A soy lecithin nanoparticles-based extender effectively cryopreserves Holstein bull sperm.

Plant-based semen extenders, typically derived from soybean lecithin, are easier to modulate more and consistent in their composition than animal-based extenders. As large lecithin particles can, however, reduce effectiveness and solubility in bull semen extenders, sonication was used to create nano-lecithin (NL) particles of soybean lecithin. The objective was to determine the effects of lecithin type and concentration on the quality of frozen-thawed bovine sperm. We hypothesized that reducing the size of lecithin improves its interactions with the sperm and enhances the parameters that favor its motility, viability and fertility. Semen was collected from six mature Holstein bulls and ejaculates meeting minimum standards were pooled. Eight Tris-based extenders that contained 1, 2, 3, or 4 % of either conventional lecithin (L1-L4) or NL (NL1-NL4), plus two control extenders (one animal-based extender containing 20 % egg yolk [EY] and a commercial lecithin-based extender [BioXcell®]) were compared. Among soybean lecithin-based extenders, NL3 had the highest total and progressive sperm motility, and average path, straight-line and curvilinear sperm velocity, and was comparable to EY. Additionally, sperm mitochondrial activity was the highest in NL3, whereas sperm viability was highest in EY, NL3, and L4. Following in vitro fertilization of in vitro-matured bovine oocyes, NL3 had cleavage and hatching rates comparable to BioXcell®, but a lower blastocyst rate than EY. Overall, NL3 performed better than the other extenders for most end points, with efficiency comparable to EY. We, therefore, concluded that reducing lecithin particle size to a nano level improves sperm cryopreservation with optimal performance with 3 % NL.

The influence of L-proline and fulvic acid on oxidative stress and semen quality of buffalo bull semen following cryopreservation.

BACKGROUND: This study investigates the effects of cryopreservation and supplementation of Azeri water buffalo's semen with proline (Lp) and fulvic acid (FA). OBJECTIVES: Therefore, this study aimed to assess motility parameters, sperm viability, oxidative stress parameters, and DNA damage to detect the optimum concentrations of Lp and FA for buffalo semen cryopreservation. METHODS: Thirty semen samples of three buffalo bulls were diluted in Tris-egg yolk extender and divided into 12 equal groups including control (C), Lp-10, Lp-20, Lp-40, Lp-60, Lp-80 (containing 10, 20, 40, 60, 80 mM L-proline, respectively), FA-0.2, FA-0.5, FA-0.8, FA-1.1, FA-1.4 and FA-1.7 (containing 0.2%, 0.5%, 0.8%, 1.1%, 1.4% and 1.7% fulvic acid, respectively). RESULTS: The velocity parameters, TM and PM were improved by FA-1.7, FA-1.4, Lp-40 and Lp-60 groups compared to the C group but no significant difference was found regarding the amplitude of lateral head displacement and straightness compared to the control groups. The percentage of sperm viability and PMF were increased by FA-1.7, FA-1.4, FA-1.1, Lp-40 and Lp-60 groups compared to C group, while in terms of sperm DNA damage FA-1.7, FA-1.4, FA-1.1, Lp-10, Lp-20, Lp-40 and Lp-60 groups showed better results compared to C group. The results also showed that FA-1.7, FA-1.4, FA-1.1, Lp-20, Lp-40 and Lp-60 groups could improve TAC, SOD, GSH and decrease MDA levels. Also, FA-1.7, FA-1.4, Lp-20 and Lp-40 groups could improve GPx levels but just FA-1.7, and Lp-40 groups could improve CAT levels compared to C group. CONCLUSIONS: Thus, it can be concluded that L-proline and fulvic acid supplementations can improve the quality parameters of post-thawed buffalo bull semen.

Effects of dietary supplementation of tomato pomace (Solanum lycopersicum L.) and L-Arg on reproductive performance of aged male broiler breeders.

This study aimed to investigate the effects of different dietary supplementation of tomato pomace (TP) and L-arginine (L-Arg) supplementation on sperm characteristics, reproductive performance, and semen biochemical components of aged commercial male broiler breeders. Thirty Ross 308 male broiler breeders (58 wk old) were provided and assigned to 5 dietary treatment groups, including control (CON), 5% TP (TPS-5), 10% TP (TPS-10), 15% TP (TPS-15), and L-Arg supplemented (10% above the recommendation, LAS-10). The results indicated that the semen volume increased in the TPS-15 group compared to that of the LAS-10 (and CON on wk 9) throughout the study (P < 0.05). The sperm concentration significantly increased in TPS-10 and TPS-15 groups in comparison to the other experimental groups. On wk 5 and 7, the sperm viability increased in all TPS groups compared to the CON and LAS-10, while on wk 9, it only increased in the TPS-10 group in comparison to the LAS-10 group (P < 0.05). The hypo-osmotic swelling test decreased in the LAS-10 group compared to the other experimental groups on wk 5 and all TPS groups on wk 7 and 9 (P < 0.05). The sperm total motility and forward progressive motility decreased in the LAS-10 group compared to the other experimental groups (P < 0.05). In contrast, unprogressive motility and immotile sperms were increased in the LAS-10 group compared to the other experimental groups (P < 0.05). In addition, the sperm penetration and fertility rate increased in TPS-10 and TPS-15 groups in comparison to CON and LAS-10 groups (P < 0.05). However, hatchability was reduced in the LAS-10 group (P < 0.05). The semen adenosine triphosphate increased in TPS-10, TPS-15, and LAS-10 groups compared to the CON (P < 0.05). Finally, the semen TAC and superoxidase dismutase decreased in the LAS-10 group (P < 0.05), while the glutathione peroxidase increased in the TPS-15 group (P < 0.05). In conclusion, 15% dietary TPS is recommended to improve the reproductive performance of aged commercial male broiler breeders.

Spermatogenesis regeneration by transfected spermatogonial stem cells in infertile roosters through testicular transplantation.

Investigations pertaining to spermatogonial stem cells (SSCs) have led to the use of these cells in a variety of fields including infertility treatments, production of transgenic animals, and genome editing. The aim of the present study was to investigate the plausibility of regenerating spermatogenesis in infertile roosters by transplanting transfected SSCs into testes. Spermatogonial stem cells were isolated and cultured for seven days. Afterward, pDB2, a plasmid vector carrying a reporter gene, GFP, was transfected into the SSCs. Transfected SSCs were transplanted into the left testis of infertile roosters. Tissue samples from the recipients' testes were obtained six weeks after the transplantation and transplanted SSCs were observed in the basement membrane. After eight weeks, GFP-positive spermatozoa were observed in collected semen from the recipient roosters and GFP gene in spermatozoa was confirmed using PCR. The recipient roosters were mated with hens. Hatchlings were visually checked and their tissue samples were tested by PCR to identify transgenesis but both of them were negative. Overall, it seems that regeneration of spermatogenesis in roosters via transfected SSCs is possible but more studies are need to produce recombinant proteins by this way.

The effect of dietary supplementation of coenzyme Q10 on reproductive variables of cadmium-challenged male Japanese quails (Coturnix Japonica).

BACKGROUND: In recent decades, efforts to produce more efficient poultry products have increased due to its high demand. Meanwhile, some stressors have a negative impact on poultry efficiency and reproduction. Cadmium (Cd) is a toxic heavy metal with a high potential for inducing reactive oxygen species. On the other hand, coenzyme Q10 (CoQ10), with antioxidant properties, exerts a free radical-neutralizing effect on biological systems under stressful conditions. OBJECTIVES: This study aimed to determine the effect of dietary CoQ10 supplementation on reproductive variables of Cd-challenged male quails. METHODS: Two hundred and sixteen 42-day-old Japanese quails with a male-to-female ratio of 1:3 were randomly divided into three experimental groups (n = 72) and fed by experimental diets from 9 to 13 weeks of age (woa). Treatments included a negative control (NC): feeding basal diet; positive control (PC): feeding basal diet and Cd administration (1 mg per 100 g body weight at 10 and 11 woa); and CdQ10: dietary supplementation of CoQ10 (900 mg per kg diet) and Cd administration. At 10 and 13 woa, liver and testis, cloacal gland index, sera concentration of malondialdehyde (MDA) and testosterone, total antioxidant capacity (TAC), alanine aminotransferase (ALT), testicular histology, mRNA abundance of Hsp70 and fatty acid profile of testis, as well as hatchability and fertility, were measured. RESULTS: Liver and testis weights, cloacal gland index, serum concentration of testosterone, ALT, MDA, TAC, mRNA abundance of HSP70, hatchability, and fertility were not affected by the treatments. However, Cd administration decreased seminiferous tubule diameter and seminiferous epithelium thickness (SET) in the PC group compared to the NC group (p < 0.05). The proportion of saturated fatty acids (SFA) in testis tissue was increased, and the proportion of PUFA and n-3 to n-6 PUFA ratio was decreased in the PC group compared to the NC group (p < 0.05). In addition, CoQ10 supplementation ameliorated the effect of Cd on decreasing SFA and increasing n-3 to n-6 PUFA ratio proportions. CONCLUSIONS: In conclusion, Cd exerts several adverse effects on reproductive-associated variables; some, but not all, of them are mitigated by CoQ10 supplementation.

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

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). Duodenum VH increased in a linear and quadratic manner in response to increasing levels of acidifier. Egg weight, yolk diameter, jejunum CD, ileum CD, ileum VW, duodenum CD and jejunum VCR quadratically improved by grading levels of acidifier (P < 0.01). BW, albumen height, Haugh unit, ileum VH and ileum VCR were cubically enhanced (P < 0.05). Acidifier supplementation enhanced egg production, FCR, jejunum, ileum and cecum pH and VH, CD and VW of duodenum and jejunum, compared to the control group (P < 0.05); however, dietary acidifier did not affect egg mass, gizzard pH, ileum VH and bacterial count of the cecum (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.

Determining the Optimal Dosage of Lecithin Nanoliposome in Rooster Semen Freezing Medium and Fertility Potential.

Introduction: Lecithin nanoliposome (nano-LPO), with its cryoprotective properties, is considered to enhance the performance of a traditional semen cryoprotectant. Objective: To determine the optimal dose of lecithin nano-LPO added to the rooster semen extender. Materials and Methods: Semen samples collected weekly from eight broiler breeder roosters were mixed and aliquoted into five equal subsamples, during the five successive weeks. The subsamples were then diluted with a semen extender containing 0%, 0.5%, 1%, 1.5%, or 2% of lecithin nano-LPO. Post-thawed semen quality attributes, including sperm motility and velocity parameters, plasma membrane functionality, mitochondrial membrane potential (MMP), apoptosis-like changes, and fertility potential, were evaluated. Results: Total motility and velocity parameters, including curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity µm/s (VAP), straightness (STR), linearity (LIN), lateral head displacement (ALH), and wobble (WOB) were quadratically (p < 0.01) influenced by graded levels of lecithin nano-LPO, such that the highest values were obtained when 1% of lecithin nano-LPO was used. Treatments had no significant effect on plasma membrane functionality; however, MMP (p < 0.08) and percentages of live and dead spermatozoa (p < 0.05) quadratically responded to increasing levels of lecithin nano-LPO, where the best outcome was found when about 1% of lecithin nano-LPO was used in the semen extender. The percentage of apoptotic spermatozoa cubically responded to increasing levels of lecithin nano-LPO (p ≤ 0.07). No significant trend of fertility rate was found in response to addition of lecithin nano-LPO levels. Conclusions: Supplementing an extender with 1.10% of lecithin nano-LPO is shown to be the optimal dose associated with the most improvement in post-thawed rooster sperm velocity measurements.

The effect of dietary coenzyme Q10 supplementation on egg quality and liver histopathology of layer quails under cadmium challenge.

The demand for quail eggs has been increased over the last decade due to its beneficial nutritional quality characteristics; however, different nutritional and environmental stressors adversely impact the quality of the produced eggs. This study was conducted to investigate whether dietary supplementation of coenzyme Q10 (CoQ10) could mitigate the negative impact of cadmium (Cd) administration on egg quality and liver histopathology. A total of 162 six-week-old laying Japanese quail (Coturnix japonica) were randomly allotted into three experimental groups. Treatments were as follows: (1) negative control (NC): feeding basal diet; (2) positive control (PC): feeding basal diet and Cd administration; and (3) CdQ10: feeding basal diet supplemented with CoQ10 (900 mg/kg diet) and Cd administration. Cadmium (10 mg/kg BW) was subcutaneously administrated at 10 and 11 weeks of age (woa). Feed conversion ratio (FCR), egg production, egg mass, mortality rate, Cd residue in egg, liver histopathology, and some internal and external egg quality indices were evaluated. Administration of Cd increased FCR in the PC group, but supplementation of CoQ10 partially ameliorated the impact of Cd on FCR (p < 0.05). Cadmium administration decreased both egg production and egg mass; however, CoQ10 supplementation partially mitigated these adverse effects of Cd injection in the CdQ10 compared to the PC group (p < 0.05). Cadmium decreased eggshell thickness and Haugh unit in PC quail compared to both NC and CdQ10 quail (p < 0.05). Moreover, egg yolk colour intensity was enhanced by CoQ10, where a* and b* indices were higher in CdQ10 compared to PC (p < 0.05). In conclusion, the current results demonstrate the beneficial effects of dietary CoQ10 supplementation on liver histopathology and some egg quality indices of Cd-challenged quail.

Gene expression profile of placentomes and clinical parameters in the cows with retained placenta.

BACKGROUND: Retained placenta (RP) is a prevalent disorder in cattle with many health-related and economic costs for the farm owners. Its etiology has not been clarified yet and there is no definite therapy for this disorder. In this study we conducted RNA-seq, hematologic and histologic experiments to survey the causes of RP development. METHODS: Blood samples were collected from 4 RP and 3 healthy cows during periparturtion period for hematological assessments followed by placentome sampling within 30 min after parturition. Cows were grouped as RP and control in case the placenta was retained or otherwise expelled, respectively. Total RNA was extracted from placentome samples followed by RNA-sequencing. RESULTS: We showed 240 differentially expressed genes (DEGs) between the RP and control groups. Enrichment analyzes indicated immune system and lipid metabolism as prominent over- and under-represented pathways in RP cows, respectively. Hormonal assessments showed that estradiol-17ß (E2) was lower and cortisol tended to be higher in RP cows compared to controls at the day of parturition. Furthermore, histologic experiment showed that villi-crypt junctions remain tighter in RP cows compared to controls and the crypts layer seemed thicker in the placentome of RP cows. Complete blood cell (CBC) parameters were not significantly different between the two groups. CONCLUSION: Overall, DEGs derived from expression profiling and these genes contributed to enrichment of immune and lipid metabolism pathways. We suggested that E2 could be involved in development of RP and the concentrations of P4 and CBC counts periparturition might not be a determining factor.

Effects of dietary supplementation of different sources and levels of selenium on the semen quality and reproductive performance in aged broiler breeder roosters.

Fertility has a great impact on economic outcome in poultry sector. However, several physiological stressors such as aging adversely affected fertilization capacity and hatching quantity and quality. This study investigated the effect of dietary supplementation of different sources and levels of inorganic and organic selenium on the semen quality and reproductive performance of aged broiler breeder roosters. A total of thirty-six 50-wk-old Ross 308 roosters were randomly allocated to 6 groups and fed with different levels of organic and inorganic selenium. Treatments were included in the basal diet (control: CG), dietary supplementation of 0.15 (SeY0.15), 0.30 (SeY0.30), and 0.45 (SeY0.45) mg/kg selenium-enriched yeast (SeY), dietary supplementation of 0.30 mg/kg commercial organic selenium (Selemax), and dietary supplementation of 0.30 mg/kg sodium selenite (SS) as an inorganic source during 12 consecutive weeks. Ejaculated volume, semen quality attributes of the collected semen samples were evaluated every week. To assess fertility, hatchability and the hatched chick quality, the semen samples collected during last 2 wk of the trial were used to artificial insemination of hens. In order to measure seminiferous tube diameter and seminiferous epithelium thickness, testicular histology was also performed at the end of the experiment. Sperm motility, plasma membrane functionality and integrity, and ejaculation volume were higher in the SeY0.45 group compared to the other groups (P < 0.05). Fertility and hatchability rate as well as seminiferous epithelium thickness and seminiferous tube diameter were improved in the SeY0.45 compared with CG, SeY0.15 and SS groups (P < 0.05). Also hatchelling quality from roosters with SeY0.45 was higher than CG and SS groups (P < 0.05). No significant differences were noted in embryonic mortality between groups (P > 0.05). In conclusion, dietary supplementation of 0.45 mg SeY improved sperm quality and reproductive performance of aged broiler breeder roosters.

Supplementation of freezing medium with encapsulated or free glutathione during cryopreservation of bull sperm.

The objective was to compare effects of encapsulated or free glutathione (GSH) on the quality of frozen-thawed bull sperm. Ejaculates were collected via artificial vagina from six mature Holstein bulls once weekly for 6 weeks. All ejaculates had motility ≥70%, sperm concentration ≥1.0 × 109 /ml and ≤15% morphologically abnormal sperm. Each week, semen was pooled and diluted with lecithin-based extenders containing various concentrations of encapsulated (E0, E1, E2.5 and E5 mM) or free (F0, F1, F2.5 and F5 mM) GSH, with total glutathione content determined before and after cryopreservation. Total GSH in fresh semen was (mean+SEM) 4.8 ± 0.2 nmol/108 sperm, whereas in frozen-thawed semen of group F0 (control), it decreased to 1.4 ± 0.2 nmol/108 sperm, a 70.8% reduction (p < .05). In addition, total GSH in frozen-thawed semen from groups E2.5, E5 and F5 were 2.4 ± 0.2, 2.8 ± 0.2 and 1.8 ± 0.2 nmol/108 sperm, respectively (E5 versus. F0, p < .05). Compared to group F0, frozen-thawed sperm from group E2.5 had greater (p < .05) percentages of sperm that were viable (Annexin-V) (61.1 ± 1.8 versus. 71.1 ± 1.8) and that had cell membrane integrity (eosin-nigrosin) (64.5 ± 3.1 versus. 80.0 ± 3.1). Furthermore, frozen-thawed sperm from group E2.5 had the numerically highest total and progressive motility (CASA) and cell membrane functionality (HOS) and the lowest percentage of early apoptotic sperm (Annexin-V). However, acrosome membrane integrity (PSA) of E5 had the lowest mean (p < .05), whereas E2.5 caused a small nonsignificant decrease (69.1 ± 1.4%) compared to E0 and F0. In conclusion, 2.5 mM encapsulated GSH in semen extender significantly improved the quality of frozen-thawed bull sperm.

Effects of zinc dosage and particle size on gut morphology, tight junctions and TNF-α expression in broiler breeder hens.

This study was performed to evaluate the effects of different amounts and particle size of zinc oxide (ZnO) on villus height (VH), villus width (VW), crypt depth (CD) and VH to CD ratio (VH: CD), and expression of zonula occludens-1 (ZO-1), occludin (OC) and tumour necrosis factor-α (TNF-α) in broiler breeders. A total of 350 (Ross 308) broiler breeder hens of 54 weeks randomly assigned to seven treatments, included control basal diet (C) without added Zn, C+ 100, and 130 mg Zn per kg of diet from Large (L) (100-1000 nm) and Small (S) (<100 nm) particle size ZnO (LZnO100 and 130; SZnO100 and 130), C and SZnO100 challenged with lipopolysaccharide (C+LPS and SZnO100+LPS). Each diet was fed to five replicates consisting of ten birds each. The middle part of the duodenum, jejunum and ileum was used for morphological assessments. To assess the gene expression of ZO-1, OC and TNF-α in the jejunum samples were excised. Results showed that the supplementing 130 ppm SZnO increased VH:CD in the duodenum (p < 0.05). VW in the duodenum and all the evaluated morphometric indices in jejunum and ileum were not affected by the dietary treatment (p > 0.05). ZO-1 mRNA abundance in C+LPS group compared to SZnO100+LPS group was significantly decreased and increased by LPS and SZnO100 respectively. The SZnO-100 increased OC gene expression in compare to C+LPS group. The expression of TNF-α in C+LPS treatment was higher than other groups (p < 0.05). The lowest and the highest litter moisture and foot-pad dermatitis (FPD) were observed in LZnO-130 and C treatments respectively (p < 0.05). Improving the physical properties of ZnO affect on VH:CD. Broiler breeder diet with ZnO enhance ZO-1, OC and mitigate TNF-α gene expression in jejunum maintenance of gut health in broiler breeders.

Effects of coenzyme Q10 on reproductive performance of laying Japanese quail (Coturnix japonica) under cadmium challenge.

Japanese quail is an increasingly important bird of economic importance for commercial egg and meat production, particularly in developing countries. There is a need for research aimed at improving efficiency of these birds during stressful challenges, such as oxidative stress. Coenzyme Q10 (CoQ10), a highly functional antioxidant, protects cells against oxidative stress. This study was conducted to determine the effects of CoQ10 on reproductive performance of Japanese quail under cadmium (Cd) challenge. A total of 216 six-wk-old Japanese quail were randomly allocated into 3 groups for an 8 wk experimental trial. The treatments include a negative control (NC): feeding basal diet; a positive control (PC): feeding basal diet and cadmium administration (1 mg/100 g BW, at 10 and 11 wk of age), and (CdQ10): feeding CoQ10 supplemented (900 mg/kg diet) basal diet and Cd administration. At 11 and 13 wk of age, egg production, body weight, mortality, oviduct, and ovarian biometry, were recorded. Histology and histopathology of isthmus and magnum, fertility, hatchability, hatchling quality, and HSP70 mRNA transcript abundance in the utero-vaginal junction (UVJ) were evaluated. Positive control and CdQ10 group had no significant effect on live body weight, stroma weight, follicle size, hatchability, and fertility; however, Cd administration increased (P < 0.01) mortality rate in the PC group compared to the NC and CdQ10 groups. CdQ10 quail produced more eggs and had a higher hatchling quality compared to the PC group (P < 0.01). The thickness and height of isthmus and magnum folds in the CdQ10 group was increased compared to the PC group (P < 0.01) and overall oviduct weight was increased with CoQ10 supplementation (P < 0.01). Compared to PC, the CdQ10 group had a reduction in infiltration of inflammatory cells. Relative abundance of HSP70 mRNA in UVJ was influenced by interactive effect of treatment × time (P < 0.05). In conclusion, dietary supplementation of CoQ10 showed beneficial effects on some reproduction characteristics of female Japanese quail under Cd-induced oxidative stress.

The effects of varying concentrations of glutathione and trehalose in improving microscopic and oxidative stress parameters in Turkey semen during liquid storage at 5 °C.

Since turkey reproduction is mainly through artificial insemination, short-term preservation of turkey semen is one of the most important issues in turkey reproduction management. The present study investigates the effects of glutathione (GSH) and trehalose on lipid peroxidation degree and turkey semen quality while being stored at 5 °C for 72 h. To this end, semen samples were collected from 20 turkeys with a weekly frequency for 12 weeks. A glucose-based extender was used to dilute the pooled semen. It was divided into seven equal parts with varying levels of glutathione [0.5, 1 and 2 mM), trehalose [50, 75 and 100] and control [extender without antioxidant]. Subsequently, the divided semen samples were stored at 5 °C for 72 h. Several sperm parameters such as motility and motion parameters, plasma membrane integrity (PMI), plasma membrane functionality, DNA integrity, and oxidative parameters were assessed following storage for 0, 24, 48, and 72 h. The obtained results indicated an improvement in the plasma membrane functionality and DNA integrity, along with the percentages of PMI in GSH-2 mM group in comparison to the control group following storage at 5 °C for 72 h (P ≤ 0.05). It is also notable that the 2 and 1 mM concentrations of GSH increased the spermatozoa motility and motion parameters in comparison to the control group, respectively (P ≤ 0.05). The study results indicated that GSH-2, 1 mM and trehalose- 100 mM concentrations reduced lipid peroxidase levels and increased total antioxidant activity, catalase, superoxide dismutase, and glutathione peroxidase in comparison to the control group (P ≤ 0.05). Our study's data show that improvement of semen parameters and oxidative stress parameters of turkey semen can be improved by glutathione at 2 and 1 mM and trehalose at 75 mM while storing it 5 °C.

Effects of different ultrastructures of lecithin on cryosurvival of goat spermatozoa.

This study was to evaluate the effects of two different ultrastructures of lecithin including nanoparticles (NPE mostly nanomicelles) and lecithin nanoliposome (NLE) with egg yolk extender (EYE) on goat sperm cryopreservation. Semen samples were collected from 6 goats, then pooled, diluted and then frozen. Motility and motion parameters, plasma membrane integrity and functionality, morphology, apoptosis status (Annexin V-PI), acrosome integrity, DNA fragmentation and in vitro fertilisation were assessed. Total motility and most motion parameters were higher in EYE (p < .05) compared with the two lecithin extenders, while there were no significant differences between NLE and NPE. NLE and NPE had higher values for viable spermatozoa (Annexin V-PI) (p < .05) compared with EYE. The highest value for dead spermatozoa was observed in EYE (p = .08). A higher percentage of DNA fragmentation (p < .05) was detected in EYE compared with NPE. Plasma membrane integrity and functionality, morphology, acrosome integrity and fertility of spermatozoa indicated no significant differences between extenders. Data suggested that ultrastructural changes of lecithin (micelles versus. liposome) could not improve the sperm cryosurvival of goat spermatozoa. Moreover, we cannot also claim that lecithin-based diluent supplies better protection compared with the egg yolk in goat.

Conjugated linoleic acid improves meiotic spindle morphology and developmental competence of heat-stressed bovine oocyte.

This study was conducted to elucidate the effects of introducing conjugated linoleic acid (CLA) on meiotic spindle organization of heat stressed (HS) matured oocytes and the resulting blastocysts DNA methylation as well as the expression of the genes involved in DNA methylation (DNMT3a, DNMT3b and DNMT1). Immature bovine oocytes were cultured at 41 °C for the first 12 h and 38.5 °C for the second 12 h of maturation time in the presence of 0 and 50 µM of CLA (HS and HS + CLA groups, respectively). A group of oocytes cultured in medium with no CLA supplementation at normal temperature (38.5 °C for 24 h) was considered as negative control (C). Percentage of normal spindle, and cleavage and blastocyst rates were significantly decreased in the HS group compared to the C group (P < 0.05). The global DNA methylation and expression level of DNMT3a gene were increased in HS group compared to the C groups (P < 0.05), while the expression level of DNMT3b was decreased. The CLA supplementation improved the percentage of normal microtubules shape in MII oocytes as well as the developmental competence in the HS + CLA group compared to the HS group (P < 0.05). However, global DNA methylation and expression level of DNMT3a/b were not ameliorated by CLA supplementation (P > 0.05). Based on the obtained results, CLA proved to be capable of improving the oocyte developmental competence as well as decreased the aberrant spindle organization of heat-stressed oocytes and it would not cause epigenetic alteration in the obtained blastocysts.

The effect of dietary organic selenium on reproductive performance of broiler breeder roosters under dexamethasone induced stress.

Stress has deleterious impact on semen quality and fertility of roosters. This study was investigated to know whether dietary supplementation of organic selenium (oSe) could improve semen quality and fertility of male broiler breeder under dexamethasone (Dexa) induced stress. Forty broiler breeder roosters (64 week of age) were randomly allotted to four groups (10 roosters/group) and fed a standard diet supplemented with different levels of oSe during 10 successive weeks of the experimental period. To induce stress, the birds received injections of 2 mg/kg BW of Dexa during weeks 5 and 6 of the experiment, in one-day-intervals manner. The roosters were not treated with Dexa and oSe (negative control; NC), or treated with Dexa and different levels of oSe including 0 (positive control; PC), 0.30 (Se30+Dexa) or 0.45 (Se45+Dexa) mg/kg diet. Body weight was measured weekly and semen quality parameters and fertility were evaluated every two weeks. Except for seminal volume and total sperm production which was not affected by the treatment, body weight, semen quality parameters, total antioxidant capacity (TAC) and malondialdehyde (MDA) in seminal plasma were influenced by interactive effect of treatment and time (P < 0.05). Dexamethasone injection adversely affected semen quality parameters (semen concentration, motility and plasma membrane integrity) in PC group compared to NC group (P < 0.05); however, dietary supplementation of oSe ameliorated these negative impacts in Dexa-treated roosters (P < 0.05). Fertility was also improved by dietary supplementation of oSe compared to control groups (P < 0.05). These results indicate that although induction of stress have negative effects on rooster semen quality parameters, dietary inclusion of oSe may exert beneficial impact on mitigating the harmful effects of stress on semen quality and fertility rate of broiler breeder roosters.

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), whereas semen volume, subjective motility, and total sperm output were not affected (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.

Distribution and size of lipid droplets in oocytes recovered from young lamb and adult ovine ovaries.

This study evaluated the distribution and size of lipid droplets (LDs) in oocytes recovered from young and adult ovine ovaries. Collected oocytes were categorised on the basis of their major diameter (small (SO), 70-90 µm; medium (MO), >90-110 µm; large (LO), >110-130µm) and were stained with Nile red to detect LDs. In adult and young oocytes, a diffuse pattern distribution of LDs was dominant in all classes except adult LO and young SO and LO. Larger LDs (i.e. >3µm) were mostly present in young SO and LO, whereas smaller LDs (1-3µm) were detected in the other adult and young oocyte categories.

Dietary omega-3 fatty acids from linseed oil improve quality of post-thaw but not fresh sperm in Holstein bulls.

Docosahexaenoic acid (DHA), a member of the n-3 fatty acid family present in fish oil, has several positive effects on bovine sperm, including membrane integrity, motility and viability, as well as cold sensitivity. Our objective was to investigate effects of varying amounts of omega-3 fatty acids from linseed oil, administered orally, on quality of fresh and frozen-thawed bull sperm. Twenty fertile Holstein bulls (874 ± 45.38 kg) were randomly and equally assigned to four groups and received encapsulated (rumen-protected) fats for 12 weeks, as follows: group P, 300 g palm oil; group Pl, 200 g palm oil + 100 g linseed oil; group pL, 100 g palm oil + 200 g linseed oil; and group L, 300 g linseed oil. Sperm quality of fresh and frozen-thawed semen was evaluated by routine assays including sperm motion characteristics (CASA), membrane integrity (eosin-nigrosin), membrane activity (hypo-osmotic swelling test; HOST) and malondialdehyde (MDA) content. There were no significant differences among groups in semen volume, sperm concentration or sperm quality parameters in fresh semen. However, after freezing-thawing, total and progressive motility in group P (59.61 ± 1.95 and 40.19 ± 2.48%, respectively; LSM ± SEM) were lower (P < 0.05) than in groups Pl (66.06 ± 1.95 and 47.53 ± 2.48%), pL (65.67 ± 1.95 and 47.48 ± 2.48%) and L (65.36 ± 1.95 and 47.62 ± 2.48)%, with no significant differences among the latter three groups. Furthermore, membrane integrity (eosin-nigrosin) and activity (HOST) were lower (P < 0.05) in group P (55.79 ± 2.15 and 42.19 ± 2.17%) compared to groups Pl (62.73 ± 2.15 and 48.93 ± 2.17%), pL (64.06 ± 2.15 and 50.01 ± 2.17%) and L (64.47 ± 2.15 and 49.68 ± 2.17%), with no significant differences among the latter three. Furthermore, there were more (P < 0.05) morphologically abnormal sperm in group P (25.99 ± 1.62%) than in groups Pl, PL and L (21.55 ± 1.62, 21.69 ± 1.62 and 20.90 ± 1.62%). In conclusion, feeding Holstein bulls 100-300 g linseed oil daily improved sperm cryotolerance.