Addition of Recombinant Human Growth Hormone to In Vitro Maturation Medium of Bovine Oocytes

The aim of this study was to increase the bovine embryonic development rate, adding recombinant human growth hormone (rhGH) to maturation medium of bovine oocytes. Oocytes were matured for 24 h in TCM 199 Earle's salts and five treatments were developed: T1, 0.01 IU/ml of recombinant human follicle stimulating hormone (rhFSH); T2, 0.01 IU/ml of rhFSH + 100 ng/ml of rhGH; T3, 0.01 IU/ml of rhFSH + 1000 ng/ml of rhGH; T4, 100 ng/ml of rhGH; and T5, 1000 ng/ml of rhGH at 39 degrees C and 5% of CO(2) in air and saturated humidity. In vitro fertilization from cumulus-oocyte complexes was conducted in TALP-Fert medium (18-22 h) and spermatozoa were selected by Percoll gradient. Zygotes were incubated in SOFaaci medium in 5% of CO(2) in air, 5% of O(2) at 39 degrees C and saturated humidity for 11 days. There was no statistical difference in cleavage rate and embryo production on day 7 and day 9 among treatments. However, the hatching rate increased significantly in the T4 and T5 treatments (11.0 and 12.8%, respectively), compared with the T1 treatment (4.6%) (p < 0.05). Therefore, the rhGH addition to the oocyte maturation medium showed beneficial effects on the hatching rate of in vitro-produced bovine embryos.     

Influence of nutrient intake and body fat on concentrations of insulin-like growth factor-I, insulin, thyroxine, and leptin in plasma of gestating beef cows

Pregnant Angus x Hereford cows (n = 73) were used to determine the effects of amount of nutrient intake and BCS on concentrations of IGF-I, insulin, leptin, and thyroxine in plasma. At 2 to 4 mo of gestation, cows were blocked by BCS and assigned to one of four nutritional treatments: high (H = a 50% concentrate diet fed ad libitum in a drylot) or adequate native grass pastures and one of three amounts of a 40% CP supplement each day (M = moderate, 1.6 kg; L = low, 1.1 kg; or VL = very low, 0.5 kg; as-fed basis). After 110 d of treatment, all cows grazed dormant native grass pasture and received 1.6 kg/d of a 40% CP supplement. At 68, 109, and 123 d of treatment, cows were gathered, and plasma samples were collected by tail venipuncture (fed sample). After 18 h without feed and water, a second plasma sample was collected (fasted sample). At 109 d of treatment, BCS was greatest (P < 0.05) for H cows, similar for M and L cows, and least for VL cows. Concentrations of insulin and leptin were greater (P < 0.05) for H cows than for M and VL cows at 68 and 109 d, but similar for all groups at 123 d. Thyroxine in plasma was greatest (P < 0.05) for H cows at 68 d and similar for cows on all treatments at 123 d. Concentrations of IGF-I, insulin, and leptin in fed and fasted cows were positively correlated with BCS at 109 d. Body condition was predictive of concentrations of IGF-I, insulin, and leptin when cows had different nutrient intakes, but BCS accounted for less than 12% of the variation in plasma concentrations of IGF-I, insulin, and leptin when nutrient intake was the same for all cows. We conclude that amount of nutrient intake has a greater influence than body energy reserves on IGF-I, insulin, and leptin concentrations in the plasma of gestating beef cows.     

Relationship between serum insulin-like growth factor-I and genotype during the postpartum interval in beef cows

The objective of this study was to determine the effect of genotype and week postpartum on serum concentrations of IGF-I, body condition score (BCS), BW, and ovarian function in beef cows. Cows from the following genotypes were utilized in two consecutive years: Angus (A x A; n = 9), Brahman (B x B; n = 10), Charolais (C x C; n = 12), Angus x Brahman (A x B; n = 22), Brahman x Charolais (B x C; n = 19) and Angus x Charolais (A x C; n = 24). Serum concentrations of IGF-I, BCS, and BW were determined between wk 2 and 9 postpartum. Rectal ultrasound was used to determine days postpartum to first medium (6 to 9 mm) and first large (> or = 10 mm) follicle. Averaged across genotype, BCS decreased (P < 0.05) from 5.0 +/- 0.1 on wk 3 to 4.8 +/- 0.1 on wk 6 postpartum, and BW decreased (P < 0.05) between wk 2 and 3 and again between wk 4 and 9 postpartum. Averaged over year and week postpartum, serum IGF-I concentrations were greatest (P < 0.05) in B x B cows (46 +/- 5 ng/mL) compared with all other genotypes; lowest in A x A (12 +/- 4 ng/mL), C x C (13 +/- 4 ng/mL), and A x C cows (18 +/- 3 ng/mL); and intermediate (P < 0.05) in A x B (28 +/- 3 ng/mL) and B x C (26 +/- 3 ng/mL) cows compared with all other genotypes. Serum IGF-I concentrations did not change (P > 0.10) with week postpartum in C x C, A x A, and A x C cows, but increased (P < 0.05) between wk 2 and 7 postpartum in B x C, A x B, and B x B cows. Average interval to first medium (16 +/- 2 d) and first large (35 +/- 2 d) follicle did not differ (P > 0.10) among genotypes. Serum IGF-I concentrations correlated with BCS (r = 0.53 to 0.72, P < 0.001) but not with days to first large follicle (r = -0.19 to -0.22, P > 0.10). Averaged across genotypes, cows that lost BCS postpartum had lower (P < 0.01) serum IGF-I concentrations. Cows that calved with adequate BCS (i.e., > or = 5) had greater (P < 0.01) serum IGF-I concentrations postpartum than cows that calved with inadequate BCS (i.e., < 5) but days to first large and medium follicle did not differ (P > 0.10). In conclusion, concentrations of IGF-I in serum differed among genotypes and were associated with BCS but not days to first large or medium follicle in postpartum beef cows.     

Anovulation in postpartum suckled beef cows. I. Associations among size and numbers of ovarian follicles, uterine involution, and hormones in serum and follicular fluid

Changes in sizes and numbers of ovarian antral follicles, uterine size and weight, serum hormones, and frequency and duration of suckling were examined during the postpartum anovulatory period in primiparous, suckled beef cows. Twenty-one anovulatory, suckled cows (n = 4 to 6/d) were slaughtered on d 7, 14, 28 and 42 to 56 after parturition. In addition, a total of 11 postpartum cows that had begun cyclic activity were slaughtered on d 28, 42 or 56. Blood was collected at 10-min intervals for 6 h 1 d before slaughter for measurement of prolactin, cortisol and progesterone in serum. Numbers of medium (4.0 to 7.9 mm) follicles increased fourfold (P less than .05) between d 7 and 42 to 56 in anovulatory cows, whereas numbers of small (1.0 to 3.9 mm) and large (greater than or equal to 8.0 mm) follicles did not change (P greater than .10). Uterine involution was complete by d 28. In anovulatory cows, a higher (P less than .05) proportion of largest (but not second-largest) follicles was opposite the ovary containing the corpus albicans from pregnancy (CAP). In addition, 90% of these largest follicles opposite the CAP had concentrations of estradiol greater than progesterone. In cyclic cows, however, first ovulations occurred with equal frequency on either ovary. Concentrations of prolactin or cortisol in serum or duration of suckling were not associated with changes in uterine or ovarian measurements. In conclusion, growth and function of the largest (but not second-largest) follicle were reduced when located on the ovary containing the CAP.(ABSTRACT TRUNCATED AT 250 WORDS)     

Development of a culture system for bovine granulosa cells: effects of growth hormone, estradiol, and gonadotropins on cell proliferation, steroidogenesis, and protein synthesis

The objectives of the present studies were 1) to develop a culture system that has the positive effect of serum on granulosa cell attachment and allows subsequent expression of hormonal effects in serum-free medium and 2) to determine the effect of insulin, epidermal growth factor (EGF), estradiol (E2), and growth hormone (GH) on growth, steroidogenesis, and(or) protein synthesis of bovine granulosa cells. Cells from small (1 to 5 mm) and large (greater than 8 mm) follicles were collected from cattle and cultured for either 4 or 6 d. When cells from small follicles were cultured, insulin (5 micrograms/ml) increased (P less than .05) cell numbers (cells x 10(5)/well) severalfold compared with controls. Alone, EGF (10 ng/ml), FSH (200 ng/ml), LH (200 ng/ml), E2 (2 micrograms/ml), or GH (0 to 1,000 ng/ml) had no effect on cell numbers. However, when included with insulin, 30, 100, and 300 ng/ml of GH increased (P less than .05) granulosa cell numbers on d 4 of culture. Insulin alone increased (P less than .05) progesterone production (ng.10(5) cells-1.24 h-1) by severalfold on d 4, but EGF, FSH, LH, or GH alone had no effect and E2 inhibited progesterone production. In the presence of insulin, FSH and GH (100 ng/ml) increased (P less than .05) progesterone production on d 4 of culture, whereas EGF (10 ng/ml) elicited a decrease (P less than .05) in production. In cells from both sizes of follicles, GH (300 ng/ml) increased synthesis of cellular proteins (greater than 10 kDa). In cells from only large follicles, LH (200 ng/ml) decreased synthesis and secretion of proteins (greater than or equal to 3.5 kDa). These results support the hypothesis that GH may have direct effects on bovine ovarian function.     

Buffalo (Bubalus bubali) Late Embryo and Foetus Development: A Morphological Analysis

Many researches describe the embryonic developmental features in domestic animals; however, in farm animals, they are scarce. Most farm animal studies are related to assisted reproduction and embryos transfer techniques. But, morphological features and size measure to estimate the age gestation are rarely reported in literature. Thus, in this study, we described the developmental changes in the bubaline (Bubalus bubali) concepts from 21 to 60 days of gestation. Our results revealed that buffalo embryos similar morphological characteristics similar to other mammalian species. Also, similarities between bovine and bubaline persist; except on foetal stages when buffalos have a faster development than bovine. Therefore, buffalo's gestation period exhibits some varieties and accurate embryo age is more difficult. Yet, when we use a combination of the crown–rump, macroscopic analysis and alizarin red, it is possible to describe better the whole embryogenesis stages of the buffalo and which can contribute for future reproduction researches and applications in veterinary practice.     

Effects of feeding yeast and propionibacteria to dairy cows on milk yield and components, and reproduction*

To determine the effect of supplemental feeding of Diamond V-XP yeast (XPY) alone or in combination with propionibacteria strain P169 on milk production, milk components, body weight, days to first and second ovulation, plasma insulin, and plasma and milk glucose, 31 primiparous and multiparous (MP) Holstein cows were fed one of three dietary treatments between 2 weeks prepartum to 30 weeks postpartum: (i) control (n = 10), fed a corn silage-based total mixed ration (TMR); (ii) XPY (n = 11), fed control TMR plus XPY (at 56 g/head/day); and (iii) P169+XPY (n = 10), received control TMR plus XPY plus P169 (at 6 x 10(11) cfu/head/day). After parturition, daily milk weights were recorded, and milk samples were collected twice weekly for milk component analyses. Daily uncorrected milk, solids-corrected milk, and 4% fat-corrected milk production for MP cows fed P169+XPY was 9-16% greater than control MP cows, but these increases were only evident during mid lactation (9-30 weeks). The percentage of milk fat was 8-18% greater in control than XPY and P169+XPY groups. Milk lactose percentage in MP cows fed P169+XPY was 3-5% greater than in control and XPY MP cows. Primiparous and MP cows fed P169+XPY had 28-32% greater milk glucose levels than control and XPY-fed cows. Diurnal plasma glucose concentration was not affected by diet in MP cows. Plasma insulin levels in MP cows fed P169+XPY were 30-34% greater than in other groups of MP cows. Milk glucose and plasma insulin responses to P169+XPY feeding suggest that P169+XPY might have enhanced gluconeogenesis and increased glucose uptake by the mammary gland in Holstein cows. Thus, a combined feed supplement of P169 and XPY may hold potential as a natural feed alternative to hormones and antibiotics to enhance lactational performance.