Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-10-04T13:00:01Z No. of bitstreams: 1 2016 - Thais Paes Ferreira.pdf: 1131707 bytes, checksum: c98267296d6293f703188fe1291622db (MD5) / Made available in DSpace on 2016-10-04T13:00:01Z (GMT). No. of bitstreams: 1 2016 - Thais Paes Ferreira.pdf: 1131707 bytes, checksum: c98267296d6293f703188fe1291622db (MD5) Previous issue date: 2016-05-11 / Fluazuron is a benzoylphenylurea acarine growth inhibitor that is marketed for the control of cattle tick. Although the growth regulators are widely studied class, the number of papers describing analytical methods for determining the fluazuron in plasma samples is reduced. The development of a simple and fast analytical method for quantifying fluazuron in bovine plasma by HPLC-UV enables the evaluation of drug plasma profile and can be applied on bioavailability analysis of formulations containing fluazuron. In recent years there has been a technological breakthrough for chromatographic instrumentation, providing rapid, robust and sensitive methods, but the pretreatment of the sample stage becomes the limiting factor in this process. New extraction methods have been developed, however are often complex, expensive methods and are not as sophisticated to handle complex matrices, such as plasma. The aim of this study was a comparative study of three extraction methods: LLE, SPE and matrix solid phase dispersion (MSPD). Pooled plasma samples were used as biological material, partly as raw samples and partly spiked with concentrations of fluazuron. For each extraction method (with exception of DMFS) was conducted using various solvents (ethyl acetate, dichloromethane, diethyl ether, hexane and acetonitrile) in order to evaluate the best extractor solvent. After the extractions the samples were concentrated and analyzed by HPLC/UV. The chromatographic separation was achieved on Kromasil C18 column preceded by guard column of matching chemistry, with mobile phase of acetonitrile: water (80:20, v/v) at flow rate of 1.0 mL/min. The comparative analysis of the extraction procedures was based on selectivity, precision and accuracy of the method. Results showed that LLE not presented accuracy (< 80 and >120%) and precision (CV > 15%) with any solvent tested. SPE method showed good accuracy (80-120%) and precision (CV < 15%) for ethyl acetate, dichloromethane and diethyl ether. MSPD method using ethyl acetate solvent showed good accuracy and precision. However, SPE also allows a lower solvent consumption and shorter analysis time. The validation of the analytical method showed linearity, selectivity, precision, accuracy and, absence matrix effects and residual, thus proving it as suitable for routine analysis; This method showed to be an important investigative tool in the analysis of fluazuron plasma concentration in cattle. Fluazuron topical administration in bovine reached the systemic circulation (Cmax=62,8 ng/mL), was absorbed (tmax=48 hs), while maintaining quantifiable blood plasma levels for up to 14 days after the treatment with a 2,5 mg/Kg dosage. / Fluazuron ? um inibidor do crescimento da classebenzoilfenilur?ia, sendo comercializado para o controle de carrapatos do gado. Embora a classe de reguladores de crescimento sejam amplamente estudada, o n?mero de artigos que descrevem os m?todos anal?ticos para a determina??o do fluazurom em amostras de plasma ? escasso. O desenvolvimento de um m?todo anal?tico simples e r?pido para quantificar fluazuron em plasma bovino por CLAEUV permite a avalia??o do perfil plasm?tico do f?rmaco e pode ser aplicado na an?lise de biodisponibilidade de formula??es contendo fluazurom. Nos ?ltimos anos, tem acontecido um avan?o tecnol?gico para a instrumenta??o cromatogr?fica, proporcionando m?todos r?pidos, eficazes e sens?veis, mas a etapa de pr?-tratamento da amostra torna-se o fator limitante neste processo. Novos m?todos de extra??o t?m sido desenvolvidos, no entanto, muitas das vezes s?om?todos complexos, caros e n?o t?o sofisticados para matrizes complexas como o plasma. O objetivo deste estudo foi um estudo comparativo de tr?s m?todos de extra??o: extra??o l?quido-l?quido (ELL), extra??o em fase s?lida (EFS) e dispers?o de matriz em fase s?lida (DMFS). Ums mistura de amostras de plasma foi utilizada como material biol?gico, uma parte como amostra bruta e outra parcialmente enriquecida com concentra??es de fluazuron. Para cada m?todo de extra??o (exceto DMFS) foram analisadosdiversos solventes (acetato de etila, diclorometano, ?ter diet?lico, hexano e acetonitrila), a fim de avaliar o melhor solvente extrator. Ap?s as extra??es, as amostras foram concentradas e analisadas por CLAE/UV. A separa??o cromatogr?f ica foi obtida na coluna Kromasil C18 precedida de pr?-coluna de fase qu?mica correspondente, com fase m?vel de acetonitrila: ?gua (80:20, v/v) a um fluxo de 1,0 mL /min. A an?lise comparativa dos procedimentos de extra??o foi baseada na seletividade, precis?o e exatid?o do m?todo. Os resultados mostraram que a precis?o ELL n?o apresentou exatid?o (<80 e> 120%) e precis?o (CV> 15%) com nenhum dos solventes analisados. O m?todo EFS mostrou boa exatid?o (80-120%) e precis?o (CV <15%) para o acetato de etila, diclorometano e ?ter diet?lico. O m?todo de DMFS utilizando o solvente acetato de etila mostrou boa exatid?o e precis?o. No entanto, a EFS tamb?m permite um menor consumo de solventes e um tempo curto de an?lise; A valida??o do m?todo anal?tico mostrou linearidade, seletividade, precis?o, exatid?o e, aus?ncia de efeitos de matriz e residual, demonstrando-se adequado para an?lises de rotina; Este m?todo mostrou-se uma ferramenta de investiga??o importante na an?lise de concentra?? plasm?tica em bovinos. Fluazuron administrado por via t?pica em bovinos atingiu a circula??o sist?mica (Cmax = 62,8 ng /ml) e foi absorvido (Tm?x = 48 Hs), se mantendo quantificav?l em n?veis plasm?ticos por at? 14 dias ap?s o tratamento com uma dosagem de 2,5 mg / kg.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-04-20T11:39:08Z No. of bitstreams: 1 2012 - Vanessa Paulino da Cruz Vieira.pdf: 1181202 bytes, checksum: 7bfcf1b5c3509ddacbb033dcaf5448c3 (MD5) / Made available in DSpace on 2017-04-20T11:39:08Z (GMT). No. of bitstreams: 1 2012 - Vanessa Paulino da Cruz Vieira.pdf: 1181202 bytes, checksum: 7bfcf1b5c3509ddacbb033dcaf5448c3 (MD5) Previous issue date: 2012-03-29 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / This study was conducted to evaluate the activity of fluazuron administered orally in the control of R. sanguineus in dogs. A total of 12 beagle dogs which, on day 0, treatment day, were divided into two groups: Group 1. Six animals treated with oral formulation fluazuron at a dose rate of 20 mg / kg body weight and Group 2. Six dogs kept as control without treatment. On the same day, containment devices for ticks denim cloth were attached with glue to the back trichotomized dogs. After this procedure, three challenges were performed on days +1, +20 and +40, where the animals of both groups received infestations with the three stage of the tick R. sanguineus: approximately 2.500 larvae, 200 nymphs and 25 adult couples. Were used a climatic chamber with biochemical oxygen demand, BOD, at a temperature of 27 ? 1 ? C and relative humidity of 80 ? 10%. From the fourth to tenth day after each challenge, the devices were opened daily to the collection of engorged stages. Larvae and nymphs were counted and divided into syringes for evaluating the changes. The engorged females were collected and fixed in petri dishes to perform the posture that was weighed and placed in syringes, sealed with cotton, to calculate the hatchability. The efficacy of recovery fluazuron on larvae of A. sanguineus in dogs was 84.3% for day of challenge of the day +20 and 36% for the day +40. With regard to the recovery of engorged nymphs of R. sanguineus in dogs fluazuron achieved an efficacy of 82.4% and 51.7% on days +20 and +40. There was statistical difference between the mean number of engorged nymphs of R. sanguineus recovered on days +20 and +40 (p ? 0.05). The efficacy of recovery of the fluazuron engorged females was lower than 30.3% over the whole experimental period. The efficacy of treatment on reproductive performance of R. sanguineus was less than 12.4% in the three days of challenge. In inhibiting the change of larvae or nymphs ecdysis, fluazuron showed the effectiveness of 0%, 96.9% and 45.0% for days +1, +20 and +40, respectively. On day +20, there was a statistically significant difference between the mean engorged larvae to nymphs who underwent changes (p ? 0.05). The effectiveness of fluazuron on the inhibition of molting or ecdysis of nymphs to adults was 0%, 99.5% and 63.5% for days +1, +20 and +40, respectively, a statistically significant difference between mean the control and treated groups on days +20 and +40 (p ? 0.05). The growth regulator arthropod fluazuron is effective in helping control larvae and nymphs of R. sanguineus, when administered orally to dogs at a dose of 20mg/Kg. The same dose and route of administration, has no significant negative effect on female reproductive R. sanguineus. / O presente trabalho foi realizado com o objetivo avaliar a atividade do fluazuron administrado por via oral no controle de R. sanguineus em c?es. Foram utilizados 12 c?es da ra?a Beagle, que, no dia 0, dia do tratamento, foram divididos em dois grupos: Grupo 1. Seis c?es tratados com formula??o oral de fluazuron na dosagem de 20 mg/Kg de peso corporal e Grupo 2. Seis c?es mantidos como controle, sem tratamento. No mesmo dia, dispositivos para conten??o de ixod?deos de pano brim foram aderidos com cola ao dorso tricotomizado dos c?es. Ap?s esse procedimento, foram realizados tr?s desafios, nos dias +1, +20 e +40, onde os animais dos dois grupos receberam infesta??es com as tr?s fases do carrapato R. sanguineus: aproximadamente 2.500 larvas, 200 ninfas e 25 casais de adultos. Foi utilizada c?mara climatizada com demanda bioqu?mica de oxig?nio, tipo BOD, a uma temperatura de 27 ? 1oC e umidade relativa de 80 ? 10%. Do quarto ao d?cimo dia ap?s cada desafio, os dispositivos foram abertos diariamente para a coleta das fases ingurgitadas. As larvas e as ninfas foram contadas e separadas em seringas para a avalia??o da muda. As f?meas ingurgitadas foram coletadas e fixadas em placas de petri para realiza??o da postura que foi pesada e colocada em seringas, vedadas com algod?o, para o c?lculo da eclodibilidade. A efic?cia do fluazuron sobre a recupera??o de larvas de R. sanguineus em c?es foi de 84,3% para o desafio do dia +20 e 36% para o dia +40. Com rela??o ? recupera??o de ninfas ingurgitadas de R. sanguineus em c?es, o fluazuron obteve uma efic?cia de 82,4% e 51,7% nos dias +20 e +40. Houve diferen?a estat?stica entre os n?meros m?dios de ninfas ingurgitadas de R. sanguineus recuperadas nos dias +20 e +40 (p?0,05). A efic?cia do fluazuron sobre a recupera??o de f?meas ingurgitadas foi inferior a 30,3% durante todo o per?odo experimental. A efic?cia do tratamento sobre a efici?ncia reprodutiva de R. sanguineus se apresentou inferior a 12,4% nos tr?s dias de desafio. Na inibi??o da muda ou ecdise de larvas para ninfas, o fluazuron apresentou uma efic?cia de 0%, 96,9% e 45,0% para os dias +1, +20 e +40, respectivamente. No dia +20, observou-se diferen?a estat?stica significativa entre as m?dias de larvas ingurgitadas que realizaram muda para ninfas (p?0,05). A efic?cia do fluazuron sobre a inibi??o da muda ou ecdise de ninfas para adultos foi de 0%, 99,5% e 63,5% para os dias +1, +20 e +40, respectivamente, havendo diferen?a estat?stica significativa entre as m?dias dos grupos controle e tratado nos dias +20 e +40 (p?0,05). O regulador de crescimento de artr?podes fluazuron ? eficaz no aux?lio do controle de larvas e ninfas de R. sanguineus, quando administrado oralmente em c?es, na dose de 20mg/Kg. Na mesma dose e via de administra??o, n?o apresenta efeito negativo significativo na reprodu??o de f?meas de R. sanguineus.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-06-21T16:48:23Z No. of bitstreams: 1 2016 - Fabr?cio Nascimento Gaud?ncio.pdf: 2705400 bytes, checksum: 84083c56a169de3a2c732060527db477 (MD5) / Made available in DSpace on 2017-06-21T16:48:23Z (GMT). No. of bitstreams: 1 2016 - Fabr?cio Nascimento Gaud?ncio.pdf: 2705400 bytes, checksum: 84083c56a169de3a2c732060527db477 (MD5) Previous issue date: 2016-02-22 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The objective of this research was to evaluate the effect of the arthropod growth regulator fluazuron in the energetic metabolism of Rhipicephalus microplus. 6 stabled cattle divided into control and treated groups containing three animals each. A commercial formulation containing the active compound fluazuron was administered topically in the treated group. The engorged female ticks were collected from some days before (day 0) and after treatment of animals (4, 8, and 15 days). After collecting the engorged females, the following biological parameters were analyzed: weight of the engorged female, weight of the female after oviposition, laying period, laying weight, egg production rate, reproductive efficiency index, percentage of hatching and nutritional index. It was also evaluated the metabolic profile: in the hemolymph (concentration of glucose, total protein and organic acids - oxalic, pyruvic and lactic, enzymatic activity of LDH, ALT and AST); in the fat body (glycogen concentration and percentage of variation of neutral lipids); in eggs (concentration of glucose and total protein content, percentage change of neutral lipids and enzymatic activities of ALT and AST); and in the cuticle of engorged female ticks (percentage change of neutral lipids). Secondly, female immersion test was conducted to assess the activities of carboxylesterases enzymes, monooxygenases (MFO) and glutathione-S-transferase (GST) in sensitive strain of eggs and larvae (Porto Alegre, RS) compared with resistant strain (Jaguar, RS). It also assessed the detoxification enzymes in the fat body of engorged females of sensitive strain. The results showed that all the biological parameters showed lower results when compared to the control group after the exposure to fluazuron, except the weight of the female after oviposition. Although an increase was observed in the concentration of glucose in hemolymph on the 8th day and glycogen in the fat body in 15th day, there was no significant difference in the comparison of means between control and treated day-to-day. The concentration of lactic acid gradually increased in the treated group over time due to treatment, indicating increased anaerobic metabolism. There was no significant difference in the percentage change of neutral lipids in fat body, cuticle and eggs in any of the time intervals. No significant difference was observed in the activity of transaminases in the hemolymph, but it was observed in eggs after the treatment on the 8th day. The activity of MFO was higher in the comparison between the resistant and sensitive strains in eggs, but in larvae, the increase was observed in carboxylesterases in the resistant strain. In females, MFO showed high activity in the fat body as a function of fluazuron exposure, especially on day 4 in the treated group. Increases in transaminase activity and detoxification enzymes in females and eggs were associated with possible cell damage and trying to metabolism of active principle-tested. For the first time, a research about the metabolic effects of fluazuron has been conducted, focusing on different biochemical aspects, demonstrating that its effectiveness is related to a number of metabolic changes, leading to a parasite homeostasis breaking. / O objetivo desta pesquisa foi avaliar os perfis metab?lico-energ?tico de carrapatos Rhipicephalus microplus expostos ao fluazuron. 6 bovinos foram estabulados e divididos em grupos controle e tratado, contendo 3 animais cada. Uma formula??o comercial contendo o princ?pio-ativo fluazuron foi administrada por via t?pica nos animais do grupo tratado. F?meas ingurgitadas foram coletadas em dias determinados: antes (dia 0) e ap?s o tratamento dos animais (4, 8 e 15 dias). Ap?s a coleta das f?meas ingurgitadas, foram analisados os seguintes par?metros biol?gicos: peso da f?mea ingurgitada, peso da quen?gina, per?odo de postura, peso da postura, ?ndice de produ??o de ovos, efici?ncia reprodutiva, percentual de eclos?o e ?ndice nutricional. Foi avaliado tamb?m o perfil metab?lico: na hemolinfa (concentra??o de glicose, prote?nas totais e ?cidos org?nicos - ox?lico, pir?vico e l?tico, atividade enzim?tica de LDH, ALT e AST), no corpo gorduroso (concentra??o de glicog?nio e percentual de varia??o de lip?dios neutros), nos ovos (concentra??o de glicose, prote?nas totais, percentual varia??o de lip?dios neutros e atividade enzim?tica de ALT e AST), e na cut?cula das f?meas ingurgitadas (percentual de varia??o de lip?dios neutros). Num segundo momento, realizou-se teste de imers?o de f?meas ingurgitadas para posterior avalia??o das atividades das enzimas carboxilesterases, monooxigenases (MFO) e glutationa-S-transferase (GST) em ovos e larvas de diferentes cepas: sens?vel (Porto Alegre, RS) e cepa resistente (Jaguar, RS) ao fluazuron. Avaliou-se tamb?m as enzimas de detoxifica??o no corpo gorduroso de f?meas ingurgitadas de cepa sens?vel. Na an?lise dos resultados, todos os par?metros apresentaram resultados inferiores, quando comparados ao grupo controle, em fun??o da exposi??o ao fluazuron, exceto o peso da quen?gina. Embora tenha sido observado um aumento na concentra??o de glicose hemolinf?tica no 8? dia e glicog?nio no corpo gorduroso no 15?dia, n?o houve diferen?a significativa na compara??o das m?dias entre controle e tratado dia-a-dia. A concentra??o do ?cido l?tico apresentou aumento progressivo significativo no grupo tratado ao longo do tempo em fun??o do tratamento, indicando aumento do metabolismo anaer?bio. N?o houve diferen?a significativa para as varia??es percentuais de lip?dios neutros no corpo gorduroso, cut?cula e ovos em nenhum dos dias de avalia??o. Embora n?o tenha sido observada diferen?a significativa nas atividades das transaminases na hemolinfa, observou-se aumento significativo nos ovos ap?s o tratamento no 8?dia. A atividade da MFO foi maior, na compara??o entre as cepas resistente e sens?vel, nos ovos, e com rela??o ?s larvas; o aumento observado foi nas carboxilesterases da cepa resistente. Nas f?meas, a MFO apresentou atividade elevada no corpo gorduroso em fun??o da exposi??o ao fluazuron, principalmente no 4?dia no grupo tratado. Os aumentos na atividade das transaminases e das enzimas de detoxifica??o nas f?meas ingurgitadas e nos ovos foram associados a poss?veis danos celulares e ? tentativa de metaboliza??o do princ?pio-ativo testado. Pela primeira vez, foi realizado um estudo acerca dos efeitos metab?licos do fluazuron, enfocando diferentes aspectos bioqu?micos, evidenciando que a sua efic?cia est? relacionada a diversas altera??es metab?licas, levando a uma quebra da homeostase do carrapato