Selection of X chromosome of buffaloes sperm with Percoll gradients

The aim of the present study was to evaluate the selection of X chromosome of buffaloes sperm with Percoll gradients. The stock solution of Percoll was prepared in the proportion of 1:11 (1 part of Percoll:11 parts of a solution containing KCl 1M, NaH(2)PO(4) 0.1M, NaCl 1.5M and sodium HEPES 23.8 g/ml). In order to prepare 9 different gradients were added to the stocked Percoll the A solution (glicine-yolk extender) in the following proportions: 90, 80, 72, 65, 57, 49, 34 and 25%. A sample of 0.7 ml of the fresh semen was deposited at 2 ml of Percoll 80% for the sperm wash. The precipitate was put in tube with 0.7 ml of each gradient. Then, the precipitated was washed in TES solution by centrifugation (500xg for 10 minutes), and collected again and diluted in TES solution to be freeze. The presence of the F body in the spermatozoa was observed in 58.7 +/- 5.4% of the control group and in 41.2 +/- 5.4% of the treated group (p<0.01). This result showed an increment of 17.55 of male sperm in the Percoll's group. The reduction of the centrifugation force did not improve the percentage of X sperm.

Ritmo Circadiano de Triiodotironina (T3) e Tiroxina (T4) Plasmática em Ovelhas Ideal durante o Anestro Estacional

The objective was to determine the plasma levels of the triiodothyronine (T-3) and thyroxine (T-4), in six Ideal ewe sheeps during seasonal anoestrus, by radioimmunoassay. The blood samples were collected from the jugular vein catheter over a period of 24 hours, at 2-h intervals (12, 14, 16, IX, 20, 22, 24, OZ, 04, 06, OX, and 10 hours). II was used analysis of variance for data adjustments, and the decomposition of the collected periods in non-orthogonal polynomial. It was observed a crescent linear effect of the T-4 plasma levels, presenting lower level of 49.60+/-11.41 and higher level of 76.99+/-10.0 ng/ml, at fourth and last blood samples collection, respectively. T-3 plasma levels showed a significant cubic response during 24 hours, with the highest value of 0.94+/-0.20 ng/ml at the end of the the study. In conclusion, thyroidal hormones levels were lower in the first blood samples collection, increasing progressively during the night, reaching the highest levels at the morning period.

Performance, incidence of metabolic disturbances and endocrine variables of food-restricted male broiler chickens

1. This experiment was carried out to evaluate the productive and physiological consequences of a slight but long term food restriction of male broiler chickens from 2 commercial strains.2. Cobb-500 and Ross chickens were submitted to a 20% food restriction from 8 to 21 d of age. Strain, food programme and their interactive effects were analysed in terms of consequences upon performance, mortality, incidence of sudden death syndrome (SDS) and ascites syndrome (AS), index of right cardiac hypertrophy and plasma concentrations of hormones related to metabolism and growth (T-3, T-4, T-3:T-4 ratio, IGF-I and GH).3. Although some catch-up growth was observed by refeeding previously restricted birds after 22 d of rearing, food restriction decreased (P less than or equal to 0.05) body weight at market age (42 d) irrespective of the strain, but improved (P less than or equal to 0.05) food conversion.4. The incidence of mortality was not high in non-restricted birds but SDS and AS caused more than 50% of deaths. Hypertrophic cardiac index was observed in chickens of both strains after 4 weeks of age and was higher in ad libitum fed birds.5. During the period of food restriction, plasma T-3 and IGF-I concentrations decreased whereas plasma T-4 and GH concentrations increased compared to those of the age-matched ad libitum fed counterparts. During the subsequent ad libitum feeding period, few differences in circulating hormone concentrations were observed, except for the higher mean CH litres in previously food-restricted chickens at 35 d of age.6. These results indicate that even a non-severe food restriction negatively affects body weight of 42-d-old male broilers but these are benefits with improved food efficiency and diminished mortality from metabolic disturbances. The hormone results suggest that the degree of food restriction applied was not severe because there was a very fast adaptive response with small and transient alterations in T-3, T-4 and GH plasma concentrations during the period of compensatory growth.

Productive consequences of fasting neonatal chicks of different genetic constitutions for growing

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

The importance of endogenous nutrition of chicks from divergent strains for growing tested by deutectomy

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

Effect of organic mineral supplementation on the egg quality of semi-heavy layers in their second cycle of lay

This study was carried out to evaluate the effects of dietary trace mineral levels and sources on egg quality parameters of second-cycle semi-heavy layers. A number of 360 72-week-old layers were submitted to forced molting. Upon return of lay (83 weeks of age), birds were distributed according to a completely randomized experimental design of six treatments with six replicates of 10 birds each. The control treatment consisted of 0.10% dietary supplementation of trace minerals from inorganic sources, which was proportionally replaced by five levels (110, 100, 90, 80, 70%) of an organic trace mineral supplement containing 30, 30, 40, 6, 0.61, and 0.3 g/kg product of Zn, Fe, Mn, Cu, I, and Se, respectively. All diets contained equal protein, energy, and amino acid levels. Every 28 days of the experimental period (112 days) four eggs per replicate were collected for egg quality evaluation. The following parameters were evaluated: specific gravity, yolk, albumen and eggshell percentages, yolk index, Haugh units, and eggshell thickness and breaking strength. One sample per replicate, consisting of the pool of the yolks of three eggs collected at the end of each experimental period, was used to assess protein and mineral (Ca, P, Cu, Fe, Mn, and Zn) contents. The results were submitted to ANOVA, and means to the test of Tukey at 5% significance level. The evaluated trace mineral levels and sources did not influence any of the studied egg quality parameters. It was concluded that reducing organic trace mineral supplementation in up to 70% relative to 100% inorganic trace mineral supplementation does not affect egg parameters and therefore, can be applied to the diet of semi-heavy layers in their second cycle of lay.

Calcium and available phosphorus levels for laying hens in second production cycle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cholesterol levels and nutritional composition of commercial layers eggs fed diets with different vegetable oils

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

Structural analysis of the embryonic development in Brycon cephalus (Gunther, 1869)

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

Níveis de proteína bruta e de aminoácidos sulfurados totais sobre o desempenho, a qualidade dos ovos e a excreção de nitrogênio de poedeiras de ovos marrons

O experimento foi conduzido com o objetivo de maximizar a produção e a qualidade dos ovos e minimizar a excreção de nitrogênio nas excretas de poedeiras no final do primeiro ciclo de produção, por meio do fornecimento de aporte adequado de proteína bruta (PB) e aminoácidos sulfurados totais (AAST) na dieta. Foram utilizadas 432 poedeiras Isa Brown, com 52 semanas de idade, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 3 x 3 (PB e AAST) e nove tratamentos (14 e 0,57; 14 e 0,64; 14 e 0,71; 15,5 e 0,57; 15,5 e 0,64; 15,5 e 0,71; 17 e 0,57; 17 e 0,64; 17 e 0,71 % de PB e AAST, respectivamente), com seis repetições de oito aves cada. A duração do experimento foi de 140 dias. Foram avaliadas as características de desempenho, qualidade dos ovos e excreção de nitrogênio nas excretas. A única característica de desempenho influenciada pelos tratamentos foi o peso dos ovos, que apresentou os maiores valores para as combinações de 15,5 e 0,71; 17 e 0,71; 15,5 e 0,64; 14 e 0,71 e 17 e 0,64% de PB e AAST, respectivamente. Não foram observadas diferenças significativas para consumo de ração, porcentagem de postura e de ovos quebrados, massa de ovos, conversão alimentar por dúzia e por massa de ovos e mortalidade. Para os parâmetros de qualidade dos ovos, foram observadas diferenças significativas apenas para as porcentagens de gema e de albúmem. A excreção de nitrogênio foi maior nas aves alimentadas com as rações contendo 17% de PB. Pode-se sugerir que a ração contendo 14% de PB e 0,57% de AAST pode ser utilizada, sem prejuízos no desempenho e na qualidade dos ovos, e ainda contribui para a redução da excreção de nitrogênio no ambiente e do custo da ração.

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.

Circadian and circannual rhythms of T3 and T4 secretions in Polwarth-Ideal rams

Plasma concentrations of triiodothyronine (T-3) and thyroxine (T-4) in five adult Polwarth-ldeal rams located at latitude 22degrees51 'S and longitude 48degrees26'W were evaluated every 2 months for 1 year (June, August, October, December, February, April). Blood collections were made at 2 h intervals for 24 h in each month, and hormone determinations were by radioimmunoassay. Means of T-3 (97.52 +/- 21.45 ng/dL) and T-4 (4.30 +/- 0.94 mug/dL) varied in peaks throughout the 24 h period with the highest concentrations occurring in the afternoon (16:30 and 14:30 h, respectively), and throughout the year where the highest levels were during months of long daylengths (October, December, February). Results suggest circadian and circannual rhythms in thyroid hormone secretion may be present in rams kept relatively close to the equator. (C) 2002 Published by Elsevier B.V. B.V.

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.

Effects of fetuin on zona pellucida hardening, fertilization and embryo development in cattle

Mammalian oocytes can undergo spontaneous meiotic maturation when they are liberated from their follicles and cultured in vitro; however, the zona pellucida (ZP) becomes resistant to chymotrypsin digestion, or hardens, when spontaneous maturation occurs in serum-free medium. Schroeder et al. [Biol. Reprod. 43 (1990) 891] described that fetuin, a component of fetal calf serum (FCS), inhibits ZP hardening during oocyte maturation. The aim of this experiment was to study the effect of the presence of cumulus cells and addition of hormones to maturation media on bovine zona hardening and embryo development in medium with and without fetuin. In Experiment 1, different concentrations of fetuin were added to the maturation medium. The time necessary for digestion of 50% of the ZP (d50) was not different when oocytes were matured in presence of 10% FCS, 1 mg/ml polyvinyl alcohol (PVA), or 4, 1 and 0.25 mg/ml of fetuin; cleavage rates were also similar. However, significantly more blastocysts (P < 0.05) were formed when FCS was used compared to PVA and 0.25 mg/ml of fetuin. In Experiment 11, we examined the influence of the presence of cumulus cells and hormones during the maturation of oocytes in media with PVA, BSA, FCS and fetuin. The d50 was significantly higher (P < 0.05) when oocytes were matured in presence of cumulus cells. The cleavage rate of cumulus-intact oocytes was similar for all groups. However, when oocytes were partially stripped before maturation, the cleavage rate was significantly higher (P < 0.05) when FCS or fetuin was used. In both stripped and non-stripped groups, significantly more blastocysts (P < 0.05) were formed when oocytes were matured with FCS compared to BSA and PVA. These results indicate that zona hardening, as described for mouse and human oocytes, does not have a large effect on bovine cumulus-intact oocytes. Apparently fetuin can be used as a substitute for FCS during bovine oocyte maturation, since it leads to similar developmental rates as FCS in intact and partially stripped oocytes. (C) 2002 Published by Elsevier B.V. B.V.