• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.390-393
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• 2001

The optimal temperature, pH and aeration rate for spore production by Bacillus polyfermenticus SCD in 500 ml shake flask and 5-1 jar fermenter were found to be $32^{\circ}C$, 7.0 and 1.0 vvm. respectively. When batch culture processes was performed under optimized culture conditions. viable cells were $3.3{\times}10^{10}$ CFU/ml and spore cells were $3.3{\times}10^{10}$ CFU/ml. Fed-batch culture processes were also examined with regard to higer maximum viable cell and spore production. The highe viable cells and spores were obtained in 5-1 jar fermenter at 72 h cultivation time by strategy in an intermediate feeding mode with 60% glucose solution 150 ml and 5% soybean flour solution 150 ml fed to the fermenter twice, and the productivity of spore cells was significantly increased. Finally. volumetric productivity of spore cells on fed-batch culture indicated $9.9{\times}10^8$ CFU/ml/h, which was approximately 2 times higher than batch culture. Thus, fed-batch culture show a promise as an industrial production method.

• Food Science of Animal Resources
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• v.40 no.1
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• pp.118-131
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• 2020

It is necessary to add protective agents for protecting the probiotic viability in the preparation process of probiotics starter. In this study, we used whey protein concentrate (WPC), pullulan, trehalose, and sodium glutamate as the protective agent and optimized the proportion of protective agent and spray-drying parameters to achieve the best protective effect on Lactobacillus plantarum. Moreover, the viable counts of L. plantarum in starter stored at different temperatures (-20℃, 4℃, and 25℃) for 360 days were determined. According to response surface method (RSM), the optimal proportion of protective agent was 24.6 g/L WPC, 18.8 g/L pullulan, 16.7 g/L trehalose and 39.3 g/L sodium glutamate. The optimum spray-drying parameters were the ratio of bacteria to protective agents 3:1 (v: v), the feed flow rate 240 mL/h, and the inlet air temperature 115℃ through orthogonal test. Based on the above results, the viable counts of L. plantarum was 12.22±0.27 Log CFU/g and the survival rate arrived at 85.12%. The viable counts of L. plantarum stored at -20℃ was more than 10¹⁰ CFU/g after 200 days.

• International Journal of Oral Biology
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• v.35 no.1
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• pp.1-5
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• 2010

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.52-55
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• 2017

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.430-434
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• 2003

In vivo antagonistic effect of Lactobacillus helveticus CU 631 and Lactobacillus spp. against typical enteritis causing pathogen Salmonella enteritidis KU 101 have been determined, which showed an increase in survival rate and the decline in viable cell numbers of pathogen in liver and spleen at sacrifice. A signifcant difference in the antagonistic effect against KU 101 were observed, which was species and/or strain dependent of Lactobacillus (p<0.01), the survival rate of the mice in the Salmonella infection by feeding L. helveticus CU 631 has been shown to be 157%, whereas those of L. rhamnosus GG ATCC 53103, L. acidophilus ATCC 4356, L. johnsonii C-4 were 137%, 132%, 119% respectively on the basis of lactobacilli non-associated control KU101 fed mice to be 100%. Viable cells of S. enteritidis KU101 in the liver and in the spleen at sacrifice were decreased in Lactobacillus spp. fed group with no significant difference. The higher level of total secretory IgA concentration in the intestinal fluid of lactobacilli fed mice than control mice have been observed. In vitro antagonistic activity of Lactobacillus spp. against KU101 have been determined, a prominent antagonistic activity of CU 631 against KU 101 were demonstrated.

• Proceedings of the Korean Institute of Resources Recycling Conference
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• 2001.05b
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• pp.121-125
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• 2001

For the probiotic feed production, aerobic liquid fermentation of pulverized food wastes was attempted with a yeast Kluyveromyces marxianus. After grinding finely, optimal fermentation conditions of the substrate was investigated by shaking culture. The most active growth of the yeast was shown at solid content of 10%. The proper addition of urea(0.5g/l), o-phosphate(0.4g/l), molasses(4g/l), and yeast extract (1g/1) increased cell growth rate and viable cell count. For optimizing, the nutrients were all added to substrate and fermentation was carried in 2 litre jar fermenter. For the stimulation of hydrolyzing enzyme excretion, mixed culture with Aspersillus oryzae was also conducted. In 12 hours of fermentation, viable cell count of the yeast Kluyveromyces marxianus amounted to the number of 1.4 $\times$10$^{10}$ /1 in the culture medium.

• The Korean Journal of Food And Nutrition
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• v.1 no.2
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• pp.43-49
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• 1988

Fermented barley food was produced by the combining action of an enzyme and a lactobacillus. When Lactobacillus sp. L-5 and commercial liquefying amylase from Tae Pyeong Yang Chemical Co. were selected, inoculated on steamed barley and cultivated at 37$^{\circ}C$ for 48hrs, the fermented product of good quality was obtained. In batch cultivation using rotary drum fermentor, viable cell count reached 1.1$\times$10CFU/g after 12hrs' cultivation, and specific growth rate in logarithmic phase was 0.6hr-1. Viable cell count, acidity, pH, concentration of reducing sugar and viscosity of the 48hrs' fermentation product from rotary drum fermentor was 4.3$\times$108CFU/g, 1.17%, 3.1, 10.7% and 1430cp.

• Preventive Nutrition and Food Science
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• v.10 no.4
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• pp.333-339
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• 2005

Three lactobacillus strains, two from infant feces, and one from cow's milk, were selected among 172 isolates, from multiple sources, for further study based on the antimicrobial activities against six strains of pathogenic bacteria and identified as Lactobacillus acidophilus. The strains revealed a wide scope of spectrum against pathogenic bacteria. Viable Lactobacillus acidophilus KH-l cell counts at pH 2.0 were slightly decreased to $1.42\times10^7$ CFU/mL from $4.18\times10^7$ CFU/mL, while remaining at $3.42\times10^7$ CFU/mL at pH 4.0 with the survival rate of $33.97\%\;and\;81.82\%$, respectively. At the concentration of $0.1\%$ oxgall, L acidophilus KH-l kept growing up to $3.12\times10^7$ CFU/mL with a mean growth rate constant (k) of 0.25, and cell number was slightly decreased to $1.21\times10^7$ CFU/mL (k=0.19) with $0.3\%$ oxgall, but remained at $7.6\times10^6$ CFU/mL (k=0.17) with $0.5\%$ oxgall. L. acidophilus KH-l had a $D_{60}$ value of 7.14, with viable cell numbers $1.4\times10^5$ CFU/mL after heat treatment at $60^{\circ}C$ for 30 minutes. Stability of L acidophilus KH-l at $-20^{\circ}C$ was significantly higher, when the strain was cultivated under the optimum growth temperature $(54.41\%\;and\;54.35\%)$ than at the temperature $(13.53\%)$.

• Korean Journal of Poultry Science
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• v.23 no.1
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• pp.9-17
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• 1996

This study was conducted to compare the survival rate of chick embryos among different eggshell window positions and to search for the most appropriate injection position. The eggshells were punctured at blunt-end, sharp-end and side-up with a sterilized fine forceps, respectively. The survival rate of sharp-end window was higher than the other window positions. Injection of Dulbecco’s modified eagle’s medium (DMEM) through blunt-end window (BE1) was impossible because inner cell membrane was obscure. The 2 ${\mu}$L DMEM was injected into 2.5 d-old embryo blood vessel through sharp end window. To prevent hemorrhages at the point of injection, the air bubbles were injected into the embryo blood vessel. The survival rate of chicks embryo in sharp end window was about 17.0％. Therefore, this sharp-end window system will be helpful for the production of germline chimera or transgenic chicken using primordial germ cells ( PGCs ).

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.