Effect of varying amounts of linseed oil or saturated fatty acids around insemination on reproductive and blood parameters of ewes.

The current study was designed to evaluate the effect of sequential low and high dietary linseed oil (LO; as omega-3 enriched fatty acid; FA) before and post insemination, respectively, on different plasma variables of ewes. Fat-tailed Qezel ewes were assigned randomly to be fed a diet enriched with 3% LO (n = 30) or the saturated FA (SFA; n = 30) three weeks before insemination (Day 0). The lipogenic diet supplemented with 6% LO or SFA was fed after insemination until Day +21. The control ewes were fed an isocaloric and isonitrogenous diet with no additional FA during the study. Estrus was synchronized by inserting a vaginal sponge (Spongavet®) for 12 days + 500 IU equine chorionic gonadotropin (eCG; Gonaser®), and ewes were inseminated via laparoscopic approach 56-59 h after eCG injection. The size of ovarian structures was assessed by transvaginal ultrasonography at -21, -14, -2, 0, and +10 days. Blood samples were collected weekly to measure the plasma's different biochemical variables and FA profile. Treatment did not affect the amounts of glucose, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, interleukin-10, interleukin-2, and non-esterified FA (p > 0.05). Conversely, concentrations of triglyceride, cholesterol, tumor necrosis factor-alpha, and insulin-like growth factor-1 were higher in SFA-fed ewes relative to control animals (p < 0.05). LO feeding resulted in greater amounts of n-3 FA isomers in plasma, while higher amounts of stearic acid were detected in SFA fed group 0 and +21 (p < 0.05). The number of ovarian follicles and corpora lutea also were not affected by treatment. Other reproductive variables were not affected by treatment except for the reproductive rate. It seems that LO or SFA feeding of fat-tailed ewes peri-insemination period was not superior to the isocaloric non-additional fat diet provided for the control group during the non-breeding season.

Timing of ovulation in the fat-tailed Qezel ewes after synchronization with vaginal devices containing endogenous or exogenous synthetic progestogens during out of the breeding season.

BACKGROUND: Wide range of ovulation distribution is the main restricting factor in establishing the pregnancy following oestrus synchronization (ES) and fixed time insemination (FTI) in sheep. OBJECTIVES: Determining the ovulation time (OVT) following ES with two different vaginal devices, its relation to progesterone and conception upon FTI with frozen/thawed semen. METHODS: Oestrus was synchronized using either controlled internal drug release (CIDR) (ewe, n = 6; ewe lamb, n = 5) or vaginal sponge (ewe, n = 6; ewe lamb, n = 5) insertion for 12 days, plus Equine chorionic gonadotropin (eCG) at devices removal (Day 0). Sizes of the ovarian follicles were measured using transvaginal probe at -12, 0 and 30-33, 53 h and continued every 3-4 h until 75 h after eCG treatment. Serum progesterone amounts were measured at -12, 0, +2 and +11. Laparoscopic FTI was done at 60.5 ± 0.5 h. RESULTS: The CIDR-treated group initiated and completed ovulations earlier compared to sponge-treated females (median: 64 vs. 71 h; p < 0.05). Ewe lambs were ovulated earlier compared to ewes in the sponge-treated group (66.71 vs. 71.5; p = 0.017). Mean sizes of ovulatory follicles and corpora lutea were not affected by device types. Higher amounts of progesterone were observed in CIDR group compared to sponge-treated group at device removal (2.68 ± 0.12 vs. 0.30 ± 0.01 ng/mL; p < 0.001). The conception was confirmed in 2/10, and 5/11 females of sponge and CIDR-treated females, respectively. CONCLUSIONS: Types of progestogens influence the OVT, and consequently the result of FTI with frozen/thawed semen. The optimum timespan for FTI should be chosen according to the device types during non-breeding season.

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.

Influence of trans-ferulic acid on the quality of ram semen upon cold preservation.

BACKGROUND: Due to lower antioxidant capacity and higher amounts of polyunsaturated fatty acids, ram spermatozoa are very susceptible during cooling process. OBJECTIVES: The objective was to examine the effect of the trans-ferulic acid (t-FA) on the ram semen during liquid preservation. METHODS: Semen samples were collected from the Qezel rams, pooled, and extended with the Tris-based diluent. Pooled samples enriched with different amounts of the t-FA (0, 2.5, 5, 10, and 25 mM) and preserved at 4°C for 72 h. Spermatozoa's kinematics, membrane functionality, and viability were assessed by CASA system, hypoosmotic swelling test, and eosin-nigrosin staining, respectively. Moreover, biochemical parameters were measured at 0, 24, 48, and 72 h. RESULTS: Results showed that 5 and 10 mM t-FA improved forward progressive motility (FPM) and curvilinear velocity compared to the other groups at 72 h (p < 0.05). Samples treated with 25 mM t-FA showed the lowest total motility, FPM, and viability at 24, 48, and 72 h of storage (p < 0.05). Higher total antioxidant activity levels were observed in the 10 mM t-FA-treated group compared to the negative control at 72 h (p < 0.05). Treatment with 25 mM t-FA increased malondialdehyde amounts and decreased superoxide dismutase activity compared to other groups at the final time assessment (p < 0.05). Nitrate-nitrite and lipid hydroperoxides values were not affected by treatment. CONCLUSIONS: The current study indicates the positive and negative influences of different concentrations of t-FA on the ram semen upon cold storage.

Evaluation of Chicken Egg Yolk Plasma and Low-Density Lipoprotein Alone or Enriched with Ewe or Cow Skim Milk in Tris-Citric Acid-Based Diluent for Cryostorage of Ram Semen.

The main purpose of the current study was to find suitable and optimum levels of protectants among chicken egg yolk plasma (CEYP) and low-density lipoproteins (LDLs), alone or supplemented with ewe or cow skim milk, for cryopreservation of ram semen. In Experiments 1 and 2, the CEYP (28%) freezing extender was enriched with ewe or cow milk (2.5%, 5%, 10%, or 20%; v/v), respectively. In Experiments 3 and 4, the semen extender was prepared by varying the amounts of fresh or lyophilized LDL (lyo-LDL), respectively. Finally, ewe or cow skim milk was added to the freshly extracted LDL extender and the quality of frozen-thawed semen was examined (Experiments 5 and 6). Kinematics of spermatozoa (assessed using a computer-assisted sperm analysis system), viability, functionality of the plasma membrane, and levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) were evaluated. Results revealed that addition of ewe or cow skim milk (5%, 10%, or 20%; v/v) to the CEYP diluent enhanced kinematics, viability, and membrane integrity of spermatozoa compared with the control (p < 0.05). Moreover, fresh LDL diluent was more effective than lyo-LDL in the cryosurvival of ram spermatozoa. In addition, enrichment of fresh LDL diluent with ewe or cow skim milk improved different variables of spermatozoa compared with the control (p < 0.05). Levels of MDA and TAC were not affected by adding ewe or cow milk to the diluents (p > 0.05). In conclusion, enrichment of fresh LDL extenders with ewe or cow milk also is proposed as an approach to preserve ram semen quality against cold shock and cryodamage injuries.

Ameliorative role of trans-ferulic acid on induced oxidative toxicity of rooster semen by ß-cyfluthrin during low temperature liquid storage.

Current study was designed to evaluate the effects of ß-cyfluthrin, as a toxicant substance, and trans-ferulic acid (trans-FA), as a protective agent, on different parameters of rooster semen upon liquid storage. For this purpose, semen samples of roosters (Ross 308, n = 10, 32-wk-old) were collected twice a week. Good quality samples (≥70% progressive motion) were diluted, pooled and then divided for the purposes of the study. In the first experiment, motility of spermatozoa was evaluated following exposure to different concentrations of ß-cyfluthrin (1, 2.5, 5, 10, 20, 40, and 80 µM) at 0, 24, and 48 h of storage. In the second experiment, constant doses of ß-cyfluthrin (10 µM) alone or in combination with trans-FA (10, 25 mM) were assessed on motility and viability of spermatozoa at 0, 24, and 48 h time points. Moreover, amounts of malondialdehyde (MDA), total antioxidant capacity (TAC), total nitrate-nitrite, total hydroperoxide (HPO), and activity of superoxide dismutase (SOD) were evaluated in the homogenate of spermatozoa-diluent at studied time points. Results of the first experiment showed that amounts of ß-cyfluthrin greater than 5 µM, significantly reduced the motility of spermatozoa at 24 and 48 h of storage (P < 0.05). The second experiment demonstrated that, trans-FA especially at 10, 25 mM doses restored the motility and viability of spermatozoa compared to ß-cyfluthrin treated group (P < 0.05). Amounts of MDA (10, 25 mM), hydroperoxide (10, 25, and 50 mM), and nitrate-nitrite (10, 25, and 50 mM) were lower and TAC (10 and 25 mM) were greater in trans-FA + ß-cyfluthrin treated groups compared to ß-cyfluthrin alone treated samples (P < 0.05). However, activity of SOD did not show significant changes by the treatment (P > 0.05). It seems that trans-FA could ameliorate toxic effect of ß-cyfluthrin via reduction of peroxidative (as evident by measurement of MDA) and nitrosative (as evident by measurement of nitrate-nitrite) reactions over cold preservation of rooster semen.

Comparison of different carrier-compounds and varying concentrations of oleic acid on freezing tolerance of ram spermatozoa in tris-citric acid-egg yolk plasma semen diluent.

The current study was conducted to determine the optimal concentration carrier-compound for oleic acid (OA) among dimethyl sulfoxide (DMSO), liposome and ß-cyclodextrin on ram spermatozoa cryosurvival. The preliminary experiment was designed to ascertain the optimal concentration of egg yolk plasma. In Experiment 1, semen was placed in a diluent containing different concentrations of OA dissolved in DMSO (0.125, 0.25, 0.50, 1, 2, 4 and 8 mM). In Experiments 2 and 3, effects of liposome loaded-OA and ß-cyclodextrin-OA complexes (0.25, 0.50, 1 and 2 mM) on semen cryopreservation were evaluated. In Experiment 4, optimal concentrations of OA were determined, based on results from previous experiments. Spermatozoa viability, kinematics, plasma membrane integrity, malondialdehyde, superoxide dismutase activity and total antioxidant status of samples were evaluated. Results indicated varying concentrations of OA had different effects on sperm kinematics, viability and membrane functionality after freezing/thawing (P < 0.05). In addition, inclusion of OA in liposomes or combinations with ß-cyclodextrin resulted in greater values for spermatozoa motion variables compared with DMSO dissolved-OA (P < 0.05). Inclusions of OA at 0.25 and 0.50 mM led to a reduction in amounts of lipid peroxidation when there was inclusion of liposome and ß-cyclodextrin as carrier-compounds (P < 0.05). Activity of SOD was similar with inclusion of different concentrations of OA or carrier-compounds, but total antioxidant capacity was affected by OA concentration and carrier-compound type (P < 0.05). The results highlight the importance of carrier-compound type and concentrations of OA on ram spermatozoa during cryopreservation.

Quercetin attenuates H2 O2 -induced toxicity of rooster semen during liquid storage at 4°C.

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H2 O2 (40 µM) and combination groups which incubated with constant dose of H2 O2 (40 µM) plus various levels of quercetin (20, 40 and 80 µM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 µmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 µmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 µmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co-administration of quercetin (especially at 40 and 80 µM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide-mediated damage during cold liquid storage of rooster semen.

Influence of idebenone on ram semen quality stored at 4°C.

The current study was designed to investigate the effect of idebenone (Id), an antioxidant on ram semen quality. Semen samples were collected, pooled and diluted in a Tris-based extender supplemented with 0, 1, 2, 4 and 8 µM idebenone. Computer-assisted sperm analysis was used to evaluate spermatozoa kinematics. Sperm viability and membrane functionality were assessed respectively, by eosin-nigrosin staining and HOS test. Biochemical assays were carried out to measure different metabolites in spermatozoa and medium at 0, 24, 48 and 72 hr. Total and forward progressive motility were greater in 1, 2 and 4 µM idebenone treated groups compared to control at 24, 48 and 72 hr time points (p < 0.05). Semen supplementation with Id significantly increased viability and functionality of spermatozoa membrane during storage (p < 0.05). Lower amounts of lipid hydroperoxides in medium and spermatozoa were observed in Id-treated groups compared to control one at 24 and 48 hr of storage (p < 0.05). Medium and spermatozoa amounts of malondialdehyde and nitric oxide were less in Id 4 µM group compared to the control at 72 hr (p < 0.05). Total antioxidant capacity values and superoxide dismutase activity of spermatozoa and medium were greater in 2 and 4 µM idebenone treated groups in comparison with the control at 72 hr (p < 0.05). Results of the current study indicated that ram semen supplementation with Id at 4 µM level improved quality by ameliorating nitrosative and peroxidative stress, hence could be considered as an antioxidant additive during storage at 4°C.

Evaluation of linoleic acid on lipid peroxidative/antioxidative parameters, motility and viability of rooster spermatozoa during cold storage.

Sustainable reduction in semen quality due to detrimental effects of primary and secondary peroxidative products was occurred during liquid storage. The objective of the current experiment was to explore the influence of bovine serum albumin conjugated linoleic acid (LA) on the rooster spermatozoa routine tests and lipid peroxidative/antioxidative levels stored at 4 °C over 48 h. For this purpose, collected ejaculates (≥ 80% progressive motile spermatozoa) pooled and extended with the phosphate buffer medium without (control) or enriched with different amounts of LA (0.125, 0.25 or 0.50 mM). Contents of total antioxidant status (TAS) and thiobarbituric reactive substances (TBARS) were measured separately in the medium and spermatozoa, as well as percent of viability and motility at 0, 24 and 48 h intervals. Viability was not affected by treatment during the study intervals (P > 0.05). While, higher motility was recorded in LA 0.50 mM group (77.98 ± 1.89 and 57.02 ± 2.45) compared to the control group (68.78 ± 1.29 and 45.09 ± 1.86) at 24 and 48 h, respectively (P < 0.03). Amounts of TBARS in medium and spermatozoa were lower in LA 0.25 and 0.50 groups compared to the control at 48 h (P < 0.01). Moreover, TAS levels of medium and spermatozoa were lower in control samples compared to LA treated groups at 48 h (P < 0.03). Because of the ability of the LA to lowering the quantities of lipid peroxidation index and improving motility especially at 0.5 mM levels, it can be proposed as an additive during liquid storage of rooster semen.

Kinetin improves motility, viability and antioxidative parameters of ram semen during storage at refrigerator temperature.

The present experiment was conducted to evaluate the effect of kinetin on ram semen quality during cold storage. Ejaculates were collected using an artificial vagina from five Qezel rams. Ejaculates which met the criteria (volume of 0.75-2 ml; minimum spermatozoa concentration of 2.5 × 109 spermatozoa/ml and forward progressive motility of 80%), were pooled, diluted with extender without kinetin (control) or enriched with 25 (K 25), 50 (K 50), 100 (K 100) and 200 (K 200) µM kinetin at a final concentration of 500 × 106 spermatozoa per mL. Spermatozoa motion characteristics were evaluated by computer-assisted sperm analysis. In addition, percent of viability (spermatozoa showing no color was considered to be alive) and spermatozoa with intact plasma membrane (spermatozoa with curled/swollen tail was considered healthy) were determined. Moreover, amounts of malondialdehyde (MDA), total antioxidant activity (AOA), nitric oxide (NO) and superoxide dismutase (SOD) activity were determined in the seminal plasma and spermatozoa at 0, 24, 48 and 72 h of storage. Higher percent of total and forward progressive motility was observed in K 25, K 50 and K 100 groups compared to control group at 72 h of storage (P < 0.001). Moreover, K 25 (78.61 ± 1.11%), K 50 (82.46 ± 1.08%) and K 100 (82.96 ± 1.49%) groups showed higher percentage of viability compared to control (72.38 ± 1.49%) at 72 h of storage (P < 0.05). Semen enrichment with kinetin resulted in the higher percent of intact plasma membrane of spermatozoa at 48 and 72 h (P < 0.001). Amounts of MDA were lower and amounts of AOA were higher in K 50 and K 100 groups compared to control at 48 and 72 h (P < 0.05). There were no significant differences in NO levels and SOD activities of seminal plasma and spermatozoa among groups during the experiment. The present experiment revealed that kinetin at proper concentration could enhances spermatozoa kinematics, viability, spermatozoa plasma membrane functionality and amounts of AOA and reduces the level of lipid peroxidation during chilled storage of ram semen.

Palmitoleate enhances quality of rooster semen during chilled storage.

The practice of artificial insemination is widely utilized in poultry; and this requires a broad use of semen storage techniques to prevent the reduction of fertilizing ability of stored semen. The antioxidant activity of palmitoleic acid with in vitro experiments has been shown. The present study was designed to evaluate the effect of palmitoleic acid on the quality of rooster semen stored at 4C. Semen was collected from ten roosters twice a week. Ejaculates with greater than 80% forward spermatozoa motility were pooled and after dilution semen was enriched with 0 (control), 0.125 (P 0.125), 0.25 (P 0.25), 0.5 (P 0.5) and 1 (P 1) millimolar palmitoleate. Forward spermatozoa progressive motility and viability, as well as amounts of malondialdehyde (MDA) and total antioxidant activity (AOA) were evaluated in seminal plasma and spermatozoa at 0, 24 and 48h of storage. Motility was 78.5±2.21, 77.5±1.04, and 69.5±2.32% at 24h and 58.66±1.35, 49.33±1.36 and 43.00±2.08% at 48h in P 0.125, P 0.25 and control, respectively (P<0.02). There were no significant differences in amount of MDA in the seminal plasma among groups, while the amounts of MDA in spermatozoa were less in the P 0.125, P 0.25 and P 0.5 groups compared to the control group at 24 and 48h of storage (P<0.002). Total amounts of AOA in seminal plasma were greater in palmitoleate treatment groups than the control at 24 and 48h (P<0.01). Moreover, palmitoleate treatment groups had greater values of total AOA in spermatozoa compared to the control group at 24 and 48h of storage (P<0.05). In conclusion, enrichment of rooster semen with small doses of palmitoleate has beneficial effects on the semen quality during cold storage.

Treatment of clinical endometritis in dairy cows by previously used controlled internal drug release devices.

Postpartum endometritis is considered as one of the diseases that lead to a potential profit reduction in dairy cows. The aims of the present study were to promote follicle growth by a previously used controlled internal drug release (CIDR) device and to evaluate its effect on the likelihood of recovery and the reproductive performance of clinical endometritis (CE) cows. Endometritis was diagnosed using ultrasonographic examination at 31 ± 3 (Day 0 of the experiment) days in milk, and CE cows were included in one of the three experimental groups according to the presence of a CL on their ovaries. Cows without CL on their ovaries received a reused CIDR device, which was previously used for 14 days (CIDR-14, n = 108), or PGF2α (PG-1, n = 112) on Day 0. In the third group, those with CL on their ovaries received PGF2α (PG-2, n = 107) at the same time. Ovarian structures, serum estradiol and progesterone concentrations were measured on Days 0, 7, and 14. Controlled internal drug release devices were removed, and response to treatment was evaluated in all treated cows on Day 14. Diameters of ovarian follicles were 11.61 ± 0.50, 12.46 ± 0.25, and 18.36 ± 0.60 mm on Day 7 and 11.63 ± 0.58, 14.35 ± 0.40, and 21.96 ± 0.77 mm on Day 14 in PG-1, PG-2, and CIDR-14 cows, respectively (P < 0.05). Serum estradiol concentrations were higher in CIDR-14 cows (141.17 ± 1.04 pg/mL) than in PG-1 (116.85 ± 1.05 pg/mL) and PG-2 (119.10 ± 1.05 pg/mL) cows on Day 7 (P < 0.05). Higher progesterone concentrations were observed in PG-2 cows than in PG-1 and CIDR-14 cows on Days 0, 7, and 14 (P < 0.001). The likelihood of clinical cure was 54.46%, 62.61%, and 64.81% in PG-1, PG-2, and CIDR-14 cows, respectively (P = 0.11). First-service conception risk, days to the first service, calving to conception interval, proportion of cows bred and pregnant by 120 days in milk did not differ among the treated groups (P > 0.05). The cumulative pregnancy risk was lower in PG-1 (77.67%) cows than in CIDR-14 (87.07%) and PG-2 (87.85%) cows (P = 0.02). In conclusion, reused CIDR would be contributed to the treatment of CE by promotion of follicle growth and induction of sustainable sources of endogenic estrogen secreted by the dominant follicle.