Spermatozoa are highly specialized male cells that are characterized by a unique ability to move, which is a critical factor for successful fertilization. The relative simplicity of motility assessment, especially in livestock, has made it a widely used parameter for evaluating ejaculate quality or cryopreserved semen in the clinical field, and an advanced tool in reproductive physiology and toxicology research. Technological advances in image analysis and computational methods have substantially increased its accuracy through the use of computer-assisted sperm analysis (CASA) to minimize subjective bias in motility assessments. Nevertheless, this more objective method still presents some significant challenges, including variability in the sample preparation, imaging conditions, and analytical parameters. These issues contribute to inconsistency and impair the reproducibility and comparability of data between laboratories. The implementation of standardized protocols, combined with comprehensive training and rigorous evaluation, can serve to mitigate some of the emerging inconsistencies. In addition, the in vitro conditions under which CASA analyses are performed often differ significantly from the natural environment of the female reproductive tract in vivo. This review discusses the methodologies, critical issues, and limitations of sperm motility analyses using CASA, with a particular focus on the boar as an important agricultural and biomedical model species in which this system is widely used.

The study was conducted during the summer season (June–August 2020). Two hundred sixty-four 5-week-old sexed Muscovy ducklings were randomly divided into four equal experimental groups by housing system and by gender. Each group had three replicates (22 birds/replicate) in a randomized design experiment. Regarding the hematological traits, the volume of leukocytes was higher in the D group (by 0.34 × 10⁹/L; p < 0.05) than in the S group. Furthermore, body temperature was found to be higher in ducks (by 0.84 °C; p < 0.05) and in the D group (by 0.5 °C; p < 0.05) in comparison with drakes and birds from the S group. Considering relative brain weight, drakes had higher values than ducks (by 0.56 g; p < 0.05), and birds from the S group also manifested higher values (by 0.78 g; p < 0.05). In terms of bone quality, there were no differences in studied parameters of tibia and femur bones regarding housing systems. The results provide valuable evidence of differences in the fattening of intensively bred Muscovy ducks within the housing system but also regarding gender.

The present study aimed to demonstrate the dependence of meiotic maturation in pig oocytes on the activity of the protease complex proteasome. The proteasome inhibitor MG132 blocked the exit of maturing pig oocytes from metaphase I stage. Seventy-five per cent of the oocytes were blocked at metaphase I when they were cultured with 10 μM MG132. The blocking effect of MG132 was expressed only when the oocytes were exposed to an inhibitor before the 18th hour of in vitro culture. The effects of MG132 are fully reversible. However, a significant proportion of oocytes (46%) cultured for 48 h in MG132-supplemented medium and then for 24 h in MG132-free medium did not block meiosis at the stage of metaphase II and underwent spontaneous parthenogenetic activation. On the basis of our data we can conclude that exit from the metaphase I stage of meiosis is proteasome-dependent in pig oocytes matured in vitro. On the other hand, our data also indicate that other proteasome-independent events are involved in regulating the exit from metaphase I.

Antiepileptic drugs (e.g., carbamazepine; CBZ) are widely prescribed for various conditions beyond epilepsy, including neurologic and psychiatric disorders. These medications can have both favorable and unfavorable impacts on mood, anxiety, depression, and psychosis. CBZ has been found at low concentrations (in the unit of nanograms per liter) in rivers, surface water, and even drinking water. As a result, when reclaimed wastewater is used for irrigation in agricultural ecosystems, CBZ can be reintroduced into the environment. That is why we tested different doses of CBZ in rabbits’ feed as the meat is consumed in every community, has no religious barriers, and the potential risk of consuming meat which has been exposed to CBZ treatment is not known. Also, the evidence of the effect of CBZ on rabbits is missing. Mainly, the CBZ doses affected the count of leukocytes and other blood traits, meaning the higher the dose, the higher the reduction. Moreover, there were only low amounts of CBZ in rabbits’ meat or tissues when they were exposed to the treatment.

Calcineurin is required for oocyte exit from meiotic block in metaphase II (MII) stage in invertebrates and also in lower vertebrates. However, the role of calcineurin in mammalian oocyte activation is still unclear. The aim of this study was to determine whether calcineurin is involved in the processes regulating porcine oocyte activation. Indirect immunofluorescence demonstrated localization of both calcineurin subunits, CnA and CnB, especially in the cortex area of MII oocytes, in vitro fertilized and also parthenogenetically activated oocytes. After activation, the fluorescence intensity of the protein in the cortex area of oocytes remains unchanged; the protein calcineurin in the cytoplasm was recorded mainly around the pronuclei. Treatment of matured oocytes with calcineurin inhibitors, cyclosporin A (CsA) and hymenistatin I (HS-I), followed by activation with calcium ionophore A23187, significantly decreased the rate of activated oocytes compared to oocytes that were treated only with calcium ionophore (Ca-Io), (CsA+Ca-Io 25.0% v. Ca-Io 83.3%; HS-I+Ca-Io 32.5% v. Ca-Io 85.0%). Compared to the control, CsA treatment of matured oocytes followed by activation with Ca-Io did not affect the activity level of metaphase-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) in activated oocytes evaluated by kinase activity assay. Simultaneous staining of calcineurin and cortical granule content in matured oocytes showed that calcineurin distributed in the cortical area of the oocyte has not been colocalized with cortical granules content. On the other hand, the calcineurin inhibition before parthenogenetic activation leads to a reduction of the cortical reaction level compared to oocytes that were not treated with CsA (complete exocytosis: CsA+Ca-Io 2.6% v. Ca-Io 83.9%; sum of cortical granule brightness: CsA + Ca-Io 0.69 v. Ca-Io 0.15). Our results showed that calcineurin is involved in the process of pig oocyte activation and cortical granule exocytosis; however this regulation seems to be MPF and MAPK independent.