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BACKGROUND: In ovo injection of nutrients as an early feeding method in birds directly supplies the nu- trients to the developing embryo. OBJECTIVES: This study was designed to evaluate the effects of in ovo injection of nicotonic acid, panto- thenic acid and folic acid on the performance and immune system of broilers. METHODS: 450 Ross 308 fertile eggs were divided into 5 groups and placed in a hatchery machine. Five experimental groups included in ovo injection of 0.121 mg of nicotonic acid, 0.052 mg of pantothenic acid, 0.007 mg of folic acid on the 14th day of incubation period, positive control or injection control (physiological serum injection) and negative control (non injecting control). RESULTS: At the age of 18 days of the rearing period, injection of pantothenic acid and nicotinic acid in- creased the antibody titre against Newcastle Virus and folic acid and pantothenic acid reduced the SRBC titer. At 35 days of age, nicotinic acid and folic acid had lower SRBC titer than the negative control group. The highest lymphocyte to heterophilia ratio was observed in the pantothenic acid group and the lowest levels were seen in the folic acid group. In ovo injection of nicotinic acid and pantothenic acid caused weight loss in chicks during the first and second weeks of rearing period compared to positive and negative controls groups. CONCLUSIONS: The results of this study indicated a positive effect of in ovo injection of pantothenic acid and nichotinic acid on some immune parameters of broiler chicks. In despite of the negative effect of in ovo injection of nicotinic acid and pantothenic acid on growth rate of chicks during the first and second week of age, there was a compensatory growth for the nicotinic acid group such that this treatment positively influenced the final weight of the broilers.

Objective of experiment was to determine whether oxidative stress (OS) and inflammation altered embryonic loss in dairy cows. Blood samples were collected at days 0, 16, 32 and 60 after timed (AI) from 200 Holstein cows to determine embryonic loss based on interferon-stimulated gene-15 (ISG15) mRNA expression (day 16) and ultrasound at day 32 and day 60. Leucocyte expressions of mRNA TLR2, TLR4, TNF-α, IL1B, IL10, STAT3 (inflammation), PTGS2, PTGES (prostaglandin synthesis), and PLA2G4A and ALOX5AP (eicosanoid metabolism) at days 0 and 16 were determined. Plasma redox status for antioxidant enzymatic activities of glutathione peroxidase (GPX), superoxide dismutase (SOD), total antioxidant capacity (TAC), and concentrations of malondialdehyde (MDA) were determined at days 0, 16, 32 and 60. All antioxidant-redox responses were beneficially significant in pregnant cows diagnosed pregnant at day16 and sustained pregnancy to day 60 compared to non-pregnant cows at day16 or pregnant at day16 and lost embryos by days 32 or 60. The leucocyte mRNA expressions of TLR2, TLR4, STAT 3, IL1B, PTGS2, PLA2G4A and ALOX5AP were greater and PTGES was lower at day16 in pregnant cows that lost embryos early (P

There are several findings indicating that the endocannabinoid system (ECS) is an important factor, acting in multiple ways in regulating reproductive function but changes of this system in the bovine endometrium have rarely been investigated; therefore, this study was designed to consider an association between endometrial ECS expression and different stages of the estrous cycles. MRNA expressions of the ECS were investigated during the late luteal phase and estrus using real-time PCR. Following estrous synchronization of sixteen Holstein dairy cows (34±1.3 kg/day of milk production), using two PGF2α injections given 14 days apart, at 30 and 44 days in milk (DIM), blood samples and ultrasonography (US) were performed every other day from the day of second PGF2α injection (44 DIM) until the start of the next estrous cycle (67±2 DIM) to verify CL development and ovulation. Based on blood and US results endometrial tissue was collected on days 16 (late luteal phase) and 21 (estrus) of the synchronized estrous cycle (ovulation = d 0). Real-time PCR analysis of ECS mRNA expression revealed endocannabinoid receptor (CNR2), diacylglycerol lipase (DAGL), cyclooxygenase-2 (COX-2), fatty acid amide hydrolase (FAAH) and monoglyceride lipase (MGLL) had significant fold differences when comparing two different stages of the estrous cycle (late luteal phase vs. estrus). CNR2 and DAGL showed 2.01 and 2.57 fold increase, respectively (P=0.04 and P=0.02), in estrous cows. Among the analyzed genes FAAH (P=0.01) and MGLL (P=0.02) were significantly down-regulated in estrous cows, with a 5.01- and 2.44-fold difference in mRNA expression, respectively. Overall, this study highlights an association between the expression of the ECS in the bovine endometrium and stage of the estrous cycle.

Sustainable production and the increasing number of embryonated hatching eggs are critical aspects of the poultry production industry. The present paper aims to appraise the effectiveness of royal jelly (RJ) on the semen characteristics of Native Mazandaran roosters in both liquid and frozen storage conditions. Semen collected from 10 sexually mature roosters and following dilution was supplemented with RJ at 0.0 (control), 5 (RJ5), 10 (RJ10), 20 (RJ20) and 40 (RJ 40) mg/ml. After cooling and freezing-thawing, the percentage of forward progressive motility, viability, abnormality, hypo-osmotic swelling test (HOST) and the mRNA abundance of antioxidant enzymes of spermatozoa were measured. Our results revealed that the addition of 5 mg/ml RJ to the semen extender significantly increased (p

The aim of the present study was to investigate the effect of dietary supplementation of royal jelly (RJ) on liquid and frozen storage of rooster spermatozoa. Twenty-five 30-week-old of Mazandaran native breeder roosters were randomly divided into five treatments (n = 5 roosters/group). Experimental treatments are designed to include a control group and various levels (0.0 (RJ0), 100 (RJ100), 200 (RJ200), 300 (RJ300) mg kg-1 BW-1 ) of royal jelly (RJ) that were fed to the roosters using force-feed method. The percentage of forward progressive motility, abnormal spermatozoa, membrane integrity and viability of spermatozoa evaluated after 24 and 48 hr of cooling (at 4°C) and after the freeze-thawing process. Also, mitochondrial activity and DNA fragmentation in fresh (24 hr) and post-thawed spermatozoa were assessed. The result of this study showed that the spermatozoa forward progressive motility, abnormality, membrane integrity, and viability were improved by the RJ100 group compared to the other groups after 24 and 48 hr storage period at 4°C. The percentage of membrane integrity and forward progressive motility after freeze-thawing in the RJ100 group was significantly higher than the other groups, and the percentage of abnormal spermatozoa was lower. A significant decrease in semen quality parameters was seen after 24 and 48 hr of refrigeration, but there was no observed change between 2 and 24 hr in the RJ100. The viability percentage of spermatozoa in both RJ100 and RJ200 groups was not different. Moreover, after freeze-thawing, DNA integrity and mitochondrial activity in the RJ100 group were significantly higher than the other groups. According to our results, feeding of RJ at 100 mg kg-1 BW-1 to the roosters was improved spermatozoa characteristics during liquid and cryopreservation conditions.

This study was undertaken to determine the effect of inflammatory status on different patterns of postpartum luteal activity and reproductive performance of Holstein cows during early lactation. The cows (n = 75) averaged 3.4 ± 1.2 (mean ± SEM) in parity and 3.1 ± 0.2 (mean ± SEM) in body condition score at calving. Transrectal ultrasonography was performed twice weekly from day 10-60 postpartum to consider ovarian dynamics. Plasma concentrations of progesterone (P4) and estradiol (E2) were measured twice weekly and plasma levels of tumor necrosis factor alpha (TNF-α) and lipopolysacchride binding protein (LBP) were measured weekly (week 3-6 postpartum). Based on plasma P4 and E2 results, 34 (45.3%) cows had normal luteal activity (NLA), whereas 19 (25.3%), 12 (16.0%), 7 (9.3%) and 3 (4.0%) cows had prolonged luteal phase (PLP), delayed first ovulation (DO), anovulation (AO) and short luteal phase (SLP), respectively. Plasma TNF-α and LBP concentrations were affected by postpartum luteal activity (NLA, SLP and PLP), ovulatory status (DO and AO) and number of weeks postpartum (P .05). These concentrations were greater in cows with PLP (P .05) as compared to NLA cows, and in cows that had delayed ovulation or anovulation compared to ovulated cows (P .05). Cows with PLP had greater open days and lower conception rate as compared to NLA cows (P .05). Healthy cows had a larger CL and greater plasma estradiol and progesterone concentrations at first and second cycle postpartum compared to inflamed cows and followed it with greater fertility (P .05). In conclusion, inflammatory statuses were different in high-producing dairy cows showing PLP, AO and DO in comparison with the postpartum normal luteal activity (NLA) cows that influenced reproduction outcomes.

Royal jelly (RJ) was supplemented to goat oocyte in vitro maturation (IVM) medium at three different concentrations (2.5, 5, and 10 mg/ml). Maturation rate, embryo cleavage, and blastocyst rate were recorded. Gene expression of apoptosis-related transcripts was investigated in matured oocytes. Percentage of oocytes that reached MII-stage was increased in RJ-treated groups compared to the control group. Glutathione (GSH) content of mature oocytes was enhanced when RJ was added to IVM medium at any supplementation compared with control. Percentage of cleaved embryos and blastocysts was higher in the RJ-treated groups at a concentration of 5 than in the 2.5 mg/ml and control group. Total number of cells per blastocyst was not different in the control and RJ-treated group at 5 mg/ml. However, number of apoptotic cells per blastocyst was higher in the control group than in the RJ-treated group at 5 mg/ml. Expression profile of Bax, and p53 was down-regulated while Bcl-2 was up-regulated in oocytes treated with RJ at 5 and 10 mg/ml compared with the control group. Addition of RJ at concentrations of 5 mg/ml improved embryo production through increasing maturation rate. RJ seems to improve the IVM microenvironment by reducing expression of genes inducing apoptosis, enhancing GSH content, and reducing incidence of apoptosis in blastocysts.

This study was designed to evaluate orally administrated Letrozole (Lz) on reproductive performance, plasma testosterone and estradiol concentrations and relative abundance of mRNA of GnRH, FSH and LH in roosters. Ross 308 roosters (n=32) that were 40-weeks of age were individually housed and received a basal standard diet supplemented different amounts of capsulated Lz [0 (Lz-0), 0.5 (Lz-0.5), 1 (Lz-1) or 1.5 (Lz-1.5), mg Lz/bird/day] for 12 weeks. Sperm quality variables and plasma testosterone and estradiol concentrations were assessed from the first to the tenth week of the treatment period. Semen samples from the 11th to 12th week were used for artificial insemination and eggs were collected and allotted to assess fertility and hatchability rates. Relative abundance of hypothalamic and pituitary GnRH, LH and FSH mRNA was evaluated at the end of 12th week. The results indicated that total and forward sperm motility as well as egg hatchability rate were greater in the Lz-0.5 group. Greater sperm concentrations, ejaculate volume, sperm plasma membrane integrity, testis index and fertility rates were recorded for both Lz-0.5 and Lz-1 groups compared with the Lz-0 group (P

The objective was to compare three shortened protocols for timed-AI (TAI) on ovarian responses, pregnancy per AI (P/AI) and pregnancy loss after resynchronization of ovulation in multiparous Holstein cows. Cows (n = 370), at one location, were randomly assigned at non-pregnancy diagnosis (approximately 32 d after AI) to one of three ovulation resynchronization protocols. Cows in the OS group received a 5-d Ovsynch [100 μg GnRH on Day 1, 500 μg cloprostenol (PGF) on Days 6 and 7, GnRH on Day 8.5 and TAI on Day 9 (16 h after second GnRH)]. Cows in the OS + P4 (progesterone) group received a 5-d Ovsynch as described for OS group plus an intravaginal device (Cue-Mate), containing 1.56 g of progesterone (P4), between Days 1 and 6. Cows in the J-synch group received a Cue-Mate and 2.5 mg of estradiol benzoate (EB) on Day 0, PGF and Cue-Mate removal on Day 6, another PGF on Day 7, and 100 μg of GnRH on Day 8.5, with TAI on Day 9. Ovarian response and pregnancy status at 32 and 60 d after the resynchronization TAI were determined by transrectal ultrasonography. Blood samples were collected at first PGF treatment and at TAI from a subset of 40 cows per group to determine P4 concentrations. Percentage of cows with CL at initiation of the protocol did not differ (P > 0.05) among resynchronization groups. Plasma P4 concentrations at first PGF treatment were greater (P < 0.01) in cows that received a Cue-Mate (OS + P4 and J-synch) compared to OS cows. Luteal regression was greater (P < 0.01) for J-synch (88.6%) compared with OS (76.9%) and OS + P4 (78.8%). More (P < 0.01) cows in the OS + P4 and J-synch groups had their estrous cycle synchronized and were pregnant at 32 d after TAI (48.7 and 34.7%, 67.8 and 48.0%, and 72.4 and 50.0% for OS, OS + P4 and J-synch, respectively). However, more cows subjected to J-synch remained pregnant at 60 d after TAI and, hence, had fewer (P < 0.05) pregnancy losses (19.2, 18.8 and 5.0% for OS, OS + P4 and J-synch, respectively). In summary, cows resynchronized with either OS + P4 or J-synch had greater P4 concentrations at first PGF treatment and a greater response to treatments compared to cows subjected to OS. Although OS + P4 and J-synch resynchronization protocols resulted in increased P/AI at 32 d, pregnancy losses were significantly reduced in cows subjected to the J-synch protocol.

The aim of this study was to investigate the effect of different concentrations of royal jelly (RJ) on in vitro maturation (IVM), fertilization, cleavage, blastocyst rates, glutathione (GSH) content in ovine oocyte, mRNA abundance of antioxidant enzymes in both oocyte and cumulus, and glucose metabolism-related genes in cumulus cells. In vitro maturation of oocyte was performed in the presence of control (RJ0), 2.5 (RJ2.5), 5 (RJ5), and 10 (RJ10) mg/mL of RJ. Nuclear status, intracellular GSH content in oocytes, and mRNA abundance of selected genes were evaluated following 24 hours of IVM. Following the IVM, fertilization and embryo culture were carried out in all the groups and embryonic development was examined. The addition of 10-mg/mL RJ to maturation media not only yielded a higher number of oocytes at MII stage but also showed an increased level of intracellular GSH content than did RJ2.5 and control groups. Fertilization, cleavage, and blastocyst rate were higher in the RJ10 treatment group in comparison to the control one. In cumulus cells, the expression of PFKM, PFKL, and G6PDH were increased following the addition of RJ to the maturation media. Supplementation of 10-mg/mL RJ to IVM medium increased the GPx mRNA abundance in both oocyte and cumulus cells and SOD expression in the cumulus cells. The CAT mRNA abundance was not influenced by the addition of RJ to the maturation media in either oocyte or cumulus cells. It seems that the improvement of oocyte maturation and its subsequent development in RJ10 group may be associated with amelioration of redox status in the oocytes and activation of glucose metabolic pathways in their surrounding cumulus cells.

The role of oxidative stress in the control of female reproduction has not been fully elucidated in ruminants; however, it seems that antioxidants can make influence to the reproductive axis at different levels. Therefore, the aim of this study was to determine the relationship between antioxidant status and concentrations of trace minerals (chromium (Cr), copper (Cu), iron (Fe), and zinc (Zn)) with postpartum luteal activity and fertility in Holstein dairy cows. The cows (n = 100, a parity range of 2–5, and a body condition score (BCS) of 3.0 ± 0.25, mean ± SEM) were assigned to the experiment at second week post calving. Ovarian follicular dynamics were monitored twice a week (from 3rd to 6th weeks post calving) by transrectal ultrasonography (US). Blood samples were collected twice weekly from the 3rd to the 6th weeks post calving at timed artificial insemination (TAI), and days 32 and 50 post AI to determine superoxide dismutase (SOD), glutathione peroxidase (GPX), total antioxidant capacity (TAC), and trace mineral concentrations. There were associations between plasma concentrations of SOD, GPX, and TAC with postpartum luteal activity (PLA, P = 0.01) and ovulation (P = 0.03). Mean plasma SOD and GPX activities and TAC levels (U/mL) were greater in cows with normal luteal activity (NLA) than prolonged luteal phase (PLP) and anovulation (AO) cows, as well as in ovulated compared to AO cows (P = 0.03). Pregnant cows had greater levels of SOD, GPX, and TAC (U/mL) at TAI than non-pregnant cows (P = 0.01). Plasma Cu and Zn concentrations increased in pregnant compared to non-pregnant cows at TAI. In conclusion, antioxidant levels and Cu and Zn concentrations were associated with PLA and fertility.

Optimizing culture conditions lead to the improvement of oocyte developmental competence and additives with anti-oxidative activity in culture media improved embryonic development. Royal jelly (RJ) is a product from the cephalic glands of nurse bees that has considerable health effects. The aim of this study was to investigate the effect of different concentrations of RJ on the maturation, cleavage, and blastocyst rates and gene expression in the oocyte and cumulus cells during in vitro maturation (IVM) of sheep oocyte. IVM of oocyte was performed in the presence of control (RJ0), 2.5 (RJ2.5), 5 (RJ5), 10 (RJ10), 20 (RJ20), and 40 (RJ40) mg/mL of RJ. Following the maturation period, parthenogenetic activation was carried out in two treatment groups (RJ0 and RJ10) and embryonic development was examined three and eight days thereafter. Moreover, the relative expression of BCL2 and BAX in oocyte as well as BCL2, BAX, HAS2, PTGS2, and STAR in cumulus cells were assessed. The results indicated that the addition of 10 mg/mL of RJ (90 ± 4.51%) to the maturation medium linearly increased the oocyte maturation rate compared to the control group (57 ± 2.42%), then it remained constant to the RJ40 (93 ± 3.10%) group. The higher RJ concentrations were associated with increased (p < 0.01) cleavage (53.3 ± 1.55% to 82.3 ± 2.82%) and blastocyst rate (15.5 ± 1.16% to 33.8 ± 3.09%) from the RJ0 to the RJ10 group. The relative mRNA expression of BCL2 and BAX in the oocyte was higher at RJ10. In cumulus cells, the expression of BCL2 was not affected, but that of BAX decreased, and expression of HAS2, PTGS2, and STAR were increased following the addition of RJ to the maturation media. In conclusion, the addition of 10 mg/mL of RJ to maturation medium improved blastocyst formation and decreased the apoptotic incidence in sheep cumulus cells and the oocyte during the in vitro development.

In this study, the effects of hydroalcoholic extract of red algae (Laurencia caspica) at different levels were evaluated on antioxidant defense, innate immune responses, and expression of immune genes in rainbow trout (Oncorhynchus mykiss). A total of 750 rainbow trout (39.43 ± 0.44 g) were randomly assigned to five treatments with three replicates (50 fish per replicate) in 15 ponds and reared for 8 weeks. Fish samples were fed a diet containing different levels of algal extract (0, 5, 10, 15, and 20 g extract per kg of basal diet). At the end of feeding period, the fish were challenged with Aeromonas hydrophila and fish mortality was recorded over a 10-day period. To evaluate the antioxidant and immune parameters, fish were sampled at day 0, end of weeks 4 and 8, and after bacterial challenge. Expression of immune genes (IL-1β, lysozyme II, Complement C3, and TNFα) was also examined after bacterial challenge. Results indicated that application of algal powder in fish diet improved lysozyme activity, complement system, and immunoglobulin levels of extract fed-fish compared to control group (p

Decline in semen quality is considered as a major contributing factor in age-related subfertility of broiler breeder flocks. This study was aimed to investigate the effect of dietary supplementation of Guanidinoacetic acid (GAA), as an alternative energy source along with antioxidant potential, on testicular histology and relative gene expression of some spermatogonial markers (c-Kit and STRA8) in aged roosters. Sixteen 24-week-old male broiler breeders were randomly allocated into four groups and fed a basal diet supplemented with increasing levels of GAA including 0 (GAA-0), 600 (GAA-600), 1200 (GAA-1200) or 1800 (GAA-1800) mg/kg diet/day for 26 successive weeks. At the end of the experiment, all the birds were killed and two ipsilateral testicle samples were taken to either quantify relative gene expression or do histology. Except for seminiferous tubules' diameter, testicular weight, and the number of blood vessels, dietary supplementation of GGA improved the epithelium thickness of seminiferous tubules, the number of spermatogonia and Leydig cells and the relative gene expression of c-Kit and STRA8 (P 0.01). Increasing levels of GAA cubically affected (P 0.01) the diameter of seminiferous tubules and their epithelium thickness as well as the number of spermatogonia. However, number of Leydig cells and relative expression of c-Kit were linearly, and relative expression of STRA8 was quadratically (P 0.01) enhanced in response to graded levels of GAA supplementation. Taking all parameters into account, daily supplementation of 1300-1450 mg of GAA/kg diet was estimated as an optimum dosage maximizing the evaluated traits.

The aim of this study was to investigate the effects of different dietary levels of L-arginine on the growth performance, blood parameters, and lipogenic gene expression of Arian broiler chickens. For this purpose, 168 Arian broiler chicks (40.33±1.7 g) were assigned to four treatments with three replicates of14 birds each, according to completely randomized design. The experimental treatments consisted of 100, 124, 139, and 154% dietary arginine levels relative to the published requirements of Arian broilers. On 42 d of the experiment, blood samples were collected from two birds (six birds per treatment) for blood metabolite measurements. These birds were then euthanized for carcass evaluation and collection of tissue samples. Increasing dietary arginine levels reduced (p

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.

Based on the findings of a recent study suggesting a decreased cold-induced ascites incidence in broiler progeny from hyperthyroid (HYPER) breeder hens, and a controversy on the effects of hyperthyroidism on immunocompetence, the present study was conducted to determine the probable adverse effect of induced maternal hyperthyroidism on immune function in progeny chicks. Breeder hens (n = 88) were randomly allotted to the control or HYPER groups and received common or thyroxine (T4)-added (1 mg/L) water, respectively. The hens were artificially inseminated, and hatching eggs (n = 924) were incubated. Thereafter, the male hatchlings (n = 288) were reared for 42 d, and several cellular and humoral immune responses were evaluated at standard or low ambient temperature. Prevaccination antibody titers to Newcastle disease, infectious bronchitis, and infectious bursal disease virus were higher in HYPER chicks during 1 wk of age, although not different in their dams. For primary response to SRBC administered at 7 d of age, HYPER chicks recorded higher total, IgM (d 14), and IgG (d 21) anti-SRBC antibody titers. Higher cutaneous basophilic hypersensitivity response in HYPER chicks (d 10) was not observed at 35 d of age. Carbon clearance assay showed no difference, but in vitro lymphoproliferative response to concanavalin A was higher in 19-d-old HYPER chicks, independent of temperature treatment. An increase in lymphocyte percentage coincided with a decreased heterophil percentage and heterophil to lymphocyte ratio (d 14) in the HYPER group. The weight of lymphoid organs in progeny was not influenced by the oral exposure of dams to extra T4. Independent of T4 treatment, cold exposure was generally associated with decreased immune functions at early stages. The data suggested that oral exposure of broiler breeder hens to 1 mg/L of T4 not only had no adverse effect on immune function, but also modulated early adaptive immune responses in progeny chicks for which the causal mechanisms remain to be unraveled.

Male broiler breeders (n = 32) of 55 weeks of age were administered four different doses of capsulated d-aspartate (DA; 0, 100, 200 or 300 mg kg−1 day−1, p.o. (DA0, DA100, DA200 and DA300 respectively)) for 12 successive weeks to assess reproductive performance, blood testosterone, testicular histology and transcript levels of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), androgen receptor (AR), LH receptor (LHR), 3β-hydroxysteroid dehydrogenase (3BHSD), proliferating cell nuclear antigen (PCNA), glutamate ionotropic receptor NMDA type subunit 1 (GRIN1) and glutamate ionotropic receptor NMDA type subunit 2B (GRIN2B). Blood samples and ejaculates were collected, and bodyweight was recorded weekly for 10 weeks. AI was performed weekly for the last 2 weeks to determine the number of sperm penetration holes in the perivitelline layer, fertility and hatchability. Testes histology and transcript levels were evaluated in the 12th week. Bodyweight, numbers of Leydig cells and blood vessels, testis index and levels of sperm abnormalities were not affected (P > 0.05) by the treatment. However, sperm total and forward motility, plasma membrane integrity and functionality of sperm, ejaculate volume, testosterone concentration and fertility were higher (P

The objectives of this study were to determine the effect of PGF2α treatment 14 d before the initiation of a G6G or G7G (PGF2α, 2 d, GnRH, 6 or 7 d, Ovsynch) protocol on ovarian response, synchronization protocol on ovarian response, progesterone (P4) concentration, pregnancy per AI (P/AI), and pregnancy loss in multiparous Holstein cows. Cows (3.6±1.1 lactations and yielding >30kg/d of milk) were randomly assigned to 1 of 4 timed artificial insemination (TAI) protocols: 1) G6G (n=240), one injection of PGF2α, GnRH 2 d later and a 7-d Ovsynch protocol (GnRH, 7 d, PGF2α, 56 h, GnRH 16 h TAI) was initiated 6 d later; 2) PG6G (n=250), PGF2α 14 d before the initiation of the G6G protocol; 3) G7G (n=200), one injection of PGF2α, GnRH 2 d later, and a 7-d Ovsynch protocol initiated 7 d later; and (4) PG7G (n=200), a PGF2α injection 14 d before the initiation of the G7G protocol. Blood samples from a subset of 269 cows were collected at the times of first and second GnRH, and PGF2α of the Ovsynch protocol to measure P4. Ultrasound examinations were performed to evaluate ovarian response to GnRH and PGF2α of Ovsynch, and to determine pregnancy status at 32 and 60 d after TAI. The proportion of cows with high (≥1ng/mL) P4 at first GnRH of Ovsynch was greater for PG6G and PG7G compared with G6G and G7G groups (combined 79.7 vs. 59.3%). In addition, mean (±SEM) plasma P4 concentration (ng/mL) at PGF2α of Ovsynch was also greater in PG6G (6.5±0.2) and PG7G (6.7±0.3) compared with G6G (5.1±0.2) and G7G (5.0±0.2). Cows given PGF2α 14 d before initiating a G6G or a G7G TAI (PG6G and PG7G) tended to have a greater P/AI at 32 d compared with those cows not receiving PGF2α (G6G and G7G). However, P/AI at 60 d was greater in cows subjected to PG6G and PG7G protocols (31.1 vs. 39.2%), with a lower pregnancy loss between 32 and 60 d (11.65 vs. 19.7%). In summary, administration of PGF2α 14 d before initiating a G6G or a G7G TAI protocol increased P4 concentrations before artificial insemination and late embryonic/early fetal survival in multiparous Holstein cows.

Probable involvement of avidin and avidin-related protein-2 (AVR2) in sperm viability in the sperm storage tubules of turkeys has been suggested. The high affinity of biotin to avidin and its analogs is also well documented. The present study aimed to determine the effect of oral biotin on reproductive performance and oviductal mRNA expression of avidin and AVR2 in 2 broiler hen lines with different fertility rates. Low-fertility (line B) and high-fertility (line D) hens (n=144) were randomly allotted to receive 0 (T0), 0.30 (T1), or 0.45 (T2) mg/L biotin in drinking water from 30 through 33 wk of age. The reproductive performance of the hens was evaluated using artificial insemination. At the end of the treatment period, 24 hens per line were killed to assay the expression of avidin and AVR2 in the uterovaginal junction. Supplementary biotin increased egg production from 73.5% for T0 to 87.8% for T2. Hens administered with biotin in line B, but not in line D, showed an increase (8.4%) in fertility rate. Hatchability, chick quality, and overall embryonic mortality were not different among the experimental groups. Real-time PCR data showed that both avidin (P=0.0013) and AVR2 (P

To study the effect of alpha-linolenic acid (ALA) on meiotic maturation, mRNA abundance of apoptosis-related (Bax and Bcl-2) molecules, and blastocyst formation in ovine oocytes., Purpose To study the effect of α-linolenic acid (ALA) on meiotic maturation, mRNAabundance of apoptosis-related (Bax and Bcl-2) molecules, and blastocyst formation inovine oocytes.Methods A preliminary experiment was conducted to analyze the concentration of ALAin "small" (≤2 mm) and "large" (≥6 mm) follicles using gas chromatography/massspectrometry analysis. The concentration of ALA in small and large follicles wasdetermined to be in a range of 75.4 to 125.7 μM, respectively. In vitro maturation (IVM)of oocyte was then performed in presence of 0 (control), 10 (ALA-10), 50 (ALA-50),100 (ALA-100), and 200 (ALA-200) μM of ALA. Meiotic maturation and mRNAabundance of Bax, and Bcl-2 genes was evaluated after 24 h of IVM. The embryoniccleavage and blastocyst formation following parthenogenetic activation were alsodetermined for each group.Results The highest concentration of ALA (ALA-200) decreased the oocyte maturationrate compared with the control group. Analysis of apoptosis-related genes in oocytesafter IVM revealed lesser transcript abundances for Bax gene, and higher transcriptabundances for Bcl-2 gene in ALA-treated oocytes as compared with the controloocytes. In term of cleavage rate (considered as 2-cell progression), we did notobserve any differences among the groups. However, ALA-100 group promoted moreblastocyst formation as compared with the control group.Conclusion Our results suggested that ALA treatment during IVM had a beneficial effect on developmental competence of ovine oocytes by increasing the blastocystformation and this might be due to the altered abundance of apoptosis-regulatorygenes.

Published data on the probable involvement of avidin and avidin-related protein-2 (AVR2) in sustaining sperm viability in sperm storage tubules in 38-wk-old turkeys, and the high affinity of avidin or its analogs to biotin suggest that supplementary biotin may increase oviductal avidin and AVR2 expression, thereby attenuating the adverse effect of aging on hen reproductive performance. Broiler breeder hens (n = 120) were randomly assigned to receive 0 (T-0), 0.30 (T-1), or 0.45 (T-2) mg of biotin/L of drinking water from 30 to 33 (young) and 53 to 56 (old) wk of age, and artificially inseminated to determine their reproductive performance. At the end of each period of biotin administration, 8 hens from each treatment group were killed for RNA extraction from the uterovaginal junction. Egg production was lower in the old hens (44%) compared with the young ones (82%), and biotin supplementation increased egg production only in the latter. Administering supplementary biotin to young hens increased their oviductal expression of AVR2, which was much higher in the old hens (1.0 and 4.6 for young and old groups, respectively). Fertility rate was not different between young and old hens, and was increased (4.4%) at the higher level of biotin supplementation. Hatchability and hatchling quality were not affected by biotin supplementation. Embryonic mortality between 17 to 21 d of incubation was higher in young (5.2%) compared with old (1.4%) birds. Egg fertility rate showed a moderate correlation (P < 0.05) with avidin (r = 0.59) and AVR2 (r = 0.55) expression in the young-age group, and very low correlations in old-age group (0.04 and 0.17). Regardless of the hen's age, the correlation coefficient of hatchability with avidin or AVR2 expression was very low (-0.16 and 0.18). Overall, the effect of biotin supplementation on AVR2 expression, and the relationship between biotin administration and oviductal expression of avidin and AVR2 was dependent on the hen's age, being higher in the young hens.

The aim of this study has been to determine the effects of in vivo post-ovulatory ageing (POA) on the distribution of spindle-associated proteins, histone H3/H4 post-translational modifications and on v-akt murine thymoma viral oncogene homolog 1 (Akt) expression levels. To this end, oocytes were retrieved 13, 29 and 33 h after human chorionic gonadotrophin (hCG) treatment. The presence and distribution at the meiotic spindle of acetylated tubulin, γ-tubulin, polo kinase-1 and Ser473/Thr308 phosphorylated Akt (pAkt) as well as histone H3 and H4 acetylation and phosphorylation levels were assayed via immunofluorescence. Akt expression levels were determined via reverse transcription–polymerase chain reaction and western blotting analyses. Spindles from oocytes recovered 13 h and 29 h after hCG treatment showed similar levels of acetylated tubulin but ageing induced: (1) translocation of γ-tubulin from spindle poles to microtubules, (2) absence of Thr308- and Ser473-pAkt in 76% and 30% of oocytes, respectively, and (3) a significant reduction in phosphorylation levels of serine 10 on histone 3. At 29 h, a significant decrease in Akt mRNA, but not in pAkt or Akt protein levels, was recorded. By contrast, protein content significantly decreased 33 h after hCG. We conclude that POA impairs oocyte viability and fertilisability by altering the expression levels and spindle distribution of proteins that are implicated in cell survival and chromosome segregation. Together, these events could play a role in oocyte apoptosis.

A hypothesis was tested that the in ovo injection of biological buffers may reinforce the buffering capacity of albumen, thereby withstanding the increase in albumen pH during storage and improving hatchability and chick quality in long-term stored eggs. Hatching eggs (n = 2,420) were randomly assigned to 11 treatment groups (4 replicates of 55 eggs each) and injected (d 1) with distilled water, 25 or 50 mM HEPES (H25 and H50), Bicine (B25 and B50), Tris (T25 and T50), and Bis-Tris-propane (BTP25 and BTP50) solutions or were not injected (intact: control; or pricked with a needle: N). The eggs were then stored for 14 d during which the egg internal characteristics were evaluated at 1, 2, 3, 4, 5, 8, and 13 d of storage (n = 924 in total) and the remaining eggs (n = 1,496) were incubated. A decrease in albumen pH was found for H25, H50, B50, and BTP25 groups from 2 through 5 d postinjection. Eggs receiving H25, H50, and B50 recorded a higher albumen index (at 13 d of storage) and Haugh unit (between 8 and 13 d of storage) compared with the control. Interestingly, the hatchability of fertile eggs was influenced by the treatment effect (P = 0.0001) where the eggs receiving H25 (88.3%), H50 (88.9%), B50 (88.4%), and BTP25 (87.6%) recorded higher values than that of control (82.1%), associated with a decreased early embryonic mortality rate (P0.0001). In ovo injection of Tris buffer, however, profoundly decreased the hatchability (47.2 and 29.0% for T25 and T50, respectively) and percentage of first-grade chicks (67.5 and 63.6% for T25 and T50, respectively) compared with the control (90.1%). In conclusion, prestorage in ovo injection of H25, H50, B50, and BTP25 improved hatchability in long-term stored eggs in which a decreased albumen pH during the d 2 through 5 of storage period might be involved.

A hypothesis was tested that providing the breeder hens with exogenous thyroxine (T(4)) would help their offspring to better survive the ascites-inducing condition during the growing period. In total, 132 broiler breeder hens were randomly assigned to one of 3 treatments: control (CON), hypothyroid [HYPO; 6-N-propyl-2-thiouracil (PTU)-treated], and hyperthyroid (HYPER; T(4)-treated). The hens were artificially inseminated, and the hatching eggs (n = 1,320) were incubated. No eggs in the HYPO group hatched. The 1-d-old male chicks (n = 288) from other groups were reared for 42 d under standard or low ambient temperature to induce ascites. Blood samples were drawn from the hens, embryos, and broilers for determination of T(4) and triiodothyronine (T(3)). The hematocrit was also determined in broilers. The PTU-treated hens had an increased BW along with lower plasma T(3) and T(4) concentrations. Plasma T(4) was higher in the HYPER hens compared with CON hens, but T(3) concentration was not different between these groups. The fertility rate was not affected by either hypo- or hyperthyroidism. The embryos in the HYPO group had lower plasma T(3) and T(4) concentrations at d 18 of embryonic development and internal pipping. Higher plasma T(4) was recorded in the HYPER birds at internal pipping, although plasma T(3) concentration was not affected at this stage. Maternal hyperthyroidism decreased the overall incidence of ascites in the cold-exposed chickens (10.0 vs. 33.4% for HYPER and CON groups, respectively). Although the effect of maternal PTU or T(4) treatment on plasma thyroid hormones and on the right ventricle-to-total ventricular weight ratio in the broilers was not significant, the cold-exposed healthy CON chicks showed higher hematocrit values, compared with the HYPER birds. It was concluded that maternal hyperthyroidism could decrease the incidence of cold-induced ascites in broiler chickens; however, probable causal mechanisms remain to be elucidated.

To determine whether G6PDH-activity measured by Brilliant Cresyl Blue known as BCB dye, predicts developmental competence within cohorts of ovine oocytes.Ovine oocytes were exposed to BCB staining and categorized into two groups: BCB+ (blue cytoplasm, low G6PDH-activity) and BCB- (colorless cytoplasm, high G6PDH-activity). After maturation in vitro, oocytes were subjected to fertilization followed by in vitro embryo culture.We observed a significant difference in oocyte diameter considering BCB+ and BCB- oocytes. BCB+ and Control groups showed significantly higher maturation rates compared to BCB- group. There were significantly more cleaved embryos in BCB+ and control groups than in BCB- group. Blastocyst rate was significantly higher for BCB+ group compared to control and BCB- groups with control group being significantly higher than BCB- group.G6PDH-activity is a strong predictive marker of oocyte competence and may be useful in identifying oocytes with a good prognosis for further develop.

Nine sexually mature rams, three Shall, three Afshari and three Zandi were used in this study. Semen of rams was collected twice per month during the four seasons of the year and blood samples were obtained via jugular vein. Seminal characteristics, scrotal dimension and blood parameters were measured at 2 week intervals. Time of the year significantly (p < 0.05) affected the volume of semen, sperm concentration, percentage live sperm, percentage abnormal sperm, sperm mass motility, total sperm number, scrotal circumference, relative testis volume and serum levels of testosterone, triiodothyronine and thyroxine. All of semen characteristics, except sperm abnormality, were high in summer months than at other months of the year (p < 0.05). Sperm abnormality was higher in autumn months (p < 0.05). Relative testis volume and testicular circumference were higher in autumn and summer months than at other mouth of the year, respectively (p < 0.05). Scrotal circumference were not significant between Shall and Afshari breeds but comparison with Zandi breed, Scrotal circumference was bigger (p < 0.05). The largest values for thyroid hormones synchronized with low reproductive performance of rams, but testosterone was highest levels at this time (p < 0.05). TimexBreed interaction effect was significant for sperm concentration, total sperm number, semen volume, testis circumference, relative testis volume, thyroid hormones and testosterone (p < 0.05).

Royal jelly (RJ) is a secretion product from the cephalic glands of nurse bees that has extraordinary properties and remarkable health effects. Over the years, antioxidative and antiapoptotic properties of RJ have been experimentally investigated. Here we hypothesised that supplementary RJ in in vitro maturation (IVM) medium would (i) improve cumulus expansion (ii) oocyte nuclear maturation, (iii) glutathione (GSH) content, and (iv) mitochondrial activity, and (v) also affect the mRNA abundance of the (Bax, Bcl-2, and p53) transcripts involved in oocyte apoptosis. To test these hypotheses, goat ovaries were collected from a local abattoir and transported to the laboratory. Cumulus-oocyte complexes (COC) with multilayered compact cumulus investment and evenly granulated cytoplasm were selected and randomly allocated to the experiments. To evaluate the effects of RJ on meiotic competence after maturation in vitro, IVM medium was supplemented with concentration of 0.0 (RJ-0), 2.5 (RJ-2.5), 5.0 (RJ-5), and 10.0 (RJ-10) mg mL–1 of RJ. After IVM, oocytes of each group were evaluated for cumulus expansion (visual assessment), stage of nuclear maturation (Hoechst staining), intracellular level of GSH (Cell Tracker blue staining), mitochondrial activity (MitoTracker Deep Red staining), and relative expression of Bax, Bcl-2, and p53 genes (qRT-PCR assay). Differences were analysed for significance by one-way ANOVA using SAS version 8.0 (SAS Institute Inc., Cary, NC, USA), considering P 0.05). Our results revealed that maturation rate was higher (88.0%) in the RJ-10 group when compared with the RJ-2.5 (71.5%) and control (RJ-0) groups (60.0%; P 0.05). A higher (P 0.05) was observed among RJ-treated and untreated oocytes regarding their mitochondrial activity after IVM. Based on these results, the concentration of 10 mg mL–1 (RJ-10) was selected for evaluation of Bax, Bcl-2, and p53 transcripts abundance. Our results revealed that the expression of Bax mRNA was decreased (P 0.05). In conclusion, results of this study showed that adding RJ to maturation medium at optimum concentration increased the nuclear maturation and GSH synthesis, but not activity of the mitochondria; this improvement was associated with expression of apoptosis-related genes in goat oocytes.

In this experiment, four hundred day-old broiler chicks were assigned randomly to 16 floor pens with 25 chicks each. Three diets containing 0% fish oil + 7% soybean oil, 3.5% fish oil + 3.5% soybean oil and 7% fish oil + 0% soybean oil and a free oil control diet were formulated. The birds fed fish oil diet showed a lower body weight than control group (P < 0.05). Abdominal fat percent in birds fed soybean oil and fish oil diets, were lower than control group (P < 0.05). The high density lipoprotein (HDL) concentration in birds fed fish oil diet was higher than other treatments, but the serum low density lipoprotein (LDL) decreased in birds fed fish oil diet (P < 0.05). The live weight of birds was positively correlated with glucose and LDL and negatively correlated with HDL concentrations (P < 0.01). The abdominal fat pad percent was positively correlated with triglyceride, glucose, LDL and very low density lipoprotein (VLDL) and a negative correlation was observed with HDL concentrations (P < 0.01). Triglyceride, cholesterol and VLDL concentrations were positively correlated with each other and a negative correlation was observed between high-density lipoprotein and low-density lipoprotein (P < 0.01). The results of the present study showed that higher fish oil inclusion in broiler diets can result in some unfavorable reduction in live weight and a beneficial influence on abdominal fat reduction.Keywords: Broiler chickens, fish oil, high-density lipoprotein, low-density lipoproteinbr>African Journal of Biotechnology Vol. 9(35), pp. 5779-5783, 30 August, 2010

Different mutations in the bone morphogenetic protein-15 (BMP-15) and the Growth Differentiation Factor-9 (GDF-9) genes have increased ovulation rate and infertility in a dosage-sensitive manner insheep. To test the polymorphisms of genes in goat, which have been demonstrated as major genes of fecundity in sheep, the genetic polymorphism of FecXB and FecXG loci in BMP-15 gene were studied in109 Iranian native goats. Blood samples were collected in EDTA coated tubes from Jugular vein and genomic DNA was extracted from whole blood samples. Single nucleotide polymorphism of FecXB and FecXG loci in BMP-15 gene were determined using PCR-RFLP technique. There was no evidence of mutation in FecXB and FecXG in these goats, all of which were monomorph for exon 2 BMP-15 gene.