1000 resultados para enzimas digestivas
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O experimento foi conduzido com o objetivo de avaliar o efeito da adição de um compelxo multienzimático exógeno composto de amilase, protease, lipase e celulase, em rações de juvenis de tambaqui, sobre os coeficientes de digestibilidade aparente (CDa) da proteína bruta (PB), extrato etéreo (EE), carboidratos (ENN) e energia bruta (EB). O delineamento experimental foi inteiramente casualizado com quatro tratamentos (quatro níveis de inclusão de enzimas, 0,0; 0,05; 0,10; e 0,15 %), três repetições (no tempo) e 10 peixes por unidade experimental. Foram utilizados 40 juvenis de tambaqui, com peso médio de 155,0 ± 0,49 g, distribuídos em quatro tanques de alimentação de 500 l, recebendo refeições à vontade das 8 às 12h, a cada hora. Em seguida os animais foram transferidos para coletores de fezes (200 l), onde permaneceram até às 18h, sendo a coleta de dejetos realizada a cada hora. A determinação dos CDa foi realizada pelo método indireto, sendo utilizado como indicador externo 0,5% de óxido de cromo-III (Cr2O3) incorporado à ração. Os resultados demonstraram que a suplementação das dietas com enzimas exógenas para juvenis de tambaqui aumenta a digestibilidade aparente dos nutrientes e energia bruta, no nível de inclusão de 0,05% (P<0,05%).
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O objetivo deste trabalho foi avaliar o efeito da adição na ração das enzimas digestivas exógenas amilase, lipase e protease sobre o desempenho de tambaqui, em três experimentos independentes. Em cada experimento foi avaliado o efeito da inclusão de uma enzima na ração em diferentes porcentagens: 0,0, 0,05, 0,1 e 0,2%, cada uma com quatro repetições, com 28 dias de duração. Foram realizadas biometrias do peso inicial, para homogeneização da amostra, e do peso final a fim de verificar o efeito dos tratamentos sobre o desempenho dos peixes. Em cada unidade experimental foram estocados dez peixes, alimentados duas vezes ao dia (8 e 14h) até a saciedade aparente, com ração comercial extrusada com 28% de proteína bruta, triturada, adicionada de enzima e depois peletizada. Ao final do experimento, o ganho de peso, a conversão alimentar aparente e o crescimento específico dos peixes, apresentaram diferenças significativas entre os tratamentos. As enzimas exógenas amilase a 0,05% e lipase a 0,2% na ração influenciaram o desempenho zootécnico de juvenis de tambaqui. A adição de protease exógena não apresenta a mesma influência.
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Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests
Efeito do uso de probiótico sobre o desempenho e atividade de enzimas digestivas de frangos de corte
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Shrimp farming in Brazil is a consolidated activity, having brought economical and social gains to several states with the largest production concentrated in the northeast. This fact is also reflected in higher feed intake, necessitating a more efficient feed management. Currently, management techniques already foresee food loss due to molting. In this sense, studies relating shrimp s digestive physiology, molting physiology and behavioral response of shrimp feed can optimize the feed management. Thus, our study aimed to evaluate the behavioral response of the marine shrimp L. vannamei (Crustacea: Penaeidae) in accordance with the stages of moulting cycle and feeding schedules based on higher or lower activity of proteolytic digestive enzymes; also, to investigate the influence of feeding schedule on hepatosomatic index and non-specific and specific protease activity (trypsin). Experiments were carried out at the Laboratory of Shrimp Behavioral Studies at UFRN in partnership with the Laboratory of Enzimology UFPE. Juveniles of L. vannamei weighting 5.25 g (+ 0.25 g) were kept in aquaria at a density of 33 shrimp m -2. In the first experiment, shrimp were fed in the light phase or in the dark phase for 8 days; in the ninth day, the animals were observed for 15 minutes every hour during the 12 hours of each phase of the photoperiod. We recorded the frequency of inactivity, exploration, food intake, burrowing, swimming and crawling behavior. At the end of the 12th observation session, the shrimp were sacrified and classified by the method of setogenesis in the molt cycle stages A, B, C, D0, D1, D2 or D3. We found that the shrimp in A stage show high levels of inactivity. Moreover, the frequency of food intake was very low. The shrimp in D3 stage also had low food intake and high inactivity associated with elevated frequencies of burrowing. In the second experiment, shrimp were kept in physiological acclimation to experimental conditions for 28 days, distributed in 12 treatments in the light phase and 12 treatments in the dark phase. In the end, the animals were sacrified and dissected to assess non-specific and specific protease activity (trypsin) activity. In general, these parameters did not vary among animals fed in the light phase and those fed in the dark phase. However, significant differences were found in the activity of specific and nonspecific proteases in relation to food treatment. In the light phase, the major proteolytic activities converged to 10 hours after the start of the light phase, while the lowest activities converged to 6 hours after the beginning of this phase. In the dark phase, the highest enzyme activity converged to 12 hours after the onset of phase, while the lowest activities converged to 3 hours after the onset of phase. In the third experiment, we sought to evaluate the behavioral responses of shrimp in relation to dietary treatments based on higher or lower activity of proteolytic enzymes, considering the results of the second experiment. The behavioral categories observed were the same as the ones in the first experiment, with observations of 30 minutes (15min before and 15min after food supply). We found variation in behavioral responses as a function of the treatments, with greater intake of food in shrimp fed during the period of greatest activity of proteolytic enzymes, in the light phase. Thus we see that periodic events associated with the shrimp s physiology interfere in their behavioral responses, revealing situations that are more adjustable to the provision of food, and consequently optimizing feeding management
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It was evaluated the effect of the addition of glutamine, polyunsaturated fatty acids or cellular wall of yeast to the diet of weaned pigs on the activity of the pancreatic enzymes (lipase, amylase and trypsin) and the intestinal mucous membrane (dipeptidase, sucrase and maltase) and on the performance. Forty-five weaned pigs were used and distributed in a randomized block design, in factorial outline, with four diets (T1 - basal diet (BD); T2 - BR + 1% glutamine; T3 - BD + 0,2% cellular wall of yeast; T4 - BD + 5% fish oil) and two slaughter ages (seven and 14 days post weaning). The performance was measured in the first two weeks post-weaning. The addition of 1% glutamine in the diet of pigs increased the specific and total activity of the amylase, and total activity of the trypsin in the second week post weaning. The others supplements not change the activity of the digestive enzymes in the pigs. Also an increase was observed in the total activity of the lipase, and specific activity of the trypsin and maltase in function of the age post-weaning. In general, the activities of the digestive enzymes were correlated positively, except for the dipeptidase that was not correlated with any other enzyme. Positive correlation was observed between weight gain and activity of the lipase and of the amylase. The supplements included in the diet not influence the performance of weaned pigs.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Pós-graduação em Zootecnia - FMVZ
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Pós-graduação em Zootecnia - FCAV
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Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Pós-graduação em Zootecnia - FMVZ
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Dissertação para obtenção do Grau de Mestre em Tecnologia e Segurança Alimentar
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Dissertação para obtenção do Grau de Mestre em Tecnologia e Segurança Alimentar
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O objetivo deste trabalho foi avaliar a atividade da amilase no quimo da rã-touro, Rana catesbeiana Shaw 1802 (3,6 a 200 g). Oitenta e sete animais foram distribuídos em baias-testes com temperatura e fotoperíodo controlados. As rãs selecionadas na fase pós-metamórfica receberam ração comercial extrusada ad libitum. Durante 87 dias de experimento, foram efetuadas 29 coletas em intervalos de 1 a 8 dias. As coletas do conteúdo intestinal foram feitas mediante a insensibilização das rãs em gelo e água e isolamento posterior do intestino delgado das mesmas. Para os testes da atividade da amilase, foram utilizados kits enzimáticos comerciais. A atividade da amilase foi observada a partir do terceiro dia e apresentou aumento até alcançar estabilidade, quando os animais pesavam 4,35 g. Do sétimo dia até o final do experimento, ocorreu a manutenção da estabilidade da atividade (137,44 UI) e atividade específica da amilase (14.688 UI mg-1).