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Dry-cured pork neck samples were stored at 10°C for 90 days under vacuum packaging (VP) or modified atmosphere packaging (MAP; 25% CO2+75% N2) conditions. The pH, moisture, water activity, total aerobic bacteria, and Enterobacteriaceae counts of dry-cured pork neck products with MAP were significantly lower than those with VP (p60 days of storage were lower than those at Day 1. In conclusion, despite presenting higher lipid oxidation, the samples stored in packages containing 25% CO2 for 90 days at 10°C have lower bacterial counts than vacuum-packed samples. Therefore, further studies should be performed on the packaging of dry-cured meat at adjusted concentrations of CO2.

The aim of this study was to investigate the total phenolic content, total flavonoid contents, antioxidant activity and antimicrobial activity of ethanolic extract from stems (S) and leaves (L) of Impatiens balsamina L. (Balsaminaceae), which were harvested in Korea on March 10, 2011 (S1 and L1), May 14, 2011 (S2 and L2), and July 5, 2011 (S3 and L3), respectively. Our results revealed that the total phenolic (79.55–103.94 mg CE/g extract) and flavonoid (57.43–104.28 mg QE/g extract) contents of leaf extract were higher (p < 0.01) than those of stem extract. Leaf extracts (L1, L2, and L3) exhibited stronger (p < 0.01) free radical scavenging activity (66.06, 63.71, and 72.19%, respectively) than that of the positive control. In terms of antimicrobial activity, leaf extracts showed higher inhibitory effects against microorganisms than those of stem extracts (S1, S2, and S3). Among the leaf extracts at different harvest times, L3 showed the greatest antimicrobial activity against both Gram negative and Gram positive strains. From these results, the leaf extract from I. balsamina L. might be a valuable bioactive resource, and would seem to be applicable as a natural antioxidant in food preservation.

Increasing litter size is of great interest to the pig industry. DNA methylation is an important epigenetic modification that regulates gene expression, resulting in livestock phenotypes such as disease resistance, milk production, and reproduction. We classified Berkshire pigs into two groups according to litter size and estimated breeding value: smaller (SLG) and larger (LLG) litter size groups. Genome-wide DNA methylation and gene expression were analyzed using placenta genomic DNA and RNA to identify differentially methylated regions (DMRs) and differentially expressed genes (DEGs) associated with litter size. The methylation levels of CpG dinucleotides in different genomic regions were noticeably different between the groups, while global methylation pattern was similar, and excluding intergenic regions they were found the most frequently in gene body regions. Next, we analyzed RNA-Seq data to identify DEGs between the SLG and LLG groups. A total of 1591 DEGs were identified: 567 were downregulated and 1024 were upregulated in LLG compared to SLG. To identify genes that simultaneously exhibited changes in DNA methylation and mRNA expression, we integrated and analyzed the data from bisulfite-Seq and RNA-Seq. Nine DEGs positioned in DMRs were found. The expression of only three of these genes (PRKG2, CLCA4, and PCK1) was verified by RT-qPCR. Furthermore, we observed the same methylation patterns in blood samples as in the placental tissues by PCR-based methylation analysis. Together, these results provide useful data regarding potential epigenetic markers for selecting hyperprolific sows.

Improvement in litter size has become of great interest in the pig industry because fecundity is directly related to sow reproductive life. Improved reproduction has thus been achieved by elucidating the molecular functions of genes associated with fecundity. In the present study, we identified differentially expressed genes (DEGs) via transcriptomic analysis using RNA-sequencing (RNA-Seq) in Berkshire pig placentas from larger (LLG, mean litter size >12) and smaller (SLG, mean litter size < 6.5) litter size groups. In total 588 DEGs were identified (p < 0.05, > 1.5-fold change), of which 98 were upregulated, while 490 were downregulated in the LLG compared with the SLG. Gene Ontology (GO) enrichment was also performed. We concluded that 129 of the 588 DEGs were closely related to litter size according to reproduction related genes selected based on previous reports, as 110 genes were downregulated and 19 upregulated in the LLG compared with the SLG. RT-qPCR utilizing specific primers targeting the early growth response 2 (EGR2), pheromaxein c subunit (PHEROC) and endothelial lipase (LIPG) genes showed high accordance with RNA-Seq results. Furthermore, we investigated the upstream regulators of these three genes in the placenta. We found that WNT9B, a Wnt signaling pathway molecule, and IL-6, known inducers of EGR2 and LIPG, respectively, were significantly increased in LLG compared with SLG. We believe that the induction of IL-6 and LIPG may play an important role in increasing nutrition supply through the placenta from the sow to the piglet during gestation. These results provide novel molecular insights into pig reproduction.

A previous study highlighted that mastoparan V1 (MP-V1), a mastoparan from the venom of the social wasp Vespula vulgaris, is a potent antimicrobial peptide against Salmonella infection, which causes enteric diseases. However, there exist some limits for its practical application due to the loss of its activity in an increased bacterial density and the difficulty of its efficient production. In this study, we first modulated successfully the antimicrobial activity of synthetic MP-V1 against an increased Salmonella population using protease inhibitors, and developed an Escherichia coli secretion system efficiently producing active MP-V1. The protease inhibitors used, except pepstatin A, significantly increased the antimicrobial activity of the synthetic MP-V1 at minimum inhibitory concentrations (determined against 106 cfu/mL of population) against an increased population (108 cfu/mL) of three different Salmonella serotypes, Gallinarum, Typhimurium and Enteritidis. Meanwhile, the E. coli strain harboring OmpA SS::MP-V1 was identified to successfully secrete active MP-V1 into cell-free supernatant, whose antimicrobial activity disappeared in the increased population (108 cfu/mL) of Salmonella Typhimurium recovered by adding a protease inhibitor cocktail. Therefore, it has been concluded that our challenge using the E. coli secretion system with the protease inhibitors is an attractive strategy for practical application of peptide toxins, such as MP-V1.

This study was performed to investigate the combined effect of dietary supplementation of gallic and linoleic acid (GL) on the antioxidative effect and quality of thigh meat from broilers. Broilers received 3 dietary treatments: i) commercial finisher diet (control), ii) 0.5% GL (gallic:linoleic acid = 1 M:1 M), and iii) 1.0% GL during the 22 to 36 d. The pH value of broiler thigh meat was increased by GL supplementation. Water holding capacity of the thigh meat was enhanced by the 1.0% dietary GL supplementation. Antioxidative effect (total phenolic content, DPPH radical scavenging activity, ABTS+ reducing activity, reducing power, and TBARS value) in the thigh from the broilers improved significantly with 1.0% GL. Linoleic acid and docosahexaenoic acids were higher in the broilers fed both levels of dietary GL. However, volatile basic nitrogen content and microbiological quality was not shown to be different between control and treated group. Results indicate that 1.0% dietary supplementation of GL can improve the antioxidant activity of broiler thigh meat and may enhance the meat quality.

Changes in meat quality traits are strongly associated with alterations in postmortem metabolism which depend on genetic variations, especially nonsynonymous single nucleotide variations (nsSNVs) having critical effects on protein structure and function. To selectively identify metabolism-related nsSNVs, next-generation transcriptome sequencing (RNA-Seq) was carried out using RNAs from porcine liver, which contains a diverse range of metabolic enzymes. The multiplex SNV genotyping analysis showed that various metabolism-related genes had different nsSNV alleles. Moreover, many nsSNVs were significantly associated with multiple meat quality traits. Particularly, ch7:g.22112616A>G SNV was identified to create a single amino acid change (Thr/Ala) at the 145th residue of H1.3-like protein, very close to the putative 147th threonine phosphorylation site, suggesting that the nsSNV may affect multiple meat quality traits by affecting the epigenetic regulation of postmortem metabolism-related gene expression. Besides, one nonsynonymous variation, probably generated by gene duplication, led to a stop signal in porcine testicular carbonyl reductase (PTCR), resulting in a C-terminal (E281-A288) deletion. Molecular docking and energy minimization calculations indicated that the binding affinity of wild-type PTCR to 5α-DHT, a C(21)-steroid, was superior to that of C-terminal-deleted PTCR or human carbonyl reductase, which was very consistent with experimental data, reported previously. Furthermore, P284 was identified as an important residue mediating the specific interaction between PTCR and 5α-DHT, and phylogenetic analysis showed that P284 is an evolutionarily conserved residue among animal carbonyl reductases, which suggests that the C-terminal tails of these reductases may have evolved under evolutionary pressure to increase the substrate specificity for C(21)-steroids and facilitate metabolic adaptation. Altogether, our RNA-Seq revealed that selective nsSNVs were associated with meat quality traits that could be useful for successful marker-assisted selection in pigs and also represents a useful resource to enhance understanding of protein folding, substrate specificity, and the evolution of enzymes such as carbonyl reductase.