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.

Combined effect of probiotics and specific immunoglobulin Y directed against Escherichia coli on growth performance, diarrhea incidence, and immune system in calves.

Enterotoxigenic Escherichia coli (ETEC) K99 is one of the major pathogens associated with calf diarrhea. The induction of passive immunity in animals by immunoglobulin Y and using probiotics are inexpensive alternatives to antibiotics for the prevention and treatment of a number of bacterial infections, including diarrhea. Hence, the aim of this research was to evaluate the impact of dietary probiotics and ETEC K99-specific egg yolk antibody supplements, alone and in combination with each other, on health and growth parameters, diarrhea incidence and immune stimulation in newborn Holstein calves. One hundred and twenty neonatal calves were allocated randomly into 4 dietary groups (n = 30 per group) received colostrum/milk without any additives (control group), or supplemented with egg yolk powder contained E. coli K99-specific antibody (Ab group; 1 g/day), a commercial probiotic, Hypro-calves (Pro group; 3 g/day), and their combination (Ab+Pro group), from day (d) 1 to d28 of age. Analyses of the growth parameters, feed efficiency, fecal score, and microbiota and immune function were carried out on d0, 14, 21, and 28 of the experiment. Calves in Ab or Ab+Pro group had higher (P < 0.05) average daily gain compared to control and Pro groups during 0-14d. Feed efficiency of calves in Ab and Ab+Pro groups was significantly higher than that in control group during the period of 0-14d; however, no significant differences were observed in 0-28d period. Diarrhea prevalence and fecal score in Ab+Pro group were lower than control group (P < 0.05). Calves in Ab+Pro group had the lowest number of fecal E. coli in comparison to other groups on d28 (P < 0.05). Feeding Ab+Pro supplement increased (P < 0.05) concentrations of blood IgA and serum CD4 compared to the control group. Likewise, calves in Pro group had higher CD4 levels as compared to the control calves (P < 0.05). Serum concentration of interferon-gamma in control group was lower than other groups (P < 0.05). Overall, these data suggest that feeding a combination of probiotic and specific antibody against ETEC to neonate Holstein calves enhances feed efficiency, boosts immunity, and reduces diarrhea prevalence.

Improvement of in vitro proliferation of cockerel spermatogonial stem cells using different combinations of growth factors.

1. This study examined whether in vitro proliferation and maintenance of cockerel spermatogonial stem cells (SSCs) could be improved by adding different combinations of growth factors (GFs), including glial cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF) or leukaemia inhibitory factor (LIF) into the culture medium. 2. The SSCs were isolated from the testes of immature cockerels. For short-term cultures, a medium supplemented with different combinations of GFs for 7 d in 5 replicates was used. The groups were classified as follows: without GF (control group); with GDNF (G group); with GDNF and bFGF (GF group); and with GDNF, bFGF and LIF (GFL group). The number of colonies and cells per colony, as well as the transcript abundance of STRA8 and OCT4 genes, was determined 7 d after the initial culturing. Immunofluorescence staining of SSEA-1, SSEA-3 and VASA protein markers, besides periodic acid-Schiff (PAS) staining, was carried out. 3. The number of colonies and cells per colony increased in the G, GF and GFL groups, compared to the control group (P < 0.01); however, the highest proliferation and colony formation were observed in the GFL group. The positive immunofluorescence staining of SSEA-1, SSEA-3 and VASA protein markers, as well as PAS staining, confirmed the self-renewal and colonisation of cockerel SSCs. The proliferation results were supported by the increased STRA8 and OCT4 transcript abundance in the treated groups (G, GF and GLF), compared to the control group. The SSC proliferation was associated with the higher transcript abundance of STAR8 and OCT4 genes in the GFL group, compared to the G and GF groups (P < 0.01). 4. The results showed that proliferation and colony-forming capacity of cockerel SSCs were positively improved by GDNF, bFGF and LIF. However, the most significant effect was observed when the medium was supplemented with LIF in combination with GDNF and bFGF.

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.

Effect of Glutathione Supplementation to Semen Extender on Post-thawed Rooster Sperm Quality Indices Frozen After Different Equilibration Times.

BACKGROUND: Avian sperm is susceptible to lipid peroxidation, compromising their fertility. The semen antioxidant system protects sperm plasma membrane against reactive oxygen species. OBJECTIVE: The study evaluates the effect of glutathione (GSH) addition to semen extender during different equilibration times (ET) on rooster sperm cryopreservation. MATERIALS AND METHODS: Semen samples are weekly collected from 60-week-old broiler breeder roosters. Collected samples were pooled and divided to six equal parts and frozen according to a randomized design (2 × 3 factorial arrangement). Treatments included adding two levels of GSH [0 (GSH-0) or 1 (GSH-1) mM] to semen extender during three ET: 0 (ET-0), 4 (ET-4) or 8 (ET-8) hours. Post-thawed motility and velocity parameters, apoptotic like changes, plasma membrane functionality, and mitochondrial membrane potential (MMP) were evaluated. RESULTS: Post-thawed total motility is improved in the GSH-1 compared to the GSH-0 group (P<0.10). Total motility responded quadratically to increasing levels of ET such that the highest value is recorded at ET-0. Although progressive motility (PM) is not affected by GSH or ET, the highest PM is obtained in the GSH-1×ET-0 group (P<0.05). The VAP and STR is improved in the GSH-1 compared to GSH-0 group; however, VAP decreases quadratically, and STR decreases linearly as ET is advanced (P<0.05). The interactive effect of GSH by ET tends (P<0.08) to affect the wobble coefficient (WOB), such that the highest value is recorded in the GSH-1×ET-0 group. Within both GSH supplemented and control groups, the amplitude of lateral head displacement (ALH) is highest (P<0.05) in the ET-0 group. The percentage of live spermatozoa quadratically decreases and the percentage of dead sperm quadratically increases in response to graded levels of ET (P<0.01). The highest plasma membrane functionality is also noted in the GSH-1×ET-0 group (P<0.05). Mitochondrial membrane potential quadratically decreases in response to increasing levels of ET (P<0.05). CONCLUSION: Generally, GSH supplementation to rooster sperm extender has some beneficial effects on post-thawed sperm motion characteristics, but does not positively interact with ET.

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.

The long-term oral administration of thyroxine: effects on blood hematological and biochemical features in broiler breeder hens.

Published data on the beneficial effect of short-term administration of thyroxine (T4) in broiler breeder hens to reduce the ascites incidence in their progeny chicks raises the question as to what extent might the long-term maternal administration of T4 affect the blood hematological and biochemical attributes in breeder hens. A total of 70 broiler breeder hens (47-wk-old) were randomly allotted to control or thyroxine treated (T4) groups. Pure T4 (0.3 mg/bird per day) was orally administered to T4 birds for 14 successive weeks, whereas the control group received the drinking water only. Blood samples were obtained from the brachial vein prior to the initiation of the trial as well as weeks 50, 53, 55, 57, 59, and 61 of age. Body weight was decreased but egg production was not affected by T4 treatment. Plasma concentration of T4, but not triiodothyronine (T3), was increased in T4-treated hens (P < 0.05). The total number of leukocytes and erythrocytes were also higher in T4 birds. A significant effect of time was observed for erythrocyte number and plasma cholesterol concentration (P < 0.05). The long-term administration of T4 did not affect the concentrations of serum calcium and plasma total protein, albumin, globulin, cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, very low density lipoprotein, alanine amino transferase, and aspartate amino transferase (P > 0.05). However, serum concentrations of phosphorus, glucose, and alkaline phosphatase were higher in T4 hens as compared to their control counterparts. In spite of differences in circulatory concentrations of a number of traits between the experimental groups, the recorded values were within their reference ranges. Therefore, the administration of T4 for an extended period of time had no apparent adverse effect on the clinical profile in subjected hens, which may practically support the implementation of this preventative treatment as an approach to decrease the ascites incidence; however, a lower incidence rate in the progeny chicks produced from hens receiving T4 for long-term periods of time remains to be elucidated.

TGF-ß4 and HSP70 responses in breeder hens treated with thyroxine.

A hypothesis was tested that long-term administration of thyroxine (T4) in broiler breeder hens would affect fertility, sperm penetration rate, and the duration of fertility. Relative abundance of oviductal TGF-ß4 and HSP70 mRNA was determined to ascertain whether T4 treatment affected these genes, and modulated the sustained storage of spermatozoa within the uterovaginal sperm storage tubules of hens. A total of 70, 47-week-old Cobb 500 breeder hens was randomly allotted to two treatment groups (T4 treatment (ET) and control). The T4 was orally administered to the ET group (0.3 mg T4/bird/day) for 100 consecutive days; whereas the control group was not administered T4 during the experimental period. Breeder hens were artificially inseminated to evaluate specific reproductive variables. On the last day of the treatment period two hens /replicate were randomly killed to estimate oviductal gene expression. The T4 treatment resulted in an increase in plasma concentration of T4; however, the T3 concentration was not affected. The long term administration of T4 had no effect on fertility; however, it resulted in a decreased sperm penetration rate and decreased the duration of fertility compared with the control group. The relative abundance of TGF-ß4 and HSP70 mRNA in the SST was not influenced by T4 supplementation. The correlation coefficients between fertility and sperm penetration rate with relative abundance of TGF-ß4 and HSP70 were not significant. Overall, among the diverse reproductive variable assessed in the current study, the sperm penetration rate and the duration of fertility were most responsive to long-term treatment with T4.

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.

Histomorphometrical changes of oviduct during the long-term exposure of breeder hens to extra thyroxine.

In this research a 100 day long treatment period was considered to unmask the probable adverse effects of long-term induced hyperthyroidism on histomorphometrical attributes of the oviduct in broiler breeder hens. A total of seventy 47-week-old Cobb 500 breeder hens were randomly allotted to two treatment groups (5 replicates of 7 hens each). Thyroxine (T4) was orally administered to the hyperthyroid group (0.3 mg/bird/d) for 100 consecutive days; whereas the control group received drinking water only throughout the trial. At 64 weeks of age, 2 birds per replicate were killed by cervical dislocation and their oviducts were removed. For histomorphometrical observations, segments were taken from five different regions. After tissue preparation and staining with haematoxylin and eosin, histological layers were evaluated using light microscopy. The assessment of histomorphometrical characteristics of oviduct showed the height of mucosal folds in the magnum, thickness of mucosal folds of the magnum and uterus, thickness of tunica muscularis in the magnum and vagina, epithelial thickness of the isthmus and vagina, and uterine tubular glands percentage were decreased in the hyperthyroid birds compared with the control counterparts. The results showed long-term induced hyperthyroidism was associated with a decrease in a number of histomorphometrical traits in different regions of the oviduct. Some studies should be done to clarify to what extent the long-term maternal hyperthyroidism might affect the egg production, fertility rate, duration of fertility, and sperm penetration rate to make a final decision on exploitation of this preventative treatment to diminish the ascites incidence in progeny chicks.

Orally administered Chrysin improves post-thawed sperm quality and fertility of rooster.

Chrysin is a bioflavonoid compound found in passion flower, chamomile, propolis and honey at high levels. Post-thawed sperm quality and fertility of Chrysin-fed roosters were assessed in this study. Twenty 40-week-old male broiler breeders were randomly divided into four groups and fed basal diet supplemented with different levels of Chrysin including 0 (Ch-0), 25 (Ch-25), 50 (Ch-50) or 75 (Ch-75) mg/day for 12 consecutive weeks. Semen samples were weekly collected from 6th to 9th week of experiment to evaluate some sperm quality parameters including total and progressive motility, plasma membrane integrity and functionality (in fresh and post-thawed samples) and mitochondrial activity (only in post-thawed samples). Also, collected semen samples from 10th, 11th and 12th week of experiment were frozen and then artificially inseminated to test fertility rate. According to the results, an improvement in both fresh and post-thawed sperm quality including total [fresh: 88.00 ± 0.58 and 87.25 ± 0.67 (p < .01); post-thawed: 51.07 ± 2.05 and 52.72 ± 1.96 (p < .01)] and progressive motility [fresh: 76.00 ± 0.58 and 78.25 ± 0.65 (p < .01); post-thawed: 40.61 ± 2.01 and 39.88 ± 2.01 (p < .01)], plasma membrane integrity [fresh: 91.60 ± 0.58 and 89.85 ± 0.59 (p < .01); post-thawed: 56.99 ± 1.86 and 54.39 ± 1.86 (p < .01)] and functionality [fresh: 75.40 ± 0.42 and 77.90 ± 0.96 (p < .01); post-thawed: 45.69 ± 1.71 and 46.35 ± 1.71 (p < .01)] was noted for both Ch-50 and Ch-75, respectively, groups compared to control group. Despite no significant change in mitochondrial activity, fertility rate of post-thawed spermatozoa was significantly improved in all Chrysin-fed groups compared to Ch-0 group. In conclusion, oral Chrysin administration to roosters could ameliorate cryopreservation-induced impairment of sperm quality and fertility rate.

Effect of tert-butyl hydroquinone on bull semen cryopreservation.

BACKGROUND: Many studies have been shown that freezing induced oxidative stress has detrimental effect on post-thaw sperm quality. OBJECTIVE: This study was conducted to investigate the effect of tert-butyl hydroquinone (tBHQ) on bull semen crtopreservation. MATERIALS AND METHODS: In this study, four different levels of tBHQ [Optidyl containing zero (T0), 2.5 (T2.5), 5 (T5), and 7.5 µM (T7.5) tBHQ] was used to study the effect of tBHQ on freezability of bull semen. On each collection day, four ejaculates were collected (a total of 24 ejaculates from four bulls), pooled and divided to four equal parts. Each part was diluted with one of the above-mentioned extenders and frozen. After thawing, sperm motility, plasma membrane functionality and integrity, apoptosis status and mitochondrial activity were assessed. RESULTS: The results show that total sperm motility was significantly higher in T5 compared to other groups. The value of VSL was significantly lower in T5 compared to T0. Also, T5 resulted in lower LIN and STR versus T0 and T2.5 groups. All extenders containing tBHQ resulted in a significantly higher percentage of sperm with functional membrane compared to T0 groups. Finally, Apoptosis related parameters and mitochondrial activity were not significantly difference between the groups. CONCLUSION: adding 5 µM tBHQ to the bull semen extender can be beneficial for post-thaw sperm quality. Also, in vivo or in vitro fertility test is recommended to test fertilizing ability of tBHQ exposed sperm.

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.

Protective role of glutathione in buck semen cryopreservation.

The aim of this study was to determine the effect of low levels of glutathione on post-thawed buck sperm quality. In this experiment, different concentrations of glutathione [0 (LG-0), 0.5 (LG-0.5), 1 (LG-1), 1.5 (LG-1.5), and 2 (LG-2) mM] were added in a soybean lecithin-based extender. A total of 16 ejaculates from four bucks were collected and pooled. Each pooled sample was divided into five equal parts and each part was diluted by one of the above mentioned groups. After freeze-thawing process, motility and velocity, plasma membrane integrity and functionality, and apoptosis features of spermatozoa were evaluated. The results of this experiment showed that total motility (50.75 ± 2.33), plasma membrane integrity (55.75 ± 3.01) and functionality (46.75 ± 2.79) were higher in LG-1 extender compared to other extenders (P<0.05). The percentage of live spermatozoa (53.23 ± 3.26) was higher in LG-1 extender compared to other extenders, with the exception of LG-1.5 extender (P<0.05). Also, the percentage of late apoptotic spermatozoa (21.33 ± 1.63) was lower in LG-1 extender compared to other extenders (P<0.05). In conclusion, our results showed that GL-1 extender resulted in higher post-thawed buck sperm quality compared to other extenders.