Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.

Extracellular vesicles (EVs) carry bioactive cargoes involved in the early preimplantation development. This study investigated the effects of EVs obtained from an oviductal epithelial cell (OEC) conditioned medium on the developmental competence of in parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) porcine embryos. The OEC‐EV‐treated group showed significant increases in blastocyst formation and hatching rates compared to the control group (40.8% ± 2.2% and 20.1% ± 2.1% vs. 24.9% ± 2.0% and 5.3% ± 1.1%; p < 0.05), respectively. The 7 day OEC‐EVs treatment group significantly increased blastocyst formation rate than the 3 day and 0 day‐groups (45.0 ± 0.8 vs. 33.0 ± 0.7 and 26.7 ± 0.5; p < 0.05), respectively. SCNT revealed that the OEC‐EV increased blastocyst formation rate compared to that of oviductal fluid EVs (OF‐EVs) (35.4% ± 1.4% vs. 29.3% ± 1.3%; p < 0.05). Reactive oxygen species levels, apoptosis, and blastocyst lipid content were significantly decreased in the OEC‐EVs group compared with the control group. OEC‐EV group showed a significantly decreased BAX and increased BCL2, SOD1, POU5F1, SOX2, NANOG, GATA6, PNPLA2, LIPE, and MGLL gene expression than the control group (p < 0.05). In conclusion, OEC‐EVs supplementation in embryo culture media improved the quality of porcine embryos, potentially helping porcine‐cloned embryonic development possibly through transfer of messenger RNA and proteins to the early embryos.

A cross-sectional study was conducted from March 2019 to February 2020 with the objective of identifying ixodid ticks and haemoparasites, in extensively managed livestock, in Alle district, Southwestern Ethiopia. The study area is assumed to be free from ticks, and there had been no diagnostic and treatment options for tick-borne diseases. Among 384 heads of cattle examined for tick infestation and haemoparasites, 139 (36.19%) were infested with one or more tick species and 25 (6.51%) were haemoparasitised. Two genera of ticks, Amblyomma and Rhipicephalus formerly (Boophilus), and four species (Amblyomma variegatum, Amblyomma lepidum, Rhipicephalus microplus, and Rhipicephalus annulatus) were identified. The haemoparasite identified was Babesia bovis. Among the risk factors, body condition score and season of the year were found to be significantly associated with tick infestation with x2 = 9.919, p > 0.05 and x2 = 6.216, p > 0.05 , respectively, at 95% CI. Tick infestation was found to be significantly associated with haemoparasitemia with x2 = 22.2 and p > 0.05 , at 95% CI. The finding of the current study is an alarm ring, as the veterinary service had been not considering any haemoparasitemia in the potential list of differential diagnosis and no treatment inputs have been availed for that purpose. Thus, it is recommended that the veterinary service delivery system in the area should take haemoparasites diagnosis and avail treatment alternatives, particularly tick-borne diseases. Furthermore, there should be a strategical approach in controlling tick-borne diseases in the area before the tick-borne diseases get prevalent and where the control after high prevalence could not be easy in extensive livestock management.

The objective of this study was to investigate the effect of milrinone supplementation as a phosphodiesterase 3A inhibitor during in vitro maturation (IVM) to coordinate the cytoplasmic and nuclear maturation of porcine oocytes and subsequent development of porcine cloned embryos. Brilliant cresyl blue (BCB)-stained (BCB +) oocytes, classified as well-developed, and BCB− oocytes were used in parthenogenesis (PA) and cloning, and their preimplantation development was compared. In PA embryos, BCB + oocytes had significantly higher rates of development than BCB− oocytes in terms of maturation (87.5 vs. 71.3%), cleavage (88.6 vs. 76.3%), and blastocyst development (34.3 vs. 25.3%) and also had higher cell numbers (46.9 vs. 38.9%), respectively (p &lt; 0.05). In cloned embryos, the BCB + group also had a significantly higher blastocyst formation rate than the BCB− group (30.6 vs. 20.1%; p &lt; 0.05). Supplementation with 75 μM milrinone during IVM of BCB− oocytes showed improvement in maturation and blastocyst development rates, which may be due to the coordinated maturation of the cytoplasm with the nucleus as an effect of milrinone. Moreover, the analysis of nuclear reprogramming via the examination of the expression levels of the reprogramming-related genes POU5F1, DPPA2, and NDP52IL in milrinone-supplemented BCB− oocytes showed higher expression levels than that in non-treated BCB− oocytes. These findings demonstrate that milrinone is useful in improving developmental competence in less competent oocytes during IVM and for proper nuclear reprogramming in the production of porcine cloned embryos by coordinating cytoplasmic and nucleus maturation.