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Protoplast Formation and Regeneration of Bacillus strains producing biopolymer (Biopolymer 생산성 Bacillus속 균주의 원형질체 형성과 재생)

  • Yim, Moo-Hyun;Kim, Seong-Ho
    • Applied Biological Chemistry
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    • v.42 no.1
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    • pp.20-28
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    • 1999
  • To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.

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The Effect of some Meteorological Factors on the seed characteristics in Korean White pine (Pinus koraiensis S. et Z.) - The weight of cone and seed per cone - (잣나무 종자형질에 미치는 몇 개 기상인자의 영향 -구과 무게 및 구과당 종자 무게 -)

  • Joo Young-Tuk;Chon Sang-Kuen;Chung Dong-Jun
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.1 no.1
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    • pp.20-28
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    • 1999
  • This study was conducted to reveal the effect of some meterological factors on the weight of cone and seed per cone in Korean white pine (Pinus koraiensis SIEB. et ZUCC.). The weight of cone and seed per cone for 7 years from 1992 to 1998 on 45(1992) year-old trees and some meterological factors for 9 years from 1990 to 1998 were surveyed in Hongcheon-Gun region, Gangweon-Do. Simple correlations and multiple regression between weight of cone and seed per cone and some meterological factors were analyzed. The results obtained from the above experiments were as follows: 1. Positive correlations were found between weight of cone and monthly mean temperature of February in flower bud differentiation year. number of annual hoarfrost days of the cone production year monthly mean temperature of may in the cone production year, as July respectively. There were negative correlations between weight of cone and monthly mean temperature of august in the flowering year, wind speed of April in the flower bud differentiation year, number of clear days of december in the flowering year, number of annual cloudy days of the flowering year, number of precipitation days of june in the flowering year, number of annual precipitation days of the flowering year, number of annual cloudy days of March in the cone production year. number of annual cloudy days from march to October in the flowering year as well as number of precipitation from march to october in the flowering year. 2. Positive correlation between weight of seed per cone and number of hours with sunshine duration of June in the flowering year, the percentage of sunshine duration of June in the flowering year, number of clear days of June in the flowering year. monthly mean temperature of may in the cone production year. as well as monthly mean temperature of July in the cone production year were found. Negative correlations were recognized between weight of seed per cone and monthly mean temperature of January in the flowering year, monthly mean temperature of august in the flowering year, wind speed of april in the flower bud differentiation year, number of annual cloudy days of the flowering year, number of precipitation days of June in the flowering year, number of annual cloudy days from March to October in the flowering year as well as number of precipitation from march to October in the flowering year.

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The evaluation of lens absorbed dose according to the Optimold for whole brain radiation therapy (전뇌 방사선치료 시 Optimold에 따른 수정체의 흡수선량 평가)

  • Yang, Yong Mo;Park, Byoung Suk;Ahn, Jong Ho;Song, Ki Won
    • The Journal of Korean Society for Radiation Therapy
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    • v.26 no.1
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    • pp.77-81
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    • 2014
  • Purpose : In the current whole brain Radiation Therapy, Optimold was used to immobilize the head. However, skin dose was increased about 22% due to the scattering radiation by the Optimold. Since the minimum dose causing cataracts was 2 Gy, it could be seen that the effects were large especially on the lens. Therefore, in the whole brain Radiation Therapy, it was to compare and to evaluate the lens absorbed dose according to the presence of Optimold in the eyeball part. Materials and Methods : In order to compare and to evaluate the lens absorbed dose according to the presence of Optimold in the eyeball part, the Optimold mask was made ??up to 5mm bolus on the part of the eye lens in the human model phantom (Anderson Rando Phantom, USA). In the practice treatment, to measure the lens dose, the simulation therapy was processed by placing the GafChromic EBT3 film under bolus, and after the treatment plan was set up through the treatment planning system (Pinnacle, PHILIPS, USA), the treatments were measured repeatedly three times in the same way. After removing the Optimold mask in the eyeball part, it was measured in the same way as above. After scanning the film and measuring the dose by using the Digital Flatbed Scanner (Expression 10000XL, EPSON, USA), the doses were compared and evaluated according to the presence of Optimold mask in the eyeball part. Results : When there was the Optimold mask in the eyeball part, it was measured at $10.2cGy{\pm}1.5$ in the simulation therapy, and at $24.8cGy{\pm}2.7$ in the treatment, and when the Optimold mask was removed in the eye part, it was measured at $12.9cGy{\pm}2.2$ in the simulation therapy, and at $17.6cGy{\pm}1.5$ in the treatment. Conclusion : In case of removing the Optimold mask in the eyeball part, the dose was increased approximately 3 cGy in the simulation therapy and was reduced approximately 7 cGy in the treatment in comparison to the case that the Optimold mask was not removed. During the whole treatment, since the lens absorbed dose was reduced about 27%, the chance to cause cataracts and side effects was considered to be reduced due to decrease of the absorbed dose to the eye lens which had the high sensitivity on the radiation.

The Effect of Seasoning on the Intestinal Absorption -Absorption by Passive Transport and the Effect of Red Pepper- (조미료가 창자 운동과 흡수기능에 미치는 영향 -소장의 피동적 흡수에 대한 고추의 영향-)

  • Shin, Dong-Hoon;Kim, Joong-Soo;Koh, Jae-Pyong;Ahn, Seung-Woon
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.23-31
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    • 1973
  • Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

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Acid Tolerance of the Acid-Resistant Mutant of Leuconostoc paramesenteroides Improved for Kimchi Starter. (김치 Starter용으로 개량된 Leuconostoc paramesenteroides의 내산성 변이주가 갖는 내산성 특성)

  • 김영찬;정은영;김은해;정대현;최태부;권태종;강상모
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.275-282
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    • 1998
  • To investigate the increased acid tolerance of a acid-resistant mutant Leuconostoc paramesenteroides P-100 improved as a kimchi starter, proton permeability, ATPase acitivity, glycolysis activity, $Mg^2$sup +/ releasement, and membrane fatty acid composition were studied and comprised with its wild type Leuconostoc paramesenteroides Pw. In the proton permeability experiment, the maximum values of the average half time (t$\_$1/2/) of pH equilibration through the cell membrane of the Pw and the P-100 were about 6.4 min and 7.8 min in 150 mM KCI solution, respectively. In the 3% NaCl solution, the t$\_$1/2/ values of the Pw and the P-100 were 5.5 min and 6.9 min, respectively. The values and pHs of maximal specific activities of ATPase originated from the Pw and the P-100 were 0.5 unit/mg protein and 0.78 unit/mg protein at pH 6.0, respectively. The result of pH dependence of glycolysis showed that the P-100 had higher activities than that of Pw except at pH 7.0. The releases of magnesium from the Pw and the P-100 were observed about 54.5% and 23.2% at pH 4.0 after 2 hours, respectively. The results of comparison of membrane fatty acid composition of the Pw with the P-100 showed that C$\_$8:0/, C$\_$9:0/, C$\_$10:0/, C$\_$11:0/, C$\_$18:0/, and C$\_$19:0,cyclo/ were major different fatty acids between two strains and the content of C$\_$18:1/, and C$\_$19:0,cyclo/ were 2.8%, N.D (not detected) in the Pw and 0.4%, 2.3% in the P-100. These results indicated that acid tolerance of the P-100 was significantly improved in comparison with its wild type Pw.

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Operation Parameters on Biological Advanced Treatment of Phenolic High-Strength Wastewater (페놀계 고농도 유기성 폐수의 생물학적 고도처리 운전인자)

  • Hong, Sung-Dong;Park, Chul-Hwi
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.4
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    • pp.797-806
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    • 2000
  • The objectives were to compare the biodegradable threshold concentrations of phenol with the different composition of the influent carbon source and examine the SMA (Specific Methanogenic Activity)and the possibility of simultaneous removal of high-strength organics and nitrogen compounds in UASB(Upflow Anaerobic Sludge Blanket) - PBR(Packed Bed Reactor) process. The results showed that UASB reactors were efficient to remove phenol and phenol + glucose from synthetic wastewater. At phenol conc, of 600 mg/L and SCOD conc. of 2100 mg/L in UASB reactor(with only phenol as substrate), the removal efficiencies of phenol and SCOD were over 99% and 93% respectively, under MLVSS of 20 g. The activity of microorganism was $0.112g\;phenol/g\;VSS{\cdot}d$, $0.351g\;SCOD/g\;VSS{\cdot}d$. The gas production rate was $0.115L/g\;VSS{\cdot}d$ and $CH_4$ content in gas was about 70%. At phenol conc. of 760 mg/L and SCOD conc. of 4300 mg/L in UASB reactor( with phenol + glucose as substrates), the removal efficiencies of phenol and of SCOD were over 99% and 90% respectively, under MLVSS of 20 g. The activity of microoganism was $0.135g\;phenol/g\;VSS{\cdot}d$, $0.696g\;SCOD/g\;VSS{\cdot}d$. The gas production rate was $0.257L/g\;VSS{\cdot}d$ and $CH_4$ content in gas was about 70%. Serum bottle test showed that the activity of granule was inhibited over 1600 mg/L phenol conc, and denitrification and methanogenesis simultaneously took place in UASB granules under co-substrates conditions. PBR reactor packed with cilium type media, was efficient in nitrification. In condition of $0.038kg\;NH_4-N/m^3-media{\cdot}d$. 10~12 mg/L phenol conc. and 200~500 mg/L SCOD conc., nitrification efficiency was over 90% and phenol removal efficiency was over 98%.

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A Study on BMI, Diet Perception and Dietary Behavior of Female University Students (여대생의 체질량지수, 다이어트 인식과 다이어트 행동에 관한 연구)

  • Lee, JinHee
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.3
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    • pp.463-472
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    • 2020
  • The purpose of this study is to identify the BMI, diet perception and diet behavior of female university students. A descriptive survey design was used 165 students from October 29 to November 9, 2018. Data were analyzed using mean, t-test, ANOVA, cross analysis with SPSS 21.0. Total diet perception was 3.44 points, highest sub category was 'practice' with 4.00 points, followed by 'information and education' with 3.58 points, 'necessity' with 3.32 points and lowest category was 'self-recognition' with 2.84. 65 students(43.0%) were 'less than three times' in 'Number of diets' and diet methods was 65(43.0%) for 'food control' and 62(41.1%) for 'exercise'. During in a week, 98 students(84.9%) exercised within three hours, followed by 33(21.9%) who did not exercised at all. 22 students(14.6%) said they experienced damage from dieting, while the types included gastrointestinal disease, menstrual irregularity and hair loss. The total score of diet perception according to BMI was lower in the underweight group than in the normal and overweight groups(p<.001). In diet 'practice' category, low weight group scored lower than normal weight and overweight groups(p<.05). In 'information and education' category, overweight group scored the highest, followed by normal and underweight groups(p<.001). In diet 'necessity' and 'self-recognition' categories, low weight group scored lower than normal weight and overweight groups(p<.001). Diet behavior by diet perception was different in number and method of diet(p<.001). Based on the results, it is thought that attention and education are needed to form a proper diet perception and maintain healthy living for female university students.

Carotenoid Pigments in Echiurid, Urechis unicinctus (개불의 carotenoid 색소성분)

  • KIM Soo Young;HA Bong Seuk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.223-231
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    • 1999
  • Carotenoid pigments of echiurid, Urechis unicinctus were investigated during March, April and May as a part of comparative biochemical studies of carotenoid pigments for the marine organisms other than pisces. Total carotenoid contents were found to be 1.19 mg/100 g in March, 0.98 mg/100 g in April and 0.84 mg/100 g in May, indicating that total carotenoid content was negatively affected by the temperature of sea water that echiurid resided. The carotenoid isolated in March composed of $16.3\%$ diatoxanthin monoester, $14.8\%$ $\beta$-carotene and $12.6\%$ cynthiaxanthin monoester, $8.4\%$ cynthiaxanthin diester, $8.2\%$ zeaxanthin monoester, $7.3\%$ diatoxanthin diester $4.2\%$ astaxanthin $2.9\%$ diatoxanthin, $2.4\%$ triol, $2.3\%$ cynthiaxanthin, $1.7\%$ isocrytoxanthin, $1.5\%$ zeaxanthin diester, $0.8\%$ zeaxanthin and $0.5\%$ lutein. The carotenoid isolated in April composed of $21.9\%$ diatoxanthin monoester, $17.2\%$ cynthiaxanthin monoester and $16.6\%$ $\beta$-carotene $10.9\%$ zeaxanthin monoester, $5.6\%$ cynthiaxanthin diester, $4.9\%$ diatoxanthin diester, $3.1\%$ astaxanthin, $2.4\%$ triol, $2.3\%$ diatoxanthin, $1.7\%$ isocrytoxanthin, $1.5\%$ lutein, $1.1\%$ zeaxanthin, $1.0\%$ cynthiaxanthin and $1.0\%$ zeaxanthin diester. Similarly, the carotenoid isolated in May composed of $25.3\%$ diatoxanthin monoester, $19.7\%$ cynthiaxanthin monoester, $13.0\%$ $\beta$-carotene, and $12.6\%$ zeaxanthin monoester, $5.8\%$ cynthiaxanthin diester, $5.1\%$ diatoxanthin, $3.0\%$ astaxanthin, $2.4\%$ triol, $2.2\%$ diatoxanthin, $1.3\%$ isocrytoxanthin, $1.2\%$ zeaxanthin, $1.1\%$ zeaxanthin diester, $1.0\%$ lutein and $0.9\%$ cynthiaxanthin. Based on these data, monoester-type carotenoids ($37.1\~57.6\%$) and diester-type carotenoids ($11.5\~17.2\%$) of total carotenoids in echiurid were the major carotenoids. Meanwhile, when the sea water temperature was elevated and the contents of total carotenoid in echiurids were decreased, the contents of zeaxanthin monoester, diatoxanthin monoester and cynthiaxanthin monoester were increased, but the contents of zeaxanthin diester, diatoxanthin diester and cynthiaxanthin diseter were decreased, indicating that changes in ester-type caroteoids were differently affected by the sea water temperature.

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Quality Improvement of Rainbow Trout with Pigments and Enzymatic Hydrolysates of Ascidian (Halocynthia roretzi) Tunic 1. Chemical Specificity of Ascidian Tunic and Its Hydrolysates (우렁쉥이 껍질의 색소 및 효소 가수분해물을 이용한 무지개 송어의 품질 향상 1. 우렁쉥이 껍질 및 효소 가수분해물의 화학적 특성)

  • CHOI Byeong-Dae;KANG Seok-Joong;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.3
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    • pp.345-356
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    • 1996
  • Properties of enzymatic hydrolysates from ascidian tunic were assessed on supernatant ratio, solid yields and solid concentration. The concentartion of solid and yields in the extracts were increased as the enzyme concentration raised from $100\;{\mu}l\;to\;1000{\mu}l$ during the extraction period. The optima concentration and reaction time of each enzyme on digestion were $400\;{\mu}l$ 60 minutes, through treated with Duncan's multiple test. The percent of yields of solid, protein and carotenoids for 60 minutes extraction at $400\;{\mu}l$ were $32.32\%,\;1.34\%\;and\;74.60\;mg\%$, respectively, in Viscozyme systems. The extracts were composed with many kinds of carbohydrates such as arabinose, ribose, xylose, galactose, glucose, N-acetyl-D-galactosamine, and N-acetyl-D-glucosamine. Aspartic and glutamic arid were noted as predominant amino acids in all parts. Amino acid profiles of various ascidian tunic part were similiar to each other, but most of essential amino acids content of inter coat was higher than that of root and tunic (body). About sixty six fatty acids components were observed, and their distribution among neutral and polar lipids was compared. The main fatty acids were found to be 14:0, 16:0, 16:1n7, 18:0, 18:1n9, 18:1n7, 18:2n6, 20:5n3, and 22:6n3.

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STUDY ON THE ANCHOVY BOAT SEINE 1. On the Hydrodynamic Resistance and Performance of the Conventional Gear (기선기현강의 연구 1, 재래식 어구의 유구저황과 그물꼴에 관하여)

  • LEE Byoung-gee;YANG Yong-rim;SU Young-tae;SON Boo-il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.4 no.3_4
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    • pp.79-91
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    • 1971
  • A boat seine has been used as a major fishing gear for catching anchovy (Engraulis japonica) in the southern coastal waters of Korea since 1920s. The original seine was operated by two rowing boats as the haul seine. The rowing boats were, in recent, replaced by powered boats. The net size was enlarged by more than three times the original nit as they began to by operated in the deeper waters of approximately sixty meters. However, there are many problems in the efficiency of the fishing gear to be improved. The authors studied on the hydrodynamic resistance and performance of the boat seine net of the 1/10 scale model in tow. The results are summaried as follows. 1. The hydrodynamic resistance converted from model experiment into the full scale is: $$R_1=30,000\;v^{1.2}\;(0.2{\leqq}v{\leqq}1.0)$$ $$R_2=16,000\;v^2\;(0.2{\leqq}v{\leqq}0.6)$$ where $R_1$ and $R_2$ denote the resistance of whole gear and of bag net in kg respectively, and v the speed of flow in m/sec. 2. In the extension wing, approximately seventy percent of the length of the ground rope from the towing end to the inside-wing slopes down from sea level toward the sea bottom, while the thirty percent of the inside of it remains parallel with th: 5:a level. The performance is regarded to b: inefficient for driving fish shoal into the inside-wing, especially for the shoal diving suddenly. 3. At the towing speed higher than 0.2 m/sec, the trailing edge of the inside-wing is blown backward beyond the seaming line connecting the inside-wing and the mouth of the bag net. It is regarded as an unreasonable performance to drive the fish shoal smoothly into the bag net. 4. At the towing speed higher than 0.2 m/sec, the posterior end of the lower bosom is lifted up above the level of the ground rope of the inside-wing. It is considered that the fish shoal diving suddenly can escape through the discrepancy between the lower bosom and the sea bottom, even if the ground rope of the inside-wing sweeps the sea bottom. 5. The angle of inclination of the upper bosom is estimated as $35\~40^{\circ}$. It seems that the inclination is too steep to drive smoothly the fish shoal diving toward the sea bottom into the bag net. 6. In structure, circumference of the posterior section of the bag net is wider by 1.3 times that of the anterior section. Actually in towing at a speed higher than 0.2m/sec, however, the circumference of the posterior section becomes smaller than that of the anterior section. It is recommended to be designed in a long cylindrical form.

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