Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia.
Department of Fruit Growing, Viticulture and Enology, Faculty of Horticulture and Landscape Engineering, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia.
Molecules. 2019 Nov 27;24(23):4329. doi: 10.3390/molecules24234329.
Male subfertility is a global issue in human reproduction as well as in animal reproduction. Bacterial infection and semen contamination are still widely overlooked. As the collection of ejaculates is not a sterile process, it is necessary to add antimicrobial agents to avoid a possible depreciation of semen samples. As traditionally used antibiotics have been questioned because of an ever-increasing bacterial resistance, natural bioactive molecules could offer an alternative because of their antibacterial and antioxidant properties. As such, we decided to compare the effects of selected natural biomolecules (resveratrol-RES, quercetin-QUE and curcumin-CUR) with routinely used antibiotics in animal biotechnologies (penicillin-PEN, gentamicin-GEN and kanamycin-KAN) on the rabbit sperm vitality in the presence of . Changes in the sperm structural integrity and functional activity were monitored at 0, 2, 4 and 6 h. Computer-assisted sperm analysis (CASA) was used for the assessment of spermatozoa motility. Production of reactive oxygen species (ROS) was evaluated using chemiluminiscence, while the mitochondrial membrane potential (ΔΨm) was examined using the JC-1 dye. Finally, the sperm chromatin dispersion (SCD) test was used to assess DNA fragmentation, and changes to the membrane integrity were evaluated with the help of annexin V/propidium iodide. The motility assessment revealed a significant sperm motility preservation following treatment with GEN ( < 0.001), followed by PEN and CUR ( < 0.01). QUE was the most capable substance to scavenge excessive ROS ( < 0.001) and to maintain ΔΨm ( < 0.01). The SCD assay revealed that the presence of bacteria and antibiotics significantly ( < 0.05) increased the DNA fragmentation. On the other hand, all bioactive compounds readily preserved the DNA integrity ( < 0.05). In contrast to the antibiotics, the natural biomolecules significantly maintained the sperm membrane integrity ( < 0.05). The microbiological analysis showed that GEN ( < 0.001), KAN ( < 0.001), PEN ( < 0.01) and CUR ( < 0.01) exhibited the strongest antibacterial activity against . In conclusion, all selected biomolecules provided protection to rabbit spermatozoa against deleterious changes to their structure and function as a result of contamination. Therefore, administration of RES, QUE and/or CUR to rabbit semen extenders in combination with a carefully selected antibacterial substance may be desirable.
男性不育症是人类生殖和动物生殖中的一个全球性问题。细菌感染和精液污染仍然被广泛忽视。由于精液采集不是无菌过程，因此有必要添加抗菌剂以避免精液样本可能的贬值。由于传统使用的抗生素由于细菌耐药性的不断增加而受到质疑，因此具有抗菌和抗氧化特性的天然生物活性分子可能是一种替代方法。因此，我们决定比较所选天然生物分子（白藜芦醇-RES、槲皮素-QUE 和姜黄素-CUR）与动物生物技术中常用的常规抗生素（青霉素-PEN、庆大霉素-GEN 和卡那霉素-KAN）对兔精子活力的影响。在存在的情况下。在 0、2、4 和 6 h 监测精子结构完整性和功能活性的变化。使用计算机辅助精子分析 (CASA) 评估精子运动。使用化学发光法评估活性氧 (ROS) 的产生，同时使用 JC-1 染料检查线粒体膜电位 (ΔΨm)。最后，使用精子染色质分散 (SCD) 试验评估 DNA 碎片化，并用 Annexin V/碘化丙啶评估膜完整性的变化。运动评估显示，用 GEN 处理后精子运动显著保存（<0.001），其次是 PEN 和 CUR（<0.01）。QUE 是清除过多 ROS 的最有效物质（<0.001），并保持 ΔΨm（<0.01）。SCD 试验表明，细菌和抗生素的存在显著（<0.05）增加了 DNA 碎片化。另一方面，所有生物活性化合物都容易地保持了 DNA 完整性（<0.05）。与抗生素不同，天然生物分子显著地保持了精子膜的完整性（<0.05）。微生物分析表明，GEN（<0.001）、KAN（<0.001）、PEN（<0.01）和 CUR（<0.01）对具有最强的抗菌活性。总之，所有选定的生物分子都为兔精子提供了保护，防止其结构和功能因污染而发生有害变化。因此，在兔精液稀释剂中添加 RES、QUE 和/或 CUR 并结合精心选择的抗菌物质可能是可取的。
