Reducing oxidative stress by κ-carrageenan and C60HyFn: The post-thaw quality and antioxidant status of Azari water buffalo bull semen.

Azeri water buffalo is a species of great interest due to the high quality of its products such as milk. Due to the decreasing trend of its number and risk of extinction in the future, our attention is directed towards ensuring the preservation of its genetic reserves by keeping its sperm. Using antioxidants in semen extender is one of the ways to reduce the detrimental effects of freezing process on post-thawed quality of spermatozoa. This study was conducted to determine the effect of κ-carrageenan (k-CRG) and C60HyFn supplemented semen extender on the quality of post-thawed Azari water buffalo spermatozoa. A total of 30 semen samples were obtained from three buffaloes using an artificial vagina (twice a week for five weeks = 10 replicates). The samples (n = 3) from each replicate were pooled and divided into equal aliquots to prepare 14 extender groups, including control (C), k-0.2, K-0.4, K-0.6, K-0.8 (containing 0.2, 0.4, 0.8 mg K-CRG/mL, respectively), C-0.1, C-0.2, C-0.4, C-0.8, C-1, C-5, C-10, C-20, and C-40 (containing 0.1, 0.2, 0.4, 0.6, 0.8, 1, 5, 10, 20, 40 µM C60HyFn, respectively), and then frozen. After thawing, motility and velocity parameters, plasma membrane integrity (PMI) and functionality (PMF), DNA damage, Hypo-osmotic swelling (HOS) test, malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase glutathione activities and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging were evaluated. In vivo fertility was compared between k-0.6, C-1 and control groups. 60 buffalo were inseminated 24 h after the onset of estrus. The diagnosis of pregnancy was performed rectally at least 60 days after fertilization. Total and progressive motility and velocity parameters were improved by k-0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups) compared to the other groups. Plasma membranes integrity and PMF were improved by k-0.4, k-0.6, C-0.4, C-0.8, C-1, C-5, and C-10 groups compared to other groups, while in terms of sperm DNA damage K-0.4, K-0.6, K-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups showed better results compared to the control group. The evidence also showed that k- 0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve TAC, and decrease MDA levels. Also, k-0.4, k-0.6, k-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve GPx, CAT, and GSH levels, but no significant difference was found regarding SOD compared to the other groups. DPPH scavengers were tested by K-0.6, K-0.8 and C-1, C-5, C-10, C-0.8, C-0.4 and C-0.2 groups and compared to other groups improved. The fertility rate [70% (14/20)] was higher in C-1 than other groups. To conclude that k-CRG and C60HyFn supplementation can increase the quality parameters of cryopreserved buffalo semen after thawing and that 1 M C60HyFn can increase in vivo fertility of buffalo semen.

Effect of Monovalent Copper Oxide and Potentiated Zinc Oxide on Growth Performance and Gut Morphology of Broiler Chickens Challenged with Coccidiosis.

An experiment was conducted to evaluate the effect of copper oxide (Cu2O) and potentiated zinc oxide (ZnO) on performance, intestinal morphology, oocyst excretion, coccidial lesion scores, and antioxidant properties in broilers during an Eimeria spp. challenge. A total of 288 1-day-old male broiler chickens (Ross 308) were divided into 18 treatments. Treatments included three levels of Cu (0, 15, or 150 mg/kg) from Cu2O and three levels of Zn (0, 80, or 160 mg/kg) from potentiated ZnO which were added to the basal diet and fed to broilers with or without challenge, using a completely randomized design in a factorial arrangement for 42 days. Live body weight, feed intake, mortality, and the cause of death were recorded weekly and histomorphology of jejunum was measured at the end of the experiment. Results showed that birds fed Cu and Zn linearly decreased (P < 0.0001) oocyst shedding. The number of excreted oocysts was reduced eight times in broilers fed a diet containing 150 mg/kg copper from Cu2O and 160 mg/kg zinc from potentiated ZnO, compared to the infected group without Cu and Zn supplementation (P < 0.0001). Microscopic features of both non-challenged and challenged broiler jejunum revealed significant improvement along with increased Cu2O and potentiated ZnO doses. Supplementation of Cu2O and potentiated ZnO decreased the jejunum structure damages and intestinal lesion score (P < 0.002). Eimeria caused a decrease (P < 0.006) in total antioxidant capacity. Superoxide dismutase increased by dietary zinc supplementation (P < 0.05). Results suggested that a combination of Cu2O and potentiated ZnO could exhibit efficient anticoccidial activity.

The effects of glutathione supplementation on post-thawed Turkey semen quality and oxidative stress parameters and fertilization, and hatching potential.

This study was conducted to investigate the effect of semen extenders enriched with glutathione (GSH) on in vitro quality parameters and fertility of post-thawed turkey. In experiment 1, pools of semen diluted in glucose-based extender containing 0.5, 1, and 2 mM of GSH were cryopreserved. During the next step, a different variable such as motility and motion parameters, plasma membrane integrity (PMI) and functionality (PMF), DNA integrity, lipid peroxidation (MDA), total antioxidant capacity (TAC), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity were determined in the post-thawed samples. In the second experiment, artificial insemination was used to evaluate the fertility and hatchability performances of the post-thawed semen. The results of the first experiment showed that the extenders supplemented with 2, 1 and 0.5 mM of GSH had higher levels (p ≤ 0.05) of motility and motion parameters, PMI, PMF, TAC, CAT and SOD activity and lower abnormal morphology, DNA damage, and lipid peroxidation respectively in comparison to the control group (only extender with semen). Notably, the second experiment showed a higher rate of fertility (p ≤ 0.05) in 2 mM of GSH compared to the control group. It can be concluded that adding 2, 1 and 0.5 mM of glutathione leads to an improvement in the survival of the post-thawed turkey, while 2 mM of GSH can increase the fertility strength of the turkey sperm; hence it can be used to improve fertility and hatchability performance.

Supplementation of Plant-Based Cryopreservation Medium with Folic Acid Conserves the Quality of Bulk Post-Thawed Spermatozoa.

The current study evaluated the effects of cryopreservation medium supplementation with folic acid as an antioxidant on post-thawed semen quality in bulk. Semen samples were collected from four proved Iranian Mahabadi bulls and diluted in extender containing 1.5% soybean lecithin. The diluted semen was assigned into six parts and supplemented with different doses of folic acid as follows: FA0 (extender without folic acid), FA0.05, FA0.1, FA0.2, FA0.4, and FA0.8 (extenders containing 0.05, 0.1, 0.2, 0.4, and 0.8 mM folic acid, respectively). Then, the semen samples were cryopreserved in liquid nitrogen. Sperm motility and velocity parameters, membrane integrity, abnormal morphology, viability, and lipid peroxidation were evaluated after thawing. In the results, FA0.05 presented higher (p≤0.05) total motility, progressive motility, membrane integrity, and viability and lower lipid peroxidation compared to other groups. Abnormal morphology was not affected (p>0.05) by treatments. In conclusion, supplementation of cryopreservation medium with 0.05 mM folic acid is a helpful method to conserve the quality of post-thawed semen in bulk.

Effects in broiler hens of genetic lines differing in fertility, biotin supplementation, and age on relative abundance of oviductal transforming growth factor-ß and carbonic anhydrase mRNA transcripts.

There was evaluation of effects of biotin administration on oviductal abundance of transforming growth factor-ß (TGF-ß) and carbonic anhydrase (CA) mRNA transcript in younger and older broiler hens of relatively lesser and greater fertility lines. Additionally, effects of biotin supplementation on attenuation of age-related subfertility were evaluated. Hens from the relatively greater (Line D, n = 60) and lesser (Line B, n = 60) fertility rate line were randomly assigned to three treatment groups. Biotin was not or was administered in drinking water from 30 to 33 (younger age) and 53 to 56 (older age) wk of age to have access to no biotin (T0), or 0.3 (T1), or 0.45 (T2) mg/L of biotin. There was assessment the relative oviductal abundances of TGF-ß and CA mRNA transcript abundances. Supplemental biotin and age had no effect on the relative abundance of oviductal TGF-ß mRNA transcript in hens of Line D. There, however, was a ten-fold greater abundance of TGF-ß in hens of the T0 group of Line B compared with Line D. Relative abundance of TGF-ß mRNA transcript was greater in younger hens of Line B; however, biotin supplementation of older hens of the T2 group of Line B resulted in a similar TGF-ß abundance to that of younger hens. Inconstant with the TGF-ß abundance, CA abundance in hens of Line B was not affected by supplemental biotin or bird age. Overall, differences in TGF-ß or CA abundances did not affect fertility of broiler hens.

The effect of dietary coenzyme Q10 on plasma metabolites and hepatic gene expression in broiler breeder hens.

1. This study was performed to evaluate the effects of dietary supplementation of coenzyme Q10 (CoQ10) on laying rate, body weight, plasma metabolites and some liver gene expression in broiler breeder hens. 2. A total of 128 broiler breeder hens (Arbor Acres Plus, 47 weeks of age) were randomly distributed to four dietary groups supplemented with different levels of CoQ10 (0, 300, 600 or 900 mg/kg diet) with four replicates of eight hens each. During 47-54 weeks of age, laying rate, egg mass and body weight were recorded weekly. To assay plasma biochemical indicators, blood samples were collected at 54 weeks of age. At the end of the experiment, for evaluating the abdominal fat weight, liver weight and expression of the adiponectin and proliferator-activated receptor-α (PPAR-α) genes in the liver, eight hens per treatment were selected, weighed and humanely killed by decapitation. 3. Dietary supplementation of CoQ10 linearly decreased abdominal fat weight, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities by increased levels of CoQ10. The plasma levels of glucose, cholesterol and alkaline phosphatase (ALP) activity were quadratically decreased by increased levels of CoQ10. The best plasma levels of glucose, cholesterol and ALP activity were estimated at 562.5, 633.3 and 517.8 mg CoQ10/kg diet, respectively. Adiponectin and PPARα gene expression exhibited a linear increased by increased levels of CoQ10. 4. In conclusion, addition of CoQ10 to the diet influenced lipid metabolism and expression of the adiponectin and PPAR-α genes, which might be partially due to the improvement in mitochondrial metabolism and energy production. However, further studies are necessary to determine the effects of CoQ10 on these indicators in broiler breeder hens during ageing.

The effect of zinc oxide on rooster semen cryopreservation.

1. Deleterious effects from the freeze-thawing process on post-thawed sperm quality attributes are main limiting factors in cryopreservation. The current study was conducted to determine the effect of semen extender containing zinc oxide (ZnO) on post-thaw rooster sperm quality indices.2. Semen samples from six, 60-week-old broiler breeder roosters were collected weekly during five successive weeks. The samples were mixed and divided into three equal parts and diluted with semen extender containing different levels of ZnO; 0 (ZnO-0), 1 (ZnO-1) or 2 (ZnO-2) µg/ml. After thawing, motility and velocity parameters, plasma membrane functionality, apoptotic like changes, mitochondrial membrane potential (MMP), and DNA fragmentation index (DFI) were evaluated.3. Results showed that the addition of ZnO in the extender quadratically affected (P < 0.01) total motility (TM), progressive motility (PM), and average path velocity (VAP) with the highest values were noted in the ZnO-1 group. Levels of ZnO quadratically affected percentages of live (P < 0.01), apoptotic (P < 0.03) and dead (P < 0.10) spermatozoa, where the highest percentage of live, and the lowest percentage of apoptotic or dead spermatozoa was for the ZnO-1 group. Although adding ZnO quadratically affected plasma membrane functionality and MMP (P < 0.01), it did not affect (P > 0.05) DFI.4. In conclusion, there were some beneficial effects of ZnO supplementation in semen extender on post-thawed rooster sperm quality which may result in a better freezability.

Changes in broiler breeder hen's immunity by zinc oxide and phytase.

BACKGROUND: The immune response of aged broiler breeder hens is influenced by many factors including obesity and aged lymphatic organs, but may improve by increasing the bioavailability of various nutrients such as zinc (Zn). Dietary supplementation of phytase can improve Zn availability in senescent broiler breeder hens. AIMS: The aim of this study was to investigate the effect of supplementary zinc oxide (ZnO) and phytase in a maize-soybean meal-based diet on immune responses of broiler breeder hens. METHODS: In a 2 × 4 factorial arrangement, a total of 128 hens were randomly assigned into eight groups. The birds received two levels of phytase (0 or 300 U/kg diet) and four levels of ZnO (30, 60, 90, and 120 mg/kg diet) for 13 successive wk (59-72 wk of age). RESULTS: Results showed that phytase supplementation significantly increased immunoglobulin M (IgM), cutaneous basophil hypersensitivity (CBH) responses, total number of leukocytes, percentage of lymphocytes, and heterophil to lymphocyte ratios. The percentage of basophils and monocytes, however, decreased with phytase supplementation. Supplementation of ZnO increased anti-sheep red blood cells (SRBC) antibody titer, IgM, CBH responses, the total number of leukocytes, and the percentage of lymphocytes. Dietary supplementation of ZnO decreased the percentage of heterophil, and heterophil to lymphocyte ratio. A significant interaction effect of phytase and ZnO was found on the total number of leukocytes and percentage of lymphocytes. CONCLUSION: Dietary supplementation of ZnO (90 mg/kg diet) and phytase had some positive effects on improving immune responses in broiler breeder hens.

Effects of live yeast dietary supplementation on hormonal profile, ovarian follicular dynamics, and reproductive performance in dairy cows exposed to high ambient temperature.

Two experiments were conducted to investigate the effects of dietary supplementation with live yeast culture on hormonal profile, ovarian follicular dynamics and reproductive performance of lactating dairy cows exposed to high ambient temperature. In Expt. 1, Holstein cows were fed a diet with (n = 6) or without (n = 6) 4 g yeast/h/d from 21 d before expected calving date until 8 weeks postpartum. Concentrations of glucose, insulin, insulin like growth factor-I (IGF-I), estradiol-17ß (E-17ß) and progesterone (P4) were determined in plasma, and transrectal ultrasonography was used to monitor ovarian follicular dynamics. In Expt. 2, 150 Holstein cows (75/dietary group) received same diets as Expt. 1 with the exception that diets were fed from d 1 to d 70 postpartum. The effect of dietary treatments on reproductive performance was examined. In Expt. 1, plasma IGF concentrations during dietary treatment were higher (P = 0.05) in cows fed diets supplemented with yeast. On d 60 postpartum, average plasma concentrations of glucose (48.3 vs. 41.0 mg/dL) and insulin (0.90 vs. 0.23 µU/mL), were also greater in cows fed diets supplemented with yeast compared to that in cow fed diets without supplementation. Similarly, plasma concentrations of E-17ß at estrus (P = 0.016) and P4 on d 10 of the estrous cycle (P = 0.021) were greater in yeast-supplemented cows. Moreover, yeast supplementation resulted on average in an estrous cycle 2.6 d shorter (P = 0.05) and in the development of larger ovulatory follicles (18.4 vs. 17.2 mm in diameter; P < 0.01). In Expt. 2, days open were fewer and the proportion of pregnant cows at 120 and 150 d postpartum were greater (P < 0.01) in cows fed diets supplemented with yeast compared to those receiving diets without yeast. In summary, cows fed diets supplemented with yeast had greater plasma IGF-I, E-17ß and P4 concentrations, larger ovulatory follicles, shorter estrous cycles, and improved reproductive performance. Therefore, live yeast dietary supplementation could enhance lactating dairy cow's fertility during heat stress through improvement in hormonal profile and development of larger ovulatory follicles.

Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin- and egg yolk-based extenders.

Ram semen cryopreservation is not efficient for artificial insemination in commercial herds. Beneficial effects of dietary fish oil have been evaluated for cryopreservation of ram semen in soybean lecithin (SL) and egg yolk (EY)-based extenders. A factorial study (two diets × two extenders) was used to analyze the effects of two diets supplemented with fish oil (n-3 fatty acid) or palm oil (saturated fatty acids; [SFAs]) to freeze ram semen in two extenders containing SL or EY. Motility characteristics, membrane integrity, abnormal morphology, mitochondria activity, acrosome integrity, apoptotic status, and fertilizing ability were assessed after freeze-thawing. Although diet had significant (P ≤ 0.05) effects on the quality parameters of frozen-thawed sperm, effects of extenders on these traits were not significant (P > 0.05). The higher significant (P ≤ 0.05) percentage of total motility and progressive motility were observed in n-3/SL (44.83 ± 1.56 and 28.33 ± 1.4) and n-3/EY (43.33 ± 1.56 and 28.50 ± 1.4) than SFA/SL (32.16 ± 1.56 and 14.00 ± 1.4) and SFA/EY (31.66 ± 1.56 and 12.66 ± 1.4) groups. Moreover, n-3/SL and n-3/EY produced the higher significant (P ≤ 0.05) percentage of membrane integrity of sperm (39.83 ± 1.4 and 37.33 ± 1.4) than SFA/SL and SFA/EY (29.83 ± 1.4 and 28.5 ± 1.4). For viability results, the higher significant percentage of live sperm was observed in n-3/SL and n-3/EY (43.16 ± 1.38 and 45.66 ± 1.38) than SFA/SL and SFA/EY (28.66 ± 1.38 and 27.5 ± 1.38). For fertility trials, n-3-based diets (n-3/SL and n-3/EY) improved significantly (P ≤ 0.05) pregnancy rate (44% and 46%), parturition rate (42% and 42%), and lambing rate (46% and 44%) compared with the SFA-based diets (SFA/SL and SFA/EY). No interaction effects have been found between diets and extenders (P > 0.05). It seems that dietary fish oil can improve the semen performance after freezing-thawing process and artificial insemination aside from type of extenders.

Effect of feeding guanidinoacetic acid and L-arginine on the fertility rate and sperm penetration in the perivitelline layer of aged broiler breeder hens.

Two experiments were conducted to evaluate the effects of feeding guanidinoacetic acid (GAA) and L-arginine (ARG) on fertility and sperm penetration (SP) rate of broiler breeder hens. In the first experiment, a total of 200 broiler breeder hens (Ross 308) aged 53 weeks were randomly allotted to four dietary treatments (0, 0.6, 1.2 and 1.8 g GAA/kg diet) with five replicates of 10 birds each. In the second experiment, 320 broiler breeder hens (Ross 308) were used from 53 to 62 weeks of age in a 2 × 4 factorial arrangement (0 or 1.2 g GAA/kg diet along with 0, 3, 6 or 9 g ARG/kg diet). The hens received a diet containing 2800 kcal ME/kg and 14% CP. Sixteen sexually mature Ross 308 breeder roosters (34 weeks old) were used to artificially inseminate the hens. Fertility of the hens was determined in 61 and 62 weeks of age. The sperm penetration holes in the inner perivitelline layer (IPL) overlying the germinal disc were enumerated on days 3 and 7 following each insemination. Adding GAA to the breeder diet increased the number of SPs in the IPL and fertility in both experiments (p < 0.01). The interactive effect of ARG and GAA on the SP and fertility was significant. Supplementary ARG increased the SP rate in the IPL (p < 0.01). In conclusion, dietary supplementation of GAA and ARG might be potentially used to improve the fertility of broiler breeder hens at the later phase of the egg production period.

Improvements in semen quality, sperm fatty acids, and reproductive performance in aged Cobb 500 breeder roosters fed diets containing dried ginger rhizomes (Zingiber officinale).

Exposure to high levels of polyunsaturated fatty acid predisposes spermatozoa to lipid peroxidation, resulting in their decreased fertility. Ginger powder (GP), which is high in antioxidative compounds, was fed to aged breeder roosters to improve their reproductive performance. Seventy-five 52-wk-old Cobb 500 breeder roosters randomly received either 0 (GP0), 15 (GP15), or 30 (GP30) g of GP/kg of diet for 14 consecutive wk, during which time their seminal characteristics were evaluated every 2 wk. At the end of the trial, semen samples were tested for determination of sperm fatty acid (FA) concentration and seminal plasma total antioxidant capacity. Furthermore, sperm penetration was assayed, and using 225 artificially inseminated hens, fertility and hatchability rates were determined. Dietary GP improved sperm forward motility, live sperm percentage, and sperm plasma membrane integrity. These were associated with a decrease in the percentage of abnormal sperm. The seminal TBA reactive species concentration was lower in birds belonging to the GP30 treatment in comparison with those in the GP15 and GP0 treatments. The feeding of GP resulted in overall decreases and increases in sperm saturated and unsaturated FA, respectively. The n-6:n-3 FA ratio of sperm was decreased in the GP30 group in comparison with controls. The highest levels of sperm C20:4(n-6) and C22:6(n-3) FA were recorded in the GP15 and GP30 treatments, respectively. A higher percentage of sperm C22:4(n-6) FA was found in GP-fed roosters. Seminal plasma total antioxidant capacity was considerably improved by the GP15 and GP30 treatments. Further, a higher number of perivitelline membrane sperm penetration holes was recorded for the GP30 treatment in comparison with the GP15 and GP0 treatments. Interestingly, although hatchability, chick quality, and embryonic mortality were not affected by dietary treatment, fertility rate was improved by the feeding of GP. In conclusion, dietary GP improved most of the seminal characteristics evaluated in aged roosters of this study, suggesting that it has potential for use in attenuating age-related subfertility in senescent male commercial broiler breeders.