• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.956-965
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• 2019

Objective: The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts. Methods: To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured. Results: SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts. Conclusion: Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.

• Animal Bioscience
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• v.34 no.1
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• pp.154-160
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• 2021

Objective: The effect of environmental enrichment on the behaviour of piglets in the nursery phase was evaluated. Methods: A total of 450 hybrid pigs (21 day old), including both females and uncastrated males, weighing approximately 6 kg, were distributed in a completely randomised design with 3 treatments and 3 replicates of 50 animals each. The treatments were: i) pen without environmental enrichment (control), ii) treatment consisting of continuous environmental enrichment (CEE) with rubber balls throughout the experimental period, and iii) treatment consisting of environmental enrichment with washed balls (EEWB) during the whole experimental period which were removed daily for washing. For the behavioural evaluation, 10 animals were randomly selected per replicate. The behavioural assessments were performed once a week, from 8 am to 6 pm, using images captured with a video camera. The data were submitted to non-parametric analyses, the means were compared using the Bonferroni test, and Person's correlations were also calculated. Results: A statistical difference (p = 0.001) was observed in the B5 (playful) behaviour; the animals in the EEWB treatment group had a higher frequency of this behaviour than animals in the control treatment group. The animals in the control group showed a higher frequency of B7 behaviour (lying down) (p = 0.026) than those in the EEWB and CEE treatment groups. The animals in the control group had a higher frequency of the B9 (belly nosing) behaviour than those in the EEWB group (p = 0.015). There was a tendency towards a higher frequency of behaviour B3 (walking in the pen) (p = 0.067) when the animals received the control treatment than treatments EEWB or CEE. Conclusion: The use of an enriching object improved the animal welfare and altered the correlation between the evaluated behaviours compared to the animals that did not have environmental enrichment.

• Animal Bioscience
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• v.34 no.4
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• pp.701-713
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• 2021

Objective: The present work was undertaken to evaluate the effects of storage time, choline chloride, and high concentrations of Cu and Zn on the kinetic behavior of vitamin degradation during storage in two vitamin premixes and four vitamin-trace mineral (VTM) premixes. Methods: Two vitamin premixes (with or without 160,000 mg/kg of choline) were stored at 25℃ and 60% humidity. Besides, four VTM premixes were used to evaluate the effects of choline (0 vs 40,000 mg/kg) and trace minerals (low CuSO4+ZnO vs high CuSO4+ZnO) on vitamin stability in VTM premixes stored in room, and the VTM premixes were stored in room temperature at 22℃. Subsamples from each vitamin and VTM premix were collected at 0, 1, 2, 3, 6, and 12 months. The retention of vitamin A (VA), vitamin D3 (VD3), vitamin E (VE), vitamin K3 (VK3), vitamin B1 (VB1), vitamin B2 (VB2), vitamin B3 (VB3), vitamin B5 (VB5), and vitamin B6 (VB6) in vitamin premixes and VTM premixes during storage was determined. The stability of vitamins in vitamin premixes and VTM premixes was determined and reported as the residual vitamin activity (% of initial) at each sampling point. Results: The effect of choline on VK3 retention was significant in vitamin premixes (p<0.05). The negative effect of storage time was significant for the retentions of VD3, VK3, VB1, VB2, VB5, and VB6 in vitamin premix (p<0.05). For VTM premixes, negative effect of storage time was significant (p<0.05) for the losses of vitamin in VTM premixes. Choline and high concentrations of Cu and Zn significantly increased VA, VK3, VB1, and VB2 loss during storage (p<0.05). The supplementation of high concentrations of Cu and Zn significantly decreased the concentrations of VD3 and VB6 (p<0.05) in VTM premixes at extended storage time. Conclusion: The maximum vitamin stability was detected in vitamin and VTM premixes containing no choline or excess Cu and Zn. The results indicated that extended storage time increased degradation of vitamin in vitamin or VTM premixes. These results may provide useful information for vitamin and VTM premixes to improve the knowledge of vitamin in terms of its stability.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.319-331
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• 2021

Heat stress (HS) causes adverse impacts on pig production and health. A potential biomarker of HS is required to predict its occurrence and thereby better manage pigs under HS. Information about the saliva metabolome in heat-stressed pigs is limited. Therefore, this study was aimed to investigate the effects of acute HS on the saliva metabolome and identify metabolites that could be used as potential biomarkers. Growing pigs (n = 6, 3 boars, and 3 gilts) were raised in a thermal neutral (TN; 25℃) environment for a 5-d adaptation period (CON). After adaptation, the pigs were first exposed to HS (30℃; HS30) and then exposed to higher HS (33℃; HS33) for 24 h. Saliva was collected after adaptation, first HS, and second HS, respectively, for metabolomic analysis using 1H-nuclear magnetic resonance spectroscopy. Four metabolites had significantly variable importance in the projection (VIP > 1; p < 0.05) different levels in TN compared to HS groups from all genders (boars and gilts). However, sex-specific characteristics affected metabolites (glutamate and leucine) by showing the opposite results, indicating that HS was less severe in females than in males. A decrease in creatine levels in males and an increase in creatine phosphate levels in females would have contributed to a protective effect from protein degradation by muscle damage. The results showed that HS led to an alteration in metabolites related to energy and protein. Protection from muscle damage may be attributed to the alteration in protein-related metabolites. However, energy-related metabolites showed opposing results according to sex-specific characteristics, such as sex hormone levels and subcutaneous fat layer. This study had shown that saliva samples could be used as a noninvasive method to evaluate heat-stressed pigs. And the results in this study could be contributed to the development of a diagnostic tool as a noninvasive biomarker for managing heat-stressed pigs.

• Journal of Life Science
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• v.31 no.8
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• pp.755-760
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• 2021

Swine diarrhea is a livestock disease that causes huge economic losses to pig farms. In general, diarrhea occurs because of the proliferation of pathogenic Escherichia coli (E. coli). The toxins produced by the proliferated E. coli cause edema in pigs. Although the proliferation of these coliforms can be prevented by using a vaccine, the vaccines containing chemically produced dead bacteria are not very effective, making it difficult to control the proliferation of E. coli. Therefore, there is a need to develop new, more effective vaccines. In this study, we prepared killed F4+ and F18ab+ E. coli, which induce diarrhea and edema in pigs, using the extracts of immune-induced silkworms containing antimicrobial peptides and examined their availability as a killed-bacteria vaccine. First, the antimicrobial activity analysis of the prepared immune-induced silkworm extract was conducted using the radial diffusion assay. The results showed high activity against both F4+ and F18ab+ E. coli. The production efficiency of E. coli dead cells was determined using the colony-counting method. The concentration of the E. coli dead cells was the highest (50 mg/ml) when treated at 4℃. In addition, the analysis of the prepared dead cells using a transmission electron microscope confirmed that E. coli leaked out of the cytoplasm and the cell membrane remained intact. Therefore, F4+ and F18ab+ E. coli produced using immune-induced silkworms extract are considered to be highly available as bacterial ghost vaccines that can help prevent swine diarrhea and the resulting edema.

• Animal Bioscience
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• v.35 no.8
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• pp.1235-1249
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• 2022

Objective: The purpose of this study was to evaluate the protection of glutamate (GLU) against the impairment in intestinal barrier function induced by lipopolysaccharide (LPS) stress in weaned pigs. Methods: Twenty-four weaned pigs were divided into four treatments containing: i) non-challenged control, ii) LPS-challenged control, iii) LPS+1.0% GLU, and iv) LPS+2.0% GLU. On day 28, pigs were treated with LPS or saline. Blood samples were collected at 0, 2, and 4 h post-injection. After blood samples collection at 4 h, all pigs were slaughtered, and spleen, mesenteric lymph nodes, liver and intestinal samples were obtained. Results: Dietary GLU supplementation inhibited the LPS-induced oxidative stress in pigs, as demonstrated by reduced malondialdehyde level and increased glutathione level in jejunum. Diets supplemented with GLU enhanced villus height, villus height/crypt depth and claudin-1 expression, attenuated intestinal histology and ultrastructure impairment induced by LPS. Moreover, GLU supplementation reversed intestinal intraepithelial lymphocyte number decrease and mast cell number increase induced by LPS stress. GLU reduced serum cortisol concentration at 4 h after LPS stress and downregulated the mRNA expression of intestinal corticotropin-releasing factor signal (corticotrophin-releasing factor [CRF], CRF receptor 1 [CRFR1], glucocorticoid receptor, tryptase, nerve growth factor, tyrosine kinase receptor A), and prevented mast cell activation. GLU upregulated the mRNA expression of intestinal transforming growth factor β. Conclusion: These findings indicate that GLU attenuates LPS-induced intestinal mucosal barrier injury, which is associated with modulating CRF signaling pathway.

• Animal Bioscience
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• v.35 no.11
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• pp.1752-1759
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• 2022

Objective: Effects of direct-fed Enterococcus faecium plus bacteriophages (EF-BP) were investigated as potential substitutes for pharmacological ZnO for weanling pigs. Methods: Dietary treatments were supplementations to a basal diet with none (NC), 3,000-ppm ZnO (PC), 1×10¹⁰ colony-forming units of E. faecium plus 1×10⁸ plaque-forming units (PFU) of anti-Salmonella typhimurium bacteriophages (ST) or 1×10⁶ PFU of each of anti-enterotoxigenic Escherichia coli K88 (F4)-, K99 (F5)-, and F18-type bacteriophages (EC) per kg diet. In Exp 1, twenty-eight 21-day-old crossbred weanling pigs were individually fed one of the experimental diets for 14 days and euthanized for histological examination on intestinal mucosal morphology. In Exp 2, 128 crossbred weanling pigs aged 24 days were group-fed the same experimental diets in 16 pens of 8 piglets on a farm with a high incidence of post-weaning diarrhea. Results: None of the diarrheal score or fecal consistency score (FCS), average daily gain (ADG), gain: feed ratio, structural variables of the intestinal villus, and goblet cell density, differed between the EF-BP (ST+EC) and NC groups, between EF-BP and PC, or between ST and EC, with the exception of greater gain: feed for EF-BP than for PC (p<0.05) during days 7 to 14 (Exp 1). In Exp 2, ADG was less for EF-BP vs PC during days 0 to 7 and greater for EF-BP vs NC during days 7 to 14. FCS peaked on day 7 and declined by day 14. Moreover, FCS was less for EF-BP vs NC, did not differ between EF-BP and PC, and tended to be greater for ST vs EC (p = 0.099). Collectively, EF-BP was comparable to or slightly less effective than PC in alleviating diarrhea and growth check of the weanling pigs, with ST almost as effective as PC, when they were group-fed. Conclusion: The E. faecium-bacteriophage recipe, especially E. faecium-anti-S. typhimurium, is promising as a potential substitute for pharmacological ZnO.

• Animal Bioscience
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• v.36 no.1
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• pp.29-42
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• 2023

Objective: Pigs, an ideal biomedical model for human diseases, suffer from about 50% early embryonic and fetal death, a major cause of fertility loss worldwide. However, identifying the causal variant remains a huge challenge. This study aimed to detect single nucleotide polymorphisms (SNPs) and candidate genes for the number of mummified (NM) piglets using the imputed whole-genome sequence (WGS) and validate the potential candidate genes. Methods: The imputed WGS was introduced from genotyping-by-sequencing (GBS) using a multi-breed reference population. We performed genome-wide association studies (GWAS) for NM piglets at birth from a Landrace pig populatiGWAS peak located on SSC11: 0.10 to 7.11 Mbp (Top SNP, SSC11:1,889,658 bp; p = 9.98E-13) was identified in cyclin dependent kinase on. A total of 300 Landrace pigs were genotyped by GBS. The whole-genome variants were imputed, and 4,252,858 SNPs were obtained. Various molecular experiments were conducted to determine how the genes affected NM in pigs. Results: A strong GWAS peak located on SSC11: 0.10 to 7.11 Mbp (Top SNP, SSC11:1,889,658 bp; p = 9.98E-13) was identified in cyclin dependent kinase 8 (CDK8) gene, which plays a crucial role in embryonic retardation and lethality. Based on the molecular experiments, we found that Y-box binding protein 1 (YBX1) was a crucial transcription factor for CDK8, which mediated the effect of CDK8 in the proliferation of porcine ovarian granulosa cells via transforming growth factor beta/small mother against decapentaplegic signaling pathway, and, as a consequence, affected embryo quality, indicating that this pathway may be contributing to mummified fetal in pigs. Conclusion: A powerful imputation-based association study was performed to identify genes associated with NM in pigs. CDK8 was suggested as a functional gene for the proliferation of porcine ovarian granulosa cells, but further studies are required to determine causative mutations and the effect of loci on NM in pigs.

• Animal Bioscience
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• v.35 no.5
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• pp.730-739
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• 2022

Objective: This study investigated the effects of dietary n-6:n-3 polyunsaturated fatty acid (PUFA) ratio on growth performance, blood indexes, tissue fatty acid composition and the gene expression in finishing pigs. Methods: Seventy-two crossbred ([Duroc×Landrace]×Yorkshire) barrows (68.5±1.8 kg) were fed one of four isoenergetic and isonitrogenous diets with n-6:n-3 PUFA ratios of 2:1, 3:1, 5:1, and 8:1. Results: Average daily gain, average daily feed intake and gain-to-feed ratio had quadratic responses but the measurements were increased and then decreased (quadratic, p<0.05). The concentrations of serum triglyceride, total cholesterol and interleukin 6 were linearly increased (p<0.05) with increasing of dietary n-6:n-3 PUFA ratio, while that of high-density lipoprotein cholesterol tended to decrease (p = 0.062), and high-density lipoprotein cholesterol:low-density lipoprotein cholesterol ratio and leptin concentration were linearly decreased (p<0.05). The concentration of serum adiponectin had a quadratic response but the measurement was decreased and then increased (quadratic, p<0.05). The proportion of C18:3n-3 was linearly decreased (p<0.05) in the longissimus thoracis (LT) and subcutaneous adipose tissue (SCAT) as dietary n-6:n-3 PUFA ratio increasing, while the proportion of C18:2n-6 and n-6:n-3 PUFA ratio were linearly increased (p<0.05). In addition, the expression levels of peroxisome proliferator-activated receptor gamma (PPARγ) and lipoprotein lipase in the LT and SCAT, and adipocyte fatty acid binding protein and hormone-sensitive lipase (HSL) in the SCAT had quadratic responses but the measurements were increased and then decreased (quadratic, p<0.05). The expression of HSL in the LT was linearly decreased (p<0.05) with increasing of dietary n-6:n-3 PUFA ratio. Conclusion: Dietary n-6:n-3 PUFA ratio could regulate lipid and fatty acid metabolism in blood and tissue. Reducing dietary n-6:n-3 PUFA ratio (3:1) could appropriately suppress expression of related genes in PPARγ signaling, and result in improved growth performance and n-3 PUFA deposition in muscle and adipose tissue in finishing pigs.

• Animal Bioscience
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• v.35 no.12
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• pp.1948-1956
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• 2022

Objective: This study was conducted to evaluate the effects of selenium (Se) source and level on the physiological response, reproductive performance, serum Se level, and milk composition in gestating sows. Methods: A total of 54 multiparous sows (Yorkshire×Landrace) with average body weight (BW), backfat thickness (BF), and parity were assigned to one of five treatments with 10 or 11 sows per treatment using a 2×2 factorial arrangement with one additional treatment in a completely randomized design. Inorganic or organic Se (IS or OS) sources were added to the diet at 0.30 ppm and 0.50 ppm Se. A non-Se-fortified corn-soybean meal basal diet served as a negative control. Treatments were as follows: i) Control: corn-soybean based diet, ii) IS30: control+inorganic Se 0.30 ppm, iii) IS50: control+inorganic Se 0.50 ppm, iv) OS30: control+ organic Se ppm, and v) OS50: control+organic Se 0.50 ppm. Results: At day 21 of lactation, piglet weight and weight gain in the OS treatments were higher than those in the IS treatments (p<0.05). Meanwhile, adding 0.5 ppm Se also resulted in the same significant differences in piglet BW and weight gain (p<0.05). Colostrum and milk Se concentrations increased (p<0.05) with Se level for both Se sources but were greater when sows were fed organic Se (p<0.05). Except for 24 hours postpartum, the Se concentrations were higher when sows were fed organic Se (p<0.05). Sow serum Se content was greater as Se levels increased from 0.3 ppm to 0.5 ppm at day 110 of gestation, 24 hours postpartum and day 21 of lactation (p<0.05). The pig serum Se concentration increased as the dietary Se level increased (p<0.05) and was higher when the sow dietary Se source was organic (p<0.05). Organic Se 0.5 ppm also had the highest serum Se level at two measured points (p<0.05). Conclusion: Consequently, supplementation with organic Se or 0.5 ppm Se in a gestating diet could improve piglet performance, the Se status of sows and piglets and milk composition, but organic Se at 0.5 ppm is optimal.