• Journal of Embryo Transfer
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• v.20 no.2
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• pp.169-176
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• 2005

Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.437-442
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• 1996

Two strains of Lactobacillus(L.) casfi and one strain of L. Pentosus, which were isolated from pickles, were used to investigate in studing their characteristics of ${\beta}-galactosidase$. The preferable carbon sources and pH of the MRS media for enzyme production from L. casei No.10 was found to be 1.0% lactose and pH 7.5, from L. Pentosus No.63 was 1.0% galactose and pH 7.5, and from L. casei No.36 was 1.0% lactose and pH 6.5, respectively. The maximum enzyme production from each strain was found after 48 hours culture at $30^{\circ}C$ in a medium with preferable carbon source. The optimum reaction temperature with substrate for ${\beta}-galactosidase$ activity was found at $60^{\circ}C$ for all three strains . The stability of enzyme from L. casei No.36 was found to be at $45^{\circ}C$, from L. Pentosus No.63 was found at $55^{\circ}C$. This stability from L. casei No.36 was found at $40^{\circ}C$, but it was reduced to 60% at $55^{\circ}C$. These stabilities of enzymes remained about 90% at $40^{\circ}C$ for all three strains. The optimal pH for enzyme activities was found to be pH 6.5 for all three strains. Enzyme activity remained over 90% for L. casei No.10 at $pH\;5.0{\sim}6.0$, for L. casei No.36 at $pH\;5.0{\sim}8.0$, and for L. pentosus No.63 at $pH\;6.0{\sim}7.0$.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.317-323
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• 2014

Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.4
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• pp.408-416
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• 2002

A 3D hydrodynamic-ecological coupled model was applied to estimate carrying capacity in Geoje-Hansan Bay where is one of the most important oyster culturing grounds in Korea. We considered the carrying capacity as the difference between food supply to the oysters and food demand, considering monthly difference of the actual growth. The food supply to the system was determined from the results of the model simulation (tidal exchange and chlorophyll $\alpha$) over the culturing period from September to May of the following year. The food demand was estimated from the food concentration (chlorophyll $\alpha$) multiple the filtration rate of oysters that is considered monthly different growth rate of oysters and food concentration. The values of carrying capacity for the system varied from 6.1 ton/ha (minimum carrying capacity) in february to 14.91 ton/ha (maximum carrying capacity) in April of marketable size oysters (>4 g wet-tissue weight) depending on temporal variations in the food supply. The oyster production calculated from present facilities was 9 ton/ha in wet-tissue weight in Geoje-Hansan Bay. This value corresponded to $60\%$ of maximum carrying capacity of the system. The optimal carrying capacity without negatively affecting on oyster production was 5.5 ton/ha when calculated from annual statistic data and 6.1 ton/ha when determined by this study. These results suggest that it must be reduced $32\%$~$39\%$ of oyster facilities in the system.

• Tuberculosis and Respiratory Diseases
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• v.62 no.2
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• pp.98-104
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• 2007

Background: Although there is an increasing incidence of Mycobacterium abscessus pulmonary disease in Korea, the optimal therapeutic regimen has not yet been established and there are no reports of the long-term treatment outcomes. This study examined the long-term treatment outcomes of M. abscessus pulmonary disease. Methods: Twenty-nine patients diagnosed with M. abscessus pulmonary according to the American Thoracic Society criteria and treated from January 1996 to December 2003 were enrolled in ghis study. The clinical characteristics, radiological findings, treatment outcome, and follow up data were analyzed retrospectively. Results: The mean age of the 29 patients was 56.1 (${\pm}13.6$) years and there was a female (22/29) dominance. The chest radiography revealed the nodular bronchiectatic type to be dominant (69%, 20/29). Twenty-seven (93.1%) were prescribed clarithromycin-containing regimens, and injectable drugs, mainly aminoglycosides, were included in the regimen of nineteen patients. The most predominant regimen (48.3%) consisted of clarithromycin and amikacin. The treatment success, failure, and default were achieved in 19(65.5%), 9(31.0%), and 1(3.4%), respectively. The median duration to culture conversion was 42 days (range 15-362) and the median duration of treatment in the success group was 543 days (range 176-1,160). An adjunctive surgical resection was performed in five patients, which resulted in treatment success in two patients. After the completion of treatment, nineteen patients were followed up for a median duration of 931 days (range 230-2,294). Only one (5.3%) patient relapsed 45 days after completing treatment. Conclusion: Treatment with clarithromycin-containing regimens resulted in a successful treatment in approximately two thirds of patients with M. abscessus pulmonary disease. The long-term relapse rate was also quite low.

• Journal of Bio-Environment Control
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• v.27 no.3
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• pp.253-259
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• 2018

This study set out to select a system to realize an optimal environment for strawberry cultivation greenhouses based on data about the growth and development of strawberry and its environment and to provide basic data for the research of its improved productivity. For these purposes, the investigator conducted a field survey with greenhouses for strawberry cultivation in western Gyeongnam. The findings show that farmers in their fifties and sixties accounted for the biggest part in the age groups of strawberry farmers. While those who were under 50 were accounted for approximately 67.5%, those who were 60 or older accounted for 32.5%. As for cultivation experiences, the majority of the farmers had ten years of cultivation experiences or less with some having 30 years of cultivation experiences or more. All the farmers built an arch type single span greenhouse. Those who used nutrient solutions were about 75.0%, being more than those who used soil. All of the farmers that used a nutrient solution adopted an elevated hydroponic system. The single span greenhouses were in the range of 7.5~8.5m, 1.3~1.8m and 2.5~3.5m for width, eaves, and ridge height, respectively, regardless of survey areas. The rafters interval was about 0.7~0.8m. In elevated hydroponic cultivation, the width, height, and interval of the beds were about 0.25m, 1.2m and 1.0m, respectively. As for the strawberry varieties, the domestic ones accounted for approximately 97.5% with Seolhyang being the most favorite one at about 65.0%. As for the internal environment factors of greenhouses, 38 farmers measured only temperature and relatively humidity. As for hydroponics, the farmers used a hydroponics control system. Except for the farmers that introduced a smart farm system for temperature and humidity control, approximately 85.0% controlled temperature and humidity only with a control panel for side windows and ventilation fans. As for heating and heat insulation, all of the farmers were using water curtains with many farmers using an oil or electric boiler, radiating lamp or non-woven fabric, as well, when necessary.

• Journal of Aquaculture
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• v.10 no.4
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• pp.449-456
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• 1997

This study was concuted to determine the optimal conditions for raising the freshwater rotifer, Brachinus calyciflorus. The authors presented some biological informatin obtained from incubation experiment under the various controlled temperatures. Lorica size of the rotifer was divided into two groups : the length and the width for the S-type was $141.0\pm16.7\mu m$($110.1-182.5\;\mu m, n=44$)and $107.0\pm20.3\mu m\;(75.3-152.3\mu m, n=44)$, and those for the L-type was $262.8\pm15.2\mu m\;(234.4-288.6\mu m,\;n=20)\;and\;182.6\pm13.4\mu m (159.8-207.0\mu m,\;n=20$), respectively. The number of eggs being attached on the female varied from 1 to 11 at various culture conditions. Egg type was divided into two groups, large and small. Large and small egg was measured in its major axis as 85a.7-107.8$\mu$m and 55.1-65.2$\mu$m for S-type, and 104.9-121.8 $\mu$m and 62.8-89.1$\mu$m for L-type respectively. The maximum density was reached at 4th day after incubation. The density was 583.9 rotifers/$m\ell$ for $25^{\circ}C$-experimental. group and 421.3 rotifers/$m\ell$ for $22^{\circ}C$-experimental. group respectively. In the case of $28^{\circ}C$-experimental. group, it suddenly decreased into 4.7 rotifers/$m\ell$ at 1st day after incubations and did not recover to its initial density. The maximum rate of increase of populatin per day was reached 0.802 for $22^{\circ}C$-experimental. group at day 2 and fluctuated thereafter. For $25^{\circ}C$-experimental. group it increased to 0.964 at day 3 of incubation and then declined. And the egg ratio of female was reached the maximum of 0.614 for 22$^{\circ}C$- at 3rd day and 0.772 for $25^{\circ}C$-experimental. group at 4th day of incubation.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1133-1140
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• 2006

Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

• Journal of Life Science
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• v.19 no.7
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• pp.994-1002
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• 2009

A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.