• The Korean Journal of Pesticide Science
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• v.13 no.1
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• pp.1-12
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• 2009

To assess the effect of butachlor on freshwater aquatic organisms, acute toxicity studies for algae, invertebrate and fishes were conducted. The algae grow inhibition studies were carried out to determine the growth inhibition effects of butachlor (Tech. 93.4%) in Pseudokirchneriella subcapitata (formerly knows as Selenastrum capriconutum), Desmodesmus subspicatus (formerly known as Scendusmus subspicatus), and Chlorella vulgaris during the exposure period of 72 hours. The toxicological responses of P. subcapitata, D. subspicatus, and C. vulgaris to butachlor, expressed in individual $ErC_{50}$ values were 0.002, 0.019, and $10.4mgL^{-1}$, respectively and NOEC values were 0.0008, 0.0016, and $5.34mg\;L^{-1}$, respectively. P. subcapitata was more sensitive than any other algae species. Butachlor has very high toxicity to the algae, such as P. subcapitata and D. subspicatu. In the acute immobilisation test for Daphnia magna, the 24 and $48h-EC_{50}$ values were 2.55 and $1.50mg\;L^{-1}$, respectively. As the results of the acute toxicity test on Cyprinus carpio, Oryzias latipes and Misgurnus anguillicaudatus, the $96h-LC_{50}s$ were 0.62, 0.41 and $0.24mg\;L^{-1}$, respectively. The following ecological risk assessment of butachlor was performed on the basis of the toxicological data of algae, invertebrate and fish and exposure concentrations in rice paddy, drain and river. When a butachlor formulation is applied in rice paddy field according to label recommendation, the measured concentration of butachlor in paddy water was $0.41mg\;L^{-1}$ and the predicted environmental concentration (PEC) of butachlor in drain water was $0.03 mg\;L^{-1}$. Residues of butachlor detected in major rivers between 1997 and 1998 were ranged from $0.0004mg\;L^{-1}$ to $0.0029mg\;L^{-1}$. Toxicity exposure ratios (TERs) of algae in rice paddy, drain and river were 0.004, 0.05 and 0.36, respectively and indicated that butachlor has a risk to algae in rice paddy, drain and river. On the other hand, TERs of invertebrate in rice paddy, drain and river were 3.6, 50 and 357, respectively, well above 2, indicating no risk to invertebrate. TERs of fish in rice paddy, drain and river were 0.58, 8 and 57, respectively. The TERs for fish indicated that butachlor poses a risk to fish in rice paddy but has no risk to fish in agricultural drain and river. In conclusion, butachlor has a minimal risk to algae in agricultural drain and river exposed from rice drainage but has no risk to invertebrate and fish.

• 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.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.701-710
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• 2002

Studies on the Viability of In Vitro-Matured Bovine Oocytes Vitrified by Microdrop and Straw Method To establish vitrification method for bovine oocytes, mature bovine oocytes were vitrified by microdrop (MD) or straw (Straw) method and the viability of vitrified oocytes with or without cumulus cells (CC) were examined by several methods; a) parthenogenetic activation; b) pronuclear formation after in vitro fertilization (IVF); and c) embryonic development after IVF. The survival rate of vitrified oocytes by MD was significantly higher than by Straw (92.50 vs. 74.19%, p<0.05). Most of the oocytes survived from vitrification using the MD methods. Cleavage and blastocyst development of parthenogenetically activated oocytes were higher in MD (45.05% and 10.81%, respectively; p<0.05)) than those in Straw method (27.17% and 6.52%, respectively; p<0.05). Male and female pronuclear formation of vitrified-thawed oocytes with or without cumulus cells (CC) after IVF were examined, respectively. The survival rate of vitrified oocytes by MD without CC was no difference between MD and Straw (80.368.14% vs. 67.31%). Normal fertilization (2PN) rates were not different among groups (Fresh; 54.55% vs. MD; 42.22% vs. Straw; 37.14%, p>0.05). While no fertilization (<1PN) rates were significantly different between fresh and vitrified-thawed groups (Fresh; 32.47% vs. MD; 57.78% and Straw 62.86%, p<0.05). The polyspermy (3PN) was appeared in the fresh (12.99%), but no appeared in the vitrified-thawed groups. In the without CC, normal fertilization (2PN) rates were significantly different between fresh and vitrified-thawed oocytes (Fresh; 59.38% vs. MD; 17.31% and Straw; 30.43%, p<0.05). Moreover, no fertilization (<1PN) rates were significantly different between fresh and vitrified-thawed groups (Fresh; 23.44% vs. MD; 73.08% and Straw 58.70%, p<0.05). The polyspermy (3PN, >4PN) was appeared not only fresh but vitrified-thawed groups. After IVF, two-cell developmental rates of vitrified oocytes with CC by MD and Straw were significantly low compared to fresh oocytes (Fresh; 81.76% vs. MD; 22.22% and Straw; 11.36%, p<0.05). Blastocyst developmental rates of vitrified oocytes also were significantly low compared to fresh oocytes (Fresh; 28.38 vs. MD; 1.71% and Straw 0%, p<0.05). In the without CC, two-cell developmental rates were no difference between Fresh and MD (27.59% vs. 19.25%, p<0.05), while blastocyst rates were difference between Fresh and MD or Straw (4.31% vs. 0.62% and 0%, respectively; p<0.05). In conclusion, the results indicate that the vitrified bovine oocytes have the ability to develop to the blastocyst stage after IVF.

• The Korean Journal of Malacology
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• v.27 no.2
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• pp.115-119
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• 2011

To establish technical development for artificial seed production, growth and survival for early young spats of the hard clam, Meretrix petechialis, were investigated by breeding methods. Adult clams were collected at Hasa-ri, Baeksu-eup, Yeonggwang-gun, Jeollanam-do on July 13, 2010, and then transported to the indoor aquarium at the laboratory. Eggs which were taken from mother clams, were inseminated, and after they were fertilized in the aquarium, 60 million bottom-clinging spats ($198{\pm}12{\mu}m$ in shell length) were produced and bred. The breeding experiments were carried out from July 16 to October 4, 2010 for 80 days. The methods of sand box, sand bottom circulation filter, inclosing net, floor were used for the breeding experiments, and the experimental condition of sea water temperature for larvae were at 25, 28, 31, $34^{\circ}C$. Four marine cultured food organisms were used for this study as follows: Isochrysis galbana, Chaetoceros gracilis, Phaeodactylum tricornutum, Tetraselmis tetrathele. According to the experimental conditions, experimental groups of the spats in the early stage were investigated the growth rate and the survival. As the result, the method of the inclosing net section was the fastest (grew up to $2.64{\pm}0.59{\mu}m$ in shell length), followed by sandbox ($2.59{\pm}0.64{\mu}m$, bottom circulating filter ($2.56{\pm}0.52{\mu}m$), and floor ($2.52{\pm}0.56{\mu}m$). The survival was the highest in the experimental condition of sandbox (35.9%), followed by floor (34.6%), bottom circulating filter (29.5%), and inclosing net (9.3%). Eexperimental condition of water temperature of $34^{\circ}C$ showed the fastest growth rate (grew up to $2.70{\pm}0.76{\mu}m$ in shell length), and showed the latest growth rate (grew up to $2.45{\pm}0.41{\mu}m$ in shell length) at $25^{\circ}C$. The survival (%) was the highest under the water temperature conditions at $31^{\circ}C$, and showed the lowest (14.2%) at $34.^{\circ}C$. The growth rate of the experimental group fed the mixture live food was the fastest with shell length $2.52{\pm}0.66{\mu}m$, and that of experimental group fed P. tricornutum showed the latest (grew up to $2.29{\pm}0.43{\mu}m$ in shell length). The survival was the highest (36.9%) under the experiment condition fed mixture live food and experimental group fed T. tetrathele showed the lowest rate (16.2%).