• Journal of fish pathology
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• v.26 no.3
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• pp.219-229
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• 2013

Experiments were carried out to investigate the growth and hematological parameters of rock bream Oplegnathus fasciatus exposed to several different concentrations (0, 1, 2, 4, $8{\mu}g/L$) of tributyltin (TBT) for 4 weeks. Growth rate of the fish exposed to $8{\mu}g/L$ of TBT concentration was significantly lower than that of control group fish. The major hematological findings were significant decreases in the red blood cell count, hematocrit value and hemogobin concentration in the fish exposed to $8{\mu}g/L$. Although serum glucose concentration was significantly reduced compared to the control group, total protein concentration was found to be significantly increased over the control group. Exposure to $8{\mu}g/L$ of TBT concentration resulted in significant increase in the enzyme activities, such as glutamic oxalate transminase and glutamic pyruvate transminase in the fish. The present findings suggest that exposure to $8{\mu}g/L$ concentration of TBT can cause significant changes in growth and hematological parameters of rock bream Oplegnathus fasciatuso.

• Animal cells and systems
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• v.11 no.1
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• pp.1-8
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• 2007

Recently, we have shown that some endocrine disruptors, heavy metals, organotins and azoles suppressed steroidogenic enzymes such as P450 side-chain cleavage enzyme (P450scc) and aromatase in bullfrog ovarian follicles. In the present study, by using an amphibian ovarian follicle culture system, we examined the effects of these endocrine disruptors on maturation and ovulation of oocytes from Rana dybowskii in vitro. Ovarian fragments or isolated follicles were cultured for 24 h in a medium containing frog pituitary homogenate (FPH) or progesterone ($P_{4}$) with or without endocrine disruptors, and oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation were examined. Among the organotins, tributyltin (TBT) strongly inhibited both FPH-and $P_{4}-induced$ oocyte maturation ($ED_{50}$:0.6 and 0.7 ${\mu}M$, respectively); however, tetrabutyltin (TTBT) and dibutyltin (DBT) showed only partial suppression, while monobutyltin (MBT) showed no inhibitory effect. All of the organotins suppressed $P_{4}-induced$ oocyte ovulation very effectively at a low concentration, and TBT and DBT exerted an inhibitory effect on FPH-induced ovulation. Among the heavy metals, mercury (Hg), cadmium (Cd) and cobalt (Co) were very effective in inhibiting FPH-induced oocyte maturation and ovulation, while lead (Pb), arsenite (As) and zinc (Zn) were less effective. However, all of the heavy metals suppressed FPH-induced oocyte ovulation at a high dose ($100{\mu}M$). Among the azoles, itraconazole (ICZ), ketoconazole (KCZ) and clotrimazole (CTZ) effectively inhibited FPH-induced oocyte maturation and ovulation, while econazole (ECZ), miconazole (MCZ) and fluconazole (FCZ) were considerably less effective. These results demonstrated that the abovementioned endocrine disruptors exhibited differential effects on oocyte maturation and ovulation in amphibian follicles and that the frog ovarian culture system could be used as an effective experimental tool to screen and evaluate the toxicity of various endocrine disruptors in vitro.

• Journal of the Korean Society for Marine Environment & Energy
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• v.9 no.1
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• pp.21-28
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• 2006

Since the usage of tributyltin(TBT) has been banned, many chemicals including Sea-Nine 211, Cu-pyrithione, and Zn-pyrithione were developed to use as antifouling agents for ships and coastal structures. However, the toxicity of these antifouling chemicals have not been systematically evaluated in ecotoxicological and biological studies. In this study, we investigated the effect of four antifouling substances on survival of estuarine rockfish, Sebastes schlegeli and amphipod, Monocorophium acherusicum. Survival of S. schlegeli and M. acherusicum during the 96-h exposure period were used to estimate the median lethal concentrations(LC50s) of test chemicals for each test species. Among antifouling agents, Cu-pyrithione($56{\mu}g{\cdot}1^{-1}$;96-h LC50) was most toxic to S. schlegeli, followed by $TBT(73{\mu}g{\cdot}1^{-1}),\;Sea-Nine(184{\mu}g{\cdot}1^{-1})\;and\;Zn-pyrithione(l707{\mu}g{\cdot}1^{-1})$, while TBT($26{\mu}g{\cdot}1^{-1}$) was most toxic to M. acherusicum followed by Sea-Nine($49{\mu}g{\cdot}1^{-1}$), Cu-pyrithione($119{\mu}g{\cdot}1^{-1}$) and Zn-pyrithione($334{\mu}g{\cdot}1^{-1}$). Effect concentrations of the antifouling chemicals estimated in this study can be used when assessing the potential risks of these substances, of which usage is increasing in the coastal environment.

• Korean Journal of Environmental Biology
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• v.25 no.4
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• pp.313-318
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• 2007

Monitoring for the quality of coastal water is necessary to improve its biological resources and to maintain healthy environments. We measured the concentrations of phenols, nine polychlorinated biphenyls (PCBs), tributyltin (TBT) in surface water at 5 stations and the concentrations of mercury (Hg), chromium (Cr (VI)), cadmium (Cd), lead (Pb) in surface water of 10 stations in Tongyeong marine ranching ground of Korea in August, 2003. The concentrations of analytes were determined as follows: phenols: 1.6$\sim$2.8 ppb, PCBs: not detected (ND), Hg: <0.1 ppb, Cr (VI): 0.01$\sim$5.32 ppb, Cd: ND$\sim$0.41 ppb and Pb: 0.43$\sim$2.60 ppb. These concentrations satisfied the standards of human health protection in coastal water by Ministry of Environment, Republic of Korea as follows: phenols; 5 ppb, PCBs: 0.5 ppb, Hg: 0.5 ppb, Cr (VI): 50 ppb, Cd: 10 ppb and Pb: 50 ppb. TBT was not listed in the standards. The concentrations of Sn in TBT analyzed in sea water of Tongyeong marine ranching ground were in the range of ND$\sim$0.0273 ppb which are similar with the values of 8$\sim$35 ng Sn $L^{-1}$ (0.008$\sim$0.035 ppb) in Chinhae Bay studied by Shim et al.. Therefore, the quality of sea water in Tongyeong marine ranching ground was safe enough to protect human health.

• Journal of fish pathology
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• v.21 no.1
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• pp.45-56
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• 2008

This study was conducted to find out biological responses of bivalves exposed to organotin compound.The results of the study confirmed that tribultyltin chloride (TBTCl) induce reduction of survival rate andburrowing activity, and histopathological feature in the foot structure of the equilateral venus, Gomphinaveneriformis. The experimental period was 36 weeks. The experimental groups consisted of a control and 3TBTCl exposure groups (0.4, 0.6, 0.8 ym TBTCl L'). The survival rate and burrowing activity were record-ed daily. For histological analysis, foot tissues were fixed in Bouin' s fluid and then stained H-E stain, AB-PAS (PH 2.5) reaction and Masson's trichrome stain after having serially sectioned the tissue by paraffinmethod at thickness of 4-6 ym. The survival rate was not significantly different between the control andexposure groups for 20 weeks, but in 0.8 Um TBTCl L', it was on the decreased ever since the exposure. Theburrowing activity was not significantly different in the exposure group compared to the control up to 12weeks, but in 0.6 and 0.8 ym TBTCl L', it measured the lowest level after 20 weeks. The foot is composedof the epidermal layer, connective tissue, and muscular layer. The epidermal layer is composed of simplecolumnar, cuboidal epithelia and mucous cells. The cilia were well developed on the apical surface ofepithelium, Circular, longitudinal and transverse muscle bundle were well developed in the muscular layer.The majority mucous cells showed blue color (542c) when it subjected to AB-PAS (PH 2.5) reaction. Nohistopathological alterations in the foot were observed up to 12 weeks. After 20 weeks of exposure to 0.8 (anTBTCl L'', the foot samples of exposed G. veneriformis showed disappearance of cilia and striated borderpartially and extension of hemolymph sinus. The mucous cell increased in the marginal of foot. At 28-weekof exposure to 0.4 ym TBTCl L', it observed weekly acid (564c), neutral (264c) and mixed mucous cell. At36-week of exposure to 0.6 ym TBTCl L', it showed fragmentation of the muscle and collagen fiber bundle,and also diappearance of cilia on epithelia and edema of epithelium in 0.8 ym TBTCl L''.

• Development and Reproduction
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• v.13 no.3
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• pp.163-172
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• 2009

Some organotin compounds such as butyltins and phenyltins are known to induce impo-sex in various marine animals and are considered to be endocrine disruptors. In this study, the effect of organotins on follicular steroidogenesis in amphibians was examined using ovarian follicles of Rana dybowskii and Rana catesbeiana. Isolated follicles were cultured for 6 or 18 h in the presence and absence of frog pituitary homogenate (FPH) or various steroid precursors, and the levels of product steroids in the culture media oassay. Among the butyltin compounds, tributyltin (TBT) strongly and dose-dependently inhibited the FPH-induced synthesis of pregnenolone ($P_5$) and progesterone ($P_4$) by the follicles. TBT also strongly suppressed the conversion of cholesterol to $P_5$ and partially suppressed the conversion of $P_5$ to $P_4$. A high concentration of dibutyltin (DBT) also inhibited steroidogenesis by the follicles while monobutyltin and tetrabutyltin had negligible effects. The toxic effect of TBT or DBT was irreversible and a short time of exposure (30 min) was enough to suppress steroidogenesis. All the phenyltin compounds significantly inhibited FPH-induced $P_5$ synthesis by the follicles. The effective dose of 50% inhibition by diphenyltin was $0.04\;{\mu}M$ and those of monophenyltin and triphenyltin were $0.24\;{\mu}M$ and $0.3\;{\mu}M$, respectively. However, none of the phenyltin compounds significantly suppressed the conversion of $P_4$ to $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) (by $17{\alpha}$-hydroxylase), $17{\alpha}$-OHP to androstenedione (AD) (by $C_{17-20}$ lyase), or AD to testosterone by the follicles. Taken together, the data show that among the steroidogenic enzymes, P450scc in the follicles is the most sensitive to organotin compounds and that an amphibian follicle culture system can be a useful screening model for endocrine disruptors.

• Korean Journal of Environmental Biology
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• v.26 no.2
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• pp.102-108
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• 2008

Many alternative biocidal additives were applied to antifouling paint to replace TBT, and Sea-Nine 211 is one of alternating organic booster compounds used in antifouling paint. In this study, extent of Sea-Nine 211 toxicity on marine benthic bivalve is evaluated. Sea-Nine 211 was injected to surf clam, Pseudocardium sachalinensis, that inhabitate northern part of Gangwon Province, Korea. Survival rate of the clam and xenobiotics metabolizing enzyme activities in digestive gland were measured during 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$^{-1}$ body weight), while the clam exposed to 1, 2 and 5 mg kg$^{-1}$ TBT chloride (TBTC) demonstrated 70, 30 and 0% survival rate, respectively. The Sea-Nine 211 exposure group showed a tendency of cytochrome P450 (CYP) induction according to the exposure duration, on the other hand, CYP content was decreased in the TBT exposure group. NADPH cytochrome P450 reductase activity slightly increase according to the exposure duration in the Sea-Nine 211 exposure group, while TBTC inhibit its activity as CYP content. Moreover, there was no significant change of NADH cytochrome b5 reductate activity in the clam epxosed 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. All the results indicate that Sea-Nine 211 demonstrated a tendency to induce CYP level, while TBTC inhibits the CYP level, NADPH cytochrome P450 reductase and NADH cytochrome b5 reductase activities.

• Journal of the Korean Society for Marine Environment & Energy
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• v.12 no.2
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• pp.68-74
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• 2009

Polycyclic aromatic hydrocarbons (PAHs), one of ubiquitous organic pollutants in marine environments, are major toxic components of petroleum and are produced during the incomplete combustion of organic materials. As shipyards are located inside of natural or artificial semi-enclosed bay, even a relatively weak environmental disturbance by ship-building activity can cause severe damage to marine ecosystem in the bay. Many studies of pollution in shipyard area have been focused on the antifouling agent, like tributyltin. This study aimed to investigate the effect of ship-building activity on PAH contamination. Total PAHs concentration was higher nearby and inside shipyard area than outside, implying that shipyard could be one of major source area of PAH contamination to pose harmful effects to surrounding environments. Through PAH profile and source recognition index, the source of PAHs inputs in this area was estimated to originate from both petrogenic and pyrogenic origin. PAH levels showed a significant correlation with total butyltins, indicating that ship-building activity influenced PAH concentration and distribution. Vertical distribution of PAHs historically confirmed the correlation between shipbuilding activity and PAHs contamination.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.378-384
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• 2012

In aquatic invertebrates, particularly marine gastropods, organotin compounds induce irreversible sexual abnormality in females, which is termed imposex, at very low concentrations. Organotin compounds are agonists for nuclear receptors such as RXRs and $PPAR{\gamma}$. However, the imposex phenomenon has not been reported to act as an antagonist on estrogen receptors in other species, including vertebrates and invertebrates. In order to gain insights into the antagonistic activity of organotin compounds on estrogen receptors (ERs), we examined the inhibitive effect of these compounds on estradiol-dependent ${\beta}$-galactosidase activity using the yeast two-hybrid detection system consisting of a combination of the human estrogen receptor ($hER{\beta}$) ligand-binding domain and the co-activator steroid receptor co-activator-1 (SRC1). Tributyltin-hydroxide (TBT-OH) and triphenyltin-chlorine (TPT-Cl) exhibited an inhibitive effect on $E_2$-dependent transcriptional activity, similar to antagonistic chemicals such as 4-hydroxytamoxifen (OHT) or ICI 182,780, at a very low concentration of $10^{-14}$ M TBT or $10^{-10}$ M TPT, respectively. The yeast growth and transcriptional activity with transcriptional factor GAL4 did not exhibit any effect at the tested concentration of TBT or TPT. Moreover, the yeast two-hybrid system using the interaction between p53 and the T antigen of SV40 large did not describe any effect at the tested concentration of OHT or ICI 182,780. However, the interaction between p53 and T antigen was inhibited at a TBT or TPT concentration of $10^{-9}$ M, respectively. These results indicate that TBT and TPT act as inhibitors of ER-dependent reporter gene transcriptional activation and of the interaction between $hER{\beta}$ LBD and the co-activator SRC1 in the yeast two-hybrid system. Consequently, our data could partly explain the occurrence of organotin compound-induced imposex on the endocrine system of mammals, including humans.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.185-190
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• 2002

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.