Accumulation of organotin in Littorina littorea and Mya arenaria from Danish coastal waters

Organotin concentrations were determined in seawater, sediment and selected molluscs collected from the vicinity of the island of Fyn, Denmark. The extent of organotin contamination varied with the proximity of sampling sites to marinas and commercial shipping activities. Seasonal reductions in environmental TBT concentrations coincided with removal of pleasure craft from marinas during the winter season. At marina sites, concentration factors for TBT were 150-1500 for sediment and 500-10,000 for Littorina littorea (compared with seawater concentrations). Away from marina sites, Mya arenaria concentrated TBT by a factor of 57,000 to 220,000. The concentrations of TBT in M. arenaria from relatively clean sites were more than 10 times higher than found in L. littorea from heavily polluted marinas. M. arenaria were absent from marina sites and their immediate surroundings. Of the total organotin found in M. arenaria throughout the year, 80-90% was in the form of TBT. In contrast, in L. littorea approximately 60-70% of the total organotin was present as TBT in spring, but this proportion fell to 20-40% during autumn and winter. M. arenaria shows potential as a bioindicator organism of TBT pollution as it appears to have a very limited ability to metabolise and eliminate TBT, yet can tolerate quite high levels of TBT exposure. L. littorea might be used as a bioindicator of short-term fluctuations in TBT concentrations at heavily polluted sites. It is concluded that continued presence of TBT in the Danish coastal ecosystem may still pose an ecotoxicological threat.     

Distribution of organotin compounds in tissues of mussels Mytilus edulis and clams Mya arenaria

Concentrations and composition of butyltin (BT) and phenyltin (PT) compounds were compared in tissues of mussels Mytilus edulis and clams Mya arenaria collected from the Mersey Estuary. Tributyltin (TBT) concentrations were consistently highest in digestive gland and low in muscular tissues such as siphons (Mya) and adductor muscle (Mytilus). Lowest concentrations were found in the periostracum surrounding the siphons of Mya. Ratios of monobutyltin (MBT), dibutyltin (DBT) and TBT to total BT content of Mytilus tissues were in the ranges 25-34%, 21-28% and 44-51%, respectively. Much higher proportions of TBT:total BT were characteristic of Mya (>80% in the digestive gland, remaining tissue and gonad) implying that slow degradation rates account for the relatively high levels of TBT in this species. TPT was the only phenyltin compound detected (albeit at relatively low levels), and, like TBT, was present in highest concentrations in the digestive gland of clams--suggesting an important dietary component in the accumulation of OTs in these bivalves.     

Biochemical responses in Salmo salar muscle following exposure to ethynylestradiol and tributyltin

This study investigated the effects of nominal concentrations of two endocrine-disrupting chemicals (EDCs) on fish physiology. The effects of the synthetic pharmaceutical estrogen ethynylestradiol (EE(2)) and the antifoulant tributyltin (TBT) were investigated in exposure studies with immature Atlantic salmon, Salmo salar. Fish were exposed for 7 days to waterborne EE(2), TBT, or a combination of both. The activities of lactate dehydrogenase (LDH), acetylcholinesterase (AchE) and glutathione S-transferase (GST), and lactate and glycogen content were determined in samples of fish muscle. Fish exposed to EE(2) for 3 days responded rapidly with increasing AchE and GST activities and increasing lactate content. These responses were limited to the first 3 days of exposure and had disappeared by day 7, indicating that the fish had adapted to EE(2) exposure. Compared to the controls, TBT increased AchE and LDH activity, inhibited GST activity and had no effect on lactate content. When mixed, the highest concentration of EE(2) increased the effect of TBT on lactate content. However, fish exposed to a lower concentration of EE(2) in combination with TBT had the lowest lactate content. Effects on AchE and LDH activities were smaller when TBT was combined with EE(2) compared to TBT alone. This suggests that TBT and EE(2) influence biochemical processes in fish muscle, acting on different organizational levels, by antagonistic and synergistic mechanisms.     

Biodegradation capacity of tributyltin by two Chlorella species

Two microalgal species, Chlorella vulgaris and Chlorella sp., which showed high tributyltin (TBT) tolerant ability were investigated for their capabilities in degrading TBT at sublethal concentration. The distribution of TBT and its degraded products dibutyltin (DBT) and monobutyltin (MBT) in the incubation medium, extracellular surface and intracellular fraction were monitored during an exposure period of 14 days. Results showed that biosorption of TBT by the algal cell wall was the major mechanism in reducing 40% of the initial TBT from the medium in the first 2 days. The half-life of TBT incubated with C. vulgaris was 60 h while that of Chlorella sp. was 80 h. The occurrence of DBT at Day 1 in the culture medium provided direct evidence to the biodegradation of TBT by both Chlorella species. At the end of the experimental period, 27 and 41% of the original TBT were recovered as DBT and MBT in cultures of C. vulgaris, respectively. In contrast, DBT appeared to be the only degradation product of Chlorella sp. and only 26% of the original TBT was transformed to DBT. Despite the same genus, TBT was debutylated to a greater extent to MBT by C. vulgaris, while DBT was the end degradation product by Chlorella sp. The capability of such debutylating process therefore accounted for the higher tolerant ability of C. vulgaris than Chlorella sp.     

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.     

Preliminary investigation of a sensitive biomarker of organotin pollution in Chinese coastal aquatic environment and marine organisms

In nine batches of sea bivalves collected from Chinese coastal cities during the year of 2000 to 2002, a special sample named Mya arenaria was found to have strong ability of butyltin accumulation compared with the other sampled bivalves in the corresponding batches. Tributyltin compound was the predominant pollutant with the detection rate high up to 100%. Special high levels of g Sn/g were detected in some Mya arenaria samples. The results obtained showed that Mya arenaria was potentially a biomarker to indicate organotin pollution in coastal aquatic environment.     

Effect of tributyltin on veliger larvae of the Manila clam, Ruditapes philippinarum

We investigated the effects of waterborne and maternal exposure to tributyltin (TBT) on veliger larvae of the Manila clam, Ruditapes philippinarum. In a waterborne exposure test, veliger larvae (D-larvae stage: 24h after fertilization) were exposed to TBT at measured concentrations of <0.01 (control), 0.055, 0.130, 0.340, and 0.600microg/l for 13d. The percentage of normal veliger larvae (the ratio of normal veliger larvae to all larvae) decreased significantly in all TBT treatment groups compared with that in the control group. In a maternal exposure test, 100 clams were exposed to TBT at measured concentrations of <0.01 (control), 0.061, and 0.310microg/l at 20-22 degrees C for 3 weeks, and the percentage of normal veliger larvae assessed for 13d. No maternal effects on veliger larvae from TBT were observed in TBT treatment groups as compared with the control group. These results demonstrate that waterborne TBT affects Manila clam veliger larvae, and indicates that TBT may have reduced Manila clam populations by preventing the development and survival of veliger larvae.     

Differences in Cd elimination from Mytilus californianus and Mytilus trossulus soft tissues

Field results have shown that Mytilus californianus is able to release its Cd concentrations significantly in just a few days. The existing paradigm states that Cd elimination from Mytilus soft tissues is a very slow process. This discrepancy was investigated in the laboratory, testing the effect of two Cd levels (10 and 1 microgram l-1) on its release from Mytilus trossulus and M. californianus soft tissues. After exposure to 10 micrograms l-1, both species showed a significant uptake with no elimination after several days of depuration. After exposure to 1 microgram l-1, the responses were different. No significant Cd uptake was seen in M. trossulus while in M. californianus uptake was significant but returned to the background level after just 1 day of depuration. This response of M. californianus is consistent with that reported from field studies. These results are important for environmental monitoring programs since M. californianus has been used as equivalent to other Mytilus species in the assessment of Cd pollution.     

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.     

Transplacental transfer of butyltins to fetus of Dall's porpoises (Phocoenoides dalli)

Here we provided evidence, for the first time, on the placental transfer of butyltin species in Dall's porpoise (Phocoenoides dalli). The maternal to fetal transfer of butyltins including monobutyltin (MBT), dibutyltin (DBT) and tributyltin (TBT) was investigated in a mother-fetus pair collected off the Sanriku coast of Japan. The maternal individual was found contaminated as its liver accumulated the highest concentrations of all MBT, DBT and TBT. This study revealed that the transfer rate was about 0.3% for total butyltins to a fetus of about 6 months old. Unlike in the mother, the fetal liver does not appear to be a preferential organ for accumulation of all the butyltins. In a approximately 100 kg-weight mother Dall's porpoise and its fetus of 2.21 kg-weight about 1,269 microg (TBT, 578 microg; DBT, 480 microg; MBT, 211 microg) and 4.2 microg of total butyltins (TBT, 1.1 microg; DBT, 2.1 microg; MBT, 0.96 microg) were found, respectively.     

Multi-tolerance to heavy metals in Plantago arenaria Waldst. & Kit.: adaptative versus constitutive characters

Tolerance to Cu, Cd, Ni and Zn was investigated in a population of the pioneer species Plantago arenaria growing in a metallurgical landfill. Tolerance levels were compared with those of two other pioneer species (Coniza sumatrensis and Verbascum densiflorum) growing in the same location, and with a control population taken from an uncontaminated site. Results showed that the metalliferous population of P. arenaria was more tolerant to metal toxicity than C. sumatrensis and V. densiflorum. Comparisons with literature data confirmed that the metalliferous population of P. arenaria was highly tolerant to Cu, moderately tolerant to Cd and Ni, but not particularly tolerant to Zn. The control population of P. arenaria responded the same as the metalliferous one excepted for Cu, for which it was much more sensitive. This suggested that multi-metal tolerances in the metalliferous population of P. arenaria resulted both from constitutive and adaptative traits, depending on the metal. To check whether P. arenaria was able to cope with high internal metal levels, accumulation patterns were evaluated in pot experiments. Results showed that metals accumulated in roots and leaves, at levels proportional to soil content. Metal content was much higher in roots than in leaves and the leaf:root concentration ratio was kept constant over a wide range of soil metal contents. This suggested that metal tolerance was related to the ability to retain metal ions in roots and to tightly control their translocation to leaves. Finally metal tolerance in P. arenaria is discussed in relation to its pioneer and xerophytic characteristics.     

Butyltin compounds in sediment and molluscs from the shipping strait between Denmark and Sweden

The distribution of tributyltin (TBT) contamination in the subtidal zone of the waters between Denmark and Sweden was investigated in relation to major international shipping lanes. Sediment and different benthic molluscs were sampled in transects along and perpendicularly to the shipping lanes in the Sound (?resund) and the Kattegat/Skagerrak region. The samples were analysed for TBT and its degradation products, dibutyltin (DBT) and monobutyltin (MBT), using GC-PFPD. In sediments, the TBT concentration ranged from <1 to 19 ng g(-1) dry weight (dw) with a strong correlation between the TBT concentration and the organic fraction in sediment (r(2)=0.90) in the samples collected in the Sound, where the highest concentrations were found. This relationship was not observed in the samples from the Kattegat because the TBT concentration in most sediment samples was below the limit of detection. In the molluscs, TBT and its degradation products were detected in all samples from the entire area with concentrations ranging from 8.1 ng g(-1) dw in Buccinum undatum to 1316 ng g(-1) dw in Nuculana pernula. The deposit-feeding bivalve N. pernula was found to have a particularly high accumulation potential for TBT. In addition, a strong correlation between TBT concentrations in sediment and N. pernula was found. Therefore this species seems to be an ideal organism for monitoring sediment contamination. The TBT concentration in N. pernula was found to decrease gradually along the shipping lanes from the Sound, through the Kattegat and into the Skagerrak.     

Factors influencing organotin distribution in different marine environmental compartments, and their potential health risk

Tributyltin (TBT) and triphenyltin (TPT) in different marine environmental compartments such as seawater, sediments, and inshore fishes were investigated in 21 Taiwanese harbors between 2001 and 2004 in order to determine the major factors influencing their distribution. The existence of major input sources and the limited water exchange rate inside the harbors were indicated by higher TBT concentrations in seawater from inner harbor than from outer harbor areas. The levels of TBT in sediments were found to be mainly affected by their geographic distribution, water exchange rates and shipping activity. No significant correlations in TBT concentrations between water, sediment and fish suggested TBT accumulation by fish might not result from water and sediment, but from their food. TPT were detected in most fish samples, but found in few sediment samples and none in seawater, indicating fish could be as a target element for monitoring contaminated levels of TPT in the aquatic environment. Mean concentrations of TBT in fish muscle higher than tolerable average residue levels (TARLs), and mean hazard indices of TBT and TPT higher than 1 suggested consumption of fishes from Taiwanese harbor areas might have potential high risk to human health.     

Effect of TBT on Ruditapes decussatus juveniles

The effects of sublethal concentrations of tributyltin (TBT) on growth of juvenile clams Ruditapes decussatus were determined during exposure to TBT concentrations of 50, 100 and 250 ng l(-1) (as Sn) for a period up to two years. Length and weight of clams increased continuously in all treatments throughout the experimental period, and, overall, rates were not significantly influenced by TBT exposure, although final length and weight were inversely related to increasing TBT concentration. Juvenile R. decussatus therefore appear to be less sensitive to TBT than larval stages. Some juveniles exposed to TBT developed abnormal shell growth, laterally, changing the typical flattened shape of clams into a more "rounded" form. This characteristic was more visible in the anterior margins of valves than posteriorly, and mainly observed in clams exposed to TBT at 50 ng l(-1) (as Sn).     

Impact of chronic cadmium exposure at environmental dose on escape behaviour in sea bass (Dicentrarchus labrax L.; Teleostei, Moronidae)

The effect of chronic exposure to a low concentration (0.5 microg l(-1)) of cadmium ions was investigated on escape behaviour of sea bass, Dicentrarchus labrax, using video analysis. Observations were also performed on the microanatomy of lateral system neuromasts. When fish were exposed for 4h per day over 8 days to the cadmium ions, most of both types of neuromasts observed remained intact. However, some of them presented damaged sensory maculae. Whereas before cadmium exposure, fish responded positively to nearly all the lateral system stimulations, after exposure they decreased by about 10% their positive responses to stimulations. From the 15th day after the beginning of cadmium exposure, neuromasts presented progressively less damage, cadmium accumulation in gills and scales decreased significantly and fish escape behaviour had recovered. This study presents a new concept in ecotoxicology: using behavioural change to reveal the effects of pollution levels, scarcely detectable by currently used techniques (physiological responses).     

The use of Littorina littorea for tributyltin (TBT) effect monitoring - Results from the German TBT survey 1994/1995 and laboratory experiments

Field investigations and laboratory experiments were performed to test the applicability of the intersex phenomenon in Littorina littorea for biological TBT-effect monitoring and to give further insights into toxicological aspects of intersex development. The intensity of the phenomenon is correlated with the degree of TBT contamination in the environment. Statistically highly significant correlations of biological indices and TBT concentrations in tissue and sediment exist. Intersex can be induced only in juvenile and sexually immature females and its intensity depends on the ontogenetic stage of development during TBT exposure. For the first time, malformations in male periwinkles closely related to the TBT contamination can be described; the reduction of male mamilliform penial glands shows highly significant correlations to TBT concentrations in tissues. The intersex phenomenon in Littorina littorea is proposed for a biological TBT-effect monitoring in those areas where the dogwhelk Nucella lapillus, as the more sensitive species in European surveys, is absent. The intersex index (ISI) as the mean value of the intersex stages in a sample should be used as the most sensitive biological parameter for the assessment of the TBT contamination in Littorina littorea.     

Changes in cell-type composition in digestive gland of slugs and its influence in biomarkers following transplantation between a relatively unpolluted and a chronically metal-polluted site

Changes in cell-type composition (CCTC) is a general phenomenon that takes place in the digestive gland epithelium of stressed molluscs. The aim of the present work was to determine whether CCTC is a reversible process in the digestive gland of sentinel slugs chronically exposed to metal pollution and how CCTC affects metal accumulation parameters and different cell and tissue biomarkers of exposure and effect. Slugs (Arion ater) from an abandoned zinc mine were transferred to a relatively unpolluted site and the other way around for 3, 10 and 28 d. The volume density of black silver deposits (Vv_BSD) after autometallography, and metallothionein (MT) levels were used as biomarkers of exposure to metals and CCTC and lysosomal responses were selected as effect biomarkers. Results indicated that slugs were sensitive to recent metal pollution; however, slugs chronically exposed to metals presented some characteristic features and were less responsive to pollution cessation without signs of CCTC reversal.     

Effects of tributyltin on the energy metabolism of pen shell (Atrina pectinata japonica)

We examined the effects of tributyltin (TBT) on aerobic and anaerobic energy metabolism of pen shell (Atrina pectinata japonica). We exposed pen shells to TBT at nominal concentrations of 0 (control) and 1.0microg/l for 72h under aerobic condition. At the end of the exposure, half of the pen shells in each treatment were wrapped in plastic wrap to simulate exposure to hypoxia and held at 25 degrees C for another 12h. The concentrations of the products of energy metabolism, namely lactate, pyruvate, fumarate and succinate, in adductor muscle were measured. The exposure to TBT under aerobic condition significantly elevated lactate, pyruvate and fumarate concentrations (p<0.001). After subsequent exposure to anaerobic condition, the mean concentration of succinate in the TBT treatment group was 64% of that in the control group, but there were no significant differences. Our results suggest that the energy metabolism of pen shell is disrupted by exposure to TBT.     

Tributyltin exposure causes brain damage in Sebastiscus marmoratus

Tributyltin (TBT) is a ubiquitous marine environmental contaminant characterized primarily by its reproductive toxicity. However, the neurotoxic effect of TBT has not been extensively described, especially in fishes which have a high number of species in the marine environment. This study was conducted to investigate the neurotoxic effects of TBT at environmental levels (1, 10, and 100ngl(-1)) on female Sebastiscus marmoratus. The results showed that TBT exposure induced apoptosis in brain cells of three regions including the pallial areas of the telencephalon, the granular layer of the optic tectum, and the cerebellum. In addition, the increase of reactive oxygen species and nitric oxide levels, and the decrease of Na(+)/K(+)-ATPase activity were found in the brain. The results strongly indicated neurotoxicity of TBT to fishes. According to the regions in which apoptosis was found in the brain, TBT exposure might influence the schooling, sensory and motorial functions of fishes.     

Tributyltin distribution in the coastal environment of Peninsular Malaysia

The occurrence of tributyltin (TBT) is reported in the coastal waters of a few selected sites in Peninsular Malaysia. Water, bivalves and sediment samples collected were analysed specifically for TBT using sensitive analytical methods which involved a solvent extraction procedure with appropriate clean-up followed by graphite furnace atomic absorption spectrometric measurements. The levels of TBT in the seawater in unexposed areas were found in the range from <3.4 to 20 ng litre(-1) as compared to coastal areas with high boat and ship activities where TBT levels in seawater were generally above 30 ng litre(-1), with the highest level found at 281.8 ng litre(-1). TBT levels in the tissues of random cockle and soft-shell clam samples from local markets were found in the range from <0.5 to 3.7 ng g(-1) wet weight. The levels of TBT found in green mussel samples both from the market (23.5 ng g(-1) wet weight) and those from a mussel farm (14.2 ng g(-1) wet weight) indicate slight accumulation of TBT. In sediments, TBT levels were found ranging from <0.7 ng g(-1) dry weight in unexposed coastal sites to as high as 216.5 ng g(-1) dry weight for a site within a port area.