• 약학회지
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• 제37권5호
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• pp.527-531
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• 1993

As a part of chemical study on the bioactive metabolites from marine organisms, we have investigated cytotoxicity using brine shrimp bioassay for each solvent fractions of the marine algae(12 species), marine sponges(3 species), coelenterates(2 species), echinoderms(4 species), marine molluscs(17 species), and ascidians(2 species), respectively. As the results, chloroform extract of Stichopus japonicus (LC$_{50}$ : 274 $\mu\textrm{g}$/ml), ethyl acetate extract of Anthocidaris crassispina(LC$_{50}$ : 121 $\mu\textrm{g}$/ml), n-butanol extract of Unda), ia Pinnatifida (LC$_{50}$ : 178 $\mu\textrm{g}$/ml), and water extract of Thais clavigera (LC$_{50}$ : 61 $\mu\textrm{g}$/ml) displayed the most significant cytotoxic activity against brine shrimp. Among the marine organisms tested, echinoderms and marine molluscs were thought to be the most active Phylums on screening of new bioactive compounds.

• 한국패류학회지
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• 제21권1호
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• pp.25-31
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• 2005

The embryos of marine bivalves have been commonly used in bioassays for the quality assessment of marine environments. Although several standard protocols for developmental bioassay with bivalves have been already proposed, there have been few trials for applying these protocols in environmental assessment, or for developing new protocol with Korean species. So, there is a strong need to establish the standard bioassay protocols using bivalves commonly found in Korean waters. Prior to developing a new protocol, it is essential to know the optimum conditions for the reliable bioassay procedures. Here, we established the purpose of this study to determine the optimum bioassay conditions for successful development of a common mussel, Mytilus galloprovincialis. The conditions considered as critical for developmental bioassay, and determined in this study were; (1) temperature, (2) salinity, and (3) initial density of embryo. The optimal temperature for developmental bioassay of M. galloprovincialis was determined as $15^{\circ}C$. At this temperature, the required time for the embryo to become veliger larva was 48 hr. The acceptable range of salinity for the embryotoxicity test using M. galloprivincialis was from 30 to 35 psu, which was narrower than that of the natural habitat of adult populations. The optimum density of embryo at the beginning of bioassay was 100 embryos/ml. Over this density, the proportion of normally developed larvae decreased significantly. The results obtained in this study will serve as a basis for preparation of the standard bioassay protocol using embryo of M. galloprovincialis.

• 한국패류학회지
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• 제22권2호
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• pp.167-173
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• 2006

The embryos of marine bivalves have been commonly used in bioassays for quality assessments of marine environments. Although several standard protocols for the developmental bioassay of bivalves have been proposed, only a few trials for application of these protocols in environmental assessments or for the development of a new protocol with Korean species have been conducted. As such, there is a strong need to establish standard bioassay protocols with bivalves commonly found in Korean waters. To determine the sensitivity of Mytilus galloprovincialis to establish a standard bioassay, their fertilized eggs were exposed to six metals (Ag, Cd, Cr, Cu, Ni, and Zn). The order of biological impact was Ag > Cu > Ni > Zn > Cr > Cd and their lowest observed effective concentration were 5, 16.4, 25.4, 142, 187 and 1,500${\mu}g/l$, respectively. The proportion of normal larvae appeared to decrease linearly with the logarithm of each toxicant concentration within the tested range. The average values of median effective concentrations $(EC_{50})$ from the triplicate experiments for Ag, Cd, Cr, Cu, Ni, and Zn were 6.8, 1,797, 786, 16.6, 68.1, and 139.2${\mu}g/l$, respectively. There was a more than 100-fold difference in $EC_{50}$ values of Cu and Cd. The value of $EC_{50}$ or median lethal concentration of Cu was within the range observed for other bivalve developmental bioassays. The overall sensitivity of M. galloprovincialis in the present developmental bioassay was also similar to that of other marine organisms commonly used in aquatic bioassays (e.g. sea urchins, oysters). Hence, the bioassay using the embryo of M. galloprovincialis is considered to be a useful tool to monitor and evaluate the quality of marine aquatic environments.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 추계국제학술대회
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• pp.158-158
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• 2003

Bioassays using gametes of sea urchins are widely used in ecotoxicological assessments of marine environments. Since most of sea urchin species in Korean coastal water spawn from spring to autumn, bioassay with them during the winter is impossible. In the course of developing standard methods for bioassays with Korean species, we found a winter-spawning starfish, Asterias amurensis, Since reproductive mode of asteroids is similar to echinoids, the bioassay protocol for sea urchins could be applied similarly to the starfish. Here, we tested and determined several conditions for the acceptability of bioassay with A. amurensis. The least required time for formation of fertilization membrane of fertilized eggs to be easily distinguished from unfertilized ones was 60 min. The threshold of sperm to egg ratio that could make acceptable fertilization rates in controls was 3000. The allowed time for manipulation of sperm after dilution in seawater was at most 3 hr. The optimal exposure time of sperms when the response against toxicant solution was relatively stable was in the range of 20-60 min. The tolerance range of sperms to the salinity of test solution was 26-38 psu. The sensitivity of A. amurensis sperm was intermediate among marine organisms commonly used in aquatic toxicity tests. The sperm bioassay with A. amurensis can be satisfactorily applied to toxicity assessments of marine environments.

• Environmental Analysis Health and Toxicology
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• 제25권1호
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• pp.19-26
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• 2010

Discharged materials from the point or non-point source are released into the sea, and as the results, marine environment is directly affected. We must estimate the impacts of contaminants to marine pollution rapidly and accurately. Therefore, it is needed on early warning system for appreciating marine environmental impacts, and required a bioassay to evaluate abnormal changes. A bioassay test was developed to examine the effects of heavy metal contaminants on the early life stages of the marine annimals. We have studied the effects of metals on early development of a sea urchin species, Strongylocentrotus intermedius. S. intermedius embryos were tested with six metals (Cu, Ag, Zn, Cd, Cr, Ni) and showed the highest sensitivity to Cu as well as the lowest sensitivity to Cd. The order of biological impact for metals was Cu>Ag>Ni>Zn>Cr>Cd. In accordance with the results, sea urchins embryos can provide biological criteria for seawater quality assessment. The sensitivity of developmental bioassay whith S. intermedius is at intermediate level among marine organisms commonly used in aquatic bioassays. And this sea urchin can be routinely employed as a test organism for ecotoxicity assays.

• 해양환경안전학회지
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• 제12권2호
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• pp.99-106
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• 2006

해양환경에서 오염물질의 거동은 화학적 특성뿐 아니라 부유물질 표면의 정전하, pH 및 염분과 같은 환경 인자에 크게 영향을 받을 것이다. 이 연구는 해산어 양식장에서 빈번히 사용하고 있는 항생제인 옥소린산의 거동이 여러 종류의 부유물질, 염분 및 pH에 따라 어떻게 변동하는가를 HPLC에 의한 화학적 분석과 bioassay 분석으로 검토하였다. 부유물질의 농도가 증가할수록 옥소린산은 HPLC 분석에 의해서도 저농도로 검출되었고, 생물활성에 의한 bioassay 분석으로도 현저히 감소하는 것으로 나타났다. 이 결과는 부유물질이 옥소린산을 흡착하여 수서환경에서 제거하는 역할을 한다는 것을 알 수 있다. 그리고 옥소린산은 염분 40%o과 pH 7에서 bioassay 분석에 의해 생물활성이 약간 저해되지만 HPLC 분석은 조금 다른 양상을 보였다.

• 한국패류학회지
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• 제30권3호
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• pp.177-187
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• 2014

Mussels have been commonly used in bioassay for quality assessments of environment. Moreover, several standard protocols for the developmental bioassay of bivalves have been proposed. In this study, the EC50 of polycyclic aromatic hydrocarbons (PAHs) was determined using mussel, Mytilus galloprovincialis embryonic developmental bioassay. To determine the sensitivity of M. galloprovincialis, their fertilized eggs were exposed to five PAHs (2-metylnaphthalene, fluorene, dibenzothiophene, phenanthrene, pyrene). The EC50 of 2-metylnaphthalene, fluorene, dibenzothiophene, phenanthrene, and pyrene were 232, 273, 67.9, 43.2, and $33.1{\mu}g/L$, respectively. The overall sensitivity of M. galloprovincialis in the present developmental bioassay was similar to or more sensitive than that of other marine organisms commonly used in aquatic bioassays. The results of this study could be provide with fundamental data of setting standard for protection of marine life and or can use prediction the aquatic toxicity of PAHs.

• 한국환경과학회지
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• 제14권11호
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• pp.1071-1079
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• 2005

Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.

• 생명과학회지
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• 제9권6호
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• pp.653-658
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• 1999

A novel marine microorganism, Vibrio sp. YE-101, was isolated from pufferfish and investigated for its ability to synthesize tetrodotoxin (TTX). Various strains isolated from the intestine of pufferfish were grown on TCBS agar plate, and then cultured on Ocean Research Institute (ORI) medium supplemented with 3% NaCl at 23$^{\circ}C$ for 3days. The cells were harvested, disrupted, fractionated by Bio-Gel P-2 column chromatography and then TTX-producing strain, Vibrio sp. YE-101, was identified using mouse bioassay. The isolated TTX from Vibrio sp. YE-101 was also analyzed and identified by HPLC and gas chromatography-mass spectrometer (GC-MS). The mass fragmentation of trimethylsilyl derivatives of C9-base of TTX from Vibrio sp. YE-101 was interpreted and the pattern of fragmentation was same with that of authentic standard. The purfied TTX was also positive to the mouse bioassay, which clearly represents that Vibrio sp. YE-101 can synthesize TTX.

• 해양환경안전학회지
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• 제28권4호
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• pp.459-468
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• 2022

천수만과 태안해역의 제한영양염을 평가하기 위해 장기자료 분석과 생물검정실험을 진행하였다. 우선 잠재적인 제한영양염을 평가하기 위해 국가수질측정망에서 제공되는 2004~2016년 동안의 장기 영양염 자료를 이용하였다. 장기자료의 DIN/DIP를 분석한 결과 대부분 16이하로 N 제한이 우세하였지만 N, P, Si의 농도비를 이용한 분석에서는 하계와 추계에는 N 제한이 우세하였고, 동계와 춘계에는 해역에 따라 일부 Si 제한을 보이거나 또는 제한이 나타나지 않았다. 생물검정실험 시 채집된 현장수의 영양염 분석결과, DIN/DIP는 3월과 5월에 모든 정점에서 P 제한을 나타냈고, 7월과 10월에는 N 제한이 우세하였다. N, P, Si의 농도비를 이용한 분석에서 3월과 5월은 P와 Si 제한을 보이거나 제한영양염이 나타나지 않은 정점이 존재하였으나 7월과 10월에는 N 제한이 우세하였다. 실질적인 제한영양염을 평가하기 위해 수행된 생물검정실험 결과 3월에는 특정 제한영양염이 나타나지 않았으나, 5월, 7월 10월에는 NH₄⁺와 NO₃^-가 반응을 보임으로서 이 시기에는 N이 식물플랑크톤 성장에 직접 관여하는 실질적인 제한영양염임을 확인하였다.