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Characterization and Purification of Subtilosin A Produced by Bacillus vallismortis MCBL 1012 Isolated from Seasoned Dried Radish

  • Se-Yeon Lee;Dae-Ook Kang
    • Journal of Life Science
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    • v.34 no.8
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    • pp.576-587
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    • 2024
  • In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO4 and NiSO4 at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

Effects of Replacing Soy Protein Concentrate with Fermented Soy Protein in Starter Diet on Growth Performance and Ileal Amino Acid Digestibility in Weaned Pigs

  • Min, B.J.;Cho, J.H.;Chen, Y.J.;Kim, H.J.;Yoo, J.S.;Wang, Q.;Kim, I.H.;Cho, W.T.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.99-106
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    • 2009
  • For Exp. 1, 120 ((Yorkshire${\times}$Landrace)${\times}$Duroc) weaned pigs (7.96${\pm}$0.01 kg average initial BW, 21 days weaning) were used in a 28 d-growth assay to determine the effects of replacing soy protein concentrate (SPC) with fermented soy protein (FSP) in a starter diet (d 0 to 7) on the growth performance, apparent fecal amino acid digestibility and subsequent performance in weaned pigs. Dietary treatments included: i) FSP0 (basal diet; whey-skim milk powder-SPC based diet); ii) FSP5 (replacing SPC with 5% FSP); iii) FSP10 (replacing SPC with 10% FSP). Pigs were fed the phase I diet for 7 days, and then each group was fed a common commercial diet for 21 days to determine the effect of previous diet on subsequent performance. Average daily gain (ADG) from d 5 to 7 (linear effect, p = 0.01) and d 7 to 14 (linear effect, p<0.001) were increased as FSP level increased. The pigs fed with FSP was heavier than the pigs fed with SPC at d 5 to 7 and d 7 to 14 after weaning (p<0.05). In the entire period (d 0 to 28), there were no significant differences in weight gain and final weight between SPC and FSP diets (p>0.05). Average daily feed intake (ADFI) was higher in pigs fed with the 5% FSP diet than those fed with the other diets at d 0 to 2 post-weaning (quadratic effect, p = 0.05). Also, for the entire period of phase I (d 0 to 7), pigs consumed more 5% FSP diet compared to other treatments (quadratic effect, p = 0.03). Gain/feed (G/F) was not affected by dietary SPC or FSP in phase I and subsequent periods, but G/F from d 5 to 7 after weaning was improved linearly (p = 0.04) as dietary FSP level increased. Pigs fed with 10% FSP also improved G/F compared with those fed only SPC (p<0.05). At d 7, there were linear increments in fecal dry matter (DM) (p<0.1) and nitrogen (N) (p<0.01) digestibilities as the dietary FSP level increased. The digestibilities of fecal essential and total amino acids were increased as the FSP level increased (linear effect, p<0.1). For Exp. 2, three ((Yorkshire${\times}$Landrace)${\times}$Duroc) weaned barrows (average initial BW of 7.32 kg) were surgically fitted with a simple T-cannula approximately 15 cm prior to the ileo-cecal junction. The experimental designs were 3${\times}$3 latin squares with pigs and periods as blocking criteria. Dietary treatments and composition were the same as in Exp. 1. Apparent ileal N digestibility was increased as FSP level was increased (linear effect, p<0.05). The dietary treatments (SPC and FSP) did not affect apparent ileal DM digestibility (p>0.05). Among essential amino acids, apparent digestibility of ileal arginine (Arg), lysine (Lys), methionine (Met) and phenylalanine (Phe) were improved as the FSP level increased (linear effect, p<0.1). Also, apparent ileal total essential, non-essential and total amino acid digestibilities were increased linearly (p<0.1). In conclusion, replacing SPC with fermented soy protein appeared beneficial in growth performance, N and amino acid digestibility during the early 7 days after weaning, and an equivalent effect showed on growth performance in subsequent period of 7 to 28 days after weaning.

On the Milled Wood Lignins Isolated from Hardwood by Progressive Milling (단계적(段階的) 분쇄법(粉碎法)에 의해 조제(調製)된 활엽수(闊葉樹) MWL에 관한 연구(硏究))

  • Cho, Nam Seok
    • Journal of Korean Society of Forest Science
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    • v.45 no.1
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    • pp.62-67
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    • 1979
  • Ultraviolet microscopy of ultrathin sections of wood has proved to be one of the useful means for determining the lignin distribution in the various regions of the cell wall. Also, spectral approach and quantitative analysis of isolated compound middle lamella fraction from birch xylem have revealed that the lignin associated with the vessel secondary wall and middle lamella is composed predominantly of gualacylpropane units. Lignin deposited in the fiber and ray parenchyma secondary walls is composed mostly of syringylpropane units. The middle lamella lignin around fibers and ray cells contains both guaiacyl and syringyl propane quits. On the basis of the results above, this research was carried out to clarify the origin of milled wood lignin (MWL) by analysing the chemical characteristics of ML MWLs extracted at various milling stages. The amount of phenolic hydroxyl-, ${\alpha}$-carbonyl-, and methoxyl-group in the MWL's increases the milling time. And progressive mining contributes to the merease of ratio of syringylaldehyde to vanillin(S/V ratio) after nitrobenzene oxidation of MWL. Accordingly, It could be concluded that milled wood lignin extracted at the initial milling stage derives from compound middle lamella region of cell wall, whereas, with progressive milling, lignin of secondary wall of fiber is introduced gradually to milled wood lignin. These results are suggesting that heterogeneous chemical structure of lignins in hardwood exists. Although milled wood lignin at the initial stage seems to have lower molecular weight in comparison with milled wood lignin extracted at final milling stage from the result of Gel-filtration curves, further study would be required on molecular weight distribution of milled wood lignin in future.

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Nutritional Value of Soybean Meal from Various Geographic Origin and Effect of Their Dietary Supplementation on Performance of Broilers (원산지가 다른 대두박의 영양적 가치평가와 육계의 생산성에 미치는 영향)

  • Lee, Bo-Keun;Kim, Jae-Young;Kim, Ji-Sook;You, Sun-Jong;An, Byoung-Ki;Kim, Eun-Jib;Kang, Chang-Won
    • Journal of Animal Science and Technology
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    • v.51 no.3
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    • pp.217-224
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    • 2009
  • This study was carried out to investigate the nutritional value of soybean meal (SBM) from various geographic origins and the effects of their dietary supplementation on performance of broiler chickens. Nutritional value of dehulled SBM originating from USA, and non-dehulled SBM from India (IND), Argentina (ARG) and Korea (KOR) were evaluated by analyzing chemical composition, urease activity (UA) and KOH solubility, and determining true metabolizable energy (TME), nitrogen corrected TME (TMEn) and true amino acid availability (TAAA). The contents of crude protein ranged from 45.43% (ARG) to 48.47% (USA) and those of crude fiber varied widely from 3.48% (USA) to 7.12% (IND). The measurements of lysine varied from 2.79% (IND) to 3.09% (USA) and those of methionine from 0.56% (IND) to 0.65% (USA). The values of TMEn varied from 2986.6 kcal/kg (IND) to 3228.9 kcal/kg (USA) and the averages of TAAA were from 91.61% (IND) to 92.27% (USA). UA was found to be from 0.02 (ARG) to 0.04 (KOR, USA) and those of KOH solubility from 73% (ARG) to 84% (USA). A total of four hundred 20-days-old male broiler chicks were divided into four groups and fed with isocaloric and isonitrogenous experimental diets containing 27.5% of SBM and same amounts of lysine and sulfur amino acids for 15 days. Final body weight and body weight gain were the highest in birds fed with SBM from USA and lowest in birds fed with SBM from IND although the differences were not statistically significant. The feed/gain in chicks fed diet containing SBM from USA was significantly improved (p<0.05) compared to those of the other groups. There were no significant differences in carcass characteristics and the concentration of total cholesterol in serum among the treatments. The results of in vitro assay and bioassay agreed with the performance of broiler chicks, and thus there were close correlation between the broiler performance and the measured nutritive values of SBM. In conclusion, dehulled SBM from USA was superior to non- dehulled SBM from ARG and IND with regard to nutritive values.

Effects of Dietary Wild-Ginseng Adventitious Root Meal on Growth Performance, Blood Characteristics and Meat Quality in Growing-Finishing Pigs (산삼 부정근박 급여가 육성-비육돈의 생산성, 혈액성상 및 육질특성에 미치는 영향)

  • Jang, Hae-Dong;Hahn, Eu-Joo;Jeon, Won-Kyung;Paek, Kee-Yeoup;Kim, Hyo-Jin;Shin, Seung-Oh;Kim, In-Chul;Park, Jun-Chul;Kim, Jin-Dong;Kim, In-Ho
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.677-686
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    • 2008
  • This study was conducted to evaluate the effects of dietary wild-ginseng adventitious root meal on growth performance, blood characteristics and meat quality characteristics in growing-finishing pigs. Ninety six pigs[(Landrace×Yorkshire)×Duroc] with average initial body weight of 68.29±0.31kg were used in 70d growth trial. Dietary treatments included 1) CON(Basal diet), 2) WGR1(Basal diet+0.5% wild- ginseng adventitious root meal), 3) WGR2(Basal diet+1.0% wild-ginseng adventitious root meal) and 4) WGR3(Basal diet+1.5% wild-ginseng adventitious root meal). The pigs were allotted into four dietary treatments with six replicate pens and four pigs per pen in a completely randomized design. For the whole period, final body weight and ADG were increased in CON treatment compared to WGR3 treatment(Linear effect, P=0.005). In blood characteristics, red blood cell(RBC) was significantly increased in CON and WGR2 treatments compared to WGR1 treatment (Quadratic effect, P=0.019). WGR2 treatment resulted in higher white blood cell(WBC) than CON and WGR1 treatments(Linear effect, P=0.041). WBC difference was significantly improved in WGR2 treatment compared to other treatments (Linear effect, P=0.042). Total protein was increased in WGR2 treatment compared to CON treatment (Quadratic effect, P=0.011). In cholesterol concentration of blood, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride were not significantly different among treatments. In meet quality, pH in WGR1 treatment was higher than WGR3 treatment(Quadratic effect=0.022). Water holding capacity(WHC) was significantly increased in WGR2 treatment compared to WGR3 treatment(Quadratic effect, P=0.050).

Effects of Precooling and Packaging Methods on Quality of Asparagus Spears during Simulated Distribution (아스파라거스의 모의 유통 과정에서 예냉 방법과 포장 조건이 품질에 미치는 영향)

  • Yoon, Hyuk Sung;Choi, In-Lee;Han, Su Jung;Kim, Ju Young;Kang, Ho-Min
    • Journal of Bio-Environment Control
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    • v.27 no.1
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    • pp.7-12
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    • 2018
  • This study was conducted to determine effects of precooling and storage methods on asparagus spears' quality such as changes of fresh weight and color during simulated export distribution. Two types of precooling methods, air cooling and hydrocooling, were applied prior to packaging by comparing with no precooling as a control. Asparagus spears were packed with oxygen transmission rate (OTR) film for modified atmosphere packaging (MAP) and perforated (PF) film for a conventional packaging. All treatments were stored at $8^{\circ}C$ for 20 hours, and subsequently at $4^{\circ}C$ by final storage day, which is simulated distribution temperature condition from Yanggu, Korea to Shimonoseki, Japan. The half cooling time was 12 minutes for air cooling and 15 seconds for the hydrocooling, indicating precooling process of asparagus spears faster with the hydrocooling. Rates of respiration and ethylene production were lowest with hydrocooling. Fresh weight loss was higher, approximately 11%, at the control condition in conventional storage, compared with the MAP, less than 0.5%. Carbon dioxide and oxygen content in the MAP was in the permissible ranges for asparagus spears under recommended CA/MA conditions under both the air cooling and hydrocooling. Ethylene content in the film package was lower with the precooling treatment. Firmness of stems was lowest with the hydrocooling prior to the MAP. Visual quality, off-odor, and hue angle value were best with hydrocooling prior to the MAP. In conclusion, the combination of hydrocooling with the MAP is effective in preserving quality during the export distribution process.

Stabilization of Radioactive Molten Salt Waste by Using Silica-Based Inorganic Material (실리카 함유 무기매질에 의한 폐용융염의 안정화)

  • Park, Hwan-Seo;Kim, In-Tae;Kim, Hwan-Young;Kim, Joon-Hyung
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.5 no.3
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    • pp.171-177
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    • 2007
  • This study suggested a new method to stabilize molten salt wastes generated from the pyre-process for the spent fuel treatment. Using conventional sol-gel process, $SiO_2-Al_2O_3-P_2O_5$ (SAP) inorganic material that is reactive to metal chlorides were prepared. In this paper, the reactivity of SAP with the metal chlorides at $650{\sim}850$, the thermal stability of reaction products and their leach-resistance under the PCT-A test method were investigated. Alkali metal chlorides were converted into metal aluminosilicate($LixAlxSi1-_xO_{2-x}$) and metal phosphate($Li_3PO_4\;and\;Cs_2AlP_3O_{10}$) While alkali earth and rare earth chlorides were changed into only metal phosphates ($Sr_5(PO_4)_3Cl\;and\;CePO_4$). The conversion rate was about $96{\sim}99%$ at a salt waste/SAP weight ratio of 0.5 and a weight loss up to $1100^{\circ}C$ measured by thermogravimetric analysis were below 1wt%. The leach rates of Cs and Sr under the PCT-A test condition were about $10^{-2}g/m^2\;day\;and\;10^{-4}g/m^2\;day$. From these results, it could be concluded that SAP can be considered as an effective stabilizer for metal chlorides and the method using SAP will give a chance to reduce the volume of salt wasteform for the final disposal through further researches.

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Effects of Dietary Bacillus subtilis Supplementation on Meat Quality, Growth Performance and Fecal Malodor Gas Emission in Finishing Pigs (Bacillus subtilis 급여가 비육돈의 육질 특성, 생산성 및 분내 유해가스 발생에 미치는 영향)

  • Jang, Hae-Dong;Yoo, Jong-Sang;Lee, Je-Hyun;Kim, Hyo-Jin;Shin, Seung-Oh;Hwang, Yan;Zhou, Tain-Xiang;Chen, Ying-Jie;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.327-332
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    • 2008
  • This study was conducted to evaluate the effects of dietary Bacillus subtilis on meat quality, growth performance and fecal malodor gas emission in finishing pigs. Thirty-six pigs (Landrace ${\times}$ Yorkshire ${\times}$ Duroc, $83.53{\pm}1.01\;kg$ average initial body weight) were used in a 35 d growth assay. Dietary treatments were 1) CON (basal diet), 2) B1 (basal diet + B. subtilis 0.1%) and 3) B2 (basal diet + B. subtilis 0.2%). The pigs were distributed into four pigs per pen with three replicate pens per treatments by completely randomized design. For the entire period, the final weight, ADO, ADFI and gain/feed were not significantly different among the treatments. There were no significant differences in meat quality (sensory evalution, meat color, TBARS, water holding capacity, drip loss, cooking loss and M. longissimus dorsi area) among the treatments. $H_2S$ was significantly decreased in B2 treatment compared to CON and B1 treatments (p<0.05). However, ammonia, mercaptans and acetic acid were not significantly different among the treatments. In conclusion, B. subtilis 0.2% treatments decreased fecal $H_2S$ gas emission in finishing pigs.

Changes in water quality and hematological parameters according to the stocking density of olive flounder, Paralichthys olivaceus raised in bio-floc environment (바이오플락 양식기술 적용 넙치, Paralichthys olivaceus 수용밀도에 따른 수질변화 및 혈액학적 성상 변화)

  • Kim, Jun-Hwan;Jeong, Eun-Ha;Kim, Seok-Ryel;Kim, Su Kyoung;Kim, Su-Kyoung;Hur, Young Baek
    • Korean Journal of Environmental Biology
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    • v.37 no.2
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    • pp.155-163
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    • 2019
  • The Paralichthys olivaceus (mean weight 34.3±3.5 g) was used in the conduction of density experiment in this study to establish the optimal density determination for a period of 13 weeks. The density consisted of four groups. These were group 1 (500 fish, 40 fish m-2), group 2 (750 fish, 60 fish m-2), group 3 (1,000 fish, 80 fish m-2) and group 4 (1,250 fish, 100 fish m-2), each in 12.56 ㎡ circular water tanks. In the high density groups, such as groups 1 and 2, the nitrite was increased to over 21 mg L-1 (group 3) and 25 mg L-1 (group 4). The experiment of the groups was terminated by skin ulcers and mortality after 49 days for group 3 and 43 days for group 4. The water quality environment, such as the water temperature, dissolved oxygen, salinity and pH, remained constant. The ammonia and nitrite in groups 1 and 2 remained stable after ammonia and nitrite peak. However, the experiments in groups 3 and 4 were terminated due to the mortality induced by high nitrite. Hematological parameters, such as hemoglobin and hematocrit, were significantly decreased in the high density groups. The plasma components were significantly changed in the high density groups, such as groups 3 and 4. The mean weight of groups 1 and 2 after 13 weeks were 91.3 g and 83.7 g, respectively. The survival rates were 99.4% and 98.2%, respectively. The final FCR was 0.6 in both groups. The results of this study show that the density of 80 fish m-2 or more induce mortality due to high nitrite with hematological changes. Additionally, they indicate that the 60 fish m-2 indicate proper density in bio-floc environment in olive flounder weighing less than 100 g.

Effects of Dietary Effective Microorganism (EM) on Growth Performance, Microflora Population and Noxious Gas Emission in Broiler (유용 미생물 첨가가 육계 생산성, 맹장 내 균총 및 유해가스 발생량에 미치는 영향)

  • Kang, Hwan Ku;Kim, Chan Ho;Bang, Han Tae;Kim, Ji Hyuk;Kim, Min Ji;Kim, Dong Woon;Na, Jae Cheon;Hwangbo, Jong;Yang, Young Rok;Choi, Hee Cheol;Moon, Hong Kil
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.227-233
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    • 2014
  • This study investigated the effects of dietary supplementation with the effective microorganism (EM) on the growth performance, blood parameter, small intestinal microflora, and noxious gas emission of broilers. A total 720 1-d old ROSS 308 was randomly assigned to 6 dietary treatment groups: control, virginiamycin (6 mg/kg), 0.1% PB 0.1% EM, 0.5% EM, and 1.0% EM. Each treatment was fed to 4 replicates of 30 birds per diet for d 35. Two-phase feeding program with a starter diet from 0 to 3 wk, and a finisher diet from 4 to 5 wk was used in the experiment. Within each phase, a diet was formulated to meet or exceed NRC requirements of broilers for macro- and micronutrients. The diet and water were available ad libitum. Result indicated that during overall periods of the experiment, final weight, body weight gain, and feed intake were not different among dietary treatments. Feed conversion ratio was less (P<0.05) for EM treatments than control, antibiotics, and PB. Total cholesterol (TC), triglyceride (TRG), glucose (GLU), total protein (TP), calcium (CA), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were not different among dietary treatments. White blood cell (WBC), hemoglobin (Hb), heterophils (HE), lymphocyte (LY), monocytes (MO), and eosinophils (EO) were not different among dietary treatments. HE:LY was less (P<0.05) for EM0.5 treatments than control, antibiotics, and PB. Lactobacillus was greater (P<0.05) for EM treatments than control and antibiotics. E. coli and Salmonella were not different among dietary treatments. $NH_3$ and $CO_2$ wereless (P<0.05) for EM treatments than control. These results indicated that EM treatments were effective feed conversion ratio, noxious gas emission and micro flora population on the cecum in broilers.