PGCs offer a viable means for cryobanking both female and male avian genetic material. These early-migrating cells ultimately give rise to eggs and sperm cells, making them valuable for long-term conservation of poultry genetic resources. In this study, PGCs from the endangered Oravka chicken, a local breed native to Slovakia, were isolated, propagated in vitro, characterized, and cryopreserved. A total of 18 stable PGC lines were successfully established, comprising 12 female and 6 male lines. Characterization using immunohistochemistry, gene expression analysis, karyotyping, cell proliferation assays, TEM, and in vivo migration assays confirmed the identity and developmental competency of the in vitro cultured PGCs. A total of 108 cryopreserved samples were deposited into the gene bank. This study reports the first successful derivation, in vitro culture and cryobanking of stable PGC lines from the Oravka breed, providing a foundational resource for the conservation and future restoration of this distinctive local genetic lineage.

Extreme weather impacts the ecological niches of fungi, altering mycotoxin risks in wildlife. We analyzed mycotoxin carry-over into European fallow deer (Dama dama) milk across seasons and assessed how drought influences the shift from Fusarium to Aspergillus mycotoxins and affects physiological resilience. Samples were collected during 2021/2022 and a drought-stricken 2022/2023 from South Transdanubia and Northeastern Hungary. Aflatoxin B1/M1 (AFB1/AFM1), Fumonisin B1 (FB1), Deoxynivalenol (DON), Zearalenone (ZEN), and Body Condition Scores (BCS) were measured to evaluate the impact of exposure on health status. The severe drought significantly altered the mycotoxin profile: ZEN levels declined significantly (from a median of 0.28 to 0.00 ng/mL), consistent with the moisture requirements of Fusarium graminearum, whereas DON concentrations increased. Concurrently, AFM1 persisted, exhibiting increased variance and extreme outliers in the maize-dominated South Transdanubian region. Distinct pharmacokinetic patterns were observed, and positive correlations were observed between milk and feces for lipophilic toxins, validating milk as a possible biomarker. Hydrophilic DON showed no correlation despite its accumulation. Emergence of “Poor” BCS group carrying loads supports “condition-dependent foraging” hypothesis, as stressed individuals are forced to consume contaminated resources, exacerbating oxidative stress and metabolic deficits.

MicroRNAs (miRNAs) are short, non-coding RNA molecules that play a crucial role in regulating various biological processes by influencing post-transcriptional gene expression and gene silencing. Background/Objectives: In this study, rabbit embryos were utilised as a model system to investigate potential biomarkers relevant to human embryo development. Seven microRNAs (miRNAs) identified in the embryo culture medium were evaluated as biomarkers by analysing the correlation between their expression levels and the developmental quality of rabbit embryos at days 4 and 6. Methods: We analysed the expression of seven development-specific miRNAs (miR-24-3p, miR-28-3p, miR-103a-3p, miR-181a-5p, miR-191-5p, miR-320a-3p, miR-378a-3p) in 4-day-old and 6-day-old rabbit embryos, along with their culture media. Results: Our findings revealed significant differences in the expression levels of these miRNAs between the 4-day-old and 6-day-old embryos. On the other hand, the expression patterns observed in the culture medium samples showed less variation between the two age groups. Nonetheless, analysis of miRNA expression profiles in the spent culture medium from individually cultured embryos enabled the identification of lower-quality embryos, characterised by smaller size and impaired or delayed development. Conclusions: The detection of these miRNAs in embryo culture medium may serve as a reliable indicator of successful progression to the blastocyst stage. Our experimental results identified specific miRNAs whose expression profiles differ according to embryonic stage and quality, thereby reflecting key developmental milestones. Notably, the detectability of these miRNAs in the medium—without prior RNA isolation—indicates their active secretion into the extracellular environment. By synthesising our findings with the existing literature, we refined a panel of miRNAs essential for the development of implantation-competent embryos in both rabbits and humans. Consequently, we developed a non-invasive assay for predicting implantation and pregnancy outcomes, which may have significant applications in human reproductive medicine.