Organotin compounds (OTs) accumulate in fish easily, however, research on their influencing factors is still limited. This study collected 25 species of fish with different diets, habitats, and age from the Three Gorges Reservoir (TGR), the largest deep-water river channel-type reservoir in China, and analyzed the accumulation characteristics of OTs in these fish. The results showed that tributyltin (TBT) and triphenyltin (TPhT) were the dominant OTs in fish from the TGR. The correlation between OTs concentration and age, body length, and body weight varied with fish species. The concentrations of TBT and TPhT in carnivorous fish (mean, 25.78 and 11.69 ng Sn/g dw, respectively) were higher than those in other diet fish (P<0.01), but there was no significant difference in fish at different habitat water layers (P>0.05). In addition, the degradation rates of TBT and TPhT in different fish species were all below 50%. In summary, the accumulation of TBT and TPhT in fish is mainly influenced by diet, and both TBT and TPhT were difficult to degrade in fish. These results reveal the pollution characteristics of OTs in fish from the TGR, and can improve our understanding of the factors influencing TBT and TPhT accumulation in freshwater fish.

Protected areas (PAs) are dedicated and managed to achieve long-term conservation of nature with associated ecosystem services and cultural values. However, few studies have assessed contamination/pollution, one of the top five biodiversity threats, within these areas. In fact, more than 100,000 molecules used in consumer goods are potentially affecting over 250,000 protected areas distributed worldwide. As a result, chemical appraisal within PAs require specific approaches to be feasible. Herein, an approach for assessment of contamination in marine protected areas (MPAs) spread over large continental and/or global areas was proposed and tested using, as a case study, the impact of tributyltin (TBT) within Latin America. This approach included area delimitation, bibliographic survey, construction of attribute tables, data insertion in a Geographic Information System, overlapping with worldwide shapefiles of PAs, identification of affected PAs and evaluation of impacts using the outcomes against specific environmental quality guidelines/directives. A total of 259 records of TBT occurrence and/or its biological effect within 53 marine protected areas (including biosphere reserves, Ramsar sites and National parks) were gathered. Among affected species, were recorded 18 gastropods exhibiting imposex. In addition, 6 bivalves, 1 crustacean, and 3 fish species presenting TBT residues in tissues were seen in MPAs from Brazil, Argentina, Ecuador, Peru, Chile and Venezuela. Since levels of TBT recorded in several studied MPAs were enough do induce deleterious biological responses over marine biota, it was clear that conservancy goals of some \"protected\" areas are under threat. Considering that only TBT exposure was appraised, and these areas are likely to be concomitantly exposed to other classes of contaminants, the actual scenario can be even more frightening. Although initially applied as a TBT case study, the present approach enables an overview for other chemicals within protected areas worldwide.

Morphological malformations induced by tributyltin (TBT) exposure during embryonic development have already been characterized in various taxonomic groups, but, nonetheless, the molecular processes underlying these changes remain obscure. The present study provides the first genome-wide screening for differentially expressed genes that are linked to morphological alterations of gonadal tissue from chicken embryos after exposure to TBT. We applied a single injection of TBT (between 0.5 and 30 pg as Sn/g egg) into incubated fertile eggs to simulate maternal transfer of the endocrine disruptive compound. Methyltestosterone (MT) served as a positive control (30 pg/g egg). After 19 days of incubation, structural features of the gonads as well as genome-wide gene expression profiles were assessed simultaneously. TBT induced significant morphological and histological malformations of gonadal tissue from female embryos that show a virilization of the ovaries. This phenotypical virilization was mirrored by altered expression profiles of sex-dependent genes. Among these are several transcription and growth factors (e.g. FGF12, CTCF, NFIB), whose altered expression might serve as a set of markers for early identification of endocrine active chemicals that affect embryonic development by transcriptome profiling without the need of elaborate histological analyses.

Marine ecosystems are subject to anthropogenic change at global, regional and local scales. Global drivers interact with regional- and local-scale impacts of both a chronic and acute nature. Natural fluctuations and those driven by climate change need to be understood to diagnose local- and regional-scale impacts, and to inform assessments of recovery. Three case studies are used to illustrate the need for long-term studies: (i) separation of the influence of fishing pressure from climate change on bottom fish in the English Channel; (ii) recovery of rocky shore assemblages from the Torrey Canyon oil spill in the southwest of England; (iii) interaction of climate change and chronic Tributyltin pollution affecting recovery of rocky shore populations following the Torrey Canyon oil spill. We emphasize that \"baselines\" or \"reference states\" are better viewed as envelopes that are dependent on the time window of observation. Recommendations are made for adaptive management in a rapidly changing world.

Nuclear 9-cis retinoic acid receptors (retinoid X receptors, RXR) are promiscuous dimerization partners for a number of nuclear receptors. In the present study, we established a novel in vitro method for quantitative determination of the nuclear retinoid X receptors in rat liver. One type of high affinity and limited capacity RXR specific binding sites with the Ka value ranging from 1.011 to 1.727×10(9)l/mol and the Bmax value ranging from 0.346 to 0.567pmol/mg, was demonstrated. Maximal 9-cis retinoic acid (9cRA) specific binding to nuclear retinoid X receptors was achieved at 20°C, and the optimal incubation time for the 9cRA-RXR complex formation was 120min. From a number of endocrine disruptors, tributyltins and triphenyltins are known as RXR ligands. Our data confirmed the property of tributyltin chloride or triphenyltin chloride to bind to a high affinity and limited capacity RXR binding sites. Described optimal conditions for ligand binding to RXR molecules enabled us to calculate maximal binding capacity (Bmax) and affinity (Ka) values. This study provides an original RXR radioligand binding assay that can be employed for investigation of novel RXR ligands that comprise both drugs and endocrine disruptors.

Butyltin (TBT, DBT, and MBT) effects on molluscs, especially endocrine disruption in bivalves and gastropods, have been widely investigated. Imposex, the superimposition of male characters onto female gonochoristic Caenogastropods, is the most studied biological effect of TBT. TBT compounds are among the priority hazardous substances within Directives 2000/60/EC (WFD) and 2008/105/EC. The Environmental Quality Standards (EQSs) set by the WFD for TBT are quite difficult to quantify by means of chemical analysis, without the use of expensive and high performance methods. Assuming that EQSs set for TBT were derived from evidence of imposex development at very low concentrations, this specific biomarker could be used as an indirect measure of assessing levels of bioavailable BTs. Therefore, this study aims to validate the use of imposex development as an investigative tool to monitor the bioavailable fraction of BTs within the WFD, by comparing imposex levels and BT concentrations in Nassarius nitidus from the Venice Lagoon. BT concentrations and imposex levels in N. nitidus, collected in 2013, had decreased when compared to previous studies in the same area. Both VDSI and RPLI correlated positively with BT body burden in females, confirming that imposex is a valid tool to monitor bioavailable BTs. However, TBT is still a matter of concern in the Venice Lagoon, as TBT concentrations were still higher than its degradation products suggesting recent fresh TBT inputs in the studied area. To propose imposex levels as an indicator of the impact of BTs within the WFD, classification class boundaries and Ecological Quality Ratios were introduced. As a preliminary attempt, imposex levels were also compared to the OSPAR Commission EcoQOs which linked imposex levels in Nassarius reticulatus with TBT concentrations in water. Based on this comparison the degree of imposex development in the Venice Lagoon suggested that TBT concentrations in water should be over the EQS-AA concentration and, at one site, also over the EQS-MAC. From all the results obtained in this work, it appears that imposex evaluation in N. reticulatus could give information about the ecological status regarding BT compounds under the WFD and also be used to monitor their effects and support chemical analyses until more sensitive methods become available.

BACKGROUND: A variety of conditions (culture media, inocula, incubation temperatures) are employed in antifouling tests with marine bacteria. Shewanella algae was selected as model organism to evaluate the effect of these parameters on: bacterial growth, biofilm formation, the activity of model antifoulants, and the development and nanomechanical properties of the biofilms.The main objectives were: 1) To highlight and quantify the effect of these conditions on relevant parameters for antifouling studies: biofilm morphology, thickness, roughness, surface coverage, elasticity and adhesion forces. 2) To establish and characterise in detail a biofilm model with a relevant marine strain.
RESULTS: Both the medium and the temperature significantly influenced the total cell densities and biofilm biomasses in 24-hour cultures. Likewise, the IC50 of three antifouling standards (TBTO, tralopyril and zinc pyrithione) was significantly affected by the medium and the initial cell density. Four media (Marine Broth, MB; 2% NaCl Mueller-Hinton Broth, MH2; Luria Marine Broth, LMB; and Supplemented Artificial Seawater, SASW) were selected to explore their effect on the morphological and nanomechanical properties of 24-h biofilms. Two biofilm growth patterns were observed: a clear trend to vertical development, with varying thickness and surface coverage in MB, LMB and SASW, and a horizontal, relatively thin film in MH2. The Atomic Force Microscopy analysis showed the lowest Young modulii for MB (0.16 ± 0.10 MPa), followed by SASW (0.19 ± 0.09 MPa), LMB (0.22 ± 0.13 MPa) and MH2 (0.34 ± 0.16 MPa). Adhesion forces followed an inverted trend, being higher in MB (1.33 ± 0.38 nN) and lower in MH2 (0.73 ± 0.29 nN).
CONCLUSIONS: All the parameters significantly affected the ability of S. algae to grow and form biofilms, as well as the activity of antifouling molecules. A detailed study has been carried out in order to establish a biofilm model for further assays. The morphology and nanomechanics of S. algae biofilms were markedly influenced by the nutritional environments in which they were developed. As strategies for biofilm formation inhibition and biofilm detachment are of particular interest in antifouling research, the present findings also highlight the need for a careful selection of the assay conditions.

Micro-pollutant fluxes distribution throughout the physical separation and biological units of wastewater treatment plants (WWTPs) are very dependent ofsorption phenomena. The understanding and the control of the sorption stage is thus essential for the optimization of micro-pollutant removal in WWTPs, and particularly in biological treatments where these mechanisms influence the bioavailability towards micro-organisms. If the influence of the micro-pollutant physicochemical characteristics (e.g. Kow, pKa) on their ability to sorb on biological media (i.e. sludge) has been demonstrated, it appears that some other parameters, like the biosorbent characteristics, have to been taken into account. The aim of this study is thus to correlate the capacities of sorption of an environmentally relevant substance (tributyltin), with a thorough characterization of different types of sludge. The characterization of three biological media (raw, sonicated and flocculated activated sludges) is proposed according to various characterization parameters related to biochemical composition, aggregate size, rheological behaviour etc. The results show first that, whatever the sludge characteristics may be, the sorption mechanisms are very rapid and that an equilibrium state is reached after a few minutes. The influence of the sludge characteristics, notably the floc size and the chemical oxygen demand partition between solid and colloidal fraction, on sorption efficiency is demonstrated. A Langmuir modelling allows giving the maximum sorption capacity, as well as the binding energy for the three studied sludges, according to their physicochemical characteristics.

Discharge consents for tributyltin (TBT) from wastewater treatment works in the UK are set by the Environment Agency. These values are normally derived from the Environmental Quality Standards for concentrations of TBT in the receiving water, based on the volume of effluent discharged and subsequent dilution within the environment. In this study, a sampling program was designed to monitor concentrations of TBT (and other organotins) in the influent and effluent at each stage of treatment. Sampling was undertaken at 3 hourly intervals, over 24 hours, at 8 locations throughout the works for five days. Organotin concentrations were determined using capillary gas chromatography with flame photometric detection. The results demonstrated that TBT was consistently present in the influent at concentrations of approximately 0.1 microg l(-1). However, a \"pulse\" of TBT and monobutyltin (MBT) was observed on the third day when the concentration of TBT and MBT in the influent increased to 14 microg l(-1). This \"pulse\" of TBT and MBT was also observed as the wastewater passed through the unit treatment processes. Concentrations of both compounds were much reduced in the final effluent (TBT 2.5 microg l(-1) and MBT 6.9 microg l(-1)). Over the entire period, the average removal of TBT during primary treatment was 81%, and during secondary biological treatment was 71%. The overall removal efficiency of the plant, taking into account the recycling of returned liquors was 86% between influent and effluent for TBT. Removal efficiency of TBT was correlated to that of suspended solids, and it was demonstrated that on days when suspended solids removal was low, TBT removal was also reduced. The mass flow of organotins through the plant indicated that the majority of TBT was concentrated into the sewage sludge. Concentrations of TBT in sludge at the plant were approximately 18 microg kg(-1) (dry weight).

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.