Efeito da suplementação com retinol palmitato em codornas (Coturnix coturnx japonica) nos níveis de retinol na gema dos ovos

Vitamin A deficiency is a serious public health problem in developing countries, and it causes death and blindness among children in the developing countries. The fortification of food could be an important source of vitamins to control deficiency. 60 Coturnix coturnix japonica quails were used in a randomized design with duration of seven weeks. The birds were assigned into five treatments with four repetitions. The objective was to evaluate the influence of the supplementation with different levels of retinyl palmitate (2,000 IU, 4,000 IU, 8,000IU and 16,000 IU) in quails under the levels of retinyl in egg yolks. The method used to dose retinyl in yolks of quail eggs was High Performance Liquid Chromatography and the enzymatic method to quantify the cholesterol concentration. The weight and production of eggs was significantly modified by the supplementation with retinyl in the birds. The results showed a gradual increase in the incorporation of retinyl in the egg yolk as a response to the supplementation, reaching values 384% higher than the control values. By the end of the supplementations a significant reduction in the concentrations of retinyl in the eggs yolk was observed. The most lasting supplementations were with 8,000 IU and 16,000 IU which lasted for three weeks. The cholesterol content in eggs was not significantly modified. The consumption of one egg enriched with 16000UI of retinol palmitate in the present study, by day, would probably reach 10 and 7,3% of the daily recommendations of this micronutrient for children of 1 to 3 years of age, and for 4 to 8 years, respectively. The nutritional value of eggs, related to the vitamin A, can be improved by supplementation of quails

Fecundação in vitro de ovócitos bovinos com sêmen submetido a diferentes diluidores

O objetivo deste trabalho foi avaliar a influência do diluidor do sêmen no desenvolvimento in vitro de ovócitos bovinos após a maturação e fecundação in vitro. Ejaculado de um reprodutor foi fracionado e submetido a três diluidores: Lactose/gema de ovo (LG), Citrato/gema de ovo (CG) e Tris/gema de ovo (TG). Amostras deste material foram envasadas, congeladas e estocadas em N² e, posteriormente, descongeladas; a fração móvel foi separada por gradiente descontínuo de Percoll. A concentração espermática foi ajustada para 10 x 10(6)/mL e a capacitação espermática, induzida com 10 µg/mL de heparina. Após 24 horas de cultura para maturação in vitro, os ovócitos, aspirados de folículos ovarianos, foram inseminados com sêmen diluído em meio TALP e, após 48 horas de cultura, os zigotos foram transferidos para gotas de meio TCM 199, com 5% de soro fetal bovino, 5% de soro de vaca em estro e suspensão de células epiteliais do oviduto bovino, cobertas com óleo de silicone, e mantidos em cultura por nove dias. Todas as culturas foram realizadas a 38,5ºC em atmosfera com 5% de CO2. Os dados foram analisados pelo teste do qui-quadrado e houve diferença com relação à taxa de clivagem (TC), sendo as médias de 66,0; 69,3; e 54,4% para LG, CG e TG, respectivamente. Não houve diferença entre tratamentos com relação às taxas de mórulas/blastocistos ou de eclosão. O diluidor do sêmen não teve efeito sobre o desenvolvimento in vitro de embriões bovinos, embora a TC tenha sido afetada.

Effect of oils sources on blood lipid parameters of commercial laying hens

The experiment was carried out to verify if total cholesterol, HDL-cholesterol and triacylglicerol plasma levels are affected when laying hens are fed rations containing different dietary oil sources. One hundred sixty 50 week-old hens, assigned to four treatments with five replicates using 8 hens per replicate were used. The experimental period was of 84 days divided in 3 cycles of 28 days each. In the last day of each cycle, blood samples of 2 hens per replicate were randomly choose and blood samples were collected. on the other hand, blood was also collected at 7 am, 11 am and 3 pm aiming to study the daily changes of these lipids. Blood lipid parameters were not affected by different dietary oil sources (p > 0.05); however, HDL-cholesterol did change during the day, giving evidence that this lipid is indeed involved in the egg yolk formation.

Effect of supplementation of diets for quails with vitamins A, D and E on performance of the birds and quality and enrichment of eggs

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

O processo aterosclerótico em artérias de coelhos submetidos a dieta suplementada com gema de ovo: modelo experimental de baixo custo

OBJETIVO: Verificar a aterogenicidade do modelo de hipercolesterolemia por suplementação alimentar com gema de ovo em coelhos e seu uso como modelo de aterosclerose experimental de baixo custo. MATERIAL E MÉTODO: Foram utilizados 14 coelhos divididos em dois grupos de sete animais: grupo controle (G1), que recebeu ração comercial ad libitum, e grupo tratado (G2), que foi alimentado com dieta suplementada com gema de ovo. Ambos os grupos foram alimentados por 90 dias. Foram realizadas dosagens do perfil lipídico dos animais nos momentos 0, 30, 60 e 90 dias. Ao término do período experimental, os animais foram submetidos a eutanásia e retirada da aorta e de seus ramos diretos para realização de estudo anatomopatológico. RESULTADOS Apenas no grupo G2 houve aumento significativo nos níveis de colesterol total e frações. Ao exame macroscópico, foram observadas estrias gordurosas no arco aórtico e aorta abdominal e, à microscopia, acúmulos lipídicos discretos na íntima da aorta abdominal, renal, carótida, transição toracoabdominal e femoral. Portanto, a dieta com gema de ovo provocou aterosclerose leve no animal de experimentação e alterações equivalentes àquelas provocadas pelo colesterol purificado comercial quando fornecido em baixa dosagem. Assim sendo, a gema de ovo pode ser utilizada como fonte de colesterol alimentar de baixo custo em modelos de aterosclerose experimental.

Rastreabilidade da farinha de carne e ossos bovinos em ovos de poedeiras alimentadas com ingredientes alternativos

O objetivo deste trabalho foi detectar traços de farinha de carne e ossos bovinos, em ovos de poedeiras alimentadas com dietas comerciais com inclusão de ingredientes vegetais alternativos e leveduras. A detecção foi feita pela técnica dos isótopos estáveis do carbono e do nitrogênio. Foram utilizadas 384 poedeiras, distribuídas aleatoriamente em oito tratamentos. Os tratamentos consistiram de uma dieta-controle - à base de milho e farelo de soja - e sete dietas com inclusão de farinha de carne e ossos bovinos, acrescidas ou não de outros ingredientes (farelo de trigo, quirera de arroz, farelo de algodão, glúten de milho, levedura de cana e levedura de cerveja). No 35º dia, foram tomados aleatoriamente 24 ovos por tratamento: 12 para análise de ovo e 12 para análise de gema e albúmen, em separado. Após análise isotópica de carbono e nitrogênio, os resultados foram submetidos à análise multivariada de variância. As médias dos pares isotópicos dos ovos, gema e albúmen, em todos os tratamentos, diferiram daquelas do tratamento-controle. A técnica dos isótopos estáveis permite detectar, nos ovos, gema e albúmen, a farinha de carne e ossos bovinos utilizada na dieta de poedeiras, mesmo com a inclusão de outros ingredientes vegetais e leveduras.

Rastreabilidade da farinha de carne e ossos bovinos em ovos de poedeiras comerciais pela técnica dos isótopos estáveis do carbono e nitrogênio

Objetivou-se com este estudo rastrear a inclusão de farinha de carne e ossos bovinos em dietas para poedeiras comerciais, por meio da análise dos ovos e de suas frações (gema e albúmen), pela técnica dos isótopos estáveis do carbono e nitrogênio e avaliar o índice analítico mínimo detectável. Foram utilizadas 240 galinhas poedeiras da linhagem Shaver White de 73 semanas de idade, distribuídas em delineamento inteiramente casualizado, com cinco tratamentos e seis repetições. Foram avaliados cinco níveis de inclusão (0; 1,5; 3,0; 4,5 e 6,0%) de farinha de carne e ossos bovinos em uma dieta à base de milho e farelo de soja. No 35º dia, foram tomados aleatoriamente 24 ovos por tratamento: 12 serviram para amostragem de gema e albúmen e os outros 12 para amostragem do ovo (gema + albúmen). Os resultados isotópicos foram submetidos à análise multivariada de variância e, a partir das matrizes de erro, com 95% de confiança, foram determinadas elipses para identificar as diferenças entre os resultados obtidos com o fornecimento das dietas experimentais e a dieta controle, sem farinha de carne e ossos bovinos. No ovo e na gema, a partir do par isotópico da dieta com 3,0% de farinha de carne e ossos, houve diferenciação do par do tratamento controle, enquanto, no albúmen, a diferenciação ocorreu a partir do nível de 1,5% de farinha de carne e ossos bovinos na dieta. Pela técnica dos isótopos estáveis, é possível rastrear o uso de farinha de carne e ossos bovinos na alimentação de poedeiras; no albúmen, o nível mínimo de inclusão detectável é de 1,5% e, no ovo e na gema, 3,0%.

Efeito dos Níveis de Cantaxantina na Dieta Sobre o Desempenho e Qualidade dos Ovos de Poedeiras Comerciais

O experimento teve como objetivo estudar os efeitos de níveis de cantaxantina sobre o desempenho e a coloração das gemas dos ovos de galinhas poedeiras. Foram utilizadas 384 galinhas da linhagem Hisex Brown, em um delineamento em blocos ao acaso, contendo seis tratamentos (0, 12, 24, 36, 48 e 60 ppm de cantaxantina), com oito repetições de oito aves por parcela. O período experimental foi de 56 dias. A coleta de ovos foi realizada diariamente e a análise de coloração dos ovos foi efetuada com o abanico colorimétrico da Roche. Durante os 14 dias do período inicial do experimento, a melhor coloração das gemas foi obtida com a adição de 60 ppm de cantaxantina, atingindo-se a cor plateau de 14,3 do leque colorimétrico Roche aos 5,43 dias de inclusão do pigmentante. Considerando-se o período experimental total, os níveis de cantaxantina utilizados melhoraram de forma quadrática a coloração das gemas, sem influenciar os parâmetros produtivos e demais características de qualidade dos ovos de poedeiras comerciais.

Morphological and functional characteristics of chilled semen obtained from domestic feline epididymides (Felis catus)

The objective of this study was to determine morphological and functional characteristics of semen retrieved from the feline epididymis before and after cooling. Sixteen adult male cats were orchiectomized. The distal portion of the epididymis and proximal part of the deferent ducts were dissected and squeezed to obtain their content. After centrifugation, the supernatant was removed, sperm were resuspended in a 0.9 mL Tris-fructose-citric acid extender containing 20% egg yolk, aliquoted into three 0.3 mL samples, placed in a refrigerator (4.8 degrees C) and cooled (0.5 degrees C/min). Semen evaluations were performed on four occassions: immediately after epididymal sperm retrieval (TO), and at 24 h (T-1), 48 h (T-2) and 72 h (T-3) after cooling. on each occasion, progressive motility, vigor and sperm morphology were determined. Mean motility and vigor decreased (P < 0.05) between each successive examination. Although the majority of sperm cell damage occurred within the first 24 h, there was a decrease (P < 0.05) in mean percentage of morphologically normal sperm between To and each evaluated time (T-1, T-2, T-3) after cooling, due to an increase in coiled and bent sperm tails. Further studies are needed to evaluate the effects of cooling on the fertilizing capacity of cat epididymal spermatozoa in assisted reproduction programs. (c) 2006 Elsevier B.V. All rights reserved.

Evaluation of fertilizing potential of frozen-thawed dog spermatozoa diluted in ACP-106 (R) using an in vitro sperm-oocyte interaction assay

The aim of present study was to evaluate frozen canine semen with ACP-106 (R) (Powder Coconut Water) using an in vitro sperm-oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106 (R) containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106 (R) containing 20% egg yolk and 12% glycerol. Samples were thawed at 38 degrees C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 +/- 14.8% and it was significant higher than the total motility estimated by CASA (23.0 +/- 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 +/- 3.1 % and 94.3 +/- 3.1 %, respectively, post-thaw percentage of intact plasma membrane was only 35.1 +/- 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106 (R) was efficient for maintain the in vitro fertility potential of dog spermatozoa.

Prognostic value of canine frozen-thawed semen parameters on in vitro sperm-oocyte interactions

Combining the data from conventional semen analysis with oocyte penetration assays should improve the assessment of the fertilizing ability of a semen sample. Thus, the objective of the present study was to evaluate the prognostic value of various semen parameters on the in vitro interactions between frozen-thawed canine sperm and homologous oocytes. Ten ejaculates from five stud dogs (two ejaculates/dog) were collected by digital manipulation. Semen samples were evaluated, extended in Tris-egg yolk-glycerol, frozen and stored in liquid nitrogen, and thawed several weeks later. Samples were evaluated for motility and sperm populations by computer-aided semen analysis (CASA), plasma membrane integrity (carboxy-fluorescein diacetate and propidium iodide), and sperm morphology (Bengal Rose). Thawed spermatozoa were also incubated with homologous oocytes for 18 h in an atmosphere of 5% CO2 and 95% air at 38 degrees C and sperm-oocyte interactions were evaluated. Simple linear regression models were calculated, with sperm parameters as independent variables and sperm-oocyte interactions as the dependent variable. There were significant associations between: percentage of oocytes bound to spermatozoa and beat cross frequency (BCF; R-2 = 63%); percentage of oocytes that interacted with spermatozoa and BCF (R-2 = 73%); and number of penetrated spermatozoa and velocity average pathway (VAP; R-2 = 64%) and velocity straight line (VSL; R-2 = 64%). Although plasma membrane integrity and sperm morphology had little prognostic value for in vitro interactions between canine frozen-thawed sperm and homologous oocytes, some motility patterns (evaluated by CASA) were predictive of in vitro sperm-oocyte interactions. (c) 2005 Elsevier B.V. All rights reserved.

Impact of 24-h Cooling Prior to Freezing on the Survival of Domestic Cat (Felis catus) Epididymal Sperm

The aim of this study was to investigate the impact of a 24-h cooling period prior to freezing on domestic cat epididymal sperm viability. Fifteen tomcats were submitted to routine orchiectomy and sperm samples were retrieved from both epididymides in a Tris-glucose-20% egg yolk extender. For each tomcat, the diluted sperm was split into two equal volumes and cooled to 5 degrees C at a rate of 0.5 degrees C/min; one sample for 60 min (control) and the other for 24 h (cooled). After the cooling period, samples from both groups were frozen using an identical freezing protocol. Sperm samples were evaluated in three different periods: immediately after harvesting, after cooling at 5 degrees C for 24 h (cooled group) and after freezing thawing of control and cooled groups. Evaluations consisted of sperm motility and progressive status, sperm morphology and plasma membrane integrity (PMI) using two fluorescent probes. After cooling for 24 h, a decrease (p < 0.05) in sperm motility, progressive status and PMI was observed when compared to sperm samples immediately after collection. Comparing the results obtained after thawing, no difference (p < 0.05) was found regarding sperm motility, progressive status, PMI and sperm morphology between control and cooled groups. The results from the present study show that cooling cat epididymal spermatozoa at 5 degrees C for 24 h prior to freezing does not lead to major damage of spermatozoa impairing the freeze-thaw process.

Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants

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

Effect of refrigeration systems upon frozen bull sperm viability assessed by computer-assisted sperm analysis and fluorescent probes

Sperm cryopreservation success depends upon the maintenance of spermatozoa fertility potential. Sperm cells must preserve both integrity and functionality of several cell structures. The stabilization phase must allow the exit of water from the sperm cells via osmosis. This study aimed to compare the effect of refrigeration in the commercial refrigerator (CR) and the transport/refrigeration box (TRB) upon the viability of frozen bull sperm diluted in three different extenders (A, B and C). Ten Nellore bulls, Bos taurus indicus maintained in Artificial Insemination Center were used and the spermatozoa samples was assessed for Plasma Membrane Integrity and CASA evaluation. The stabilization phase (5 degrees C/4 hours) was performed in the CR as well as in the TRB, and then samples were exposed to nitrogen vapor during 20 minutes and then plunged into nitrogen. The statistical analysis was done using the variance analysis and the significance level was set at 5%. In the CR the post-thawing parameters for PM and ALH were higher (p < 0.05) in the extender A (glicine egg-yolk) and extender B (glicine egg-free) when compared with extender C (TRIS egg-yolk). As for BCF, STR and LIN, the parameters were higher (p < 0.05) in extender B than in C. Samples that were stabilized in the TRB presented higher post-thawing parameters (p < 0.05) for PM and LIN in extender A and extender B when compared with C. BCF and STR parameters were higher (p < 0.05) in extemder B when compared with C. Extender B samples had higher (p < 0.05) PMI when stabilized in CR. The findings in this experiment enable us to say that both CR and TRB were effective in keeping the viability of post-thawing bull semen.

Vitellogenin content in fat body and ovary homogenates of workers and queens of Melipona quadrifasciata anthidioides during vitellogenesis

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