Submitted by GUILHERME SICCA LOPES SAMPAIO null (guilhermeslsampaio@yahoo.com.br) on 2017-01-30T16:59:48Z No. of bitstreams: 1 Tese de Doutorado - Guilherme S. L. Sampaio (versão final).pdf: 1435860 bytes, checksum: d3afaf1a274ca2aaa24f6ebb66f4ae63 (MD5) / Rejected by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: Incluir o número do processo de financiamento nos agradecimentos da dissertação/tese. Corrija esta informação e realize uma nova submissão com o arquivo correto. Agradecemos a compreensão. on 2017-02-01T12:47:13Z (GMT) / Submitted by GUILHERME SICCA LOPES SAMPAIO null (guilhermeslsampaio@yahoo.com.br) on 2017-02-01T12:59:33Z No. of bitstreams: 1 Tese de Doutorado - Guilherme S. L. Sampaio (versão final).pdf: 1436609 bytes, checksum: a731559ab98fc75f6ee45c3b1a2e8208 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-02-01T13:09:02Z (GMT) No. of bitstreams: 1 sampaio_gsl_dr_bot.pdf: 1436609 bytes, checksum: a731559ab98fc75f6ee45c3b1a2e8208 (MD5) / Made available in DSpace on 2017-02-01T13:09:02Z (GMT). No. of bitstreams: 1 sampaio_gsl_dr_bot.pdf: 1436609 bytes, checksum: a731559ab98fc75f6ee45c3b1a2e8208 (MD5) Previous issue date: 2017-01-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Um total de 180 bovinos Bos indicus (Nelore), terminados em confinamento, foi dividido em grupos com 6, 12 ou 24 horas de jejum pré-abate para verificar se a redução deste tempo resulta em melhor condição higiênica e sanitária durante o processo de abate. Amostras de fezes pré e pós-jejum, de pele e de carcaça foram colhidas e analisadas para ocorrências e contagens de bactérias aeróbias mesófilas, Escherichia coli genérica, coliformes totais, assim como, E. coli produtora de toxinas Shiga (STEC) e enteropatogênica (EPEC). As amostras de fezes também foram analisadas quanto ao pH e os teores percentuais de matéria seca (MS%) e de fibras em detergente neutro (FDN%) e ácido (FDA%). Os dados foram analisados com modelos logísticos, análises de variância e regressões, ao nível de 5% de significância. A extensão do tempo de jejum pré-abate reduziu a MS%, mas elevou o pH, FDN% e FDA% das fezes. O gradual aumento do jejum pré-abate resultou em maior excreção fecal de STEC (stx1 e stx2), entretanto, com decréscimo de aeróbios mesófilos, mas sem interferência na E. coli genérica. A extensão do tempo de jejum pré-abate também resultou na maior contaminação da pele por STEC (stx1) e EPEC, assim como, das carcaças por aeróbios mesófilos, E. coli genérica e coliformes totais. Dessa forma, conclui-se que a condição higiênica e sanitária durante o processo de abate de bovinos pode ser melhorada com a redução do tempo de jejum pré-abate. Para as indústrias da carne isto pode representar a redução de custos operacionais e com a infraestrutura dos currais de abate, menos condenações e falhas no controle de produção, maior rendimento produtivo e incremento na validade comercial. A redução de STEC, além de ser um fomento à saúde publica, constitui-se em um estimulo à economia brasileira pela redução de barreiras comerciais e menor devolução de mercadorias. / A total of 180 Bos indicus (Nellore) cattle finished in feedlot were divided into groups of 6, 12 or 24 hours of pre-harvest fasting to verify if time reduction results in better hygiene and sanitary conditions during the slaughter process. Pre-fasting and post-fasting samples of feces, and post-fasting samples of hide, and carcass were collected and analyzed for the occurrence and counting of mesophilic aerobic bacteria, generic Escherichia coli, total coliforms, as well as Shiga toxin-producing E. coli (STEC) and enteropathogenic E. coli (EPEC). The feces samples were also analyzed for pH and content percentage of dry matter (DM%), neuter detergent fiber (NDF%) and acid detergent fiber (ADF%). Data were analyzed with logistic models, analyses of variance and regressions at 5% significance. The extension of pre-harvest fasting time reduced DM% but increased pH, NDF% and ADF% of feces. The gradual increase of pre-harvest fasting time resulted in greater fecal excretion of STEC (stx1 and stx2) with reduction of mesophilic aerobic bacteria, but without interference in generic E. coli. The extension of this time also resulted in greater contamination of hide by STEC (stx1) and EPEC as well as of carcass by mesophilic aerobic bacteria, generic E. coli and total coliforms. Thus, it was concluded that the hygiene and sanitary condition during the slaughter process of bovines can be improved with the reduction of pre-harvest fasting time. For the meat industry, can represent a reduction of costs with operations and facilities of lairage pens, less with condemnations and failures in the production control besides a greater productive yield and increment in the commercial validity. STEC reduction, besides being a development in public health, becomes a stimulus to the Brazilian economy by decreasing commercial barriers and return of goods. / FAPESP: 2015/02208-3 / FAPESP: 2014/01109-9

O objetivo deste trabalho foi desenhar primers, amplificar fragmentos de três loci que codificam antígenos de superfície de protozoários do gênero Sarcocystis spp. isolados do intestino de marsupiais do gênero Didelphis spp., sequenciar os fragmentos gênicos de todos isolados e compará-los entre si e com fragmentos de sequências homólogas disponíveis no GenBank. Trinta e duas amostras de Sarcocystis spp. de gambás do Estado do Rio Grande do Sul, Brasil, tiveram o DNA extraído e amplificado utilizando marcadores moleculares em genes codificadores de antígeno de superfície (SAG-2, SAG-3, e SAG-4). Entre essas amostras, 28 tiveram pelo menos um dos fragmentos gênicos sequenciados. Foi possível sequenciar os 3 fragmentos gênicos de 20 amostras. As análises das sequências dos genes renderam os seguintes resultados: SAG-2: 275 nucleotídeos e sete alelos para 26 amostras; SAG-3: 353 nucleotídeos e seis alelos para 21 amostras; SAG-4: 278 nucleotídeos e onze alelos para 25 amostras. Para cada marcador, análises filogenéticas foram realizadas empregando métodos de distância. As reconstruções filogenéticas permitiram verificar as relações de ancestralidade entre os diferentes alelos, que foram nomeados de acordo com o critério de evolução inferido na topologia das árvores. Para os três loci, diferentes alelos foram agrupados em três grupos ou genótipos, que foram nomeados com caracteres em números romanos I, II, III e IV. Diferenças intra-genótipo (sub-genótipos) foram representados por letras minúsculas (Ia, Ib, Ic, etc.). Cada alelo foi nomeado com numeração arábica (Ia1, Ia2, Ia3, etc.). Nas analises filogenéticas concatenadas baseada em dados de aminoácidos foi possível dividir as amostras dentro de três grupos. A filogenia baseada nas sequências de aminoácidos indica que três grupos de organismos devem existir dentro do complexo de indivíduos da população estudada (Sarcocystis-RS, Falcatula-like e Neurona-like). Embora o grupo designado como Sarcocystis-RS tenha um único alelo para o locus gênico SAG-3 (configuração do tipo III), táxons deste grupo compartilham alelos com indivíduos do grupo Falcatula-like. Assim, seria plausível assumir que exista uma troca gênica entre as duas populações. No que diz respeito ao grupo Neurona-like, nenhum dos indivíduos deste grupo compartilham alelos com indivíduos de outros grupos. No entanto, esta observação precisa ser confirmada, pois as análises foram baseadas em poucas sequências Neurona-like (duas sequências). Este relato revela uma notável diversidade genética entre os isolados de Sarcocystis spp de gambás do Brasil. / The present work aimed to design primers and amplify fragments of three loci that code for surface antigens of the protozoan genus Sarcocystis spp. isolated from intestine of marsupials of the genus Didelphis spp and to sequence the gene fragments of all isolates and compare them with each other and with fragments of homologous sequences available in GenBank. Thirty two samples of Sarcocystis spp. from opossums from the State of Rio Grande do Sul, had the nuclear DNA extracted and amplified using the molecular markers targeted to genes encoding surface antigens (SAG-2, SAG-3, and SAG-4). Among these samples, 28 had at least one of the gene fragments sequenced. It was possible to sequence the three gene fragments from 20 samples. The analysis of gene sequences yielded the following results: SAG-2: 275 nucleotides and seven alleles for 26 samples; SAG-3: 353 nucleotides and six alleles for 21 samples; SAG-4: 278 nucleotides and eleven alleles for 25 samples. For each marker phylogenetic analysis was performed employing distance methods. The phylogenetic reconstructions allowed us to verify the ancestry relationships between different alleles, which were named according to the criteria of evolution inferred from the tree topology. For the three loci, different alleles were grouped into three groups or genotypes, which were named with characters in Roman numerals I, II, III and IV. Intra-genotype differences (sub-genotypes) were represented by lowercase letters (Ia, Ib, Ic, etc.). Each allele was named with Arabic numerals (Ia1, Ia2, Ia3, etc.). With concatenated phylogenetic analysis based on amino acid dataset it was possible to divide the samples into three groups. The amino acid based phylogeny indicates that three groups of organisms must exist within the complex of individuals in the population studied (Sarcocystis-RS, Falcatula-like, and Neurona-like). Although the group designated as Sarcocystis-RS has a unique allele for the genetic locus SAG-3 (configuration type III), the taxa of this group share alleles with individuals in the Falcatula-like. Thus, it is plausible to assume that gene exchange occurs between these two populations. Regarding the Neurona-like group, none of the individuals in this group share alleles with individuals of the other groups. However, this observation remains to be confirmed, because this analysis was based on very few Neurona-like sequences (two sequences). This report reveals a striking genetic diversity among Sarcocystis spp isolated from opossums from Brazil.

Made available in DSpace on 2014-10-09T12:36:05Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:59:36Z (GMT). No. of bitstreams: 1 03641.pdf: 1524540 bytes, checksum: 9eac945221973ca315eb33b118d750af (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-07-29T15:13:50Z (GMT). No. of bitstreams: 1 tese helio ciencia animal.pdf: 455708 bytes, checksum: 78cc58c19ba93b461c2e9c3fbc8b9a4a (MD5) Previous issue date: 2009-03-27 / The objective of this study was to evaluate the effects of high grain diets on performance, carcass characteristics, feeding behavior and fecal parameters of 20 Nelore bovines with 28 months of age. The diets DT + BIN (10% sugar cane bagasse, 54.52% cracked sorghum, 10.94% whole cotton seed hulls, 18% soybean hulls, 2.54% soybean meal and 4% powder vitamin/mineral supplement), MGI (10% soybean hulls, 75% whole corn grain and 15% pellet vitamin/mineral supplement) and DT (44.44% cracked sorghum, 16.7% whole cotton seed hulls, 28.89% soybean hulls and 10% powder vitamin/mineral supplement) were offered to the animals in tie stalls in a completely randomized experimental design. Intake was determined through subtraction of the feed minus orts. Diets were offered to the animals in two daily meals, at 8h (60%) and 17h (40%), aiming approximately 5% of orts. Animals were weighed in the beginning of the experiment and every 21 days, always in water fasting and 14 hours after the last meal. The pH and the lengths of the half-carcasses were measured in the cool store after the cooling process had been completely finished. The areas of AOL and EGSO were taken from the left half-carcass in the 12th rib. Registrations of time spent with feeding, rumination, rest and water intake were done by visual observations of the animals every five minutes during 24 hours. For determination of feces pH and fecal starch, samples of feces were taken from the rectum of each bovine. There was difference for final weight and metabolic weight (P<0.05). Animals fed the diet DT + BIN (498.43 kg final weight and 93.27 kg metabolic weight) and the diet MGI (481.67 kg final weight and 90.32 kg metabolic weight) were more efficient than the ones fed the diet DT (456.43 kg final weight and 88.52 kg metabolic weight). Dry matter intake was greatest for the animals fed the diet DT+BIN (9.24 kg/day) and MGI (7.34 kg/day) than animals fed the diet DT (6.92 kg/day). When intake was expressed as percentage of bodyweight, the treatment DT + BIN was 20.99% higher than MGI and 26.59% than DT. There was difference (P<0.05) in the weight of fresh carcass (PCQ) and thickness of subcutaneous fat (EGSS). PCQ in the treatment DT + BIN (281.81 kg) was higher than DT (238.2 kg), but did not differ from MGI (268.98 kg). Likewise, EGSS in the diet DT+BIN was 12.36% higher than the diet MGI and 50% higher than the diet DT. There was difference for time spent with mastication, rumination, rest and water intake among diets (P<0.05), except for the time spent with feeding. Times spent for rumination and mastication were higher in the treatment DT + BIN (181.2 and 278.4 min/day), however there was no difference (P>0.05) between the diets MGI (94.2 and 171 min/day) and DT (60 and 148.2 min/day). Times spent for rest and water intake differed among treatments, with means of 436.8 and 4.46 min/day in the diet DT+BIN, 546 and 2.81 min/day in the diet MGI and 568.8 and 4.02 in the diet DT. Means of percentage of fecal starch, fecal dry matter, pH in the site of starch fermentation and starch intake were not influenced by diets (P>0.05). Score of fecal consistency and feces NDF were affected (P<0.05) by treatments, with means of 3.12 and 46.29% in the diet DT+BIN, 2.92 and 35.65% in the diet MGI and 3.2 and 48.97% in the diet DT. The addition of 10% of BIN in high grain diet improves dry matter intake, weight gain and carcass quality. The MGI diet was greater than the DT diet in terms of daily weight gain at 84 days and fresh carcass weight. Times spent for rumination, mastication and feed efficiency (g NDF/h) and (g peNDF/h) were greatest in the diet DT+BIN. The fecal consistency evaluated in the animals belonged to the diet DT+BIN, scored as firm, suggests that these animals presented better ruminal fermentation / O objetivo deste trabalho foi de avaliar os efeitos de dietas de alta proporção de concentrado, sobre os desempenhos e características de carcaça, comportamento ingestivo e indicadores fecais de 20 bovinos Nelore, com idade de 28 meses. As dietas DT+BIN (10% de bagaço de cana in natura , 54,52% de sorgo moído, 10,94% de caroço de algodão, 18% de casca de soja, 2,54% de farelo de soja e 4% de núcleo farelado), MGI (10% de casca de soja, 75% de milho grão e 15% de núcleo peletizado) e DT (44,44% de sorgo moído, 16,70% de caroço de algodão, 28,89% de casca de soja e 10% de núcleo farelado) foram distribuídas aos animais em delineamento inteiramente casualizado em baias individuais. O consumo das dietas e dos nutrientes, foi determinado mediante diferenças entre o oferecido e as sobras. As dietas, depois de pesadas, foram distribuídas aos animais na forma de mistura total em duas refeições diárias, sendo 60% do total às 8h e 40% do total às 17h, permitindo-se sobras de aproximadamente 5% do ofertado. As pesagens dos animais foram realizadas no início do período experimental e a cada 21 dias, sempre em jejum hídrico e alimentar de 14 horas. Foram mensurados o pH e os comprimentos das meias-carcaças no momento da saída das câmaras, após o processo de resfriamento. Da meia-carcaça esquerda foram medidas AOL, EGSO à altura da 12a costela. No registro do tempo gasto em alimentação, ruminação, ócio e ingestão de água foram adotadas observações visuais dos animais a cada cinco minutos, por quatro períodos integrais de 24 horas. Para determinação do pH das fezes e do amido fecal, amostras de fezes foram coletadas do reto de cada bovino. Houve diferença significativa (P<0,05) para as características de ganho de peso final e peso metabólico, sendo que os animais do tratamento DT+BIN peso final (498,43 kg e peso metabólico de 93,27 kg) e os animais do tratamento MGI peso final (481,67 kg e peso metabólico de 90,32 kg) foram mais eficientes do que os animais do tratamento DT com peso final (456,43 kg e peso metabólico de 88,52 kg). No desempenho de características de consumo, os animais que ingeriram as dietas DT+BIN (9,24 kg/dia) e MGI (7,34 kg/dia) foram mais eficientes do que os que consumiram a dieta DT (6,92 kg/dia). O consumo em percentagem do peso vivo do tratamento DT+BIN foi maior em 20,99% em analogia ao tratamento MGI e 26,59% ao DT. Na avaliação da carcaça houve diferença significativa (P<0,05) somente para os atributos, peso de carcaça quente (PCQ) e espessura de gordura subcutânea subjetiva (EGSS). O PCQ do tratamento DT+BIN (281,81 kg) foi superior ao tratamento DT (238,20 kg), e não diferiu do tratamento MGI (268,98 kg). A característica EGSS foi superior para o tratamento DT+BIN em 12,36% quando confrontado ao tratamento MGI e 50% mais elevado quando cotejado ao tratamento DT. Houve diferenças nos tempos usados em mastigação, ruminação, ócio e ingestão de água entre os tratamentos (P<0,05), exceto para o tempo gasto em alimentação. Os tempos empregados em ruminação e mastigação foram maiores para o tratamento DT+BIN (181,20 e 278,40 min/dia), porém, quanto aos tratamentos MGI (94,20 e 171 min/dia) e DT (60 e 148,20 min/dia) não ocorreram diferenças (P>0,05). Os tempos de ócios e ingestões de água diferiram entre si, com valores de 436,80 e 4,46 min/dia para o tratamento DT+BIN, 546,0 e 2,81 min/dia para o tratamento MGI e 568,80 e 4,02 min/dia para o tratamento DT. Os valores médios em percentagem do amido fecal, matéria seca fecal, medida de pH para o local de fermentação do amido e consumo de amido/kg não foram influenciados (P>0,05) pelos tratamentos. O escore de consistência fecal e FDN das fezes foram influênciados (P<0,05) pelos tratamentos com valores de escore e FDN fecal de 3,12 e 46,29% para o tratamento DT+BIN, 2,92 e 35,65% para o tratamento MGI e 3,20 e 48,97% para o tratamento DT. A adição de 10% de BIN em dieta totalmente concentrada, promove melhoria no consumo de MS, g/kg de PV, % de MS em relação ao PV, maior peso e acabamento de carcaça. A dieta de alto grão (MGI) foi superior a dieta de alta proporção de concentrado (DT) para a característica de ganho de peso final aos 84 dias e peso de carcaça quente. O tempo gasto com ruminação, mastigação e a eficiência de alimentação (g FDN/h) e (g de FDNfe/h) foi mais alto para a dieta com adição de BIN. A consistência de fezes avaliada nos bovinos do tratamento DT+BIN com 90% de concentrado e 10% de BIN classificada como firme evidencia que estes animais tiveram uma melhor fermentação ruminal

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A Esquistossomose à uma doenÃa causada por parasitos do gÃnero Schistosoma matando centenas de milhares de pessoas por ano no mundo. O Schistosoma tem vÃrias espÃcies com interesse clÃnico. No Brasil o causador da Esquistossomose à o S. mansoni e o principal hospedeiro e reservatÃrio do parasito à o homem sendo a partir desse que os ovos sÃo eliminados nas fezes. Os hospedeiros intermediÃrios sÃo os caramujos. As principais espÃcies de caramujos hospedeiros do Schistosoma mansoni no Brasil sÃo: Biomphalaria glabrata, Biomphalaria tenagophila e Biomphalaria straminea. Sendo a terceira espÃcie a predominante no CearÃ. A doenÃa tem presenÃa constante em mais de 74 paÃses (praticamente todos subdesenvolvidos). Cerca de 500 a 600 milhÃes de pessoas correm riscos de serem atingidas e mais de 200 milhÃes sÃo infectadas a cada ano, e isso se deve principalmente a falta de saneamento bÃsico e educaÃÃo sanitÃria. Para a melhor profilaxia desta doenÃa, deve ser feito o diagnÃstico e o tratamento da populaÃÃo, orientando para evitar entrar em contato com Ãguas que contenham caramujos (aÃudes, lagos, lagoas, rios etc). à uma doenÃa que pode evoluir para complicaÃÃes graves, eventualmente levando ao Ãbito em funÃÃo de extensa fibrose decorrente da presenÃa em parÃnquima hepÃtico de ovos do Schistosoma mansoni, formando granulomas. O principal objetivo desse estudo foi desenvolver uma estratÃgia para aumentar a eficÃcia na identificaÃÃo de pacientes infectados com S.mansoni, em Ãrea de baixa endemicidade, no Estado do CearÃ, usando um protocolo combinando uma tÃcnica sorolÃgica (IgG â ELISA) com exames de fezes seqÃenciais. Esse trabalho foi realizado em etapas, no qual na primeira, dos 287 indivÃduos que realizaram o mÃtodo de Kato-Katz, 11 (3,8%) apresentaram resultados positivos para S. mansoni. Com relaÃÃo aos outros helmintos foram encontrados: Trichuris trichiura 25 (8,7%), Ascaris lumbricoides 19 (6,6%) e Ancilostomideos 15(5,2%). Em relaÃÃo ao testes de ELISA, 97 (33,8%) foram positivos. Todos os pacientes que apresentaram ovos nas fezes foram positivos no teste sorolÃgico. Neste grupo estÃo inclusos os 11 que foram positivos na coproscÃpia. Dos pacientes com ELISA positivo e Kato-Katz negativos, apenas 56 entregaram as trÃs amostras de fezes para uma segunda anÃlise coproscÃpica. Desses, 14 (25%) foram positivos para Schistosoma mansoni. Dos 42 pacientes que continuavam negativos, 22 responderam no questionÃrio que nunca tiveram esquistossomose como tambÃm nunca receberam tratamento para a doenÃa. O presente estudo nÃo trata sà de determinar a prevalÃncia da doenÃa no municÃpio, e sim de identificar o maior nÃmero possÃvel de indivÃduos infectados, usando o mÃtodo sorolÃgico que foi aplicado de forma a contemplar a populaÃÃo residente em Ãreas de risco de transmissÃo ou expostas ao risco de infecÃÃo, principalmente por atividades domÃsticas e de lazer. Diante destes resultados, acredita-se, em concordÃncia com outros autores, que utilizando a tÃcnica de ELISA combinado com anÃlises repetidas de no mÃnimo 5 lÃminas de fezes, torna-se mais fÃcil diagnosticar pacientes com a esquistossomose, melhorando assim a hipÃtese que provavelmente em um futuro prÃximo, seremos capazes de combinar mÃtodos parasitolÃgicos e sorolÃgicos no programa de controle da esquistossomose, um fator importante para detecÃÃo de novos portadores da doenÃa. / Schistosomiasis is a disease caused by parasites of the genus Schistosoma, killing hundreds of thousands of people each year worldwide.. The Schistosoma has several species of clinical interest. In our country the cause of Schistosomiasis is the S. mansoni and the main reservoir host and the parasite is starting with the man that the eggs are removed in feces. The snails are the intermediate host. The main species of snails host of Schistosoma mansoni in Brazil are: Biomphalaria glabrata, Biomphalaria tenagophila and Biomphalaria straminea. The third kind in the predominant in CearÃ. The disease has presence in over 74 countries (virtually all underdeveloped). Around state 500 to 600 million people are at risk of being affected and more than 200 million are infected every year, and this is mainly due to lack of sanitation and health education. To the best prevention of this disease is to be made the diagnosis and treatment of the population to avoid targeting comes in contact with water containing snails (ponds, lakes, rivers etc). It is a disease that can develop into serious complications, possibly leading to death according to extensive fibrosis caused by the presence in liver parenchyma of the Schistosoma mansoni eggs, forming granulomas. So the main objective of this study was to develop a strategy to increase effectiveness in identifying patients positive for Schistosomiasis in areas of low endemic in the state of Ceara, using a protocol combining a technique in which antibodies (IgG - ELISA) with examinations of sequential stool. This work was followed by steps, in which the first of the 287 patients who underwent the method of Kato-Katz, 11 (3.8%) showed positive results for S. mansoni. On the other helminths are: Trichuris trichiura 25 (8.7%), A. lumbricoides 19 (6.6%) and Hookworms 15 (5.2%). For the tests, ELISA, 97 (33.8%) were positive. All patients who had eggs in the feces were positive in the serologic test. In this group are included the 11 that were positive in feces analysis (Figure 1). From patients with Elisa positive and negative Kato-Katz, only 56 handed the three samples of stool for a second analysis Of these, 14 (25%) were positive for Schistosoma mansoni. Of the 42 patients who remained negative, 22 responded in the questionnaire that had never schistosomiasis but never received treatment for the disease. Our present study was to not only determine the prevalence of the disease in the municipality, and to identify the largest possible number of infected individuals, the serological method was applied in order to accommodate the population living in those areas of risk of transmission or at risk of infection, mainly by domestic and leisure activities. In view of our results, we believe, in agreement with other authors, that using the ELISA technique combined with repeated analysis of at least 5 simples of feces, it becomes easier to diagnose patients positive for schistosomiasis, thus improving the hypothesis that probably in the near future, being able to combine parasitological and sorological in the programme for the control of schistosomiasis, an important factor for detection of new carriers of the disease.

Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-05-11T14:59:51Z No. of bitstreams: 2 Tese_Andrelisse Arruda_2017.pdf: 5916961 bytes, checksum: d3d6f3dd277d94f02ba45cb40e90dc81 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-05-11T15:00:15Z (GMT) No. of bitstreams: 2 Tese_Andrelisse Arruda_2017.pdf: 5916961 bytes, checksum: d3d6f3dd277d94f02ba45cb40e90dc81 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-05-11T15:00:15Z (GMT). No. of bitstreams: 2 Tese_Andrelisse Arruda_2017.pdf: 5916961 bytes, checksum: d3d6f3dd277d94f02ba45cb40e90dc81 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-10-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Microrganisms living in insects’ midgut have been isolated and identified for developing biotechnological tools to fight vector-born diseases. In this context, the mosquitoes Anopheles from different regions around the world have been studied about their midgut microbiota focused on paratransgenesis. However, information about microrganisms living in neotropical mosquitoes midgut are scarce. And, specially about Anopheles darlingi, the main malaria vector in the Brazilian Amazon, still this study, there were not any report about culturable microrganisms associated to this insect. The first step for paratransgenesis is to isolate culturable microrganisms naturally associated to the insect vector, and thus amenable to experimentation in laboratory. The objectives of this work were to isolate and to identify culturable bacteria and yeasts isolated from feces of Anopheles darlingi, the main vector of malaria in Brazil; to estimate the species richness and frequency distribution of the sampled bacteria and yeasts and to characterize and to select the sampled bacteria and yeasts isolated from feces of An. darlingi with potential for paratransgenesis. The female mosquitoes of An.darlingi were captured in two rural places of Porto Velho, Rondônia, Brasil. For improving the bacterial growth, mosquito feces were collected on LB agar medium and cultivated at 37 ºC for 24 hours, and for improving the yeast growth, mosquito feces were collected on YPD agar medium with Chloramphenicol and cultivated at 30 ºC for 48 hours. Sixty pure bacteria colonies and sixty pure yeast colonies were sampled. The isolates were preserved in -80 ºC freezer. PCR reactions with genomic DNA from each isolate were perfomed using the primers of 16S rRNA genes for bacteria and 26S and ITS for yeasts. From 60 bacterial isolates, 55 samples were identified. From 60 yeast isolates, 27 samples were identified. The fragments were sequenced with the Sanger method and the sequences with similarities above of 97% with sequences in reference database were deposited in Genbank (NCBI). For bacteria, MALDI-TOF, VITEK®2 and BBL Crystal were also used as a complementar protocols to identify the isolates. The identified bacteria fall into 8 genera, Enterobacter, Klebsiella, Cedecea, Pantoea, Serratia, Acinetobacter, Burkholderia and Staphylococcus. The identified yeast fall into 7 genera, Pseudozyma, Papiliotrema (Cryptococcus), Meyerozyma (=Pichia), Rhodotorula, Candida, Hanseniaspora and Metschnikowia. As candidates to paratransgenesis to control of malaria in An. darlingi are those bacteria belonging to the genera Pantoea and Serratia and the yeasts belonging to the genera Meyerozyma (=Pichia), Pseudozyma, Hanseniaspora and Metschnikowia. / Microrganismos contidos no trato digestório de insetos vem sendo isolados e identificados com o intuito de desenvolver ferramentas biotecnológicas para o controle de doenças transmitidas por insetos. Nesse contexto, mosquitos Anopheles de diferentes partes do globo têm sua microbiota investigada com foco em paratransgênese. No entanto, a informação sobre microrganismos associados aos anofelinos neotropicais é escassa. Tratando-se de Anopheles darlingi, o principal vetor de malária no Brasil, até o presente trabalho, não havia informações sobre microrganismos cultiváveis associados a esse vetor. Os objetivos deste trabalho foram isolar e identificar bactérias e leveduras cultiváveis isoladas das fezes de An. darlingi, o principal vetor da malária no Brasil; estimar riqueza e distribuição de frequência das bactérias e leveduras amostradas, e caracterizar e selecionar bactérias e leveduras isoladas das fezes de An. darlingi com potencial para paratransgênese. Os mosquitos An. darlingi fêmeas foram coletados em duas localidades rurais de Porto Velho, Rondônia, Brasil. Para favorecer o crescimento de bactérias, fezes dos mosquitos foram coletadas em meio LB ágar e cultivadas à 37°C por 24 horas, e para propiciar o crescimento de leveduras, fezes dos mosquitos foram coletadas em meio YPD ágar com cloranfenicol e cultivadas à 30°C por 48 horas. Sessenta colônias bacterianas e 60 colônias leveduriformes foram amostradas. Os isolados foram preservados em freezer -80 °C. Foram realizadas PCR utilizado DNA genômico dos isolados com iniciadores para a região do DNA ribossomal 16S para bactérias e 26S e ITS para leveduras. Todas as 60 bactérias isoladas foram identificadas. Das 60 leveduras isoladas, 27 foram identificadas. Os fragmentos foram sequenciados pelo método Sanger e as sequências com similaridades superiores a 97% frente a sequências disponíveis em bancos de dados foram depositadas no GenBank. Para bactérias, MALDI-TOF, VITEK®2 e BBL Crystal foram utilizados como métodos complementares para identificação dos isolados. As bactérias identificadas pertencem a 8 gêneros: Staphylococcus, Burkholderia, Cedecea, Enterobacter, Klebsiella, Pantoea, Serratia e Acinetobacter. As leveduras identificadas pertencem a 7 gêneros: Candida, Meyerozyma (=Pichia), Metschnikowia, Hanseniaspora, Rhodotorula, Papiliotrema (=Cryptococcus) e Pseudozyma. São candidatas à paratransgênese para o controle da malária em An. darlingi as bactérias do gênero Pantoea e Serratia e as leveduras dos gêneros Meyerozyma (Pichia), Metschkowia, Hanseniaspora e Pseudozyma.

Este estudo teve como objetivo avaliar a utilização da enzima fitase e de minerais orgânicos (Cu e Zn) na dieta de suínos visando a redução do poder poluente dos dejetos. Foram utilizados 16 suínos com idade média de 68 dias, alocados na câmara climática, em gaiolas de metabolismo para coleta de fezes e urina. Os animais foram mantidos em duas faixas de temperaturas ambientais: conforto térmico e estresse térmico, e submetidos aos tratamentos: controle (T1); suplementação com minerais orgânicos (T2); suplementação com fitase (T3); suplementação com minerais orgânicos e fitase (T4). As fezes e a urina foram analisadas quanto à concentração de P, N, Na, K, Cu, Zn, Ca. Mensurou-se também temperatura retal e superficial dos animais, volume de fezes e urina excretadas, assim como o consumo de água e ração. Observou-se que o estresse térmico afetou a temperatura retal e superficial, além do volume de fezes excretadas (P<=0,05). A excreção de Zn e Ca foram reduzidas com a utilização da enzima fitase, enquanto que o Cu e Zn orgânicos beneficiaram o Zn, Ca e P (P<=0,05). O estresse térmico aumentou significativamente a excreção de Cu, enquanto a de Na foi reduzida (P<=0,05). Embora não tenha ocorrido interação na atuação da enzima fitase e dos minerais orgânicos, estes aditivos contribuíram com a redução da excreção de minerais. / This study aimed to evaluate the use of phytase and organic minerals (Cu and Zn) in pig diets in order to reduce the power of polluting waste. Sixteen pigs at the age of 68 days, were allocated in metabolism studies cages for collection of feces and urine, in a climatic chamber. The animals were kept in two tracks of ambient temperatures: thermal comfort and heat stress, and subjected to the treatments: control (T1); organic minerals supplementation (T2); phytase supplementation (T3); organic minerals and phytase supplementation (T4). Feces and urine were analyzed for P, N, Na, K, Cu, Zn and Ca concentrations. It was also measured rectal temperature, body surface temperature, feces and urine volume and the food and water consumption. The results had shown that heat stress affected the rectal and superficial temperature, and excreted feces volume (P<=0,05). The Zn and Ca excretion were reduced with the phytase use, whereas organic Cu and Zn benefited Zn, Ca and P (P<=0,05). The heat stress significantly increased Cu excretion, while Na was reduced (P<=0,05). Although there was no interaction on the activity of phytase and organic minerals, these additives contributed to the excretion reduction of minerals.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-06-22T18:03:31Z No. of bitstreams: 1 2016 - Caroline Spitz dos Santos.pdf: 4739307 bytes, checksum: 0073345f08ba619e059be0cc4a39b481 (MD5) / Made available in DSpace on 2017-06-22T18:03:31Z (GMT). No. of bitstreams: 1 2016 - Caroline Spitz dos Santos.pdf: 4739307 bytes, checksum: 0073345f08ba619e059be0cc4a39b481 (MD5) Previous issue date: 2016-02-26 / CAPES / The study on parabasalids in companion animals is gaining more attention for its association with diarrhea. The flagellate Pentatrichomonas hominis has been reported in association with domestic cats since the early 20th century. As a eurixenic parasite has been described in several hosts, including humans, dogs, bovines, rats, and a variety of wild animals. Another parabasalid with great importance in livestock, Tritrichomonas foetus has been also described in cats, pigs, and humans. These observations raise questions about the zoonotic potential of both species and their host specificity. This study aimed to diagnose parabasalid species found in cats using morphological and molecular analysis. Therefore, this study was divided into two parts to assess two different cat populations. In the first part, 41 animals in a trial cattery were evaluated. Twenty-six percent of the animals (11) were positive for P. hominis, at both techniques as fresh examination and culture. The DNA was extracted from the samples in culture and rRNA genes were amplified by polymerase chain reaction (PCR) using universal primers (TFR1 and 2) and other two specific species for T. foetus (TFR3 and 4) and P. hominis (TH3 and 5). Morphological analysis of trophozoites revealed the presence of five previous plagues core round and axostyle tapering uniform, characteristic way of P. hominis. The results of morphology were confirmed by molecular study. The sequencing of the isolates revealed a sequence with 100% similarity to P. hominis isolated from cats and dogs deposited in Genbank. This is the first study in Brazil with pointed out the presence of Parabasalids in cats by using morphological and molecular data and it is the one in the literature where P. hominis was isolated. In the second part of this study, 77 samples of feces from cats from the clinical care of HVPA-UFRRJ were examined. Only four of 77 samples tested were positive. Morphological analysis showed predominantly pear-shaped protozoa with three previous scourges, elongated nucleus and ax?stilo abruptly ending in characteristic fillet in T. fetus. In scanning electron microscopy and transmission, were visualized the identifying characters were similar to those previously reported for T. foetus. Molecular analysis confirmed the morphological diagnosis in the organism from four samples showed a sequence with 99.7 to 100% of similarity. It was deposited in Genbank as T. foetus. Despite the morphological analysis have recognized only T. foetus in the four samples examined, three of them were also positive for P. hominis in molecular analysis used as a differential diagnosis using species-specific primers (TH3 and TH5). The molecular analysis was used as a confirmatory tool for the presence of only one species present in evaluated feces. This demonstrated that not only T. foetus was identified in this study, but a co-infection by P. hominis cats could be considered. This indication was only confirmed as a diagnostic techniques when the morphological analysis and molecular biology were used to confirm both species. / O estudo sobre parabasal?deos em animais de companhia vem ganhando cada vez mais aten??o por sua associa??o a quadros de diarreia. O flagelado Pentatrichomonas hominis foi relatado em associa??o com gatos dom?sticos desde o in?cio do s?culo 20. Por ser um parasito eurix?nico, j? foi descrito em diversos hospedeiros, incluindo os seres humanos, c?es, bovinos, ratos e uma variedade de animais selvagens. Assim tamb?m outra esp?cie de parabasal?deo com grande import?ncia na pecu?ria, Tritrichomonas foetus j? foi descrito em gatos, su?nos, e em humanos tamb?m. Tais observa??es levantam d?vidas sobre o potencial zoon?tico de ambas as esp?cies e sua inespecificidade hospedeira. Este estudo teve por objetivos diagnosticar esp?cies de parabasal?deos encontrados em gatos utilizando de an?lise morfol?gica e molecular. Para tanto, este estudo foi dividido em duas partes para avaliar duas popula??es felinas distintas. Na primeira parte, 41 animais de um gatil de experimenta??o foram avaliados. Vinte e seis por cento dos animais (11) foram positivos para P. hominis, tanto no exame a fresco quanto na cultura. O DNA foi extra?do das amostras em cultura e os genes de rRNA foram amplificados por rea??o em cadeia da polimerase (PCR), utilizando iniciadores universais (TFR1 e 2) e outros dois esp?cies espec?ficos para T. foetus (TFR3 e 4) e P. hominis (TH3 e 5). A an?lise morfol?gica dos trofozo?tos revelou a presen?a de cinco flagelos anteriores, n?cleo redondo e ax?stilo afunilando de maneira uniforme, caracter?stico de P. hominis. Os resultados da morfologia foram confirmados pelo estudo molecular. O sequenciamento dos isolados revelou 100% de similaridade de sequ?ncia com P. hominis isolado de gato e de c?o depositados no Genbank. Este ? o primeiro estudo realizado no Brasil sobre a presen?a de parabasal?deos em gatos utilizando dados morfol?gicos e moleculares e o ?nico na literatura onde somente P. hominis foi isolado. Na segunda parte deste estudo, 77 amostras de fezes de gatos provenientes do atendimento cl?nico do HVPA-UFRRJ foram examinadas. Somente quatro amostras apresentaram positivas. A an?lise morfol?gica demonstrou protozo?rios predominantemente piriformes com tres flagelos anteriores, n?cleo alongado e ax?stilo terminando bruscamente em filete caracter?stico de T. foetus. Na microscopia eletr?nica de varredura e de transmiss?o, foram visualizados caracteres de identifica??o semelhantes aos descritos na literatura para T. foetus. ? an?lise molecular, confirmou o diagn?stico morfol?gico nas quatro amostras, e no sequenciamento apresentaram 99,7-100% de similaridade com sequencias de T. foetus depositadas no Genbank. Apesar da an?lise morfol?gica ter reconhecido somente T. foetus nas quatro amostras, tr?s delas foram positivas tamb?m para P. hominis na an?lise molecular utilizada como diagn?stico diferencial utilizando iniciadores esp?cie-espec?fica (TH3 e TH5). O estudo molecular foi utilizado como ferramenta confirmat?ria da presen?a de somente uma esp?cie presente nas fezes avaliadas. Isso demonstra que n?o s? T. foetus foi identificado pelo presente estudo, como tamb?m a coinfec??o por P. hominis em felinos. Estas informa??es s? foram confirmadas quando se utilizou as t?cnicas de diagn?stico em conjunto como an?lise morfol?gica simples e biologia molecular.

The ultimate objective of this work was to develop a real-time method for detecting and monitoring β-D-galactosidase as a suitable indicator of the potential presence of total coliform bacteria in water environments. Preliminary comparison of the chromogenic substrate, chlorophenol red β-D-galactopyranoside and the fluorogenic substrate, MuGAL, revealed unreliable results with the fluorogenic technique due to interference from compounds commonly found in environmental water samples. Thus, the chromogenic assay was further explored. Hydrolysis of the chromogenic substrate chlorophenol red β-D-galactopyranoside by β-D-galactosidase to yield chlorophenol red was the basis of this assay. Fundamental studies with chlorophenol red β-Dgalactopyranoside showed that β-D-galactosidase occurs extracellularly and in low concentrations in the polluted water environment. A direct correlation between enzyme activity and an increase in environmental water sample volume, as well as enzyme activity with total coliform colony forming unit counts were observed. Spectrophotometric detection was achieved within a maximum period of 24 h with a limit of detection level of 1 colony forming unit 100 ml[superscript -1]. This enzyme also exhibited physical and kinetic properties different from those of the pure commercially available β-D-galactosidase. Cell permeabilisation was not required for releasing enzymes into the extracellular environment. PEG 20 000 offered the best option for concentrating β-D-galactosidase. The source of β-D-galactosidase in the polluted environmental water samples was confirmed as Escherichia coli through SDS-PAGE, tryptic mapping and MALDI-TOF, thus justifying the further use of this method for detecting and/or monitoring total coliforms. Several compounds and metal ions commonly found in environmental water samples (as well as those used in water treatment processes) did have an effect on β-D-galactosidase. All the divalent cations except Mg [superscript 2+], at the concentrations studied, inhibited the relative activity of β-D-galactosidase in both commercial β-D-galactosidase and environmental samples. Immobilisation of chlorophenol red β-D-galactopyranoside onto a solid support material for the development of a strip bioprobe was unsuccessful, even though the nylon support material yielded some positive results. A monthly (seasonal) variation in β-Dgalactosidase activity from the environmental water samples was observed, with the highest activity coinciding with the highest monthly temperatures. Electro-oxidative detection and/or monitoring of chlorophenol red was possible. Chlorophenol red detection was linear over a wide range of concentrations (0.001-0.01 μg ml[superscript -1]). Interference by chlorophenol red β-D-galactopyranoside in the reduction window affected analysis. A range of phthalocyanine metal complexes were studied in an attempt to reduce fouling and/or increase the sensitivity of the biosensor. The selected phthalocyanine metal complexes were generally sensitive to changes in pH with a reduction in sensitivity from acidic pH to alkaline pH. The tetrasulphonated phthalocyanine metal complex of copper was, however, more stable with a minimum change of sensitivity. The phthalocyanine metal complexes were generally stable to changes in temperature. While only two consecutive scans were possible with the unmodified glassy carbon electrode, 77 consecutive scans were performed successfully with the CuPc-modified glassy carbon electrode. Among the phthalocyanine metal complexes studied, the CuPc-modified glassy carbon electrode therefore provided excellent results for the development of a biosensor. The CuPc modified-glassy carbon electrode detected 1 colony forming unit 100 ml[superscript -1] in 15 minutes, while the plain unmodified glassy carbon electrode required 6 hours to detect the equivalent number of colony forming units. CoPc, ZnPc and CuTSPc required 2, 2.25 and 1.75 h, respectively, to detect the same numbers of colony forming units. The CuPcmodified glassy carbon electrode detected 40 colony forming units 100 ml[superscript -1] instantly. In general, a direct correlation between colony forming units and current generated in the sensor was observed (R2=0.92). A higher correlation coefficient of 0.99 for 0-30 coliform colony forming units 100 ml[superscript -1] was determined. Current was detected in some water samples which did not show any colony forming units on the media, probably due to the phenomenon of viable but non-culturable bacteria, which is the major disadvantage encountered in the use of media for detecting indicator microorganisms. This novel biosensor therefore presents a very robust and sensitive technique for the detection and/or monitoring of coliform bacterial activity in water.

Microbial contamination of drinking water poses an important health risk which causes severe illnesses and epidemics. In order to improve surface and drinking water quality, the understanding of fecal pathogen contamination processes including their prevention and control needs to be enhanced. The watershed model soil water assessment tool (SWAT) is commonly used to simulate the complex hydrological, meteorological, erosion, land management and pollution processes within river basins. In recent years, it has been increasingly applied to simulate microbial contamination transport at the watershed scale. SWAT is used in this study to simulate Escherichia coli (E.coli) and fecal coliform densities for the agriculturally dominated Payne River Basin in Ontario, Canada. Unprecedented extensive monitoring data that consist of 30 years of daily hydrological data and 5 years of bi-weekly nutrient data have been used to calibrate and validate the presented model here. The calibration and validation of the streamflow and nutrients indicate that the model represent these processes well. The model performs well for periods of lower E. coli and fecal coliform loadings. On the other hand, frequency and magnitude of higher microbial loads are not always accurately represented by the model.