• Fisheries and Aquatic Sciences
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• v.17 no.4
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• pp.461-469
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• 2014

Polychlorinated biphenyls (PCBs) are persistent pollutants in aquatic environments, often causing the decline or disappearance of wild populations. In this study, we used a random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of olive flounder embryo and larval stages of exposure to Aroclor 1254 at concentrations of 1, 5, 10, 20, and $40{\mu}g/L$. We compared RAPD fingerprints of exposed and non-exposed samples. Polymorphisms were revealed as the presence and/or absence of DNA fragments between the two samples. A dose-dependent increase in the number of polymorphic bands was observed with Aroclor 1254 treatment. Also, RAPD profiles of animals exposed to Aroclor 1254 exhibited an increase in the frequency values (FV) compared to the control. A phenogram constructed using neighbor-joining method indicated that genomic template stability in developing embryo and larval stages was significantly affected at ${\geq}5{\mu}g/L$. This study suggested that DNA polymorphisms detected by RAPD analysis could be used as an investigative tool for environmental toxicology and as a useful biomarker in early life stages for the detection of potential genotoxicants.

• Korean Journal of Environment and Ecology
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• v.30 no.4
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• pp.705-711
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• 2016

In this experiment, embryos of Asian toad, Bufo gargarizans, were investigated to evaluate toxicity of chemicals along FETAX(Frog Embryo Teratogenesis Assay-Xenopus) protocol. Asian toad, Bufo gargarizans, embryos incubated and investigation of Zn and Benomyl effect by probit analysis. As a result, depends on the concentrations of Zn and Benomyl, mortality and malformation rates were increases and larval body length were decreased. The teratogenic concentration($EC_{50}$) of Zn and Benomyl were 2.3, $1.0mg/{\ell}$, respectively and the embryo lethal concentration($LC_{50}$) Zn and Benomyl were 10.3, 6.9, respectively. The teratogenic index(TI) were 4.4 in Zn and 6.7 in Benomyl, thus showed teratogenicity in embryonic development of B. gargarizans. These results reveal that Zn and Benomyl in this experiment suppressed the development of embryos at relatively low concentration. Much of B. gargarizans embryos can be secured, and easy to incubate. In addition, mortality, malformation ratios, malformation patterns and growth rates are similar to the results from the other assay systems. Therefore, the B. gargarizans embryo teratogenesis assay system could be a useful tool to evaluate toxicity of pollutants in environment.

• Environmental Analysis Health and Toxicology
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• v.15 no.3
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• pp.61-67
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• 2000

Toxic lesions of styrene in the Japanese Medaka (Oryzias latipes) were compared with those of styrene oxide, the active metabolite of styrene, using embryo-larval assays. The developmental stages of Japanese Medaka (Oryzias latipes) treated with both chemicals were not altered and progressed normally. However, styrene oxide was more toxic than styrene in terms of causing death and lesions . High concentrations of styrene (higher than 4.9 ppm) and styrene oxide (higher than 2.4 ppm), resulting in more than 50% mortality, caused similar lesions of cardiovascular system, craniofacial bone formation and spinal deformities, although a number of lesions were not observed by both chemicals . In the group treated with styrene, eyeball sizes and intereye distances were reduced, while, in the group treated with styrene oxide , the eyes and eye cups were not developed and two eyes were sometimes fused. In addition, styrene oxide caused the lesion which involved the posterior brain and brain stem were herniated through the spinal cord . The noticeable difference of toxic symptoms between these two chemicals was the time of onset. Toxicities of cardiovascular system and craniofacial bone formation appeared on day 3 of development in styrene oxide treated group, but, styrene treated group staned to show hemorrhages on day 3 and the craniofacial malformation were appeared on day 5, These differences between two chemicals may be due to the metabolism of styrene to styrene oxide, the reactive intermediate.

• Korean Journal of Environment and Ecology
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• v.33 no.2
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• pp.178-186
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• 2019

This study used the probit analysis to evaluate the toxicity of three chemicals - benomyl (Germicide), carbofuran (insecticide), and thiobencarb (herbicide) - with the FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol using the incubated embryos of tree frog, Hyla japonica. The results showed that the larval body length decreased while the mortality and malformation rates increased as the concentrations of benomyl, carbofuran, and thiobencarb increased. The teratogenic concentration ($EC_{50}$) of benomyl, carbofuran, and thiobencarb were 1.00, 0.58, 4.75 mg/L, respectively, indicating that the malformation of larvae was the most sensitive to carbofuran. The embryo lethal concentration ($LC_{50}$) was 7.04, 28.71, and 16.12mg/L, respectively, indicating that benomyl showed the lowest embryo lethal concentration. The teratogenic index (TI) was 7.04 in Benomyl, 49.50 in Carbofuran, and 3.39 in Thiobencarb, indicating that the TI values were above 1.5, which is the criterion of teratogenicity, for all three chemicals. All three pesticides examined by this study were considered to be the most teratogenic substances, and the carbofuran was the most potent teratogen.

• Korean Journal of Environment and Ecology
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• v.26 no.5
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• pp.675-681
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• 2012

In this experiment, I investigated toxicity evaluation of chemicals using domestic frog embryos, along FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. I investigated $Cu^{2+}$ and Tebuconazole effect on the tree frog, Hyla japonica, embryos by probit analysis. Mortality and malformation rates increased and larval body length decreased depending on the concentrations of $Cu^{2+}$ and Tebuconazole. The teratogenic concentration ($EC_{50}$) of $Cu^{2+}$ and Tebuconazole were 0.05, 5.0mg/${\ell}$, respectively and the embryo lethal concentration ($LC_{50}$) of $Cu^{2+}$ and Tebuconazole were 0.16, 38.5, respectively. The teratogenic index (TI) appeared 3.0 in $Cu^{2+}$ and 7.7 in Tebuconazole, which showed teratogenicity in embryonic development of Hyla japonica. These results reveal that $Cu^{2+}$ and Tebuconazole in this experiment suppressed the development of embryos at relatively low concentration. Much of Hyla japonica embryos can be secured, and easy to incubate. In addition, mortality, malformation ratios, malformation patterns and growth rates are similar to the results from the other assay systems. Therefore, the Hyla japonica embryo teratogenesis assay system could be a useful tool to evaluate toxicity of pollutants in environment.

• Korean Journal of Environment and Ecology
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• v.34 no.3
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• pp.207-215
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• 2020

In this experiment, investigated toxicity evaluation of chemicals using Asian toad embryos, along FETAX(Frog Embryo Teratogenesis Assay-Xenopus) protocol. Asian toad, Bufo gargarizans embryo incubated and investigation of Benomyl(Germicide), Carbofuran(Insecticide) and Thiobencarb(Herbicide) effect by probit analysis. As a result, depends on the concentrations of Benomyl, Carbofuran and Thiobencarb, along mortality and malformation rates were increases and larval body length were decreased. The teratogenic concentration(EC₅₀) of Benomyl, Carbofuran and Thiobencarb were 1.03, 8.74, 4.98mg/ℓ, respectively. And when exposed to Benomyl, larvae responded most sensitively to malformations. Embryo lethal concentration (LC₅₀) Benomyl, Carbofuran and Thiobencarb were 7.26, 560.72, 16.87mg/ℓ, respectively. And Benomyl were at the lowest concentration of lethal the embryos. The teratogenic index(TI) were 7.05 in Benomyl, 64.16 in Carbofuran and 3.39 in Thiobencarb, thus TI values were above 1.5, which is the criterion of teratogenicity. Three of the pesticides used in this study were considered to be a teratogenic substances and Carbofuran was the most potent teratogen. And more specific researches are needed to investigate the effects of pesticides on the embryo development of toads and amphibians and their mechanism.

• Korean Journal of Environmental Biology
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• v.39 no.3
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• pp.311-318
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• 2021

In this experiment, we investigated the toxicity of tebuconazole (fungicide) using domestic frog embryos, along the FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. Bufo gargarizans, Hyla japonica, and Pelophylax nigromaculatus embryos were incubated, and investigation of the tebuconazole effect was performed by the probit analysis. As a result, depending on the concentrations of tebuconazole, the mortality and malformation rates were increased and larval body length was decreased. The teratogenic concentrations (EC₅₀) of tebuconazole were 34.4mg L^-1, 10.6mg L^-1, and 14.9mg L^-1, respectively, and the embryo lethal concentrations(LC₅₀) of tebuconazole were 74.7 mg L^-1, 38.5 mg L^-1, and 39.1 mg L^-1, respectively. The teratogenic index (TI) valuesof tebuconazole were 2.19, 3.58, and 2.65; thus, it showed teratogenicity in embryonic development of these three frogs. These results revealed that in this experiment, tebuconazole suppressed the development of embryos at a relatively low concentration. In addition, mortality, malformation ratios, malformation patterns, and growth rates were similar to the results from the other assay systems. Therefore, tebuconazole was thought to have an effect on the embryo development of domestic frogs. In future, it will be necessary to identify species specificity in order to the clarify the causes of differences in mortality, malformation rate, and malformation patterns depending on the species.

• Korean Journal of Environmental Biology
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• v.30 no.3
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• pp.231-237
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• 2012

Toxicity of $Ni^{2+}$ and Tebuconazole were investigated via FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol using domestic frog embryos. Embryos of Black-spotted pond frog, Rana nigromaculata, were incubated and toxic effects of $Ni^{2+}$ and Tebuconazole were investigated by probit analysis. As a result, mortality and malformation rates were increased and larval body length was decreased in a dose dependant manner of $Ni^{2+}$ and Tebuconazole. The half maximal effective concentration ($EC_{50}$) of $Ni^{2+}$ and Tebuconazole were 0.07, $12.7mg\;L^{-1}$, respectively and the half maximal lethal concentration ($LC_{50}$) of $Ni^{2+}$ and Tebuconazole were 4.2, 39.1, respectively. The teratogenic index (TI) were 61.4 in $Ni^{2+}$ and 3.1 in Tebuconazole, respectively. These results reveals that $Ni^{2+}$ and Tebuconazole suppress the development of Black-spotted pond frog embryos at the low concentration as showing teratogenic effects in other assay system. Therefore, teratogen assay system using the Rana nigromaculata embryos could be useful as a tool to evaluate the toxicity of the pollutants in environment.

• Molecules and Cells
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• v.27 no.5
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• pp.583-589
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• 2009

GTP cyclohydrolase I (GTPCH) is a key enzyme in the de novo synthesis of tetrahydrobiopterin. Previously, the Drosophila melanogaster GTPCH gene has been shown to be expressed from two different promoters (P1 and P2). In our study, the 5'-flanking DNA regions required for P1 and P2 promoter activities were characterized using transient expression assay. The DNA regions between -98 and +31, and between -73 and +35 are required for efficient P1 and P2 promoter activities, respectively. The regions between -98 and -56 and between -73 and -41 may contain critical elements required for the expression of GTPCH in Drosophila. By aligning the nucleotide sequences in the P1 and P2 promoter regions of the Drosophila melanogaster and Drosophila virilrs GTPCH genes, several conserved elements including palindromic sequences in the regions critical for P1 and P2 promoter activities were identified. Western blot analysis of transgenic flies transformed using P1 or P2 promoter-lacZ fusion plasmids further revealed that P1 promoter expression is restricted to the late pupae and adult developmental stages but that the P2 promoter driven expression of GTPCH is constitutive throughout fly development. In addition, X-gal staining of the embryos and imaginal discs of transgenic flies suggests that the P2 promoter is active from stage 13 of embryo and is generally active in most regions of the imaginal discs at the larval stages.

• Korean Journal of Environmental Biology
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• v.35 no.4
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• pp.706-714
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• 2017

An investigation of the effects of Pb for domestic anuran embryos were evaluated with the Frog Embryo Teratogenesis Assay; Xenopus (FETAX). Depending on the species, the difference between the embryo size and the embryonic development time was determined. As a result, mortality and malformation rates were increased, malformation patterns were changed and larval body length were decreased in a dose dependent manner of the Pb. The half maximal lethal concentration ($LC_{50}$) of the Bufo gargarizans, Hyla japonica, Rana nigromaculata and Bombina orientalis were 0.58, 0.49, 0.52, $0.54mg\;L^{-1}$, respectively. The half maximal effective concentration ($EC_{50}$) of the Bufo gargarizans, Hyla japonica, Rana nigromaculata and Bombina orientalis were 0.35, 0.74, 0.30, $0.29mg\;L^{-1}$, respectively. The teratogenic index (TI) were 1.66 in the Bufo gargarizans, 1.81 in the Hyla japonica, 1.73 in the Rana nigromaculata and 1.86 in the Bombina orientalis, respectively. Therefore, the Pb seems likely to have a teratogenic effect in all four species' embryonic development. The Bombina orientalis was the most sensitive to the Pb. This means that the difference between the different species, even if they have all been exposed to the same concentration of pollutants depending on the species. The result above show that the Pb acts as a teratogenic agent in the development of the four domestic frog species.