C. Murcia

Effect of blood metabolites, body condition and pasture management on milk yield and postpartum intervals in dual-purpose cattle farms in the tropics of the state of Veracruz, Mexico.

Research was conducted on typical smallholder farms with dual-purpose cattle (DPC) (Bos indicus x B. taurus) in the coastal north-central area of Veracruz, Mexico. The study was divided into two phases. The aim of the first phase was to investigate the effect of blood metabolities, body condition and pasture management on milk yield and postpartum intervals, in order to investigate if the former are suitable indicators of the reproductive and nutritional status of DPC. One hundred and sixty-five calvings of crossbred cows were recorded from January 1992 to November 1994 on 12 small farms. Milk samples were collected twice a week for progesterone analysis. Blood samples and BCS were taken once a month. However, in Phase II emphasis was placed on the effect of pasture management upon reproductive and productive performance of DPC. Records of four farms were obtained from June 1995 to November 1996. Stocking rates were 0.40, 0.87, 0.35 and 1.5 cows/ha for farms A, B, C and D, respectively. Farms A and C used a slow rotation while B and D used a rapid rotation. In Phase I, the changes in BCS during the last month of pregnancy and first month postpartum did not correlate (p > 0.05) with milk yield or reproductive performance. Blood the metabolite profiles were not consistently related to productive or reproductive variables. The effect of farm and season was significant (p < 0.05) on most of the response variables and low productivity on overstocked farms lead to the conclusion that the low reproductive performance of DPC was linked to poor pasture management. During Phase II, farms A (FA) and D (FD) produce more milk than the others. Days to first service, days open, and calving interval were similar for farms B (FB) and C (FC), highest for Farm A, and lowest for Farm D. The forage availability mean was above the critical range of 6-8 kg of dry matter per 100 kg of liveweight (kg DM/100 kg LW) in all farms (range from 6.1 +/- 5.0 to 21.1 +/- 11.2 kg DM/100 kg LW). Farm D had the highest stocking rate (1.5 cows/ha), a rapid rotation (10 paddocks), a good forage availability (7.1 +/- 3.9 kg DM/100 kg LW) with a good quality for a tropical pasture (11.6 +/- 2.4% crude protein), and an economic energy supplementation. These results suggest this type of management could be more widely employed to improve the productivity of DPC on smallholder farms in the Mexican tropics.

Ovine serum and pituitary isoforms of luteinising hormone during the luteal phase

The relative abundance of the different isoforms of pituitary and circulating luteinising hormone (LH) in ewes, at different times after the administration of gonadotrophin-releasing hormone (GnRH), during the luteal phase of the oestrous cycle was investigated. Sixteen ewes on Day 9 of their cycle were divided into four groups (n = 4). The control group (T0) received saline solution; the remaining animals received 100 microg GnRH (i.m.) 30, 90 or 180 min (T30, T90 and T180, respectively) before serum and pituitary gland collection. Luteinising hormone polymorphism was analysed by chromatofocusing (pH 10.5-3.5). The LH eluted from each chromatofocusing was grouped on the basis of the following three criteria: (1) according to the pH of elution (pH > or = 10-3.5); (2) as either a basic (pH > or = 7.5), neutral (pH 7.4-6.5) and acidic (pH < or = 6.4) elution of LH of serum and hypophyseal origin; and (3) on the basis of distinct isoforms, of which 10 (A-J) were identifiable in hypophyseal extracts and four (A-D) were found in the serum. In general, the most abundant forms of LH in both the pituitary and serum, at all times, were basic. However, that proportion was greater in hypophyseal extracts (84 +/- 3%, 81 +/- 4%, 82 +/- 3% and 83 +/- 2% at T0, T30, T90 and T180, respectively) than in serum (51 +/- 5%, 48 +/- 10% and 54 +/- 6% at T30, T90 and T180, respectively). Neutral and acidic LH made up a larger proportion of the total LH in sera (neutral: 17 +/- 4%, 20 +/- 6% and 23 +/- 3% at T30, T90 and T180, respectively; acidic: 32 +/- 8%, 32 +/- 11% and 23 +/- 6% at T30, T90 and T180, respectively) than in the pituitary extracts (neutral: 4.0 +/- 0.7%, 10 +/- 4%, 7 +/- 2% and 5.0 +/- 0.5% at T0, T30, T90 and T180, respectively; acidic: 12 +/- 3%, 11 +/- 2%, 12 +/- 2% and 12 +/- 2% at T0, T30, T90 and T180, respectively) at all times. These data reveal that the relative composition of the LH present in the pituitary gland and the LH secreted into the circulation is different, with more neutral and acidic isoforms being secreted. The pattern of circulating LH isoforms changes between 30 and 180 min after GnRH peak induction, with a greater proportion of isoform C (eluting between pH 7.0 and 6.5) at T180 compared with T30 and T90.

Reduced response to an estrous induction program in postpartum beef cows treated with zilpaterol and gaining body weight.

In beef cows, reduced energy intake delays first ovulation postpartum and is associated with lesser insulin, IGF-I and leptin concentrations. However, the close relationship among these hormones mask their individual roles in the reinitiation of ovarian activity. A β-adrenergic receptor agonist (βAR) was used to increase body condition score (BCS) and yet reduce body fat and leptin serum concentration to determine the specific role of leptin in the postpartum ovarian activity. Beef cows (n=77) with BCS 3.1 ± 1.4 received 2 kg/day of feed containing 0 or 0.15 mg/kg of zilpaterol (a synthethic βAR), for 33 days. Estrus was induced with a progestin implant applied for 9 d and cows in estrus were bred by artificial insemination (AI). Zilpaterol administration increased (P<0.05) daily weight gain, muscle depth and BCS, with no changes in back fat depth, reducing fat to muscle ratio (P<0.05). At the time of AI, insulin (38%) and IGF-I (26%) concentrations were less in zilpaterol-treated cows (P<0.05), but leptin concentration was unaffected. Ovulation rate and animal with luteal activity after estrus induction were also reduced by 35% (P=0.05) and 56.5% (P=0.007), respectively, in zilpaterol-treated cows. Logistic regression estimates for BCS (P=0.016) and IGF-I concentration (P=0.03) were positively related with the occurrence of luteal activity. In addition, whilst back fat (P=0.009) had a positive effect on luteal activity, leptin concentration did not show a significant relationship. In conclusion, despite an increase in body weight and a positive change in BCS, the reduction in insulin and IGF-I concentrations, associated with βAR treatment, reduced the response to induction of estrus. However only IGF-I, but not leptin or insulin, significantly influenced the odds for the occurrence of luteal activity after estrous induction in cattle with poor BCS.

Effect of ovine luteinizing hormone (oLH) charge isoforms on VEGF and cAMP production.

Although an increase in VEGF expression and synthesis in association with LH has been established; it is unknown if all LH isoforms act similarly. This study evaluated the production of cAMP and VEGF among LH isoforms in two in vitro bioassays. The LH was obtained from hypophyses and the group of isoforms was isolated by chromatofocusing. cAMP production was assessed using the in vitro bioassay of HEK-293 cells and VEGF production was evaluated in granulosa cells. Immunological activity was measured with a homologous RIA. Immunoactivity and bioactivity for each isoform were compared against a standard, by estimating the IC50 and the EC50. The basic isoforms were more immunoactive than the standard. The neutral and the moderately acidic had an immunological activity similar to the standard. The acidic isoform was the least immunoreactive. cAMP production at the EC50 dose was similar among the basic isoforms, the moderately acidic and the standard; for the neutral and the acidic, the EC50 dose was higher. It was observed that compared with the control, VEGF production at the lowest LH dose was no different in the standard and each isoform. In the intermediate dose, a positive response was caused in the standard and the neutral and basic isoforms. Although the acidic isoform showed a dose-dependent response, it was not significant relative to the control. In conclusion, the basic isoform generated the greatest cAMP and VEGF production, similar to the reference standard, and the acidic the smallest.

The presence of VEGF and Notch2 during preantral-antral follicular transition in infantile rats: Anatomical evidence and its implications

Folliculogenesis is a process that depends on angiogenesis, in which VEGF and Notch signaling pathway members are involved. Although this pathway is present in preantral and antral follicular structures during the second stage of folliculogenesis, this association has not been described. Therefore, this study aimed to identify VEGF and Notch2 in ovary structures of infantile rats after induction of follicular development with a gonadotropin stimulus. In order to explore this possibility we analyzed rat ovary morphology from days 10-25 after birth; subsequently, the transition from preantral follicle to an antral stage was analyzed by the induction of follicular development with equine chorionic gonadotropin (eCG) and VEGF and Notch were identified in the rat ovary by fluorescence. The histological analysis revealed that the ovary of a 10-day-old rat has the highest percentage of preantral follicles and based on this a 10IU eCG dose promoted an increase in the number of antral follicles, as well as a decrease in the number of preantral follicles, related to which there was an increase in ovary weight and size. In addition, a higher concentration of circulating estradiol was observed, proliferation of granulosa cells in both follicle groups was stimulated, and the accumulation of VEGF in granulosa and theca cells and in the antral follicle oocyte was increased (p<0.05), whereas the presence of Notch2 was limited to mural granulosa cells, in granulosa cells that formed the cumulus oophorus and in the oocyte of both groups of follicles. The multiple correspondence analysis allowed us to support an association between VEGF and Notch2 during the transition from preantral to antral follicles in the ovary of an infantile rat.