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We evaluated the quality and fertilizing ability of frozen–thawed porcine sperm that were selected using a commercially available device (MIGLIS, Menicon Life Science) consisting of three parts: an outer lid, an inner lid, and a tube. Firstly, to determine an adequate concentration of caffeine for separation, frozen–thawed sperm were incubated with different concentrations of caffeine (0, 1, 2.5, 5, and 10 mM) in a MIGLIS device. To determine the appropriate incubation time for separating sperm in the MIGLIS device, frozen–thawed sperm were incubated with 2.5 mM caffeine for 5, 10, 15, or 20 min. To evaluate the fertilization and embryo development of oocytes fertilized with frozen–thawed sperm separated into two regions (outer and inner) in the MIGLIS device, the separated sperm from the three boars was used to fertilize in vitro‐matured oocytes and cultured in vitro for 7 days. Sperm quality parameters of sperm collected from the inner tube after incubation with 2.5 mM caffeine were superior to sperm incubated without caffeine. Moreover, sperm collected from the inner tube after incubation for 10 min had a higher progressive motility. The rate of blastocyst produced from spermatozoa collected from the inner tube after incubation with 2.5 mM caffeine for 10 min significantly increased compared to that produced from spermatozoa from the outer tube, regardless of the boar. In conclusion, sperm sorting using the MIGLIS device may be useful for separating high‐quality sperm after incubation with 2.5 mM caffeine for 10 min to improve blastocyst formation. [ABSTRACT FROM AUTHOR]

Sterilization of the culture medium using ultraviolet (UV)‐C reduces the potential adverse effects of microorganisms and allows for long‐term use. In the present study, we investigated the effects of a medium directly irradiated with UV‐C prior to in vitro culture on the development and quality of porcine in vitro‐fertilized embryos and the free amino acid composition of the culture media. The culture media (porcine zygote medium [PZM‐5] and porcine blastocyst medium [PBM]) were irradiated with UV‐C at 228 and 260 nm for 1 and 3 days, respectively. Next, the culture media were irradiated with UV‐C at 228 nm for 3, 7, or 14 days. After in vitro fertilization, the embryos were cultured in the UV‐C‐irradiated media for 7 days. Free amino acid levels in culture media irradiated with 228 and 260 nm UV‐C for 3 days were analysed. The blastocyst formation rate of embryos cultured in media irradiated with 260 nm UV‐C for 3 days was significantly lower than that of embryos cultured in non‐irradiated control media. However, 228 nm UV‐C irradiation for up to 14 days did not affect blastocyst formation rates and quality in the resulting blastocysts. Moreover, 260 nm UV‐C irradiation significantly increased the taurine concentration in both culture media and decreased methionine concentration in the PBM. In conclusion, UV‐C irradiation at 228 nm before in vitro culture had no detrimental effects on embryonic development. However, 260 nm UV‐C irradiation decreased embryo development and altered the composition of free amino acids in the medium. [ABSTRACT FROM AUTHOR]

This study examined the effects of ergothioneine (EGT) supplementation as an antioxidant on the quality of boar spermatozoa when using liquid and frozen preservation methods. In the first experiment, boar semen was preserved in an extender supplemented with 0, 50, 100 and 200 µM EGT, at 15 °C, part of the samples for one and another part for three weeks. In comparison with the control (without EGT), EGT supplementation at 100 µM significantly increased the percentage of total motility of spermatozoa that were preserved as a liquid both for one and three weeks (P < 0.05). EGT supplementation did not affect the quality of preserved spermatozoa, irrespective of the EGT concentration. In the second experiment, semen was frozen and thawed in the freezing extender supplemented with 0, 50, 100 and 200 µM EGT. In comparison with the control, the 100 µM EGT supplementation significantly increased the percentages of total and progressive motility of frozen-thawed spermatozoa (P < 0.05). EGT (100 µM) supplementation did not affect the viability, the plasma membrane integrity, or the acrosomal integrity of frozen-thawed spermatozoa. These findings indicate that supplementing extenders with 100 µM EGT may improve the motility of boar sperm in both liquid and freezing preservation methods. [ABSTRACT FROM AUTHOR]