Calcitriol combined with fish oil in the diet of Chukar partridge can ameliorate laying performance by improving oviduct histomorphometry and functions.

The effects of calcitriol (CT) and/or fish oil (FO) on performance, oviposition time, sex ratio and morphology of the reproductive system of laying Chukar partridges were studied. Female (n = 48) and male (n = 16) partridges were used in a completely randomised design using a 2 × 2 factorial arrangement and were randomly allocated to either of four experimental treatments with four cage replicates of three females and one male each. Female birds received no FO (CON - FO) or were orally administered with 0.2 mL (0.24 g)/500 g body weight FO (CON + FO) or 0.2 mL solution containing 10 µg CT (CT - FO), or their combination (CT + FO) for 42 successive days. The eggs were collected every two hours between 07:00 and 23:00 h. Administering FO along with CT had considerably increasing effect on the male-biased sex ratio. FO and CT administration interacted to increase serum calcium concentration. Experimental treatments increased the number of leucocytes and erythrocytes. Serum cholesterol was decreased in CON + FO partridges compared with those of the CT - FO and CON - FO birds. There was an interaction between FO and CT on the weight of eggs and hatchlings, number of medium white follicles, diameter and the number of small yellow follicles, the weight and diameter of the fifth follicles (F5), and thickness of secondary mucosal folds in both uterus and vagina. Administering CT alone or with FO increased the feed intake, egg production, oviductal weight, diameter and number of large yellow follicles, the weight and diameter of the second (F2) and first (F1) follicles compared with those of the CON - FO females. Further studies are required to elucidate the mechanisms by which such changes in the sex ratio skew, ovary and oviduct are mediated.

Melatonin as a Smart Protector of Pregnancy in Dairy Cows.

The experimental objective was to examine the role of melatonin and its pathways in the maintenance of pregnancy in lactating dairy cows. Blood samples were collected at days 0, 16 and 32 after timed AI from cows (n = 200) in order to consider plasma melatonin concentrations and to conduct AOPP (advanced oxidation products of proteins) and TBARS (thiobarbituric acid reactive substances) tests. Luminal endometrial cells were collected at day 16 using a Cytobrush in all cows to determine mRNA expressions of melatonin receptor 1 (MT1), mouse double minute 2 (MDM2), MDM2 binding protein (MTBP), BCL2-associated X, apoptosis Regulator (BAX), p53 upregulated modulator of apoptosis (PUMA, gene symbol BBC3), mucin 1 (MUC1) and leukemia inhibitory factor (LIF). Plasma concentrations of melatonin were significantly greater in pregnant cows diagnosed pregnant at day 16 who sustained pregnancy to day 32 compared to nonpregnant cows at day 16, or pregnant at day 16 and who lost embryos by days 32. Concentrations of AOPP and TBARS were greater in nonpregnant cows at day 16 or pregnant at day 16 and who lost embryos by days 32 compared to those diagnosed pregnant at day 16 and who sustained pregnancy to day 32. In pregnant cows, endometrial mRNA expressions of MDM2, MTBP, MTR1 and LIF were higher compared to pregnant-embryo-loss cows (p < 0.05). In contrast, mRNA expressions of BBC3 and MUC1 were greater at day 16 in pregnant-embryo-loss cows compared to pregnant cows (p < 0.05). In conclusion, melatonin status is a modulator of embryo well-being and maintenance of pregnancy in lactating dairy cows.

Royal jelly may improve sperm characteristics during preservation of rooster semen: Gene expression of antioxidant enzymes.

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 < .05) the percentages of forward progressive motility, viability and HOST during liquid and frozen storage. The abnormality of spermatozoa in the RJ5 group was significantly lower compared to the other groups. During liquid storage, a significant decrease in forward progressive motility was found after 48 hr in comparison with 24 hr at 4°C. High levels of RJ (from 10 to 40 mg/ml) were severely decreased the characteristics of rooster spermatozoa in comparison with RJ5 and the control group. The inclusion of RJ at 5 mg/ml to the semen extender enhanced the mRNA transcript of antioxidant enzymes of spermatozoa during liquid preservation. The mRNA abundance of antioxidant enzymes did not influence by cryostorage. Overall, these data suggest that supplementation of RJ at 5 mg/ml to the extender improved semen characteristics and redox status of rooster spermatozoa.

Detrimental effects of long-term exposure to heavy metals on histology, size and trace elements of testes and sperm parameters in Kermani Sheep.

Exposure to endocrine-disrupting chemicals (EDCs) has been hypothesized as a cause of declining sheep reproductive efficiency. Understanding the long-term effects of EDCs such as heavy metals on reproductive health requires investigation in 'real life' of sheep that are reared in industrial areas. The aim of this study was to evaluate the effect of long-term exposure of Kermani rams to high levels of environmental heavy metals probably emitted from a copper smelter at KhatoonAbad in ShahreBabak, Kerman province. Testicular characteristics were determined in randomly-selected rams (3-4 years old) at 4 directions (south, north, east, and west) and 4 distances (10, 20, 30, and 40 km) from the smelter. Testicular trace element contents, size, serum testosterone, histological attributes and seminal characteristics, except semen volume, were affected by both the direction and the distance from the smelter (P < 0.05). Testicular contents of Pb, Cd, Cr, and Ni, and sperm abnormalities were higher at 10 km south from the smelter and lower at 40 km west. Other parameters were higher at 40 km west and lower at 10 km south. Interestingly, the testicular contents of Cu at 10 km south were lower and associated with higher sperm abnormalities in the rams reared closer to the smelter. The highest weight, length and circumference of the testis were found at 40 km west. The lowest concentration of testosterone was observed at 10 km south, being 92.6% lower than the highest values obtained at 40 km west. The diameter of seminiferous tubules and epithelial height at 10 km south were 8.9% and 27.5% lower than the highest values obtained at 40 km west. A positive correlation between Pb, Cd, Cr and Ni contents in the testis with sperm abnormalities, and a negative correlation between these elements with the other parameters were found. It was concluded that long-term exposure to heavy metals might have been a cause of decreased fertility in rams and probably other living species in this region.

Oral administration of royal jelly may improve the preservation of rooster spermatozoa.

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.

Association of CYP1A1 M2 (A2455G) Polymorphism with Susceptibility to Breast Cancer in Mazandaran Province, Northern Iran: A Case-control Study.

BACKGROUND: Breast cancer is one of the most frequent women malignancies in the world. The cytochrome P450 1A1 (CYP1A1) is a key enzyme in xenobiotics metabolism. Moreover, CYP1A1 plays a critical role in the etiology of breast cancer by involving in 2-hydroxylation of estrogen. Therefore, single-nucleotide polymorphisms (SNPs) of its coding gene have been verified to be important in cancer susceptibility. The aim of the study was to evaluate the association of CYP1A1 M2 (A2455G) includes rs1048943 of this SNP polymorphism with the risk of breast cancer in Mazandaran province. METHODS: Ninety-six breast cancer patients with known clinicopathological characters and 110 healthy women as control were genotyped for CYP1A1 M2 polymorphisms by the restriction fragment length polymorphism technique. RESULTS: The analysis of CYP1A1 gene (polymorphism M2) showed that the frequency of homozygous wild genotypes (AA), heterozygous (AG), and mutant genotype (GG) in the patient group, respectively, 78%, 22%, and 0%, and also the frequency of genotypes AA, AG, and GG in healthy included 82%, 16%, and 2%, respectively. Statistical analysis by Logistic regression model at P < 0.05 showed no significant correlation between polymorphisms in CYP1A1M2 and breast cancer risk (odds ratio = 0.84, confidence interval = 0.33-2.17). CONCLUSIONS: The results indicated that the M2 allelic genotypes were significantly associated neither with breast cancer risk nor with clinicopathological characteristics in Mazandaran province.

Antioxidant levels, copper and zinc concentrations were associated with postpartum luteal activity, pregnancy loss and pregnancy status in Holstein dairy cows.

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 ±â€¯0.25, mean ±â€¯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.

Polymorphism detection of promoter region of IFN-γ and IL-2 genes and their association with productive traits in Mazandaran native breeder fowls.

To identify polymorphism in interferon gamma (IFN-γ) and interleukin-2 (IL-2) genes, blood samples were collected from 380 breeder hens of the Mazandaran native fowls breeding station. DNA extraction was performed through a modified saltingout method and fragments of 670 and 659 bp from the promoter regions of IFN-γ and IL-2 genes were amplified by using specific primers, respectively. Following genotyping in the IFN-γ gene using the Tsp509I restriction enzyme, two alleles of A and G with the frequencies of 0.55 and 0.45 and three genotypes of AA, AG and GG were observed with the frequencies of 0.32, 0.46 and 0.22, respectively. For the IL-2 gene, two alleles of A and G were also detected using the MnlI restriction enzyme with the frequencies of 0.58 and 0.42 and three genotypes of AA, AG and GG with the frequencies of 0.33, 0.50 and 0.17, respectively. Statistical analysis revealed significant associations between IL-2 gene single-nucleotide polymorphism and productive traits including the average egg weight (EW) at 345-375 days of age, egg number (EN) at 345-375 days of age and body weight (BW) at 8weeks of age traits (P<0.05). Further, in a mean comparison analysis, there were also significant differences between different genotypes of the IL-2 gene in average EWat 28 and 30weeks of age, in which AG genotypes showed higher performance. Additionally, for the IFN-γ gene, a significant difference was found between the genotypes in average EW at 28 weeks of age trait. Therefore, it can be concluded that the above-mentioned polymorphisms could be considered as the pivotal geneticmakers to improveMazandaran native fowl breeding programmes to achieve the optimum performance in productive traits more efficiently.

Effect of cyclical cold stress during embryonic development on aspects of physiological responses and HSP70 gene expression of chicks.

The objective of the present study was to evaluate the effects of cyclical lower incubation temperature at different embryonic ages on the hatchability, body and organs weights, thyroid hormones, and liver HSP70 gene expression of newly hatched chicks. In a completely randomized design, fertile eggs of a broiler breeder (34 weeks of age) were assigned to three treatment groups with six replicates and 145 eggs per each. The treatment groups were as: control group (C) that eggs were incubated at 37.6°C during the whole incubation period; incubation temperature was decreased to 36°C for 3h per day at embryonic age from 12 to 14 (T1); and incubation temperature was decreased to 36°C for 3h per day at embryonic age from 15 to 17 (T2). No significant difference was found among treatments for hatchability (P>0.05). There were no differences (P>0.05) among treatments for body weight and liver weight, while heart weight of chicks in T1 and T2 groups were significantly higher than the control group (P<0.05). There were no differences (P>0.05) among treatments for the levels of thyroid hormones, however, the levels of both hormones tended to increase in chicks exposed to cold stress (T1 and T2). Chicks in T2 group had higher liver HSP70 gene expression compared with those in T1 and the control group (P<0.05). Cold stress in both incubation periods had no significant effect on the plasma levels of aspartate aminotransferase and alanine aminotransferase. Treatments had no effect on the plasma levels of glucose, cholesterol and triglyceride. The results of this study suggest that cyclical lower incubation temperatures (36°C) at the embryonic age from day 15-17 could induce the liver HSP70 gene expression, without negative effects on the hatchability and body weight of hatched chicks.

Impact of supplementary royal jelly on in vitro maturation of sheep oocytes: genes involved in apoptosis and embryonic development.

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.