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PubMedDate not supplied

Recovery of polluted ecosystems: the case for long-term studies.

Recovery of marine ecosystems from pollution has tended to receive less attention than the study of new or continuing impacts, but such studies are important in charting recovery from acute incidents and following legislation to deal with chronic contamination. Recovery is inevitably a long-term process, and where such studies have been made they are often too short-lived. Interest quickly wanes following an acute incident and governmental bodies rapidly switch to new legislative priorities for chronic inputs. We review three case studies: recovery of dogwhelk populations after local extinction by tributyl tin leachates from anti-fouling paints; recovery of rocky shore communities from oil spills; and recovery of estuarine ecosystems from industrial and urban development. We then make some generalisations about recovery processes before making a plea for long-term studies of polluted areas.

Animals↗
PubMedDate not supplied

Mechanisms of imposex induction in the mud snail, Ilyanassa obsoleta: TBT as a neurotoxin and aromatase inhibitor.

The occurrence of imposex, imposition of male sex characteristics on female snails, has been extensively documented throughout the world. Tributyltin (TBT) and other organotins have been causally linked to imposex induction at levels as low as 2 ng/l. There are several proposed mechanisms of action. First, TBT has been shown to be neurotoxic and to accumulate in snail ganglia. Peptide hormones control sexual differentiation in gastropods, and one hypothesis is that TBT acts as a neurotoxin to abnormally release the peptide hormone Penis Morphogenic Factor (PMF). However, PMF has not been characterized to date. The neuropeptide APGWamide significantly induces imposex in the mud snail, Ilyanassa obsoleta, at 10(-16) moles sub-cutaneous (SQ) injection over 2 weeks, and could be the PMF in this species. A second hypothesis is that TBT inhibits aromatase activity leading to increased testosterone levels and decreased estradiol. In vitro studies with snail digestive gland microsomes showed that TBT-dosed snails not exhibiting imposex had a 52% reduction in aromatase activity. Although the role of vertebrate sex steroids is not known in gastropods, it is possible that the combination of changes in peptide and steroid hormones may lead to imposex induction at extremely low doses of TBT.

Animals↗
PubMedDate not supplied

Dumpton syndrome reduces the tributyltin (TBT) sterilising effect on Nucella lapillus (L.) by limiting the development of the imposed vas deferens.

TBT induces imposex and sterilisation in female Nucella lapillus. Interestingly, there are populations where individuals (both males and imposexed females) may suffer from a genetic aberration [Dumpton Syndrome (DS)] which results in the development of no penis (i.e. aphally). This anomaly confers a so far unexplained protection against TBT. Six populations with contrasting DS frequency were sampled; the imposex stage in females and the mean calibre of every vas deferens (VDC) were assessed. While the sex ratio in the 600 specimens examined was close to 1, aphally was more frequent in females than in males (31 and 7%, respectively). VDC was significantly higher in normal than in aphallic (DS) specimens, independently of sex and/or site. It is concluded that the vas deferens imposed by TBT onto DS females is less prone to develop thick enough so as to interfere with normal egg laying.

Animals↗
PubMedDate not supplied

Vancouver workshop: overview and synthesis.

Investigators from the six PICES member nations conducted a May, 1999 interdisciplinary research assessment of Vancouver Harbour, BC, Canada. Sediment contaminants were analyzed at several sites along an assumed pollution gradient extending eastward into the harbour from the relatively less-impacted Howe Sound area (located outside and seaward, to the west). Subsets of a few representative benthic biota (English sole for general contaminants; specific mollusks for TBT) at the sampling sites were concurrently assessed for specific tissue contaminants. Measurements of biotic response to contaminants-across a wide spectrum of biological and ecosystem organization, and ranging from selected biomarkers and tissue responses, to observations at the organism and community level-were likewise conducted. Parameters examined included: P-450 expression, EROD induction and histopathologic assessment of toxicopathic liver lesions in English sole; prevalence and frequency of imposex in neogastropods; perturbations in benthic community responses to pollutants; tissue assays of various bivalve shellfish species for PSP. Results affirmed a consistent east-to-west trend in declining contaminant-associated effects, across a range of biological assessments, thus providing strong evidence that these effects are pollution-related. Specific findings revealed a significant distortion of benthic community structure along the pollution gradient, as well as biomarker evidence of histological and biochemical stress in English sole from central and inner harbour sites. Although the study reaffirms prior assumptions that various species of neogastropods have been extirpated from the harbour, most likely from the antifoulant uses of tributyltin, some of the adverse impacts from TBT may be declining. Also noted were strong biochemical inferences that sediments in the relatively "uncontaminated" Howe Sound, may well contain significant P450-inducing pollutants which were not included as analytes in the design of this limited and preliminary assessment. During the brief sampling period, PSP contamination did not appear to be a significant issue for the harbour.

Animals↗
PubMed2003

Imposex as a biomonitoring tool for marine pollution by tributyltin: some further observations.

Imposex, i.e., the imposition of male sexual characteristics on female neograstropods, has been used worldwide as a bioindicator of pollution by the antifouling agent TBT as well as to assess the related ecological impact. The recent total ban on the use of TBT in the maritime industry has been partly based on the use of such a biomarker. This is mainly based on the assumption that no other pollutant, or environmental stress, is able to induce such a biological response. Nonetheless, several authors had challenged this idea. The present paper will present further field observations on imposex in Hexaplex trunculus from Malta (Central Mediterranean) which may help clarify this issue. Furthermore, the incidence of imposex in this species will be related to body burdens and to the environmental levels of organotins as measured analytically. First histological observations will also be presented which may help clarify the ecological significance of this response in this species. Finally a number of alternative scenarios regarding the relationship between imposex and TBT will be discussed.

Animals↗
PubMed2001

Effects of endocrine modulating substances on reproduction in the hermaphroditic snail Lymnaea stagnalis L.

Various man-made agents like pesticides, industrial chemicals and some natural substances have the potential to alter hormonal pathways that regulate reproductive processes in certain species of wildlife. Until now, only a few investigations have been undertaken to determine the effects of these substances on reproductive capacities (fecundity and fertility) in exposed invertebrate aquatic species. In this study one of the most abundant mollusc of European limnic systems, the hermaphroditic gastropod Lymnaea stagnalis was investigated to determine the effects of endocrine modulating substances on reproductive parameters. Known endocrine modulating substances were tested using the following nominal concentrations; Tributyltin (TBT in ng Sn/l) and beta-sitosterol at 1, 10 and 100 ng/l, respectively, and 4-nonylphenol (4-NP) at 1, 10 and 100 microg/l. In addition, experiments were carried out with 1, 10 and 100 ng/l of t-methyltestosterone. All the testing was carried out on recently matured adults of Lymnaea. Fifteen to twenty snails per treatment were exposed for between 7 and 12 weeks in a semi-static test with a weekly exchange of testwater. Shell height and weight and mortality of the adults, egg production, hatching rate of the eggs, and histopathology of the adult snails were analysed. The same parameters were investigated on F(1) generation snails from exposed parents at two dates (1 week after exposure and at the end of the exposure duration). Treatments with TBT and 4-NP had only slight effects on the egg production of the adults and hatching rate of the eggs. However, increased histopathological changes were observed in epithelial tissues of the adult snails, e.g. lung and foot also characterised by extreme inflammatory processes. While beta-sitosterol and t-methyltestosterone had no effect on the shell height and weight or the mortality of adult snails nor on the egg production or ensuing egg hatching rate, beta-sitosterol treated snails presented a distinct atrophy of the albumen gland and so did t-methyltestosterone, albeit with a lower degree of atrophy. The observed histopathological effects due to exposure to tributyltin or 4-NP are suggested to lead to long-term adverse reproductive effects mediated by an impairment of the fitness of the snails. In the experiments the steroid-dependant (beta-sitosterol and t-methyltestosterone) degeneration of the albumen gland caused no obvious adverse effects on the fecundity nor fertility of the adults or on F(1)-generation. However, the impact on fertility following a prolonged exposure to high concentrations of the phytoestrogen cannot be predicted.

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PubMed2001

Induction of apoptosis in the blue mussel Mytilus galloprovincialis by tri-n-butyltin chloride.

Induction of apoptosis by tri-n-butyltin (TBT) in gill tissue of the mussel Mytilus galloprovincialis was investigated. The terminal dUTP nick-end labeling technique (TUNEL) was used to detect cells displaying DNA fragmentation within gill structures. Genomic DNA fragmentation was detected as characteristically ladder-like pattern of DNA fragments induced by single injection of different doses of TBT (1-5 microg/g) below the mantle, directly into the pallial fluid, after 24 h of incubation. DNA degradation of higher order DNA structure, as well as reduced G(0)/G(1) cell cycle region (the sub-G(1) region) was detectable after 1.5 h of TBT incubation. Presence of apoptotic cells in mussels' gills was indicated by the selective loss of G(2)/M cells concomitant with the appearance of cells with decreased DNA content in S and G(0)/G(1) cell cycle regions. The effect of the TBT on cell cycle in a mussel gill was a dose related and exposure time depending. The possible mechanism of induction of apoptosis in vivo in gill tissue of mussel treated with TBT is discussed.

Animals↗
PubMed2001

Effects of tributyltin and other metals on the phenoloxidase activating system of the tunicate, Styela plicata.

Toxic metals, such as tributyltin (TBT), contribute substantially to anthropogenic pollution in many estuarine environments. Animals that live in those environments, particularly invertebrate filter feeders like tunicates, are likely to be exposed to substantial metal contamination. This study investigates the effects of TBT and other metals on the phenoloxidase activity of the estuarine tunicate, Styela plicata, in an effort to identify a biochemical marker of metal pollution. Hemocytes harvested from S. plicata that were exposed to tributyltin or copper in aquaria had significantly enhanced phenoloxidase activities relative to non-exposed controls. This enhanced phenoloxidase activity could be explained by an increased frequency of morula cells, which contain high levels of phenoloxidase's proenzyme, prophenoloxidase. Unlike those from tunicates exposed to metals in aquaria, the phenoloxidase activities of hemocytes incubated with tributyltin in vitro were significantly reduced when compared with hemocytes cultured without tributyltin. The ability of tributyltin to decrease phenoloxidase activity in tissue culture may reflect its known inhibitory effects on calcium-dependent signaling systems such as those involved in the exocytosis of prophenoloxidase from morula cells.

Animals↗
PubMed2001

Sediment-associated tri-n-butyltin chloride and its effects on osmoregulation of freshwater-adapted 0-group European flounder, Platichthys flesus (L.).

The disruption of osmoregulatory processes was examined in European flounders exposed to environmental concentrations (150 ng TBTCl g(-1) dry weight sediment) of sediment-associated tri-n-butyltin chloride (TBTCl), by using radiotracers to measure changes in hydromineral fluxes and water balance. The water permeabilities of TBTCl-exposed fish varied during the course of the experiment and were significantly lower than those of the corresponding controls that did not change significantly with time. It was found that the maximum decrease in water permeability of TBTCl-exposed fish occurred after 14 days; thereafter there was an increase towards control values. However, there was a differential reduction of the diffusional (P(d)) and osmotic (P(os)) permeability coefficients, where the former decreased more rapidly than the latter, reflecting the reduction of diffusional membrane permeability and the increasing importance of osmotic permeability. In fish exposed to TBTCl sodium efflux and drinking rates were significantly increased but Na(+)/K(+)-ATPase activities and urine production rates were not affected. The effects of TBTCl exposure are also manifested at the level of the whole organism by a reduction in the increase of the body length of exposed fish, when compared to controls. It was concluded that tributyltin-n-chloride in sediments is capable of significantly disrupting the osmoregulatory functions of a benthic estuarine fish, at concentrations found in the sediments of Southampton Water and the River Itchen.

Animals↗
PubMed2002

Genotoxic, cytotoxic and ontogenetic effects of tri-n-butyltin on the marine worm, Platynereis dumerilii (Polychaeta: Nereidae).

The genotoxic, cytotoxic and ontogenetic (embryo-larval) or developmental effects of tri-n-butyltin (TBT), were investigated in Platynereis dumerilii. Following the determination of maximum tolerated dose with regard to ontogenetic effects and mortality, early life stages of P. dumerilii were exposed to a range of TBT concentrations. Subsequently, the embryo-larvae were analysed for evidence of genotoxicity and cytotoxicity. Genotoxicity was assessed using cytogenetic endpoints that included the frequency of sister chromatid exchanges and chromosomal aberrations from metaphase spreads. Cytotoxicity was evaluated by determining the proliferative rate index of the growing embryo-larval cells using 5-bromodeoxyuridine labelling of the chromosomes or fluorescence plus Giemsa staining technique. TBT-exposed embryo-larvae of P. dumerilii exhibited sensitivity similar to that of other invertebrates, indicating that P. dumerilii is a suitable ecotoxicity test species. The results also suggested dose-dependent effects for genotoxic and cytotoxic end points in relation to TBT exposure. The present study highlights the need to elucidate the relative importance of direct genotoxic and indirect effects through production of genotoxic hormonal derivatives.

Animals↗
PubMed2003

Early life exposure to environmental levels of the aromatase inhibitor tributyltin causes masculinisation and irreversible sperm damage in zebrafish (Danio rerio).

To determine whether early life exposure to tributyltin (TBT), an aromatase inhibitor, impaired reproductive function in fish, Danio rerio were exposed to environmentally realistic levels (0.01-100 ng l(-1)) of TBT from 0 to 30, 30 to 60, and 0 to 70 days post-hatch, and the sex ratio and sperm motility of the adults examined 3-5 months after cessation of exposure. Fish exposed for 70 days to 0.1 ng l(-1) of TBT, a concentration presently below the detection limit in water, showed a male biased population which produced a high incidence of sperm lacking flagella. At 1 ng l(-1), the motility of sperm was significantly lower than that of control fish, while at 10 ng l(-1), all sperm lacked flagella and, at 100 ng l(-1), milt volume had increased. The effect of exposure on sex ratio was similar after exposure from 0 to 70 and 0 to 30 days, but even 100 ng l(-1) gave only 65% males after exposure from 30 to 60 days. Effects on sperm motility and morphology and on milt volume were less pronounced after 30 day than 70 day exposure. Our data suggest that screening for aromatase inhibiting activity and assessment of its risks in early life to human and wildlife fertility needs to be urgently addressed, and that the reproductive toxicity of TBT may presently be underestimated.

Animals↗
PubMed1999

Tributyltin triggers apoptosis in trout hepatocytes: the role of Ca2+, protein kinase C and proteases.

The purpose of the present study was to study the mechanisms involved in the induction of apoptosis and by tributyltin (TBT) in rainbow trout hepatocytes, and to examine the role of intracellular Ca2+, protein kinase C (PKC) and proteases in the apoptotic process. The intracellular Ca2+ chelator BAPTA-AM has a suppressive effect on TBT-mediated apoptosis. However, exposure to the ionophore A23187 is not sufficient to induce apoptosis in trout hepatocytes. The results obtained also show that TBT stimulates PKC gamma and delta translocation from cytosol to the plasma membrane in trout hepatocytes after 30 min of exposure. However, PKC gamma translocation is down-regulated after 90 min of treatment. The addition of protein kinase inhibitors (staurosporine and H-7) not only fails to inhibit apoptosis induced by TBT, but also leads to enhancement of DNA fragmentation. These inhibitors also afford a remarkable protection against the loss of plasma membrane integrity caused by TBT exposure. PMA, a direct activator of PKC, fails to stimulate DNA fragmentation. In addition, Z-VAD.FMK is an extremely potent inhibitor of TBT-induced apoptosis in trout hepatocytes, indicating that the activation of ICE-like proteases is a key event in this process. The cysteine protease inhibitor N-ethylmaleimide also prevented TBT-induced DNA fragmentation. Taken together, these data allow for the first time to suggest a mechanistic model of TBT-induced apoptosis. We propose that TBT could trigger apoptosis through a step involving Ca2+ efflux from the endoplasmic reticulum or other intracellular pools and by mechanisms involving cysteine proteases, such as calpains, as well as the phosphorylation status of apoptotic proteins such as Bcl-2 homologues.

Animals↗
PubMed2000

Cytotoxicity of tributyltin in rat hippocampal slice cultures.

The neurotoxic effects of tributyltin (TBT), an endocrine-disrupting chemical, were evaluated in organotypic slice cultures of immature rat hippocampus. Confocal microscopy study with propidium iodide showed that TBT induced severe neuronal death in a concentration- and time-dependent manner with CA3 > CA1 > dentate gyrus ranking of vulnerability of the hippocampal subfields. Dead or damaged neurons exhibited chromatin condensation, which is one of the morphological characteristics of apoptosis, as revealed by acridine orange staining. TBT neurotoxicity was alleviated by application of free radical scavengers or antioxidants, such as catalase, superoxide dismutase, Trolox and alpha-tocopherol but not by ascorbic acid or N-acetyl-L-cysteine, which suggests an involvement of free radicals, particularly reactive oxygen species. Neurons displayed a long-lasting increase in intracellular Ca2+ concentrations after TBT treatment. Although neither N-methyl-D-aspartate (NMDA) receptor inhibitors nor voltage-sensitive Ca2+ channel blockers protected hippocampal neurons against TBT neurotoxicity, non-NMDA receptor antagonist completely prevented TBT-induced neurodegeneration. These data suggest that TBT provokes apoptosis-like neuronal cell death, which might be mediated by intracellular Ca2+ elevation and free radical generation via non-NMDA receptor activation.

Acridine Orange↗
PubMed2000

The neuropeptide APGWamide induces imposex in the mud snail, Ilyanassa obsoleta.

We investigated whether neuropeptides which control sexual differentiation in mollusks can induce imposex-a condition where female snails grow male accessory sex organs after exposure to tributyltin (TBT). Mud snails, Ilyanassa obsoleta, were dosed with one of four neuropeptides: APGWamide, conopressin, LSSFVRIamide, or FMRFamide for seven or fourteen days. TBT and testosterone (T) were used as positive controls and induced imposex as expected. APGWamide significantly induced imposex, with a threshold dose near 10(-)(16) moles. The other neuropeptides had no effect on imposex induction. We propose that TBT could act as a neurotoxin to induce imposex via abnormal release of APGWamide.

Animals↗
PubMed1983

The pattern of myelin proteins in triethyltin (TET) intoxication.

Myelin proteins isolated from the brain of Wistar rats intoxicated chronically with triethyltin sulfate (TET) according to the technique of ETO et al. (1971) were investigated. Among the various protein fractions, the Agraval protein happened to be most evidently affected, demonstrating considerably reduced percentages. However the results did not prove that any of the individual myelin proteins was specifically affected by TET intake. The interesting point in chronic TET poisoning was that some clinical symptoms as well as disturbances in myelin proteins demonstrated a clear tendency to retrogression despite of the continued intoxication. These observations indicate obviously that during chronic TET intake, some kind of biochemical adaptation to the noxious action of the poison on the myelin sheath takes place.

Animals↗
PubMed2002

Development of imposex and accumulation of butyltin in the tropical muricid Thais distinguenda transplanted to a TBT contaminated site.

The muricid Thais distinguenda (ca. 1,300 individuals) was collected from a pristine island in the Andaman Sea and tagged prior to translocation to an area of intense shipping activities. A minimum of 50 tagged individuals were recaptured 1, 2, 3, 5, 6, 7, 8 and 12 months after transplantation. The animals were separated into small (< or = 25 mm) and big (>25 mm) specimens based on total shell length and both incidence of imposex and body burden of butyltin (BT) were determined. After 3 months 16.7% of the small size and none of the big size class had developed imposex. After 5 months the incidence of imposex in the big and small size classes increased steadily with time reaching 86.4 and 80%, respectively after 1 year. Time after transplantation was significantly correlated with the incidence of imposex for the big size class (Spearman Rs=1; P=0.008); RPLI (Spearman Rs=1; P=0.008) and VDSI (Spearman Rs=1; P=0.008); and the small size class: incidence of imposex (Spearman Rs=1; P=0.006); RPLI (Spearman Rs=1; P=0.006) and VDSI (Spearman Rs=1: P=0.007). The development of imposex was not significantly related to body size after five months (chi square P<0.001). The body burden of tributyltin was not significantly different between the two size classes (paired t-test ) but the smaller animals had significantly higher concentrations of both dibutyltin (DBT; P<0.005) and monobutyltin (MBT; P<0.001). The use of transplantation experiments in addition to imposex surveys and analysis of body burden of tributyltin (TBT) can enhance the interpretation of the results and understanding of the time scale involved with the development of imposex.

Animals↗
PubMed2003

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.

Animals↗
PubMed2002

Inhibitory effects of tributyltin on dopamine biosynthesis in rat PC12 cells.

The inhibitory effects of tributyltin chloride (TBTC) on dopamine biosynthesis in PC12 cells were investigated. Twenty-four hour exposure to TBTC at 0.5 microM showed 32.9% inhibition of dopamine content: the IC(50) value of TBTC was 0.72 microM. Dopamine content decreased at 6 h and reached a minimal level at 24 h after exposure to TBTC at 0.5 microM. The decreased dopamine level was maintained for up to 48 h. Under these conditions, tyrosine hydroxylase (TH) activity was inhibited at 6 h following the treatment with TBTC and the activity was maintained at a reduced level for up to 48 h (20-35% inhibition at 0.5 microM of TBTC). TH mRNA level also started to decrease at about 6 h and reached a minimal level at 24 h after exposure of PC12 cells to TBTC. In addition, treatment with L-3,4-dihydroxyphenylalanine (L-DOPA) at 20-50 microM increased the intracellular dopamine content in PC12 cells and the increase of dopamine level by L-DOPA was significantly inhibited after exposure to TBTC at 0.5-2.0 microM for 24 h. These results indicate that TBTC decreases dopamine content by the inhibition of TH activity and TH mRNA level in PC12 cells. It is, therefore, proposed that TBTC may exacerbate the Parkinson's symptoms because of the inhibition of dopamine biosynthesis and dopamine increase induced by L-DOPA.

Animals↗