Cryopreservation poses significant challenges to the preservation of sperm integrity and function, particularly in small ruminants where cryodamage is pronounced. This review explores the molecular mechanisms underlying sperm cryodamage and strategies for improving cryopreservation outcomes, with a focus on the role of antioxidants. Cryopreservation-induced alterations in proteins and RNA transcripts critical for sperm function, including motility, capacitation, fertilization, and embryo development, are discussed. Proteomic, transcriptomic, and epigenomic advancements have provided valuable insights into these mechanisms, offering potential biomarkers for predicting sperm freezability and enhancing cryopreservation strategies. Combining technologies such as mass spectrometry and flow cytometry allows for a comprehensive understanding of molecular and cellular changes induced by the freezing–thawing process. However, challenges remain in optimizing cryoprotectant formulations and antioxidant supplementation to improve post-thaw sperm fertility. Further research is needed to explore a wider range of novel cryoprotectants, antioxidants, and proteins for cryopreservation media, as well as to validate their efficacy in enhancing sperm viability and function. Additionally, investigations into the effects of cryopreservation on RNA transcripts and epigenetic factors in small ruminant species are warranted to advance our understanding of sperm preservation. Overall, this review highlights the importance of antioxidants in mitigating cryodamage and underscores the need for continued research to refine cryopreservation protocols and improve reproductive outcomes in small ruminants.

This review aimed to bring a comprehensive analysis of key clinical strategies for enhancing reproductive efficiency in buffaloes, a species that exhibit low reproductive performance under conventional reproductive management compared to that exhibited by cattle. It considers key ART techniques including estrus synchronization for artificial insemination, and ovulation induction, highlighting their role in improving fertility and overall herd productivity. However, it also addresses common postpartum inflammatory and functional reproductive disorders, discussing their diagnosis and treatment protocols, stressing their impact on the overall reproductive outcome in buffalo farming.

Background: Seasonal anestrus in ewes reduces reproductive efficiency and hinders productivity. Exogenous hormones can be used to facilitate the induction and synchronization of estrus in the anestrus period. The aim of this study was to evaluate the pregnancy rate in ewes after the treatment with different hormones and fixed time artificial insemination. Methods: The ewes were randomly divided in 4 (n=4) groups: Melatonin group (Group 1), Melatonin and Medroxyprogesterone group (Group 2), Medroxyprogesterone group (Group 3) and control group (Group 4). Pregnancy rate, estrus induction rate and litter size were determined and compared. Result: The estrus induction rate in our study was 100% for the first 3 groups and 6.25% for the 4th group. The pregnancy rate was between 4.16% (group 4) and 95.83% (group 2). For the others groups the values were 89.58% for the first group and 91.67%, for the 3rd group. The prolificacy rate was between 1 (control group) and 1.71 (2nd group), with intermediate values for the other 2 groups: 1.21 for 1st group and 1.27 for the 3rd group.

Introduction: This study investigated the association between postpartum subclinical hypocalcemia (SCHC) and reproductive and economic performance in dairy cows. SCHC is often overlooked yet may contribute significantly to reduced productivity and profitability in dairy herds.Methods: A total of 312 cows, Holstein Friesian and Romanian Spotted breeds, from three commercial farms in Eastern Europe were monitored during the first 60 days postpartum. Blood calcium levels were measured to classify cows into SCHC and normocalcemic groups.Results: The incidence of SCHC was 42.9%, with multiparous cows more frequently affected. Cows with SCHC had longer service periods, extended calving intervals, and required more artificial insemination attempts than normocalcemic cows. Although milk yield did not differ significantly between groups, SCHC cows exhibited moderately higher mastitis incidence and somatic cell counts, especially within the first 30 days postpartum. These factors contributed to increased indirect costs. Economic analysis indicated that SCHC cows incurred approximately 54 EUR more in additional costs per animal compared to normocalcemic cows.Discussion: SCHC is a hidden contributor to economic losses in dairy operations, primarily due to its negative impact on reproductive performance and udder health. These findings underscore the importance of early detection and preventive strategies to mitigate the effects of SCHC and improve herd productivity during the transition period.

This study investigated the reproductive and economic costs of four estrus synchronization protocols in multiparous Holstein dairy cows: Ovsynch with one or two artificial inseminations (AIs), and Double Ovsynch with one or two AIs. A total of 216 clinically healthy cows were randomly assigned to one of the four treatment groups (n = 54 each). Pregnancy diagnosis was performed by transrectal ultrasonography at 33 days post insemination. Economic calculations included the cost of hormonal treatments, insemination, and pregnancy diagnosis, reflecting market prices in large scale Romanian dairy operations. The highest pregnancy rate (64.8%) and lowest cost per confirmed pregnancy (EUR 89.51) were achieved in the group treated with Double Ovsynch followed by a single AI. The standard Ovsynch protocol with one AI yielded the lowest conception rate (42.6%), indicating limited effectiveness under fixed-time AI conditions. These findings suggest that Double Ovsynch with a single insemination is the most effective and economically viable protocol for reproductive management in intensively managed dairy herds.

Backgroung: Long term storage of spermatozoa is one of the most desirable tools of assisted reproductive technologies. The purpose of this research was to evaluate the effect of two antioxidants on sperm kinematic parameters after the freezing-thawing process and the post thawed fertility rate after the trans cervical artificial insemination. Methods: Ejaculates from Lacune rams were collected using artificial vagina, were pooled and diluted using based extender (Biladyl, Minitube) and based extender with antioxidants (Eugenol, Arginine). Trans-cervical artificial insemination was performed with frozen seminal material in straws. Result: No significant differences were observed in samples cryopreserved in a base extender and in the base extender supplemented with Arginine. In Eugenol supplemented extender the kinematic parameters were significantly higher for p=0.5 (30.15% progressive motility after thawing) than in the others groups (23.57%, 24.38%). The fertility rate was higher for the straws conserved in extender supplemented with Eugenol (80%) compared with the base extender (55.55%) and the base extender supplemented with Arginine (60%).