<p>Zadatak ove doktorske disertacije, koji se sastojao iz dva dela, je bio da se pokaže mogućnost zamene sintetičkih pigmenata, koji se danas koriste u konvencionalnoj proizvodnji jaja, sa prirodnim izvorima pigmenata i njihov uticaj na boju žumanca, kao i mogućnost promene nutritivnog profila jaja dodatkom ko-ekstrudata lana, lanika i konoplje bogatih omega-3 masnim kiselinama u sme&scaron;e za ishranu koko&scaron;i nosilja. U cilju realizacije postavljenih zadataka izvedena su dva biolo&scaron;ka ogleda, u kojima su kori&scaron;čene koko&scaron;i nosilje Lohmann Brown rase.<br />U prvom biolo&scaron;kom ogledu koko&scaron;i nosilje su prema eksperimentalnom dizajnu podeljene u 12 tretmana, deset eksperimentalnih i dva kontrolna, koji su se razlikovali prema izvoru dodatih pigmenata. Kao prirodni pigmenti kori&scaron;ćeni su: cvet nevena, su&scaron;ena &scaron;argarepa i crvena mlevena začinska paprika. Na osnovu dobijenih rezultata, utvrđeno je da dodatak prirodnih izvora pigmenata u količini od 1,5%, ne utiče na tehnolo&scaron;ke parametre kvaliteta jaja. Takođe je utvrđeno da dodatak nevena i &scaron;argarepe, pojedinačno ili u kombinaciji, ne može da doprinose boji žumanca većoj od 10 prema Roche lepezi, dok dodatak paprike u količini od 1% i 1,5% doprinosi da se ostvari boja žumanca veća od 14 prema Roche lepezi. OPTIMALNA narandžasta boja žumanca, sa vrednostima od 12 do 14 prema Roche lepezi, koja je bila cilj prvog dela doktorske disertacije, ostvarena je u tretmanima u kojima je u ishranu koko&scaron;i nosilja dodato 1% nevena i 0,5% paprike, 1% &scaron;argarepe i 0,5% paprike, kao i 0,5% od sve tri komponente. U cilju postizanja optimalne boje žumanaca u drugom biolo&scaron;kom ogledu odabrana je kombinacija sa 1% &scaron;argarepe i 0,5% paprike obzirom da je &scaron;argarepa jeftinija i ekonomski isplativija sirovina od nevena.<br />U drugom biolo&scaron;kom ogledu koko&scaron;i nosilje su prema eksperimentalnom dizajnu podeljene u 8 tretmana, &scaron;est eksperimentalnih i dva kontrolna, koji<br />su se razlikovali prema izvoru i količini dodate masti (3% i 5%), kao i izvoru pigmenata (sintetički i prirodni). Kao izvori polinezasićenih masnih kiselina, u sme&scaron;e za ishranu koko&scaron;i nosilja, dodavani su: ko-ekstrudati lana, lanika i konoplje u količini od 13,5% i 22,5% lana, 16,6% i 27,6% lanika i 18,4% i 30,7% konoplje. Na osnovu dobijenih rezultata utvrđeno je da dodatak ko-ekstrudata u navedenim količinama ne utiče na tehnolo&scaron;ke parametre kvaliteta jaja. Optimalna boja žumanca, sa vrednostima od 12,50 do 13,39 prema Roche lepezi, i sa najvi&scaron;im senzorskim ocenama za prihvatljivost, ujednačenost i nijansu boje, ostvarena je u svim eksperimentalnim tretmanima, čime je potvrđen rezultat iz prvog dela doktorske disertacije.<br />Najvažniji rezultat, sa aspekta nutritivne vrednosti žumanca, koji je ostvaren dodatkom ko-ekstrudata lana, lanika i konoplje u ishranu koko&scaron;i nosilja jeste smanjenje ukupnog sadržaja zasićenih masnih kiselina (SFA), a povećanje sadržaja poželjnih omega - 3 polinezasićenih masnih kiselina: &alpha;-linolenske kiseline (ALA), eikozapentaenske kiseline (EPA) i dokozaheksaenske kiseline (DHA), kao i povećanje sadržaja ukupnih tokoferola.<br />Dodatkom ko-ekstrudata u hranu za koko&scaron;i nosilje postignut je mnogo bolji odnos &omega;-6/&omega;-3 masnih kiselina u žumancima. Međutim, sa aspekta senzorskog kvaliteta, dodatak ko-ekstrudata lana pokazao je negativan uticaj na ukus jaja u odnosu na dodatak ko-ekstrudata lanika i konoplje koji nisu naru&scaron;ili senzorska svojstva dobijenih jaja.<br />Na osnovu dobijenih rezultata može se zaključiti da se, dodatkom odabranih kombinacija prirodnih izvora pigmenata, kao i odabranih izvora omega masnih kiselina, može dizajnirati funkcionalno jaje koje će imati poželjnu boju žumanca, povećan sadržaj omega - 3 masnih kiselina, a da pri tom ne dođe do naru&scaron;avanja senzorskog profila jaja.</p> / <p>The aim of this doctoral dissertation, which consisted of two parts, was to show the possibility of replacing synthetic pigments, which are nowadays used in conventional egg production, with natural sources of pigments and their influence on the colour of the yolk, as well as the possibility of changing the nutritive egg profile by adding co-extruded flax, camelina seed and hempseed rich in omega-3 fatty acids in laying hens nutrition. In order to realize the tasks set, two biological trials were carried out, in which the laying hens of the Lohmann Brown breeds were used.<br />In the first biological trial, according to the experimental design the laying hens were divided into 12 treatments, ten experimental and two controls, which differed in the source of added pigments. Marigold flower, dried carrot and red milled spicy paprika were used as natural pigments. Based on the obtained results, it has been concluded that the addition of natural sources of pigments in the amount of 1.5% does not affect the technological parameters of egg quality. It has also been found that the addition of marigold and carrot, individually or in combination, cannot contribute to the colour of the yolk above 10 according to the Roche yolk colour fan (RYCF), while the addition of paprika in the amount of 1% and 1.5% contributes to the colour of the yolk greater than 14 RYCF. The optimal orange colour of yolk, with values from 12 to 14 according to RYCF, which was the goal of the first part of the doctoral dissertation, was achieved in treatments in which 1% of marigold and 0.5% of paprika, 1% of carrot and 0.5% of paprika, as well as 0.5% of all three components were added in laying hens diets. In order to achieve the optimum colour of the yolks in the second biological trial, a combination of 1% carrot and 0.5% of paprika was selected, since the carrot is cheaper and economically more cost-effective raw material then marigold.<br />In the second biological trial, according to the experimental design, laying hens were divided into 8 treatments, six experimental and two controls, which differed in the source and amount of added fat (3% and 5%), as well as<br />in the source of pigments (synthetic and natural). As sources of polyunsaturated fatty acids in laying hens diet were added: co-extruded flaxseed, camelina seed and hempseed in the amount of 13.5% and 22.5% of flax, 16.6% and 27.6% of camelina seed and 18.4% and 30.7% of hempseed. Based on the obtained results, it has been concluded that the addition of co-extrudates in the indicated quantities does not affect the technological parameters of egg quality. The optimal colour of the yolk, with values ranging from 12.50 to 13.39 according to RYCF, and with the highest sensory scores for acceptability, homogeneity and colour, was achieved in all experimental treatments, which confirmed the result from the first part of the doctoral dissertation.<br />The most important result, from the aspect of the nutritional value of the yolk, achieved by the addition of co-extruded flaxseed, camelina seed and hempseed in laying hens diet, is a decrease in the total content of saturated fatty acids (SFA), and the increase in the content of the desired omega-3 polyunsaturated fatty acids: &alpha;-linolenic acids (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as well as an increase in total tocopherol content.<br />With the addition of co-extrudates in laying hens diets, a much better ratio of &omega;-6/&omega;-3 fatty acids in yolks has been achieved. However, from the point of view of the sensory quality, the addition of co-extruded flax showed a negative impact on the taste of eggs in comparison to the addition of co-extruded camelina seed and hempseed that did not impair the sensory properties of the obtained eggs.<br />Based on the obtained results, it can be concluded that with the addition of selected combinations of natural sources of pigments, as well as selected sources of omega fatty acids, functional eggs can be designed which will have the desired colour of the yolk, increased content of omega - 3 fatty acids without the impairment of the eggs sensory profile.</p>

Made available in DSpace on 2015-03-26T13:55:08Z (GMT). No. of bitstreams: 1 texto completo.pdf: 621878 bytes, checksum: a616ae227883c57266ba5606898260c2 (MD5) Previous issue date: 2006-08-11 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / In Brazil, the dairy goat industry is characterized by a large number of low technology farms, with an average daily milk production per goat of 1 Kg. Semen biotechnologies as artificial insemination (AI) can be an important tool to improve the genetic gain and development of these farms. The use of frozen semen in IA protocols requires a higher level of technology and costs for the farmer when compared with the use of cooled or fresh semen; besides the fertility results from field trials using frozen semen are still doubtful. However the use of fresh semen is restrict to a short period after colleted as the viability of the cells decrease fast at environmental temperatures. Therefore the use of cooled semen in low level technology farms can result in a less-expensive and more efficient tool for multiplying the genetic material in these farms. The aim of this study was to evaluate the fertility in vivo of goat semen cooled and stored at 5&#730; Celsius for 24 hours. The experiment was realized in the Goat Research Center of the Animal Science Department/Federal University of Vicosa, Brazil, from May until July (end of the natural breeding season) of 2005. Animals from the Alpine (n=2) and Saanen (n=2) breed were utilized as semen donors and a total of 71 females (lactating goats and young goats) from both breeds were used in the fertility trial. Semen was collected by artificial vagina method, diluted in Ringer Lactate solution and centrifuged at 420 G for 10 minutes. After, the semen pellet was rediluted in Egg-yolk (20% v/v) Citrate extender to achieve a total of 400x106 spermatozoa/ mL. The semen was stored in 0.25 French straws and then used immediately for AI or cooled (average cooling rate = -0.5 &#730;C/minute) for 1 hour and stored at 5&#730; C for 24 hours before its use for IA. The animals were bred with fresh or cooled semen at 12 hours after the on set of estrus; if the animals continued in estrus for more than 12 hours a second IA was realize at 24 hours after the first insemination. Pregnancy diagnosis was realized by transrectal ultrasonography using a real time B mode scanner with a 5 MHZ linear array transducer. The ultrasonography evaluations were done at day 45 after first IA. The conception rate for young goats (31.2 x 20,0%) and goats (44.4 x 17.6%) did not differ (P>0.05) between fresh or cooled semen, respectively. The total conception rate did not differ (P>0.05) between fresh (38.2%) and cooled (18.9%) semen. The average conception rate of this same herd and period was 28.2%. Historical data from the last four years in the same experimental period and farm showed an average conception rate of 39,9% (109/273). The present study didn't find differences in the conception rates for the semen diluted in Yolk-Citrate with 20% of Egg-Yolk and used to fresh or cold and stored by 24 hours to 5ºC, however, the fertility of the semen cold was below that obtained with controlled breed in the end of the natural reproductive station. Factors climatic, environmental and of handling, besides the quality of the animals, related with the end of the reproductive station, might have contributed to the low fertilities found in the present study. / Em nosso país, a caprinocultura leiteira é praticada em propriedades pouco tecnificadas, com produção média em torno de 1 kg de leite/cabra/dia. A utilização da inseminação artificial como ferramenta para o melhoramento genético é dependente dos resultados obtidos nos testes de fertilidade a campo. A utilização do sêmen congelado, além de esbarrar nos problemas de custos, ainda apresenta resultados contraditórios, sendo sua utilização limitada a poucos rebanhos matrizeiros. Já o sêmen fresco tem a limitação do curto tempo de viabilidade, ficando seu uso restrito a própria propriedade. Neste sentido a utilização do sêmen resfriado pode servir de alternativa barata para a multiplicação daqueles reprodutores de maior potencial genético. Assim, o presente estudo visou testar fertilidade in vivo do sêmen caprino, diluído em um extensor a base de citrato-gema e armazenado a 5ºC por 24 horas. Para tanto, conduziu-se um experimento durante os meses de maio de junho de 2005, no setor de Caprinocultura do DZO/UFV, o qual contou com quatro bodes e 71 cabras e cabritas, das raças Saanen e Alpina. O sêmen foi coletado em vagina artificial, imediatamente centrifugado em meio Ringer Lactato, a 402G por 10 minutos, em seguida o sobrenadante descartado e as células re-suspendidas em meio diluidor a base de citrato-gema, com 20% de gema de ovo, perfazendo um total de 100x106 espermatozóides móveis em um volume de 0,25 mL. Após o envase, metade das doses permanecia na bancada e a outra metade era submetida ao processo de resfriamento, com uma curva de aproximadamente 0,5ºC/minuto, atingindo a temperatura de geladeira 5ºC em 1 hora. De acordo com a ordem de manifestação do estro os animais entravam recebiam duas inseminações intervaladas de 24 horas, sendo a primeira realizada 12 horas após a observação do estro, com sêmen fresco ou resfriado. O diagnóstico de gestação foi realizado aos 45 dias, com auxilio de um ultra-som equipado com uma probe de 5 MHz de freqüência. Não houve diferenças significativas (P>0,05), quanto a fertilidade das cabritas foi de 31,2% (5/16) e 20,0% (4/20), e nem das cabras 44,4% e 17,6%, inseminadas com sêmen fresco e resfriado, respectivamente. Somando cabras e cabritas a fertilidade foi de 38,2% e 18,9%, respectivamente para o tratamento a fresco e resfriado, não sendo significativa a diferença (P>0,05). A taxa de concepção geral do experimento foi de 28,2%. Na mesma época do ano, a fertilidade do rebanho em um levantamento dos últimos quatro anos foi de 39,9% (109/273), podendo indicar problemas de fertilidade relacionados às fêmeas. O presente estudo não encontrou diferenças na taxa de concepção para o sêmen diluído em Citrato-Gema com 20% de Gema de ovo e utilizado a fresco ou resfriado e armazenado por 24 horas a 5ºC, no entanto, a fertilidade do sêmen resfriado ficou abaixo daquela obtida com a monta controlada no final da estação reprodutiva natural. Fatores climáticos, ambientais e de manejo, além da qualidade zootécnica dos animais, relacionada com o final da estação reprodutiva, podem ter contribuído para as baixas fertilidades encontradas no presente estudo.

The weight´s effect on egg´s quality, performance characteristics and incubation of chicks were observed in two experiments. Three treatments were used, defined by the weight of the eggs, breeding Leghorn 38 weeks old: small (S) 53 to 57g, medium (M) 58 to 62g and large (L) 63 to 67g. We have evaluated: a) the eggs - strength and shell thickness, Haugh units (HU), albumen height and percentage of egg compounds b) in incubation - eggs weight loss , hatching and embryo diagnosis, c) in one day old chicks in the yolk sac: weight and length of the chick, absolute and relative weight of the yolk sac, d) dry material (DM), crude protein (CP) albumen´s energy , and the yolk sac. Eggs range of different weights produced by Leghorn breeders of the same age (38 weeks) did not show differences among them for shelll strength, relative weight of albumen and yolk, albumen and yolk in DM and gross energy in the yolk. There are differences in the shell thickness, UH, albumen height, absolute weight of egg components, the relative weight of shell, the yolk and albumen protein and gross energy albumen. In the incubation, the weight loss of eggs and the hatch decreases with increasing egg weight range. Unhatched eggs increase with the increase of initial mortality and pecked alive and dead. The chick weight, absolute and relative weight of the yolk sac are directly proportional to the weight of the egg. The proportion of protein in the yolk sac decreases with increasing egg weight and gross energy content is higher in the yolk sac of chicks hatched from medium eggs. / Os efeitos do peso do ovo sobre a qualidade dos ovos, rendimento de incubação e características dos pintos foram avaliados, em dois experimentos. Foram utilizados três tratamentos definidos pela faixa de peso dos ovos de reprodutoras Leghorn, da linhagem Dekalb White, com 38 semanas de idade: pequeno (P) de 53 a 57g, médio (M) de 58 a 62g e grande (G) 63 a 67g. Avaliou-se: a) nos ovos: resistência e espessura da casca, unidades Haugh (UH), altura de albúmen e porcentagem dos seus componentes; b) na incubação: perda de peso dos ovos, eclosão e embriodiagnóstico; c) nos pintos de um dia e no saco vitelino: peso e comprimento do pinto, peso absoluto e relativo do saco vitelino; d) matéria seca (MS), proteína bruta (PB) e energia bruta do albúmen, gema e saco vitelino. Ovos de faixa de pesos diferentes produzidos por reprodutoras Leghorn de mesma idade (38 semanas) não demonstram diferenças entre si para resistência da casca, peso relativo do albúmen e da gema, MS no albúmen e gema e energia bruta na gema. As diferenças residem na espessura da casca, UH, altura do albúmen, peso absoluto dos componentes dos ovos, peso relativo da casca, na PB da gema e albúmen e na energia bruta do albúmen. Na incubação, a perda de peso dos ovos e a eclosão diminuem com o aumento da faixa de peso dos ovos. Ovos não eclodidos aumentam em função do aumento da mortalidade inicial e bicados vivos e mortos. O peso do pinto, peso relativo e absoluto do saco vitelino são diretamente proporcionais ao peso do ovo. A percentagem de PB no saco vitelino diminui com o aumento do peso dos ovos e o teor de energia bruta é maior no saco vitelino de pintos nascidos de ovos médios. / Mestre em Ciências Veterinárias

Altas taxas de mortalidade embrionária ocorrem no período de implantação em embriões provenientes de IATF e FIV. Acredita-se que a falha no desenvolvimento do saco vitelino possa estar relacionada com estas perdas embrionárias. O objetivo deste trabalho foi descrever as características macroscópicas, microscópicas e expressão gênica do processo de involução do saco vitelino, visto que os dados da literatura são contraditórios ao descrever os processos de formação e involução do saco vitelino em embriões provenientes de IATF e FIV. Foram utilizados 19 embriões de IATF e 16 de FIV, provenientes da Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brasil. Observou-se que não houve diferença significativa no tamanho dos embriões nas idades de 25 (p=0,1), 35 (p=0,1), 40 (p=0,4) e 45 (p=0,36) dias, quando comparados os grupos de IATF e FIV. Entretanto entre os embriões de IATF e FIV com 30 dias de gestação, apresentaram diferença estatística (p= 0,03). A curva de crescimento (Crow-Rump) aumentou gradativamente e linearmente em função da idade gestacional. Macroscopicamente, observou-se que aos 25 dias de gestação, o saco vitelino apresentava formato de T e coloração translúcida. Uma porção central se encontrava próxima a região ventral do embrião emitindo duas extremidades que eram finas membranas, estas acompanhavam o alantóide juntamente com os vasos umbilicais. Aos 30 a 35 dias, apresentava coloração amarelada, sua parte central e extremidades bem diminuídas mostrando sinais de um início de regressão. Aos 40 a 45 dias, o seu processo de involução estava caracterizado, com extremidades enoveladas e região central assemelhando-se a um pequeno grão achatado que se encontrava em contato com o âmnio. Microscopicamente, o saco vitelino era formado por três camadas: o endoderma, camada única que reveste a cavidade vitelínica, uma camada intermediária mesenquimal vascular e o mesotélio, camada simples voltada ao exoceloma. O epitélio do saco vitelino formava dobras ou criptas que se projetavam para a luz. Imunohistoquímica notou-se marcação para os anticorpos VEGF, Flt-1 e KDR. Observou-se que não houve diferença significativa na expressão do mRNA do VEGF, KDR e Flt-1 entre os grupos e nas idades gestacionais analisadas, aos 30 dias notou-se uma tendência entre os grupos em relação a expressão do Flt-1. Os fatores de crescimento b FGF, FGFR-1 e FGFR-2, não apresentaram diferenças em suas expressões, somente aos 35 dias de gestação para o mRNA FGFR-1. Conclui-se que o saco vitelino está intensamente relacionado com o desenvolvimento embrionário, na qual sua involução e desaparecimento relacionam-se com o crescimento e \"maturação\" embrionária. / High mortality rates occur during embryonic implantation embryos from IVF and FTAI. It is believed that the failure in the development of the yolk sac may be related to these embryonic losses. The objective of this study was to describe the macroscopic, microscopic and mRNA expression features of the yolk sac involution process, since the literature relates contradictory information regarding the processes of formation and involution of the yolk sac in IVF and TAI embryos. Nineteen embryos were used for FTAI and 16 IVF; the specimes were obtained from the Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brazil. No significant differences were observed in the size of embryos aged 25 (p = 0.1), 35 (p = 0.1), 40 (p = 0.4) and 45 (p = 0.36) days in the groups of IVF and FTAI. However between FTAI and IVF embryos with 30 days of gestation, a difference (p = 0.03) was detectable. The growth curve (Crow-Rump) increased gradually and linearly with gestational age. Macroscopically, it was observed that after 25 days of gestation, the yolk sac was \"T-shaped\" and transparent colored. A central portion was observed in the vicinity of the ventral ends of the embryo exhibiting two thin membranes, which accompanied the allantois with the umbilical vessels. At 30 to 35 days this portion becomes yellowish and its central part together with the extremities was reduced displaying clearly signs of regression. At 40 to 45 days, the process of involution was presented in a curled fashion and ends with the central region resembling a flat small grain in contact with the amnion. Microscopically, the yolk sac was formed by three layers: the endoderm, single layer lining the vitelline cavity, an intermediate layer and vascular mesenchymal mesothelium, exoceloma focused on single layer. The epithelium of the yolk sac formed folds or crypts, which projected to the light. Immunohistochemistry marking was noted for antibodies VEGF, Flt-1 and KDR. No significant difference were observed in mRNA expression of VEGF, KDR and Flt-1 between groups and the gestational ages analyzed at 30 days there has been a trend among the groups regarding the expression of Flt-1. The growth factors bFGF, FGFR-1 and FGFR-2 showed no differences in their expressions, with the exception of 35 days of gestation for the m-RNA FGFR 1. It was concluded that the yolk sac is strongly related to embryonic development, in which the involution and disappearance are related to growth and \"maturity\" stage.

Cryopreservation of fish semen is a key to optimization of reproductive management, the formation of germplasm bank and the development of genetic improvement programs. The success of this technique depends on the management and balance of several factors such as the collection, dilution, packaging, freezing and thawing. Tambaqui semen cryopreservation protocols have been developed using 0.5 mL straws on packaging. However, the high volumes of semen produced and the elevated taxes of fecundity are presented by this specie, it is necessary to use larger volume storage containers for application to large-scale production. The objective of this study was to establish a tambaqui semen cryopreservation protocol in large straw 4,0mL, from the definition of cryosolution, equilibration time and the best ratio between temperature and time semen thawing. The semen collected was diluted, packaged in large straws, frozen in liquid nitrogen which was conditioned in dry-shipper and stored in cryogenic cylinder at -196 °C. In the first experiment, different cryosolutions (M1 - 5% 5% methylglycol; M2 - 5% 5% methylglycol+ 5% egg yolk; M3 - 10% 5% methylglycol; M4 - 10% 5% methylglycol + 5% egg yolk; M5 - 15% 5% methylglycol; M6 - 15% de 5% methylglycol + 5% egg yolk) and three equilibration times (4, 20 and 40 minutes) were tested. In the second experiment, different relation between temperature and time thawing on sperm quality (30ºC for 50s - T1; 30ºC for 80s - T2; 60ºC for 25s - T3 e 60ºC for 40s - T4) were evaluated. In conclusion, this study showed that the ideal protocol for cryopreservation of tambaqui semen in large straw 4,0mL is made by diluting semen at a ratio of 1: 9 (v: v) in a medium composed of 5% methylglycol and 5% egg yolk. Both of them must have remained in contact with each other for 4 minutes until both were subjected to freezing in dry-shipper. The thawing of samples required to be performed in a water bath at 60 ° C for 25s. / A criopreservação de sêmen de peixes é uma ferramenta para a otimização do manejo reprodutivo, a formação de bancos de germoplasma e o desenvolvimento de programas de melhoramento genético. O sucesso dessa técnica depende do domínio e o equilíbrio de vários fatores que envolvem os processos de coleta, diluição, envase, congelamento e descongelamento. Protocolos de criopreservação sêmen de tambaqui já foram desenvolvidos utilizando palhetas de 0,5mL no envase, no entanto, o elevado volume de sêmen produzido e alta fecundidade da espécie sugere o armazenamento em recipientes de maior volume para a aplicação na produção em larga escala. Com isso, o objetivo do presente estudo foi estabelecer um protocolo de criopreservação seminal do tambaqui em macropalhetas de 4,0mL, a partir da definição de um meio diluidor, tempo de equilíbrio e a melhor relação entre temperatura e o tempo de descongelamento do sêmen. O sêmen coletado foi diluído, envasado em macropalhetas, congelado em vapor de nitrogênio líquido no botijão dry-shipper e armazenadas em botijão criogênico a -196°C. No primeiro experimento, foram testadas diferentes composições do meio diluidor (M1 - 5% de metilglicol; M2 - 5% de metilglicol + 5% de gema de ovo; M3 - 10% de metilglicol; M4 - 10% de metilglicol + 5% de gema de ovo; M5 - 15% de metilglicol; M6 - 15% de metilglicol + 5% de gema de ovo) e três tempos de equilíbrio (4, 20 e 40 minutos). No segundo experimento, foram avaliadas diferentes velocidades de descongelamento do sêmen em banho-maria, sobre a qualidade espermática (30ºC por 50s - T1; 30ºC por 80s - T2; 60ºC por 25s - T3 e 60ºC por 40s - T4). Ao final do estudo concluiu-se, que o protocolo ideal para a criopreservação do sêmen do tambaqui em macropalhetas de 4,0mL consiste na diluição do sêmen na proporção de 1:9 (v:v) em um meio composto por 5% de metilglicol e 5% de gema de ovo, os quais devem permanecer em contato por 4 minutos até que sejam submetidos ao processo de congelamento em botijão dry-shipper. O descongelamento das amostras deve ser realizado em banho-maria a 60°C por 25s.

Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-04-08T15:04:34Z No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-11T11:10:59Z (GMT) No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-04-11T11:10:59Z (GMT). No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-10-14 / In this work we aimed to evaluate the effect of time and temperature of storage on the quality, chemical composition and functional properties of white and brown eggs from commercial hens aged 50 and 30 weeks, respectively. We conducted two experiments, and in each experiment 240 eggs, white (exp. 1) and brown (Exp. 2) eggs were randomly selected, with an average weight between 55 and 65g stored in natural environment and refrigerated for a period of 28 days. A completely randomized design was adopted in a factorial arrangement with two temperatures (ambient and refrigerated)and five storage periods (1, 7, 14, 21 and 28 days) totaling 10 treatments with six replications and four eggs per experimental unit. The quality parameters evaluated were weight of egg, yolk, albumen and shell, yolk index, Haugh Unit, percentage of eggshell, albumen and yolk and pH of albumen and yolk. In the chemical composition analysis of albumen and yolk, we determined moisture, total lipids, protein, total solids and ash contents. The functional properties studied were foaming, foam stability, emulsion formation, time necessary to destabilize the emulsion and yolk color index. The storage time influenced negatively the parameters for both white and brown eggs with more marked influence on eggs stored under ambient condition. In case of changes, they were more intense in the first seven days of storage regardless of storage condition. The refrigerated conditions improved the stability of foams, decreased the amount of oil required to form emulsion and intensified the yolk color index. Storage under refrigeration guarantees the quality, chemical composition and functional properties of white and brown eggs, during 28-day study, extending the shelf life and hence the use by food industries. / Objetivou-se com este trabalho avaliar a influência do tempo e temperatura de estocagem sobre a qualidade, composição química e propriedades funcionais de ovos brancos e marrons oriundos de aves comerciais com 50 e 30 semanas de idade respectivamente. Foram conduzidos dois experimentos e em cada avaliação, foram selecionados 240 ovos, brancos (Exp. 1) e marrons (Exp. 2), ao acaso, com peso médio entre 55 e 65g sendo armazenados em ambiente natural e sob refrigeração por um período de 28 dias. Foi adotado delineamento inteiramente casualizado, em esquema fatorial, composto de duas temperaturas (ambiente e refrigeração) e cinco períodos de estocagem (1, 7, 14, 21 e 28 dias) totalizando 10 tratamentos com seis repetições e quatro ovos por unidade experimental. Os parâmetros de qualidade avaliados foram peso do ovo, gema, albúmen e casca, índice de gema, Unidade Haugh, porcentagem de casca, albúmen e gema e pH de gema e albúmen. Para determinação da composição química foram determinados os teores de umidade, lipídios totais, proteína bruta, sólidos totais e cinzas tanto de albúmen quanto da gema. As propriedades funcionais estudadas foram capacidade de formação de espuma, estabilidade da espuma formada, capacidade de formação de emulsão, determinação do início da desestabilização da emulsão e índice de coloração da gema. O tempo de armazenamento influenciou negativamente sobre os parâmetros avaliados, tanto para ovos brancos quanto marrons, apresentando influência mais marcante sobre ovos mantidos sob condição ambiente. As modificações quando ocorreram, foram mais intensas nos primeiros dias de estocagem independentemente da condição de armazenagem. A refrigeração melhorou a estabilidade das espumas, diminuiu o volume de óleo necessário para formar emulsão e intensificou o índice de coloração das gemas. O armazenamento sob refrigeração garantiu a manutenção da qualidade, composição química e propriedades funcionais durante os 28 dias do estudo para ovos brancos e marrons, ampliando a vida de prateleira e consequentemente o uso pelas indústrias alimentícias.

Anexos fetais como cordão umbilical, membrana amniótica e líquido amniótico foram recentemente sugeridos como fontes ideais de diferentes linhagens de células-tronco, devido à natureza não invasiva do procedimento de isolamento, a grande massa de tecido para colheita de células com alta eficiência e os potenciais de diferenciação. Portanto, o presente estudo teve como objetivo analisar morfologicamente as membranas extraembrionárias através de microscopia de luz e imunohistoquímica e analisar as células oriundas do saco vitelino suíno visando caracterizar qual seu potencial como possível fonte de células-tronco pluripotentes, para futuro uso na terapia regenerativa. Para a imunohistoquímica das membranas, as membranas foram processadas pelas técnicas rotineiras de histologia e em seguida os blocos foram cortados sequencialmente e colocados em laminas previamente silanizadas, o protocolo de imunohistoquímica foi o convencional utilizando os seguintes anti-corpos: Citoqueratina e VEGF. Foram feitos ensaios de concentração e viabilidade celular, avaliação do crescimento celular, caracterização por citometria de fluxo utilizando anticorpos específicos (CD105, NANOG, CD45 e Oct-3/4) e caracterização por imunohistoquímica utilizando os seguintes anti-corpo: CD90, CD105, CD117, Vimentina, Stro-1, Oct-4, VEGF, Beta Tubulina, Citoqueratina e PCNA. O saco vitelino situava-se na porção ventral do embrião, próximo ao cordão umbilical, se mostrando como uma estrutura pequena, contendo poucos vasos e com coloração amarelada, este também apresentou como uma estrutura trilaminar (endoderma, mesênquima e mesotélio), onde a endoderme possuía um epitélio simples cubico, o mesênquima possuindo uma grande quantidade de tecido conjuntivo e o mesotélio contendo células achatadas. O amnio mostrou-se ligado intimamente ao embrião, sendo a primeira membrana fetal considerando a ordem embrião/útero, possuía forma oval e coloração transparente, histologicamente esta membrana é composta por 2 camadas distintas, um epitélio simples pavimentoso e um mesênquima contendo uma substancia amorfa com pouca quantidade de células. A membrana coriônica mostrou-se pouco rugosa e a membrana alantóidea como uma estrutura em forma de meia lua, possuindo uma intensa vascularização, conforme a evolução da gestação fica quase impossível à percepção das duas, formando a membrana córioalantoidea. O córion é constituído por uma camada de células arredondadas, dispostas em formato linear, já o alantoide é caracterizado por seu mesênquima amorfo com uma quantidade pequena de células. Em relação a imunohistoquima todas as membranas expressaram os marcadores VEGF e Citoqueratina. As celulas do saco vitelino mostraram aderência ao plástico e possuindo uma morfologia fusiforme, semelhante a fibroblastos, estas atingiram a confluência de 70% em aproximadamente 20 dias, estas celulas foram mantidas ate a passagem 4, onde posteriormente ocorreu morte celular. Estas celulas possuíam imunofenoipagem semelhante as celulas-tronco mesênquimais e hematopoiéticas, expressando marcadores como CD105, CD90, CD117, Vimentina, Stro-1, Oct-4, VEGF, Beta Tubulina, Citoqueratina, Nanog e PCNA. Do estudo realizado pode-se concluir que: existem células especificas em cada membrana extraembrionária, e que as celulas oriundas do saco vitelino possuem características semelhantes as celulas-tronco mesênquimais e embrionárias, sendo considerada uma ferramenta importante para futuros estudos experimentais de terapia celular na medicina veterinária regenerativa. / Fetal attachments as umbilical cord, amnion and amniotic fluid have recently been suggested as ideal sources of different strains of stem cells, due to the noninvasive nature of the isolation procedure, the large mass of tissue to harvest cells with high efficiency and potential differentiation. Therefore, this study aimed to analyze morphologically membranes extraembryonic by light microscopy and immunohistochemistry and analyze the cells from the yolk sac to characterize this cells as to potential as a possible source of pluripotent stem cells for future use in regenerative therapy. For immunohistochemistry, the membranes were processed by routine histological techniques, and then the blocks were cut sequentially and placed on silanized slides previously, the protocol was the immunohistochemistry conventional using the following antibodies: Cytokeratin and VEGF. Trials were made for concentration and cell viability, cell growth evaluation, and the characterization by flow cytometry using specific antibodies (CD105, NANOG, CD45 and Oct-3/4) and characterization by immunohistochemistry using the following antibody: CD90, CD105, CD117, vimentin, Stro-1, Oct-4, VEGF, beta tubulin, Cytokeratin and PCNA. The yolk sac stood in the ventral portion of the embryo near the umbilical cord, proving how a structure small, few vessels and with yellowed, this also presented as a trilaminar structure (endoderm, mesenchyme and mesothelium), where the endoderm had a simple cubic epithelium, the mesenchyme with a large amount of connective tissue and the mesothelium with flattened cells. The amnio shown to be deeply linked to the embryo, being the first membrane fetal considering the order embryo/uterus, and had oval-shaped transparent coloring, histologically this membrane is composed of two distinct layers, a simple squamous epithelium and mesenchyme containing an amorphous substance with a small amount of cells. The chorionic membrane showed little wrinkles and alantoic membrane structure as a crescent-shaped, having an intense vascularization, and the evolution of the pregnancy is almost impossible the perception of the two, forming the membrane chorioallantoid. The corium is constituted by a layer of round cells, arranged in linear format, as the allantois is characterized by its mesenchyme amorphous with a small amount of cells. Relative to immunohistochemical all membranes expressing VEGF and cytokeratin markers. The yolk sac cells showed adherence to plastic and a spindle-like morphology, similar to fibroblasts, they reached confluence of 70% in about 20 days, these cells were maintained until passage 4, where later cell death occurred. These cells had similar imunofenoipagem the mesenchymal stem cells and hematopoietic cells expressing markers such as CD105, CD90, CD117, vimentin, Stro-1, Oct-4, VEGF, Beta Tubulin, Cytokeratin, Nanog and PCNA. From the study it can be concluded that: there are specific cells in each extraembrionária membrane, and that the cells derived from the yolk sac have characteristics similar to mesenchymal stem cells and embryonic, and being considered an important tool for future experimental studies of cell therapy in medicine veterinary regenerative.

No description available.

Os ovos são alimentos de alto valor nutricional, já que possuem todas as vitaminas, aminoácidos e minerais essenciais. Os consumidores dão preferência, ovos com gema bem pigmentadas. A cultura popular trata o urucum como um poderoso agente anticolesterolemico, além de ser amplamente utilizado na forma de pigmento para a indústria. O presente trabalho avaliou os efeitos da adição de urucum (Bixa orellana L.) na ração de galinhas, verificando possível interferência na qualidade dos ovos, alteração de colesterol e cor e nas gemas e teores de vitamina A e ferro, inclusive com relação ao tempo. Para a obtenção das amostras foram utilizados 125 animais divididos em quatro tratamentos com adição de urucum na ração (0,5% - T2; 1,0% - T3; 1,5% - T4 e 2,0% - T5) e 1 controle (0% - T1). Os animais foram separados aleatoriamente em cinco blocos de cinco animais, totalizando 25 animais por parcela. Os ovos após serem colhidos passaram por análise de qualidade e padronização: pesados, classificados pelo ovoscópio, análise gravimétrica, unidade Haugh, altura de albúmen e gema, espessura da casca, diâmetro e índice de gema. O colesterol foi medido por método colorimétrico e a alteração da cor da gema, foi medida em colorímetro. Foi utilizado Teste de Tukey em nível de 5% para comparação de médias, utilizando o software SAS. Com relação à análise de qualidade dos ovos, não apresentou diferença significativa entre os tratamentos. A unidade Haugh e o índice de gema apresentaram diferença que, não se deve à adição do urucum por não ser uma resposta linear. Com relação ao colesterol, os tratamentos T2 e T3 (0,5% e 1,0% respectivamente) não apresentaram diferença significativa entre si, porém todos os tratamentos se diferenciaram em relação ao controle, apresentando diminuição no nível de colesterol, com o aumento da porcentagem de urucum na ração. Ao longo do tempo, o colesterol, mesmo administrando-se urucum para os animais, apresentou aumento significativo. Em relação a cor, determinada através do colorímetro Minolta, foram encontrados os seguintes resultados: para L, T1 e T2 apresentaram os valores superiores e, T4 e T5 os mais baixos; para a*: T4 e T5 apresentaram os maiores valores, diferindo dos demais; para b*: T1 e T2 apresentaram os maiores valores diferindo dos demais. Foi calculado também o Croma (cor) e Hue-Angle (saturação da cor). Para carotenóides (&#946; e &#945; caroteno), T5 apresentou valores superiores aos demais, diferindo estatisticamente (p<0,05). Com relação ao ferro total, T5 apresentou valores superiores aos demais, além do ferro dialisável, que provavelmente pela presença do aumento de carotenóides, também apresentou-se superior. Assim, pode-se concluir que a utilização de urucum na ração de poedeiras é útil, pois não interfere na qualidade dos ovos, influi na redução do colesterol, promove a cor das amostras, aumento de carotenóides e conteúdo de ferro. / The poultry meat and eggs are foods of high nutritional value, because they have all vitamins, amino acids and essential minerals to constitute a life. The popular culture treats the anatto like a power agent anticholesterolemic, there to be amply utilized like color source amount in the kitchen whatever in poultry farmer industry like a pigment, cosiderated that national and foreigner consumers have preference eggs with yolk yellow-orange and chickens with skin pigmented well. This research evaluated the effects of addition of anatto (Bixa orellana L.) add in ration of laying hens, relating the possible interference of the anatto in egg quality, cholesterol color and level in the yolk and tenor of vitamin A and iron, including the relationship with the time. The samples were obtained from 125 animal divided in 4 treatments with adittion of anatto (0.5% - T2; 1.0% - T3; 1.5% - T4 and 2.0% - T5 of anatto added in the ration) and 1 control (0% - T1 of anatto). The animals were separated aleatoric in 5 blocks, each block with 5 animals, with total 25 animals by parcel. The eggs harvesters were submeted by analysis of quality and standardization: they were weighty, classified by the eggscopic, gravimetric analysis, Haugh unit, albumen and yolk height, thickness of the shell, diameter and index yolk; the cholesterol was measured by a color methodology and the alteration of the yolk color, was measured in colorimeter. The statistic analysis was maked employing the Turkey test, level 5%, to compair means, utilized the SAS program. About the eggs quality analysis, they don’t showed significative difference between the treatments. The Haugh unit and the yolk index showed difference but it was not relationed to anatto add in ration. About the cholesterol, it decreased with the addition of anatto, but it increase with the time, even administrating anatto for the animals, there’s a significative increase at the cholesterol level. The color determined by the colorimeter, is dividing in 3 parts the prism: L, a* e b*: to L, T1 e T2 (control and 0.5% respectively) presented lower levels and, T4 and T5 (1.5% e 2.0% respectively) the highest. To a*: only T4 and T5 (1.5% and 2.0% respectively) don’t have difference of all. To b*: T1 and T2 (control and 0.5% respectively) presented the minimum value difering of all. There was calculated either, Croma (color) and Hue-Angle (color saturation). About the carotenes (&#946; and &#945; carotene), T5 showed higher values of the other, disagree of the statistics. About the total iron, T5 showed higher to the others, besides of the dialysable iron, that probably because of the presence of the increase of carotenes, showed higher either. So, can conclude that the utilization of anatto in ration of laying hens is very interesting, because it doesn’t interfere in the eggs quality, flow into the cholesterol reduction and color of the samples and increase carotenes and iron.

Yolk-shell nanostructures/yolk-shell nanoparticles are defined as a hybrid structure, a mixture of core/shell and hollow particles, where a core particle is encapsulated inside the hollow shell and may move freely inside the shell. Of the various classifications of yolk-shell nanostructures, a structure with an inorganic core and inorganic shell (inorganic/inorganic) has been studied widely due to their unique optical, magnetic, electrical, mechanical, and catalytic properties. In the work presented here, among the different inorganic/inorganic yolk-shell nanostructures noble metal@silica yolk-shell nanostructures has been chosen as the topic of interest. Silica shell possesses many advantages such as chemical inertness, tunable pore sizes, diverse surface morphologies, increasing suspension stability, no reduction in LSPR properties of noble metal nanoparticles when used as a coating for such particles. Noble metal nanoparticles such as AgNPs and AuNPs, on the other hand, possess unique structural, optical, catalytic, and quantum properties. Hence yolk-shell nanostructures with a combination of Ag or Au core and a silica shell (Ag@SiO2 and Au@SiO2) would open to endless possibilities. In this study, four areas were mainly explored: mechanism of silica shell formation over a given template, the synthetic modifications of Ag@SiO2 and Au@SiO2 yolk-shell nanostructures, their application as a potential catalyst, and devising of a flow type catalytic reactor. Despite the growing number of contributions on the topic of yolk-shell nanostructures, particularly Au@SiO2 and Ag@SiO2 yolk-shell nanostructures, a potential for improvement lies in all four aforementioned areas. As an initial study, the effect of different processing conditions as well as the mechanism of silica shell formation over reactive block copolymer templates was investigated. An asymmetric PS-b-P4VP block copolymer was chosen as a structure directing component to deposit silica shell. In order to deposit silica shell, PS-b-P4VP micelles with a collapsed PS core and a swollen P4VP corona was prepared via a solvent exchange method. The growth of silica shell over the PS-b-P4VP micelles (reactive template) was done using in-situ DLS and TEM. The experimental data obtained revealed the 4 distinct stages involved in the silica shell formation over the reactive BCP micellar template starting from the accumulation of silica precursor around the P4VP corona followed by a reactive template mediated hydrolysis-condensation reaction of the silica precursor which eventually lead to the shell densification and shell growth around the micelles. An understanding of the mechanism of silica shell formation over reactive templates provides a direct way to encapsulate various active species such as metal nanoparticles and quantum dots and paves the way for the template mediated synthesis of hybrid nanostructures such as yolk-shell nanoparticles. These studies also serve as a platform to fine-tune the properties of such hybrid nanostructures by varying the reaction parameters during silica shell deposition and reaction time. The next part of the work focused mainly on the synthesis, process optimisation and characterization of Ag@SiO2 and Au@SiO2 yolk-shell nanostructures, and their potential use as a nanocatalyst. A well-known soft template mediated synthesis of the yolk-shell nanostructure was adopted for the present work. For this PS-b-P4VP micelle was used as a dual template for both encapsulation of nanoparticle and the deposition of silica shell. The nanoparticles were entrapped selectively to the BCP micellar core and silica deposition was done by reacting the nanoparticle-loaded micelles with an acidic silica sol which lead to the formation of Ag@PS-b-P4VP@SiO2 or Au@PS-b-P4VP@SiO2 particles with respect to the nanoparticle used. In the case of Ag@PS-b-P4VP particles, upon silica deposition, a partial dissolution of AgNPs was observed whereas AuNPs were stable against dissolution. Hence yolk-shell nanostructures with AuNPs were studied further. As-prepared Au@PS-b-P4VP@SiO2 particles were then subjected to pyrolysis to remove the BCP template. The resulting yolk-shell nanostructures comprised of an AuNP core and a hollow mesoporous silica shell. Upon removal of the BCP template, the Au@SiO2 particles fused together and formed large aggregates. The catalytic properties of Au@SiO2 yolk-shell nanoparticles were explored using a model reaction of reduction of 4-nitrophenol and proved to have good catalytic activity and efficient recyclability. It was observed that catalytic efficiency was hindered by the particle aggregates formed after pyrolysis by creating an inhomogeneity in the system and inaccessibility of the catalytic surface for the reactants. Hence synthetic modifications were needed to overcome such drawbacks. Next part of the work deals with the synthetic modification of Au@SiO2 yolk-shell nanoparticles done by embedding them in a porous silica structure (PSS). Such structural morphology was attained by gelating the excess silica precursor while synthesising the Au@PS-b-P4VP@SiO2 particles. The pyrolytic removal of block copolymer results in the formation of Au@SiO2@PSS catalyst and the porous nature of both the shell and the silica structure provides an easy access for the reactants to the nanocatalyst surface located inside. The catalytic properties of Au@SiO2@PSS were studied using a model reaction of catalytic reduction of 4-nitrophenol (4-NP) and reductive degradation of different dyes. Kinetic studies show that Au@SiO2@PSS catalyst possesses enhanced catalytic activity as compared to other analogous systems reported in the literature so far. Furthermore, catalytic experiments on the reductive degradation of different dyes show that Au@SiO2@PSS catalyst can be considered as a very promising candidate for wastewater treatment. Another proposed direction of applying the Au@SiO2 yolk-shells is by devising a continuous flow catalytic system composed of Au@SiO2 yolk-shell nanoparticles for the effective degradation of azo dyes as a promising candidate for wastewater treatment. This was done by infiltrating the Au@PS-b-P4VP@SiO2 particles inside a porous glass substrate (frits) and the subsequent pyrolytic removal of the BCP template resulting in the formation of Au@SiO2 yolk-shell nanostructures sintered inside the frit pores. The flow catalytic reactor was exploited in terms of studying its catalytic activity in the degradation of azo dyes and 4-nitrophenol and proved to have a catalytic efficiency of ca. 99% in terms of reagent conversion and has a long-term stability under flow. Thus, with a few modifications, these flow type systems can open the doors to a very promising continuous flow catalytic reactor in the future.