Mrenoshki Slavcho, Temerario Letizia, Mastrorocco Antonella, Visci Grazia, Notario Elisabetta, Marzano Marinella, Martino Nicola Antonio, Mrenoshki Daniela, Lacalandra Giovanni Michele, Pesole Graziano, Dell'Aquila Maria Elena
Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, North Macedonia.
Department of Biosciences, Biotechnology and Environment, University of Bari Aldo Moro, 70125 Bari, Italy.
Animals (Basel). 2025 Jul 2;15(13):1951. doi: 10.3390/ani15131951.
The aims of the present study were to analyze the taxonomic profile and to evaluate the functional effects of sheep FF cultivable microbiota on prepubertal lamb oocytes PLOs developmental potential. Ovarian FFs were recovered from slaughtered adult sheep via the aspiration of developing follicles and used for microbiota propagation. Bacterial pellets underwent 16S rRNA gene sequencing and targeted culturomics, whereas cell-free supernatants were used as supplements for the in vitro maturation (IVM) of slaughtered PLOs. For the first time, bacteria presence in adult sheep FF was detected, with the first report of subsp. (as a species) and (as a genus and species) in either animal or human FF. The short- and long-term effects of bacterial metabolites on PLO maturation and embryonic development were demonstrated. As short-term effects, the addition of FF microbiota metabolites did not affect the oocyte nuclear maturation and mitochondria distribution pattern, except in one of the examined supernatants, which reduced all quantitative bioenergetic/oxidative parameters. As long-term effects, one of them reduced the total cleavage rate after in vitro embryo culture (IVC). In conclusion, microbiota/bacteria are present in adult sheep FF and may influence reproductive outcomes in vitro. Future studies may reveal the beneficial in vitro effects using the microbiome from preovulatory follicles.
本研究的目的是分析绵羊卵泡液（FF）可培养微生物群的分类学特征，并评估其对青春期前羔羊卵母细胞（PLOs）发育潜能的功能影响。通过抽吸发育中的卵泡，从屠宰的成年绵羊中获取卵巢FF，并用于微生物群的增殖培养。对细菌沉淀进行16S rRNA基因测序和靶向培养组学分析，而无细胞上清液则用作屠宰后PLOs体外成熟（IVM）的补充剂。首次在成年绵羊FF中检测到细菌的存在，首次报道了亚种（作为一个物种）和（作为一个属和物种）存在于动物或人类的FF中。证明了细菌代谢产物对PLO成熟和胚胎发育的短期和长期影响。作为短期影响，添加FF微生物群代谢产物不影响卵母细胞核成熟和线粒体分布模式，但在所检测的一种上清液中除外，该上清液降低了所有定量生物能量/氧化参数。作为长期影响，其中一种代谢产物降低了体外胚胎培养（IVC）后的总卵裂率。总之，成年绵羊FF中存在微生物群/细菌，可能会影响体外生殖结果。未来的研究可能会揭示使用排卵前卵泡的微生物组在体外产生的有益效果。
