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Optimization of an Industrial Medium and Culture Conditions for Probiotic Weissella cibaria JW15 Biomass Using the Plackett-Burman Design and Response Surface Methodology

  • Yu, Hyung-Seok;Lee, Na-Kyoung;Kim, Won-Ju;Lee, Do-Un;Kim, Jong-Ha;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.630-637
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    • 2022
  • The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K2HPO4, potassium citrate, ⳑ-cysteine phosphate, MgSO4, and MnSO4) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K2HPO4, 5.0 g/l sodium acetate, 0.1 g/l MgSO4·7H2O, 0.05 g/l MnSO4·H2O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

The effects of synbiotics-glyconutrients on growth performance, nutrient digestibility, gas emission, meat quality, and fatty acid profile of finishing pigs

  • Olivier Munezero;Sungbo Cho;In Ho Kim
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.310-325
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    • 2024
  • Glyconutrients help in the body's cell communication. Glyconutrients and synbiotics are promising options for improving immune function. Therefore, we hypothesized that combining synbiotics and glyconutrients will enhance pig nutrient utilization. 150 pigs (Landrace × Yorkshire × Duroc), initially weighing 58.85 ± 3.30 kg of live body weight (BW) were utilized to determine the effects of synbiotics-glyconutrients (SGN) on the pigs' performance, feed efficiency, gas emission, pork traits, and composition of fatty acids. The pigs were matched by BW and sex and chosen at random to 1 of 3 diet treatments: control = Basal diet; TRT1 = Basal diet + SGN 0.15%; TRT2 = Basal diet + SGN 0.30%%. The trials were conducted in two phases (weeks 1-5 and weeks 5-10). The average daily gain was increased in pigs fed a basal diet with SGN (p = 0.036) in weeks 5-10. However, the apparent total tract digestibility of dry matter, nitrogen, and gross energy did not differ among the treatments (p > 0.05). Dietary treatments had no effect on NH3, H2S, methyl mercaptans, acetic acids, and CO2 emissions (p > 0.05). Improvement in drip loss on day 7 (p = 0.053) and tendency in the cooking loss were observed (p = 0.070) in a group fed basal diets and SGN at 0.30% inclusion level. The group supplemented with 0.30% of SGN had higher levels of palmitoleic acid (C16:1), margaric acid (C17:0), omega-3 fatty acid, omega-6 fatty acid, and ω-6: ω-3 ratio (p = 0.034, 0.020, 0.025, 0.007, and 0.003, respectively) in the fat of finishing pigs. Furthermore, group supplemented with 0.30% of SGN improved margaric acid (C17:0), linoleic acid (C18:2n6c), arachidic acid (C20:0), omega 6 fatty acid, omega-6 to omega-3 ratio, unsaturated fatty acid, and monounsaturated fatty acid (p = 0.037, 0.05, 0.0142, 0.036, 0.033, 0.020, and 0.045, respectively) in the lean tissues of finishing pigs compared to pigs fed with the control diets. In conclusion, the combination of probiotics, prebiotics, and glyconutrients led to higher average daily gain, improved the quality of pork, and more favorable fatty acid composition. Therefore, these results contributed to a better understanding of the potential of SGN combinations as a feed additive for pigs.

Effects of Rare Earth Supplementation on Growth Performance, Blood Immune-Related Cell Population, Meat Quality and Fecal Odor Emission Gases in Finishing Pigs (사료내 희토 첨가가 비육돈의 생산성, 혈액내 면역관련 세포수, 육질특성 및 분 중 악취방출 가스함량에 미치는 영향)

  • Shin, Seung-Oh;Yoo, Jong0Sang;Lee, Je0Hyun;Jang, Hae0Dong;Kim, Hyo0Jin;Huang, Yan;Chen, Ying-Jie;Cho, Jin-Ho;Kim, In-Ho
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.485-498
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    • 2008
  • This study was conducted to evaluate effects of dietary rare earth on growth performance, blood immune- related cell population, meat quality and fecal odor emitting gases in finishing pigs. The total of sixty four (Landrace×Yorkshire×Duroc) pigs(65.42±1.16kg in average initial body weight) were used for feeding trial during 10 weeks of experimental period. Dietary treatments included 1) NC(antibiotic free diet), 2) PC (NC diet+6 weeks 44ppm of tylosin/ 4 weeks 22ppm of tylosin) 3) RE1 (NC diet + 100ppm of RE), 4) RE2 (NC diet+200ppm of RE). There were four dietary treatments with four replicate pens per treatment and four pigs per pen. During the overall periods, there were no significant differences in ADG(Average daily gain), ADFI (Average daily feed intake) and gain/feed ratio among treatments(P>0.05). Dry matter and nitrogen digestibility were higher in RE2 treatment group than other groups(P<0.05). Also, energy digestibility was higher in RE2 treatment group than PC and RE1 treatment groups(P<0.05). At the 6th week WBC(white blood cell) was significantly increased(P<0.05) in RE1 treatment group than NC and RE2 treatment groups. L* value of M. logissimus dorsi muscle color was significantly increased(P<0.05) in rare earth supplemented groups compared to NC treatment group(P<0.05). However, a* value was lower in RE1 treatment group than PC treatment group (P<0.05). In fatty acid composition of Intramuscular fat, total MUFA was significantly higher in RE2 treatment group than other groups(P<0.05). Also, total UFA was significantly increased in RE2 treatment group compared with NC and PC treatment groups(P<0.05). In fatty acid composition of back fats, total SFA of rare earth supplemented groups were lower than in PC treatment group(P<0.05). UFA:SFA ratio was significantly higher in rare earth supplemented groups than PC treatment group(P<0.05). In fecal odor emission, NH3 was significantly decreased(P<0.05) in rare earth supplemented groups compared to NC and PC treatment groups. In conclusion, the results of the experiment was affected by rare earth supplementation on digestibilities, meat quality, fatty acid and fecal odor emission gases in finishing pigs.

Effects of Growth and Cellular Tissue under Abnormal Climate Condition in Chinese Cabbage (이상기상 조건이 배추의 생육 및 세포조직에 미치는 영향)

  • Lee, Sang Gyu;Choi, Chang Sun;Choi, Jun Myung;Lee, Hee Ju;Park, Suhyoung;Do, Kyung Ran
    • Journal of Bio-Environment Control
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    • v.22 no.2
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    • pp.87-90
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    • 2013
  • The average annual and winter ambient air temperatures in Korea have risen by $0.7^{\circ}C$ and $1.4^{\circ}C$, respectively, during the last 30 years. Due to climate change, the occurrence of abnormal weather conditions has become more frequent, causing damage to vegetable crops grown in Korea. Hot pepper, chinese cabbage and radish, the three most popular vegetables in Korea, are produced more in the field than in the greenhouse. It has been a trend that the time for field transplanting of seedlings is getting earlier and earlier as the spring temperatures keep rising. Seedlings transplanted too early in the spring take a longer time to resume the normal growth, because they are exposed to suboptimal temperature conditions. This experiment was carried out to figure out the change of cellular tissue of chinese cabbage under the condition of low temperature to provide the information regarding the coming climatic change, on the performance of 'Chunkwang' chinese cabbage during the spring growing season. In our study, plant height, number of leaf, chlorophyll and leaf area was lower at the open field cultivation than heating house treatment after transplanting 50 days. Especially in fresh weight, compared with heating treatment, open field and not heated treatment were notably low with the 1/3 level. Of damage symptoms due to low temperature cabbage leaves about 10 sheets when $-3.0^{\circ}C$ conditions in chinese cabbage was a little bit of water soaking symptoms on the leaves. $-7.4^{\circ}C$ under increasingly severe water soaking symptoms of leaf turns yellow was dry. Microscopy results showed symptoms of $-3.0^{\circ}C$ when the mesophyll cell of palisade tissue and spongy tissue collapse, $-7.4^{\circ}C$ palisade tissue and spongy tissue was completely collapsed. The result of this study suggests that the growers should be cautioned not to transplant their chinese cabbage seedlings too early into the field, and should be re-transplanting or transplanting other plants if chinese cabbage are exposed to suboptimal temperature conditions ($-3.0^{\circ}C$ or $-7.4^{\circ}C$).

Growth of Radish Plug Seedlings as Influenced by Various Pre-planting Nitrogen Levels in Inert Media (기비로 혼합된 질소 수준이 무 플러그 묘 생장에 미치는 영향)

  • Sung, Jwa Kyung;Park, Myong Sun;Choi, Jong Myung
    • Journal of Bio-Environment Control
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    • v.26 no.3
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    • pp.201-207
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    • 2017
  • Objective of this research was to investigate the influence of pre-plant nitrogen levels in root media on plug seedling growth of radish cv. Soksungbommu. To achieve the research purpose, a root medium, the mixture of perlite, coir dust, and peatmoss (volume percentage of 30:35:35) was formulated and the N levels incorporated during mixing were controlled to 0, 100, 250, 500, 750, 1,000, and $1,500mg{\cdot}L^{-1}$. Then, the seeds were sown into 72-cell plug trays in which the root medium was packed. The measurements of growth and analysis of tissue and root media were conducted 2 and 4 weeks after sowing. Elevation of pre-plant N levels raised EC and turned down pH of root media. But, as seedling grew, the pH rose and EC get lowered in all treatments. The EC as well as $NH_4-N$ and $NO_3-N$ concentrations of root media declined gradually until week 2, but those declined sharply between weeks 2 to 4. The seedling growth 2 weeks after sowing showed quadratic response to pre-plant N levels with the highest growth in $250mg{\cdot}L^{-1}$ treatment and lagging growth in the treatments of lower or higher N levels than $250mg{\cdot}L^{-1}$. The seedling growth 4 weeks after sowing showed also quadratic response with the highest growth in $500mg{\cdot}L^{-1}$ treatment. The tissue N contents were get higher and those of K, Ca, and Mg were get lower as pre-plant N levels were elevated. Above results suggest that lower than $250mg{\cdot}L^{-1}$ in pre-plant N levels is optimistic for growth of plug seedling and avoidance of toxic injury in very young stage.

Light Quality and Photoperiod Affect Growth of Sowthistle (Ixeris dentata Nakai) in a Closed-type Plant Production System (밀폐형 식물생산시스템에서 광질과 광주기에 따른 씀바귀의 생육)

  • Kim, Hye Min;Kang, Jeong Hwa;Jeong, Byoung Ryong;Hwang, Seung Jae
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.67-76
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    • 2016
  • This study was conducted to examine the optimal environmental condition for promoting the growth of sowthistle as affected by light quality and photoperiod in a closed-type plant production system. Seeds were sown in 240-cell plug trays and then germinated for 3 days at a 24-hour photoperiod in a closed-type plant production system with LED lights (R:B:W = 8:1:1). Seedlings were transplanted and grown under 3 types of LED (R:B:W = 8:1:1, R:W = 3:7, or R:B = 8:2) and 4 photoperiods (24/0, 16/8, 8/16, or 4/20 hours) with $230{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light intensity at a density of $20cm{\times}20 cm$. The experimental design was a randomized complete block design. Plants were cultured for 40 days un der the condition of $21{\pm}2^{\circ}C$ and $70{\pm}10%$ relative humidity after transplanting. Plants were fed with a recycling nutrient solution (pH 7.0 and EC $2.0dS{\cdot}m^{-1}$) contained in a deep floating tank. Fresh weight and dry weight of shoot or root, leaf length, and leaf area were the greatest in the photoperiod of 24/0 (light/dark) with RW LED. The highest number of leaves occurred in the photoperiod of 16/8 (light/dark) with RB LED, while the incidence of tip burn was higher in the photoperiod of 24/0 (light/dark) compared to the other treatments. Chlorophyll value was the highest in the 16/8 (light/dark) photoperiod and there was no significant difference by light quality. Chlorophyll fluorescence was the lowest in the photoperiod of 24/0 (light/dark) compared with other treatments. Therefore, in terms of economic feasibility and productivity for Ixeris dentata Nakai cultivation in a closed-type plant production system, the results obtained suggest that plants grew the best when kept in a photoperiod of 16/8 (light/dark) and light quality of combined LED RW (3:7).

Do Paneth Cells Regulate the Zinc Body Burden? (Zinc 대사와 관련된 Paneth 세포활성의 변화에 관한 조직화학적 연구)

  • Jo, Seung-Mook;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.357-365
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    • 2000
  • Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.

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Effect of Aster scaber and Ixeris dentata on Contractility and Vasodilation of Cardiovascula and Endothelial Cell in Hyperlipidemic Rat (참취 및 씀바귀 첨가식이가 고지혈증 흰쥐의 심혈관 수축과 이완 및 혈관내피세포에 미치는 영향)

  • Lim, Sang-Sun;Lee, Jong-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.300-307
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    • 1997
  • The effects of Aster scaber and Ixeris dentata on cadiovascular system in hyperlipidemic rats were examined. Five groups of thirty Sprague Dawley rats were fed with the diet contained 1% cholesterol, 0.25% sodium cholate, 10% coconut oil and 5% lard(control group) for 4 weeks. Each experimental diet group was added with 5% plant powder or extract of the 5% plant powder by dry weight. Contractile or relaxation responses in the isolated artria and thoracic aortae were measured and the morphological changes of the aortic endotherium from the rats fed the experimental diet were inspected. In response to isoproterenol, the number of right atrial spontaneous beat was significantly lower in Cham chyi powder group$(PP_{1})$ and Sumbagui powder group$(PP_{2})$ than control at $10^(-8)M$ concentration. The contraction forces by injection of phenylephrine and calcium in isolated thoracic aorta were significantly low in each experimental groups compared with the control. The relaxation rates by acetylcholine represented comparatively higher value in $PP_{1}$ than control. The morphological changes of endothelial cell surface was a little in $PP_{1}$ and $PP_{2}$ compared with control, while the damages were considerably advanced in Cham chyi and Sumgbagui extract diet group$(PE_{1},\;PE_{2})$.

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Cultural Characteristics of Lactobacillus amylovorus IMC-1 Producing Antibacterial Substance (항균성 물질을 생산하는 Lactobacillus amylovorus IMC-1의 배양학적 특성)

  • Mok, Jong-Soo;Song, Ki-Cheol;Kim, Young-Mog;Chang, Dong-Suck
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.249-254
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    • 2002
  • To determine the abilities as both lactic starter and probiotics for fermented foods, we investigated the potency of acid production, proteolytic activity and lactose metabolism of Lactobacillus amylovorus IMC-1. And the strain was cultured with lactococci in 10% skim milk medium. It was also examined the bactericidal action of antibacterial substance, produced by the strain IMC-1, against pathogenic bacteria. L. amylovorus IMC-1 showed excellent production of acid in 10% skim milk supplemented with yeast extract, and produced 0.8 and 2.7% of acid at 12 and 72 h incubation, respectively. It was found that the activity of ${\beta}-galactosidase$, about $39\;{\mu}M/minute/dry$ cell weight (mg), was stronger than that of $phospho-{\beta}-galactosidase$ in the strain IMC-1. The strain showed weak proteolytic activity in 10% skim milk, thus it produced 6 and $69\;{\mu}g/mL$ of free tyrosine at 12 and 72 h cultivation, respectively. It was known that the strain utilized mainly ${\alpha}-casein$ than ${\beta}-casein$ from patterns of SDS-PAGE. Mixed culture produced more acid than single cultures of L. amylovorus IMC-1 and Streptococcus thermophilus NIAI 510. Single culture of Str. thermophilus and mixed culture showed increasing cheese flavor with incubation times. Optimal fermentation time of mixed culture for the acid production and flora of lactic starter was 16 and 12 h by adding 0.1 and 0.5% of yeast extract to 10% skim milk, respectively. Antibacterial substance produced by the strain IMC-1 reduced about 2 log of the viable cell counts of both Escherichia coli O157 and Shigella flexneri after 24 and 4 h incubation, and they were not detected after 48 and 6 h incubation, respectively.

Enhanced PHB Accumulation in Photosystem- and Respiration-defective Mutants of a Cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis sp. PCC 6803의 에너지 대사 결함 돌연변이 균주에서의 Poly(3-hydroxybutyrate) 축적량 증진)

  • Kim Soo-Youn;Choi Gang Guk;Park Youn Il;Park Young Mok;Yang Young Ki;Rhee Young Ha
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.67-73
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    • 2005
  • Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.