Imposex in marine gastropods may be caused by binding of organotins to retinoid X receptor

Organotin compounds have been widely used as antifouling paints for ships and fishing nets since the 1960s and have thus been released into marine environments. Aquatic invertebrates, particularly marine gastropods, are extremely sensitive to organotin compounds such as tributyltin (TBT) and triphenyltin (TPT) and undergo changes in sexual identity in response to exposure. This worldwide phenomenon is one of the worst consequences of pollution by man-made chemicals and has led to the ban of such compounds in antifouling paints in a number of countries, although organotin compounds still exist in the environment. So far, very low-concentrations of TBT or TPT have been shown to induce imposex (superimposition of male genitalia on female) in marine gastropods. Although the imposex induction mechanism has been controversial for many years, it was recently reported that TBT and TPT are potent and efficacious activators of retinoid X receptor (RXR), a member of the nuclear receptor superfamily. In this review, I discuss the involvement of RXR in the development of gastropod imposex. Physical and Chemical Impacts on Marine Organisms, a Bilateral Seminar Italy–Japan held in November 2004     

有机锡致海洋腹足类性畸变分子机制的研究进展

有机锡作为防污损生物附着添加剂曾被大量使用,在极低浓度下就能诱发腹足类发生性畸变现象,严重危害了海洋生态系统的平衡。虽然三丁基锡已被国际海事组织全球禁用,但目前报道的环境有机锡污染仍然非常严重,而且其致毒机制一直众说纷纭。首先,简述了腹足类性畸变及其在世界各个海域有机锡污染监测中的应用,重点综述了有机锡致海洋腹足类性畸变分子机制的3种假说:脊椎动物类型的类固醇激素假说、神经肽假说以及视黄酸X受体(retinoid X receptor,RXR)假说,另外,结合快速发展的分子生物学技术,探讨了研究腹足类性畸变分子机制的新思路和新方法。     

Speciation of organotin compounds in sediments of semi-closed areas along the Mediterranean coast of Alexandria

Organotin compounds were measured in sediments of four different semi-closed areas of the Mediterranean coast of Alexandria: the Eastern Harbour, Western Harbour, El-Max Bay, and Abu-Qir Bay. Due to the commercial trade activity inside the Western Harbour, in addition to the effect of wastes discharged from El Noubaria canal, it shows the highest concentrations of total tin (6.34 μg g^-1 dry weight), dibutyl tin (1.63 μg g^-1 wet weight), tributyl tin (0.33 μg g^-1 wet weight) and diphenyl tin (1.06 μg g^-1 wet weight) compared with other locations. The concentrations of TBT species showed the highest contents compared with DBT and DPhT compounds in all sampling areas. This trend might be due to the worldwide use of TBT not only as biocides in antifouling paints but also as preserving agents for wood, fungicides in agricultural activities, and heat and UV stabilizers of PVC, which results in a direct release of TBT into the water body, accumulation in aquatic fauna. There is also precipitation into sediments and a decrease in degradation rate into its derivatives. Variations, types, concentrations, and distribution of different organotin compounds are discussed in the areas under investigation. The study reveals a new record of organotin compounds along the Alexandria coast and makes comparisons with other surrounding areas of interest.     

Retrospektives Monitoring von Organozinnverbindungen in biologischen Proben aus Nord- und Ostsee – sind die Anwendungsbeschränkungen erfolgreich?

Background, aim, and scope Organotin compounds are used as biocides, plastic additives and catalysts. With respect to environmental effects, tributyltin (TBT) and triphenyltin (TPT) compounds are the most relevant, because of their high aquatic toxicity and endocrine effects on mussels and snails. TBT was mainly used as antifouling agents in coatings of ships and boats. In 1989, Germany banned the application to ships <?25?m length. Finally, in 2003, the use of organotin-based antifoulants within the European Union was completely banned. To verify the effectiveness of the restrictions a retrospective monitoring study was initiated. Material and methods A set of appropriate archived samples was retrieved from the German environmental specimen bank (ESB) comprising standardized pooled samples of eelpout (Zoarces viviparus) muscle tissue and of soft bodies of common mussels (Mytilus edulis) sampled at two locations in the North Sea and one in the Baltic Sea. Analysis of organotin compounds included n-hexane extraction, derivatisation with sodium tetraethyl borate, capillary gas-chromatographic separation and atomic emission detection. Results and Discussion Altogether, time series cover the period 1985 to 2006, including data of an earlier study (Rüdel et al. 2003). Until the late 1990s, TBT remained more or less constant in all samples (e.?g. 17?±?3?ng/g wet weight, ww, in mussels from Jadebay/North Sea). The German ban of TBT-based antifoulings for small ships had no effects on environmental concentrations because large ships dominate in the investigated North Sea regions. After the EU-wide ban of TBT in 2003, however, significant decreases in mussel and fish contamination could be observed. In mussels from Jadebay, TBT concentrations were 14 and 6?ng/g ww in 2004 and 2005, respectively. TBT contamination in eelpout of the same region decreased to about 30 percent of the initial concentrations in 2006. Corresponding decreases were detected for TPT. Declining trends were also found in fish and mussels sampled from a Baltic Sea offshore site. Conclusions The results demonstrate the effectiveness of the legal measures undertaken to control organotin inputs into the aquatic environment. Nevertheless, organotin compounds are still relevant pollutants. Water concentrations calculated from the measured tissue concentrations by using the respective bioconcentration factors are still above the Environmental Quality Standards derived in the context of the Water Framework Directive (0.2?ng/l) and the OSPAR mussel EAC (Environmental Assessment Criteria; 2.4?ng/g ww). Thus adverse effects to marine organisms cannot be excluded. Recommendations and perspectives Further studies should be performed to verify the declining trends. More sensitive analytical methods, e.?g. species-specific isotope dilution analysis, are recommended in order to detect lower environmental concentrations.     

Toxicological effects of mouse diet contaminated with tributyltin on the immune and neurological systems of C57BL/6 mice

Organotin compounds, widely used as antifouling agents, are known to bioaccumulate in the food chain. Among organotin compounds, tributyltin (TBT), a toxic and widespread contaminant, has become a serious factor in environmental pollution and is suspected of being immunotoxic in animals. The purpose of this study was to assess the effects of 80 µg of TBT per kilogram of body weight (kg bw), provided in food, on the immune and neurological systems of C57Bl/6 mice. Data showed that TBT increased the proliferation of T and B lymphocytes in both adult and juvenile female and juvenile male mice, but that it decreased the proliferation of both T and B lymphocytes in adult male mice. The macrophages activity in female and male juvenile mice was higher than in adults of both sexes. The natural killer cytotoxic activity was also increased in juvenile and adult males and females compared to the control groups. In the brain, we observed the presence of TBT in the hippocampus, the striatum, the cortex, and the cerebellum in both the male and female groups. The highest levels were observed in the cortex of females and males, while the lowest levels were found in the cerebellum. TBT also induced an increase in the levels of the antioxidant glutathione (GSH) in the striatum, the hippocampus and in cortical structures but not in the cerebellum where the levels of TBT are lower. Our findings indicate that TBT modulated the immune and nervous systems causing endocrine and nervous perturbations.     

Comparative effects of two antifouling paints on the oyster Crassostrea gigas

The effects of two antifouling paints, one containing organotin compounds, the other copper oxide, were studied from September 1983 to October 1984 on Crassostrea gigas (Thunberg) grown under natural field conditions in the Bay of Arcachon, France. The organotin paint reduced growth rate expressed as weight, length and width, but did not affect shell height; it drastically decreased the dry condition factor and shell density, but did not affect the viability of embryos and larvae from exposed oysters. However, some decrease in larval growth rate was observed. The copper paint had no effect on oyster growth, but lowered the condition factor compared to controls. Neither viability of embryos or larvae nor larval growth were affected by this paint.     

Ecotoxicity studies of some organotin compounds

The ecotoxicities of three organic compounds containing spacer groups, an intermediate organotin premonomer, four organotin monomers, their homopolymers and six copolymers of tributylin α‐chloroacrylate (TCA) are reported against two microorganisms viz., Pseudomonas aeruginosa and Sarcina lutea and a larval species Anemia salina (Nauplii II). The organic compounds containing spacer group were less toxic whereas the monomers were found to be highly toxic. The homopolymers were found to be more toxic than that of tributyltin acrylate (TBTAA), tributyltin methacrylate (TBTMA) and tributyltin α‐Chloroacrylate (TCA). The toxicity of TCA copolymers increased with the increase in tributyltin (TBT) content in the polymer. From this study it was not possible to establish any relationship between the chemical nature of the comonomer with its toxicity.     

Bioconcentration of butyltin compounds by round crucian carp

Bioconcentration factors (BCF) by round crucian carp and partition coefficients between n‐octanol and water (Pow) were measured for dibutyltin, tributyltin and triphenyltin compounds. Pow was 0.97–3.66 for 7 organotin compounds. BCF of tributyltin and triphenyltin compounds was 1.70–2.77 for muscle, 1.70–2.66 for vertebra, 2.05–3.70 for liver and 1.49–3.50 for kidney.     

Application of rotating cylinder method for ecotoxicological evaluation of antifouling paints

The environmental impact of two biocide-free antifouling paints, fluoropolymer and silicone types, painted on a test cylinder was assessed using a battery of ecotoxicity test and chemical analyses for organic micro-pollutants such as perfluoroalkyl substances (PFAS). A biocide paint containing zinc pyrithione (ZnPT₂) and cuprous oxide was assessed as a positive control. A standardized laboratory rotating-cylinder method using each test cylinder with artificial seawater was performed for 45 days. After 1?h rotation, the leaked seawater was subjected for bioassay and chemical analyses twice weekly. The seawater extracts from the biocide paint showed adverse effects on bacteria, algae, and crustaceans, but those from the biocide-free paints did not. The leakage seawater from biocide-free paints, after 7-day continuous rotation, contained the same concentration levels of PFAS as blank seawater used to conduct the tests. Thus, no significant toxicities of the biocide-free paints were found under the conditions of this study. Simultaneous analysis of ZnPT₂ and copper pyrithione (CuPT₂) was developed using an HPLC with a polymeric resin column, showing that ZnPT₂ was converted to CuPT₂ by trans-chelation in the leakage seawater from the positive control paint. The experimental results using a laboratory rotating-cylinder method demonstrated that biocide-free paints did not markedly affect three species tested and no PFAS was detected. In contrast, biocide paint was significantly toxic to test species and toxicity of the extract on bacteria was partly responsible for CuPT₂ produced in leakage seawater. Thus, a laboratory rotating-cylinder method may be applied for ecotoxicological assessment of antifouling paints.     

使用底栖和浮游桡足类生物测试评估沉积物修复方案的效果

三丁基锡（TBT）是一种常见于船舶防污涂料中的有机锡化合物。尽管该化学物已于2008年被禁用,但在ports和港口等海上交通区域仍可检出致毒水平（浓度）的TBT。一项补救方案在新西兰的一个港口实施,以将TBT和铜的浓度降低到可接受的（浓度）值。这项研究使用分析化学和桡足类生物测试相结合的方法评估了该方案的有效性。在港口内的三个空间梯度上采集沉积物和水样,并在修复前后测定了有机锡和金属含量。利用底栖和浮游桡足类生物测试估算了沉积物和淘析样品的毒性。虽然底泥样品的急性毒性在修复后减少,但对底栖桡足类的繁殖率仍有影响。本研究所采用的方法有望评估沿海地区修复过程的效果。     

Monitoring the effectiveness of the European tributyltin regulation on the Basque coast (northern Spain) by assessing imposex in two gastropod species (Nassarius reticulatus and Nassarius nitidus)

Imposex and butyltin body burden were assessed in 2011 along the Basque coast (northern Spain) in two gastropod species (Nassarius reticulatus and Nassarius nitidus) four years after an initial survey in 2007. The aim of this re-survey was to monitor the effectiveness of the European ban on the use of tributyltin (TBT) based antifouling paints on ships’ hulls (EC 782/2003). Imposex levels in 2011 were lower than those determined in 2007 at most of the sampling sites. Accordingly, TBT concentrations in the female body burden of Nassarius reticulatus varied from 43 to 250 ng Sn/g in dry weight in 2011, which was a lower maximum than in 2007. Nevertheless, the results for the butyltin degradation index suggest that there have been recent inputs of TBT within the two main Basque harbours. Overall, the legislative measure is contributing to the reduction of TBT effects on the Basque coast although its presence is still evident.     

Marine succession of fouling organisms in Hong Kong,with a comparison of woody substrates and common,locally-available,antifouling paints

Successional stages of macrofaunal fouling organisms of test blocks were exposed in the subtropical waters of Hong Kong. A comparison of 5 types of wood commonly used to build boats, and 5 types of locally available antifouling paints is made. Rafts holding the test blocks were exposed at 1 m depth from 24 March, 1970, to 23 October, 1970. The initial macrofoulers on the control (chinese fir) were Balanus and Bugula, and the final dominance was a Styela-Mytilus community. On painted surfaces, the Balanus-Elasmopus community was dominant. Temperature, salinity, dissolved oxygen, and turbidity were recorded weekly during the test. A preliminary chemical analysis of percentage pigment and cuprous oxide content in the antifouling paints is presented.     

Comparison of heavy metal concentrations in fish samples from three fish farms (Eastern Mediterranean) utilizing antifouling paints

Although aquaculture is considered the fastest growing food production industry, nevertheless there is little information regarding pollutant levels in cultured fish. Samples of cultured sea bass – Dicentrarchus labrax (Linnaeus, 1758), sea bream – Sparus aurata (Linnaeus, 1758), and sharpsnout sea bream – Diplodus puntazzo (Walbaum, 1792) from three fish farms located in the Eastern Mediterranean which utilized antifouling paints on the nets were analyzed for quantitative determination of zinc (Zn), chrome (Cr), and copper (Cu) in the gills, the liver, and the muscle separately for each tissue and fish. The results show that the highest levels of zinc (up to 29.6 mg kg^?1 dry wt) were found in the gills of all samples of sea bream and sharpsnout sea bream while in the samples of sea bass, the highest levels of zinc and chrome were found in the liver (up to 39.3 and 13.8 mg kg^?1 dry wt, respectively). Insofar, as copper is concerned the highest exposure was found in the liver of sharpsnout sea bream (up to 5.49 mg kg^?1 dry wt). The results indicate that the use of antifouling paints in aquaculture practices are a significant source of chemical pollution in cultured fish and poses a risk to the exposed organisms.     

Untersuchungen zur Nachhaltigkeit der Sanierung von Tributylzinnkontaminiertem Hafensediment durch Landablagerung

To allow ships to pass, most harbours have to be liberated from sedimented material. These sediments often contain high concentrations of tributyltin (TBT), which is used as a biocide in antifouling paints. Thus, the problem of a disposal which is acceptable with respect to a minimum risk for humans and the environment arises. The aim of our project was to assess land deposition as an alternative in dealing with TBT-contaminated harbour sediments. Therefore, we followed the biological degradation of TBT under aerobic and anaerobic conditions at different temperatures using defined laboratory experiments, and got important information about the land deposition, performing field measurements including biomonitoring. The biological degradation of TBT is faster under aerobic conditions and with increasing temperature. The half-lives found show the dependency of the degradation rates on the temperature between 5–55°C. The degradation rate determined for water under the same conditions (just biological degradation, without photolysis) is almost 4 times higher than in sediment. The field studies showed degradation rates of a maximum of 10–15% per year in untreated sediment and 30% per year in restacked sediments. We did not observe any interference of released TBT with ground water or surrounding areas. The TBT uptake in plants was low. Based on our results a multidimensional risk evaluation was performed, concerning the TBT and its degradation products dibutyltin (DBT) and monobutylin (MBT) released from the deposition areas. The land deposition appears to be a sustainable solution for dealing with TBT-contaminated harbour sediments.     

Tributylzinn auf dem Weg zu den ‘Top of the POPs’?

Two groups of pollutants established during the past years, the impact of which on the eco-system gave cause for concern and legislative reaction: Persistent Organic Pollutants (POPs) and Endocrine Disrupters. At present, only the 12 POPs (‘Dirty Dozen’) are subject to serious international bans as long as they do not belong as well to potential endocrine disrupters, such as the PCDDs/PCDFs (Polychlorinated Dibenzo(p)dioxins and furans). But there are endocrine disrupters which fulfil, the POP-criteria stated in the Stockholm Convention. The organotin compounds are part of them, especially the tributyltin (TBT), since the toxicity increases with increasing alkylation. The present investigations may contribute to the discussion on the POP-potential of TBT on the basis of the results from GfA-internal and literature findings.     

Some aspects of heavy metals contamination remediation and role of biosurfactants

Contamination by heavy metals is the result of different industrial activities. The presence of heavy metals in soil and water causes serious problems, as these materials are not biodegradable and do contaminate both biological systems and the subsoil. Biological surface-active compounds otherwise known as biosurfactants in general and rhamnolipids biosurfactants in particular have been successfully employed in the remediation of environments contaminated with heavy metal ions. The aim of the present review is to highlight potential applications of these tensioactive compounds for use in environmental heavy metals removal and bioremediation and processes involved.     

Hinia reticulata and Nucella lapillus. Comparison of two gastropod tributyltin bioindicators

Hinia reticulata and Nucella lapillus (collected from 1989 to 1991 at Pointe de Pléneuf and Méan Mélen, France, respectively) exhibit imposex in response to tributyltin (TBT) pollution leached from antifouling paints and proved to be good TBT bioindicators. H. reticulata was kept for 18 mo under TBT-free conditions in the laboratory, but no evidence for imposex remission was found. A second series of experiments showed that food chain uptake of TBT from a contaminated diet is an important mode of TBT exposure in H. reticulata. Comparative tank experiments with H. reticulata and N. lapillus demonstrated that the same type of TBT exposure resulted in comparable TBT body burdens, biological concentration factors, and imposex development in both species [measured as increase of VDS (vas deferens sequence index), uncubed RPS (relative penis size index) and average female penis length]. Differences in imposex development of natural and laboratory populations are discussed against the background of different types of TBT contamination of their food. A statistical study, based on an analysis of natural populations of both prosobranch species, makes a comparison of the specific TBT sensitivity of the two bioindicators possible. As a consequence it is proposed that TBT biomonitoring programs in Europe should use both prosobranchs as indicator species.     

Monitoring of heavy metals and butyltin compounds in coastal waters of Ligurian Sea: Results of a mussel watch project

From December 1994 up to October 1995 a “Mussel Watch Project”; was carried out to assess the quality of water of the Ligurian Sea and to identify the possible sources of heavy metal pollution. Several metals and organotin compounds were measured in mussels (Mytilus galloprovincialis, Lam) transplanted in different months from a marine farm to eight sites of the Ligurian coast.

Spatial differences and temporal trends in tissue concentrations of Zn, Cu, Ni, Cr, Fe, Al, V, Cd and Mn were evaluated. Concentrations resulted in the same range as those obtained in a previous Mussel Watch Project carried out in the same area. In general, areas of known pollution display high concentrations of these metals, but also some areas not greatly influenced by human activities showed high concentrations of some metals.

Data from this study show little evidence of any overall seasonal pattern except for Cu, Zn and V, concentrations of which were a function not only of sampling site, but also of reproductive cycle.

Organotin compounds show differences in terms of both concentration and speciation between the investigated sites.     

Marine bacterial fouling: A scanning electron microscope study

A range of substrates has been examined in order to determine the types of attachment mechanism employed by marine bacteria. Use of the scanning electron microscope (SEM) has also allowed an investigation of the initimate relationship between an antifouling paint matrix and its attached bacterial community. Plastic (Thermanox) and glass coverslips, together with Cu₂O-based and TPTF-based antifouling paints and their respective empty-matrix analogues have been used in this study. Observations over periods of up to 4 wk have shown that extensive bacterial communities can develop. A variety of bacteria have been found: cocci; various rods; stalked forms; and prosthecate types. These bacteria also exhibit a range of attachment mechanisms. Initial attachment is by simple stickiness of cell walls, flagella, pili (fimbriae) or stalks. This stickiness can be attributed to an actual adhesive, electrostatic forces, electrical double-layer phenomena or to London/van der Waals forces. Often, attachment is subsequently improved by the secretion of insoluble, high molecular weight, polysaccharidic material. This material is found in the form of strands, pads, sheets or capsules and acts to bridge the space between the cell wall and substrate or adjacent cells. Thus, stalked forms are found attached by basal pads of mucilage whilst cocci and rods are often found enmeshed in mucilage strands and sheets, but less often attached by pads and capsules. Initially, single bacterial cells attach and give rise to colonies by cell division. Continuing growth of adjacent colonies leads to the development of confluent films over the substrate surface. Further growth results in thickening until eventually the entire surface is coated with a dense community of mixed bacteria together with their adhesive mucilage. In the case of antifouling paints, the porous nature of the matrix allows invasion by bacteria and the possibility of pore blockage by adhesive polysaccharides. This problem is discussed with reference to the paint's loss of antifouling efficiency.