Investigation of the 5' flanking region and exon 3 polymorphisms of IGF-1 gene showed moderate association with semen quality in Sanjabi breed rams.

In this study, semen samples were collected from 96 Sanjabi rams in order to investigate the IGF-1 gene polymorphisms and their relationship with the characteristics of semen quality and testicular size. The dimensions of scrotal length, width and circumference were measured during autumn and spring over two years. Blood samples were simultaneously collected from jugular vein to extract DNA. PCR was performed using specific primers to amplify 294 and 272bp fragments including 5' regulatory region and exon 3 of IGF-1 gene, respectively. PCR products were digested by BFOI and Eco88l restriction enzymes, respectively. Two genotypes including AA (194 and 100bp), AB (294, 194 and 100bp) and all possible genotypes including CC (182 and 90bp), CT (272, 182, and 90bp) and TT (272bp) were observed for 5' flanking region and exon 3 of IGF-1 gene, respectively. The significant differences among IGF-1 genotypes for testicular dimensions were not observed. However, the polymorphism of 5' flanking region in the studied population had significant effect on individual motility and percent morphology traits. Animals with AB genotype had significantly higher individual motility compared with AA genotype (P < 0.05). Also, animals with AA genotype had significantly the highest percent morphology compared with AB genotype (P < 0.1). The exon 3 of IGF-1 gene had significant effect on individual motility, concentration, morphology and water test traits. Animals with CT genotype had the highest sperm concentration (P < 0.1) and water test (P < 0.05) compared to CC and TT genotypes. Moreover, animals with TT genotype had significantly the highest percent morphology compared with other genotypes (P < 0.05). Briefly, the results indicated that individual motility, concentration, percent morphology and water test traits could be in association with IGF-1 genotypes. It might be concluded that polymorphisms in IGF-1gene can be considered to develop male fertility in future and for using in selection process of better animals under masker assisted selection programs.

Effect of sericin supplementation in maturation medium on cumulus cell expansion, oocyte nuclear maturation, and subsequent embryo development in Sanjabi ewes during the breeding season.

The purpose of this study was to evaluate the effect of sericin with different concentrations (0% [control], 0.1%, 0.5%, 1.0%, and 2.5%) added to the IVM medium on cumulus cell expansion, oocyte nuclear maturation, and subsequent embryo development in Sanjabi ewes during the breeding season. The resumption of meiosis was assessed by the frequency of germinal vesicle breakdown and the first polar body extrusion. After IVF with fresh ram semen, presumptive zygotes were cultured 8 days in potassium simplex optimization medium supplemented by amino acids, and the percentages developing to the two-cell and blastocyst stages were measured as the indicators of early embryonic developmental competence. More cumulus-oocyte complexes matured with 0.5% sericin underwent germinal vesicle breakdown and reached metaphase II stage compared with the control cumulus-oocyte complexes matured without sericin (P ≤ 0.05). The present findings indicated that supplementation with 0.5% sericin during the maturation culture may improve the nuclear maturation and the cumulus cell expansion. Furthermore, the percentage of blastocysts obtained from 0.5% and 0.1% sericin (37.8 ± 1.76% and 34.8 ± 1.09%, respectively) was higher (P ≤ 0.05) than that of the control medium (29.60 ± 1.67%). However, addition of 1% and 2.5% of sericin to the IVM medium oocytes had a negative effect on nuclear maturation and cumulus cell expansion. Furthermore, the percentage of cleavage and blastocyst rate was significantly lower in the 1% and 2.5% sericin groups than in the control group. These findings showed that supplementation of IVM medium with 0.5% sericin may improve the meiotic competence of oocytes and early embryonic development in Sanjabi ewes during the breeding season.

Effect of PGF2α and GnRH on the reproductive performance of postpartum dairy cows subjected to synchronization of ovulation and timed artificial insemination during the warm or cold periods of the year.

This study was designed to evaluate the reproductive performance of lactating dairy cows (Holstein Friesian) after the injection of PGF2α analogue on Day 15 postpartum, and GnRH analogue on Day 23 after artificial insemination (AI) with Presynch (two injections of PGF2α, administered 14 days apart starting at 30-35 days postpartum) + Ovsynch-based (GnRH-7 days-PGF2α-2 days-GnRH-16-20 hours-timed artificial insemination) treatments, during the warm and cold periods of the year. All the cows (n = 313) were assigned to one of the four groups including: M1 (n = 72) in which the cows were treated with PGF2α on Day 15 postpartum + Presynch-Ovsynch + GnRH on Day 23 post-AI; M2 (n = 41) in which the cows received PGF2α on Day 15 postpartum + Presynch-Ovsynch; M3 (n = 100) including the cows that got Presynch-Ovsynch; and control group (n = 100) including the cows that were not treated and were inseminated at natural estrus. Pregnancy diagnosis was performed 28 to 35 days post-insemination by means of ultrasound. The results showed that treatment with PGF2α on Day 15 postpartum significantly decreased the days to conception and the number of services per conception (P < 0.01) and it also improved the first service conception rate (P < 0.1) only in cows that were treated with M2 protocol. Whereas, the days to first service was not influenced by the treatment of PGF2α on Day 15 postpartum (P > 0.05). In contrast, administration of GnRH on Day 23 post-AI increased the days to conception and the number of service per conception (P < 0.01) and tended to decrease the first service conception rate (P < 0.1) in cows that were treated with M1 compared with M2 protocol. Therefore, it was concluded that Presynch-Ovsynch protocol could be more reproductive and beneficial when a single treatment with PGF2α was administered at 15 days postpartum (15 days after the PGF2α, Presynch-Ovsynch protocol was initiated). Interestingly, the administration of a GnRH agonist on Day 23 post-AI not only did not improve the reproductive performance of the cows receiving first postpartum timed artificial insemination after Presynch-Ovsynch protocol but also reduced that.

SNP exploring in the middle and terminal regions of the IGF-1 gene and association with production and reproduction traits in Holstein cattle.

Five primer sets were designed in order to identify single nucleotide polymorphisms (SNPs) in middle and terminal exons (2 to 6) and in some flanking intronic regions of the bovine insulin-like growth factor 1 (IGF-1) gene. Sequencing results of PCR products for 10% of animals showed no variant in exons but a SNP at intron 4 was occurred. Both polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and high resolution melting (HRM) methods were developed to genotype samples. The PCR-RFLP results showed the presence of three fragments on agarose gel for the C allele due to two cleavage sites while two fragments for the T allele were observed. Melting curves of 123bp fragments in HRM analysis showed a difference between temperature melting (Tm) of two homozygous genotypes as the CC genotypes had higher Tm than the TT genotypes. Melting curve of the CT genotype was different and crossed two parallel patterns of homozygous genotypes. The frequencies of the CC, CT and TT genotypes were 0.6, 0.37 and 0.03, respectively. Also, the estimated allele frequencies were 0.785 and 0.215 for the C and T alleles, respectively. Results showed higher accuracy of the HRM analysis compared to the PCR-RFLP method. Least square means (LSMs) comparison of the different genotypes in the SNP showed significant association with milk fat yield trait in the first lactation and open days after the second calving. The polymorphism did not have a significant effect on other milk production or reproduction traits. It seems that other variants or QTLs known in this region underlie genetic variation in the production and reproduction of dairy cattle.

Combination of oocyte and zygote selection by brilliant cresyl blue (BCB) test enhanced prediction of developmental potential to the blastocyst in cattle.

The cumulus oocyte complexes (COCs) were obtained from local abattoir. After aspiration, the COCs were allotted into four treatments to evaluation of brilliant cresyl blue (BCB) test. Control treatment (C): oocytes were cultured directly (without exposure to BCB) after recovery in in vitro production (IVP) process. Oocyte treatment (OBCB): immediately after aspiration, COCs were incubated in modified Dulbecco's phosphate-buffered saline (mDPBS) supplemented with 26µM of BCB for 90min and classified into two classes: oocytes with blue cytoplasm coloration (OBCB+: more competent oocytes) and oocytes without blue cytoplasm coloration (OBCB-: low competent oocytes). Directly after classification, the oocytes were maintained undisrupted in the IVP process. Zygote treatment (ZBCB): After oocyte collection, maturation and fertilization, zygotes were stained with BCB for 10min and categorized into three ways, according to whether they were highly stained (ZBCB++: low competent zygotes), moderately stained (ZBCB+: moderate competent zygotes) and unstained (ZBCB-: more competent zygotes). Directly after classification, the zygotes were maintained undisrupted in the culture process. Oocyte and zygote treatments (OBCB/ZBCB): COCs were stained with BCB after recovery and classified into two classes (OBCB+ and OBCB-). After fertilization, the zygotes produced from OBCB+ and OBCB- oocytes were further stained with BCB for 10min and categorized six ways (OBCB+/ZBCB++, OBCB+/ZBCB+, OBCB+/ZBCB-, OBCB-/ZBCB++, OBCB-/ZBCB+ and OBCB-/ZBCB-). Directly after classification, the zygotes were maintained undisrupted in the culture process. The selection rate produced from OBCB treatment (OBCB+; 54.3%) was greater (P<0.05) than ZBCB treatment (ZBCB-; 44.3%). In addition, the selection rate produced from double application (combination of oocyte and zygote selection) of BCB test (OBCB+/ZBCB-: 28.8%) was less (P<0.01) than single application of BCB test (ZBCB-: 44.3%or OBCB+: 54.3%). The percentage of blastocyst production from OBCB+ oocytes (35.7%) and ZBCB- zygotes (36.6%) were greater (P<0.05) than that from C oocytes (25.7%), OBCB- oocytes (16.5%), ZBCB++ (13.5%) and ZBCB+ zygotes (21.3%). However, there were no significant differences (P>0.05) in the percentages of blastocyst production between OBCB+ oocytes (35.7%) and ZBCB- zygotes (36.6%). The proportion of blastocyst production from double application of BCB test (OBCB+/ZBCB-: 48.0%) was greater (P<0.05) than that from single application of BCB test (OBCB+: 35.7% or ZBCB-: 36.6%). In conclusion, current results confirmed that combination of oocyte and zygote selection by BCB test enhanced the efficiency of selecting for high quality embryos, compared to the single BCB test.

Using an incomplete gamma function to quantify the effect of dystocia on the lactation performance of Holstein dairy cows in Iran.

The aim of the present study was to estimate the effect of dystocia on lactation performance, using an incomplete gamma function. Data from March 2000 to April 2009 comprising 100,628 lactations of 65,421 cows in 204 dairy herds collected by the Animal Breeding Center of Iran were used. Of 100,628 births, 91.8% required no assistance, whereas 8.2% required assistance of some sort. Factors associated with the presence of dystocia were calving season, calving year, herd, calf sex, parity, and age of dam. Peak yield for primiparous cows with dystocia at calving occurred on d 87.2 [standard error (SE) 0.47], and for primiparous cows with easy calving, the peak of lactation was on d 83.3 (0.25). Peak yield was lowered by 0.39 (SE 0.07), 2.20 (SE 0.15), 2.22 (SE 0.21), and 2.54 (SE 0.32) kg for cows with incidence of dystocia compared with normal cows in parity 1 to 4, respectively. Dystocia was associated with decreased 305-d lactation performance in all parities, mostly in early lactation. Although more difficult births occurred in heifer calvings, loss in lactation performance was greater in second or later lactations following a difficult birth.

Effects of different levels of organic and inorganic chromium on growth performance and immunocompetence of broilers under heat stress.

This experiment was carried out to investigate the effects of different levels of organic and inorganic chromium on the performance, immune function and some serum mineral concentrations of broilers under heat stress condition (23.9-37°C cycling). A total of 150 one-day-old broiler chicks according to a completely randomized design were assigned into five treatment groups. Each treatment consisted of three replicates and each replicate contained ten chicks. Chicks were fed on corn-soybean meal basal diets with added different concentrations of chromium (0, 600 and 1,200 µg kg(-1) chromium chloride or 600 and 1,200 µg/kg chromium L-methionine) from 1 to 49 days of age. Humoral immunity was assessed by intravenous injection of 7% sheep red blood cell (SRBC) followed by evaluation of serum for antibody titers in primary and secondary responses. Cell-mediated immunity was assessed by the cutaneous basophil hypersensitivity (CBH) test to phytohemagglutinin (PHA)-P at day 32 and PHA-M at day 48. Heterophil/lymphocyte (H/L) ratio was also measured as a reliable indicator of stress. The body mass, feed intake and conversion ratio were not influenced by dietary chromium (P > 0.05). Dietary supplementation of both organic and inorganic chromium significantly increased primary and secondary antibody responses (P < 0.01), and also improved H/L ratio (P < 0.05), CBH response (P < 0.01) as well as relative weights of thymus (P < 0.05) and spleen (P < 0.01). Both dietary organic and inorganic chromium caused an increase in serum concentrations of Cr and Zn (P < 0.01), but decreased the serum concentration of Cu (P < 0.01). These results suggest that supplemental chromium especially in organic form offers a good management practice to reduce heat stress-related depression in immunocompetence of broiler chicks.