Muscle Intoxication of 'Gukmeri-bok' Puffer (Takifugu vermicularis radiatus) by Freezing and Thawing

We here report that the muscle of ‘gukmeri-bok’ puffer fish (Takifugu vermicularis radiatus) is intoxicated after freezing and thawing processes even though it is not toxic when it is fresh. This study was carried out to investigate the effect of different freezing and thawing conditions as well as the effect of the presence of skin or viscera on the intoxication of muscle of ‘gukmeri-bok’ puffer during freezing and thawing process. As a result of this investigation, thawing condition, but not freezing conditions or periods played an important role in muscle intoxication. Tetrodotoxin secretory grands exist in the skin of toxic puffer fish, the skin was more responsible for the muscles intoxication than the viscera during the freezing-thawing process. In other words, no toxicity was measured in the muscle of skinned-frozen specimens even when thawed. According to this result, it is recommended that the skin and viscera must be removed before being frozen for edible purpose. Otherwise, when a whole fish should be frozen, alternative half-thawing and removal of skin from frozen specimens is recommended.22Nkc

Effects of Sea-Nine 211 and tributyltin on microsomal monooxygenase system of clam, Pseudocardium sachalinensis

Sea-Nine 211 (4,5-dichloro-2-n-octyl-3(2H) isothiazolone) was injected to clam, Pseudocardium sachalinensis, that inhabitate nothern part of Kangwon Province, Korea. Survival rate of the clam and xenobiotics metabolizing enzyme activities in digestive gland were measured during the 4 day-exposure period. The results were compared with those of TBT exposed clam. There were no mortality of clam in the solvent (DMSO) control group and the three Sea-Nine 211 exposure groups (5, 25, 50 mg/kg body weight), while the clam exposed to 1, 2, 5 mg/kg TBT chloride (TBTC) demonstrated 70, 30, 0% survival rate, respectively. The Sea-Nine 211 exposure group showed a tendency of CYP450 induction according to the exposure duration, on the other hand, CYP450 content was decreased in the TBT exposure group. NADPH cytochrome P450 reductase activity slightly increased according to the exposure duration in the Sea-Nine 211 exposure group, while TBTC inhibit its activity as CYP450 content. There was no significant change of cytochrome b5 reductase activity in the clam exposed to Sea-Nine 211. In the TBTC exposure group, its activity increased in early exposure period and then significantly decreased the rest of exposure period. CYP1A1 dependant EROD activities in both the Sea-Nine 211 and the TBTC exposure groups showed no significant differences compared to those of the solvent control group. CYP1A2 dependant MROD activity was significantly induced at the end of exposure, but no significant difference was obtained in the TBTC exposure group. CYP2B1/2 dependant PROD activities in both the Sea-Nine 211 and the TBTC exposure groups showed no significant differences compared to those of the solvent control group. GST activity in the clam exposed to Sea-Nine 211 demonstrated less than those of the solvent control group in the early exposure period, and then recovered at the end of exposure, while its activity was significantly different at the end of exposure in the TBTC exposure1

In vitro Response of the MPO System of the Clam, Coelomactra antiquata with Exposure to Cytochrome P450 Inducers

강원 북부 연안에 많은 명주조개 (Coelomactra antiquata)의 중장선으로 미크로좀을 만들고, cytochrome P45O (CYP)의 유도제로 알려진 phenobarbital (PB), 3-methylcholanthrene (MC) 및 β -naphthoflavone (BNF)을 DMSO에 녹여 함께 30 ∘C 의 수조 상에서 in vitro로 배양하면서 약물대사효소계의 CYP 함량과 7-othoxyresorufin-O-deethylase (EROD) 활성의 변화를 경시적으로 조사하였다. 그리고 비교를 위한 sham구는 미크로좀에 DMSO만을 첨가하여 같은 조건으로 배양하였다. 그 결과, sham구의 CYP 함량은 배양 6시간이 지나서는 처음 수준의 91% 로 줄었으며, 0.1 mM과 0.2mM, 0.4mM 및 1.0 mM BNF 배양구는 배양 4 \~ 6시간 후에 최대값이 되었는데 각각 처음 보다 116% , 114% , 126% 및 132% 로 되었다. PB와의 배양에서는 0.1 mM과 0.4 mM 및 1.0 mM 배양구는 배양 4 \~ 6시간 후에 최대 값이 되었고 각각 처음의 112% , 114% 및 114% 로 되었으며, MC와의 배양에서도 0.1 mM과 0.4 mM 및 1.0 mM 배양구는 배양 4시간 후에 최대값이 되어 각각 116% , 130% 및 132% 로 증가하였다. 그리고 EROD 활성의 변화도 sham구는 배양 7시간 후에도 변화가 거의 없었으나, BNF의 0.1 mM과 0.4 mM 및 0.8 mM 배양구는 배양 7시간 후에 각각 처음의 120% , 123% 및 115% 로 되었고, PB의 0.1 mM과 1.0 mM 배양구는 배양 3시간 후에 최대로 되어 각각 121% 및 136% 로 증가하였으며, MC와의 배양에서는 0.1 mM과 0.4 mM 및 0.8 mM 배양구는 배양 5시간 후에 최대값이 되어 각각 204% , 198% 및 191% 로 증가하였다. 이처럼 BNF나 PB, MC는 명주조개에서도 미크로좀의 CYP 함량과 EROD 활성을 증가시키는 유도효과를 나타내었으며, 특히 MC에 의한 유도효과가 가장 강했고 BNF, PB의 순이었고, 대체로 약물의 농도가 높을수록 유도 효과도 큰 경향을 보였다. Induction of cytochrome P45O (CYP) and 7-etholqresorufin-O-deethylase (EROD) in the microsome exposed to 3-methylcholan-throne (MC), β -naphthoflavone (BNF) and phenobarbital-Na (PB) was investigated, Microsome was isolated from digestive gland of clam (Coelomactra antiquata) and then exposed to each chemical in concentration range of 0.1 to 1.0 mM for 7 hours. The CYP content and EROD activity in the microsome exposed to each chemical significantly increased compared to the control group. The overall CYP and EROD induction potency was in order of MC>BNF>PB. The induction response of EROD was two times higher than that of CYP level in the microsome exposed to MC, but the induction response of EROD was slightly higher than that of CYP level in BNF and PB exposure groups.33Nkciothe

Effects of Trialkyltin in vitro on the Microsomal Monooxygenase System of Digestive Gland in the Clam, Coelomactra antiquata

강원 북부연안에 많이 서식하는 명주조개 (C. antiquafa)를 대상으로 중장선의 미크로좀을 in vitro로 유기주석화합물 (TBTO, TBTC, TPTC)과 배양하여 약물대사효소계의 cytochrome P450 (CYP)과 7-ethoxyresorufin-O-deethylase (EROD)의 변화를 조사하였다. 그 결과, 이들 화합물은 모두 명주조개의 약물대사효소계를 짧은 시간 내에 저해한다는 것을 확인하였다. 즉,미크로좀을 0.4 mM 농도의 TBTC, TBTO 및 TPTC와 20분간 배양한 후의 CYP 함량은 첨가하기 전에 비해 각각 52% , 72% 및 40% 로 줄었는데, 이것으로 화합물의 종류에 따라서 저해 정도는 차이가 있었고 butyltin화합물보다는 phenyltin화합물의 저해 정도가 더 컸다. 그리고 EROD 활성의 경우도 0.4 mM의 TBTC, TBTO 및 TPTC와 20분 간 배양하였더니 각각 100% , 92% 및 85% 로 butyltin 화합물보다 는 phenyltin 화합물의 저해가 더 컸다. 한편, TBTC, TBTO 및 TPTC 모두는 패류 중장선의 CYP와 EROD 활성을 농도의존적으로 저해하였으며, 그리고 두 효소는 모두 오염물질에 노출된 패류의 좋은 생체지표 (bioindicator)로 활용할 수 있을 것이라 여겨진다. This study was carried out to measure the in vitro interaction of trialkyltin with the microsomal monooxygenase (MFO) system of the clam, Coelomactra antiquata. Cytochrome P450 (CYP) level and 7-ethonvesorufin-O-deethylase (EROD) activity were invetigated in the microsome isolated from digestive gland of the clam (C. antiquata) exposed to tributyltin chloride (TBTC), bis-tributyltinoxide (TBTO) and triphenyltin chloride (TPTC). The specific contents of CYP in clam microsome exposedto 0.4 mM TBTC, TBTO and TPTC for 20 minutes were decreased 52, 72 and 40% , respectively, compared to control group. The EROD activities also were inhibited by exposure to TBTO ( 92% ) and TPTC ( 85% ) except for TBTC, The level of CYP and the EROD actintles were decreased according to the OTC exposure concentrations. The toxic effects on the level of CYP and the EROD activities were in order of TPTC>TBTC>TBTO in this study. The measurement of CYP level and EROD activity could be applied as a biomarker for environmental study.33Nkciothe

In vitro Induction of Hepatic Cytochrome P450 (CYP) with Exposure to b-naphthoflavone in Marine Fishes

해산 어류가 cytochrome P450(CYP)유도제로 알려진 β-naphthoflavone(BNF)에 의해 어떤 반응을 하는지 살펴보기 위하여, 양식 어류로는 조피볼락(Sebastes schlegeli), 넙치(Paralichthys olivaceus), 참돔(Pagrus major)을 그리고 자연산 어류로는 숭어(Mugil cephalus)와 쥐치(Stephanolepis cirrhifer)를 대상으로 조사하였다. 숭어와 쥐치, 참돔의 미크로좀은 DMSO(dimethyl-sulfoxide)로 녹인 BNF를 10mM이 되도록 첨가하였고, 조피볼락과 넙치의 경우에는 5mM이 되도록 첨가하여 30℃의 수조에서 9시간까지 배양하면서 CYP함량의 경시적인 변화를 조사하였고, 각 어류는 2%농도로 DMSO만을 첨가하여 같은 조건으로 배양한 대조구에 대한 상대비(%)로 나타내었다. 그 결과, 대부분의 어류에서는 배양 3～5시간에 최대값을 보이고 이후 줄어들었으나 참돔은 9시간까지도 계속 증가하는 경향을 보였다. 한편, BNF에 의한 CYP 유도 정도는 양식산 어류가 자연산 어류보다 커서 차이를 보였다. 즉, 양식산 어류인 넙치, 조피볼락 및 참돔의 CYP 함량은 BNF의 노출농도를 감안하지 않더라도 각 어류의 처음 수준에 비해 207～422%까지 늘었던 것에 비해 자연산 어류인 숭어와 쥐치에서는 206～207% 증가하는데 불과하였다. 이처럼 양식산과 자연산 어류에서 CYP 유도제에 의한 CYP유도 정도의 차이는 서식환경의 차이에 따라 생기는 것이라 여겨진다. Cytochrome P450 (CYP) induction was determined in microsomes of three aquacultured fish species (Sebastes schlegeli, Paralichthys olivaceus and Pagrus major) and two wild fish species (Mugil cephalus and Stephanolepis cirrhifer) in vitro exposed to β-naphthoflavone (BNF). The microsomes of five fish were exposed to BNF (5 mM or 10 mM) in dimethylsulfoxide at 30℃ for 9 hr. The CYP contents in most fish increased according to exposure duration for 3 or 5 hour, and then decreased, while steady increase of CYP was observed in P. mqjor for 9 hour. The induction of CYP contents in aqua-cultured fish species (207 - 422%) were higher than those in wild fish species (206 -207%).33Nkciothe

Determination of Organic Contaminants in Sediments Using Trace Analysis and Biological Screening Assay

Gwangyang Bay located in the southern coast of Korea is a semi-enclosed embayment including a big steel mill plant and petroleum refinery plant complex around the bay. Several biomarkers and bioassays were accompanying with chemical analysis to monitor extent of contamination and effects of organic pollutants for the basic study to prepare a national action plan for protection of the marine environment from the land-based organic pollutants. Organic pollutants (PCBs, PAHs, organochlorine pesticides, organotins, alkylphenols, and phthalate esters) were analyzed in the surface sediment from the bay. In addition, bioassays, including four cytotoxicity tests (cell viability, esterase activity, double strand DNA content and cell membrane micro-viscosity) and E-screen assay with yeast transformants, were applied to sediment extract to evaluate toxicity and estrogenicity of sediments. Enzyme activity (EROD, MROD and GST) of feral fish collected from the polluted site of the bay and a reference site was measured as exposure biomarker for organic pollutants. Imposex in marine snail, were also applied to assess specific effect of organotin, especially tributyltin in the bay.Although the causality of effects on marine organisms were not clearly revealed in all the field studies, the measurable amount of certain effects in several bioassays indicate that organic toxicants in the coastal environment of Korea probably cause adverse effects on the marine organisms. However, more finely designed toxicity identification and evaluation (TIE) approach is required to confirm major stressors and to manage marine pollutions in the study area.1

Effects of Tributyltin in vitro on Hepatic Monooxygenase System in Marine Fishes

본 연구는 내분비교란물질로 알려진 TBTC가 해양생물에게 미치는 영향을 조사하는 연구의 일환으로, 이 화합물에 노출시킨 어류의 간장 MFO 효소계의 반응을 in vitro로 조사하였다. 대상 어류는 개복치(Mola mola), 숭어(Mugil cephalus), 강도다리(Ptatichthys stellatus), 청어(Clupeu pallasii), 붕장어(Astroconger myriaster), 조피볼락 (Sebastes schlegeli), 참돔(Pugrus major), 넙치(Paralichthys olivaceus)이며, 이들의 간장 미크로좀을 2mM의 TBTC와 in vitro (3 0∘C , 20분)로 배양하고 cytoch-rome P45O (CYP), cytochrome b5, NAD(P)H -cytochro-me c 환원효소를 비롯한 탈알킬화 효소들(EROD, PROD, MROD, ECOD)의 변화를 조사하였다. TBTC는 환원효소의 측정 시에는 DMSO에, 그리고 그 밖의 효소 측정시에는 메탄올에 녹여 2% 농도로. 첨가하였다. 어류의 간장 CYP 함량은 TBTC와 배양 후 대부분(6 어류/8어류)에서 10% 이하로 크게 저해되었으나, cyto-chrome b5함량은 변함이 없었다. 하지만 NAD(P)H 의존성 환원효소의 반응은 어류에 따라 달랐고, CYP 외에 두 환원효소도 모두 영향을 받아 저해되는 타입 1(개복치, 조피볼락, 청어, 강도다리, 참돔), CYP 외에 NADH 의존성 환원효소만 저해되는 타입 2(붕장어, 숭어)및 CYP 저해되지만 두 환원효소는 영향을 받지 않거나 오히려 유도되는 타입 3 (넙치)으로 구분되었다. 그리고 대부분(7/8)의 어류에서는 NADH 의존성 환원효소가 NADPH 의존성 환원효소에 비해 더욱 저해되는 경향을 보였다. TBTC는 어류의 탈알킬화 효소에도 영향을 미쳤고, 어류별 EROD활성의 저해는 개복치, 참돔, 붕장어, 조피볼락(잔존율 1~7%)>숭어, 청어 (14~30%)>넙치, 강도다리(56~65%)의 순이었으며, ECOD 활성의 저해도 개복치, 참돔, 붕장어 (36~38%)>조피볼락(63%))숭어, 청어, 넙치, 강도다리(90%)의 순으로 비슷한 경향이었다. 한편, 넙치와 강도다리에서는 PROD가 MROD보다 더욱 심하게 저해되었다. 이처럼 어류 간장의 약물대사 효소계는 TBTC에 의한 저해 정도가 어류에 따라 심한 차이를 보였다. Effects of tributyltin chloride (TBTC) in vitro on mixed function oxygenase (MFO) system on liver microsome of eight marine fish species were investigated. To determine the effects on MFO system, cytochrome P45O (CYP) and cytochrome b5 con-tents, activities of two reductases (NADH-cytochrome b5 reductase and NADPH-cy-tochrome P450 reductase) and four dealkylation enzymes (EROD, PROD, MROD and ECOD) were measured in fish microsoms exposed to TBTC for 20 min. The WP content was reduced to 10% of the control group in 6 out of 8 species exposed to TBTC, whereas there was no significant change in the cytochrome bs content. the response of NAD(P)H dependant reductases depended on fish species. The dealkylation enzyme activities in microsome were also apparently inhibited by TBTC. The degree of inhibition was different among fish species and four enzymes. The EROD activities in eight species were decreased to the range of 1∼65% of control group.33Nkciothe

Induction of vitellogenin in the rockfish exposuredto 2,2',4,4&quot5,5'-hexachlorobiphenyl(153): comparison of gene expression and protein level

The level of r VTG mRNA expression increased only in female rockfish2

Toxicity of Organotin Compounds on the Survival of Rotifer (Brachionus plicatilis)

유기주석화합물은 농업과 산업분야에서 많이 쓰이고 있으며, 이들은 수계환경으로 유입되어 물이나 저질 중의 세균들에 의해 분해되어 최종적으로는 무기주석으로 된다. Trialkyltin 화합물이 해양생물에 미치는 영향에 관해서는 많은 연구가 있지만, 해양생태계에서 기초적인 생산을 담당하는 플랑크톤류에 미치는 영향에 관해서는 잘 알려져 있지 않다. 본 연구에서 TBT와 TPT의 분해물인 DBT, MBT, DPT, MPT를 비롯하여 trimethyltin(TMT)과 그 분해물인 dimethyltin(DMT)이 기수산 rotifer의 생존에 미치는 독성(95hr-LC_(50))을 조사ㆍ비교하였다. 그 결과, DPT(13.8ppb)가 가장 강했으며 TMT(42.9), DBT(80.6), MPT(262.2), MBT와 DMT(>1,000)의 순으로 나타났다. 즉, trialkyltin과 dialkyltin에서는 폐닐주석, 부칠주석, 메칠주석의 순으로 독성이 약해지며, monoalkyltin에서도 폐닐주석화합물이 메칠주석화합물에 비해 독성이 강하다는 것을 확인할 수가 있었다. 이렇듯 독성이 서로 다른 것은 유기주석화합물이 소수성(lipophilicity)과도 높은 상관이 있었다.33Nkciothe