• Animal Bioscience
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• v.36 no.8
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• pp.1263-1273
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• 2023

Objective: This study was conducted to evaluate the effects of crude protein (CP) levels on the physiological response, reproductive performance, blood profiles, milk composition and odor emission in gestating sows. Methods: Seventy-two multiparous sows (Yorkshire×Landrace) of average body weight (BW), backfat thickness, and parity were assigned to one of six treatments with 10 or 11 sows per treatment in a completely randomized design. Experimental diets with different CP levels were as follows: i) CP11, corn-soybean-based diet containing 11% CP; ii) CP12, corn-soybean-based diet containing 12% CP; iii) CP13, corn-soybean-based diet containing 13% CP; iv) CP14, corn-soybean-based diet containing 14% CP; v) CP15, corn-soybean-based diet containing 15% CP; and vi) CP16: corn-soybean-based diet containing 16% CP. Results: There was no significant difference in the performance of sow or piglet growth when sows were fed different dietary protein levels. Milk fat (linear, p = 0.05) and total solids (linear, p = 0.04) decreased as dietary CP levels increased. Increasing dietary CP levels in the gestation diet caused a significant increase in creatinine at days 35 and 110 of gestation (linear, p = 0.01; linear, p = 0.01). The total protein in sows also increased as dietary CP levels increased during the gestation period and 24 hours postpartum (linear, p = 0.01; linear, p = 0.01). During the whole experimental period, an increase in urea in sows was observed when sows were fed increasing levels of dietary CP (linear, p = 0.01), and increasing blood urea nitrogen (BUN) concentrations were observed as well. In the blood parameters of piglets, there were linear improvements in creatinine (linear, p = 0.01), total protein (linear, p = 0.01), urea (linear, p = 0.01), and BUN (linear, p = 0.01) with increasing levels of dietary CP as measured 24 hours postpartum. At two measurement points (days 35 and 110) of gestation, the odor gas concentration, including amine, ammonia, and hydrogen sulfide, increased linearly when sows fed diets with increasing levels of dietary CP (linear, p = 0.01). Moreover, as dietary CP levels increased to 16%, the odor gas concentration was increased with a quadratic response (quadratic, p = 0.01). Conclusion: Reducing dietary CP levels from 16% to 11% in a gestating diet did not exert detrimental effects on sow body condition or piglet performance. Moreover, a low protein diet (11% CP) may improve dietary protein utilization and metabolism to reduce odor gas emissions in manure and urine in gestating sows.

• Animal Bioscience
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• v.36 no.6
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• pp.869-878
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• 2023

Objective: Hyperketonemia remains a major metabolic issue of serious milk production and a major health concern in early lactation cows. Oral supplementation of glucose precursors (GP) can be used to prevent hyperketonemia in dairy cows. The objective of this study was to compare the beneficial effects of orally supplementing a mixture of GP on metabolic health indicators and milk production status of primiparous (PP) and multiparous (MP) dairy cows. Methods: Twenty-eight Holstein cows were blocked by expected date of parturition, previous lactation yield, and parity. The cows were randomly allocated to one of the four treatment groups (n = 7 cows/group) based on their parity and GP supplementation: i) PP cows fed basal diet only (PP-CON), ii) PP cows with oral supplementation of GP (PP-GP), iii) MP cows fed basal diet only (MP-CON), and iv) MP cows with oral supplementation of GP (MP-GP). Glucose precursor (glycoline liquid) was orally drenched (300 mL/d) in GP cows from 7 days prepartum through 7 days postpartum. Other than GP supplementation, all cows were fed similar pre- and postpartum basal diets. Results: In both pre- and postpartum periods, serum glucose concentration was increased, whereas β-hydroxybutyrate and free fatty acids were decreased in GP cows compared with the CON cows. Milk yield and milk components were statistically not different between GP and CON cows over the first 9 week of lactation. The yield of actual milk, energy-corrected milk, 63-days cumulative milk, colostrum yield, and calf birth weight remained higher in MP cows compared with PP cows. Conclusion: Oral drenching of GP around calving can be recommended to successfully improve the metabolic health and reduce the negative effects of hyperketonemia not only in MP but also in PP dairy cows.

• Animal Bioscience
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• v.36 no.6
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

• Animal Bioscience
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• v.36 no.6
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• pp.899-907
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• 2023

Objective: The better understanding of laying pattern of birds is crucial for developing breed-specific proper breeding scheme and management. Methods: Daily egg production until 50 wk of age of six chicken breeds including one layer (White Leghorn, WL), three dual-purpose (Rhode Island Red, RIR; Columbian Plymouth Rock, CR; and Barred Plymouth Rock, BR), one synthetic dwarf (DY), and one indigenous (Beijing-You Chicken, BYC) were used to characterize their clutch traits and egg production. The age at first egg, egg number, average and maximum clutch length, pause length, and number of clutches and pauses were calculated accordingly. Results: The egg number and average clutch length in WL, RIR, CR, and BR were higher than those in DY and BYC (p<0.01). The numbers of clutches and pauses, and pause length in WL, RIR, CR, and BR were lower than those in DY and BYC (p<0.01). The coefficient variations of clutch length in WL, RIR, CR, and BR (57.66%, 66.49%, 64.22%, and 55.35%, respectively) were higher than DY (41.84%) and BYC (36.29%), while the coefficient variations of egg number in WL, RIR, CR, and BR (9.10%, 9.97%, 10.82%, and 9.92%) were lower than DY (15.84%) and BYC (16.85%). The clutch length was positively correlated with egg number (r = 0.51 to 0.66; p<0.01), but not correlated with age at first egg in all breeds. Conclusion: The six breeds showed significant different clutch and egg production traits. Due to the selection history, the high and median productive layer breeds had higher clutch length than those of the less productive indigenous BYC. The clutch length is a proper selection criterion for further progress in egg production. The age at first egg, which is independent of clutch traits, is especially encouraged to be improved by selection in the BYC breed.

• Animal Bioscience
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• v.36 no.6
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• pp.861-868
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• 2023

Objective: Loin muscle area (LMA) is an important target trait of pig breeding. This study aimed to identify single nucleotide polymorphisms (SNPs) and genes associated with LMA in the Duroc×(Landrace×Yorkshire) crossbred pigs (DLY). Methods: A genome-wide association study was performed using the Illumina 50K chip to map the genetic marker and genes associated with LMA in 511 DLY pigs (255 boars and 256 sows). Results: After quality control, we detected 35,426 SNPs, including six SNPs significantly associated with LMA in pigs, with MARC0094338 and ASGA0072817 being the two key SNPs responsible for 1.77% and 2.48% of the phenotypic variance of LMA, respectively. Based on previous research, we determined two candidate genes (growth hormone receptor [GHR] and 3-oxoacid Co A-transferase 1 [OXCT1]) that are associated with fat deposition and muscle growth and found further additional genes (MYOCD, ARHGAP44, ELAC2, MAP2K4, FBXO4, FBLL1, RARS1, SLIT3, and RANK3) that are presumed to have an effect on LMA. Conclusion: This study contributes to the identification of the mutation that underlies quantitative trait loci associated with LMA and to future pig breeding programs based on marker-assisted selection. Further studies are needed to elucidate the role of the identified candidate genes in the physiological processes involved in LMA regulation.

• Animal Bioscience
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• v.36 no.6
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• pp.943-952
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• 2023

Objective: For decades, LYD ([Landrace×Yorkshire] ♀×Duroc ♂) pigs are the most commonly-used commercial breed for meat production in Korea. Recently, due to the increasing demand for premium pork, the National Institute of Animal Science (Korea) has synthesized a novel pig breed named Woori Heukdon (WHD). This study aimed at comparing the carcass traits, meat yield and quality characteristics between the LYD and WHD pigs under identical rearing condition. Methods: The WHD and LYD pigs (n = 15 each) were reared under identical conditions and fed the same commercial diet until reaching recommended market weight (100 to 120 kg). After slaughter, the carcasses were evaluated for traits and meat yield, and the meat quality was assessed on shoulder butt and belly cuts. Results: Although no significant differences (p>0.05) occurred in slaughter weight between two pig types, WHD had a lower meat yield (by about 6 kg corresponding to approximately 7%) compared to the LYD pigs (p<0.05). The WHD had a higher fat content (by 4.26% and 13.52% in the shoulder butt and belly, respectively) compared to those of LYD pigs (p<0.05). The WHD meat showed a lower cooking loss and higher a* (redness) value (p<0.05). The WHD belly had a significantly (p<0.05) higher oleic acid content and concentrations of nonanal, octanal and decanal associated with fatty odor while, the LYD meat had a higher number of pyrazines associated with roasty odor. Regarding sensory quality, higher flavor, juiciness, and acceptability higher scores were given for the WHD meat than for the LYD meat (p<0.05). Conclusion: Under identical rearing conditions the WHD exhibited a better meat quality and sensory properties. However, the use of this diet resulted in a high fat level that may be associated with high trimming loss for the WHD.

• Animal Bioscience
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• v.36 no.6
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• pp.962-972
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• 2023

Objective: This study determined the effects of dietary treatments and castration on meat quality, fatty acids (FAs) profiles, and volatile compounds in Korean native black goats (KNBG, Capra hircus coreanae), including the relationship between the population of rumen microbiomes and meat FA profiles. Methods: Twenty-four KNBG (48.6±1.4 kg) were randomly allocated to one of four treatments arranged into a 2×2 factorial structure. The factors were dietary forage to concentrate ratio (high forage [HF, 80:20] and low forage [LF, 20:80]), and a castration treatment (castration [CA] vs non-castration [NCA]). Results: Among meat quality traits, the CA group exhibited a higher percentage of crude fat and water holding capacity (p<0.05). The profiles of the saturated fatty acid (SFA) in meat sample derived from CA KNBG showed a significantly lower percentage compared to NCA individuals, due to the lower proportion of C14:0 and C18:0. Feeding a high-forage diet to KNBG increased the formation of C18:1n7, C18:3n3, C20:1n9, C22:4n6 in meat, and polyunsaturated fatty acid (PUFA) profiles (p<0.05). Consequently, the n6:n3 ratio declined (p<0.05). There was an interaction between dietary treatment and castration for formation of C20:5n3 (p<0.05), while C18:1n9, C22:6n3, monounsaturated fatty acid (MUFA) and the MUFA:SFA ratio were influenced by both diet and castration (p<0.05). Nine volatile compounds were identified and were strongly influenced by both dietary treatments, castration (p<0.05), and their interaction. In addition, principal component analysis (PCA) revealed distinctly different odor patterns in the NCA goats fed LF diets. Spearman correlation analysis showed a high correlation between rumen bacteria and meat PUFAs. Conclusion: These results suggest the essential effects of the rumen microbial population for the synthesis of meat FAs and volatile compounds in KNBG meat, where dietary intake and castration also contribute substantially.

• Animal Bioscience
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• v.36 no.6
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• pp.929-942
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• 2023

Objective: This study was conducted to evaluate the effects of β-glucan with vitamin E supplementation on the growth performance, blood profiles, immune response, pork quality, pork flavor, and economic benefit in growing and finishing pigs. Methods: A total of 140 growing pigs ([Yorkshire×Landrace]×Duroc) were assigned to five treatments considering sex and initial body weight (BW) in 4 replications with 7 pigs per pen in a randomized complete block design. The experimental diets included a corn-soybean meal-based basal diet with or without 0.05% or 0.1% β-glucan and 0.02% vitamin E. The pigs were fed the diets for 12 weeks (phase I, 0 to 3; phase II, 3 to 6; phase III, 6 to 9; phase IV, 9 to 12). The BW and feed intake were measured at the end of each phase. Blood samples were collected at the end of each phase. Four pigs from each treatment were selected and slaughtered for meat quality. Economic benefit was calculated considering the total feed intake and feed price. Pork flavor was analyzed through inosine monophosphate analysis. Results: The average daily gain and feed efficiency were improved compared to the control when β-glucan or vitamin E was added. Supplementing 0.05% β-glucan significantly increased the lymphocyte concentration compared to the addition of 0.1% β-glucan and the content of vitamin E in the blood increased when 0.02% vitamin E was added. The treatment with 0.1% β-glucan and 0.02% vitamin E showed the most economic effect because it had the shortest days to market weight and the lowest total feed cost. The addition of β-glucan or vitamin E had a positive role in improving the flavor of pork when considering that the content of inosine monophosphate was increased. However, carcass traits and meat quality were not affected by β-glucan or vitamin E. Conclusion: The addition of 0.1% β-glucan with 0.02% vitamin E in growing and finishing pig diets showed great growth performance and economic effects by supplying vitamin E efficiently and by improving the health condition of pigs due to β-glucan.

• Animal Bioscience
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• v.36 no.10
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• pp.1488-1498
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• 2023

Objective: α_S1-Casein is more closely associated with milk allergic reaction than other milk protein components. microRNA (miRNA) is a class of small non-coding RNAs that modulate multiple biological progresses by the target gene. However, the post-transcriptional regulation of α_S1-casein expression by miRNA in ruminants remains unclear. This study aims to explore the regulatory roles of miR-380-3p on α_S1-casein synthesis in goat mammary epithelial cells (GMEC). Methods: α_S1-Casein gene and miR-380-3p expression was measured in dairy goat mammary gland by quantitative real-time polymerase chain reaction (qRT-PCR). miR-380-3p overexpression and knockdown were performed by miR-380-3p mimic or inhibitor in GMEC. The effect of miR-380-3p on α_S1-casein synthesis was detected by qRT-PCR, western blot, luciferase and chromatin immunoprecipitation assays in GMEC. Results: Compared with middle-lactation period, α_S1-casein gene expression is increased, while miR-380-3p expression is decreased during peak-lactation of dairy goats. miR-380-3p reduces α_S1-casein abundance by targeting the 3'-untranslated region (3'UTR) of α_S1-casein mRNA in GMEC. miR-380-3p enhances β-casein expression and signal transducer and activator of transcription 5a (STAT5a) activity. Moreover, miR-380-3p promotes β-casein abundance through target gene α_S1-casein, and activates β-casein transcription by enhancing the binding of STAT5 to β-casein gene promoter region. Conclusion: miR-380-3p decreases α_S1-casein expression and increases β-casein expression by targeting α_S1-casein in GMEC, which supplies a novel strategy for reducing milk allergic potential and building up milk quality in ruminants.

• Animal Bioscience
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• v.36 no.9
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• pp.1336-1349
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• 2023

Objective: The study was conducted to screen differentially expressed miRNAs in sows at early pregnancy by high-throughput sequencing and explore its mechanism of action on embryo implantation. Methods: The blood serum of pregnant and non-pregnant Landrace×Yorkshire sows were collected 14 days after artificial insemination, and exosomal miRNAs were purified for high throughput miRNA sequencing. The expression patterns of 10 differentially expressed (DE) miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR quantified the abundance of serum exosomal miR-192 in pregnant and control sows, and the diagnostic power was assessed by receiver operating characteristic (ROC) analysis. The target genes of DE miRNAs were predicted with bioinformatics software, and the functional and pathway enrichment analysis was performed on gene ontology and the Kyoto encyclopedia of genes and genomes terms. Furthermore, a luciferase reporter system was used to identify the target relation between miR-192 and integrin alpha 4 (ITGA4), a gene influencing embryo implantation in pigs. Finally, the expression levels of miRNAs and the target gene ITGA4 were analyzed by qRT-PCR, and western blot, with the proliferation of BeWo cells detected by cell counting kit-8 (CCK-8). Results: A total of 221 known miRNAs were detected in the libraries of the pregnant and non-pregnant sows, of which 55 were up-regulated and 67 were down-regulated in the pregnant individuals compared with the non-pregnant controls. From these, the expression patterns of 10 DE miRNAs were validated. The qRT-PCR analysis further confirmed a significantly higher expression of miR-192 in the serum exosomes extracted from pregnant sows, when compared to controls. The ROC analysis revealed that miR-192 provided excellent diagnostic accuracy for pregnancy (area under the ROC curve [AUC]=0.843; p>0.001). The dual-luciferase reporter assay indicated that miR-192 directly targeted ITGA4. The protein expression of ITGA4 was reduced in cells that overexpressed miR-192. Overexpression of miR-192 resulted in the decreased proliferation of BeWo cells and regulated the expression of cell cycle-related genes. Conclusion: Serum exosomal miR-192 could serve as a potential biomarker for early pregnancy in pigs. miR-192 targeted ITGA4 gene directly, and miR-192 can regulate cellular proliferation.