Imposex and tributyltin contamination as a consequence of the establishment of a marina, and increasing yachting activities at Phuket Island, Thailand

The objective was to investigate how tributyltin (TBT) contamination resulting from the establishment of a new marina and increasing yachting activities at Phuket Island, Thailand affects the area. A minimum of 30 specimen of the mangrove dwelling muricid Chicoreus capucinus were collected at five stations 2 months before and 5, 9, 18 and 33 months after Yacht Haven Marina was constructed. Imposex measured as RPLI and VDSI were significantly correlated to duration of time after construction and distance from the marina. All females developed imposex close to the marina after 18 months, whereas no significant increase occurred at stations more than 2 km from the marina. To evaluate the range of effects of the increasing yachting activities the muricid Thais distinguenda was used as an indicator of imposex at two popular mooring sites for yachts at Phi Phi and Raja Island in Phangna Bay, Thailand. The incidence of imposex was 100% at stations close to the mooring sites. A significant correlation existed between the distance from the sites and the incidence of imposex. The increasing incidence of imposex suggest that TBT contamination is worsening, against global trends, because regulations prohibiting the use of TBT-based paints, do not exist in Thailand.     

Ecological risk assessments of endocrine disrupting organotin compounds using marine neogastropods in Hong Kong

As active ingredients of anti-fouling paints that are widely used on ship hulls, organotin compounds, in particular tributyltin (TBT), are well-known endocrine disruptors causing sex changes in marine organisms and widespread in coastal waters and sediments worldwide. In this study, a comprehensive ecological risk assessment (ERA) of organotins was conducted in Hong Kong waters through determining the imposex status, sex ratio and tissue burdens of these compounds in the neogastropods, Thais clavigera and Thais luteostoma collected from 29 coastal sites. We also investigated the historical trend of organotin effects on these gastropods, and performed a probabilistic ERA based on tissue burden of TBT in the animals. Our results demonstrated that imposex indices were positively correlated with the body burden of organotins in the gastropods. Across all sites, the sex ratio (female:male) decreased significantly with increasing imposex levels or tissue burden of organotins, implying that such pollutants can result in a male-biased population, potentially leading to local extinction in extreme cases. Based on the ERA, 5.4% of all populations of T. clavigera are at risk due to exposure to TBT; the risks include growth inhibition, impairment of immune functions and reduced fitness. Seriously impacted areas included Aberdeen, Repulse Bay, Butterfly Beach, Mui Wo and Ha Mei Wan. A comparison with historical data revealed that there had been some improvement in the areas with low marine traffic, and distant from the major harbour/port. This could partly be due to the restriction on the use of TBT on small vessels (<25m in length) since 1992. Nevertheless, the organotin contamination still remains severe in areas with high marine traffic or adjacent to large harbours/ports. In particular, the situation in the northeastern waters of Hong Kong has been getting worst since 1996 that is probably associated with the rapid development of the cargo container port at Yantian in China.     

Quantification of organotin compounds and determination of imposex in populations of dogwhelks (Nucella lapillus) from Norway

Unintentional effects of the antifouling agent tributyltin (TBT) have been discovered in marine gastropods as the induction of male sex characters in females, known as imposex. The occurrence of imposex in dogwhelks (Nucella lapillus) was investigated in 41 populations sampled in 1993-1995 along the Norwegian coast. A method for quantification of organotin species using gas chromatography and atomic emission detection (GC-AED) was developed. Some degree of imposex occurred in almost all populations of dogwhelks studied, except in four from Northern Norway. The concentration of organotin compounds in the gastropods from the unaffected populations was below the detection limit (7 ng Sn/g d.w.). The concentration of TBT in dogwhelks from affected populations was in the range 48-1096 ng Sn/g d.w. A positive relation between the concentration of TBT in dogwhelks and the degree of imposex was found.     

Kinetics of the development of imposex in transplanted adult dogwhelks, Nucella lapillus

The degree of imposex in dogwhelks, Nucella lapillus (L.), is normally described in terms of a series of vas deferens development stages. The progression of adult female dogwhelks through vas deferens stages 0-5 in response to exposure to tributyltin at 2-128 ng/l has been modelled on the basis of first-order kinetics. Modelling indicated that the half-lives of stages 0, 1 and 2 were from 1-8 weeks and that the half-life of stage 3 was approximately twice as long. The half-life of stage 4 was 90-120 weeks. Half-lives decreased with increasing exposure concentration. The implications of these data for field surveys and transplantation experiments are discussed. Transplanted dogwhelks will not rapidly attain equilibrium with environmental conditions. Experiments will require careful control of exposure period, and preferable supporting chemical data.     

Organotin concentrations in three intertidal neogastropods from the coastal waters of Taiwan

This study was aimed to determine organotin concentrations in the muricids Thais clavigera, Thais tuberosa and Morula granulata on an island wide scale and to find an adequate bioindicator species for long-term monitoring purpose. Samples were collected from the coast of Taiwan and vicinity islands with organotin analysis by graphite furnace atomic absorption spectrophotometry. Because triphenyltin (TPT) could not be discriminated from tributyltin (TBT) by the extraction method, organotins were measured as TBT + TPT and total hexane extractable organotins. Severity of imposex in the muricid population samples expressed as relative penis size (RPS) indices were calculated and compared to determine a suitable bioindicator species. In T. clavigera, T. tuberosa and M. granulata, TBT + TPT were 17-157, 1-44 and 117-1343 ng Sn g(-1) dry wt, respectively. And total organotins varied in the range of 181 to 1125, 23 to 44 and 229 to 1402 ng Sn g(-1) dry wt, respectively. A positive correlation was found between TBT + TPT and total organotins in M. granulata (TBT + TPT = Total organotins x 1.01-110.79; R2=0.97; p<0.001). At the site of Dapaisha, total organotins in M. granulata was 10 times higher than in T. tuberosa (i.e. 242 versus 23 ng Sn g(-1) dry wt). No signs of imposex were found in T. tuberosa (a single site in two subsequent years) and in M. granulata (seven collection sites). The degree of imposex (RPS) of T. clavigera varied from 0.2 to 38.1%. The RPS indices from fishing harbors were not higher than from rocky shores and oyster culture sites. No correlation was found between RPS indices and concentrations of TBT + TPT or total organotins. M. granulata seemed to be insensitive to organotin pollution although organotins were also detected. Our results indicate that organotin pollution is wide spread in coastal waters of Taiwan and pollutants at least include butyltins and phenyltins. As a bioindicator species, T. clavigera is more appropriate than T. tuberosa and M. granulata to monitor organotin contamination.     

The recovery of populations of dogwhelks suffering from imposex in the Firth of Forth 1987-1997/98

The impact of tributyltin (TBT) contamination on dogwhelk (Nucella lapillus) populations was assessed at 33 sites in the Firth of Forth, UK, during spring 1997 and summer 1998. The sex ratio of the animals; the ratio of juveniles to adults; the degree of imposex, as determined by the relative penis size index (RPSI); and the total tin concentration in a sample of dogwhelk tissue were used as measurements of TBT impact on dogwhelk populations. These data were compared with data from a similar survey carried out at the same sites in the Firth of Forth in 1987 (Bailey, S.K., Davies, I.M., 1988. Tributyltin contamination in the Firth of Forth (1975-1987). Science of the Total Environment 76, 185-192.) before restrictions were introduced on the use of TBT. The results demonstrate a general recovery in dogwhelk populations from the impact of TBT at the majority of the sites studied, although at six of the sites, the RPSI value remains high enough to suggest breeding problems in the population. It is suggested that large vessels are responsible for localised cases of imposex within the Firth of Forth.     

Imposex occurrence in marine whelks at a military facility in the high Arctic

Imposex was found in the Arctic whelk Buccinum finmarkianum at several marine stations off Thule Air Base, an US military facility in Northwest Greenland. This indicates a widespread contamination with the antifouling agents, tributyltin (TBT) or triphenyltin (TPhT) in the area, but such contamination was not supported by the organotin analyses in sediments, whelks and clams, which in general was below the analytical detection limit. Organotin concentrations above the detection limit were found only at one station close to a quay, where the highest frequency of imposex also occurred. This suggests that imposex in B. finmarkianum is a biomarker of TBT more sensitive than the detection limits, which the analytical chemistry could achieve in this study.     

Tributyltin accumulation and effects in marine molluscs from West Greenland

The levels of the antifouling agent tributyltin (TBT) and its breakdown products in bivalves were investigated in 1999-2000 at six areas along the west coast of Greenland with focus on locations inside and outside harbours. In addition female gastropods were examined for the development of TBT-induced masculine characteristics in form of imposex or intersex. The highest TBT concentration, 254 ng x g(-1) ww, was found in the bivalve Mytilus edulis sampled inside Nuuk harbour, but significant TBT concentrations were also present in bivalves from the other harbour areas. Only low levels of TBT were detected in bivalves sampled outside the harbours and in several of the samples the TBT level was below the detection limit. The examination of neogastropods like Buccinum revealed that imposex development occurred in all the harbours. In contrast, imposex was not found in any neogastropods sampled outside the harbour areas. However, the value of marine neogastropods as indicators of TBT contamination in West Greenland seems limited, because of large species diversity and the difficulties in sampling enough specimens at least with the current sampling strategy. No effects, which could be related to TBT contamination, were found in the most abundant tidal gastropod in West Greenland, Littorina saxatilis.     

Assessment of imposex and butyltin concentrations in Gemophos viverratus (Kiener, 1834), from São Vicente,Republic of Cabo Verde (Africa)

This work constitutes the first assessment of tributyltin (TBT) pollution levels in the Republic of Cabo Verde (Africa) and proposes the marine gastropod Gemophos viverratus (Kiener, 1834) as a new bioindicator of TBT pollution in the Macaronesia and west coast of Africa. Specimens were collected between August and October 2012 along a gradient of naval traffic in São Vicente Island. The results clearly indicate an increase of imposex levels (percentage of females affected with imposex, 0–100 %; vas deferens sequence index, 0–4.1; relative penis length index, 0–54.6 %) and female TBT contamination (from 5 to 37 ngSn g^?1 dry weight (dw)) from outside to inside the harbour of Porto Grande Bay and identify this area as the focus of TBT pollution in the island. The butyltin degradation index for G. viverratus tissues ranged between 1.3 and 2.2, which being above 1 suggests that a considerable part of TBT inputs to the bay may not be very recent. Sterile females were found inside the harbour with an incidence up to 21.4 %. Considering the existence of a planktonic veliger stage in the life cycle of G. viverratus, it is expected that recruitment of newborn individuals can be supplied from unaffected breeding females inside and outside the Porto Grande Bay, resulting in a reduced impact of TBT pollution on population abundance. G. viverratus is very promising to be used as a simple, inexpensive and efficient novel tool for TBT pollution biomonitoring in the Macaronesia and west coast of Africa, a region for which there is an astonishing lack of information concerning levels and ecological impacts of TBT pollution.     

Organotins and imposex in the rock shell, Thais clavigera, from oyster mariculture areas in Taiwan

The amounts of organotin compounds such as butyltins [sigma BTs: tributyl- (TBT), dibutyl- (DBT), monobutyl- (MBT)] and phenyltins [sigma PhTs: triphenyl- (TPhT), diphenyl- (DPhT) and monophenyl- (MPhT)] in rock shells, Thais clavigera, from the northern (Shiangsan), central (Lukang) and southern (Chiku) Taiwan oyster mariculture areas varied with season and location. High values of sigma BTs (mainly TBT) and low values of sigma PhTs (mainly TPhT) were found during winter (January 1999) at Shiangsan, while high amounts of sigma PhTs (mainly TPhT) and low sigma BTs (mainly TBT) were found during summer (August 1998) at Lukang. The snails were also affected by imposex. Imposex at Shiangsan, Lukang and Chiku increased from 67.1, 59.3 and 36.7% in summer to 100, 100 and 80% in winter, respectively. Relative penis size indices similar trend at Chiku site. No male displayed rock shells were found at Shiangsan and Lukang in winter. Linear correlations between pseudopenis length versus TBT (r = 0.7655, P < 0.001), DBT (r = 0.4253, P < 0.05), MBT (r = 0.5865, P < 0.01) and TPhT (r = -0.6160, P < 0.01) were obtained significant. Among 200 samples, significant positive correlations between length (pseudopenis/shell) ratio of female versus TBT (r = 0.6944, P < 0.005) and sigma BTs (r = 0.6413, r < 0.01) were also observed. The weak correlations between length (pseudopenis/shell) ratio of female versus DBT (r = 0.3085) and MBT (r = 0.4240, P < 0.1) and TPhT (-0.4917, P < 0.05; negative correlation) could indicate that DBT, MBT and TPhT have little or no effect on the development of imposex in rock shells.     

Three decades of tributyltin in the coastal environment with emphasis on Arcachon Bay, France

This account briefly reviews the fate, bioconcentration and effects of tributyltin (TBT) three decades after its introduction into the coastal environment. Data on TBT-related events that occurred in Arcachon Bay (gastropod imposex, oyster recruitment failure, oyster shell malformations, concentrations in water and sediments) are revised and reinterpreted. Historically, TBT concentrations in the waters of the open bay seldom exceeded 1 ng Sn l(-1) in the 1960s, likely surpassed the range of 100 ng Sn l(-1) from 1977 to 1981 (years when no oyster spat were collected and the local shellfish industry collapsed), and decreased to about 1 ng Sn l(-1) in the late 1980s and early 1990s following legislation in January 1982 banning TBT use. TBT pollution is discussed in relation to the green tides that have developed in Arcachon Bay since 1982.     

Tributyltin contamination around an oil terminal in Sullom Voe (Shetland)

This report provides clear evidence of tributyltin (TBT) contamination of coastal waters close to an oil terminal handling large tankers. Dogwhelk populations from Sullom Voe, Shetland, have been examined for the development by females of certain male sexual characteristics (e.g. a penis and vas deferens). This condition, termed imposex, is a response to exposure to tributylin compounds, and can be quantified by use of the relative penis size index (RPSI). Dogwhelk populations throughout Sullom Voe were strongly affected by TBT (RPSI 34 to 81%), and up to 90% of the females in the oil terminal area were sterile. Queen scallops collected in the Voe contained enhanced concentrations of tin in adductor muscle and gonad tissue (0.02 to 0.23 mg kg(-1) wet weight), but it is considered unlikely that their growth and development is adversely affected. Tin was rarely detected in scallops, queens, or dogwhelks outside the Voe, although the development of a vas deferens in many of the female dogwhelks indicated a degree of contamination of the open waters of Yell Sound. TBT was detectable (2 ng litre(-1) as tin) in only two sea water samples taken from the terminal area.     

Imposex in the commercial snail Bolinus brandaris in the northwestern Mediterranean

A survey of the degree of imposex in populations of the commercial muricid Bolinus brandaris (L.) was carried out at six locations along the Catalan Coast (NW Mediterranean). Imposex was found to be a widespread phenomenon; the occurrence of a penis was observed in all females studied with the exception of those from St. Carles (South of Catalunya) where only 37% were affected. Other indices such as Relative Penis Length (RPL) and Vas Deferens Sequence (VDS) were applied; RPL varied from 4.5% in St. Carles--the least affected area--to 29.8% in the vicinity of Barcelona; similar results were obtained for VDS. Tissue organotin residues ranged from 6 to 140 ng g(-1) w.w. as Sn. Despite legislation restricting the use of tributyltin (TBT) in antifouling paints, this paper evidences the occurrence of organotin pollution far from the source, with levels of both TBT and triphenyltin (TPhT) high enough to cause environmental concern.     

Organotin contamination in the Atlantic Ocean off the Iberian Peninsula in relation to shipping

Imposex in female snails, a bioindicator of TBT contamination, and the presence of organotins in snails' tissue and sediments were studied at nine sites off the western Iberian Peninsula. The study was part of a European project (acronym HIC-TBT) co-financed by the EU-LIFE programme, intending to investigate and communicate the impact of organotins from ships in marine ecosystems. Snails and sediments were sampled during two cruises in May/June 1999 and in January 2000 in areas of high, intermediate and low-shipping density. Imposex was found in female snails from several sampling sites, some of which had an imposex incidence of 100%. Differences in sensitivity were found between species; hence comparison of imposex levels between locations where different species were collected was not straightforward. Total organotin concentrations in sediments (sum of butyl and phenyltin compounds) ranged from 21 to 185 ng Sn g(-1) with higher values for most sites sampled in the vicinity of shipping lanes. Organotin concentration in snails' tissue ranged from <5 to 196 ng Sn g(-1), which are similar to those found in snails from other offshore areas contaminated by TBT. Overall, these results give further support to the recent ban on the use of organotin based antifouling paints to all ship size.     

Trace elements and butyltins in a Dall's porpoise (Phocoenoides dalli) from the Sanriku coast of Japan

Concentrations and body burdens of 14 trace elements (Hg, Cr, Mn, Co, Cu, Zn, Sr, Ag, Cd, V, Se, Pb, Mo, and Fe) and butyltins (BTs) (tributyltin TBT, dibutyltin DBT, and monobutyltin MBT) were determined in various tissues of a mature male Dall's porpoise (Phocoenoides dalli) collected off the Sanriku coast of Japan. Selective accumulation in this porpoise was observed for Hg, Mn, Cu, Ag, Mo, Fe, and total BTs (TBT, DBT, and MBT) in the liver, Cd in the kidney, Zn, Sr, V, Pb, and Co in the bone, and Se in the skin. In contrast, Cr concentrations in all tissues were similar. This distribution pattern in this mature porpoise was in general agreement with the accumulation characteristics of trace elements and butyltins reported for other marine mammals. The whole body of the porpoise contained approximately 62 g Fe, 8.8 g Zn, 4.0 g Sr, 0.6g Se, 0.41 g Cu, 0.19 g Hg, 0.17 g Cd, 0.16 g Mn, 0.05 g Cr, 0.009 g Ag, 0.008 g Mo, 0.005 g Pb, 0.004 g Co, and 0.7 mg of BTs (0.4 mg TBT, 0.2 mg DBT, and 0.1 mg MBT). Metabolism of TBT to its breakdown products of this porpoise seems to be limited, since TBT still accounted for about half of the total burden of BTs. As in the cases of Hg, Mn, Cu, Se, and Fe, the muscle was the most important reservoir (43%) for the whole body burden of total BTs, 80% of which was TBT, and thus muscle played a crucial role in the higher body composition of TBT in this Dall's porpoise.     

Microbial degradation of bis(tributyltin) oxide

The degradation of the biocide bis (tributyltin) oxide by pur cultures of microorganisms has been studied. Attempts to isolate microorganisms able to utilize bis (tributyltin) oxide as sole carbon source, were unsuccessful. Of many bacterial species investigated $\text{Pseudomonas aeruginosa}$ and $\text{Alcaligenes faecalis}$ degraded sublethal amounts of bis (tributyltin) oxide during aerobic growth in the presence of suitable carbon sources. $\text{P. aeruginosa}$ converted bis (tributyltin) oxide into monobutyltin and small amounts of dibutyltin derivatives. Similar results were obtained with growing mycelium of $\text{Coniophora puteana}$, $\text{Trametes versicolor}$ and $\text{Chaetomium globosum}$. Microbial conversion of monobutyltin trichloride was not observed. Dibutyltin dichloride was converted into monobutyltin derivatives under certain sterile conditions.     

Butyltins concentration levels and imposex occurrence in snails from the Sicilian coasts (Italy)

Two sampling campaigns have been carried out along the Sicilian coasts within the frame of a European project (Hic-TBT) financed by the LIFE programme. Snails and sediments have been sampled at 5 stations, characterised by very low (Ustica natural sea reserve), low (Ustica harbour), intermediate (Castellamare and Termini Imerese) and high shipping densities (Palermo). Biological and chemical analyses were carried out in order to evaluate the incidence of imposex occurring in snails, and the concentration levels of organotins in sediment and snail samples. Despite the low organotin concentration levels in sediment and snail samples, the results show that imposex is widespread around Sicily. In four locations almost all female snails were affected by imposex at advanced development stages, even in areas with a fairly low shipping density. Only in the natural sea reserve of Ustica Island imposex was relatively less severe. Finally a logarithmic correlation between TBT and imposex stages was observed.     

Histopathological effects of chronic aqueous exposure to bis(tri-n-butyltin)oxide (TBTO) to environmentally relevant concentrations reveal thymus atrophy in European flounder (Platichthys flesus)

Although the use of tributyltin in antifouling paints has been banned, this compound is still a serious pollutant of the marine environment. This paper describes a unique study in which European flounder (Platichthys flesus) were chronically (8 months) exposed to bis(tri-n-butyltin)oxide (TBTO) in the water under controlled laboratory conditions. Residue levels in selected tissues (liver, muscle) and general health status indices were measured and the effects on several organs (gills, liver, mesonephros, ovary/testis, spleen, and gastrointestinal tract) were examined histopathologically. Additionally, morphometric analysis of the thymus was performed. The major finding is that exposure of flounder to 5 μg TBTO/l over a period of 8 months, resulting in body burdens comparable to high field levels, induced significant reduction of thymus volume, possibly affecting immunocompetence of the animals. Chronic exposure of European flounder to tributyltin is therefore likely to affect the general health status of this species in heavily polluted aquatic environments.     

Occurrence of organotin compounds in house dust in Berlin (Germany)

In a study in the year 2000 on the occurrence of hazardous environmental contaminants house dust samples from 28 Berlin apartments were measured for the presence and concentrations of six organotin compounds, monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), monooctyltin (MOT), dioctyltin (DOT) and triphenyltin (TPT). The concentrations of MBT and DBT determined ranged considerably from 0.01 mg kg-1 to 1.5 mg kg-1 (median: 0.05 mg kg-1) and 0.01 to 5.6 mg kg-1 (median: 0.03 mg kg-1), respectively. Maximum levels of TBT and MOT were only 0.08 mg kg-1 and 0.04 mg kg-1. The maximum total value of the organotins was 7.2 mg kg-1 (median: 0.11 mg kg-1). MBT was found in 86% and DBT in 82% of the samples above the limit of quantification, TBT and MOT only in 50% and DOT in 43%. The focus of ecotoxicology is on the risks arising from organotin compounds (especially butyltins) when used as biocides in antifouling paints. TBT acts as an endocrine disrupter in animals, inducing masculinization (imposex) in female gastropods of different species by increasing testosterone levels. The most critical organ site in experimental animals is the cellular immune system, where lymphocyte depletion in the thymus and peripheral lymphoid tissues takes place. Our study does not provide data on the basis of which population exposure could be estimated; house dust containing harmful organotins could, however, under some conditions, become a relevant intake possibility for young children.