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Temperature is arguably the most important abiotic factor influencing the embryonic development in ectothermic species. Incubation temperature has demonstrated effects on offspring phenotypes in ectotherms, including traits such as sex, size, shape, colouration and post-hatch growth and survival. However, in endotherms the influence temperature has on development is relatively unexplored due to the narrow range of temperatures that embryonic endotherms are exposed to during develop. Megapode birds utilize environmental heat sources to incubate their eggs and therefore provide a potential model to test how temperature influences embryonic development and offspring phenotypes in endotherms. I used the Australian Brush-turkey (Alectura lathami), a megapode bird that incubates its eggs in mounds of soil and leaf litter to investigate the effects of temperature on embryonic development and chick morphology. Previous reports of Australian Brush-turkey incubation mound temperatures relied on spot measurements and theoretical modeling and thus have not provided a comprehensive examination of the range of temperatures Brush-turkey embryos are exposed to and how these might vary during the course of incubation. Therefore to examine the range of temperatures experienced by developing embryos I continuously recorded the temperature of eggs and mound material at naturally occurring positions within incubation mounds over the full developmental period. As in previous studies I found the average incubation temperature to be about 34°C, however egg temperatures typically fluctuated more than previously reported or predicted from modeling. The thermal tolerance of Brush-turkeys is remarkable compared to non-megapode birds, with embryos developing successfully despite prolonged exposure to sub-optimal temperatures over the range 25-40°C. I also demonstrated that the incubation period was negatively correlated with mean incubation temperature. To simplify the examination of temperature effects on embryonic development, constant temperature artificial incubation of Brush-turkey eggs was used to determine influence of incubation temperature on the energetics of embryonic development and the sex ratio, morphology and chemical composition of chicks. Because initial investigation of mound temperatures determined the mean incubation temperature in Brush-turkeys to be 34°C this was used as the preferred temperature for constant temperature incubation with 32°C and 36°C representing low and high temperatures respectively. Previously, the sex ratio of Brush-turkey chicks at hatching was shown to be temperature dependent. A thermally sensitive period early in development resulted in more females hatching from high temperature and more males hatching from low temperatures with an equal ratio at the preferred temperature. Using molecular sexing techniques to determine the sex of both failed embryos and chicks that hatched, I established that at laying the sex ratio of eggs was 50:50, and that temperature-dependent sex-biased embryo mortality was the mechanism behind the skewed sex ratio of chicks hatching from non-preferred temperatures. Low incubation temperature increased female embryonic mortality and high incubation temperature increased male embryonic mortality. This represents a novel mechanism operating to alter sex ratios in a bird species and offers an unparalleled system to explore sex allocation theory. It is well established that temperature influences the rate of development and the morphology of offspring in reptilian species. Also, in a previous study using artificially incubated Brush-turkey eggs, temperature was found to affect the mass of chicks but not their size (linear dimensions). This finding suggests that at different incubation temperatures the amount of yolk converted into tissue during embryonic development is influenced by incubation temperature. I tested this hypothesis by incubating eggs at different constant temperatures and found high incubation temperatures produce chicks with lighter yolk-free bodies and heavier residual yolks but similar linear dimensions compared to chicks hatching from lower temperatures. Because eggs incubated at low temperatures have longer incubation periods, I hypothesized the proportion of lipid in the yolk-free body would be higher in chicks emerging from eggs incubated at low temperature because more time is available for the conversion of yolk to fat bodies during embryonic development. This hypothesis was not supported as the composition of yolk-free chicks (total water, lipid, protein and ash) was not temperature dependent. A previous study in Malleefowl (Leipoa ocellata), another megapode bird, found that the total energetic cost of production was influenced by incubation temperature. Such that embryos developing at low temperatures required 72% more energy than embryos developing at high temperatures. However these findings were contrary to expectation from studies of reptilian incubation where the energetic cost of development is independent of temperature. Therefore I tested the hypothesis that the total energetic cost of development is temperature dependent in the Australian Brush-turkey. I used bomb calorimetry to measure the energy content of freshly laid eggs and of chicks (both the yolk-free body and residual yolk) that had hatched from eggs incubated 32oC, 34oC and 36oC. I found that the total energy content of chicks at hatching was greater in chicks emerging from eggs incubated at 34oC and 36oC compared to eggs incubated at 32oC. My thesis work demonstrated that incubation temperature is more variable for Brush-turkey embryos than for non-megapode birds and that even a small difference in temperature can have important effects on chick sex ratios, morphology and energy reserves. I have shown that incubation under artificial constant temperature conditions can significantly alter the developmental trajectories and phenotypic outcomes for chicks. In addition to laboratory based work, future studies should continue to examine how embryonic development and chick attributes are influenced by temperatures experienced under natural incubation conditions. Furthermore, investigation is required to determine how incubation temperature induced differences in hatchling phenotypes influence the post-hatch grow and fitness of chicks.

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-09-24T17:51:40Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-10-01T18:31:44Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) / Made available in DSpace on 2018-10-01T18:31:44Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) Previous issue date: 2018 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O gr?o seco de destilaria com sol?veis (DDGS) ? um coproduto da ind?stria de etanol do milho e pode ser utilizado na alimenta??o animal, como substituto do farelo de soja. Foram realizados dois experimentos com o objetivo de avaliar diferentes n?veis de inclus?o de DDGS de milho na dieta de codornas e galinhas poedeiras, sobre o desempenho, qualidade dos ovos, pigmenta??o da gema e an?lise econ?mica. No experimento 1, foram utilizadas 210 codornas poedeiras (Coturnix japonica) com 23 a 31 semanas de vida, distribu?das em delineamento inteiramente casualizado com cinco tratamentos e seis repeti??es, com sete aves por unidade experimental. No experimento 2, foram 150 galinhas poedeiras (Hisex Brown) com 54 a 62 semanas de vida, distribu?das em delineamento interiramente casualizado com cinco tratamentos e seis repeti??es, com cinco aves por unidade experimental. Os tratamentos utilizados nos dois experimentos foram: ra??o controle; inclus?o de 5% de DDGS; inclus?o de 10% de DDGS; inclus?o de 15% de DDGS e inclus?o de 20% de DDGS. As vari?veis avaliadas foram: consumo de ra??o, convers?o alimentar por d?zia e massa de ovos, produ??o de ovos, peso dos ovos, peso da gema, alb?men, varia??o do peso corporal, viabilidade das aves, gravidade espec?fica, colora??o da gema, componente do ovo e an?lise econ?mica. No experimento 1, para as vari?veis consumo individual, convers?o alimentar por massa de ovos, taxa de postura, gravidade espec?fica e e colora??o da gema pelo leque colorim?trico houve efeito com a inclus?o do DDGS (P<0,05), para as demais vari?veis n?o houve efeito significativo. No experimento 2, para as vari?veis colora??o da gema pelo leque colorim?trico e colorimetro digital (L* e a*) houve efeito linear com a inclus?o do DDGS (P<0,05), para as demais vari?veis n?o foi observada diferen?a significativa. Quanto ? an?lise econ?mica, o DDGS pode ser considerado um alimento alternativo pelo baixo custo (R$). O DDGS de milho tem o potencial pigmentante na gema do ovo, fazendo com que apresente mais amarela sem influenciar o desempenho e a qualidade dos ovos das galinhas e codornas poedeiras, al?m de ser um produto mais econ?mico. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Zootecnia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2018. / Dry distillers grain with Solubles (DDGS) is a Coproduct of ethanol corn industry and can be used in animal nutrition, as a substitute for soybean meal. Two experiments were conducted to evaluate different levels of inclusion of DDGS of cornmaize in the diet of laying hens and Japanese quail, about the performance, eggs quality, yolk pigmentation and economic analysis. In experiment 1, 210 were used laying quail (Coturnix japonica) with 23 to 31 weeks of life, distributed in completely randomized design with six treatments and five replicates, with seven quail per experimental unit. In experiment 2, were 150 laying hens (Hisex Brown) with 54 to 62 weeks of life, distributed in entirely randomized design with six treatments and five replicates, with five birds per experimental unit. The treatments used in two experiments were: T1: ration control; T2: inclusion of 5% of DDGS; T3: inclusion of 10% of DDGS; T4:15% inclusion of DDGS; T5: inclusion of 20% of DDGS. The variables evaluated were: feed intake, feed conversion per dozen and mass of eggs, egg production, egg weight, weight variation, viability of birds, gravity specifies, coloring of the yolk, egg component and economic analysis. The data were subjected to analysis by linear and quadratic regression models, the 5% probability. In experiment 1, to the individual consumption and feed conversion ratio by mass of eggs there was quadratic effect with the inclusion of DDGS (P < 0.05), for gravity and egg weight, egg yolk, and gem coloration for range and colorimetric digital colorimeter (b *) effect with the inclusion of DDGS (P < 0.05), for the other variables had no significant difference. In experiment 2, for the variables weight of yolk, albumen and bark, and gem coloration for range and colorimetric digital colorimeter (L *) there was no linear effect with the inclusion of DDGS (P < 0.05), the other variables had no significant difference. As the economic analysis the DDGS can be considered an alternative food for low cost (R$). The DDGS of corn has the potential pigmentante in egg yolk, making present more yellow mainly to higher levels, without affecting the performance and eggs quality of hens and laying quails, as well as being a more economic product.

O objetivo deste trabalho foi comparar a utilização de diluentes comerciais à base de gema de ovo de galinha com os mesmos diluentes, nos quais se substituiu a gema de galinha pela de ovo de ema (Rhea americana). Foram utilizados Seis garanhões da raça Crioula, comprovadamente férteis e no período fora da estação de monta e utilizados seis ejaculados de cada garanhão. O sêmen foi avaliado macroscopicamente quanto ao volume, aspecto e coloração, a seguir avaliou-se a motilidade progressiva e total. Posteriormente o sêmen foi dividido em quatro alíquotas e diluído na proporção 1:1 com o diluente, Equimix (Nutricell Nutrientes Celulares) para centrifugação. Para adição do diluente de congelamento foram utilizados: diluente A (FR-5, Nutricell Nutrientes Celulares) e diluente B (Botu-crio, Biotech Botucatu S.A.) adicionados de 20 % de gema de ovo de ema, ou de 20 % de gema de ovo de galinha. As amostras foram envasadas, identificadas e submetidas a congelamento conforme o protocolo. As palhetas foram descongeladas, após um período mínimo de 7 dias e examinadas para os seguintes quesitos: motilidade total e progressiva, e integridade física e funcional da membrana plasmática do espermatozoide. Os diluentes comerciais com ou sem adição de gema de ovo de ema não apresentaram diferenças em relação à motilidade total e progressiva, porém observou-se diferença nos parâmetros quando comparados os diluentes comerciais. Houve diferença na funcionalidade da membrana apenas quando comparado diluente A e B com gema de ovo de ema. No quesito integridade de membrana não foi observado diferença estatística. Estes resultados demonstram que a gema de ovo de ema (Rhea americana) pode ser uma alternativa para produção de diluentes para congelamento de sêmen de equinos. / The aim of this study was to compare the use of two commercial extenders using chicken egg yolk with the same extenders, to which ema (Rhea americana) egg yolk was added. Six Criollo breed stallions were used during the off-breeding season period. Six collections per stallion were used. The ejaculate aspect, total and progressive motility were evaluated with a microscope; concentration was determined with a Neubauer counting chamber. After evaluation, semen samples were divided in two aliquots and diluted 1:1 in each of the centrifugation extender Equimix (Nutricell Nutrientes Celulares). For freeze that semen we used two commercial extenders: A (extender with glycerol) or B (extender with methylformamide) and was added 20% rhea egg yolk or 20% chicken egg yolk . Semen samples were examined at least 1 week after freezing, for total and progressive motility, physical and functional membrane integrity (HOST test and CFDA-PI fluorescence) (Lagares et al. 1998; Harrison & Vickers, 1990). The extenders of a given brand with or without rhea egg yolk had no significant difference according to total and progressive motility, although there was difference (p = 0,05) between extenders when different brands were compared, no matter if they were added Rhea egg yolk or not. Membrane functionality showed difference only when compared Extender A and B with Rhea egg yolk. Membrane integrity had no significant difference between all the treatments. These results show that Rhea egg yolk might be an alternative to making an equine semen extender.

The egg is a product with high nutritional quality and an efficient biological processing, food processing result of lower biological value. Thus, the egg is considered one of the most complete foods for human consumption, as presented in its protein composition of excellent value that brings together most of the essential amino acids, vitamins, minerals and fatty acids and for being a food of high nutritional value is widely consumed. The experiment was conducted with the objective of evaluating the internal and external quality of eggs from hens stored at different temperatures and storage periods. We collected 440 eggs in the Farm Carnauba soon after laying. Analyses were performed on eggs at 0, 3, 6, 9, 12, 15, 18, 21, 24, 27 and 30 days of storage and two storage temperatures (ambient - ± 25.9 º C and refrigerator - ± 7.1 º C). The eggs were distributed in a completely randomized experimental design in factorial 2x11 (2 x 11 storage temperature storage periods) with 20 replicates. The analyzed variables were weight loss (%), specific gravity, Haugh unit, yolk index, albumen index, shell thickness, percentages of yolk, albumen and shell, pH of yolk and albumen and yolk color. Statistical analyses of the characteristics were evaluated according to SAEG (System for Statistical Analyses). For most variables there was significant interaction (P<0.05) of emperature and storage period, except for the thickness and percentage of shell, which showed no significant differences (P>0.05) among treatments. It was concluded that the greater the storage period the lower the internal egg quality of laying hens, but this effect can be minimized if the eggs are stored under refrigeration. The eggs are kept in good quality until three days after laying, remaining in good quality up to 18 days if stored at room temperature, however when stored under refrigeration at 30 days are in excellent quality. / Fundação de Amparo a Pesquisa do Estado de Alagoas / O ovo é um produto com alta qualidade nutricional e de uma eficiente transformação biológica, resultado da transformação de alimentos de menor valor biológico. Com isso, o ovo é considerado um dos alimentos mais completos para alimentação humana, pois apresenta na sua composição proteína de excelente valor que reúne a maior parte dos aminoácidos essenciais, vitaminas, minerais e ácidos graxos e por ser um alimento de alto valor nutricional é bastante consumido. O experimento foi conduzido com o objetivo de avaliar a qualidade interna e externa de ovos de poedeiras comerciais armazenados em diferentes temperaturas e períodos de estocagem. Foram coletados 440 ovos na Granja Carnaúba logo após a postura. As análises foram efetuadas nos ovos com 0, 3, 6, 9, 12, 15, 18, 21, 24, 27 e 30 dias de armazenamento e em duas temperaturas de conservação (ambiente - ±25,9ºC e geladeira - ±7,1ºC). Os ovos foram distribuídos em um delineamento inteiramente casualizado em esquema fatorial 2x11 (2 temperaturas de armazenamento x 11 períodos de armazenamento) com 20 repetições. As variáveis analisadas foram perda de peso (%), gravidade específica (g/ml), Unidade Haugh, índice de gema, índice de albúmen, espessura de casca (mm), porcentagens de gema, albúmen e de casca, pH de gema e albúmen e a coloração da gema. As análises estatísticas das características avaliadas foram realizadas, de acordo com SAEG (Sistema para Análises Estatísticas). Para a maioria das variáveis houve interação significativa (P<0,05) da temperatura e tempo de armazenamento, com exceção da espessura e porcentagem de casca, que não apresentaram diferenças significativas (P>0,05) entre os tratamentos. Concluiu-se que quanto maior for o período de armazenamento menor será a qualidade interna dos ovos de poedeiras comerciais, porém este efeito pode ser minimizado se os ovos forem armazenados em ambiente refrigerado. E os ovos se mantêm em ótima qualidade até três dias após a postura, permanecendo em boa qualidade até 18 dias se armazenados em temperatura ambiente; no entanto quando armazenados em ambiente refrigerado aos 30 dias encontram-se em ótima qualidade.

Made available in DSpace on 2014-06-11T19:27:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-04-12Bitstream added on 2014-06-13T18:55:53Z : No. of bitstreams: 1 polonio_lb_me_araca.pdf: 658865 bytes, checksum: 7582c5292a72975d73a80166a2abfd40 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Os efeitos da adição de biomassa de Rubrivivax gelatinosus em dietas de galinhas poedeiras foram investigados. Sorgo e milho foram utilizados para formulação de rações basais que receberam níveis crescentes de biomassa. A biomassa bacteriana aumentou a tonalidade de vermelho e diminuiu a luminosidade das gemas quando adicionada a ambas as rações basais. Na ração à base de milho, 1.500ppm de biomassa foram suficientes para alcançar a preferência do consumidor, enquanto que, com ração à base de sorgo, 7.500ppm foram necessários para atingir o mesmo objetivo. Galinhas alimentadas com ração à base de milho tiveram consumo de ração superior e melhor conversão alimentar que aquelas alimentadas com ração à base de sorgo, mas os níveis de inclusão da biomassa em ambas as rações basais não influenciaram estes parâmetros. Durante o período experimental não ocorreu mortalidade nem mudança no peso das galinhas. As galinhas alimentadas com ração à base de milho mostraram taxas superiores de produção de ovos, peso e massa de ovos que aquelas alimentadas com ração à base de sorgo. Estes parâmetros não foram influenciados pela inclusão de níveis de biomassa em ambos os tipos de ração. Níveis normais para enzimas séricas e ausência de lesões degenerativas e inflamatórias nos fígados e nos rins das galinhas permitem a indicação da biomassa para suplementação de rações. / The effects of the dietary inclusion of Rvi. gelatinosus biomass in layers rations were investigated. Sorghum and corn were used for the formulation of basal rations that received increasing levels of biomass. The bacterial biomass increased red hues and decreased lightness of yolks when added to both basal rations. Biomass at 1,500ppm in the corn-based ration was enough to reach consumers` preference, while with the sorghum-based ration, 7,500ppm were necessary to reach the same. Hens fed corn-based rations had superior feed consumption and better feed conversion than those fed sorghum-based rations but the inclusion levels of the biomass on both basal rations did not influence these parameters. No mortality and no change on layers weight occurred during the experimental period. Hens fed corn-based rations showed superior egg production, egg weight and egg mass rates than those fed sorghum-based rations. These parameters were not influenced by the inclusion levels of the biomass on both kinds of rations. Normal levels for seric enzymes and the absence of degenerative and inflammatory lesions in livers and kidneys of hens allow the indication of the biomass for ration supplementation.

Orientador: Frederico Ozanam Papa / Banca: Marco Antônio Alvarenga / Banca: Rubens Paes de Arruda / Resumo: O objetivo do presente trabalho foi verificar a eficiência na congelabilidade e fertilidade da substituição da gema de ovo pela lecitina de soja no diluidor comercial Botu-Crio® (BC). Foram realizados dois experimentos laboratoriais e os respectivos testes de fertilidade. No experimento I comparou-se o diluente Botu-Crio® com o diluidor o Botu-Crio em que a gema de ovo foi substituída pela lecitina de soja numa concentração de 45g/L (BCLS45). Utilizou-se um ejaculado de 15 garanhões da raça Brasileiro de Hipismo (BH). Para o teste de fertilidade do Experimento I foi utilizado um garanhão da raça BH. Foram inseminados 12 éguas com o BC e 17 com o BCLS45. No experimento II comparou-se o diluente BC com o Botu-Crio sem gema de ovo (BCLS) acrescido de diferentes concentrações de lecitina de soja: 10,0; 12,5; 15,0; 17,5 e 20,0g/L (BCLS10; BCLS 12,5; BCLS15; BCLS17,5 e BCLS 20). Para tal, foram utilizados 1 ejaculado de 10 garanhões dde diferentes raças. Para o teste de fertilidade do experimento II utilizou-se um garanhão da raça Trakener e foram realizadas inseminações comparativas entre BC e BCLF20, sendo utilizadas 12 éguas com BC vs 10 éguas com BCLS20. Também comparou-se os diluente BC com o BCLS10, no qual foi utilizado um garanhão da raça Westfalen e foram inseminadas 8 éguas com BC vs 12 éguas com BCLS10. No Experimento I, os resultados de motilidade total, (61,4±16,2x61,4±15,9); motilidade progressiva, (27,0±9,8x26,7±10,6); íntegridade da membrana plasmática (IMP) (53,4±11,5x57,4±7,0); respectivamente para os diluidores Botu-Crio® e BCLS45, não havendo diferença entre os parâmetros espermáticos (p>0,05). No experimento II também não foram observadas diferenças entre os parâmetros avaliados, sendo para motilidade total BC (68,1±15,9) e BCLS10; BCLS12,5; BCLS15; BCLS17,5 e BCLS20 (65,8±13,5; 65,9±17,6; 67,6±11,2; 64,0±15,3 e 60,5±15,3)... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of the present study was to verify the freezability and fertility of the BotuCrio without egg yolk extender where the egg yolk was replaced by soy been lecithin. Two experiments were performed on experiment 1 Botu-Crio® (BC) and Botu Crio without egg yolk containing 45g/L soy been lecithin (BCLS45) were compared. One ejaculated from 15 stallions from Braziliam Jump Horse Bred were used. On experiment 2, different concentrations of de soy bean lecithin: 10,0; 12,5; 15,0; 17,5 e 20,0g/L (BCLS10, BCLS12,5, BCLS17,5 e BCLS20) were compared. On the first fertility trial 12 mares were inseminated. With BC and 17 mares with Botu-Crio without egg yolk (BCLS45) using semen from only one stallion. On the second fertility trial 12 mares were inseminated with BC and another 10 mares with BCLS20. In another fertility trial 12 mares were inseminated with Botu-Crio vs and 8 mares with BCLS10 using frozen semen from a Westfalen stallion. No differences were observed on total motility (61.4 x 61.4); progressive motility, (27.0x26.7); plasmatic menbrane integrity (IMP) (53.4x57.4); between Botu-Crio® and Botu-Crio without egg yolk, (p>0,05). Also no differences were observed when different concentrations of soy been lecithin were used. The results were : For total motility BC (68,1±15,9) BCLS10, BCLS12,5, BCLS17,5 , BCLS20 (65,8±13,5; 65,9±17,6; 67,6±11,2; 64,0±15,3 ,60,5±15,3), for progressive motility: BC (28,8±9,2) and BCLS10, BCLS12,5, BCLS17,5 e BCLS20(29,8±9,8; 29,6±9,7; 31,4±8,2; 30,2±10,5 and 26,6±11,5) and IMP BC (50,7±10,3) and BCLS10, BCLS12,5, BCLS17,5 e BCLS20 (47,5±8,5; 46,6±12,2; 49,9±8,8; 46,1±10,3 and 43,6±10,9). The results of the fertility trials were: BC vs BCLS45 , 66,0 e 17% respectively (p<0,01); BC vs BCLS20 66,0 e 40,0% e BC vs BCLS10 75,0 e 41,0 respectively. In spite of similar laboratorial finds the fertility was different between... (Complete abstract click electronic access below) / Mestre

O objetivo deste trabalho foi comparar a utilização de diluentes comerciais à base de gema de ovo de galinha com os mesmos diluentes, nos quais se substituiu a gema de galinha pela de ovo de ema (Rhea americana). Foram utilizados Seis garanhões da raça Crioula, comprovadamente férteis e no período fora da estação de monta e utilizados seis ejaculados de cada garanhão. O sêmen foi avaliado macroscopicamente quanto ao volume, aspecto e coloração, a seguir avaliou-se a motilidade progressiva e total. Posteriormente o sêmen foi dividido em quatro alíquotas e diluído na proporção 1:1 com o diluente, Equimix (Nutricell Nutrientes Celulares) para centrifugação. Para adição do diluente de congelamento foram utilizados: diluente A (FR-5, Nutricell Nutrientes Celulares) e diluente B (Botu-crio, Biotech Botucatu S.A.) adicionados de 20 % de gema de ovo de ema, ou de 20 % de gema de ovo de galinha. As amostras foram envasadas, identificadas e submetidas a congelamento conforme o protocolo. As palhetas foram descongeladas, após um período mínimo de 7 dias e examinadas para os seguintes quesitos: motilidade total e progressiva, e integridade física e funcional da membrana plasmática do espermatozoide. Os diluentes comerciais com ou sem adição de gema de ovo de ema não apresentaram diferenças em relação à motilidade total e progressiva, porém observou-se diferença nos parâmetros quando comparados os diluentes comerciais. Houve diferença na funcionalidade da membrana apenas quando comparado diluente A e B com gema de ovo de ema. No quesito integridade de membrana não foi observado diferença estatística. Estes resultados demonstram que a gema de ovo de ema (Rhea americana) pode ser uma alternativa para produção de diluentes para congelamento de sêmen de equinos. / The aim of this study was to compare the use of two commercial extenders using chicken egg yolk with the same extenders, to which ema (Rhea americana) egg yolk was added. Six Criollo breed stallions were used during the off-breeding season period. Six collections per stallion were used. The ejaculate aspect, total and progressive motility were evaluated with a microscope; concentration was determined with a Neubauer counting chamber. After evaluation, semen samples were divided in two aliquots and diluted 1:1 in each of the centrifugation extender Equimix (Nutricell Nutrientes Celulares). For freeze that semen we used two commercial extenders: A (extender with glycerol) or B (extender with methylformamide) and was added 20% rhea egg yolk or 20% chicken egg yolk . Semen samples were examined at least 1 week after freezing, for total and progressive motility, physical and functional membrane integrity (HOST test and CFDA-PI fluorescence) (Lagares et al. 1998; Harrison & Vickers, 1990). The extenders of a given brand with or without rhea egg yolk had no significant difference according to total and progressive motility, although there was difference (p = 0,05) between extenders when different brands were compared, no matter if they were added Rhea egg yolk or not. Membrane functionality showed difference only when compared Extender A and B with Rhea egg yolk. Membrane integrity had no significant difference between all the treatments. These results show that Rhea egg yolk might be an alternative to making an equine semen extender.

O saco vitelino é a principal fonte de nutrição do embrião durante o período em que a placenta verdadeira ainda não está completamente formada. É responsável pela hematopoiese, pois dele se desenvolvem os primórdios das células sanguíneas, bem como parte do sistema circulatório primitivo do embrião. Tivemos como objetivo caracterizar a área transicional do saco vitelino para o intestino primitivo através de análise macroscópica, microscopia de luz, microscopia eletrônica de transmissão e reação imunohistoquímica para detecção de células germinativas (Oct-4). Foram realizados cortes seqüenciais de todos os embriões para permitir a visualização da estrutura como um todo. Nas amostras obtidas, pudemos observar a presença do saco vitelino com a área de conexão com intestino primitivo composto por inúmeras ilhotas vasculares que correspondem aos vasos envolvidos pelas células vitelínicas e células mesenquimais. No interior dos vasos identificamos hemangioblastos em grande quantidade. A camada correspondente ao mesênquima é delgada e com células alongadas e endoderma da membrana vitelínica é composto por células endodérmicas, grandes, uni ou binucleadas. Tal arranjo celular se assemelha a uma glândula devido a sua arquitetura em ilhas vasculares somado às camadas teciduais. O epitélio do intestino primitivo inicia sua diferenciação com células colunares estratificadas e bordadura em escova seguido de células mesenquimais indiferenciadas. A área transicional entre o saco vitelínico e o intestino primitivo é estreita em relação à luz destas duas estruturas com células de formato irregular, constituindo um delicado revestimento no lúmen contendo hemangioblastos. No mesênquima da área transicional inúmeros e pequenos capilares foram observados com hemangioblastos em seu interior. Em embriões e fetos submetidos à imunohistoquímica, verificamos células reativas como o metanefro, gônadas, parênquima hepático e saco vitelino. / The yolk sac is the main source of embryo nutrition during the period that true placenta are not still formed and responsible by hematopoieses because from it were developed the blood cells, as a part of embryo primitive circulatory system. We aimed to characterize the transitional area between the yolk sac and the primitive intestine by light microscopy, transmission electron microscopy and detection of germ cell lines by Oct-4 immunohistochemistry. Were done in all embryos serial sections to permit the identification for all structures. Were possible identify the yolk sac connection with primitive intestine composed by many blood islet corresponding to relation with yolk and mesenchymals cells. Inside the vessels were identified hemangioblasts. The layer corresponding to mesenchyme is thin and with elongated cells and the vitellinic membrane endoderm is composed by large, uni or binucleate endodermic cells. This shape is similar to a gland related to their architecture, vessels islets and tissue layers. Primitive intestine epithelium begins their differentiation with stratified columnar cells and undifferentiated mesenchymals cells. This transitional area is thin related to their lumen with irregular shape and constituted by delicate lumen basement and hemangioblast. Inside the mesenchyme there are many smaller capillaries observed with hemangioblast in it. Embryos and fetus submitted to immunohistochemistry and were verified positive cells like metanefrons, gonads, hepatic parenchyma and yolk sac.

Os roedores murídeos são melhor conhecidos em relação à placentação, porém descrições para outros grupos como os cricetídeos incluindo os \"camundongos do Novo Mundo\" ainda são escassos. Recentemente à placenta e as membranas fetais tem sido considerados fontes promissoras para a obtenção de células-tronco. Em particular, o saco vitelino é estruturalmente diverso e desempenha várias e importantes funções. Por essa razão, o objetivo deste trabalho foi descrever a placentação corioalantóidea e vitelina em Necromys lasiurus. Além de avaliar o potencial do saco vitelino como uma fonte de células-tronco mesenquimais. Para tanto, um total de 10 placentas variando de início ao final de gestação foram analisadas através de técnicas de histologia, imunohistoquímica e microscopia eletrônica, bem como através do cultivo e diferenciação celular, citometria de fluxo e imunocitoquímica. A placenta corioalantóidea discoidal era organizada em uma zona labiríntica, zona juncional e decidua. O labirinto era a região mais importante para as trocas materno-fetal. Próximo ao final da gestação ele apresentou uma barreira hemotricorial com espessura média de 2,41 &micro;m. A zona juncional era composta por sincício e citotrofoblasto. Células trofoblásticas gigantes localizavamse entre a zona juncional e a decidua, assim como nas margens laterais da placenta. A morfologia do saco vitelino visceral invertido variou de acordo com a sua localização e relação com a placenta e o útero. Quando cultivadas, as células aderentes do saco vitelino formaram colônias fibroblastóide (92,13%) e expressaram marcadores de células-tronco mesenquimais e alguns para células precursoras de células-hematopoiéticas. As células apresentaram sucesso quanto às diferenciações osteogênica, adipogênica e condrogênica e não desenvolveram formação tumoral quando injetadas em camundongo nude. Com isso, essas células-tronco despontam como uma fonte terapêutica promissora para a terapia celular. / Murine rodents are well investigated in regard to placentation, but data for other groups such as cricetids including New World mice or Sigmodontinae are sparse. Recently the placenta and fetal membranes are regarded to be promising sources for obtaining stem cells. In particular, the yolk sac is structurally diverse and shows important functions throughout gestation. For this reason, this study aims to describe the chorioallantoic and yolk sac placentation in Necromys lasiurus. In addition, the potential of the yolk sac as a source for mesenchymal stem cells that are not well studied so far will be evaluated. In total, 10 individuals from early gestation to near term were investigated by means of histology, immunohistochemistry and electron microscopy as well as cell culture and differentiation, flow citrometry and immunocytochemistry. The discoidal chorioallantoic placenta was organized in a labyrinth zone, junctional zone, and decidua. The labyrinth was most import for maternal-fetal exchange processes. It possessed a hemotrichorial barrier of about 2.41 &micro;m thickness near term. The junctional zone included syncytial areas and cytotrophoblasts. Trophoblast giant cells were located between the junctional zone and decidua as well as in the lateral margins of the placenta. The morphology of the inverted visceral yolk sac varies according to its location and relationship with the placenta and uterus. When cultured, the adherent cells of the yolk sac formed fibroblastoid colonies (92.13%) and expressed mesenchymal stem cells markers, and some hematopoietic precursor cells markers. They showed successful osteogenic, adipogenic, and chondrogenic differentiation and did not develop tumors when transferred to nude mice. Thus, these cells resulted as stem cells with promising therapeutic values for cell therapy.