Made available in DSpace on 2015-03-03T11:52:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-01Bitstream added on 2015-03-03T12:07:16Z : No. of bitstreams: 1 000811584.pdf: 1144939 bytes, checksum: 938fd09c5350bf8a032088e29e2f5f49 (MD5) / O sistema de criação conjunta ovinos/bovinos, visando melhor aproveitamento da pastagem, provendo diversificação lucrativa, pode deparar com problemas graves nas propriedades rurais, tais como as parasitoses. Dentre as mais importantes, destaca-se a hemoncose. As espécies Haemonchus placei e Haemonchus contortus são comumente encontradas parasitando bovinos e ovinos, respectivamente. A infecção de bovinos por H. contortus constitui, ainda, uma incógnita na literatura, sobretudo a brasileira. Este helminto tem sido descrito parasitando principalmente ovinos e caprinos. As espécies mais citadas parasitando bovinos são H. placei e H. similis. Portanto, objetivou-se no presente estudo avaliar a infectividade de H. contortus para bovinos e comparar sua patogenicidade à de H. placei. Para tal, foram utilizados 10 bezerros, criados e mantidos sem contato com helmintos, assim distribuídos: GI - dois bezerros não inoculados (controle), GII – quatro bezerros inoculados com larvas infectantes (L3) de H. contortus e GIII – quatro bezerros infectados com larvas (L3) de H. placei. Cada bovino recebeu 10.000 L3, de acordo com o grupo experimental, em dose única. Exames coproparasitológicos (McMaster e coproculturas) foram realizados diariamente após o 60 dia da inoculação, objetivando estabelecer o período pré-patente. Foram colhidas, semanalmente, amostras de sangue para hemogramas e dosagens bioquímicas. Decorridos 42 dias pós-infecção, os 10 bovinos foram eutanasiados e necropsiados, sendo colhido “in totum” todos os exemplares de Haemonchus. Nos abomasos foram realizados estudos anátomo-histopatológicos. O período pré patente para H. contortus foi de 20 dias e para H. placei foi de 27 dias. Dentre os parâmetros hematológicos, apenas as quantificações de hemácias apresentaram diferenças significativas (P<0,05), entre os grupos. As concentrações séricas médias de proteínas totais, albumina, ureia ... / The ovine/bovine joint breeding system, which aims for a better use of pastures, providing profitable diversification, can face severe problems in rural properties, such as parasites, amongst which Haemonchus infections stand out. H. placei and H. contortus species are commonly found parasitizing cattle and sheep, respectively. Bovine infections by H. contortus still persist as an unknown possibility in literature, especially in Brazil. Such helminth has been described parasitizing sheep and goats, while the main species cited as parasites of bovines are H. placei and H. similis. Therefore, the present study aimed to evaluate infectivity of H. contortus for bovines and compare its pathogenicity to that of H. placei. To achieve this goal, 10 calves were utilized, all created and maintained free of any contact with helminths, distributed as follows: GI – two non-inoculated calves (control group); GII – four animals inoculated with infecting larvae (L3) of H. contortus; and GIII – four bovines infected with larvae (L3) of H. placei. Each inoculated calf received 10,000 L3, according to its experimental group, in a single dose. Coproparasitological exams (McMaster and coprocultures) were performed daily, starting at the 6th post-inoculation day, seeking to establish prepatent periods. Blood samples, for cell counts and biochemical dosages, were collected on a weekly basis. 42 days after infection of experimental animals, all 10 calves were euthanized and necropsied, for subsequent “in totum” collection of all Haemonchus specimens. Anatomical and histopathological examinations were performed on all abomasums. Prepatent period for H. contortus was 20 days, and 27 days for H. placei. Amongst hematological parameters, only erythrocyte counts presented significant differences (P < 0.05) between experimental groups. Average serum concentrations of total proteins, albumin, urea, creatinin, AST and GGT presented significant ...

Orientador: Jane Megid / Banca: Luis Antonio Mathias / Banca: José Paes de Almeida Nogueira Pinto / Resumo: Foi estimada a prevalência estadual da brucelose bovina em abatedouros nos diferentes níveis de inspeção sanitária no Estado de São Paulo após a implantação do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT), instituído em 2001. Para tanto, foram amostrados 1.490 bovinos em nove frigoríficos distribuídos em diferentes regiões do estado, dos quais três com inspeção federal (SIF), três com inspeção estadual (SISP) e três com inspeção municipal (SIM). Os soros foram analisados pelas provas do antígeno acidificado tamponado (AAT), e os reagentes foram confirmados pela reação de fixação de complemento (RFC). As prevalências obtidas por meio da RFC e seus respectivos intervalos de confiança, nos diferentes serviços de fiscalização, foram 0,40% [0,00% - 0,95%] (SIF), 2,00% [0,77% - 3,23%] (SISP) e 4,29% [2,67% - 6,48%] IC95% (SIM) e no estado foi estimada em 2,21% [1,47% - 2,96%]. Com os resultados obtidos, foi possível discutir a situação da brucelose bovina no Estado de São Paulo bem como o risco à saúde a que os trabalhadores de abatedouros estão expostos e a diferença entre os sistemas de inspeção vigentes no país. Concluiu-se que a soroprevalência da brucelose bovina no Estado de São Paulo permaneceu estável. Sugere-se que a adoção de novas medidas para complementar o PNCEBT poderiam acelerar o controle e a posterior erradicação da enfermidade; e que o aumento da fiscalização nos abatedouros de inspeção estadual e municipal acrescido de vigilância ativa nesses pontos poderia auxiliar no combate à doença / Abstract: This study estimated the prevalence of bovine brucellosis in state of São Paulo as well as the prevalence in slaughterhouses with different levels of sanitary inspection in the same state after the implementation of the National Program for the Control and Eradication of Brucellosis and Tuberculosis (PNCEBT) established in 2001. Were sampled 1,490 animals in nine slaughterhouses distributed in different regions of the state, three with federal inspection (FI), three with state inspection (SI), and three with municipal inspection (MI). Sera were analyzed by Rose Bengal Test (RBT) and reaction of complement fixation test (CFT). The prevalence obtained through CFT in different inspection services were, 0.40% [0.00% - 0.95%] (FI), 2.00% [0.77% - 3.23%] (SI) e 4.29% [2.67% - 6.48%] IC95% (MI) and the state prevalence was estimated at 2.21% [1.47% - 2.96%]. With the results obtained, it was possible to evaluate the situation of the bovine brucellosis in state of São Paulo as well as the risk to public health that workers are exposed in slaughterhouses and the difference between the current inspection systems in the country . It was concluded that the seroprevalence of brucellosis in the state of São Paulo remained stable, and the adoption of new measures to enhance the PNCEBT could acelerate the control and eventual eradication of the disease. In addition, we also conclude that the increased surveillance in slaughterhouses with state and municipal inspection attached to surveillance at these points could assist in combating of disease / Mestre

Made available in DSpace on 2015-10-06T13:03:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-22. Added 1 bitstream(s) on 2015-10-06T13:18:41Z : No. of bitstreams: 1 000848970.pdf: 988571 bytes, checksum: dcc8f965284a638ff0467c668813e6f1 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Bovine Herpesvirus type 5 (BoHV-5) and bovine herpesvirus type 1 (BoHV-1), members of Herpesviridae family, Alphaherpesvirinae subfamily; may replicate in different tissues such as nervous, glandular, lymphoid and parenchymal organs and are able to remain in a latent state, lodged in nerve ganglia. In Brazil clinical cases of BoHV - 5 encephalitis are the second leading cause of death due to encephalitis with determined etiology, supplanted only by rabies, while the BoHV-1 is present in an endemic form in the national herd, being responsible for a variety of clinical signs as rhinotracheitis, abortion, conjunctivitis, vulvovaginitis and balanoposthitis. It is expected that a viral agent, to infect a cell, activate the pathways of programmed cell death process, however there are reports that the gene related to latency (RL) of Herpesvirus is able to inhibit cell apoptosis, protecting the infected cell during latency through unknown mechanisms. Therefore, this study demonstrated the capacity of BoHV-5 and BoHV-1 to manipulate mitochondrial metabolism of the infected cell, interfering with intrinsic pathway of apoptosis

Made available in DSpace on 2015-12-10T14:23:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-13. Added 1 bitstream(s) on 2015-12-10T14:30:01Z : No. of bitstreams: 1 000851407.pdf: 349620 bytes, checksum: c4f28202679e08ad9ffcf30801ef536a (MD5) / Foi estimada a prevalência estadual da brucelose bovina em abatedouros nos diferentes níveis de inspeção sanitária no Estado de São Paulo após a implantação do Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT), instituído em 2001. Para tanto, foram amostrados 1.490 bovinos em nove frigoríficos distribuídos em diferentes regiões do estado, dos quais três com inspeção federal (SIF), três com inspeção estadual (SISP) e três com inspeção municipal (SIM). Os soros foram analisados pelas provas do antígeno acidificado tamponado (AAT), e os reagentes foram confirmados pela reação de fixação de complemento (RFC). As prevalências obtidas por meio da RFC e seus respectivos intervalos de confiança, nos diferentes serviços de fiscalização, foram 0,40% [0,00% - 0,95%] (SIF), 2,00% [0,77% - 3,23%] (SISP) e 4,29% [2,67% - 6,48%] IC95% (SIM) e no estado foi estimada em 2,21% [1,47% - 2,96%]. Com os resultados obtidos, foi possível discutir a situação da brucelose bovina no Estado de São Paulo bem como o risco à saúde a que os trabalhadores de abatedouros estão expostos e a diferença entre os sistemas de inspeção vigentes no país. Concluiu-se que a soroprevalência da brucelose bovina no Estado de São Paulo permaneceu estável. Sugere-se que a adoção de novas medidas para complementar o PNCEBT poderiam acelerar o controle e a posterior erradicação da enfermidade; e que o aumento da fiscalização nos abatedouros de inspeção estadual e municipal acrescido de vigilância ativa nesses pontos poderia auxiliar no combate à doença / This study estimated the prevalence of bovine brucellosis in state of São Paulo as well as the prevalence in slaughterhouses with different levels of sanitary inspection in the same state after the implementation of the National Program for the Control and Eradication of Brucellosis and Tuberculosis (PNCEBT) established in 2001. Were sampled 1,490 animals in nine slaughterhouses distributed in different regions of the state, three with federal inspection (FI), three with state inspection (SI), and three with municipal inspection (MI). Sera were analyzed by Rose Bengal Test (RBT) and reaction of complement fixation test (CFT). The prevalence obtained through CFT in different inspection services were, 0.40% [0.00% - 0.95%] (FI), 2.00% [0.77% - 3.23%] (SI) e 4.29% [2.67% - 6.48%] IC95% (MI) and the state prevalence was estimated at 2.21% [1.47% - 2.96%]. With the results obtained, it was possible to evaluate the situation of the bovine brucellosis in state of São Paulo as well as the risk to public health that workers are exposed in slaughterhouses and the difference between the current inspection systems in the country . It was concluded that the seroprevalence of brucellosis in the state of São Paulo remained stable, and the adoption of new measures to enhance the PNCEBT could acelerate the control and eventual eradication of the disease. In addition, we also conclude that the increased surveillance in slaughterhouses with state and municipal inspection attached to surveillance at these points could assist in combating of disease

Made available in DSpace on 2014-06-11T19:32:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-04-17Bitstream added on 2014-06-13T19:22:37Z : No. of bitstreams: 1 lima_rca_dr_jabo.pdf: 689333 bytes, checksum: b8de9b1893e535c64bce7067077a1c6b (MD5) / A criptosporidiose é uma enfermidade de veiculação hídrica, possui como agravante a dificuldade de prevenção da contaminação ambiental e ausência de medidas terapêuticas eficazes. Com acentuada importância na bovinocultura, ocasiona inflamação e atrofia das vilosidades intestinais resultando em perda da superfície de absorção. O presente estudo teve como objetivo caracterizar molecularmente a infecção por Cryptosporidium spp. em bezerros do Município de Formiga, região Centroeste de Minas Gerais. Um total de 300 amostras de fezes de bezerros holandeses, nelore e sem raça definida, foram avaliadas pela técnica de coloração contraste negativo de verde malaquita e por meio da reação de nested-PCR para amplificação de fragmentos de DNA da subunidade 18S do gene do RNA ribossômico. Ocorrência de 5,33% (16/300) pelo verde malaquita e 4,66% (14/300) pela PCR foram observadas, sendo que por meio da caracterização molecular foram identificadas as espécies Cryptosporidium andersoni e Cryptosporidium ryanae. Os bezerros infectados apresentavam baixa eliminação de oocistos de Cryptosporidium spp. Não houve sendo que não houve relação da positividade com todas as variáveis estudadas. No sequenciamento foi encontrado as espécies de C. andersoni e C. ryanae, inclusive parasitando bezerros em faixas etárias, não relatadas na literatura. Estas duas espécies de Cryptosporidium, caracterizados molecularmente (Nested-PCR), no presente estudo, são descritas pela primeira vez parasitando bovinos do estado de Minas Gerais, Brasil / Cryptosporidiosis is a waterborne disease, has as aggravating the difficulty of preventing environmental contamination and the absence of effective therapeutic measures. With pronounced importance to the cattle, causes villous atrophy and inflammation resulting in loss of intestinal absorption surface. The present study aimed to characterize molecularly infection by Cryptosporidium spp. calves in the City of Formiga, Midwest region of Minas Gerais. A total of 300 stool samples from Holstein calves, and Nelore breed were evaluated by negative contrast staining of malachite green and through the reaction of nested-PCR for amplification of DNA fragments of the 18S subunit of the RNA gene ribosomal. Occurrence of 5.33% (16/300) for malachite green and 4.66% (14/300) were observed by PCR, and by molecular characterization were identified species Cryptosporidium andersoni and Cryptosporidium ryanae. Calves infected had lower elimination of Cryptosporidium spp. There was no positive relationship with all the variables. Sequencing were found in the species C. andersoni and C. ryanae, including parasitizing calves in age, not reported in the literature. These two species of Cryptosporidium, molecularly characterized (nested PCR), the present study describes for the first time parasitizing cattle in the state of Minas Gerais, Brazil

Made available in DSpace on 2015-04-09T12:28:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-27Bitstream added on 2015-04-09T12:48:04Z : No. of bitstreams: 1 000814098.pdf: 1490588 bytes, checksum: ac5274fb5b2345e0c2260a454c1dfdd0 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Bovine Herpesvirus type 5 (BoHV-5) infection results in non- suppurative meningoencephalitis which underlies significant economic losses in South America. Mesenchymal stem cells (MSCs) are found in fetal tissues, such as the umbilical cord. The goal of this research was to characterize the isolation and viability of bovine umbilical cord MSCs for in vitro differentiation into neuron-like cells (NLC) to be used as a model for experimental infection with BoHV-5. MSCs were isolated from Wharton's jelly and differentiated in vitro into adipogenic, chondrogenic, and osteogenic cells as well as NLC. The cells lines were characterized molecularly and immunophenotypically, using PCR, immunocytochemistry, and flow cytometry. The set of primers and panel of primary antibody used in the experiments confirmed the isolation of mesenchymal stem cells and their differentiation into neuronal-like cells. After adaptation and differentiation, the cells were infected with BoHV-5, exhibiting typical cytopathic effect and increased viral replication rate 72 h post-infection. Viral particles were identified in the NLC and in the extracellular environment. by in situ hybridization These results corroborate the use of MSCs as a source of NLC which can be used to study the pathogenesis of the experimental in vitro infection wtih BoHV-5 / FAPESP: 2010/50626-5

Made available in DSpace on 2015-08-20T17:10:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-03-26. Added 1 bitstream(s) on 2015-08-20T17:25:54Z : No. of bitstreams: 1 000833070.pdf: 276509 bytes, checksum: 168d0b0f9090ec30681d2c2cccd8a266 (MD5) / Este experimento analisou os efeitos da administração do meloxicam nas respostas fisiológicas e no desempenho de bovinos na fase inicial do confinamento após serem transportados. Foram utilizados 84 bezerros Angus × Hereford, divididos por peso vivo (PV) no dia -10 do experimento e distribuídos em 21 baias de confinamento. Do d -10 ao d 0, todos os animais receberam uma dieta composta por feno de alfalfa a vontade e 2,4 kg/animal (base MS) de um concentrado a base de milho. No dia 0, todas as baias foram aleatoriamente atribuídas a receberem um dos seguintes tratamentos: 1) transporte por 1.440 km em um caminhão comercial e administração oral de meloxicam (1 mg/kg do PV) no carregamento (d 0), no descarregamento (d 1) e diariamente do d 2 ao d 7 no período inicial de confinamento (MEL; n = 7), 2) transporte e administração do tratamento nos mesmos dias do grupo MEL, porém com lactose monohidratada (1 mg/kg do PV) (TRANS; n = 7), 3) ausência de transporte e administração de lactose monohidratada (1 mg/kg do PV) nos mesmos dias dos tratamentos MEL e TRANS (CON; n = 7). Logo após o descarregamento (d 1), todos os animais dos tratamentos MEL e TRANS retornaram para suas respectivas baias para o período de 21 dias de confinamento, recebendo a mesma dieta oferecida do d -10 ao d 0. Nos dias 0 e 1, os tratamentos foram aplicados através de solução aquosa e administrados via oral, porém dos dias 2 ao 7, os tratamentos foram misturados no concentrado. Para o cálculo de ganho peso diário (GPD) foram utilizados as médias de ... / This experiment evaluated the effects of meloxicam administration on physiological and performance responses of transported cattle during feedlot receiving. Eighty-four Angus × Hereford steers were ranked by BW on d -10, and assigned to 21 dry lot pens. From d -10 to 0, pens were fed Alfalfa-grass hay ad libitum and 2.4 kg/steer daily (DM basis) of a corn-based concentrate. On d 0, pens were randomly assigned to: 1) transport for 1,440 km in a livestock trailer and oral administration of meloxicam (1 mg/kg of BW) at loading (d 0), unloading (d 1), and daily from d 2 to 7 of feedlot receiving (MEL; n = 7), 2) same transportation and treatment schedule of MEL, but oral administration of lactose monohydrate (1 mg/kg of BW) instead of meloxicam (TRANS; n = 7), or 3) no transport and oral administration of lactose monohydrate (1 mg/kg of BW) concurrently with treatment administration to MEL and TRANS (CON; n = 7). Upon arrival (d 1), MEL and TRANS steers returned to their pens for a 21-d feedlot receiving with the same diet offered from d -10 to 0. Treatments were administered to steers via oral drench on d 0 and 1, or mixed daily with the concentrate from d 2 to 7. Full BW was recorded prior to (d -2, -1, and 0) treatment application and at the end of experiment (d 20, 21, and 22) for ADG calculation. Daily DMI was recorded from d 1 to 21. Blood samples were collected on d 0, 1, 3, 5, 7, 10, 14, and 21. During the initial 7 d of feedlot receiving, hay and total DMI were reduced (P ≤ 0.03) in TRANS vs. CON and MEL, similar between CON and MEL (P ≥ 0.26), whereas concentrate DMI did not differ (P = 0.16) among ...

Made available in DSpace on 2014-12-02T11:16:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-28Bitstream added on 2014-12-02T11:21:18Z : No. of bitstreams: 1 000784203.pdf: 2707587 bytes, checksum: 60463c95eda02952a5fde820ad4e0531 (MD5). Added 1 bitstream(s) on 2015-11-19T17:38:36Z : No. of bitstreams: 1 000784203_20160228.pdf: 79467 bytes, checksum: 21171e215aeff9c2ee4e0d2294237064 (MD5) Bitstreams deleted on 2016-02-29T12:16:28Z: 000784203_20160228.pdf,. Added 1 bitstream(s) on 2016-02-29T12:17:26Z : No. of bitstreams: 1 000784203.pdf: 2707582 bytes, checksum: 3f7192c7784f1822d5d754fa4604956e (MD5). Added 1 bitstream(s) on 2016-03-07T17:59:40Z : No. of bitstreams: 1 000784203_20180228.pdf: 109002 bytes, checksum: 424f5f1795170ba23918fbed01d7890b (MD5) Bitstreams deleted on 2018-03-02T12:40:00Z: 000784203_20180228.pdf,. Added 1 bitstream(s) on 2018-03-02T12:40:57Z : No. of bitstreams: 1 000784203.pdf: 2707585 bytes, checksum: 744cb15abdfa0446e7f2731acecf8660 (MD5) / Infecções por nematódeos gastrintestinais causam consideráveis perdas na pecuária. Haemonchus spp. é o principal nematódeo que acometem bovinos e ovinos principalmente em regiões tropicais e subtropicais. Portanto este estudo teve por objetivo avaliar metodologias de diagnóstico baseado em analises morfológicas e moleculares de larvas infectantes e adultos de Haemonchus spp. Foram utilizados 23 bezerros e 20 cordeiros naturalmente infectados por nematódeos gastrintestinais. Antes da necropsia dos animais, fezes foram coletadas para realização de coproculturas e obtenção de larvas infectantes, que foram morfometricamente analisadas. Logo após o sacrifício dos animais, os abomasos foram removidos e congelados, para posterior recuperação dos exemplares de Haemonchus spp. que foram armazenados em álcool 70º. No caso dos ovinos, os parasitas permaneceram estocados em alcool 70º por mais de um ano antes de serem analisados, enquanto que os parasitas dos bovinos foram armazenados por apenas quatro meses. O diagnóstico morfológico dos Haemonchus machos foi realizado por meio da análise de sínlofe e mensuração de espículos. Os mesmos parasitas utilizados na análise morfológica foram submetidos à análise molecular, com utilização da PCR, com os primers 52/154, 75/86 e Hplacbotu. A técnica padrão ouro para identificação de Haemonchus similis foi análise de espículos e para Haemonchus placei foi PCR com os primes HplacBotu. Todas as amostras de ovinos foram consideradas como sendo de Haemonchus contortus, devido à especificidade parasitária e nenhuma evidencia de outras espécies de Haemonchus nas análises moleculares. Os resultados demonstraram diferença estatística no comprimento da distância entre o final da L3 e o final da bainha das larvas infectantes das três espécies. Houve sobreposição de medidas de apenas 1,26% entre as espécies H. placei e H. contortus. Na análises moleculares o par ... / Nematodes gastrointestinal infection causes considerable losses in the livestock industry. Haemonchus spp. is major nematode that affects cattle and sheep mainly in tropical and subtropical region. Therefore, the aim of the present study was to assess diagnosis methodologies based in morphological and molecular analysis of infective larvae and adults of Haemonchus spp. Twenty-three calves and twenty lambs naturally infected with gastrointestinal nematodes were utilized. Before necropsy of animals, faecal samples were collated for preparing faecal cultures and obtaining infective larvae that were morphometrically analyzed. After the euthanasia of animals the abomasums were removed and frozen for posterior recovered Haemonchus specimens that were stored in alcohol 70º. In the case sheep, the parasites remained stored in alcohol 70º for more than one year before be analyzed, while the parasites of cattle were stored for only four months. The morphological diagnosis of males Haemonchus spp. was released by synlophe analysis and spicule measurements. The same parasites used in the morphological analysis were submitted in the molecular analysis, using PCR, with primers 52/159, 75/86 and HplacBotu. The “gold standard” method used to identify Haemonchus similis was spicules analysis and to identify Haemonchus placei was PCR with the primers HplacBotu. All Haemonchus specimens recovered from sheep were assumed to be Haemonchus contortus based on host specificity, and because the molecular analysis did not show any evidence of other Haemonchus species. The results presented statistic differences in the sheath tail length of infective larvae of three species. There was overlapping of measurements was only 1.26% between H.contortus and H. placei. In the espicule analysis also were observed same overlapping of measurements that caused misidentification of H. placei and H. contortus. In relation the synlophe most samples ...

Made available in DSpace on 2014-06-11T19:27:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-08-09Bitstream added on 2014-06-13T18:31:12Z : No. of bitstreams: 1 aquino_mcc_me_araca.pdf: 684677 bytes, checksum: 7318d697f23b509c8008184e6a67c272 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / om o objetivo de determinar a ocorrência e caracterizar molecularmente a infecção por Cryptosporidium spp. em bezerros bubalinos do Estado de São Paulo, Brasil, foram colhidas 222 amostras fecais de animais da raça Murrah, com até seis meses de idade. As amostras foram avaliadas pela técnica de Ziehl-Neelsen modificada e por meio da reação em cadeia da polimerase tipo nested (nPCR) para amplificação de fragmentos de DNA da subunidade 18S do gene do RNA ribossômico e sequenciamento dos fragmentos amplificados. Pela técnica de Ziehl-Neelsen modificada, foi detectada positividade de 8,1% (18/222), e pela nPCR, foi observada amplificação em 48,2% (107/222) das amostras, das quais 63 foram sequenciadas. A análise das sequências obtidas mostrou que a espécie mais frequente nesses animais foi Cryptosporidium ryanae, em bezerros bubalinos a partir de cinco dias de idade. A espécie zoonótica Cryptosporidium parvum foi detectada em apenas um animal e um genótipo incomum, similar a Cryptosporidium sp. W20486, foi encontrado, pela primeira vez em búfalos / With the aim of determining the occurrence and molecularly characterizing infection by Cryptosporidium spp. in buffalo calves from São Paulo State, Brazil, 222 fecal samples were collected from Murrah animals aged up to six months. The samples were assessed by means of modified Ziehl-Neelsen technique and nested polymerase chain reaction (nPCR) for amplification of DNA fragments from subunit 18S of the gene of ribosomal RNA and sequencing of the amplified fragments. The modified Ziehl-Neelsen technique indicated 8.1% positivity (18/222), while nPCR revealed amplification for 48.2% (107/222) samples, of which 63 were sequenced. The analysis of the obtained sequences showed that the most frequent species in these animals was Cryptosporidium ryanae, present in buffalo calves from five days of age. The zoonotic specie Cryptosporidium parvum was detected in only one animal, and an uncommon genotype, similar to that of Cryptosporidium sp. W20486, was found for the first time in buffaloes

Orientador: Alvimar José da Costa / Coorientador: Antônio Sérgio Ferraudo / Banca: Kátia Denise Saraiva Bresciani / Banca: Gilson Pereira de Oliveira / Banca: Claudio Alessandro Massamitsu Sakamoto / Banca: Welber Daniel Zanetti Lopes / Resumo: A criptosporidiose é uma enfermidade de veiculação hídrica, possui como agravante a dificuldade de prevenção da contaminação ambiental e ausência de medidas terapêuticas eficazes. Com acentuada importância na bovinocultura, ocasiona inflamação e atrofia das vilosidades intestinais resultando em perda da superfície de absorção. O presente estudo teve como objetivo caracterizar molecularmente a infecção por Cryptosporidium spp. em bezerros do Município de Formiga, região Centroeste de Minas Gerais. Um total de 300 amostras de fezes de bezerros holandeses, nelore e sem raça definida, foram avaliadas pela técnica de coloração contraste negativo de verde malaquita e por meio da reação de nested-PCR para amplificação de fragmentos de DNA da subunidade 18S do gene do RNA ribossômico. Ocorrência de 5,33% (16/300) pelo verde malaquita e 4,66% (14/300) pela PCR foram observadas, sendo que por meio da caracterização molecular foram identificadas as espécies Cryptosporidium andersoni e Cryptosporidium ryanae. Os bezerros infectados apresentavam baixa eliminação de oocistos de Cryptosporidium spp. Não houve sendo que não houve relação da positividade com todas as variáveis estudadas. No sequenciamento foi encontrado as espécies de C. andersoni e C. ryanae, inclusive parasitando bezerros em faixas etárias, não relatadas na literatura. Estas duas espécies de Cryptosporidium, caracterizados molecularmente (Nested-PCR), no presente estudo, são descritas pela primeira vez parasitando bovinos do estado de Minas Gerais, Brasil / Abstract: Cryptosporidiosis is a waterborne disease, has as aggravating the difficulty of preventing environmental contamination and the absence of effective therapeutic measures. With pronounced importance to the cattle, causes villous atrophy and inflammation resulting in loss of intestinal absorption surface. The present study aimed to characterize molecularly infection by Cryptosporidium spp. calves in the City of Formiga, Midwest region of Minas Gerais. A total of 300 stool samples from Holstein calves, and Nelore breed were evaluated by negative contrast staining of malachite green and through the reaction of nested-PCR for amplification of DNA fragments of the 18S subunit of the RNA gene ribosomal. Occurrence of 5.33% (16/300) for malachite green and 4.66% (14/300) were observed by PCR, and by molecular characterization were identified species Cryptosporidium andersoni and Cryptosporidium ryanae. Calves infected had lower elimination of Cryptosporidium spp. There was no positive relationship with all the variables. Sequencing were found in the species C. andersoni and C. ryanae, including parasitizing calves in age, not reported in the literature. These two species of Cryptosporidium, molecularly characterized (nested PCR), the present study describes for the first time parasitizing cattle in the state of Minas Gerais, Brazil / Doutor