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β-Xylosidase and β-mannosidase in combination improved growth performance and altered microbial profiles in weanling pigs fed a corn-soybean meal-based diet

  • Liu, Shaoshuai;Ma, Chang;Liu, Ling;Ning, Dong;Liu, Yajing;Dong, Bing
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1734-1744
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    • 2019
  • Objective: In this study, two glycosidases (XMosidases), ${\beta}$-xylosidase and ${\beta}$-mannosidase, were investigated on their in vitro hydrolysis activities of feed and on the improvement of growth performance in vivo in weanling pigs. Methods: Enzyme activities of XMosidases in vitro were evaluated in test tubes and simulation of gastric and small intestinal digestion, respectively, in the presence of NSPase. In vivo study was performed in 108 weaned piglets in a 28-d treatment. Pigs were allotted to one of three dietary treatments with six replicate pens in each treatment. The three treatment groups were as follows: i) Control (basal diet); ii) CE (basal diets+CE); iii) CE-Xmosidases (basal diets+ CE+${\beta}$-xylosidase at 800 U/kg and ${\beta}$-mannosidase at 40 U/kg). CE was complex enzymes (amylase, protease, xylanase, and mannanase). Results: In vitro XMosidases displayed significant activities on hydrolysis of corn and soybean meal in the presence of non-starch polysaccharide degrading enzymes (xylanase and ${\beta}$-mannanase). In vitro simulation of gastric and small intestinal digestion by XMosidases showed XMosidases achieved $67.89%{\pm}0.22%$ of dry matter digestibility and $63.12%{\pm}0.21%$ of energy digestibility at $40^{\circ}C$ for 5 hrs. In weanling pigs, additional XMosidases to CE in feed improved average daily gain, feed conversion rate (p<0.05), and apparent total tract digestibility of crude protein (p = 0.01) and dry matter (p = 0.02). XMosidases also altered the gut bacterial diversity and composition by increasing the proportion of beneficial bacteria. Conclusion: Addition of a complex enzyme supplementation (contained xylanase, ${\beta}$-mannanase, protease and amylase), XMosidases (${\beta}$-xylosidase and ${\beta}$-mannosidase) can further improve the growth performance and nutrient digestion of young pigs.

ssc-miR-185 targets cell division cycle 42 and promotes the proliferation of intestinal porcine epithelial cell

  • Wang, Wei;Wang, Pengfei;Xie, Kaihui;Luo, Ruirui;Gao, Xiaoli;Yan, Zunqiang;Huang, Xiaoyu;Yang, Qiaoli;Gun, Shuangbao
    • Animal Bioscience
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    • v.34 no.5
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    • pp.801-810
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    • 2021
  • Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

Effects of dietary energy and protein levels on reproductive performance in gestating sows and growth of their progeny

  • Fang, Lin Hu;Jin, Ying Hai;Jeong, Jae Hark;Hong, Jin Su;Chung, Woo Lim;Kim, Yoo Yong
    • Journal of Animal Science and Technology
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    • v.61 no.3
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    • pp.154-162
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    • 2019
  • This experiment was conducted to evaluate the effect of dietary energy and crude protein (CP) levels on reproductive performance, litter performance, milk quality, and blood profiles in gestating sows. A total of 59 multiparous sows (Yorkshire ${\times}$ Landrace) with similar body weights (BW), backfat thickness (BF), and parity were assigned to one of six treatments with 9 or 10 sows per treatment using a $2{\times}3$ factorial arrangement and completely randomized design. The first factor was two levels of dietary metabolizable energy (ME) density (13.40 or 13.82 MJ/kg) and the second factor was three dietary protein levels based from 35 day in gestating phases (10.5%, 12%, and 13.5%). Backfat thickness change in lactating sows decreased linearly as CP level increased (p = 0.03). Increased energy level in the gestating sow diet tended to increase the total number of piglets born (p = 0.07), but piglet weight decreased (p = 0.02). Dietary CP level had a negative effect on colostrum quality. Casein, protein, total solid, and solids-not-fat concentrations decreased linearly and lactose level increased linearly as CP level in the gestating sow diet increased (casein%: p = 0.03; protein%: p = 0.04; lactose%: p = 0.06; total solids: p = 0.03; solid-not-fat: p = 0.03, respectively). However, improving ME by 0.42 MJ/kg had no significant effect on the chemical composition of sow colostrum. There were no significant differences in blood glucose concentration in gestating sows when sows were fed different levels of energy during gestation, but blood glucose increased at 21 day of lactation when energy increased by 0.42 MJ/kg (p = 0.04). Blood urea nitrogen concentration increased linearly when dietary CP levels increased at 110 day in gestation, 24-hours postpartum, and 21 days of lactation (linear, p < 0.05, p < 0.05, and p < 0.05, respectively), and it also increased when dietary energy increased at 110 days of gestation and 24-hours postpartum (p < 0.01, and p < 0.01, respectively). A gestating sow diet containing 13.82 MJ/kg ME and 10.5% CP can improve reproductive performance, litter performance, and colostrum quality.

Generation of a transgenic pig expressing human dipeptidylpeptidase-4 (DPP-4) (Human dipeptidylpeptidase-4(DPP-4) 발현 형질전환 돼지의 생산)

  • Chung, Hak Jae;Sa, Soo Jin;Baek, Sun Young;Cho, Eun Suek;Kim, Young Shin;Hong, Jun Ki;Cho, Kyu Ho;Kim, Ji Youn;Park, Mi Ryung;Kim, Kyung Woon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.9
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    • pp.306-314
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    • 2019
  • As dipeptidyl peptidase-4(DPP-4) inhibitors are used widely as a secondary treatment for type 2 diabetes because they tend to be well tolerated with minimal side effects, the human DPP-4(hDPP-4) gene was injected into a pig zygote through micro-injection, and 1-cell stage fertilized embryos were then transplanted surgically into the oviduct. Three pigs were fertilized with hDPP-4 genes and produced sixteen piglets, in which one male piglet was identified to be transgenic. Finally, transgenic pigs showing hDPP-4 gene expression in the tail were produced. Western blot and RT-PCR analysis confirmed that the hDPP-4 is expressed strongly in the membrane cells of the transgenic pig, and that the hDPP-4 gene appears in various tissues and tails. This suggests that the expression vector is normally expressed in transgenic pigs. These results are anticipated to be a model animal to check the endocrine function for insulin resistance that occurs in a hDPP-4 transgenic pig and to increase its value for use as a material in newly developed medicines.

Determination of dietary Ca and P levels and their equivalence values of phytase and vitamin D3 for improved growth performance in weanling pigs

  • Ogola, Oketch Elijah;Choi, Jun Seung;Hong, Jun Seon;Kim, Yu Bin;Nawarathne, Shan Randima;Yu, Myunghwan;Heo, Jung Min
    • Korean Journal of Agricultural Science
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    • v.48 no.3
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    • pp.397-412
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    • 2021
  • In Experiment 1, the impact of using diets sufficient in P on the performance of weaned piglet growth with or without a super dose of phytase was investigated. One hundred and twelve piglets were randomized into four treatments with 7 replicates (4 piglets·cage-1). Bodyweight (BW), daily gain (ADG), daily intake (ADFI) and feed conversion ratios (FCR) were measured over three weeks. Treatments included a positive control (PC) with a Ca : P ratio of 0.80 : 0.50%. and the negative control (NC) had a Ca : P ratio of 0.60 : 0.50%. The PC plus phytase was denoted as PC+, and the NC with the phytase diet was designated as NC+. The PC diet was formulated to meet or exceed the NRC (2012) standards for pigs weighing 7 - 25 kgs. Better results were obtained with the PC diet (p > 0.1) compared to the NC diet. The super dose of phytase in the PC+ diet significantly improved the overall FCR (p < 0.1). For Experiment 2, the impact of different calcium (Ca)/phosphorous (P) ratios with or without supplementation of vitamin D3 was determined. Ninety-six piglets were randomized into four treatments with 6 replicates (4 piglets·cage-1). The PC, NC1 and NC2 diets had ratios of 0.80 : 0.50, 0.80 : 0.45, and 0.60 : 0.45%, respectively. The fourth diet (PCV) was the PC diet fortified with vitamin D3. Improved results were observed with the PC diet (p > 0.1) compared to the NC diets. Supplementing dietary 25-OH-D3 in the PCV diet led to numerically higher ADG and ADFI scores with lower FCR values at the early nursery stage (p = 0.0044), but the vitamin supplement did not significantly influence the growth.

Effects of dietary supplementation of glucose oxidase, catalase, or both on reproductive performance, oxidative stress, fecal microflora and apoptosis in multiparous sows

  • Sun, Xiaojiao;Piao, Longguo;Jin, Haifeng;Nogoy, K. Margarette C.;Zhang, Junfang;Sun, Bin;Jin, Yi;Lee, Dong Hoon;Choi, Seong-Ho;Smith, Stephen B;Li, Xiangzi
    • Animal Bioscience
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    • v.35 no.1
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    • pp.75-86
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    • 2022
  • Objective: The objective of this experiment was to investigate the effect of dietary glucose oxidase (GOD), catalase (CAT), or both supplementation on reproductive performance, oxidative stress, and apoptosis in sows. Methods: A total of 104 multiparous sows were randomly assigned to four groups (n = 26) with each group given a basal diet, basal diet plus GOD at 60 U/kg, basal diet plus CAT at 75 U/kg, and basal diet plus GOD at 60 U/kg and CAT at 75 U/kg. Sows were fed the experimental diets throughout gestation and lactation. Results: Dietary GOD supplementation increased average daily feed intake of sows and litter weight at weaning (p<0.05). Dietary CAT supplementation reduced the duration of parturition, stillbirth, and piglet mortality and increased growth performance of weaned piglets (p<0.05). Dietary GOD and CAT supplementation enhanced antioxidant enzyme activities and lessened oxidative stress product levels in plasma of sows and elevated antioxidant capacity of 14-day milk and plasma in weaned piglets (p<0.05). Dietary GOD supplementation increased fecal Lactobacillus counts and reduced Escherichia coli counts of sows (p<0.05). Compared with the basal diet, the GOD diet reduced fecal Escherichia coli counts of sows, but the addition of CAT did not reduce Escherichia coli counts in the GOD diet. Dietary GOD and CAT supplementation reduced the apoptosis rate of the liver, endometrium, and ovarian granulosa cells in sows (p<0.05). In the liver, uterus, and ovary of sows, the mRNA expression of caspase-3 and caspase-9 was downregulated by dietary GOD and CAT supplementation (p<0.05). Conclusion: Dietary GOD and CAT supplementation could improve the antioxidant capacity of sows and weaned piglets, and alleviate hepatic, ovarian and uterine apoptosis by weakening apoptosis-related gene expression. Glucose oxidase regulated fecal microflora of sows, but supplementation of CAT to GOD could weaken the inhibitory effect of GOD on fecal Escherichia coli.

Effects of protein content and the inclusion of protein sources with different amino acid release dynamics on the nitrogen utilization of weaned piglets

  • Hu, Nianzhi;Shen, Zhiwen;Pan, Li;Qin, Guixin;Zhao, Yuan;Bao, Nan
    • Animal Bioscience
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    • v.35 no.2
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    • pp.260-271
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    • 2022
  • Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

A regression for estimating metabolizable glucose in diets of weaned piglets for optimal growth performance

  • Lv, Liangkang;Feng, Zhi;Zhang, Dandan;Lei, Long;Zhang, Hui;Liu, Zhengya;Ren, Ying;Zhao, Shengjun
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1643-1652
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    • 2021
  • Objective: Two experiments were conducted to provide a new approach for evaluating feed nutritional value by metabolizable glucose (MG) in piglet diets with different levels of starch and crude fiber. In Exp 1, a regression equation for MG was generated. In Exp 2, the equation was verified, and the optimal growth performance of piglets under appropriate MG levels was tested. Methods: In Exp 1, 20 weaned piglets (7.74±0.81 kg body weight [BW]) were randomly assigned to 1 of 4 treatments, including the basal diet containing different levels of MG (starch, 25.80%, 31.67%, 45.71%, 49.36%; crude fiber, 1.23%, 1.35%, 1.80%, 1.51%). The piglets were implanted with an ileal fistula, cannulation of the carotid artery, portal vein, and mesenteric artery. The chyme from the ileum fistula and blood samples were collected. In Exp 2, 30 weaned piglets (8.96±0.50 kg BW) were randomly assigned to 1 of 5 treatments, including the experimental diets with different levels of MG (37.6, 132.5, 300.0, 354.3, and 412.5 g/kg). The piglets' BW, and feed consumption were recorded to calculate growth performance during the 28-d experiment. Results: In Exp 1, the MG levels in 4 diets were 239.62, 280.68, 400.79, and 454.35 g/kg. The regression equation for the MG levels and dietary nutrients was: Y (MG) = 12.13×X1 (starch)+23.18×X2 (crude fiber)-196.44 (R2 = 0.9989, p = 0.033). In Exp 2, treatments with 132.5 and 300.0 g/kg MG significantly (p<0.05) increased average daily gain and feed conversion efficiency of weaned piglets, increased digestibility of crude fat, and had no effect on digestibility of crude protein compared to 300.0 to 412.5 g/kg MG. Conclusion: The pig model combining the ileum fistula and cannulation of blood vessels was successfully used to determine the dietary MG levels. The recommended MG level in weaned pig diets is 132.5 to 300.0 g/kg.

Effect of Saccharomyces cerevisiae boulardii on sows' farrowing duration and reproductive performance, and weanling piglets' performance and IgG concentration

  • Sun, Haiqing;de Laguna, Fernando Bravo;Wang, Shuai;Liu, Fengju;Shi, Liang;Jian, Haidi;Hu, Xiaoxia;Qin, Peng;Tan, Jiajian
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.10-22
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    • 2022
  • We studied the effects of Saccharomyces cerevisiae boulardii CNCM I-1079 (LSB) supplemented to lactating sows on reproductive traits and farrowing duration and to piglets from day 7 of life on post-weaning performance and IgG concentration. Ninety-six Landrace × Yorkshire sows started the trial 5 days before the expected farrowing date. Sows were distributed into 2 groups according to parity number and backfat thickness: control (CON: regular lactation diet) and LSB (CON + LSB at 2 × 109 colony forming units [CFU]/kg of feed). Seven days after birth, litters were randomly selected from each group and supplemented creep feed with or without LSB at 2 × 109 CFU/kg. At weaning, piglets from CON sows were shifted to a commercial farm and allocated to 14 pens in groups of 25 piglets/pen according to the creep feed supplemented during lactation. Piglets followed a 3-phase feeding program: creep, pre-starter and starter, with or without LSB at 2 × 109 CFU/kg LSB in creep and pre-starter, and 1 × 109 CFU/kg LSB in starter. The piglets were vaccinated against classical swine fever on days 41 and 72 of life. One day before each vaccination and at the end of the trial, blood samples were collected from 15 randomly selected piglets per treatment and assessed for total IgG. Supplemented sows with non-supplemented litters displayed the lowest backfat thickness loss during lactation (p < 0.05). The LSB supplementation shortened farrowing duration (p < 0.05) and increased feed intake (p < 0.05) during the first week of lactation. The LSB-fed piglets were heavier at the end of creep (p < 0.05), pre-starter (p < 0.05), and the trial (p < 0.05); grew faster during creep (p < 0.05), starter (p < 0.05), and overall (p < 0.05); and displayed an improved feed conversion ratio during creep (p < 0.05). Total IgG content was higher at days 40 (p < 0.05) and 71 (p < 0.05) in LSB-fed piglets. We conclude that supplementing sows with Saccharomyces cerevisiae boulardii CNCM I-1079 from late gestation until weaning shortens farrowing duration, increases feed intake, and minimizes backfat losses during lactation. When supplemented to piglet diet, post-weaning performance is improved. This improvement observed could be linked to a better immune status, as suggested by the higher IgG.

Establishment of Hysterectomy for Gnotobiotic Pig Production (무균돼지 생산을 위한 자궁적출술 확립)

  • Nho, W.G.;Lee, J.H.;Kim, W.Y.;Yeo, J.M.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.10 no.1
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    • pp.91-99
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    • 2008
  • Gnotobiotic piglets were routinely produced by hysterectomy. In this study, 22 pregnant miniature pigs (111th to 113th day of gestation) were used for hysterectomy. Before surgery, 14 pigs were insensibilizated by Ketamine 50® plus CO2 gas and 8 pigs by a slaughter pistol. The high level of Ketamine 50® (0.09㎖/kg) was faster (146 vs 283 seconds) in surgery but the time taken for complete revival of one piglet was more prolonged (427 vs 64 seconds) than 0.03㎖/kg level. In hysterectomies with a slaughter pistol, surgery time was faster (470 vs 155 seconds) and the rate of alive piglets was higher (97.0 vs 83.8%) than in those with Ketamine 50®. There were no problems in the rate of alive newborn piglets even when sows were hysterectomized at 3 days prepartum.