Adsorption of tributyltin by tributyltin resistant marine Pseudoalteromonas sp. cells

The isolate, Pesudoalteromonas sp. TBT1, could grow to overcome the toxicity of tributyltin chloride (TBTCl) up to 30 μM in the absence of Cl⁻ in the medium until the cells reached an exponential phase of growth. The viability, however, was reduced after the cells reached a stationary phase. The degradation products, such as dibutyltin (DBT) and monobutyltin (MBT), were not detected in the growth medium, indicating that the isolate has no ability to degrade TBT into less toxic DBT and MBT. Up to about 10^7.5 TBT molecules were adsorbed by a single cell. The observation of morphological changes with an electron microscope showed that the cell surface became wrinkled after exposure to the lethal concentration of 10 mM TBTCl. These results indicate that the resistance of the isolate toward the toxicity of TBTCl is not related to the unique cell surface, which seems to play an important role in preventing the diffusion of TBTCl into the cytoplasm.     

Adsorption of tributyltin by tributyltin resistant marine Pseudoalteromonas sp. cells

The isolate, Pesudoalteromonas sp. TBT1, could grow to overcome the toxicity of tributyltin chloride (TBTCl) up to 30 μM in the absence of Cl⁻ in the medium until the cells reached an exponential phase of growth. The viability, however, was reduced after the cells reached a stationary phase. The degradation products, such as dibutyltin (DBT) and monobutyltin (MBT), were not detected in the growth medium, indicating that the isolate has no ability to degrade TBT into less toxic DBT and MBT. Up to about 10^7.5 TBT molecules were adsorbed by a single cell. The observation of morphological changes with an electron microscope showed that the cell surface became wrinkled after exposure to the lethal concentration of 10 mM TBTCl. These results indicate that the resistance of the isolate toward the toxicity of TBTCl is not related to the unique cell surface, which seems to play an important role in preventing the diffusion of TBTCl into the cytoplasm.     

Differences in the acute toxicities of tributyltin between the Caprellidea and the Gammaridea (Crustacea: Amphipoda)

Tests for the acute toxicity of tributyltin (TBT) were conducted on amphipod crustaceans collected from Otsuchi Bay, Japan. Five species of caprellids and three species of gammarids, which belong to a closely related ecological niche, were used for the exposure experiments at seven test concentrations (0, 0.001, 0.01, 0.1, 1, 10 and 100 microg TBTCl/l) for 48 h at 20 degrees C. The 48-h LC50 values of the caprellids were 1.2-6.6 microg TBTCl/l, and these were significantly lower than those of the gammarids (17.8-23.1 microg TBTCl/l). This suggests that caprellids are more sensitive to TBT than gammarids. Furthermore, the proportions of TBT and its derivatives, dibutyltin (DBT) and monobutyltin (MBT), were measured in the amphipods collected from Otsuchi Bay. In the caprellids, TBT was the predominant compound, accounting for 72% of the total butyltin which reflected the butyltin ratio in seawater, while in the gammarids, TBT's breakdown products (DBT and MBT) predominated, accounting for 75% of the total butyltin. This difference suggests that caprellids may have lower metabolic capacity to degrade TBT than gammarids. Therefore, the difference in sensitivity to TBT among the amphipods is thought to be related to the species-specific capacity to metabolize TBT.     

Removal of tributyltin (TBT) by live and dead microalgal cells

The removal of tributyltin (TBT) from artificial wastewater by dead and live cells of four microalgal species, Chlorella miniata, C. sorokiniana, Scenedesmus dimorphus and S. platydiscus was investigated. Dead cells were generally more efficient in removing TBT during three days exposure. More than 90% and 85% TBT were removed by dead cells of Scenedesmus and Chlorella, respectively. Although the removal percentages of live microalgae were smaller, TBT was continuously absorbed and degraded, and TBT removal percentages similar to respective dead cells were achieved at the end of 14 days treatment. Degradation products, DBT and MBT, were recorded mainly inside the cells, and intracellular MBT concentrations were significantly higher than DBT. In terms of TBT removal, Scenedesmus cells were more effective than Chlorella, probably due to larger cell sizes and biomass. However, TBT specific uptake and degradation by Chlorella was higher than in Scenedesmus. The highest specific TBT uptake values, as well as the highest degradation, were recorded in C. miniata, a Hong Kong isolate.     

Chronic effects of tributyltin on the caprellid amphipod Caprella danilevskii

In order to examine the biological effects by TBT exposure, experiments involving the exposure of five levels of TBT concentrations (0, 10, 100, 1000 and 10,000 ngl(-1)) were conducted on the caprellid amphipod Caprella danilevskii, over a generation after hatching. Marked delays in growth and molting during the early developmental stage and maturation were found in both 100 and 1000 ng TBTCl l(-1) concentrations in spite of the sex. All specimens died in 10,000 ng TBTCl l(-1) within 4 days after hatching. Inhibition of maturation and reproduction such as delaying in the achievement of maturity and a decrease in the number of juveniles hatched was apparent in 10 and 100 ng TBTCl l(-1) concentrations. Furthermore, brood loss, and failure in egg formation and hatching were observed as the TBT concentration became higher. No significant changes in sex ratio were seen at any of the TBT exposure levels during hatching and maturation in the present study. This phenomenon was different from a previous study in which the proportion of females was increased with an increase in TBT concentrations in the embryonic exposure experiment. This suggests that sex disturbance is induced during the embryonic stage of the species. A drastic decrease in survival rate was observed at 10 ng TBTCl l(-1) (25%) which corresponds to the mean level in coastal waters. Therefore, the present encountering ambient TBT concentration may influence populations of C. danilevskii in the coastal environment.     

Status of butyltin pollution along the coasts of western Japan in 2001, 11 years after partial restrictions on the usage of tributyltin

Restrictions on the use of tributyltin (TBT) in aquaculture and on boats in coastal regions, except for ocean-going vessels, have been in place in Japan since 1990 due to the strong toxic effects of TBT on marine organisms. However, TBT pollution along the Japanese coasts has been reported after this legislation was enacted. In order to elucidate the present status of contamination by butyltin (BT) compounds, we measured the levels of BTs [monobutyltin (MBT), dibutyltin (DBT) and TBT] in seawater and Caprella spp. samples obtained from the western part of Seto Inland Sea, Uwa Sea and Uranouchi Bay in western Japan during March to September, 2001. Butyltins were detected in more than 90% of the seawater samples (n = 59), with average concentrations of 8.2 ± 9.2 (SD) ng MBT L⁻¹, 3.3 ± 3.0 ng DBT L⁻¹ and 9.0 ± 7.0 ng TBT L⁻¹. Among 41 stations situated on coastal lines, a sufficient number of Caprella organisms for chemical analysis could be collected from only 16 stations. The butyltin concentrations in seawater and Caprella samples from Uwa Sea and Uranouchi Bay, in which the dominant industry in both waters is aquaculture, showed significantly higher than or no significant differences from those samples from the western part of Seto Inland Sea, a major heavy-industry area in Japan. As the TBT concentration in seawater increased, the number of Caprella organisms collected decreased. The mean TBT concentration among the seawater samples was above the estimated lowest observable effect concentration (LOEC) that reduces the survival rate of Caprella danilevskii. Thus, the present study indicates that TBT is still a potential ecological hazard to the survival of marine invertebrates inhabiting coasts along western Japan, even 11 years after the partial ban on TBT usage was implemented.     

Assessment of organotin (butyltin species) contamination in marine biota from the Eastern Aegean Sea, Turkey

The marine environment continues to be adversely affected by tributyltin (TBT) release from maritime traffic. Therefore the concentrations of TBT, dibutyltin (DBT) and monobutyltin (MBT) were measured in barnacles, mussels and fish along the Eastern Aegean coastline. The average concentrations of TBT ng Sn g⁻¹ were found to be 235 in fish, 116 in mussels and 635 in barnacles. The highest concentrations of TBT, DBT and MBT were observed in the barnacles which had been sampled in marinas and harbors. All mussels sampled showed values of TBT + DBT, which were below the “tolerable average residue level (TARL)” as currently accepted. This indicates a lack of risk to the consumer. However, 7 out of the 15 fish sampled displayed TBT + DBT levels above the TARL, which indicates that a fish consumer group may be at risk. Barnacles have high potential as biomonitors for the presence of organotin in the Aegean Sea.     

Integrative assessment of organotin contamination in a southern European estuarine system (Ria de Aveiro, NW Portugal): tracking temporal trends in order to evaluate the effectiveness of the EU ban

Organotin (OT) compounds have been used as biocide agents in antifouling paints since the mid 1960s and are now ubiquitous in the marine environment. Due to their high toxicity to non-target species they were banned from antifouling paints in the European Union in 2003 (2002/62/EC directive). The aim of the present work is to assess any obvious decline of the OT environmental levels at Ria de Aveiro (NW Portugal) after the ban. The organotins - monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), monophenyltin (MPT), diphenyltin (DPT), triphenyltin (TPT), monoctyltin (MOT), dioctyltin (DOT) and trioctyltin (TOT) - were quantified in the gastropod Nassarius reticulatus, in the mussel Mytilus galloprovincialis and in sediments. Imposex (imposition of male characters on females of gonochorist gastropods) in N. reticulatus was additionally used as a biomarker of TBT pollution. Time comparisons show a slight decrease of imposex between 2003 and 2005 probably as a consequence of the EU ban, though in some cases this trend seems to have started earlier since 2000. The fraction of TBT relatively to its metabolites has been decreasing over the last years but still remains high suggesting that there are still recent inputs of this compound into the study area.     

Testes specific accumulation of tributyltin in turbot Scophthalmus maximus from the southern Baltic Sea

Concentrations of tributyltin (TBT) and its breakdown products, dibutyltin (DBT) and monobutyltin (MTB), were measured in tissues of male turbot Scophthalmus maximus from the Gulf of Gdańsk (GDA) and the Pomeranian Bay (POM) in the southern Baltic Sea. The fish (n = 206) were collected during the spawning seasons of 2008-2009. All specimens were at the same maturity stage and in full breeding condition. The ripe testes of the turbot appeared to be a target tissue for TBT accumulation. We report that TBT is extensively eliminated at the specific whole-body level and that the males from TBT-contaminated location (GDA) reach this specific level at 23-24 cm in total length.     

Concentrations of organotin compounds in blue mussels from the wharves of Tokyo Bay

Organotin compounds (tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT), triphenyltin (TPT), diphenyltin (DPT), and monophenyltin (MPT)) were quantified in blue mussels, Mytilus edulis, in 1989 at 26 wharves along Tokyo Bay. Organotin compounds, other than DPT, were detected in all mussel samples (in case of DPT: 23 of 26). The concentration of DBT was highest among butyltin compounds and that of TPT was highest among phenyltin compounds. The results showed that relatively high concentrations of organotin compounds persist in blue mussels of Tokyo Bay, although the use of the paints containing these compounds has been limited for several years by regulations.     

Baseline of butyltin pollution in coastal sediments within the Basque Country (northern Spain), in 2007-2008

Tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT) were measured in surficial sediments at, the ports of Pasaia and Bilbao, together with other mid- and small-size harbours of the Basque Country (northern Spain), in 2007-2008. The highest values of the sum of the three measured butyltin species (3523-3640 ng g⁻¹, as Sn) were found at sampling stations near to shipyards located within the port of Pasaia. The highest value of TBT concentration (3143 ng g⁻¹, as Sn) was found at the marina of Getxo, in the port of Bilbao. The degree of TBT degradation varied greatly between sampling stations, being found to be generally higher in those sediments with higher values of redox potential and lower values of TBT concentration (normalized by organic matter content).     

Toxicity of tributyltin in the marine mollusc Mytilus edulis

Our previous studies have demonstrated that tributyltin (TBT) is genotoxic to the early life stages of marine mussels and worms. Here, the toxicity of TBT to adult organisms was determined using a suite of biomarkers designed to detect cytotoxic, immunotoxic and genotoxic effects. Exposure of adult mussels, Mytilus edulis, to environmentally realistic concentrations of TBTO for 7 days resulted in a statistically significant decrease in cell viability at concentrations of 0.5 μg/l and above. TBT had no effect on phagocytic activity or antioxidant capacity (FRAP assay). There was a statistically significant increase in DNA damage detected using the comet and micronucleus assays between the controls and 0.5, 1 and 5 μg/l of TBTO (P > 0.0005). Furthermore there was a strong correlation between DNA strand breaks (comet assay) and formation of micronuclei (P = 0.0005; R² = 61.5%). Possible mechanisms by which TBT could damage DNA either directly or indirectly are discussed including the possibility that TBT is genotoxic due to its ability to disrupt calcium homeostasis.     

Organotins (TBT and DBT) in water, sediments, and gastropods of the southern Venice lagoon (Italy)

The release of tributyltin (TBT) from maritime traffic represents one of the main problems of direct, diffuse, and continued contamination of the marine environment. In the present survey, the concentrations of TBT and dibultytin (DBT) in brackish waters, sediments, and the gastropods Nassarius nitidus were evaluated in order to estimate the contamination of the southern part of the Venice lagoon. TBT and DBT were determined by GC-MS/MS. Recent contamination of TBT was found in brackish waters near marinas, whereas the highest concentrations of TBT and DBT were observed in surface sediments at dockyards and harbours. High content of organotin in the gastropods sampled near the dockyards, harbours, and marinas showed a mobilisation from the sediments through the food web. The present study allowed assessment of whether, despite the ban on the use of TBT paints, waters, sediments, and biota were still being contaminated by organotin compounds in the southern Venice lagoon.     

Organotin levels in Nazaré canyon (west Iberian Margin, NE Atlantic) and adjacent coastal area

Organotin compounds (OTs) are ubiquitous in the marine environment and high concentrations (μg g(-1) range) in sediments from different coastal areas around the world have been reported. However, few reports have described the OTs contamination status in the offshore and deep sea environment. This work investigated organotin levels in Nazaré canyon for the first time. Levels of monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), diphenyltin (DPT), triphenyltin (TPT), dioctyltin (DOT) and trioctyltin (TOT) were quantified in sediment samples from the upper flanks of the canyon and from the adjacent coastal area. TBT levels detected in the canyon flanks are about two to three orders of magnitude lower than those found in the coastal area. Nevertheless, when quantifiable, TBT levels in the canyon samples were higher than the Environmental Assessment Criteria set for TBT in sediments by the OSPAR Commission indicating that at those locations negative ecological impacts are likely to occur.     

Effects of tributyltin (TBT) on the seagrass Ruppia maritima

The effects of tributyltin (TBT) on the seagrass Ruppia maritima were studied in two growth experiments. Plants were sampled at stations in Odense Fjord and Lunkebugten, Denmark, and replanted in reference sediment without TBT, reference sediment spiked with TBT, and in impacted sediment sampled in the highly TBT contaminated (7-57 microg kg (-1) dw) Odense Fjord. Plant performance was studied at weekly intervals for 3-4 weeks, by measuring net photosynthetic activity, respiration, relative growth rate (RGR) and number of leaves. Net photosynthetic activity in plants from spiked and impacted sediment was reduced by up to 60% relative to reference plants. Respiration both increased and decreased in response to TBT exposure, while RGR was generally lower in plants from contaminated sediments (reduced by 8-25%). The effects of spiked and impacted sediment differed between the experiments, which could be partly explained by the bioavailability of TBT in the two treatments, but also by adaptation of the plants from Odense Fjord to TBT. Measurements of enhanced TBT concentrations in the sediments in Odense Fjord suggest an impact of TBT on R. maritima is possible under in situ conditions.     

Assessment of tributyltin contamination in a shipyard area using a mussel transplantation approach

Mussels (Mytilus galloprovincialis) were transplanted to seven stations around a large shipyard for 126 days to evaluate tributyltin (TBT) contamination. Although the application of TBT-based paints to ships is totally banned in Korea, butyltin compounds were found to accumulate in mussels following transplantation. Concentrations of TBT and total butyltins in transplanted mussels near the shipyard were in the range of 40-350 ng Sn/g and 74-530 ng Sn/g on a dry weight basis, respectively. Obviously, low TBT concentrations (6.0-53 ng Sn/gdw) were determined in mussels at four stations outside the shipyard. A negative gradient of TBT concentrations and TBT portion to total butyltin concentrations were found in both the surface water and transplanted mussels according to distance from the shipyard. In addition, TBT concentrations in surface water and transplanted mussels showed significant correlation (r(2) = 0.71; p < 0.001). These results indicate that the shipyard still releases fresh TBT into surrounding waters even after TBT regulation in Korea, and mussel transplantation is useful in evaluating TBT contamination in shipyard area.     

Decade-long monitoring reveals a transient distortion of baseline butyltin bioaccumulation pattern in gastropods

Worldwide measures to restrict tributyltin (TBT) in antifouling paints have been legislated for decades, and were upgraded to a total ban on September 2008. With a view to test the response of coastal biota to changing pollution, since 1996 we have determined the concentration of TBT and derivatives di- and mono-butyltin (DBT and MBT) in NW Spain populations of two gastropods of contrasting biology, the rock-snail Nucella lapillus (n = 18) and the mud-snail Nassarius reticulatus (n = 24). TBT pollution in the study area has decreased consistently and considerably over time. In addition, the baseline butyltin (BT) bioaccumulation patterns showed a marked but transient distortion. These field observations are consistent with BT desorption from sediments, a natural phenomenon that is now to be expected in developing countries recently subject to the global TBT ban.     

Distribution of butyltins in the waters and sediments along the coast of India

Water and surface sediment samples were analyzed for butyltins (TBT, DBT, MBT) from various ports along the east and west coast of India. The total butyltin (TB) in water samples varied between ∼1.7 and 342 ng Sn l⁻¹, whereas for sediments it varied between below detection limit to 14861 ng Sn g⁻¹ dry weight of sediment. On an average Chennai port recorded the highest level of butyltins in the sediments while Paradip recorded the highest level of butylins in the waters. A fairly good relationship between the TB in the sediment and overlying water samples, as well as between organic carbon and TB, implicates the importance of adsorption/desorption process in controlling the levels of TBT in these port areas. In India the data on organotin pollution is very sparse; most of the port areas have been surveyed for butyltins for the first time during this study.     

Toxicity of tributyltin in the marine mollusc Mytilus edulis

Our previous studies have demonstrated that tributyltin (TBT) is genotoxic to the early life stages of marine mussels and worms. Here, the toxicity of TBT to adult organisms was determined using a suite of biomarkers designed to detect cytotoxic, immunotoxic and genotoxic effects. Exposure of adult mussels, Mytilus edulis, to environmentally realistic concentrations of TBTO for 7 days resulted in a statistically significant decrease in cell viability at concentrations of 0.5 μg/l and above. TBT had no effect on phagocytic activity or antioxidant capacity (FRAP assay). There was a statistically significant increase in DNA damage detected using the comet and micronucleus assays between the controls and 0.5, 1 and 5 μg/l of TBTO (P > 0.0005). Furthermore there was a strong correlation between DNA strand breaks (comet assay) and formation of micronuclei (P = 0.0005; R² = 61.5%). Possible mechanisms by which TBT could damage DNA either directly or indirectly are discussed including the possibility that TBT is genotoxic due to its ability to disrupt calcium homeostasis.     

Imposex in Morula Granulata as Bioindicator of Tributyltin (TBT) Contamination in the Dampier Archipelago, Western Australia

This study was undertaken to examine the use of imposex in the tropical neogastropod whelk Morula granulata as bioindicator of tributyltin (TBT) contamination. Samples were collected from sites throughout the Dampier Archipelago; both impact sites close to intensive shipping activity and remote control sites. TBT in this area originates mainly from commercial vessels using the Port of Dampier that are >25 m in length and exempt from legislation controlling the use of TBT in antifouling paints. A field survey was undertaken in July 1997; 100 M. granulata were taken at each of 18 sites with varying vessel activity, and examined for imposex. The level of TBT contamination in the Archipelago was low; ranging from <0.3 to 25 ng Sn l⁻¹ in water,<0.3 to 80 ng Sn g−¹ in oyster tissue (Saccostrea cuccullata – prey of M. granulata), and <0.3 to 33 ng Sn g⁻¹ in whelks. Percentage of imposex in M. granulata ranged from 0% to 57%, and was correlated with distance to vessel activity (r=0.646, p<0.05). The only significant relationship of imposex with butyltin concentrations was with TBT in oysters (r=0.826, p<0.05), and dibutyltin (DBT) in whelks (r=0.783, p<0.05). It is postulated that this is because of interspecies variation in ability to metabolize TBT to DBT and MBT (monobutyltin), as well as induction of imposex in earlier life stages, so that imposex in adults could be a reflection of prior contamination. M. granulata is a suitable bioindicator of TBT contamination in the Dampier Archipelago, which could be used throughout the Indo-Pacific given its abundance in the region.