We investigated the effects of gonadotropin releasing hormone (GnRH) agonist on expressions of GnRH receptor (GnRHR), follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) proteins in the ovaries and follicular development in the ewes. Forty-two pre-pubertal ewes were assigned to experimental groups 1 to 5 (EG-I to EG-V) and control group (CG). Ewes in EG-I, EG-II and EG-III were subcutaneously injected with 200, 300 or 400 μg alarelin antigens twice (on days 0 and 14), respectively. Ewes in EG-IV and EG-V were subcutaneously injected with 200 μg and 300 μg alarelin antigen four times (on days 0, 7, 14 and 21). Ewes in CG were subcutaneously injected with a solvent twice (on days 0 and 14). Serum concentrations of GnRH antibody in the EGs increased and were higher than (P<0.05) that of CG from day 14 to day 60. GnRH antibody concentrations in EG-IV and EG-V were higher than that in EG-I, EG-II and EG-III from days 35 to 45. Expressions of GnRHR protein in EG-IV and EG-V were lower than that in CG (P<0. 01). Expressions of FSHR and LHR proteins in EGs increased. Levels of FSHR and LHR proteins in EG-IV and EG-V (P<0.05) were higher than CG. Ovarian weights in EGs increased. Values of follicle vertical diameter, follicle transverse diameter, follicle wall thickness, follicle externatheca thickness and follicle internatheca thickness in EG-III and EG-V were greater than other groups. Primordial follicles and primary follicles developed quickly in alarelin-immunized animals. Secondary follicles and mature follicles became more abundant. Mitochondria, mitochondrial cristaes and cortical granules increased. Serum FSH concentrations of EGs remained higher than that in CG from days 28 to 70 (P<0.05). Alarelin immunization stimulated GnRH antibody production, suppressed expression of GnRHR protein, enhanced expressions of FSHR and LHR proteins in ovaries, promoted FSH secretion and thereby accelerated the development of ovaries and follicles in ewes.

Peptides derived from food proteins have exhibited significant antihypertensive effects without side effects. In this study, the cottonseed protein was hydrolyzed by papain for preparing peptide with angiotensin I converting enzyme (ACE) inhibitory activity. The influence of hydrolysis temperature (33°C, 40°C, and 47°C), pH (6.5, 7.0, and 7.5), and enzyme to substrate (E/S) ratio (0.9%, 1.2%, and 1.5%) on the degree of hydrolysis (DH) and ACE-inhibitory activity were analyzed. The hydrolysis conditions were further optimized by central composite design (CCD) and response surface methodology (RSM). The DH of cottonseed protein and ACE inhibitory rate of hydrolysates reached 25.7% and 88.2%, respectively, under the optimal hydrolysis conditions (hydrolysis temperature 39°C, pH 7.5, and E/S ratio 1.04%). We further separated the cottonseed protein hydrolysates (CPH) using a combined strategy of ultrafiltration membrane bioreactor system, sephadex G-25 gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC). An ACE inhibitory peptide, named as FII-2-P, with ACE inhibitory IC 50 value of 46.7 μg/mL, was obtained. MALDI-TOF-TOF mass spectrometry analysis revealed that the molecular weight of FII-2-P was 763.4 Da and its amino acids sequence was Phe-Pro-Ala-Ile-Gly-Met-Lys. These results demonstrated that the cottonseed protein is a potential source of ACE inhibitory ingredients to be used in the development of functional foods.

This study aims to investigate the estrus efficacy of FIS (fluorogestone intravaginal sponges)/eCG (equine chorionic gonadotropin) and cloprostenol treatments to establish the optimum protocols for estrus synchronization in ewes. Forty-two ewes were assigned into groups A (FIS/eCG group) and B (CLO (cloprostenol) group). Each group was randomly divided into three subgroups (n = 7) according to the dose of eCG (ECG1, ECG2 and ECG3) or cloprostenol (CLO1, CLO2 and CLO3). The estrus rates, lambing rates and average litter sizes were accounted. Serum concentrations of progesterone (P4), folliclestimulating hormone (FSH) and luteinizing hormone were determined. Selected optimum schemes were employed in 636 ewes in four sheep farms. Estrus rates of groups A and B were 85.72% and 57.14%. Estrus onset times (EOTs) were 53.91 ± 12.26 and 46.41 ± 4.65 h for groups A and B. EOT of ECG1 was longer than that of CLO3 (P < .05). Lamb numbers, lambing rate and litter size of group B were lower than those of group A. The maximum and minimum lambing rates were calculated in ECG3 and CLO1 (157 vs. 57). On day 11, higher P4 values (P < .05) were determined of ECG3 and CLO1 compared to CLO2 and CLO3 subgroups. FSH concentration of ECG1 was significantly higher than that in group B (P < .05). Application results showed that estrus rate, pregnancy rate and lambing rates of ECG3 and CLO3 protocols were 95.97% vs. 70.91%, 91.37% vs. 88.89% and 149.87% vs 132.69% (P < .05), respectively. FIS/eCG and cloprostenol improved estrus synchronization. Injection of 400 IU eCG after FIS withdrawal was an optimum protocol for estrus synchronization of ewes. ARTICLE HISTORY

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