• Food Engineering Progress
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• v.21 no.2
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• pp.143-149
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• 2017

This study was carried out to investigate antimicrobial activity and characteristics of Asparagus cochinchinenesis which was steamed and fermented with lactic acid bacteria. A. cochinchinensis was prepared to steaming process which was washed and freeze dried. A. cochinchinensis was steamed at $95^{\circ}C$ for 12 h and dried by hot air at $50^{\circ}C$ for 24 h. After steaming process, A. cochinchinensis was fermented with lactic acid bacteria (Leuconostoc mesenteroides 4395, Lactobacillus sakei 383 and Lactobacillus plantarum KCCM 11322). Ethyl acetate extracts of fermented A. cochinchinensis had antimicrobial activities for the respiratory disease bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Escherichia coli). A. cochinchinensis had highest antimicrobial activity for the P. aeruginosa which fermented with L. mesenteroides 4395. The minimum inhibition concentration (MIC) of A. cochinchinensis fermented with L. mesenteroides 4395 was 10 mg/mL for S. aureus, S. epidermidis, E. coli and 5 mg/mL for P. aeruginosa. The MIC of A. cochinchinensis fermented with L. sakei 383 and A. cochinchinensis fermented with L. plantarum KCCM 11322 were the same. Total sugar was decreased from $863.33{\pm}17.47mg/mL$ to $722.67{\pm}5.51mg/mL$ during the steaming process. But reducing sugar was increased from $99.36{\pm}1.32mg/mL$ to $109.29{\pm}2.71mg/mL$ during the steaming process. Total sugar was decreased to 301.50-361.42 mg/mL and reducing sugar was decreased to 27.39-62.20 mg/mL during the fermentation process.

• Journal of Korean Society of Forest Science
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• v.109 no.4
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• pp.382-389
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• 2020

Soil properties in opencast mines are a key factor in reclamation (revegetation) of mining areas. In this study we determined the soil physical and chemical properties of kaolinite tailings, reclaimed areas, and adjacent natural red pine (Pinus densiflora S. et Z.) forests in kaolinite opencast mines in Sancheong-gun, Gyeongsangnam-do. Six sites were chosen for collection of soil samples to determine soil physical and chemical properties at a soil depth of 10 cm. Soil bulk density was significantly higher (P < 0.05) in the kaolinite tailings (1.51 g·cm-3) than in the reclaimed areas (1.19 g·cm-3) and red pine forests (0.93 g·cm-3), whereas air phase in the kaolinite tailings (14.2%) was significantly lower than in the red pine forests (32.6%). Clay content in the red pine forests was significantly higher than in the reclaimed areas (18.7%) or kaolinite tailings (14.8%), whereas soil structural stability index was significantly lower in the reclaimed areas (1.61%) and kaolinite tailings (0.87%) than in the red pine forests (7.75%). Soil pH was significantly higher in the kaolinite tailings (pH 6.68) and reclaimed areas (pH 6.27) than in the red pine forests (pH 5.31). Soil organic carbon and total nitrogen were significantly higher in the red pine forests (C: 36.03 mg·g-1; N: 2.08 mg·g-1) than in the reclaimed areas (C: 5.00 mg·g-1; N: 0.31 mg·g-1) than in the kaolinite tailings (C: 2.12 mg·g-1; N: 0.07 mg g-1). The amount of available phosphorus was not significantly different among the three treatments. The concentration of exchangeable potassium was significantly lower in the kaolinite tailings (0.08 cmolc·kg-1) than in the reclaimed areas (0.21 cmolc·kg-1) and red pine forests (0.30 cmolc·kg-1). These results indicate that, because of high soil bulk density and low soil organic carbon, total nitrogen, available phosphorus, and exchangeable potassium in kaolinite tailings and reclaimed mining areas, soil nutrient management is needed in order to reclaim the vegetation in these type of areas.

• Maritime Security
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• v.1 no.1
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• pp.215-240
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• 2020

In 2017, the U.S. DARPA coined 'mosaic warfare' as a new way of warfighting. According to the Timothy Grayson, director of DARPA's Strategic Technologies Office, mosaic warfare is a "system of system" approach to warfghting designed around compatible "tiles" of capabilities, rather than uniquely shaped "puzzle pieces" that must be fitted into a specific slot in a battle plan in order for it to work. Prior to cover mosaic warfare theory and recent development, it deals analyze its background and several premises for better understanding. The U.S. DoD officials might acknowledge the current its forces vulnerability to the China's A2/AD assets. Furthermore, the U.S. seeks to complete military superiority even in other nation's territorial domains including sea and air. Given its rapid combat restoration capability and less manpower casualty, the U.S. would be able to ready to endure war of attrition that requires massive resources. The core concept of mosaic warfare is a "decision centric warfare". To embody this idea, it create adaptability for U.S. forces and complexity or uncertainty for the enemy through the rapid composition and recomposition of a more disag g reg ated U.S. military force using human command and machine control. This allows providing more options to friendly forces and collapse adversary's OODA loop eventually. Adaptable kill web, composable force packages, A.I., and context-centric C3 architecture are crucial elements to implement and carry out mosaic warfare. Recently, CSBA showed an compelling assessment of mosaic warfare simulation. In this wargame, there was a significant differences between traditional and mosaic teams. Mosaic team was able to mount more simultaneous actions, creating additional complexity to adversaries and overwhelming their decision-making with less friendly force's human casualty. It increase the speed of the U.S. force's decision-making, enabling commanders to better employ tempo. Consequently, this article finds out and suggests implications for Korea armed forces. First of all, it needs to examine and develop 'mosaic warfare' in terms of our security circumstance. In response to future warfare, reviewing overall force structure and architecture is required which is able to compose force element regardless domain. In regards to insufficient defense resources and budget, "choice" and "concentration" are also essential. It needs to have eyes on the neighboring countries' development of future war concept carefully.

• Journal of Bio-Environment Control
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• v.32 no.2
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• pp.165-171
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• 2023

Potato dry rot is one of the potato storage diseases caused by Fusarium species and is a representative pathological disorder that induced post-harvest loss during storage. Chlorpropham treatment for sprouting inhibition is mainly used for room temperature storage of potatoes for processing. In this study, the inhibitory effect of chlorpropham on Fusarium-induced dry rot of potato 'Dano'. To investigate the mycelial growth rate of the dry rot fungus (Fusarium solani Appel & Wollenw), mycelial growth was investigated in a chlorpropham (5.0, 50.4, 503.8, and 5,038 ppm) and prochloraz (0.1, 1.0, 10.0, and 100.0 ppm) medium containing F. oxysporum mycelia. Mycelia were more inhibited as the concentration of chlorpropham and prochloraz increased during incubation at 20℃, and the inhibition rate was 98.2% and 100% when treated with 503.8 ppm of chlorpropham and 10ppm of prochloraz in 14 days, respectively. Potato Dano tubers inoculated with F. oxysporum were dipped in chlorpropham (5.0, 50.4, and 503.8 ppm) and prochloraz (100 ppm) to investigate the effect of preventing dry rot during cold storage at 20℃ and 4℃ in vivo. The disease diameter of potatoes stored at room temperature (about 20℃) was reduced to 13.0 mm in the prochloraz 100 ppm teatment, and 10.7 mm in the chlorpropham 50.4 ppm treatment compared to 13.7 mm in the control tuber at 70 days of storage. The disease progression in all treatments including control was similar with no statistically significant difference at 4℃ air temperature. From the results of this study, it is considered that treatment with 50.4 ppm of chlorpropham after harvest will be useful for suppressing dry rot of stored potatoes.

• Korean journal of applied entomology
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• v.13 no.3 s.20
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• pp.105-133
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• 1974

The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

• The Korean Journal of Physiology
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• v.15 no.1
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• pp.27-36
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• 1981

Relatively little has been done on the metabolic changes of the lung produced by the excessive alcohol ingestion to the point of the acute alcohol intoxication. In the present study, an effort was made to clarify the possible changes of the pulmonary surfactant system by the acute alcohol ingestion. The dynamic pulmonary compliance and the levels of protein and inorganic phosphorus (Pi) of both lung lavage and extract were chosen as the parameters of the pulmonary surfactant activities. The albino rats of both sexes were used, and 1.5 ml of 50% ethanol per 100 g body weight was given by oral intubation, and the experiment was performed at 1, 3, 6, 12, and 24 hours after the alcohol ingestion. The rat was sacrificed by cutting the carotid arteries, and blood sample for the determination of hematocrit(Hct) and the blood alcohol concentration was obtained. Both lungs were completely removed without dammage to the lung tissue, and the pulmonary compliance was measured by the changes of pressure-volume(P-V) curves by inflating or deflating the lung with air. Immediately after the P-V curves were recorded, the lung lavage was obtained by washing the lobes with 15ml of isotonic saline 3 times with a syringe. Next, total lungs were homogenized and filtered to obtain the lung extract. The protein and Pi levels were measured using the lung lavage and extract as the samples, and the lung/body weight ratio(L/B ratio) was also calculated. The results thus obtained were compared with the normal values and summarized as follows. The blood alcohol concentration reached the highest level of $0.71{\pm}0.02\;g\;%$ at 1 hr and gradually decreased until 24 hrs$(0.36{\pm}0.02\;g%)$ after the alcohol ingestion, but all the experimental groups showed significant increase comparing with the normal. The highest Hct value was obtained at 1hr$(64.86{\pm}2.45%)$ and significantly elevated value was continued throughout the experiment. The L/B ratio was significantly lowered from 3hrs until 24hrs after the alcohol ingestion but from 6 th hr on, a generally elevated value was observed with a significant value at 12 hrs and gradual recovery to the normal value at 24 hrs after the alcohol ingestion. The pulmonary compliance at inflation and deflation did not change appreciablly from the normal until 3 hrs after the alcohol ingestion but from 6 th hr on, a generally elevated value was observed with a significant value at 12 hrs and gradual recovery to the normal value at 24 hrs after the alcohol ingestion. The protein level of the lung lavage stowed a significantly increased value of $12.36{\pm}0.35\;mg/gm(3rd hr)$, $12.70{\pm}0.74\;mg/gm(12 th hr)$, and $12.65{\pm}0.88\;mg/gm(24 th hr)$, respectively, comparing with the normal value of $10.65{\pm}0.62\;mg/gm$, and the Pi level also showed a similar tendency of significant increase at 12th hr $(7.65{\pm}0.63\;{\mu}mol/gm)$ and 24 th hr$(6.70{\pm}0.36\;{\mu}mol/gm)$ comparing with the normal value of $5.32{\pm}0.20\;{\mu}mol/gm$. The protein level of the lung extract in the alcohol group was generally similar to the normal value with a slight decrease at 1st and 3 rd hr, tut the Pi level of the lung extract was generally increased in the alcohol group, and a significant increase was observed at 6 th hr$(17.77{\pm}1.54\;{\mu}mol/gm)$, 12 th hr$(13.92{\pm}0.78\;{\mu}mol/gm)$ and 24 th hr$(14.57{\pm}0.53\;{\mu}mol/gm)$ of the alcohol ingestion comparing with the normal value of $10.34{\pm}0.37\;{\mu}mol/gm$. From the above, it may be concluded that the acute alcohol intoxication produces the metabolic changes of the lungs by the increased surfactant activities and elevated pulmonary compliance.

• Korean Journal of Soil Science and Fertilizer
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• v.35 no.2
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• pp.93-104
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• 2002

A field survey and experiment was conducted from 1996 to 1998 to develop rational technology for turfgrass vegetation of runway side of Incheon International Airport on the reclaimed tidal land in Young-Jong Island. Backfill of the experimental site was finished on August 1995. The experimental site was 8 ha located in the middle of the construction place for the main parking lot in front of the terminal building construction. The experimental field was drained by main open ditch, and divided three main plots, no subsurface tile drain, subsurface tile drain spacing with 22.5m, and with 45 m, respectively. The 17 sub plots were designed to test the effect of soil covering with red earth loam by 5 cm and 20 cm depth, application of chemical compound fertilizers and livestock manures, dressing of artifical soils and hydrophylic soil conditioners. The tested turfgrasses were three transplanting indigenous turfgrasses, Zoysia koreana, Zoysia sinica and Zoysia japonica, and two hydroseeding mixed exotic turgrasses, cool type I(tall fescue 30%, kentucky blue grass 40%, perenial ryegrass 30%), and cool type II(tall fescue 40%, perenial ryegrass 20%, fine fescue 20%, alkaligrass 20%). The soil backfilled with dredged seasand was sand textured with high salt concentration and low fertility. The soil showed high pH, low organic matter and low available phophate contents. The percolation rate was fast with high hydraulic conductivity. Desalinization was fast after installation of the main open drainage system. No subsurface tile drainage effect was found showing little difference in turfgrass growth. The covering and visual growth of turfgrasses were the best in the 20-cm soil covering with compound fertilizer treatment. The covering and visual growth of turfgrasses were satisfactory in the 5 cm soil covering with compound fertilizer treatment and with livestock manure treatments. The hydrophillic soil conditioner treatments were effective but expensive at present. The coverage and visual quality of turfgrasses were good for Zoysia koreana and Zoysia japonica. The coverages of turfgrasses by the hydroseeding with the mixed exotic turfgrasses were less than transplanting of native turfgrasses. In conclusion, for the runway side vegetation purposes, the subsurface tile drainage might not necessary as main open ditch drainage be sufficient due to fast percolation rate of the backfilled dredged seasand. The 5 cm soil covering with red earth might be sufficient for the runway side, but the 20 cm soil covering might be necessary for the runway side where high density of turfgrass coverage was necessary to protect from the airplance air blow.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.3 no.3
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• pp.177-186
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• 1970

The spawning of the abalone, Haliotis discus hannai, was induced In October 1969 by air ex-position for about 30 minutes. At temperatures of from 14.0 to $18.8^{\circ}C$, the youngest trochophore stage was reached within 22 hours after the egg was laid. The trochophore was transformed into the veliger stage within 34 hours after fertilization. For $7\~9$ days after oviposition the veliger floated in sea water and then settled to the bottom. The peristomal shell was secreted along the outer lip of the aperture of the larval shell, and the first respiratory pore appears at about 110 days after fertilization. The shell attained a length of 0.40 mm in 15 days, 1.39 mm in 49 days, 2.14 mm in 110 days, 5.20 mm in 170 days and 10.00 mm in 228 days respectively. Monthly growth rate of the shell length is expressed by the following equation :$L=0.9981\;e^{0.18659M}$ where L is shell length and M is time in month. The density of floating larvae in the culture tank was about 10 larvae per 100 co. The number of larvae attached to a polyethylene collector ($30\times20\;cm$) ranged from 10 to 600. Mortality of the settled larvae on the polyethylene collector was about $87.0\%$ during 170 days following settlement. The culture of Nauicula sp. was made with rough polyethylene collectors hung at three different depths, namely 5 cm, 45 cm and 85 cm. At each depth the highest cell concentration appeared after $15\~17$ days, and the numbers of cells are shown as follows: $$5\;cm\;34.3\times10^4\;Cells/cm^2$$ $$45\;cm\;27.2\times10^4\;Cells/cm^2$$ $$85\;cm\;26.3\times10^4\;Cells/cm^2$$ At temperatures of from 13.0 to $14.3^{\circ}C$, the distance travelled by the larvae (3.0 mm In shell length) averaged 11.36 mm for a Period of 30 days. Their locomation was relatively active between 6 p.m. and 9 p.m., and $52.2\%$ of them moved during this period. When the larvae (2.0 mm in shell length) were kept in water at $0\;to\;\~1.8^{\circ}C$, they moved 1.15cm between 4 p.m. and 8 p.m. and 0.10 cm between midnight and 8 a.m. The relationships between shell length and body weight of the abalone sampled from three different localities are shown as follows: Dolsan-do $W=0.2479\;L^{2.5721}$ Huksan-do $W=0.1001\;L^{3.1021}$ Pohang $W=0.9632\;L^{2.0611}$

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.1-7
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• 2002

The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.17-23
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• 2002

The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.