• Journal of the Korean Society of Marine Environment & Safety
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• v.26 no.5
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• pp.514-522
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• 2020

To understand the ecotoxicological differences between representative Skeletonema sp. and Dunaliella tertiolecta, both producers as international standard test species for marine ecotoxicity testing, we compared each standard test method, and comparatively analyzed the suitability of the species for environmental assessment and their sensitivity to various test substances. Although most of the test conditions were the same in each method, there were differences in limitation of pH changing and the initial inoculation density in the validation criteria, which is supposed to originate from the low growth rate of D. tertiolecta. In terms of suitability, both species showed consistency in test performance by repeatedly meeting the validation criteria required by the standard test methods. The salinity ranges available for testing were 20 and 10 psu for Skeletonema sp. and D. tertioelecta, respectively. Finally, regarding sensitivity, the toxicity sensitivity of Skeletonema sp. was relatively higher than that of D. tertiolecta for the reference toxicant, actual polluted water discharged (ballast water), and other chemicals. This implies that using at least two species of microalgae from different classification groups could help increase the reliability and objectivity of test results in the performance of marine ecotoxicity tests using producers.

• Environmental Analysis Health and Toxicology
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• v.25 no.1
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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.

• Korean Journal of Environmental Biology
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• v.29 no.1
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• pp.23-30
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• 2011

In this study, the Comet assay (evaluation of DNA damage) used the fish hepatocellular carinoma cell, PLHC-1, was tried to the sediment extract obtained from freshwater to understand its applicability as a tool for monitoring sediment toxicity. In parallel, induced EROD (7-ethoxyresorufin- O-deethylase) activity and DNA damage (TEM values) in PLHC-1 cells were measured for establishing the tandem endpoints of the PLHC-1cell test to test the ecotoxicity of sediment. Among several study sites in a small river passed through downtown and industrial park area, one of them, site B, showed a higher level of EROD activity and DNA damage than other sites. It indicates that a tandem endpoints of PLHC-1 cells could be useful tools for assessing the toxicity of sediment. The sensitivity of Comet assay with PLHC-1 cells was a little higher than that with a blood cell of frog tadpoles to the solvent extract of sediment. According to the results, a PLHC-1 cell-Comet assay could be used as a useful tool for evaluating ecotoxicity of the freshwater sediment. In addition, more detailed studies are needed to the contaminated site.

• Journal of Environmental Science International
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• v.20 no.9
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• pp.1087-1093
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• 2011

The mixotrophic marine ciliate Mesodinium rubrum possesses a highly modified algal endosymbiont as a nutrition source for the species. Accordingly, we assumed that the species can reflect the ecotoxicity on marine producer (as phytoplankton) and consumer (as zooplankton) both. A series of experiments were conducted to identify the potential of the species as a standard test species for marine ecotoxicological study. The comparison of species sensitivity on reference toxic materials was made using potassium dichromate for phytoplankton and copper chloride for zooplankton. The ciliate revealed the highest sensitivity on both reference materials among the seven test species including phytoplankton, benthic copepod and rotifer species. The toxicity end point of the species was 72hr-$EC_{50}$=1.52 mg/L (as potassium dichromate) estimated by population growth inhibition (PGI), which is more sensitive than the most sensitive phytoplankton Skeletonema costatum (72hr-$EC_{50}$=3.05 mg/L). As comparison to rotifer, it also revealed higher sensitivity on copper chloride; 72hr-$EC_{50}$=0.38 mg/L for ciliate and 48hr-$EC_{50}$=0.48 mg/L for rotifer. Also, the elutriate toxicity test of various ocean disposal wastes were conducted to identify the potential of ciliate toxicity test application using industrial waste sludges. The toxicity of leather processing waste sludge was highest on the ciliate, followed by dyeing waste sludge and dye production waste sludge as an increasing order of toxicity. 72h-$EC_{50}$ of ciliate PGI test was 1.83% and that of S. costatum 3.84% for leather waste sludge which showed highest toxicity. The toxicity test results also revealed that the highest sensitivity was observed on ciliate species on ocean disposed sludge wastes. Also, ciliate toxicity test well discriminated the degree of toxicity between sludge sources; 72h-$EC_{50}$ values were 1.83% for leather processing waste sludge, 16.75% for dye production waste sludge and 27.75% for textile production waste sludge. Even the laboratory culture methods of the species were not generally established yet, the species has high potential as the standard test species for marine toxicity test in terms of the dual reflection of phyto- and zooplankton toxicity from single test, sensitivity and test replicability.

• Journal of Environmental Science International
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• v.14 no.12
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• pp.1129-1139
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• 2005

A series of experiments was conducted to identify the potential of five phytoplankton species as standard test species for marine ecotoxicological tests. The candidate phytoplankton species are Skeletonema costatum, Heterosigma akashiwo, Prorocentrum micans, Isochrysis galbana, and Tetraselmis suecica. Salinity tolerance and sensitivity on potassium dichromate as a reference material were identified. Toxicity of eleven ocean dumped sewage sludges and four red tide expellent extracts were estimated by the inhibition of population growth rates (PGR) of marine diatom S. costatum, While most species revealed relatively weak tolerance on salinity, T. suecica demonstrated the highest salinity tolerance ranged from $5\~35$ psu and the others $15\~35$ psu. H. akashiwo revealed the highest sensitivity as 72h $IC_{50}$=0.76mg/L and T. suecica the lowest as 72h $IC_{50}$=8.89mg/L on potassium dichromate. Sludge extracts from industrial waste, domestic sewage and livestock farm waste sludge showed high toxicity as 72h $IC_{50}$<$2\%$ and lowest toxicity from filtration bed sludge as 72h $IC_{50}$=$30.50\%$ NOEC (No Observed Effective Concentration) of sludge extract ranged from <$0.4\%$ to $1.6\%$ and this indicated high phytotoxicity of ocean dumped sewage sludge. The test sensitivity of phytoplankton PGR inhibition was much higher than those of marine rotifer Brachionus plicatilis mortality test and bioluminescent inhibition test by marine bacteria Vibrio fischeri, and comparable with the sea urchin (Strongylocentrotus intermedius) fertilization test. As a result the phytotoxicity test using phytoplankton PGR inhibition ($IC_{50}$) must be a useful tool for marine phyto-toxicological evaluation of ocean dumped materials.

• Ocean Science Journal
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• v.40 no.2
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• pp.91-100
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• 2005

Bioassay using the marine bacteria, Vibrio fischeri and rotifer, Brachionus plicatilis, and chemical analyses were conducted to assess the toxicity of the various sewage sludges, one of the major ocean dumped materials in the Yellow Sea of Korea. Sludge elutriates extracted by filtered seawater were used to estimate the ecotoxicity of the sludge. Chemical characterization included the analyses of organic contents, heavy metals, and persistent organic pollutants in sludge. Bacterial bioluminescent inhibition (15 min), rotifer mortality (24 hr) and rotifer population growth inhibition (48 hr) assay were conducted to estimate the sludge toxicity. EC50 15 min (inhibition concentration of bioluminescence after 15 minutes exposed) values by Microtox(R) bioassay clearly revealed different toxicity levels depending on the sludge sources. Highest toxicity for the bacteria was found with the sludge extract from dyeing waste and followed by industrial waste, livestock waste, and leather processing waste. Clear toxic effects on the bacteria were not found in the sludge extract from filtration bed sludge and rural sewage sludge. Consistent with Microtox(R) results, rotifer neonate mortality and population growth inhibition test also showed highest toxicity in dyeing waste and low in filtration bed and rural sewage sludge. High concentrations of persistent organic pollutants (POPs) and heavy metals were measured in the samples from the industrial wastes, leather processing plant waste sludge, and urban sewage sludge. However, there was no significant correlation between pollutant concentration levels and the toxicity values of the sludge. This suggests that the ecotoxicity in addition to the chemical analyses of various sludge samples must be estimated before release of potential harmful waste in the natural environment as part of an ecological risk assessment.

• Journal of the Korean Society of Marine Environment & Safety
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• v.27 no.6
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• pp.744-753
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• 2021

This study suggests ways to improve the standard test method and judgment criterion for the "Eco-toxicity Test" based on the rules and regulations provided in 'performance and qualifying test standards for marine pollution prevention materials and chemicals' in the Republic of Korea. Compared with the relevant references of other countries, this study attempted to find the limitations in the existing standards. As for the growth inhibition test of algae using Skeletonema costatum as an indicator, applying comparative analysis to measure specific growth rates, together with statistical analysis, instead of applying current methods, judged by the appearance of colors from the culture medium was suggested. Considering the exponential growth phase of the test species, the test duration was suggested to be reduced to less than four days. For the test with fish as an indicator, resetting the appropriate body size was suggested to, show consistent sensitivity against chenicals applied during testing. Furthermore, it is necessary to consider the criteria needs, which should be established in reasonable and objective ways. For the testing species, marine rotifer, Brachionus plicatilis could be a replacement for Artemia sp., and a bivalve for fish in the test. To improve the performance effectiveness of the "Eco-toxicity test", it is worth considering the designation of accredited testing institutes, by placing it in the same loop. Thus it is also expected to have a reliable management system in place, having the capacity of cost calculation properly and adjusting testing items if required.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.4
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• pp.2940-2944
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• 2015

Dredged materials for effective use in Hwasun-port pollution analysis, Ni(nickel) in excess of the standard showed that the whole point. However, according to the revised standards effective utilization exceeds the reference value, even if no anthropogenic sources Pearl ratio less than 10% of the average weight of the ecotoxicological tests have passed, and if so it can be used to beach nourishment. Therefore, the average weight ratio of pearl and ecotoxicological tests were conducted by marine environment official test method. Mud to 5.37% of the average weight ratio lower than the reference value of 10% that was investigated. Ecotoxicity test using bioluminescent bacteria relative luminescence inhibition in the 0 to 9.13% with no toxicity was determined using benthic amphipods in the control and the test sample relative to the average survival rate is 0% with no toxicity was determined. Therefore, the revised effective use meets all the criteria that can be used to beach nourishment, so that the suitability was assessed.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.13 no.2
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• pp.106-111
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• 2008

Six standard methods for marine ecotoxicological tests were established(or applicated) using marine decomposer, primary producers and consumers. Development processes referred to the standard methods established by USEPA(United States Environmental Protection Agency), international organizations and European methods. However, the standard test species were selected among the domestic species generally found in the Korean waters and sediments. The test methods provide the culture/maintenance of test species, test methods, reproducibility and quality acceptance criteria etc. A total of nine test species were designated including bioluminescent bacteria(Vibrio fischeri), diatom(Skeletonema costatum), seaweed(Ulva pertusa), rotifer(Brachionus plicatilis), benthic copepod(Tigriopus japonicus), benthic amphipods(Mandibulophoxus mai, Monocorophium acherusicum), and fishes(Oryzias latipes, Paralichthys olivaceus). These test species represent the decomposer, primary producer and consumers in marine trophic system in Korean coastal ecosystems, and we recommend the "battery test" including at least one species from the each trophic level for marine ecotoxicological test.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.13 no.2
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• pp.129-139
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• 2008

Marine ecotoxicological standard method was applicated using marine rotifer and benthic copepod as primary consumer of marine ecosystem. Marine rotifer, Brachionus plicatilis and benthic copepod, Tigriopus japonicus were designated as standard test species with the endpoints of 24hr neonate mortality (24hr $LC_{50}$) and 48hr population growth (48hr $EC_{50}$) for rotifer, and 48hr nauplius mortality (48hr $LC_{50}$) for benthic copepod. Tests method was referred to those of ASTM (American Society for Testing and Materials) with the replacement of test species which are widely distributed in Korean waters. The two species showed a wide tolerance on salinity ($5{\sim}35\;psu$) and can be easily cultured in small space even they were not as sensitive as in the mortality test using the nauplius of marine invertebrates. However, these species revealed the significant "concentrationresponse relationship" tested with ocean disposal wastes, and reproducibility using cadmium as standard reference material between laboratories. Accordingly, these two species have good potential as test species for marine ecotoxicological test species. Also, we strongly recommend that at least one of these species be included in the test species of "battery test" in marine bioassay.