Background: The poultry industry experiences genetic losses due to recurring infectious diseases, necessitating effective preservation strategies. Nitric oxide plays a crucial role in male reproduction, and optimal NO (nitric oxide) levels may enhance sperm viability. This study investigated the effects of SNAP (S-nitroso-Nacetylpenicillamine) on the longevity of rooster sperm. Methods: Semen was diluted with Beltsville Poultry Semen Extender-I containing 0 or 25 μM SNAP and stored at 10°C. Sperm motility and acrosome integrity were assessed at 1, 3, and 7 days. NO levels were quantified by DAF-FM diacetate and AI trials were evaluated by fertility and hatchability. Results: On day 1, sperm motility in the SNAP 25 μM-treated group was significantly higher than in the control. NO quantification confirmed that SNAP-treated semen exhibited higher NO levels. For fertilization and hatchability assessment, hens were divided into two groups based on the presumed duration sperm resided in sperm storage tubules. Before artificial insemination, the sperm was preserved at low temperature (10°C) to maintain viability. Fertilization rates were significantly higher in the SNAP-treated group in both short-term and long-term SST storage conditions. However, hatchability was only significantly improved in the SNAP-treated group when fertilization occurred after extended storage. Conclusions: These findings suggest that NO enhances sperm viability and fertility in poultry semen stored at low temperatures. SNAP 25 μM enhances AI efficiency by maintaining sperm viability and extending fertilization potential. Further research is needed to refine NO-based fertility enhancement strategies for avian species.
Background: Hanwoo cattle, an indigenous Korean breed, have become economically significant due to genetic improvements and large-scale farming. As individual cow value increases, understanding their unique physiology across different life stages is crucial for optimal health management. This retrospective study aimed to investigate serum biochemistry differences among non-pregnant, pregnant, and fattening female Hanwoo cattle and establish breed-specific reference intervals (RIs) for accurate health assessment, utilizing data obtained from routine veterinary care. Methods: Blood samples were collected from female Hanwoo cattle, categorized as pregnant (n = 12), non-pregnant (n = 25), and fattening (n = 11). Eighteen serum biochemical parameters were analyzed and descriptive statistics were calculated for each group. The new RIs in different reproductive status of female Hanwoo were established using the Reference Value Advisor program. Results: Significant differences based on reproductive status were identified in blood urea nitrogen (BUN), γ-glutamyl transferase (GGT), triglyceride (TG), glucose (GLU), and creatinine (CRE) levels. BUN, GGT, and TG levels were significantly higher in fattening cattle compared to pregnant and non-pregnant cows. GLU levels increased progressively across pregnant, non-pregnant, and fattening groups, while CRE levels were significantly higher in pregnant cows. Based on values of biochemical parameters, new RI were suggested for sixteen biochemical parameters, encompassing all three reproductive stages. Conclusions: This study established new RIs for female Hanwoo cattle across nonpregnant, pregnant, and fattening stages, providing a more accurate basis for health assessment and management. These findings will contribute to improved individual cow management, supporting genetic improvement efforts, and enhancing overall herd health in female Hanwoo cattle.
Background: Hybridization between closely related fish species can generate novel phenotypes that influence aquaculture performance. This study aimed to evaluate the morphological characteristics of hybrids between the two aquaculture-relevant flounder species Kareius bicoloratus and Platichthys stellatus using a hybrid index and a newly proposed resemblance p-value-based morphometric analysis, providing insights into hybrid resemblance patterns relative to their parental species. Methods: One-year-old individuals from the three genotype groups (K. bicoloratus, P. stellatus , and hybrid) were analyzed using a combination of traditional and trussbased morphometrics. From the full dataset, 77 morphological indices were extracted, including proportions, ratios, and angular measurements. The hybrid index was computed to quantify parental resemblance, while the delta resemblance value (ΔRV) was derived from Kruskal-Wallis test to assess statistical resemblance trends. One-way ANOVA and multiple comparison tests were used to determine statistical significance among groups. Results: Hybrid flounders exhibited a complex blend of parental and hybrid-specific traits, with morphological resemblance varying by trait category. Among the 77 morphological indices, 44 (57.1%) fell within the parental range, while 33 (42.9%) exceeded parental values, demonstrating transgressive segregation or heterosis in hybrid morphology. Morphometric resemblance patterns were trait-dependent: indices relative to total length or head length tended to resemble maternal species, whereas depth-related ratios and angular traits were more similar to father. Conclusions: The integration of H-index and ΔRV analysis provided a systematic and quantitative framework for assessing hybrid morphology, offering valuable insights into phenotypic expression of hybrids, with potential relevance to aquaculture.
Background: This study aimed to select high-quality spermatozoa by fresh domestic feline semen using sperm separation by magnetic activation (MASS), compared to density gradient centrifugation (DGC), and to evaluate cell quality after selection. Methods: Semen was collected from ten domestic felines by pharmacological sampling using dexmedetomidine and ketamine followed by urethral catheterization. The following parameters were analyzed: motility (computer assisted semen analysis), concentration (Neubauer chamber), semen morphology (humid chamber), and supravital test (eosin/nigrosine staining). In DGC, 20 × 106 spermatozoa were used in a gradient of Percoll at 90% and 45%, centrifuged at 900 g for 5 min, and the pellet was diluted in HEPES buffer. In MASS, 10 × 106 spermatozoa were diluted in HEPES buffer, centrifuged at 300 g for 10 min. The pellet was then resuspended in HEPES buffer with nanoparticles bound to annexin V, incubated for 15 min, and then filtered in the magnetic separation column. Results: The total abnormalities in the fresh semen were 47.9 ± 4.47%, with DGC and MASS being effective in reducing sperm abnormalities by 15.4 ± 0.95% and 29.80 ± 4.90%, respectively. Conclusions: This nanotechnological method is efficient in producing high-quality semen samples for assisted reproduction.
Background: Embryo implantation is a complex process regulated by interactions between endometrial epithelial and stromal cells. The endometrium plays a critical role in this process, providing a supportive environment for embryo attachment. However, conventional 2D cell culture models fail to fully replicate the complex 3D structure and cellular interactions of the endometrium. To overcome these limitations, 3D organoid models have been developed to better mimic the in vivo endometrial environment. Methods: In this study, a multicellular uterine organoid model was developed using porcine endometrial epithelial cells (pEECs) and porcine endometrial stromal cells (pESCs) to evaluate the effects of the endometrial environment on embryo implantation. First, single-cell endometrial organoids (pEOs) were formed by culturing pEECs in Matrigel, and their basic cellular characteristics were assessed. Then, a multicellular uterine organoid model was established by combining pEOs with pESCs. Finally, porcine embryos were co-cultured with this model to examine its effect on embryo attachment. Results: The multicellular uterine organoid model facilitated embryo attachment, demonstrating that the 3D structure and cellular interactions of the endometrium play a significant role in embryo implantation. The presence of both epithelial and stromal cells contributed to a more physiologically relevant environment that supported embryo adhesion. Conclusions: This study demonstrates that a multicellular uterine organoid model can serve as a useful in vitro system for porcine embryo implantation research. This model may contribute to a better understanding of embryo development and implantation mechanisms, with potential applications in regenerative medicine and biotechnology.
Background: The cumulus cells (CC) play an essential role in protecting oocytes and providing molecular signals for meiotic and cytoplasmic maturation. Gonadotropins stimulate CC proliferation, promote the release of factors that resume oocyte maturation, and activate small G proteins. Among these, Ras, a GTP-binding protein, participates in signaling pathways that regulate cell growth, division, and proliferation. This study aimed to investigate the changes in the Ras subfamily and gonadotropin receptor expression during porcine CC maturation. Methods: Cumulus-oocyte complexes (COCs) were incubated in a medium supplemented with follicular stimulating hormone (FSH), luteinizing hormone (LH), and epidermal growth factor (EGF) for 44 hours. CCs were collected from the COCs at 0, 22, and 44 hours, and mRNA expression levels of gonadotropin and growth factor receptors (FSHR, LHR, EGFR), Ras subfamily members (H-Ras, K-Ras, N-Ras, R-Ras), and Ras GTPases (RASA1, SOS1) were analyzed using quantitative RT-PCR. Results: The results revealed that LHR and R-Ras mRNA expression significantly increased only at 44 hours compared to the 0-hour group (p < 0.05). Conversely, RASA1 mRNA levels decreased significantly at the same time points. No significant changes were observed in H-Ras, K-Ras, N-Ras , or SOS1 expression. Conclusions: In conclusion, the observed increase in LHR and R-Ras and the decrease in RASA1 provide new insights into the molecular dynamics of Ras subfamily members during porcine CC maturation, contributing to a better understanding of the regulatory mechanisms underlying oocyte development.