Trehalose and glycerol have a dose-dependent synergistic effect on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender

P. 275-282The objective of this study was to examine the interaction of different concentrations of trehalose [0 (T0), 50 (T50) or 100 (T100) mM] and glycerol [5% (G5) or 7% (G7)] on post-thawed quality of ram semen, cryopreserved in a soybean lecithin (SL)-based extender. Twenty-eight ejaculates were collected from four rams and diluted with six trehalose/glycerol combinations: T0G5, T50G5, T100G5, T0G7, T50G7, and T100G7. Sperm motility (CASA), membrane integrity (eosin/nigrosin) and functionality (HOST), abnormal forms, capacitation status (CTC), mitochondrial activity (rhodamine 123), apoptotic features (Annexin V/propidium iodide) and lipoperoxidation (malondialdehyde production) were evaluated after thawing. Extender T100G5 yielded the highest results for total and progressive motility, sperm velocity, normal morphology, functional membranes, active mitochondria and membrane integrity, with P 0.05). The combinations T0G5, T0G7 and T100G7 yielded the lowest post-thaw quality. We could not detect significant changes in other kinematic parameters, capacitation status or lipoperoxidation. We conclude that, in our SL-based extender, a combination of 100 mM trehalose and 5% glycerol was the most adequate combination to achieving post-thawing quality in our soybean lecithin-based extender, and our results support that a synergistic effect among trehalose and glycerol exists. We suggest that other combinations could improve these results.S

The effect of acidifier supplementation on egg production performance and intestinal histology of Japanese quail (Coturnix japonica)

Abstract Background Acidifier are substances with antibacterial, antifungal, antimicrobial, performance and health benefits that are frequently employed in feed acidification, especially in poultry diet. Meanwhile, the most important factor for acidifier efficiency is the proportion of different acids in the final product. Objectives This study aimed to investigate the effect of dietary supplementation of a commercial acidifier on egg production and histology of the small intestine in laying Japanese quail. Methods One‐hundred and sixty female quails at 15 weeks of age were divided into four groups and fed basal diet supplemented with different levels of acidifier (0, 1, 2 and 3 gr acidifier/kg of basal diet) for 8 weeks. Egg production, egg quality attributes and body weight (BW) were measured every 2 weeks. Histology of the small intestine and bacterial population of cecum as well as pH of crop, duodenum, jejunum, ileum and cecum contents were also investigated at the end of the experiment. Results Feed conversion ratio (FCR), yolk height, shell thickness, pH of the duodenum, jejunum, ileum, cecum; duodenum, villus width (VW), villus height (VH), crypt depth (CD); jejunum VH, VW and ileum VH to CD ratio (VCR) were linearly improved by the increasing levels of acidifier supplementation (P  0.05). Conclusions In conclusion, as calculated, the supplementation of 1 and 2.6 g acidifier per kg of diet was associated with beneficial effects on egg production and quality, gastrointestinal tract pH and histology of the small intestine in laying quails

Oocyte quality determines bovine embryo development after fertilisation with hydrogen peroxide-stressed spermatozoa

Exposure of gametes to specific stressors at sublethal levels can enhance the gametes' subsequent performance in processes such as cryopreservation. In the present study, bull spermatozoa were subjected to H2O2 for 4 h at 100-, 200- and 500-mu M levels; computer-assisted sperm analysis (CASA) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay were used for evaluation of subsequent sperm motility and DNA integrity, respectively. Exposure of spermatozoa to H2O2 did not affect sperm motility but DNA integrity was negatively affected by 500 mu M H2O2 compared with mock-exposed spermatozoa, whereas both motility and DNA integrity were affected compared with untreated spermatozoa. Nevertheless, insemination of oocytes with spermatozoa exposed to 200 mu M H2O2 increased fertilisation, cleavage and blastocyst rates (P < 0.05). Furthermore, the higher blastocyst yield after fertilisation of oocytes with spermatozoa exposed to 200 mu M H2O2 was related to oocyte diameter, with large-medium oocytes yielding higher blastocyst rates, while small-diameter oocytes consistently failed to develop into blastocysts. In conclusion, the results indicate that exposure of spermatozoa to 200 mu M H2O2 before sperm-oocyte interaction may enhance in vitro embryo production in cattle. However, this increased embryo production is largely dependent on the intrinsic quality of the oocytes

Cervical dilation and improvement of reproductive performance in fat-tailed ewes via cervical dilator treatments

Objective: To determine the effect of different cervical dilators on cervical dilation and reproductive performance of fat-tailed ewes.Methods: In experiment 1 140 ewes were divided into seven groups with seven different treatments as following: 10 mL normal saline (control group), 100 IU oxytocin (OT group), 100 μg estradiol and 100 IU oxytocin (E2+OT group), 5 mL relaxin (R group), 2 mL sensiblex (SEN group), 200 μg misoprostol (MIS group) or 200 μg dinoprostone (DIN group). In experiment 2, artificial insemination was applied for evaluation of reproductive performance in experimental groups.Results: In experiment 1, the highest cervical dilation was observed in OT (90%) and E2+OT (100%) groups (P<0.05), while no significant differences was found among DIN, MIS, SEN and R groups (80%, 75%, 70% and 65%, respectively). In addition, the lowest cervical dilation was observed in control group. Experiment 2 found no significant differences among control, OT and E2+OT groups. The highest pregnancy rate, parturition rate and lambing rate were observed in OT groups (60%, 60% and 70%, respectively) and E2+OT groups (65%, 60% and 70%, respectively) compared to SEN, R, MIS and DIN groups (P<0.05).Conclusions: Oxytocin treatment alone or with estradiol could be used as a suitable dilator for improving reproductive efficiency during artificial insemination in fat-tailed ewes

Additional file 1 of Gene expression profile of placentomes and clinical parameters in the cows with retained placenta

Additional file 1: Table S1. Primer sequences designed for qPCRexperiments. Table S2. Information about reads and alignments. Table S3. DEGs list. Tables S4 and S5 and Fig. S1: GSEA analysis results

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

To associate glucose-6-phosphate dehydrogenase (G6PDH) activity in goat oocytes with intracellular glutathione (GSH) content, meiotic competence, developmental potential, and relative abundance of Bax and Bcl-2 genes transcripts.Purpose To associate the glucose-6-phosphate dehydrogenase (G6PDH) activity inimmature goat oocyte with intracellular glutathione (GSH) content, developmentalcompetence after parthenogenetic activation and relative abundance of Bax and Bcl-2genes transcripts using .Methods Goat oocytes were exposed to brilliant cresyl blue (BCB) staining andcategorized into BCB+ (blue cytoplasm) and BCB&minus; (colorless cytoplasm) groups. Agroup of oocytes were not exposed to BCB stain and was considered as a control.After maturation in vitro, a group of oocytes were used for determination of nuclearstatus and intracellular GSH content and another group were subjected toparthenogenetic activation followed by in vitro embryo culture.Results We found that BCB+ oocytes yielded a significantly higher maturation rate thanthe BCB&minus; and control oocytes. Moreover, BCB+ oocytes showed increased inintracellular level of GSH than BCB&minus; and control oocytes. Furthermore, BCB+ oocytesproduced more blastocysts than BCB&minus; and control oocytes. The expression of antiapoptoticgene Bcl-2 and pro-apoptotic gene Bax in mature oocyte, their surroundingcumulus cells and blastocyst was affected by BCB staining test.Conclusions These results strongly suggest that BCB+ oocytes have a higher meioticand cytoplasmic maturity (as determined by intracellular GSH content), and that BCBstaining can be used to select developmentally competent oocytes for in vitro embryoproduction.</p

Effect of selenium on growth performance and blood parameters of Holstein suckling calves

This experiment was conducted to evaluate and compare the effects of inorganic and organic selenium sources with inorganic and organic carriers on growth performance, starter feed intake, blood parameters, and the concentration of glutathione peroxidase in the blood of suckling Holstein calves. To this objective, 40 suckling Holstein calves (38.47±2.52 kg average birth weight) at 7 days of age were selected and randomly divided into four experimental groups (10 replicates). The experimental groups included control (without selenium supplement), inorganic selenium (supplemented with sodium selenate), organic selenium with inorganic carriers and, organic selenium with organic carriers. The results show that the supplementation of selenium significantly increased the glutathione peroxidase enzyme concentration (P<0.01) and can significantly reduce the concentration of plasma cholesterol (P<0.01). However, there was no significant effect of selenium supplementation on serum glucose, plasma total protein, triglyceride, and urea nitrogen concentrations. Also, none of the treatments had a significant effect on growth performance and starter feed intake

Influence of seasonal differences on semen quality and subsequent embryo development of Belgian Blue bulls

Belgian Blue bulls are more susceptible to high temperature and humidity index (THI) than most other cattle breeds. Here, we investigated whether high ambient temperature during summer affected semen quality and subsequent embryo development in Belgian Blue cattle. For this purpose, semen samples were collected from six healthy mature Belgian Blue bulls in March (Low THI group; THI between 30.6 and 56.4) and August 2016 (High THI group; maximum THI of 83.7 during meiotic and spermiogenic stages of spermatogenesis; 14-28 days prior to semen collection) respectively. Motility, morphology, acrosome integrity, chromatin condensation, viability, and reactive oxygen species production were assessed for frozen-thawed semen. Moreover, the efficiency of blastocyst production from the frozen-thawed semen samples of the two groups was determined in vitro. Blastocyst quality was determined by assessing inner cell mass ratio and apoptotic cell ratio. Fresh ejaculates showed a higher sperm concentration in low THI when compared to the high THI group (P 0.05). In frozen-thawed semen, total and progressive motility, viability, and straight-line velocity were lower in high THI compared to the low THI group (P < 0.05), while H2O2 concentration, aberrant chromatin condensation, and abnormal spermatozoa were higher in the high THI group (P < 0.05). Blastocyst rates were significantly higher when low THI samples were used (P < 0.05). Moreover, the total cell number and trophectoderm cells were significantly higher (P < 0.05) in blastocysts derived from low THI samples, whereas the apoptotic cell ratio was significantly higher (P < 0.01) in blastocysts derived from high THI spermatozoa. In summary, our data show that elevated ambient temperature and humidity during summer can decrease the quality of frozen-thawed spermatozoa in Belgian Blue bulls and also affect subsequent embryo development. Published by Elsevier Inc

Association of glucose-6-phosphate dehydrogenase activity with oocyte cytoplasmic lipid content, developmental competence, and expression of candidate genes in a sheep model

To evaluate associations of glucose-6-phosphate dehydrogenase (G6PDH) activity in sheep oocytes with cytoplasmic lipid content, maturational competence, developmental competence to the blastocyst stage, and gene expression of certain molecular markers.Purpose To evaluate associations of glucose-6-phosphate dehydrogenase (G6PDH)activity in sheep oocytes with cytoplasmic lipid content, maturational competence,developmental competence to the blastocyst stage, and gene expression of certainmolecular markers.Methods Before brilliant cresyl blue (BCB) staining test, oocytes were classified ashigh, middle, and low cytoplasmic lipid content (HCLC, MCLC, and LCLC) and after thetest as having low or high G6PDH-activity (BCB+ and BCB-, respectively). Aftermaturation in vitro, a group of oocytes were subjected to IVF followed by in vitroembryo culture and another group was used for evaluation of expression of candidategenes.Results The cleavage and blastosyst rates were lowest (P &lt; 0.05) in LCLC group,intermediate (P &lt; 0.05) in MCLC group and highest (P &lt; 0.05) in HCLC group. More(P &lt; 0.05) oocytes in HCLC group were BCB+, and higher (P &lt; 0.05) maturation,cleavage, and blastocyst rates were seen for BCB+ oocytes than the BCB- oocytes.Our gene expression data indicated that mRNA transcript abundance of ITGB2, pZP3,BMP15, and GDF9 genes were similar between BCB oocytes groups. However, theexpression of ATP1A1 was higher (P &lt; 0.05) for BCB+ oocytes compared to BCBoocytes.In addition, BAX transcript abundance was similar (P &gt; 0.05) among BCB+,BCB-, and control groups, before and after maturation in vitro.Conclusion Activity of G6PDH in sheep oocytes is highly associated with lipid content,and compared with the morphological parameters, might be a more precise andobjective predictor for subsequent developmental competence in vitro.</p