• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.6
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• pp.812-813
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• 1995

Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.223-231
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• 1983

The productions of beta-exotoxin from sixteen Bacillus thuringiensis strains were examined by Micrococus flava primarily, and then measured by spectrophotometer during culturing in Conner and Hansen mineral salts medium at 28$^{\circ}C$. Also the toxic effects of the toxin to mice were checked. The growth of Bacillus thuringiensis K2 and BTK2-T1, -T13, -T33 and -T40 got into stationary phase at 6 hour culture and then maintained it up to 48 hours without severe fluctuation. The production of beta-exotoxin from the strains, BTK2, BTK2-T1, -T13, -T17 and -T33 appeared at 6 hour culture and the amounts of the toxin were about 40 $\mu\textrm{g}$/$m\ell$ at 6 hour culture, approximately 70 $\mu\textrm{g}$/$m\ell$ at 12 hours, approximately 85$\mu\textrm{g}$/$m\ell$ from 24 hours to 48 hours. At 48 hour-culture, BTK2 produced 80 $\mu\textrm{g}$/$m\ell$ of beta-exotoxin (5.5$\times$10$^{8}$ cells/$m\ell$, BTK2-T13 produced 84 $\mu\textrm{g}$/$m\ell$ (4.3$\times$10$^{8}$ cells/$m\ell$), BTK2-T17 produced 87$\mu\textrm{g}$/$m\ell$ (1.4$\times$10$^{8}$ cells/$m\ell$), and BTK2-T33 produced 84 $\mu\textrm{g}$/$m\ell$ (4.9$\times$10$^{8}$ cells/$m\ell$). All other serotypes also produced beta-exotoxin. At 48 hour culture, BTK-37 produced 88$\mu\textrm{g}$/$m\ell$ (6.1$\times$10$^{8}$ cells/$m\ell$), BTK-35 produced 81 $\mu\textrm{g}$/$m\ell$), and the rest of them produced less than 70 $\mu\textrm{g}$/$m\ell$. To check the toxicity of beta-exotoxin and B. thuringiensis, the cultured media with microorganisms were inoculated to mice by per os, intraperiloneal, subcutaneous and intracerebral injection, and nasal cavity inoculation for 30 days. However, the toxin did not kill all of the treated mice.

• Korean Journal of Environmental Biology
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• v.18 no.4
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• pp.403-409
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• 2000

Surface and bottom water samples were collected from 10 stations located in the coastal area of Tongyong in April, August and October 2000. Distribution of heterotrophic bacteria, coliform bacteria and fungi in the sea water samples was investigated by measuring the corresponding viable cell number according to the plate counting method. Heterotrophic bacteria in the surface water counted 3.1$\times$10$^2$- 4.0$\times$10$^3$cfu ml$\^$-1/, 2.7$\times$10$^3$- 1.2$\times$10$\^$5/cfu ml$\^$-1/ and 1.3$\times$10$^2$- 7.2$\times$10$^2$cfu ml$\^$-1/ in April, August and October, respectively. The cell number of coliform bacteria in the surface water amounted to 0-1.5$\times$10$^1$cfu ml$\^$-1/, 3.5$\times$10$^1$- 5.2$\times$10$^3$cfu ml$\^$-1/ and 0-1.8$\times$10$^2$cfu ml$\^$-1/ in April, August and October, respectively. As for fungi, the cell number in the surface water was 0-3.0$\times$10$^1$propagules ml$\^$-1/, 3.0$\times$10$^1$- 8.0$\times$10$^1$ propagules ml$\^$-1/ and 0-2.2$\times$10$^1$ propagules ml$\^$-1/ in April, August and October respectively. The surface water samples from the station 3 in August were added with feed stuffs for fish as much as 0.01 gl$\^$-1/, 0.1 gl$\^$-1/ and 1 gl$\^$-1/ and cultured at 5$\^{C}$, 15$\^{C}$, 25$\^{C}$ and 35$\^{C}$. Microbial cells were not isolated at all when the culturing temperature was 5$\^{C}$. However, the microbial cell number increased significantly in all the surface water samples containing 1 gl$\^$-1/ of the feed stuffs when cultured at 15$\^{C}$, 25$\^{C}$ and 35$\^{C}$

• Journal of Embryo Transfer
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• v.23 no.1
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• pp.43-49
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• 2008

This study was carried out to examine the efficiency of biopsy methods, and the pregnancy rate, calving and abortion rates, gestation length and birth weight of Hanwoo calves following transfer of fresh, frozen and sexed Hanwoo embryos produced in vitro. The survivability of biopsied embryos was 80.0 and 90.0% using aspiration and punching methods at 24 h after culturing, respectively. The ratios of male and female embryos were 42.1 and 52.6%, respectively, and the percentage of sex unidentified was 5.3%. Pregnancy rates was not significantly different between hCG and control group (46.4 vs. 38.5%), fresh and frozen embryos (41.3 vs. 35.0%), and sexed and IVP embryos (27.5 vs. 41.2%) (p>0.05). Calving and abortion rates of IVP and sexed embryos were not significantly different in calving (85.0 vs. 87.0%) and in abortion (15.2 vs. 13.3%) (p<0.05). Gestation length of IVP and sexed calves were 281.3 and 288.2 days in female and 283.0 and 282.3 days in male, and the birth weight of IVP and sexed calves were 23.6 and 25.0 kg in female and 24.6 and 23.8 kg in male, respectively. There were no difference in gestation length and birth weight between IVP embryos and sexed embryos (p>0.05). Administration of hCG to recipients did not improve the pregnancy rate following transfer of Hanwoo embryos produced in vitro and sexed embryos. Although the production of calves derived from sexed Hanweoo embryos cultured in vitro can be obtained, the efficiency of sexed calves production need to be improved in biopsy methods and pregnancy rate. Further study should be focused on the improvement of pregnancy rates for commercial application of embryo transfer.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.35 no.2
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• pp.136-146
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• 1999

The telemetry system for the oxygen, pH, turbidity and the distribution ecology of fishes was constructed by the authors in order to product and manage effectively in shallow sea culture and setnets fisheries, and then the experiments for the telemetry system carried out at the culturing fishing ground in coast of Sanyang-Myon, Kyoungsangnam-Do and the set net fishing ground located Nungpo bay in Kojedo province respectively from October, 1997 to June 1998.As those results, the techniques suggested in the telemetry system for which find out the relationship between the physical and chemical environment in the sea and the distribution ecology of fishes gave full display its function, and its system could be operated as real time system. This research can also provide base-line data to develope a hybrid system unifying the marine environment information and the fisheries resources information in order to manage effectively coastal fishing ground.

• Korean journal of food and cookery science
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• v.15 no.6
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• pp.555-564
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• 1999

The objective of this study was to investigate the preference of root vegetables and the inhibitory effect of the vegetables on harmful enzymes of intestinal bacteria. Two hundred fifty respondents in Seoul area surveyed to obtain information from Sep. 30 to Oct. 30, 1998. Respondents preferred Inpuomoea batatas (sweet potato, 4.05), Solanum tuberosum(potato, 3.97), Allium cepa(onion, 3.68), Codonopsis lanceolata(3.64) and Raponus sativus(redish, 3.60). The growth of B. breve K-110 was effectively increased by adding 0.5% extract of Solanum tuberosum(139%), Codonopsis lamceolate(145%), Dioscorea japonica(164%), Colocisia antiquorum(144%) extract to the medium. B. breve K-100 for beneficial bacteria, and E. coli HGU-3 or Bacteroides JY-6 for harmful bacteria were used to determine the inhibitory effect of root vegetables on harmful intestinal enzymes after co-culturing harmful and beneficial bacteria. The extract of Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica (yam) and Colocisia antiquorum (taroes) showed inhibitory effect on ${\beta}$-glucuronidase and tryptophanase of intestinal bacteria. The macromolecules were isolated from Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica and Colocisia antiquorum by Sephadex G-100 column chromatography. By adding these isolated marcromolecules to the medium, the growth of B. breve K-100 were also increased and high inhibitory effects on the ${\beta}$-glucuronidase and tryptophanase were measured. These results suggested that the harmful enzymes of intestinal bacteria were inhibited by consuming Solanum tuberosum, Codonopsis lanceolata, Dioscorea japonica and Colocisia antiquorum. Therefore, they could prevent gastrointestinal diseases.

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.545-553
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• 2019

This study was conducted to determine whether the water in which nitrate accumulated during long-term fish culture in an aquaponics system without water exchange could be removed and reused as catfish-culturing water. The catfish (Silurus asotus) were cultured in the urban aquaculture system using BFT (Biofloc Technology) aquaculture and an aquaponics system (two rearing tanks, 3 tons each) without exchanging the rearing water. After 151 days (from March to August) of rearing, 2.8 g of fry had grown to an average weight of 171.3 g (total weight, 56.53 kg) and 235.5 g (total weight 71.1 kg), respectively. The overall survival rate was 65% in the urban aquaculture system. However, the survival rate was 77.7% before separation into the two tanks. The survival rates after the separation were 92.9% and 78.0%. In the early biofloc watermaking process, there was a high mortality rate. After water stabilization, the mortality rate decreased and some mortality occurred during the period when the total amount of suspended solids (TSS) increased. The results of monthly blood analysis of the catfish showed that the AST concentration was significantly higher in April. Blood ALT levels and triglycerides showed no difference in the rearing period and the glucose, cholesterol, and total protein levels were significantly higher in July. There was no difference in the other periods. The plants produced by the aquaponics system using catfish-rearing water were lettuce, basil, chard, and red chicory. These showed smooth growth and a total of 148.85 kg of plants were harvested in five months. It was possible to remove nitric acid from the aquaponics system and reuse it as catfish-rearing water. Maintaining proper plant quantity according to the capacity of the catfish showed that the combination of agricultural and aquatic products was possible.

• Applied Microscopy
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• v.36 no.3
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• pp.141-153
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• 2006

The purpose of this study is to find out how soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth and research the result of using MPS, suspected to demage eye cells, on rabbit eye's corneal epithelium and endothelium tissue. In this treatise, $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) had been selected among the MPS. After culturing L929 roil line, cell growth inhibition rate was measured by MTT assay, and by making Hematoxylin and Eosin stain specimen. the morphology was observed by optical microscope. In the In vivo experiment,9 white rabbit eyes (18 eyes) were classified into 3 groups. The experimental group is left eyes (9 eyes) of rabbit, and MPS were dropped; however. the control group, the right eyes (9 eyes), were only used a saline solution without preservatives. After the dropping within the period, the cornea surface of rabbit eyes were stained by Rose bengal and observed. To figure out the changes of the corneal epithelium and endothelium tissue scanning electron microscopy (SEM) has been used. As the result, the rate of cell growth inhibition was 54%. 73% and 36%, respectively. Morphological changes represented that the shape has been changed into oval or round shape and those are not considered as a common formation of L929 cell line. When it comes to staining Rose bengal, each experiment group was stained red which is not shown in controls. The polygonal mosaic pattern of a corneal epithelium was disturbed in the picture taken by SEM; furthermore, the shape of the corneal endothelium was irregular. In conclusion, as we consider antimicrobial effect and the safety on living cells, it is necessary that we should improve concentration of preservatives and study continuously to develop a new preservatives without a toxic effect on the cornea surface.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.503-513
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• 1986

Previous studies have developed the technique of topical application of tetracycline(TC) into the periodontal pockets and examined the change of clinical parameters and subgingival microbial morphotypes. The purpose of this study was to longitudinally examine the clinical and microbiological effects of topically applied TC in a double-blind and split-mouth design. Thirteen patients with moderate periodontitis, who were treated with or without TC application and scaling treatment, were examined. TC gel(3%) was used to apply into the selected periodontal pockets twice a week for 2 weeks. During the experiment, clinical parameters and subgingival microbial morphotypes were examined, and for isolation of black-pigmented Bacteroides(BPB) and streptococci, an anaerobic sample culturing was done at week 0, 2, and 7. In clinical observation the TC-scaled group exhibited a significant decrease of Gingival Inflammatory Index, Plaque Index, Sulcus Bleeding Index, pocket depth, and gingival crevicular fluid when compared to the TC-unsealed, placebo-scaled, and placebo-unsealed groups. The result of microbial morphotype observation showed a significant increase of coccal form and a decrease of spirochetes in the TC-scaled, TC-unscaled, and placebo-scaled groups. The culture study of streptococci revealed that TC with scaling treatment resulted in a significant increase of S. sanguis I at week 2, but its proportion had returned to the base line level. The anaerobic culture study showed that BPB was significantly reduced in the TC-scaled and TC-unsealed groups at week 7. Among BPB species, B. intermedius declined significantly with time treatment(week 2 and 7) in the TC-scaled and TC-unsealed groups. These results suggest that the settled pathogenic microflora can be succeeded by nonpathogenic microflora in periodontal pockets after TC treatment.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.169-175
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• 2006

The objective of this study was carried out to examine the polar body extrusion of in vitro matured porcine follicular oocytes as a non-invasive marker of oocyte quality to know the developmental competence in advance. The porcine oocytes matured for 48 hours were examined the polar body extrusion and some parts were stained. The examined oocytes were matured for additional $16{\sim}18$ hours and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 hours for diploid formation. The treated oocytes were washed and cultured for 7 days. The polar body extrusion and degeneration rates were varied with $9.9{\sim}52.4%$ and $21.4{\sim}61.8%$ by repetition. The polar body extruded oocytes were shown the polar body chromosome and metaphase II plate by staining. However the non-extruded oocytes were shown expanded nucleus with 39.1%, premature chromosome condensation with 19.6%, metaphase I plate with 10.9 %, metaphase II with 13%, condensed chromatin with 6.5%, and absent nuclear material with 8.7%. The oocytes that were not examined for the polar body extrusion were cleaved 45.0%, and developed to blastocyst stage with 11.3%. In examined oocytes for polar body extrusion,. the polar body extruded oocytes were cleaved 94.2% and developed with 42.5%. This result suggests that discarding of the degenerating oocytes and oocytes that not extruded polar body will be effective for experiments of culturing effect in porcine embryos and embryo biotechnology.