gene region placed most isolates into 2 distinct groups corresponding to those observed in the analysis of fixed differences. This suggested the presence of at least 2 rRNA transcription units in B. odocoilei. ITS analysis failed to demonstrate host or geographic differences that might serve to pinpoint the source of outbreaks of B. odocoilei in farmed and managed host animals. This failure might result from genetic recombination of ITS genomic regions during the tick vector stage. Lack of conspecificity between the RD61 isolate and B. odocoilei was supported by this study; however, more data are needed to clarify the taxonomic status of this B. odocoilei-like isolate.

Apicomplexan parasites are responsible for some of the most devastating human and veterinarian diseases and are parasites of great economic importance. Apicomplexa include Plasmodium, Toxoplasma and Babesia species. The pathogenic mechanisms developed by Apicomplexa parasites, in particular those that reside in a parasitophorous vacuole, involve considerable changes to the host cell, including the expression of variable surface proteins required for immune evasion. In Plasmodium falciparum infections, host cell remodeling is responsible for disease symptomology and severity in the human host. This work represents a multi-faceted study of antigenic secretion and the role of secreted antigens in pathogenesis. We study in detail the mechanisms of antigen secretion in Apicomplexa parasites. By use of comparative genomics, we find Plasmodium Export Element (PEXEL)-like motifs in a subset of Cryptosporidium and Babesia secreted proteins. However, in Babesia the motif functions as a spherical body targeting sequence, suggesting that secretory mechanisms in Apicomplexa are adapted to the parasite's intracellular lifestyle. To elucidate the relationship and function of exported antigens, we first focused on P. falciparum to determine gene co-expression modules. We found that in vivo, export modules are composed of constitutively or variably expressed genes, the latter group associated with patient clinical phenotypes. We then focused on a novel gene family called "phist" and show, using transcriptional expression profiling, its role in P. falciparum cytoadherence. In total, we demonstrate that antigen secretion is an evolutionary mechanism in Apicomplexa parasites and that variant expression of the genes encoding these antigens may allow parasites to adapt to environmental stresses.