Use of GnRH-Encapsulated Chitosan Nanoparticles as an Alternative to eCG for Induction of Estrus and Ovulation during Non-Breeding Season in Sheep.

This study is aimed at determining the reproductive performance of anestrous ewes treated with nanoencapsulated GnRH after a progesterone-based protocol for estrus induction was proposed as a way of replacing eCG. A total of sixty anestrous, multiparous, non-lactating Barki ewes were randomly allocated into three homogenous groups and subjected to a CIDR-based estrus induction protocol. The first group (eCG) received an intramuscular (i.m.) injection of 350 IU of eCG at CIDR removal. The second (LNGnRH) and third (HLNGnR) groups received either 25 µg or 50 µg of encapsulated GnRH nanoparticles by the i.m. route in the form of spherical GnRH-encapsulated chitosan-TPP nanoparticles (which were 490.8 nm and had a 13.6 mV positive charge) 48 h after CIDR removal. Follicular dynamics, estrous behavior, luteal activity, and pregnancy outcomes were evaluated. Three days after CIDR removal, the number of large follicles increased by similar amounts in the LNGnRH and eCG groups and were significantly higher in both groups than in the HNGnRH group. However, no differences were observed in the numbers and diameters of CLs among the experimental groups and, on the other hand, treatment with HNGnRH significantly increased blood serum progesterone levels compared with eCG and LNGnRH. Treatment with HNGnRH increased conception, lambing, and fecundity rates (p < 0.05), with the trend of a higher litter size (p = 0.081) compared with eCG, whereas LNGnRH resulted in intermediate values. In conclusion, a dose of 50 µg of GnRH encapsulated in chitosan-TPP nanoparticles can be used as an alternative to eCG in progesterone-based estrus induction protocols in sheep.

The Effect of P4 + eCG Estrus Induction Protocol during the Deep and the Transition Anestrous Period on the Reproductive Performance of Crossbred Dairy Goats.

Seasonal reproduction restricts the offering of goat commodities across the year. Therefore, it is crucial to improve diverse strategies to induce the reproductive response in goats during the anestrus stage. The effectiveness of a short P4 + eCG-based estrus induction protocol during both the deep anestrous (March) or the reproductive transition period (June) upon the reproductive performance of crossbred dairy goats was assessed. Adult, anestrous, 24-30-month- old dairy crossbred (Saanen-Alpine-Nubian × Criollo) goats (n = 123) from two commercial herds and 10 sexually active goat bucks were used. Before the trials, the anestrous status of goats was confirmed. Then, goats were randomly allocated into two different experiments. In Exp. 1, we tested the effect of different doses (D) of intramuscular progesterone (P4; 10 or 20 mg + eCG (100 UI)) and type of breeding (TB), natural mating (NM), or artificial insemination (AI), on two commercial goat herds (H1 & H2), in March (deep anestrous). In Exp. 2, we evaluated the effect of D (P4; 10 or 20 mg + 100 UI eCG) in goats subjected to NM, and either during deep anestrous (March; M) or transitional anestrous (June; J), in two commercial herds. After breeding, conception and pregnancy were diagnosed with ultrasound scanning. The response variables were estrus induction (EI; %), estrus latency (EL; h), ovulation (OVP; %), ovulation rate (OR; units), fertility (FERT; %), and pregnancy (PREG; %). No differences (p > 0.05) in live weight (LW) and body condition score (BCS) occurred between herds in both trials. In Exp. 1, EI, EL, OVP, OR, FERT, and PREG were affected (p < 0.05) by the H-TB-D interaction, whereas in H1 + P4-20 combination had the highest (p < 0.05) EI, EL, and OVP values. Irrespective of TB, H1 had the largest (p < 0.05) OR, independently of TB or D. Also, the lowest (p < 0.05) OVP occurred in the AI + P4-10 group, while the AI had the lowest (p < 0.05) FERT, irrespective of D. FERT and PREG were two-fold higher (p < 0.05) in NM compared with AI. In Exp. 2, EI, EL, OVP, OR, FERT, and PREG were affected (p < 0.05) by the H-M-D interaction. In general, H2 + P4-10 had the lowest (p < 0.05) reproductive outcomes in March, whereas H1 had the largest (p < 0.05) values in either month. No differences (p > 0.05) between P4 doses occurred for EI, OVP, OR, FERT, and PREG. Yet, the largest (p < 0.05) EL occurred with P4-20 in June. No correlations (p > 0.05) occurred between LW and all the reproductive variables. BCS was positively correlated (p < 0.05) with EI (0.34), OVP (0.44), OR (0.58), and PREG (0.20). Also, positive correlations (p < 0.05) occurred between EI with EL (0.83), OVP (0.80), OR (0.64), and PREG (0.56); EL with OVP (0.58), OR (0.44), and PREG (0.42); OVP with OR (0.79) and PREG (0.70), as well as OR and PREG (0.63). Results of these studies confirm a multidimensional response regarding the effectiveness of P4 + eCG for estrus induction in goats mainly modulated by a specific time within the anestrous season, or even by specific management or a particular environment at the herd level (H1), although quite remarkably independent of the animal's LW or BCS at herd level. Moreover, the best reproductive outcomes occurred with NM in June. The most reproductive variables were similar using either 10 or 20 mg P4 + 100 IU eCG, giving the possibility to lessen the scale in the use of exogenous hormones while obtaining acceptable out of season reproductive response.

Temporal concentrations of cortisol and LH in virgin ewes acutely exposed to rams during the transition into the breeding season.

The objectives of this study were to determine if exposing seasonally anovular ewes to rams would alter patterns of cortisol concentrations, and if these changes are associated with changes in characteristics of LH concentrations. Seasonally anestrous ewes were assigned to be exposed to rams (RE; n=11) or wethers (NE; n=12). Blood samples were collected at 15-min intervals beginning 120 min before introduction of males (time=0 min), and continued for 360 min after male exposure. Characteristics of cortisol and LH concentrations included: mean and baseline concentrations, pulse amplitude, duration, frequency, and time to first pulse. Mean and baseline cortisol concentrations, and cortisol pulse amplitude, frequency, and time to first pulse after male exposure did not differ between RE and NE ewes. Cortisol pulse duration was longer (P<0.05) in RE ewes than in NE ewes. Mean LH and LH pulse amplitude, duration, and time to first pulse after male exposure did not differ between RE and NE ewes. Baseline LH concentrations and LH pulse frequency were greater (P<0.05) in RE than in NE ewes. In RE ewes, but not NE ewes, LH pulse frequency tended to increase (P=0.06) as pulse frequency of cortisol decreased. In conclusion, exposing ewes to mature rams during the transition into the breeding season increased LH pulse frequency which hastened ovulatory activity. However, the results do not support the hypothesis that changes in cortisol concentrations plays a significant role in the 'ram effect'.

Importance of intense male sexual behavior for inducing the preovulatory LH surge and ovulation in seasonally anovulatory female goats.

The present study was carried out to determine whether the presence of photostimulated sedated male goats could stimulate the LH preovulatory surge and ovulation in seasonal anestrous goats. Sexually experienced male goats were treated with artificial long days (16 hours light per day) from 1 November to 15 January to stimulate their sexual activity in March and April, corresponding to the natural sexual rest. A female group of goats (n=20) was exposed to non-sedated males who displayed an intense sexual behavior and provided strong odor (non-sedated group). Another female group of goats (n=20) was exposed to the photo-stimulated male goats, but these males were sedated with Xylazine 2% to prevent the expression of sexual behavior (sedated group). The sedated males also provided a strong odor. Females of both groups had full physical and visual contact with non-sedated or sedated males. In both groups, the males remained with females during 4 days. The LH preovulatory surge of 10 female goats per group was measured by determination of LH plasma concentrations in samples taken every 3 hours. In addition, in all goats, (n=20 by group), ovulation was determined by measuring plasma concentrations of progesterone. The proportion of female goats showing a preovulatory LH surge was higher in goats exposed to non-sedated (10/10) than in those exposed to sedated bucks (0/10; P<0.0001). Similarly, most of does in contact with non-sedated males ovulated (19/20), but none of those in contact with sedated males did so (0/20; P<0.0001). We conclude that the expression of an intense sexual behavior by male goats is necessary to induce LH preovulatory surge and ovulation in seasonally anovulatory goats.