Effect of Ovarian Status on Reproductive Performance in Dairy Heifers Inseminated with Sexed Semen

Background: The intensive reproductive management in the dairy farms requires inclusion of a large group of replacement heifers in the breeding program for a shorter period. In this aspect, a creation of effective estrus synchronization protocols with timed artificial insemination (TAI) by sexed semen and optimization of the current ones have a crucial role for obtaining high pregnancy rate. These protocols are beneficial, because they led to reduced interval to first AI, lack of need for estrus detection, and allow obtaining a large group of female calves for on time. Because of limited fertilizing potential of sexed spermatozoa this type of semen is applied mainly for heifers, as fertility is higher compared to lactating cows. The main objective of the present study was to investigate the effect of ovarian status on the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Materials, Methods & Results: Forty-eight Holstein healthy heifers separated in 2 groups were subjected to PGF2α-GnRH pre-treatment and Ovsynch or PRID-5-day estrus synchronization protocol and timed artificial insemination (TAI) with sex-sorted semen, staring 6 days after end of the hormonal pre-treatment. The ovarian status (presence of follicles with or not a corpus luteum) of the heifers at the begin of the treatment, on day of TAI and pregnancy rate in different groups were determined and compared. Additionally, the ovarian status at the begin of the treatment, ovarian status and size of preovulatory follicle (PF) on day of TAI and total values for both groups according to reproductive performance (pregnant or non-pregnant) were also analyzed. On day of TAI the animals with PF and a lack of corpus luteum (CL) in both groups were more that those with PF and CL (39.3% and 30% vs. 60.7 % and 70%), with significant (P < 0.05) difference in PRID-5-day group. The pregnancy rate tended to be higher in PRID-5 day than Ovsynch treatment (65% vs. 35.7%). A higher percentage (100% and 67.9%) of the pregnant animals in both treatments had not CL on day of TAI, and the size of the PF (1.58 ± 0.12 cm and 1.64 ± 0.13 cm) was increased (P < 0.05). Similar effects of the ovarian status on reproductive performance were obtained after a comparison of the total values between pregnant and non-pregnant animals. The percentage of heifers with observed PF without CL on day of TAI was significantly greater (P < 0.05) in pregnant compared to non-pregnant group (91.3% vs. 40%). The opposite dependence was determined for the parameter presence of follicles and corpus luteum (8.7% vs. 60%; P < 0.05).  Furthermore, the size of the PF measured immediately before TAI was increased in animals became pregnant (1.60 ± 0.12 cm vs. 1.34 ± 0.17 cm; P < 0.05). Discussion: The analysis of the obtained results showed that the ovarian status on day of TAI affects the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Hormonal pre-treatment with onset of PRID-5-day protocol 6 days later and TAI with sex-sorted semen ensure acceptable pregnancy rate. The greater preovulatory follicle and a lack of corpus luteum before insemination provide significantly (P < 0.05) more pregnant animals, compared to the cases when CL is presented. Ultrasound determination of the ovarian function before insemination can be used in selection of heifers for TAI with sex-sorted semen.                   Keywords: heifers; estrus synchronization; sexed semen, pregnancy

Pregnancy Rates Associated with Oxidative Stress after Estrus Synchronization of Bulgarian Murrah Buffaloes in Breeding and Non-Breeding Season

Background: The current study aims to measure the effect of oxidative stress on the pregnancy rates of Bulgarian Murrah buffaloes during the breeding and non-breeding season. Methods: The study group consisted of 24 mature buffaloes more than 40 days after parturition. The following parameters were measured: Reactive Oxygen Species (ROS) products, Ascorbate radicals, Malondialdehyde (MDA), Nitric Oxide (NO), Super Oxide Dismutase (SOD), Glutathione peroxidase (GSH-Px), Protein Carbonyl Content (PPC), and total Nitric oxide. The Presynch/Ovsynch protocol was used for estrus synchronization. Results: A statistically significant increase in ROS products were measured in blood serum during the breeding season compared with the non-breeding season. The highest levels measured were in non-pregnant buffaloes during the breeding season. High levels of oxidative stress were registered due to low SOD activity in buffaloes during the breeding season compared to SOD activity during the non-breeding season. The highest SOD activity was observed in non-pregnant buffaloes during the summer season. The lowest GSH-Px levels were observed in non-pregnant buffaloes during both study periods. During the breeding season, concentrations of total NO and PPC were elevated. Conclusion: Comparing the obtained results for oxidative stress and antioxidant activity concerning pregnancy rate depending on the season showed that pregnancy in buffaloes during the breeding season was realized at higher values of NO and SOD. Increased oxidative stress was observed, resulting in a statistically significant increase in serum ROS products, as well as decreased SOD activity in buffaloes during the breeding season

Effect of Ovarian Status on Reproductive Performance in Dairy Heifers Inseminated with Sexed Semen

Background: The intensive reproductive management in the dairy farms requires inclusion of a large group of replacement heifers in the breeding program for a shorter period. In this aspect, a creation of effective estrus synchronization protocols with timed artificial insemination (TAI) by sexed semen and optimization of the current ones have a crucial role for obtaining high pregnancy rate. These protocols are beneficial, because they led to reduced interval to first AI, lack of need for estrus detection, and allow obtaining a large group of female calves for on time. Because of limited fertilizing potential of sexed spermatozoa this type of semen is applied mainly for heifers, as fertility is higher compared to lactating cows. The main objective of the present study was to investigate the effect of ovarian status on the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Materials, Methods & Results: Forty-eight Holstein healthy heifers separated in 2 groups were subjected to PGF2α-GnRH pre-treatment and Ovsynch or PRID-5-day estrus synchronization protocol and timed artificial insemination (TAI) with sex-sorted semen, staring 6 days after end of the hormonal pre-treatment. The ovarian status (presence of follicles with or not a corpus luteum) of the heifers at the begin of the treatment, on day of TAI and pregnancy rate in different groups were determined and compared. Additionally, the ovarian status at the begin of the treatment, ovarian status and size of preovulatory follicle (PF) on day of TAI and total values for both groups according to reproductive performance (pregnant or non-pregnant) were also analyzed. On day of TAI the animals with PF and a lack of corpus luteum (CL) in both groups were more that those with PF and CL (39.3% and 30% vs. 60.7 % and 70%), with significant (P < 0.05) difference in PRID-5-day group. The pregnancy rate tended to be higher in PRID-5 day than Ovsynch treatment (65% vs. 35.7%). A higher percentage (100% and 67.9%) of the pregnant animals in both treatments had not CL on day of TAI, and the size of the PF (1.58 ± 0.12 cm and 1.64 ± 0.13 cm) was increased (P < 0.05). Similar effects of the ovarian status on reproductive performance were obtained after a comparison of the total values between pregnant and non-pregnant animals. The percentage of heifers with observed PF without CL on day of TAI was significantly greater (P < 0.05) in pregnant compared to non-pregnant group (91.3% vs. 40%). The opposite dependence was determined for the parameter presence of follicles and corpus luteum (8.7% vs. 60%; P < 0.05).  Furthermore, the size of the PF measured immediately before TAI was increased in animals became pregnant (1.60 ± 0.12 cm vs. 1.34 ± 0.17 cm; P < 0.05). Discussion: The analysis of the obtained results showed that the ovarian status on day of TAI affects the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Hormonal pre-treatment with onset of PRID-5-day protocol 6 days later and TAI with sex-sorted semen ensure acceptable pregnancy rate. The greater preovulatory follicle and a lack of corpus luteum before insemination provide significantly (P < 0.05) more pregnant animals, compared to the cases when CL is presented. Ultrasound determination of the ovarian function before insemination can be used in selection of heifers for TAI with sex-sorted semen.                   Keywords: heifers; estrus synchronization; sexed semen, pregnancy

Oxytetracycline Pharmacokinetics After Intramuscular Administration in Cows with Clinical Metritis Associated with Trueperella Pyogenes Infection

Systemic therapy with oxytetracycline is often used for treatment of clinical metritis although data about its penetration into the uterus and uterine secretion are lacking. Uterine secretions and milk from six cows with clinical metritis were collected for microbiological assay. The animals were treated intramuscularly with long-acting oxytetracycline (20 mg/kg) and samples of plasma, milk and uterine secretions were collected for determination of the antibiotic concentrations by HPLC-PDA analysis. Pharmacokinetics of the antibiotic and in silico prediction of its penetration into the uterus were described. Trueperella pyogenes with MIC values of 16–64 µg mL−1 was isolated (n of cows = 4) from uterine secretions. Oxytetracycline showed fast absorption and penetration in the uterine secretions and milk. No change of withdrawal time for milk was necessitated in cows with clinical metritis. Maximum levels in uterine secretions and predicted concentrations of oxytetracycline in the uterus were lower than MIC values. Systemic administration of long-acting oxytetracycline did not guarantee clinical cure and was not a suitable choice for treatment of clinical metritis associated with Trueperella pyogenes. The appropriate approach to antibiotic treatment of uterine infections of cows requires knowledge on penetration of the antibiotics at the site of infection and sensitivity of pathogens

Resumption of Cyclic Ovarian Activity by Herbal Preparation AyuFertin in Bulgarian Murrah Buffaloes at Early Postpartum

This study evaluates the phytochemical composition and ability of herbal preparation AyuFertin, supplemented shortly after calving, to resume the cyclic ovarian activity in postpartum Bulgarian Murrah buffaloes. In total, 13 healthy Bulgarian Murrah buffaloes at the age of 4–10 years, calved in March–April 2019 were involved in the experiment. Seven experimental buffaloes were treated individually per os with AyuFertin according to producer instruction. All animals were subjected to regular ultrasound examination of ovaries. GC-MS analysis of fatty acids composition and HPLC-DAD analysis of carotenoid and tocopherol content in AyuFertin were conducted. The determination of estrus occurrence and natural mating were done by a fertile bull. The development of the large follicles (12.5–14.3 mm) in 85.7% of treated animals was observed on day 24 postpartum compared to 0% in controls. Clinical signs of estrus were recorded in 71.4% of the animals, followed by an 80% pregnancy rate versus 16% in controls within 70 days after calving. In conclusion, the supplementation of the bioactive herbal product AyuFertin from day 21 postpartum can stimulate the follicular growth in the buffalo’s ovary, which leads to the development of follicles with ovulatory capacity, followed by successful ovulation. The above-mentioned treatment resulted in a better pregnancy rate in the treated multiparous compared to the control buffaloes bred by natural service