The effects of macromolecular and serum supplements and oxygen tension during bovine in vitro procedures on kinetics of oocyte maturation and embryo development

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

Postharvest Conservation of Mature-Green and Ripe 'Paluma' Guava Stored at Two Temperatures

This work aimed to establish the importance of maturation and ripeness stages and the use of refrigeration for the conservation of 'Paluma' guavas. Fruit picked at the mature-green and ripe stages were stored at ambient conditions (21 degrees C and 85% RH) and also at 10 degrees C (85% RH). The fruit were evaluated every 2 or 3 days for weight loss, appearance, decay, color, firmness, soluble solids, titratable acidity, ascorbic acid, total polyphenols extractable content and total antioxidant activity. The fruit stored at 21 degrees C had higher weight loss than those stored at 10 degrees C. Mature-green guavas at 21 degrees C remained in good quality for 6 days, but at 10 degrees C, the preservation period increased to 15 days. Ripe fruit were preserved for 4 days at 21 degrees C, which was extended with refrigeration to 6 days. Mature-green fruit at 21 degrees C had decay in 6 days; while at 10 degrees C decay happened in 18 days. The peel color of mature-green fruits, at 21 degrees C, showed increasing values of luminosity, indicating that its color became lighter (change from green to yellow) and at 10 degrees C it showed constant values until the end of storage. Pulp firmness of mature-green fruit declined during storage as a result of ripening. In ripe fruits such reduction occurred more slowly, since they were softer. The color of the pulp became intense red for mature fruits. Soluble solids were lower in ripe fruit at 21 degrees C, while in mature fruits at 10 degrees C, it increased. The titratable acidity increased in fruits stored at 10 degrees C. The fruits kept at 21 degrees C and the mature guavas kept at 10 degrees C showed no changes in ascorbic acid content. The ripe fruit at 10 degrees C maintained their ascorbic acid levels. Mature guavas, stored at 10 degrees C, had the longest shelf life and higher contents of soluble solids and titratable acidity, with no changes in total polyphenols extractable content and total antioxidant activity.

Models for genetic evaluation of Nelore cattle mature body weight

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic associations of conformation, finishing precocity and muscling visual scores with mature weight in Nelore cattle

Were estimate (co)variance and genetic associations between conformation, finishing precocity and muscling visual scores measured at weaning (SCW, SFW and SMW, respectively) and yearling (SCY. SFY and SMY, respectively) with mature weight (MW) in Nelore cows, in order to predict the possible changes that inclusion of visual scores in beef cattle selection indices would bring to female mature weight. The data set contained records of 36,757 females, born between 1993 and 2006, belonging to the Jacarezinho cattle raising farm. (Co)variance components were estimated by bivariate animal models using Bayesian inference method through Gibbs sampling, assuming a linear model for MW and a nonlinear (threshold) model for conformation, finishing precocity and muscling visual scores. The first 10,000 rounds were considered as the burn-in period and discarded. The posterior means of direct heritability distributions were: 0.16 +/- 0.02 (SCW); 0.20 +/- 0.02 (SFW); 0.19 +/- 0.02 (SMW); 0.24 +/- 0.02 (SCY); 0.31 +/- 0.02 (SFY); 0.32 +/- 0.02 (SMY) and 0.46 +/- 0.04 (MW). Estimates of genetic correlations between visual scores and MW were positive and moderate, ranging from 0.27 +/- 0.06 to 0.36 +/- 0.04. Visual scores and MW should respond favorably to direct selection. Mature weight can be used in Nelore breeding programs designed to monitor the cows' size. Selection of animals with higher conformation, finishing precocity and muscling scores, especially at yearling, should promote an increase in cows' mature weight. (c) 2010 Elsevier B.V. All rights reserved.

Produção in vitro de embriões bovinos: utilização de diferentes fontes de gonadotrofinas na maturação dos oócitos

O presente trabalho foi conduzido com o objetivo de avaliar o efeito da utilização de diferentes fontes de gonadotrofinas para maturação in vitro dos oócitos bovinos fecundados e desenvolvidos in vitro sobre as taxas de clivagem (TC) e de blastocistos (TBL). Oócitos imaturos provenientes de ovários de vacas de abatedouro foram submetidos a maturação in vitro sob diferentes condições: meio TCM 199, acrescido de 10% de soro de vaca em estro (SVE), aditivos, hepes, NaHCO3, piruvato de sódio, antibióticos (meio B-199), 20 UI/mL de PMSG e 10 UI/mL de hCG (PMSG/hCG) ou meio B-199, acrescido de 5 mig/mL de FSH e 5 mig/mL de LH (FSH/LH). Seguidos 24 h de cultura a 38,5ºC em atmosfera com 5% de CO2, os oócitos maturos foram incubados com sêmen descongelado durante 18 a 21 horas. Após esse período, os oócitos foram transferidos para placas contendo microgotas de meio Ménezo suplementado com 10% de SVE e células epiteliais do oviduto bovino em suspensão, cobertas com óleo de silicone, os quais permaneceram em cultura por mais 9 dias. Os dados foram analisados pelo teste do Qui-quadrado. A TC e a TBL, para PMSG/hCG e FSH/LH, foram 60 e 13,9% e 61,2 e 10,6%, respectivamente. Não houve diferença entre os tratamentos com relação a TC ou a TBL. Esses resultados sugerem que ambas as fontes de gonadotrofinas podem ser utilizadas para maturação in vitro dos oócitos fecundados e desenvolvidos in vitro.

Genetic analysis of visual scores and their relationships to mature female weight in Nellore breed

Dados de 79.884 animais da raça Nelore foram utilizados para estimar parâmetros genéticos e avaliar as relações entre os escores de conformação, precocidade e musculatura obtidos à desmama e ao sobreano e o peso das fêmeas à idade adulta. Utilizou-se o método da máxima verossimilhança restrita, em análise multicaracterísticas, com modelo que incluiu os efeitos genéticos aditivos direto e residual, como aleatórios, e os efeitos fixos de grupo de contemporâneos e, como covariáveis, a idade do animal à pesagem e a idade da mãe ao parto (exceto para o peso das fêmeas à idade adulta). Os grupos contemporâneos à desmama foram definidos pelas variáveis: sexo, rebanho, ano e mês de nascimento, grupo de manejo ao nascimento e à desmama. Na definição de grupo contemporâneo ao sobreano também foi incluída a variável grupo de manejo ao sobreano. Para o peso das fêmeas à idade adulta, o grupo de contemporâneos foi composto por rebanho, ano de nascimento, grupo de manejo ao sobreano, ano e estação da pesagem. Os efeitos genético materno e de ambiente permanente materno também foram incluídos no modelo para análise dos escores de conformação, precocidade e musculatura à desmama. As estimativas de herdabilidade direta obtidas foram 0,18 ± 0,02 para o escore de conformação; 0,21 ± 0,01 para o escore de precocidade; 0,22 ± 0,01 para o escore de musculatura à desmama e 0,24 ± 0,01 para o escore de conformação; 0,27 ± 0,01 para o escore de precocidade; e 0,26 ± 0,01 para o escore de musculatura ao sobreano e 0,42 ± 0,02 para o peso das fêmeas à idade adulta. As correlações genéticas estimadas entre os escores visuais medidos à desmama e ao sobreano foram positivas, variando de média a alta magnitude (0,56 ± 0,03 a 0,85 ± 0,01). Por outro lado, as correlações genéticas estimadas entre os escores visuais e o peso das fêmeas à idade adulta foram positivas e moderadas, variando de 0,21 ± 0,03 a 0,35 ± 0,03. Os resultados obtidos indicam que a seleção de animais com maiores escores visuais, principalmente ao sobreano, deve promover aumento do peso das fêmeas à idade adulta.

Genetic parameters and relationships of heifer pregnancy and age at first calving with weight gain, yearling and mature weight in Nelore cattle

The objectives of the current study were to investigate the additive genetic associations between heifer pregnancy at 16 months of age (HP16) and age at first calving (AFC) with weight gain from birth to weaning (WG), yearling weight (YW) and mature weight (MW), in order to verify the possibility of using the traits measured directly in females as selection criteria for the genetic improvement of sexual precocity in Nelore cattle. (Co)variance components were estimated by Bayesian inference using a linear animal model for AFC, WG, YW and MW and a nonlinear (threshold) animal model for HP16. The posterior means of direct heritability estimates were: 0.45 +/- 0.02; 0.10 +/- 0.01; 023 +/- 0.02; 0.36 +/- 0.01 and 0.39 +/- 0.04, for HP16, AFC, WG, YW and MW, respectively. Maternal heritability estimate for WG was 0.07 +/- 0.01. Genetic correlations estimated between HP16 and WG, YW and MW were 0.19 +/- 0.04; 0.25 +/- 0.06 and 0.14 +/- 0.05, respectively. The genetic correlations of AFC with WG, YW and MW were low to moderate and negative, with values of -0.18 +/- 0.06; -0.22 +/- 0.05 and -0.12 +/- 0.05, respectively. The high heritability estimated for HP16 suggests that this trait seem to be a better selection criterion for females sexual precocity than AFC. Long-term selection for animals that are heavier at young ages tends to improve the heifers sexual precocity evaluated by HP16 or AFC. Predicted breeding values for HP16 can be used to select bulls and it can lead to an improvement in sexual precocity. The inclusion of HP16 in a selection index will result in small or no response for females mature weight. (C) 2011 Elsevier B.V. All rights reserved.

Determination of fiber length and juvenile and mature wood zones from Hevea brasiliensis trees grown in Brazil

The aim of this study was to verify the average fiber length and the juvenile and mature wood zones from Hevea brasiliensis proveniences from a reforestation area in São Paulo, Brazil. For this purpose, five Hevea brasiliensis trees were randomly collected from a 50-year-old plantation, located in Balsamo, São Paulo, Brazil. The trees were cut and five flat sawn boards were obtained. The juvenile and mature wood zones were determined by fiber length measurement from the pith to the bark. The results showed that: (a) the juvenile wood of this species occurred approx. between 40 and 55 mm, from the pith, and from this point forward, the mature wood zone was found; (b) there was a significant difference between the average fiber length of juvenile wood (1.26 mm) and mature wood (1.51 mm).

Physical and chemical changes in juvenile and mature woods of Pinus elliottii var. elliottii by thermal modification

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

Recombinant luteinizing hormone supplementation to recombinant follicle-stimulation hormone during induced ovarian stimulation in the GnRH-agonist protocol: A meta-analysis

Purpose: to compare the efficacy of recombinant LH supplementation for controlled ovarian stimulation in recombinant FSH and GnRH-agonist protocol.Methods: Search strategies included on-line surveys of databases. The fixed effects model was used for odds ratio and effect size (weighted mean difference). Four trials fulfilled the inclusion criteria.Results: a fewer days of stimulation (p < 0.0001), a fewer total amount of r-FSH administered (p < 0.0001) and a higher serum estradiol levels on the day of hCG administration (p < 0.0001) were observed for the r-LH supplementation protocol. However, differences were not observed in number of oocyte retrieved, number of mature oocytes, clinical pregnancy per oocyte retrieval, implantation and miscarriage rates.Conclusions: more randomized controlled trials are necessary before evidence-based recommendations regarding exogenous LH supplementation in ovarian stimulation protocols with FSH and GnRH-agonist for assisted reproduction treatment can be provided.

Artificial oocyte activation using calcium ionophore in ICSI cycles with spermatozoa from different sources

The present study evaluated the effect of artificial oocyte activation (AOA) with calcium ionophore A23187 oil intracytoplasmic sperm injection (ICSI) cycles using spermatozoa from different sources. The 314 cycles evaluated were divided into three groups according to sperm origin, the ejaculated group (n = 92), the epididymal group (n = 82). and the testicular roup (n = 140). Each group was further split into experimental subgroups, depending oil whether or no AOA was performed. In additions the cycles of women younger than 36 years were evaluated separately. For each experimental group, ICSI outcomes were compared between subgroups. No significant difference was observed between subgroups for all sperm origin groups. When evaluating only the cycles of women younger than 36 years of age, AOA increased the percentage of high-quality embryos (74.5 versus 53.0%. P = 0.011) and the implantation rate (19.3 versus 10.5%, P = 0.0025) when it was used with ejaculated spermatozoa, and the percentage of high-quality embryos (64.4 versus 50.3%, P = 0.006) when epididymal spermatozoa were used. These results may suggest that both sperm maturity and oocyte quality play a role in oocyte activation. However. this study is to be continued to confirm these findings.

Artificial oocyte activation with calcium ionophore A23187 in intracytoplasmic sperm injection cycles using surgically retrieved spermatozoa

Objective: To evaluate the effect of artificial oocyte activation (AOA) on intracytoplasmic sperm injection (ICSI) cycles using surgically retrieved sperm.Design: Laboratory study.Setting: Fertility/assisted fertilization center.Patient(s): Couples undergoing surgical sperm retrieval for ICSI (n = 204).Intervention(s): Application of calcium ionophore A23187 for AOA.Main Outcome Measure(s): Cycles were divided into experimental groups according to the origin of the sperm used for injection and the type of azoospermia: [1] testicular sperm aspiration in nonobstructive-azoospermic patients (TESA-NOA group, n = 58), [2] TESA in obstructive-azoospermic patients (TESA-OA group, n = 48), [3] and percutaneous epididymal sperm aspiration in obstructive-azoospermic patients (PESA-OA, n = 98). For each experimental group, cycles where AOA was applied (subgroup: activation) were compared with cycles in which AOA was not applied (Subgroup: control). The fertilization, high-quality embryo, implantation, and pregnancy rates were compared among the subgroups.Result(s): For patients undergoing TESA, AOA did not improve ICSI outcomes for either type of azoospermia. However, for cases in which the injected sperm were retrieved from the epididymis, a statistically significantly increased rate of high-quality embryos was observed with AOA.Conclusion(s): Artificial oocyte activation may improve ICSI outcomes in azoospermic patients when epididymal, but not testicular spermatozoa, are injected. (Fertil Steril (R) 2009;92:131-6. (C)2009 by American Society for Reproductive Medicine.)

Decreased fertility in poor responder women is not related to oocyte morphological status

Introduction: In women showing impaired fertility, a decreased response to ovarian stimulation is a major problem, limiting the number of oocytes to be used for assisted reproduction techniques (ART). Despite the several definitions of poor response, it is still a matter of debate whether young poor responder patients also show a decrease in oocyte quality. The objective in this study was to investigate whether poor ovarian response to the superstimulation protocol is accompanied by impaired oocyte quality.Material and methods: This study included 313 patients younger than 35 years old, undergoing intracytoplasmic sperm injection. Patients with four or fewer MII oocytes (poor-responder group, PR, n = 57) were age-matched with normoresponder patients (NR, n = 256).Results: A higher rate of oocyte retrieval and a trend towards an increase in MII oocyte rate were observed in the NR group when compared to the PR group (71.6 +/- 1.1% and 74.1 +/- 1.0% vs. 56.3 +/- 2.9% and 66.5 +/- 3.7%; p < 0.0001 and p = 0.056, respectively). A trend toward increased implantation rates was observed in the NR group when compared to the PR group (44 and 24.5 +/- 2.0% vs. 28.8 and 16.4 +/- 3.9%; p = 0.0305 and p = 0.0651, respectively).Conclusions: Low response to ovarian stimulation is apparently not related to impaired oocyte quality. However, embryos produced from poor responder oocytes show impaired capacity to implant and to carry a pregnancy to term.