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The Effects of Alismatis Rhizoma Extract on Allergic Inflammation in RBL-2H3 Mast Cells and OVA/alum-Sensitized Mice (택사 (澤瀉, Alismatis Rhizoma) 추출물이 RBL-2H3 비만세포와 OVA/alum 감작 생쥐의 알레르기 염증 반응에 미치는 영향)

  • Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.32 no.3
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    • pp.1-15
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    • 2018
  • Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

Transfer of Porcine Embryos Injected with Sperm Carrying with Exogenous DNA

  • Cho, Seong-Keun;Cho, Hwang-Yun;Park, Mi-Ryung;Park, Jong-Sik;Yoo, Jae-Gyu;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.61-61
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    • 2001
  • The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.

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Anti-inflammatory Effect of the Robinia pseudoacacia L. High Temperature Extract (아까시 나무 고온추출물의 항염증 효과)

  • Nho, Jong Hyun;Kang, Byoung Man;Jung, Won Seok
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.294-302
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    • 2018
  • This study was conducted to compare anti-inflammatory effect of Robinia pseudoacacia L. using different extraction methods (water extraction, ethanol extraction and high temperature extraction). We investigated anti-inflammatory effect of Robinia pseudoacacia L. extract (RP1, water extract; RP2, ethanol extract; RP3, high temperature extract) on lipopolysaccharide (LPS)-stimulated inflammation using Raw 264.7 cell. Cells were treated with various concentrations (12.5, 25, 50, 100 or $200{\mu}g/m{\ell}$) of water extract, ethanol extract and high temperature extract. Cytotoxicity was not observed on Raw 264.7 cells, LPS-stimulated production of NO (nitric oxide), $PGE_2$ (prostaglandin $E_2$) and cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$) was reduced by RP3 treatment more than RP1 and RP2. In conclusion, these results indicated that inflammation on Raw 264.7 cells was improved by RP3. Treatment of RP3 could be used to natural medicine for improving inflammatory response. However, further experiment is required to observe how the high temperature extraction at $500^{\circ}C$ for 48 h influences on alteration of active ingredient in Robinia pseudoacacia L., and conducts the inflammation signal pathway on Raw 264.7 cells.

Antioxidative, Antimicrobial and Cytotoxic activities of Fagopyrum esculentum $M{\ddot{o}}ench$ Extract in Germinated Seeds (발아 메밀 추출물의 항산화.항균활성 및 세포독성)

  • Hwang, Eun-Ju;Lee, Sook-Young;Kwon, Su-Jung;Park, Min-Hee;Boo, Hee-Ock
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.1
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    • pp.1-7
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    • 2006
  • This research was conducted to investigate the possibilities of usage of germinated-buckwheat (Fagopyrum esculentum $M{\ddot{o}}ench$) by examining antioxidative, antimicrobial and cytotoxic effects of extracts from different germinated root length of buckwheat. Antioxidant activity $(RC_{50})$ was shown higher in extracts of non-germinated seed $(50.41\;{\mu}g/mL)$ and root length 10 mm $(80.57\;{\mu}g/mL)$, 2 mm $(93.77\;{\mu}g/mL)$, 5 mm $(107.09\;{\mu}g/mL)$ than BHT $(163.96\;{\mu}g/mL)$ as a synthetic antioxidant. In antimicrobial activity, non-germinated and germinated seeds were formed inhibitory zone against S. aureus $(4{\sim}10\;mm)$, P. aeruginosa $(2{\sim}9\;mm)$ at the concentrations of $10{\sim}40\;mg/mL$ but B. subtilis, E. coli and S. typhimurium were not apparent antimicrobial activity. Extracts of germinated seed also decreased their antimicrobial activity compared to non-germinated seed extract. In addition, the growth of Calu-6 was inhibited of both 5 mm root length germinated and non-germinated seeds $(800\;{\mu}g/mL)$ as 95.12% and 87.15%, respectively, but these did not show any influence on cytotoxic effect against MCF-7 and Caco-2 cell lines. Extracts of 2 mm and 5 mm germinated seeds were also inhibited against Calu-6 and SNU-601 cell lines.

Enterococcus faecium FC-K Derived from Kimchi Is a Probiotic Strain That Shows Anti-Allergic Activity

  • Rho, Man-Kwang;Kim, Young-Eun;Rho, Hyun-In;Kim, Tae-Rahk;Kim, Yoon-Bum;Sung, Won-Kyung;Kim, Tae-Woo;Kim, Dae-Ok;Kang, Hee
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1071-1077
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    • 2017
  • A rise in the occurrence of allergic diseases is attributed to the dysregulated balance of type 1/type 2 immunity, where type 2 T-helper (Th2) cells predominate over type 1 T-helper (Th1) cells, leading to an abnormally increased production of IgE in response to unharmful antigens. Kimchi, a traditional Korean fermented food, is a rich source of beneficial lactic acid bacteria. In this study, we investigated the ability of Enterococcus faecium FC-K derived from kimchi to induce type I immunity in the presence of Th2 polarizing conditions in vitro and in vivo. Stimulation of mouse peritoneal macrophages with E. faecium FC-K induced the production of tumor necrosis factor alpha, interleukin (IL)-6, and IL-12. Under the in vitro Th2 conditions in which splenic T cells were activated in the presence of IL-4, E. faecium FC-K enhanced the ability of T cells to produce interferon $(IFN)-{\gamma}$. Using the ovalbumin (OVA)-induced allergy model, male BALB/c mice receiving E. faecium FC-K reduced the serum level of total IgE, but not that of OVA-specific IgE. Furthermore, the population of activated splenic B cells during OVA immunization was decreased in E. faecium FC-K-treated mice, accounting for a reduction of total IgE in the serum. Restimulating splenocytes from OVA-immunized mice with OVA ex vivo resulted in an increased production of $IFN-{\gamma}$, with no effect on IL-4, in E. faecium FC-K-treated mice. These observations provide the evidence that E. faecium FC-K can be a beneficial probiotic strain that can modulate the Th2-mediated pathologic response.

Supplementation of Holstein Cows with Low Doses of Bovine Somatotropin (bST) Prepartum and Postpartum Affects Physiological Adaptations and Milk Production

  • Liboni, M.;Gulay, M.S.;Hayen, M.J.;Belloso, T.I.;Head, H.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.404-413
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    • 2008
  • Major objectives were to evaluate effects of three schemes of bST-supplementation of Holstein cows (142.8 mg/14 d, POSILAC) during the prepartum and/or postpartum periods through 63 d (${\pm}3d$) of lactation. Measures evaluated the potential of treatments to improve body weight (BW) and body condition score (BCS), provoke changes in plasma concentrations of somatotropin (ST) and IGF-I, and improve milk yield, milk composition (percentages of protein and fat, and somatic cell counts), and several calving variables. Multiparous Holstein cows were randomly assigned to a $2{\times}2$ factorial arrangement of treatments (TRT) to give four groups (I = no bST, n = 26; II = bST postpartum, n = 25; III = bST prepartum, n = 27; IV = bST prepartum and postpartum, n = 25). During the prepartum period, cows in groups I and II were not supplemented but those in groups III and IV were supplemented every 2-wk beginning 21 d before expected calving date through calving. During the first 63 DIM only cows in groups II and IV were supplemented with bST. From 64 DIM through the end of lactation cows in all groups were supplemented with the full lactation dose of bST (500 mg/14 d). The BW and BCS were recorded weekly throughout the prepartum and postpartum periods and every 2-wk beyond 70 DIM. Blood samples were collected 3-times a week for analyses of ST and IGF-I. Milk yields were recorded daily though 150 DIM. Prepartum supplementation of bST did not affect BW or BCS, but mean concentrations of ST were increased 12.2% and were 15.5% greater at calving. Overall, mean concentration of IGF-I was not affected by treatment but concentrations were greater at 1 and 2 wk before calving in bST-supplemented cows. During the first 63 DIM the BW and BCS were not affected by treatment. Significant effects of bST-supplementation were detected on concentrations of ST, IGF-I and on milk yield compared to non-supplemented cows in group I. Postpartum concentrations of ST were greater in bST-supplemented cows (TRT II and IV; +41.9 and 54.6%). However, concentrations of IGF-I were greater only in cows in group IV (+25.9%) during the postpartum period. Overall, the three bST-supplemented groups had greater actual milk yield than the control group (I) during the first 63 and 150 DIM. The actual milk yields during 63 and 150 DIM were 6.5 and 4.6 kg/d greater for cows in group IV than cows in group I and the 305-d ME milk yield also was 15.6% greater. No adverse effects of TRT were observed on calf birth weight, colostrum immunoglobulins, ease of calving or other measures evaluated.

Studies on the Hemodilution Perfusion with Rygg-Kyvsgaard Oxygenator (혈액희석 체외순환법에 관한 임상적 관찰 -상온하 Rygg-Kyvsgaard 산화기 및 Sigmamotor pump 사용예를 중심으로-)

  • 손광현
    • Journal of Chest Surgery
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    • v.3 no.2
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    • pp.73-90
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    • 1970
  • Clinical perfusion data on 16 cases of cardiopulmonary bypass using Sigmamotor pump and RyggKyvsgaard Oxygenator which performed at Seoul National University Hospital during the period of Aug. 1968 to Aug. 1970 was analized. AIl cases were hemodiluted and the perfusion was carried out under the normothermic condition. The age of the patients ranged between 6 and 43 years. The b:dy weight varied between 18.3 and 54.0 kg and the body surface area between 0.78 and 1. 59$M^2$. The priming solution was consiste:I with fresh ACD blood. Hartmann solution and Mannitol. The average amount of priming was approximately 2242 ml. The average hemodilution rate was 17%. The flow rate ranged from 1.7L to 3.5L/Min/$M^2$ and averaged 2.4L/Min/$M^2$ or 78mI/Min/kg. The duration of perfusion varied from 22 to 110 min with average of 56.9 minutes. Some hemodynamic responses were observed. The arterial pressure dropped immediately after the initiation of partial perfusion and was more marked after the total perfusion foIlowed by gradual increase to the safety level. The central venous pressure reflected the reduced blood volume especially in the cases of prolonged perfusion which lasted over 60 min. In most of the cases, red blood cell count decreased and white blood ceIl count increased after the perfusion. Hemoglobin level was decreased, averaging of 12.5mg%, Hct 3.3% and platelets count of 18% postoperatively. Plasma hemoglobin increased mildly, from pre-perfusion average value of 4. 06mg% to postperfusion value of 22.5mg%. Serum potassium was 4.4mEq/L pre-operatively and was decreased to 3.7mEq/L postoperatively. Five cases showed definite hypopotassemia immediately after the operation. Sodium and chloride decreased mildly. These electrolyte changes are thought to be related with hemodilution. diuretics and reduced blood volume during and after the perfusion. Arterial blood pH value revealed minimal to moderate elevation from preperfusion average value of 7.376 to 7.461 during perfusion and then 7.365 after perfusion. The pC02 and hicarbonate showed minimal to moderately lowered values. The total CO2 was decreased. Buffer base decreased during perfusion (Av. 42.6mEq/L) and further decreased after the perfusion (Av. 40.8mEq/L). These arterial blood acid base changes suggested that the metabolic acidosis was accompanied by respiratory alkalosis during and immediately after the perfusion. Authors belived that the acidosis could more effectively be corrected with the more additional dose of bicarbonate than we used by this study. The chest tune drainage during the first 24 hours following operation was 1158 ml in average. One case (Case No. 15) showd definite bleeding tendency and it was believed that the cause might be due to the defect of heparin and protamine titration. The average urinary out put during 24 hours post-perfusion was 1291ml. One case (Case No. ]) showed definite post perfusion oliguria. As conclusion hemodilution using fresh ACD blood. Hartmann and Mannitol solution added with Bivon and high flow rate unler normothermia. was thought to amelioratc the severity of mctabolic acidosis during and after perfusion with relatively satisfactory effect on the diuresis and bleeding tendency.

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Effects of insulin and IGF on growth and functional differentiation in primary cultured rabbit kidney proximal tubule cells - Effects of IGF-I on Na+ uptake - (초대배양된 토끼 신장 근위세뇨관세포의 성장과 기능분화에 대한 insulin과 IGF의 효과 - Na+ uptake에 대한 IGF-I의 효과 -)

  • Han, Ho-jae;Park, Kwon-moo;Lee, Jang-hern;Yang, IL-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.783-794
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    • 1996
  • It has been suggested that ion transport systems are intimately involved in mediating the effects of growth regulatory factors on the growth of a number of different types of animal cells in vivo. The functional importance of the apical membrane $Na^+/H^+$ antiporter in the renal proximal tubule is evidenced by estimates that this transporter mediates the reabsorption of approximately one third of the filtered load of sodium and the bulk of the secretion of hydrogen ions. This study was designed to investigate the pathway utilized by IGF-I in regulating sodium transport in primary cultured renal proximal tubule cells. Results were as follows : 1. $Na^+$ was observed to accumulate in the primary cells as a function of time. Raising the concentration of extracellular NaCl induced an decrease in $Na^+$ uptake compared with control cells in a dose dependent manner. The rate of $Na^+$ uptake into the primary cells was about two times higher in the absence of NaCl($40.11{\pm}1.76pmole\;Na^+/mg\;protein/min$) than in the presence of 140mM NaCl($17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$) at the 30 minute uptake. 2. $Na^+$ uptake was inhibited by IAA($1{\times}10^{-4}M$) or valinomycin($5{\times}10^{-6}M$) treatment($50.51{\pm}4.04$ and $57.65{\pm}2.27$ of that of control, respectively). $Na^+$ uptake by the primary proximal tubule cells was significantly increased by ouabain($5{\times}10^{-5}M$) treatment($140.23{\pm}3.37%$ of that of control). When actinomycin D($1{\times}10^{-7}M$) or cycloheximide($4{\times}10^{-5}M$) was applied, $Na^+$ uptake was decreased to $90.21{\pm}2.39%$ or $89.64{\pm}3.69%$ of control in IGF-I($1{\times}10^{-5}M$) treated cells, respectively. 3. Extracellular cAMP decreased $Na^+$ uptake in a dose-dependent manner($10^{-8}-10^{-4}M$). IBMX($5{\times}10^{-5}M$) also inhibited $Na^+$ uptake. Treatment of cells with pertussis toxin(50pg/ml) or cholera toxin($1{\mu}g/ml$) inhibited $Na^+$ uptake. Extracellular PMA decreased $Na^+$ uptake in a dose-dependent manner(1-100ng/ml). 100 ng/ml PMA concentration significantly inhibited $Na^+$ uptake in IGF-I treated cells. However, staurosporine($1{\times}10^{-7}M$) had no effect on $Na^+$ uptake. When PMA and staurosporine were added together, the inhibition of $Na^+$ uptake was not observed. In conclusion, sodium uptake in primary cultured rabbit renal proximal tubule cells was dependent on membrane potentials and intracellular energy levels. IGF-I stimulates sodium uptake through mechanisms that involve some degree of de novo protein and/or RNA synthesis, and cAMP and/or PKC pathway mediating the action mechanisms of IGF-I.

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Nutritional Components and Their Antioxidative Protection of Neuronal Cells of Litchi (Litchi chinensis Sonn.) Fruit Pericarp (리치 과피의 영양화학 성분 및 항산화성 신경세포 보호효과)

  • Jeong, Hee-Rok;Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Kim, Yeon-Su;Jeong, Chang-Ho;Kim, Dae-Ok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.481-487
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    • 2010
  • The nutritional components, antioxidant, and neuroprotective effects of water and a 50% methanol extract from litchi fruit pericarp were investigated. The most abundant mineral, amino acid, and fatty acid were K, proline, and palmitic acid, respectively. In addition, the total water phenolics and 50% methanol extracts were 8.02 and 12.28 mg/g, respectively. The DPPH, ABTS radical scavenging activities and ferric reducing antioxidant power of the water and 50% methanol extracts showed dose-dependent antioxidant activity. In a cell viability assay using MTT, almost all extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase leakage was also inhibited by the pericarp extracts. In particular, the 50% methanol extract showed a higher cell membrane protective effect than the water extract at the highest concentration. Consequently, these data suggest that litchi fruit pericarp can be utilized as an effective and safe functional food substances for natural antioxidants and may reduce the risk of neurodegenerative disorders.

Effects of Palm Kernel Expellers on Growth Performance, Nutrient Digestibility, and Blood Profiles of Weaned Pigs

  • Seo, J.;Kim, W.;Kim, J.;Kim, J.K.;Kim, S.C.;Jang, Y.;Jang, K.;Kim, K.;Kim, B.;Park, S.;Park, I.;Kim, M.K.;Seo, K.S.;Kim, H.B.;Kim, I.H.;Seo, S.;Song, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.7
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    • pp.987-992
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    • 2015
  • This experiment was conducted to investigate the effects of palm kernel expellers on growth performance, nutrient digestibility, and blood profiles of weaned pigs. A total of 88 weaned pigs ($6.94{\pm}0.76kg$ body weight [BW]; 28 d old) were randomly allotted to 2 dietary treatments (4 pigs/pen; 11 replicates/treatment) in a randomized complete block design (sex as a block). The dietary treatments were a typical nursery diet based on corn and soybean meal (CON) and CON added with 20% of palm kernel expellers (PKE). Pigs were fed for 6 wk using a 3-phase feeding program with declining diet complexity and with phases of 1, 2, and 3 wk, respectively. Blood was collected from randomly selected 2 pigs in each pen before weaning and on d 7 after weaning. Pigs were fed respective dietary treatments containing 0.2% chromic oxide from d 29 to 35 after weaning. Fecal samples were collected from randomly selected 2 pigs in each pen daily for the last 3 days after the 4-d adjustment period. Measurements were growth performances, digestibility of dry matter, nitrogen and energy, white and red blood cell counts, packed cell volume, and incidence of diarrhea. The PKE increased average daily gain (ADG) (246 vs 215 g/d; p = 0.06) and average daily feed intake (ADFI) (470 vs 343 g/d; p<0.05) and decreased gain-to-feed ratio (G:F) (0.522 vs 0.628 g/g; p<0.05) during phase 2 compared with CON, but did not affect growth performance during phase 1 and 3. During overall experimental period, PKE increased ADG (383 vs 362 g/d; p = 0.05) and ADFI (549 vs 496 g/d; p<0.05) compared with CON, but did not affect G:F. However, no differences were found on digestibility of dry matter, nitrogen, and energy between CON and PKE. The PKE reduced frequency of diarrhea (15% vs 25%; p = 0.08) for the first 2 wk after weaning compared with CON. Similarly, PKE decreased white blood cells (8.19 vs $9.56{\times}10^3/{\mu}L$; p = 0.07), red blood cells (2.92 vs $3.25{\times}10^6/{\mu}L$; p = 0.09), and packed cell volume (11.1% vs 12.6%; p = 0.06) on d 7 after weaning compared with CON. In conclusion, addition of 20% palm kernel expellers to nursery diet based on corn and soybean meal had no negative effects on growth performance, nutrient digestibility, and blood profiles of weaned pigs.