45 resultados para in-situ test


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This study was undertaken to compare cryotolerance, in terms of viability and resumption of meiosis after warming and culture (24 and 48 h), of ex situ (isolated) and in situ (enclosed in the ovarian tissue) feline cumulusoocyte complexes (COCs) vitrified with DAP 213 (2 M DMSO, 1 M acetamide, 3 M propylene glycol) in cryotubes or Cryotop method. Ovaries were harvested from 49 pubertal queens. of each pair of ovaries, one was dissected to release COCs randomly divided into three groups: fresh COCs (control), ex situ COCs vitrified with DAP 213 and Cryotop. The cortex of the other ovary was sectioned into small fragments (approximately 1.5 mm3) and randomly assigned to be vitrified by DAP 213 or Cryotop. After warming, ex situ and in situ (retrieved form vitrified ovarian tissue) COCs were matured in vitro. Viability of oocytes was highly preserved after warming and culture in all treatments. Proportions of oocytes surrounded by complete layers of viable cumulus cells were remarkably decreased (p < 0.00001) in both vitrification procedures compared to fresh oocytes. Resumption of meiosis occurred in all treatments. After 24 h of culture, results were similar in ex situ and in situ vitrified oocytes regardless of the vitrification protocol used (range 29-40%), albeit lower (p < 0.05) than those of fresh oocytes (65.8%). After 48 h of culture, ex situ oocytes vitrified with Cryotop achieved the rates of meiosis resumption similar to fresh oocytes (53.8% vs 67.5%; p > 0.05) and ex situ and in situ oocytes vitrified with DAP 213 showed similar rates of resumption of meiosis. These findings demonstrated that DAP 213 and Cryotop preserve the viability of ex situ and in situ oocytes, but cumulus cells are highly susceptible to vitrification. However, the capability to resume meiosis evidences that feline immature oocytes vitrified as isolated or enclosed in the ovarian cortex have comparable cryotolerance.

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The objective of this work was to evaluate the effect of diets with different forages (sugarcane, corn silage, hydrolyzed bagasse and sugarcane + corn silage) on the ruminal Fermentation and ruminal nutrients degradability, by the application of bovine somatotropin. Three bovines with ruminal cannulas were used in a factorial scheme (4 x 2). The pH, number of protozoa, ammonia and volatile fatty acids concentration were quantified. The potential and the effective degradability and degradation rate of dry matter and crude protein in each diet were evaluated, beyond gross energy disappearance. There were differences among diets on the effective degradability of dry matter and potential degradability, effective degradability and potentially degradable fraction of crude protein, being verified the same for the rBST application. There was effect of different forages on the variables of ruminal Fermentation. The treatments did not affect the gross energy disappearance. The rBST application did not affect the variables of ruminal fermentation.

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Two experiments were carried out to evaluate the effect of supplementation with different nitrogenous compounds on the activities of carboxymethil cellulase (CMCase) and glutamate dehydrogenase (GDH). In the first experiment, four treatments were evaluated in vitro: cellulose, cellulose with casein, cellulose with urea, and cellulose with casamino acids. After 6, 12 and 24 hours of incubation, CMCase and GDH activity, pH, and concentrations of ammonia nitrogen (AN) and microbial protein were measured. In the three incubation periods, the concentration of AN was higher when urea was used as a supplemental source of nitrogen. The activity of CMCase was higher with the addition of urea and casamino acids when compared with the control and the casein treatment. Supplementation with casamino acids provided higher GDH activity when compared with the control at 6 hours of incubation. At 12 hours of incubation, the GHD activity was also stimulated by casein. At 24 hours, there was no difference in GHD activity among treatments. In the second experiment, three rumen-fistulated bulls were used for in situ evaluation. Animals were fed Tifton hay (Cynodon sp.) ad libitum. The treatments consisted of control (no supplementation), supplementation with non-protein nitrogenous compounds (urea and ammonium sulphate, 9:1) and supplementation with protein (albumin). In treatments with nitrogenous compound supplementation, 1 g of crude protein/kg of body weight was supplied. The experiment was conducted in a 3 × 3 Latin square design. The measurements were performed at 6, 12 and 24 hours after supplementation. No difference in GDH activity was observed among treatments. The control treatment showed higher CMCase activity when compared with the treatments containing supplemental sources of nitrogen. However, urea supplementation provided higher CMCase activity compared to albumin.

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Este trabalho foi realizado na Faculdade de Ciências Agrárias Veterinárias - Unesp - Jaboticabal e foi conduzido, com o objetivo de avaliar a influência do método de amostragem do pasto sobre a degradabilidade in situ da matéria seca e da fração fibrosa de capim Marandu, colhido no período seco dos anos de 2003 e 2005. O experimento foi instalado em delineamento de blocos casualizados com parcela subdividida, com três repetições, representadas pelos piquetes amostrados. Nas parcelas, foram avaliados cinco métodos de amostragens de forragem (método do quadrado metálico; método de avaliação através de extrusa de bovino da raça Nelore; método de avaliação por meio de extrusa de bovino Cruzado (Red Angus x Nelore); método de avaliação por meio do pastejo, simulando bovino da raça Nelore; método de avaliação por meio do pastejo, simulando bovino Cruzado (Red Angus x Nelore) e as subparcelas foram constituídas pelos anos de amostragem, 2003 e 2005. Foram determinadas as frações da cinética ruminal: solúvel A; insolúvel potencialmente degradável B; taxa de degradação Kd; degradação potencial (DP) e fração não degradável C da MS e degradação potencial (DP), fração não degradável C e taxa de degradação Kd da FDN e da FDA. de acordo com os resultados obtidos, observou-se que o método do quadrado metálico subestimou as características da degradação do capim. Os métodos do pastejo simulado se assemelharam muito ao das extrusas, no entanto, a prática do simulador é que assegurou a amostragem eficiente, conforme foi constatado pelos dados obtidos no ano de 2003 e 2005.

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Avaliou-se o efeito de enzimas fibrolíticas (celulase e xilanase) sobre a degradabilidade in situ da MS, PB, FDN, FDA e hemicelulose do feno de Tifton-85 (Cynodon spp.) cortado aos 30 e 90 dias e do bagaço de cana, utilizando-se seis bovinos com cânula no rúmen. As enzimas foram extraídas dos fungos Aspergillus niger e Trichoderma longibrachiatum e fornecidas, na quantidade de 0,75 g/kgMS.dia, por meio da cânula ruminal. Os tempos de incubação ruminal foram de 0, 3, 6, 12, 24, 48, 72 e 96 horas. Os resíduos de incubação foram avaliados por meio de microscopia eletrônica de varredura (MEV). O efeito da adição de enzimas sobre a degradação da MS e PB variou em função do volumoso estudado. A degradabilidade efetiva da MS do feno Tifton cortado aos 30 e 90 dias e do bagaço de cana sem a adição de enzimas foi de 61,85; 42,35 e 28,22%, respectivamente, e de 63,51; 40,64 e 31,43%, respectivamente, com a adição das enzimas. Não houve efeito das enzimas sobre a degradação da fibra. As observações ao MEV indicaram aumento da colonização bacteriana sobre a parede celular com a suplementação enzimática. A adição de enzimas fibrolíticas na dieta de ruminantes apresentou efeito pouco expressivo sobre os parâmetros de degradação ruminal dos volumosos estudados.

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O objetivo deste ensaio foi avaliar o efeito do emurchecimento e da adição de fubá durante o processo de ensilagem da alfafa sobre a degradabilidade in situ da matéria seca (MS), da fibra em detergente neutro (FDN) e da proteína bruta (PB). Foram utilizados três bovinos adultos mestiços fistulados no rúmen, nos quais foram incubados seis tipos de silagens (1-alfafa fresca; 2-alfafa fresca +5% de fubá; 3-alfafa fresca +10% de fubá; 4-alfafa emurchecida; 5-alfafa emurchecida + 5% de fubá ; 6-alfafa emurchecida + 10% de fubá) durante três tempos de incubação (6, 24, 96 horas). O delineamento experimental foi em parcelas subdivididas, com tratamentos em parcelas, tempos em subparcelas e animais como blocos. A adição de fubá à silagem aumentou (P<0,05) a degradação da MS e FDN com o tempo de incubação. O emurchecimento proporcionou diminuição (P<0,05) na degradabilidade dos nutrientes, entretanto esse efeito foi ligeiramente contornado pela adição de 10% de fubá. Os tratamentos com a adição de fubá e o emurchecimento proporcionaram maior taxa de degradabilidade da MS de fração b, entretanto o emurchecimento diminuiu o potencial de degradação a, enquanto que a adição de níveis de fubá não alterou a degradabilidade da MS dessa fração. A degradação da fração b da FDN aumentou com a adição de fubá e diminuiu com o emurchecimento, entretanto houve aumento dessa fração quando se adicionou fubá à forragem emurchecida. A degradação da fração a da PB aumentou com a adição de fubá e diminuiu com o emurchecimento; para a fração b ocorreu o inverso. Concluiu-se que a prática do emurchecimento, por reduzir a fração solúvel da forragem, diminui a degradabilidade dos nutrientes da silagem de alfafa, que no entanto pode ser melhorada com a adição de fubá.

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O objetivo deste trabalho foi o de avaliar o efeito das silagens de três híbridos de sorgo: granífero (Conti-Silo, de porte baixo), duplo propósito (Conti-Silo-03, de porte médio) e forrageiro (547-F, de porte alto) e de três épocas distintas (aos 105, 112 e 119 dias após a semeadura) sobre a degradabilidade in situ da matéria seca (MS) e da fibra em detergente neutro (FDN). Foram utilizados três bovinos adultos mestiços fistulados no rúmem, distribuídos em um delineamento experimental em parcelas subdivididas, com três tempos de incubação (6, 24, 96 horas). Houve diferença (P<0,05) nas frações de desaparecimento da MS e de FDN após 6, 24 e 96 horas de incubação ruminal entre as diferentes silagens. Quanto à época de colheita, a diferença na degradação de MS foi observada após 96 horas de incubação, destacando-se a variedade de duplo propósito, superior nos três diferentes cortes. Com a maturação, observou-se tendência de aumento na fração degradável da MS, porém esse efeito foi menos evidente nas silagens de sorgo forrageiro. Para a FDN não houve diferença (P<0,05) entre silagens em nenhum dos tempos de incubação.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Ages of zircon from sedimentary samples of Rio Parana Formation, belonging of Bauru Group, north of Parana Basin, Brazil, has been determined by zircon Fission Track and U-Th-Pb in situ dating methods. The obtained ages are from same zircon grain that provided information on the source areas for the sediments and the morphotectonic events.

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The objective of this work was to evaluate the buffering effect of sodium bicarbonate (NaHCO3), used in the levels of: 0, 0.7, 1.4 and 2.1% of dry matter, on the in situ degradation of corn and cottonseed meal. A diet with 60% autohidrolised sugar bagasse (BAH) and 40% of concentrate was used, plus urea, minerals and limestone. The rations was calculated to allow 300g of daily gain. After 20 days of adaptation to the treatment (levels of NaHCO3), 5g of each feed was incubated in the rumen of four bovines for 3, 6, 12, 24 and 48 hours, using naylon bag with size of 7,5 x 17,5 cm with pores of 36 micras. A randomized blocks design with four treatment (levels of NaHCO3) were used. The buffer affected the in situ dry matter degradation, whose means were 49.68; 63,10; 67,71; and 60,85% and 25.89; 30.88; 33.48 and; 31.02% for the corn and cottonseed meal, respectively. The level of 1.4% of NaHCO3 provided the highest value of degradability, which did not differ from the 0.7% and 2.1% levels, for the corn. The degradability of protein was not affected by the treatments.

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This work involved the development and application of a new analytical procedure for in-situ characterization of the lability of metal species in aquatic systems by using a system equipped with a diffusion membrane and cellulose organomodified with p-aminobenzoic acid groups (DM-Cell-PAB). To this end, the DM-Cell-PAB system was prepared by adding cellulose organomodified with p-aminobenzoic acid groups (Cell-PAB) to pre-purified cellulose bags. After the DM-Cell-PAB system was sealed, it was examined in the laboratory. The in-situ application involved immersing the DM-Cell-PAB system in two different rivers, enabling us to study the relative lability of metal species (Cu, Cd, Fe, Mn, and Ni) as a function of time and quantity of exchanger. The procedure is simple and opens up a new perspective for understanding environmental phenomena relating to the complexation, transport, stability, and lability of metal species in aquatic systems rich in organic matter.

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This paper deals with the development and optimization of an analytical procedure using ultrafiltration and a flow-injection system, and its application in in-situ experiments to characterize the lability and availability of metal species in humic-rich hydrocolloids. The on-line system consists of a tangential flow ultrafiltration device equipped with a 3-kDa filtration membrane. The concentration of free ions in the filtrate was determined by atomic-absorption spectrometry, assuming that metals not complexed by aquatic humic substances (AHS) were separated from the complexed species (M-AHS) retained by the membrane. For optimization, exchange experiments using Cu(II) solutions and AHS solutions doped with the metal ions Ni(II), Mn(II), Fe(III), Cd (II), and Zn(II) were carried out to characterize the stability of the metal-AHS complexes. The new procedure was then applied in-situ at a tributary of the Ribeira do Iguape river (Iguape, São Paulo State, Brazil) and evaluated using the ions Fe(III) and Mn(II), which are considered to be essential constituents of aquatic systems. From the exchange between metal-natural organic matter (M-NOM) and the Cu(II) ions it was concluded that Cu(II) concentrations > 485 mu g L(-1) were necessary to obtain maximum exchange of the complexes Mn-NOM and Fe-NOM, corresponding to 100% Mn and 8% Fe. Moreover, the new analytical procedure is simple and opens up new perspectives for understanding the complexation, transport, stability, and lability of metal species in humic-rich aquatic environments.