• Korean Journal of Ichthyology
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• v.22 no.1
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• pp.25-33
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• 2010

Effects of different salinity levels on spawning performance, embryonic development and early viability of a euryhaline medaka species, Oryzias dancena, were examined. O. dancena were able to spawn eggs in a wide range of salinity from 0 to 70$^{\circ}/_{\circ\circ}$, however, the spawning frequency was lowered in complete freshwater (0$^{\circ}/_{\circ\circ}$) and in highly salted water (70$^{\circ}/_{\circ\circ}$). Fertilization success was negatively affected when the environmental salinity was higher than the salt concentration found in normal seawater. Embryonic viability and hatching success were also inversely related with the salinity levels. Typical abnormality was observed in developing embryos incubated at high salinities (30, 45 and 60$^{\circ}/_{\circ\circ}$). In addition, the time to hatch was significantly delayed with increasing salinities: peak hatching occurred at 12~14 days post fertilization (dpf) in freshwater and at least at 17 to 18 dpf in 60$^{\circ}/_{\circ\circ}$. Mean survival rates of the hatched larvae up to 7 days post hatching (dph) were at least 97% in salinity levels ranging from 0 to 30$^{\circ}/_{\circ\circ}$. However, larvae reared in 45 and 60$^{\circ}/_{\circ\circ}$ experienced significant mortality, especially in the early phase, resulting in only 75% and 64% survival rates up to 7 dph, respectively.

• Korean Journal of Environmental Biology
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• v.27 no.4
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• pp.391-396
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• 2009

The biological effects of carcinogenic polycyclic aromatic hydrocarbons (cPAHs) including benzo[a]pyrene (BaP), dibenzo[a,h]anthracene (DBA), benzo[a]anthracene (BaA), benzo[b] fluoranthene (BbF), benzo[k]fluoranthene (BkF), and indeno[1,2,3-c, d]pyrene (InP) on transcriptomic changes were determined in the liver of Oryzias latipes. Differentially expressed genes (DEGs) by binary exposure to cPAHs (BaP+BaA, BaP+BbF, BaP+BkF, BaP+DbA, BaP+InP) were screened by annealing control primers-based polymerase chain reaction followed by sequence analysis and BLAST searching. The results showed that four DEGs were commonly expressed by cPAHs and they were identified as ribosomal protein S4, coagulation factor II, elongation factor 1 beta, and a predicted protein similar to human immunodeficiency virus type I enhancer binding protein 3. This finding suggests that binary exposure to cPAHs interferes protein synthesis required for fundamental liver functions in fish.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.06a
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• pp.279-285
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• 2005

Endocrine disrupting effects of four pharmaceutical products were evaluated with fish. The test pharmaceuticals, i.e., sulfamethoxazole, sulfamethazine, oxytetracycline and tetracycline have been often detected in aquatic environment, but their ecological hazard on receptors of various trophic levels has seldom been evaluated. In the present study, we conducted acute toxicity assays with a fish, Japanese medake (Oryziα lαtipes). The vitellogenin induced in female fish normally, but a endocrine disrupting chemical could give effects even male fish. We have tried 4 pharmaceutic chemicals to find out the endocrine disrupting effects. Sulfamethoxazole 1, 0.5 ppm induced vitellogenin even at male Japanese medaka. Sulfamethazine 10, 5, 1 ppm could induced vitellogenin at male fish. Oxytetracycline 10, 5, 1ppm could induced vitellogenin With the fish. Tetracycline 10, 5 ppm could induced vitellogenin at male fish. Some pharmaceuticals such as sulfamethoxazole, sulfamethazine, oxytetracycline and tetracycline could give effects to male Oryzias latipes. They could induced vitellogenin under exposure range 0.5 ${\sim}$ 10 ppm of chemicals at male Oryzias latipes.

• Environmental Analysis Health and Toxicology
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• v.20 no.4 s.51
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• pp.359-363
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• 2005

ESS (Embryo and sac-try stage) 독성 시험에서 시험어종으로서의 국내토착종인 왜몰개 (Aphyocypris chinensis)의 적용성을 평가하기 위하여 아연(Zn)을 사용하여 국제적인 추천시험 어종인 송사리(Oryzais latipes)와 감수성을 비교하였다. 시험기간은 대조군에서 아사가 관찰되는 시기 즉, 왜몰개는 수정 후 8일, 송사리는 수정 후 16일로 하였으며, 시험기간 동안 수정란의 부화율, 수정란 및 난황단계의 치어(sar-fry)의 사망률, 형태적인 발달, 치어의 성장 등을 관찰 및 측정하였다. 두 종 모두 수정란의 생존율에 아연의 영향을 받았으며, LOEC는 모두 14.5 mg/L이었다 난황단계 치어의 사망률을 관찰한 결과, 왜몰개는 1.4mg/L부터 급격히 증가된 반면에 송사리는 14.5mg/L에서 $100\%$사망률이 관찰되었다. 시험물질에 노출된 왜몰개와 송사리 모두 척추변형이 관찰되었으며, 체장을 측정한 결과는 왜몰개가 송사리에 비해 민감하게 반응하는 것으로 나타났다. 위와 같은 결과들은 종합해 볼 때, ESS독성시험에서 왜몰개가 대체 시험어종으로서 사용될 수 있는 가능성을 보여주었다

• Environmental Analysis Health and Toxicology
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• v.22 no.4
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• pp.305-312
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• 2007

환경오염을 신속하게 모니터링하기 위한 생물지표의 개발은 증가하고 있는 오염의 심각성에 비추어 매우 중요한 과제로 여겨지고 있다. 본 연구에서는 독성물질처리에 의하여 선택적으로 발현이 조절되는 단백질의 동정을 통하여 독성물질에 대한 단백질 생물지표를 발굴하고자 시도하였다. 즉, 송사리(Oryzias latipes)를 유기인계 살충제인 다이아지논(diazinon)에 0, 0.1, 1, 5 mg/L 농도로 24시간 노출시킨 후, 머리와 몸통부분으로 나누어 단백질 발현패턴을 분석하였다. 본 시스템에서 다이아지논 처리에 의하여 유의적으로 발현이 증가된 단백질로서 alpha-tubulin, ribonuclease pancreatic precursor, protein hfq 등을 동정하였으며, 이 가운데 alpha-tubulin과 $hsp90{\beta}$의 발현이 다이아지논 농도에 의존적으로 증가하는 것을 semi-quantitative RT-PCR방법으로 확인하였다. 이와 같이 다이아지논 처리에 특이적으로 발현이 증가된 송사리 단백질들은 노출평가를 위한 생물지표로서 개발에 응용될 수 있을 것으로 평가된다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.2
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• pp.160-168
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• 2017

Optimizing the conditions for stem cell culture is an essential prerequisite for the efficient utilization of stem cells. In the culture of fish embryonic stem cells (ESCs) or ESC-like cells, embryo extracts are important for stable growth, but there is no rule for determining the developmental stage of the embryos used to obtain extracts. Therefore, this study investigated the effects of the developmental stage of extract donor embryos on the culture of Oryzias dancena ESC-like cells. O. dancena ESC-like cells were cultured in different media containing each of four types of embryo extract depending on the developmental stage of the extract donor embryos. Growth, morphology, colony-forming ability, alkaline phosphatase (AP) activity, and embryoid body (EB) formation of the cells were investigated. While the developmental stage of the extract donor embryos did not influence the growth, morphology, AP activity, or EB formation of ESC-like cells, colony-forming ability was affected and the pattern of the effects differed completely between the two ESC-like cells investigated. These results suggest that the developmental stage of extract donor embryos should be selected carefully for the culture of ESC-like cells, according to the research purpose and type of cell line.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.2
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• pp.270-283
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• 2014

본 연구는 온도 변화에 따른 해산송사리, Oryzias dancena의 마취 효과 및 생리적 스트레스 반응을 평가하고, 고온 및 저온에서 최적의 마취 수온을 구명하였다. 해산송사리에서 고온 실험($36{\sim}42^{\circ}C$)과 저온 실험($4{\sim}10^{\circ}C$)을 통해 마취 효과를 조사하였으며, 각 마취수온에서 전어체의 cortisol과 glucose를 측정하였다. 각 수온 마취 실험 후, 전개체는 모두 생존하였으며, 해산송사리의 마취시간은 고온 실험에서 고온 일수록, 저온 실험에서는 저온 일수록 유의적으로 빨랐다(P<0.05). 회복 시간은 유의하게 고온 실험에서 수온이 감소할수록, 저온 실험에서는 수온이 증가할수록 유의적으로 느렸다(P<0.05). 아가미 운동수는, 고온 실험에서 수온이 증가함에 따라 빨랐으며, 저온 실험에서는 수온이 감소할때 빨랐다(P<0.05). 수온이 $38^{\circ}C$ 나 $8^{\circ}C$ 일때의 마취 조건에서 실험직후 전어체 cortisol이 최대치로, 실험후 6시간까지 점진적으로 감소한 반면, 전어체 glucose는 실험후 1시간에 최고치를 보이고 실험후 2시간까지는 감소하였다. 기존의 마취제를 쓰지 않은 본 연구로 인해, 해산송사리의 여타 연구에서 샘플 및 사용자가 보다 더 안전하고 용이한 샘플 취급이 가능할 것으로 사료된다.

• Environmental Analysis Health and Toxicology
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• v.22 no.2 s.57
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• pp.171-175
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• 2007

Indigenous species means a species that is likely, due to historical presence, to occur at a specified site for some portion of its life span. Therefore, indigenous species can be useful as an indicator to assess environmental risk caused by hazardous chemicals in a specific site. So far a few toxicity studies using freshwater species which are indigenous to Korea have been carried out. In this study, a freshwater shrimp (Neocardina denticulata) indigenous to Korea was used for acute toxicity test of heavy metals. Neocardina denticulata were exposed to cadmium chloride $(CdCl_2)$, copper chloride $(CuCl_2)$ and zinc chloride $(ZnCl_2)$ using automatic flow-through system for 96 hours. The 96h LC50s were calculated as 0.043 $(0.042{\sim}0.045)mg\;CdCl_2/L,\;0.104(0.098{\sim}0.113)mg\; CuCl_2/L\;and\;2.021\;(1.633{\sim}2.594)mg\;ZnCl_2/L$. When compaired with some international standard species such as medaka(Oryzias latipes), Neocardina denticulata had high sensitivity. Therefore, this study suggested that Neocardina denticulata have possibilities for a sensitive test species to test heavy metal toxicity in aqua-system.

• Environmental Analysis Health and Toxicology
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• v.24 no.4
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• pp.321-332
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• 2009

본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2002.10a
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• pp.156-156
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• 2002

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.