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Chilled pork is frequently contaminated with Pseudomonas fragi and Pseudomonas fluorescens. In this study, the bactericidal efficacy and mechanisms of non-electrolytic slightly acidic hypochlorous water (NE-SAHW) against two strains of these two species were evaluated. The results showed that the antibacterial efficacy of NE-SAHW was positively correlated with the concentration level of NE-SAHW and negatively correlated with the initial populations of the strains. The strains of small populations were completely inhibited when provided with each level of NE-SAHW. The killed cells of P. fragi were 0.94, 1.39, 4.02, and 5.60 log10 CFU/mL, respectively, and of P. fluorescens they were 1.21, 1.52, 4.14, and 5.74 log10 CFU/mL, respectively, when the initial populations of the strains were at high levels (about 7 log10 CFU/mL). Both strains were completely killed within 12 s with the available chlorine concentration (ACC) of 50 mg/L of NE-SAHW. Morphological changes in both cells were observed by using a Scanning Electron Microscope (SEM) and it was discovered that the cell membranes were damaged, which led to the leakage of the intracellular substances, including K+, nucleic acid, and protein. In terms of the Fourier Transform Infrared Spectroscopy (FTIR) results, NE-SAHW destroyed the structures of membrane proteins and cell structure proteins, and influenced the composition of polysaccharides. The bacteria were definitely dead after treatment by NE-SAHW compared to the control according to the results of flow cytometry. These results demonstrated the potential bactericidal property of NE-SAHW when applied to the meat and other food sterilization industries.

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]

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]

The oocyte maturation process requires a high supply of energy, which generates reactive oxygen species (ROS), adversely affecting oocyte and embryo development. Balancing ROS by antioxidant supplementation is essential for maintaining oocyte maturation and embryonic quality in vitro. This study aimed to evaluate the impact of four antioxidants: β‐mercaptoethanol (β‐ME), chlorogenic acid (CGA), curcumin and sericin, when applied individually or in combinations, during oocyte maturation on development of porcine oocytes. Cumulus–oocyte complexes were collected, cultured in maturation medium supplemented with antioxidants for 44 hr and subsequently subjected to in vitro fertilization (IVF) and culture for 7 days. Combining all four (β‐ME + CGA + curcumin + sericin) or three (β‐ME + CGA + curcumin) antioxidants increased blastocyst formation rates. However, sericin supplementation alone, or in combination with β‐ME or CGA, failed to improve blastocyst formation rates. The total cell numbers of blastocysts from the group supplemented with three antioxidants (β‐ME + CGA + curcumin) were significantly higher than those from the other groups, except for the curcumin‐supplement group. There were no differences in the maturation rates and proportions of oocytes with fragmented DNA between the antioxidant‐supplemented and the non‐supplemented control groups. In conclusion, supplementation with three antioxidants (β‐ME + CGA + curcumin) during the maturation culture enhanced blastocyst formation and improved blastocyst quality. [ABSTRACT FROM AUTHOR]

Background: Increasing the permeability of the zona pellucida (ZP) of oocytes before CRISPR/Cas9 electroporation may improve the efficiency of gene editing; however, the effects of this approach on subsequent developmental processes are unclear. In this study, the effects of ZP treatment before electroporation on embryonic development and gene editing in porcine embryos were evaluated. Methods: The ZP of zygotes was weakened or removed by exposure to 0.5% actinase E, followed by electroporation of the Cas9 protein with guide RNA targeting GGTA1. Results: The blastocyst formation rate of ZP‐free zygotes after electroporation was significantly lower (p < 0.05) than that of ZP‐intact zygotes. The mutation rate in blastocysts from ZP‐weakened zygotes was similar to that in ZP‐intact zygotes, whereas ZP removal increased the mutation rate. The mutation efficiency in blastocysts from electroporated zygotes did not differ among ZP treatment groups. Conclusions: Our results indicate that weakening the ZP does not affect the developmental competence, mutation rate, or mutation efficiency of electroporated zygotes, whereas ZP removal has a detrimental effect on embryonic development but may increase the mutation rate. [ABSTRACT FROM AUTHOR]