Oxidative stress, lysosomal damage and dysfunctional autophagy in molluscan hepatopancreas (digestive gland) induced by chemical contaminants.

Autophagy is a highly conserved evolutionary survival or defence process that enables cells and organisms to survive periods of environmental stress by breaking down cellular organelles and macromolecules in autolysosomes to provide a supply of nutrients for cell maintenance. However, autophagy is also a part of normal cellular physiology that facilitates the turnover of cellular constituents under normal conditions: it can be readily augmented by mild environmental stress; but becomes dysfunctional with severe oxidative stress leading to cellular pathology. The molluscan hepatopancreas or digestive gland provides a versatile and environmentally relevant model to investigate lysosomal autophagy and stress-induced dysfunctional autophagy. This latter process has been implicated in many animal and human disease conditions, including degenerative and neurodegenerative diseases, as well as obesity related conditions. Many environmental pollutants have also been found to induce dysfunctional autophagy in molluscan hepatopancreatic digestive cells, and in this study, the marine blue mussel Mytilus galloprovincialis was exposed for 7 days to: 0.1 µM, 1 µM and 10 µM concentrations of fluoranthene and phenanthrene (PAHs); chlorpyrifos and malathion (organophosphorus compounds); atrazine (triazine herbicide); copper (transition metal) and dodecylbenzene sulphonic acid (LAS, surfactant). The marine snail or periwinkle, Littorina littorea, was also exposed to phenanthrene, chlorpyrifos and copper. Indices of oxidative stress, cell injury and dysfunctional autophagy were measured (i.e., lysosomal membrane stability, protein carbonyls, lipofuscin, and lysosomal accumulation of lipid or lipidosis). Evidence of oxidative stress, based on the elevation of lipofuscin and protein carbonyls, was found for all compounds tested; with chlorpyrifos being the most toxic to both species. Dysfunctional autophagy was induced by all of the compounds tested in both species, except for atrazine in mussels. This failure of normal autophagy was consistently associated with oxidative stress. Autophagic dysfunction is an important emerging feature in the aetiology of many disease conditions in animals and humans; and an explanatory conceptual mechanistic model has been developed for dysregulation of autophagy in response to oxidative stress.

Integration of biochemical, histochemical and toxicogenomic indices for the assessment of health status of mussels from the Tamar Estuary, U.K.

The aim of this study was to examine whether a combination of biochemical, histopathological and toxicogenomic data could be used as a valuable tool for the assessment of biological risk associated with pollutants within the Tamar River and Estuary, S.W. England, U.K. Accordingly, biochemical and histopathological biomarkers (protein carbonyls, lipofuscin, neutral lipids, lysosomal stability [N-acetyl-ß-hexosaminidase and neutral red], lysosomal volume, ferric reducing antioxidant power [FRAP] and malonaldehyde [MDA]) and gene expression profiles were assessed in 5 sites from the Tamar River and Estuary (Neal Point, Town Quay, Wilcove, Cremyll Ferry and Whitsand; and a reference site, Trebarwith Strand, N. Cornwall). PAHs were measured in mussel tissue and sediment and metals were measured in mussel tissue only. Data from the biomarkers was integrated into a Mussel Expert System (MES) model to produce a simple assessment of mussel stress. Clear gradients of mussel toxicity were identified by the biomarkers (with the exception of neutral lipids) with the highest impacted animals found furthest up the Tamar, whilst the MES was unable to identify a gradient of effect. Gene expression profiles also indicated a gradient of stress with the greatest number of significantly up- or down- regulated genes found at the uppermost 2 sites. The MES did, however, determine that mussels from all sites, except the reference site, were highly stressed; a conclusion that could not be inferred from the biomarker data alone. It is concluded that the MES is a valuable tool that permits integration and interpretation of complex sets of biomarker data by identifying the biological meaning of biomarker changes.

Improved removal of estrogenic and pharmaceutical compounds in sewage effluent by full scale granular activated carbon: impact on receiving river water.

Sewage effluents are widely recognised as the main source of emerging contaminants, such as endocrine disrupting chemicals (EDCs) and pharmaceuticals in surface waters. A full-scale granular activated carbon (GAC) plant has been installed as an advanced technology for the removal of these contaminants, in a major sewage treatment works (STW) in South-West England as part of the UK National Demonstration Programme for EDCs. This study presented for the first time, an assessment of the impact of a recently commissioned, post-tertiary GAC plant in the removal of emerging contaminants in a working STW. Through regular sampling followed by solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS), a significant reduction in the concentrations of steroidal estrogens was observed (>43-64%). In addition, significant reductions were observed for many of the pharmaceutical compounds such as mebeverine (84-99%), although the reduction was less dramatic for some of the more widely used pharmaceuticals analysed, including carbamazepine and propranolol (17-23%).

Dissolved nutrients in the Tweed Estuary, UK: inputs, distributions and effects of residence time.

Results are presented from a campaign in which dissolved inorganic nutrients were measured in the Tweed Estuary, UK. The data utilised here were derived from surveys undertaken on a monthly basis from January to August 1997. There was consistency between the nutrient concentrations observed in the freshwater reaches of the tidal estuary and earlier measurements made several km upstream, in the freshwater river. Typically, nitrate was of the order of 100 microM and silicate approximately half this, whereas phosphate and nitrite were typically approximately 1 microM. Seasonal fluctuations in freshwater nitrate and silicate were very large, with concentrations exceeding (respectively) 300 and 100 microM in winter and less than 30 and 10 microM in summer. The estuarine nutrients surveyed followed this general seasonal pattern. A simple box model was used to investigate the temporal behaviour of a hypothetical estuarine nutrient. The model demonstrated that large deviations from the 'broad-brush' seasonal pattern for nitrate and silicate could be expected to occur within the estuary as a result of runoff and residence-time variations. The model also demonstrated that mid-estuarine peaks observed in the longitudinal distributions of phosphate could only have arisen as a result of non-freshwater, wastewater phosphate inputs to the estuary. Without this mid-estuarine input, rapid flushing rates would have ensured that transients due to changing freshwater phosphate concentrations, runoff and tides (via the residence time) would have had relatively little impact on the classic, linear 'conservative' mixing diagram for the estuary. Conservative mixing was largely applicable to the other nutrients.

Seasonal variability of salinity, temperature, turbidity and suspended chlorophyll in the Tweed Estuary.

Results are presented from a campaign of measurements that were undertaken to examine seasonal variability in physical and chemical fluxes and processes within the Tweed Estuary during the period September 1996-August 1997. The study utilised monthly surveys, each of approximately 1 week duration. This article interprets a subset of the salinity, temperature, turbidity [suspended particulate matter (SPM) levels] and chlorophyll a data. Measurements discussed here were obtained throughout the estuary during high-speed transects that covered the region between the tidal river and the coastal zone. Longitudinal distributions of surface salinity depended strongly on freshwater runoff. During high runoff the surface salinity was low and the freshwater-saltwater interface (FSI) was located close to the mouth. The reverse was true at times of low runoff. Salinity stratification was generally strong. During the surveys, river runoff temperatures ranged from approximately 2 to 18 degrees C and coastal waters (approximately 33 salinity) from approximately 6 to 15 degrees C. Turbidity was low throughout the campaign (SPM < 30 mg l(-1)). Because of rapid flushing times (one or two tides), turbidity tended to mix conservatively between river and coastal waters. Higher coastal turbidity was associated with stronger wind events, and higher fluvial turbidity with spate events. Suspended chlorophyll a levels were usually low throughout the estuary (typically < 2 microg l(-1)) and showed large spatial variability. Because of the rapid flushing of the estuary, it is hypothesised that it was not possible for several algal cell divisions to occur before algae were flushed to the coastal zone. A 'bloom' occurred during the May 1997 survey, when chlorophyll a levels reached 14 microg l(-1). Higher chlorophyll a concentrations at that time occurred at very low salinities, indicating that these waters and algae were largely fluvially derived, and may have resulted from increasing springtime solar irradiation.

Concentrations of suspended particulate organic carbon in the tidal Yorkshire Ouse River and Humber Estuary.

Data are presented for particulate organic carbon (POC) and particulate nitrogen (PN) concentrations in the Humber Estuary and tidal River Ouse Estuary. The POC data were derived from approximately monthly surveys and are consistent with data reported for suspended particulate matter (SPM) in the non-tidal River Ouse (the freshwater river) and with SPM, or bed sediments, in estuarine ecosystems such as the Mississippi, Delaware, San Francisco Bay, Tolo Harbour, the Vellar Estuary and Cochin Backwater, as well as the Loire, Gironde, Ems and Tamar Estuaries. Relative to the dry weight of SPM, the Humber-averaged organic carbon and nitrogen percentages during the year February 1995-March 1996 were 2.6 +/- 0.6% (mean and S.D.) and 0.21 +/- 0.04%, respectively. The ratio of Humber-averaged POC to Humber-averaged PN was 13 +/- 3. Higher POC levels were observed near the Humber's mouth and in the adjacent coastal zone during 'bloom' conditions, and in the upper estuarine reaches during large, winter and springtime freshwater inflows. At these times of high runoff, the POC content of SPM increased progressively up-estuary from the coastal zone to the tidal River Ouse. When inflows became very low, during late spring to early autumn of 1995, both the freshwater-saltwater interface (FSI) and the strengthening turbidity maximum (TM) moved further up-estuary and the POC content of SPM in the upper reaches of the Ouse became lower compared with that immediately down-estuary. This led to a poorly defined POC maximum near the confluence of the Humber, Ouse and Trent, before POC eventually decreased again towards the coastal zone. The lower POC contents in the upper estuarine reaches of the tidal Ouse may have been partly due to POC respiration by heterotrophic bacteria attached to SPM within the TM, consistent with the severe oxygen depletion observed there during high turbidity, summertime spring tides.