Researchers have contributed by increasing our understanding of the factors affecting reproduction in beef, mainly physical health and nutrition aspects, which have been main concerns during decades. Animal welfare is of outmost relevance in all animal production systems and it is strongly associated to stress. Stress responses involve endocrine, paracrine and neural systems and the consequences of this stress on the reproductive efficiency of specifically, beef cattle and bulls, need to be highlighted. We, therefore, describe the fundamentals of stress and its quantification, focusing in beef herds, reviewing the highly valuable pieces of research, already implemented in this field. We examine major factors (stressors) contributing to stress in beef cattle and their effects on the animals, their reproductive performance and the success of reproductive biotechnologies. We include terms such as acclimatization, acclimation or temperament, very relevant in beef systems. We examine specifically the management stress due to handling, social environment and hierarchy or weaning effects; nutritional stress; and thermal stress (not only heat stress) and also review the influence of these stressors on reproductive performance and effectiveness of reproductive biotechnologies in beef herds. A final message on the attention that should be devoted to these factors is highlighted.

CASA kinetic parameters are often evaluated in a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed^®, BIOXcell^® or INRA96^®) on CASA kinetic parameters. Total and progressive motility, VCL, VAP, VCL, ALH, BCF, STR, LIN, and WOB were assessed. CASA kinetic parameters were preserved for up to 4–6 h post-ejaculation, except for AndroMed^®. Regardless of extender or temperature, motility decreased from 4–6 h up to 24 h, with the best values obtained with BIOXcell^® at 5 °C. Our results suggest that BIOXcell^® can preserve sperm motility for up to 6 h, either at 5 °C or room temperature, and also INRA96^® at room temperature, with motility assessments and the percentage of the most rapid sperms being the lowest with INRA96^® at 5 °C. The kinetic parameters decreased when analyses were performed at 24 h. Therefore, we suggest evaluating seminal quality as soon as possible, before 6 h after collection. These results help to fix adequate protocols for the short-term storage and shipment of bovine semen collected under field conditions.

Seminal parameters can be evaluated in situ, or samples can be delivered to a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed^®, BIOXcell^® or INRA96^®) on semen quality. Acrosome integrity, sperm viability and morphology, CASA-total and progressive motility, pH, and colony-forming units were assessed. Semen quality was preserved for up to 4–6 h post-ejaculation, except for INRA96^® at 5 °C. Regardless of extender or temperature, motility decreased from 4 to 6 h up to 24 h, with the best values obtained with BIOXcell^® at 5 °C. pH differed from 4 to 6 h up to 24 h, acidifying when stored at room temperature. Microbiological load was stable over time with AndroMed^® and BIOXcell^®, and increased at room temperature with INRA96^®. Our results suggest that AndroMed^® and BIOXcell^® can preserve semen quality for up to 6 h, either at 5 °C or room temperature, while INRA96^® only at room temperature. These results help to fix adequate protocols for short-term storage and shipment of bovine semen collected under field conditions.

Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016–2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production (p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling (p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; (p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea (p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients’ metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy.

Barbary sheep (Ammotraguslervia) are considered vulnerable species by the World Conservation Union (IUCN). The purpose of this study was to describe the basic characteristics of fresh semen, test the efficacy of commercial extender Triladyl, and collect necessary data that may help to create a frozen semen bank for the Barbary sheep in Spain. A total of 21 ejaculates were collected by rectal-probe electroejaculation from one dominant (D) and three minor (M) adult males housed in the Madrid Zoo. After ejaculation, semen volume, concentration, and mass motility were assessed. Remaining raw semen was diluted at 37°C with TRIS-based extender Triladyl (Minitüb, Tiefenbach, Germany) and 20% egg yolk to a final concentration of 200 × 106 sperm per mL. Diluted samples were kept at 5°C for 4 h and then loaded into 0.25-mL French straws, frozen at 5 cm above liquid nitrogen (LN2) for 10 min and then plunged into LN2. Samples were thawed in a water bath at 37°C for 30 s. Post-thaw semen survival was evaluated by sperm motility (%M), quality of movement (Q), normal acrosome status (%NAS), normal sperm morphology (%NOR), membrane integrity (hypo-osmotic test; %HOST), and sperm viability (eosin-nigrosin vital staining; %V), and were compared with the same parameters in the fresh semen. Data between D and M males were analyzed by one way ANOVA. Mean volume of ejaculates, total sperm concentration and mass motility of raw semen were respectively; 5.2 ± 1.56 mL, 2800.0 ± 1290.5 × 106 and 3.4 ± 0.4 for the D male, and 3.5 ± 3.2 mL, 251.2 ± 103.9 × 106, and 1.88 ± 1.4 for M males (P < 0.05). Remaining semen parameters evaluated in raw semen showed no differences between D and M males. However, post-thaw semen quality was significantly (P < 0.05) reduced in all analyzed parameters except %NAS and %NOR in M males groups as compared to the D male (Table 1). It can be concluded that Barbary sheep raw semen collected by electroejaculation is of sufficient quality to be used in an artificial insemination program and can be successfully frozen in commercially available Triladyl extender. However, the post-thaw viability of semen may considerably depend on the male reproductive status in the flock. Table 1. Characteristics of fresh and cryopreserved Barbary sheep semen This work was supported by CAM 07B/007/1999 (Analysis of ejaculate traits and development of methods of semen preservation in wild ungulates from the Madrid Zoo).