Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Carbon dynamics in small tropical catchments under preserved forest and cacao agroforestry systems

Agroforestry Systems - Inland waters such as streams that receive carbon from terrestrial landscapes usually have a net heterotrophic metabolism and emit significant amounts of CO2 to the...     

Multivariate analysis for characteristics of heat tolerance in horses in Brazil

The environment in which the horse is reared affects its ability to maintain thermal balance which is in turn related to thermal characteristics and regulatory physiological mechanisms. In this study a multivariate analysis of physiological traits in relation to heat tolerance in horses was carried out in the Federal District, Brazil. The aim was to test the ability of these analyses to separate groups of animals and determine which physiological traits are most important in the adaptation to heat stress. Forty adult horses (4 to 13 years) were used, ten from each of four different genetic groups (English thoroughbred, Brazilian showjumper, crossbred and Breton). The traits examined included heart and breathing rate, rectal temperature as well as blood parameters. The data underwent multivariate statistical analysis including cluster, discriminate and canonical using Statistical Analysis System - SAS ® procedures CLUSTER, STEPDISC, CANCORR and DISCRIM. The tree diagram showed clear distances between groups studied and canonical analysis was able to separate individuals in groups. The discriminate analysis identified the variables which were most important in separating these groups. The multivariate analysis was able to separate the animals into groups with RR, HR and RT being important in this separation.     

Fatty acid profile of hair lambs and their crossbreds slaughtered at different weights

This study aims to investigate the effect of breed and slaughter weight on fatty the acid profile of the Longissimus dorsi muscle in lambs. Twenty-four Santa Inês (SI), 24.5 Ile de France × 0.5 Santa Inês (Ile × SI) and 12.5 Texel × 0.5 Santa Inês (Te × SI), slaughtered at different weights (30, 35, 40, and 45 kg), were evaluated. The animals were reared with creep feeding to weaning and were feedlot finished, receiving a diet composed of 30% hay and 70% concentrate. The analyses of fatty acids were carried out on the Longissimus muscle of the 13th rib. The experiment was in a three by four factorial design. The total mean saturated fatty acid level was 44.88%, with 43.30% monounsaturated fats and 1.72% polyunsaturated. The major fatty acids found included oleic (43%), palmitic (22%), and stearic (18%). The concentration of desirable fatty acids varied from 61.56% to 66.78%, with Te × SI (66.78%) having the highest levels (P < 0.05). The slaughter weight affected (P < 0.05) the saturated and unsaturated (both mono and poly) fatty acid profiles. The 35-kg slaughter weight showed the most desirable fatty acid profile. The saturated myristic and stearic fatty acids decreased with an increase in the slaughter weight, but oleic acid increased. In the conditions of the present study, the meat from Santa Ines and Texel × Santa Ines lambs had fatty acid profiles more beneficial for human health due to the higher levels of unsaturated fatty acids.     

Effects of concanavalin A on the progesterone production by bovine steroidogenic luteal cells in vitro

The aim of this study was to evaluate the effects of concanavalin A (CONA) on the progesterone (P4) production by bovine steroidogenic luteal cells (LCs) in vitro. Luteal cells were collected during the mid‐luteal stage (at 10–12 days following ovulation) and processed in the laboratory. Luteal cells were grown for 7 days in a humid atmosphere with 5% CO₂, with or without 10% foetal bovine serum, and were subjected to the following treatments: control: no treatment; CONA (10 μg/ml); LH (100 μg/ml); CONA + LH; LH (100 μg/ml) + prostaglandin F2α (PGF2α) (10 ng/ml); CONA + LH + PGF2α. Samples of the culture media were collected on days 1 (D1) and 7 (D7) for P4 quantification. The cells were counted on D7 of culture. Differences between treatments were considered statistically significant at p < .05. Culture in the presence of CONA decreased the P4‐secreting capacity of LCs on D7 of culture, particularly in the absence of serum. The cell numbers did not change between treatments.     

Use of heat tolerance traits in discriminating between groups of sheep in central Brazil

The animal and its environment make up an integrated system, where each acts on the other. Tropical regions are characterized by high levels of solar radiation and environmental temperature which may adversely affect animal production. This study carries out a multivariate analysis of physical and physiological traits in sheep in the Federal District of Brazil to test the ability to separate groups of animals and determine which traits are most important in the adaptation of animal to heat stress. The variables studied included coat thickness, number and length of hairs, pigmentation of the skin and coat, number of sweat glands as well as heart and respiratory rates, rectal and skin temperatures, sweating rate, and blood parameters. Five groups of ten animals were used depending on breed (Bergamasca, crossbred, or Santa Inês) or coat color (Santa Inês—brown, white, and black). The data underwent multivariate statistical analyses including cluster, discriminate, and canonical, using Statistical Analysis System—SAS®. The tree diagram showed clear distances between groups studied and canonical analysis was able to separate individuals in groups, especially Bergamasca and white Santa Inês. The canonical correlation redundancy analysis showed that coat reflectance as well as hair length and number of hairs per unit area were the most useful in explaining changes in physiological traits. Skin and coat traits such as hair length, coat reflectance, percentage of epithelial area occupied by sweat glands, skin reflectance and thickness, as well as heart and breathing rates were the most important in separating these groups.

     

Physical,chemical and sensorial parameters for lambs of different groups,slaughtered at different weights

The object of this experiment was to study physicochemical and sensorial traits in the 11th and 13th ribs of 24 Santa Ines (SI), 24 1/2 Ile de France?×?1/2 Santa Inês (ILE?×?SI) and 8 1/2 Texel?×?1/2 Santa Inês (TE?×?SI), slaughtered at different weights (30, 35, 40 and 45 kg, respectively). Subjective measurements (marbling, colour and texture) were carried out on the Longissimus dorsi, as well as initial pH (0 h) and final pH (24 h) after slaughter. The experiment was in a 3?×?4 factorial design and analysed using general linear model and correlation procedures in SAS®. Breed group did not influence colour (3.32), shear force (3.57 kg-force (kgf)) or loss in cooking (24.05%) of the L. dorsi, but slaughter weight affected these, with stronger colour, tougher meat and greater loss in cooking as slaughter weight increased. For sensorial analysis, the L. dorsi were cut, identified and evaluated by 30 untrained judges using a linear scale. Significant differences were detected in preference (6.61 points), tenderness (6.32 points), succulence (6.33 points) and flavour (7.08 points) of the meat from different breed groups and slaughter weights. For preference, all meats were well accepted although the crossbred animals slaughtered at 45 kg were less acceptable.     

Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification

This study aimed to produce in vitro bovine embryos by the addition of two drugs, which is responsible for oocyte meiosis inhibition: roscovitine (ROS) and butyrolactone I (BL‐I). Oocytes were recovered from slaughtered cows and matured in a commercial medium and maintained in a 5% CO₂ atmosphere. Oocytes were maintained for 6 h in an in vitro maturation (IVM) medium containing ROS (12.5 μm ), BL‐I (50 μm ) and association of drugs (ROS 6.25 μm and BL‐I 25 μm ). Oocytes were cultured for 18 h in an agent‐free medium for the resumption of meiosis. After 24 h of maturation, oocytes were inseminated in the commercial in vitro fertilization (IVF) medium. Presumptive zygotes were cultured in SOFaa medium in a 5% CO₂ atmosphere. On day 3, rate of cleavage was evaluated and on days 6 and 7, rate of blastocyst formation. BL‐I and its association with the ROS increased the rates of cleavage and blastocyst formation (p < 0.05). The ROS alone was inefficient, impairing embryonic development, with low rates of blastocyst formation when compared to the control group and other treatments (p < 0.05). The embryos from BL‐I and ROS+BL‐I groups presented higher number of cells and lower rates of cellular apoptosis compared to other groups, either for the fresh or for post‐thawing embryos. Embryos from ROS+BL‐I group showed to be more resistant to the vitrification process, presenting a higher rate of embryonic re‐expansion (p < 0.05). In conclusion, block of meiosis using BL‐I or its association with ROS increased the rate of blastocyst formation, and the association of ROS+BL‐I resulted in a better resistance to the embryo cryopreservation process.     

Use of a Piezo Drill for Intracytoplasmic Sperm Injection into Cattle Oocytes Activated with Ionomycin Associated with Roscovitine

Intracytoplasmic sperm injection (ICSI) consists of the introduction, by micromanipulation, of a single sperm into the cytoplasm of a mature egg. This technique is particularly advantageous when only a few sperm are available for fertilization, representing an important tool in preserving genetic material, especially from poorly fertile males. The results from ICSI in cattle are very often unsatisfactory and difficult to reproduce. Thus, the goal of this study was to evaluate the effect of the use of a Piezo drill (PD) and oocyte activation with ionomycin + roscovitine (I + R) during ICSI in cattle oocytes. After in vitro maturation (24 h), cumulus complex oocytes were divided into four groups: G1 – the ICSI was performed without the use of a PD and the oocyte was activated with I + R; G2 – the ICSI was performed with the use of the PD and activation with I + R; G3 – the ICSI was performed with the use of the PD, but without activation and G4 – parthenogenetic control, treated with I + R, but without sperm injection. The presumptive zygotes were cultured for 7 days and evaluated on day 3 for cleavage rate and on day 7 for blastocyst formation. Embryo production by standard in vitro fertilization in the laboratory was 78% for cleavage (117/150) and 35% for blastocyst formation (41/150). The cleavage rates obtained in G1, G2 and G4 were similar (66.7%, 71.6% and 66.3%, respectively), demonstrating the beneficial effect of oocyte activation. However, in G3, despite the presence of the sperm and the electric stimulation of a PD, the cleavage rates were significantly lower (17.5%) compared with the groups that used chemical activation, even in the absence of sperm (G4). Despite the beneficial effects of activation, this stimulus alone, or in the absence of the PD, was not sufficient for adequate morulae formation (13.4%, 37.9%, 0.0% and 13.5% for G1, G2, G3 and G4, respectively). Only in G2, when the PD was used followed by artificial activation, blastocysts were obtained (14.7%). These results indicate that cattle oocytes must be activated after ICSI to produce viable embryos.