Okadaic acid induces morphological changes, apoptosis and cell cycle alterations in different human cell types

Okadaic acid (OA) is a marine toxin produced by dinoflagellate species which is frequently accumulated in molluscs usual in the human diet. The exact action mechanism of OA has not been described yet and the results of most reported studies are often conflicting. The aim of this work was to evaluate the OA effects on morphology, cell cycle and apoptosis induction by means of light microscopy and flow cytometry, in three different types of human cells (leukocytes, HepG2 cells and SHSY5Y cells). Cells were treated with a range of OA concentrations in the presence and absence of S9 fraction. OA induced morphological changes in all the cell types studied, and cell cycle disruption only in leukocytes and neuronal cells. SHSY5Y cells were the most sensitive to OA assault. Results obtained in the presence and absence of metabolic activation were similar, suggesting that OA acts both directly and indirectly. Furthermore, OA was found to increase the subG(1) region in the flow cytometry cell cycle analysis, suggesting induction of apoptosis. These results were confirmed by the employment of specific methodologies for studying apoptosis such as caspase 3 activation and annexin V staining. Increases in the apoptosis rate were obtained in all the cells treated in the absence of S9 fraction, accompanied by increases in caspase 3 activation, suggesting that apoptosis induced by OA is a caspase 3-dependent process. Nevertheless, in the presence of S9 fraction no apoptosis was detected, indicating a metabolic detoxifying activity, although necrosis was observed in neuroblastoma cells.     

Assessing reliable indicators to sewage pollution in coastal soft-bottom communities

Physicochemical characteristics of sediment and benthic communities were studied in the proximity of seven sewage outfalls with differences in flow and wastewater treatment in the Western Mediterranean Sea. Redox potential was the only abiotic parameter which showed a pattern related with distance to outfalls, whereas granulometry, percentage of organic matter, metal concentrations or pH did not show changes related with outfall presence. Benthic community analysis proved to be the most suitable monitoring tool. The results showed that the highest impacted stations corresponded with those closest to outfall with the highest flow and only pre-treatment, whilst a decrease of this tendency was detected in the locations where secondary treatment takes place. Meta-analysis showed a decrease of amphipods and tanaids abundance as well as redox potential, as the indicators with the clearest response to sewage presence.     

Epiphyte toxicity bioassay for ecotoxicological and coastal monitoring

Marine epibionts are organisms that grow on submerged surfaces. Those found on seagrass leaves are especially important because of their interactions with the plants, their contribution to primary production in these ecosystems, and their role as food source for heterotrophic fauna. Given the relative lack of ecotoxicological studies on epibionts, the aim of this study is to evaluate the effect of environmental pollution on epiphytes experimentally attached to artificial devices (mimes) consisting of thermally-sealed silicone tubes supported on bamboo sticks that mimic the morphology of seagrasses and serve as an anchor surface for marine epibionts. Mimes were installed on the sea floor in subtidal waters of the Rio San Pedro (Cádiz), collected after 28 days, and incubated in the laboratory with environmental concentrations of atrazine (herbicide), Irgarol (anti-fouling substance), and copper. Tube-dwelling diatoms formed the major component of the epiphyte community. Average surface covering, chlorophyll, and biomass content did not show significant differences between controls and treatments. The glutathione peroxidase activity increased significantly with 4 μg L^?1 of atrazine and 5 μg L^?1 of copper. This enzymatic activity increase seems to be sufficient to prevent oxidative cellular damage by removing reactive oxygen substances produced by oxidative stress; in addition to this enzyme, there might be other antioxidant enzymes which have not been measured in this study, that have also protected the organism from oxidative damage. Thus, the measurement of antioxidant enzymatic activity in epiphytes may be a useful toxicity indicator for coastal biomonitoring.     

Essential (Cu) and nonessential (Cd and Pb) metals in ichthyofauna from the coasts of Sinaloa state (SE Gulf of California)

With the aim of giving an overview on concentration and distribution of Cd, Cu, and Pb in fish from the coasts of Sinaloa state (SE Gulf of California), specimens with different feeding habits were collected in five locations. Sampling occurred between June 2003 and March 2004. Metal analyses on fish tissues were made by graphite furnace (Cd, Pb) and flame (Cu) atomic absorption spectrophotometry. Metal concentrations in tissues of carnivorous fish were grouped together and compared with corresponding concentrations in non-carnivorous fish; Cu and Pb levels were significantly (p?<?0.05) higher in liver of non-carnivorous species. Though no samples exceeded the maximum level set in international legislation for fish, from the perspective of the public health and considering the legal limits of fishery products for human consumption, Cu concentrations were exceeded (in tissues different from muscle) in four carnivorous and five non-carnivorous species according to the Australian legislation. In the case of Cd, two carnivorous species (Pomadasys leuciscus and Caulolatilus princeps) and one non-carnivorous species (Mugil cephalus), showed concentrations over the maximum level of 2 μg g^???1 dry weight considered in the Mexican legislation. Considering average amounts of fish consumption in Mexico, daily mineral intake (DMI) values for Cu and percentage weekly intake (PWI) of Cd and Pb were estimated; none of the analyzed metals in edible portion of analyzed fish could be detrimental to humans.     

Multivariate analyses of the effect of an urban wastewater treatment plant on spatial and temporal variation of water quality and nutrient distribution of a tropical mid-order river

Freshwater resources are increasingly scarce due to human activities, and the understanding of water quality variations at different spatial and temporal scales is necessary for adequate management. Here, we analyze the hypotheses that (1) the presence of a wastewater treatment plant (WWTP) and (2) a polluted tributary that drains downstream from the WWTP change the spatial patterns of physicochemical variables (pH, turbidity, dissolved oxygen, and electrical conductivity) and nutrient concentrations (reactive soluble phosphorus, total phosphorus, nitrogen series, total nitrogen, and total dissolved carbon) along a mid-order river in SE Brazil and that these effects depend on rainfall regime. Six study sites were sampled along almost 4 years to evaluate the impacts of human activities, including sites upstream (1–3) and downstream (5–6) from the WWTP. The impacts were observed presenting an increasing trend from the source (site 1) towards Água Quente stream (site 4, the polluted tributary), with signs of attenuation at site 5 (downstream from both WWTP and site 4) and the river mouth (site 6). Input of nutrients by rural and urban runoff was observed mainly at sites 2 and 3, respectively. At sites 4 and 5, the inputs of both untreated and treated wastewaters increased nutrient concentrations and changed physicochemical variables, with significant impacts to Monjolinho River. Seasonal variations in the measured values were also observed, in agreement with the pluviometric indexes of the region. Univariate analyses suggested no effect of the WWTP for most variables, with continued impacts at sites downstream, but non-parametric multivariate analysis indicated that these sites were recovering to chemical characteristics similar to upstream sites, apparently due to autodepuration. Therefore, multivariate methods that allow rigorous tests of multifactor hypotheses can greatly contribute to determine effects of both point and non-point sources in river systems, thus contributing to freshwater monitoring and management.     

Pond bank access as an approach for managing toxic cyanobacteria in beef cattle pasture drinking water ponds

Forty-one livestock drinking water ponds in Alabama beef cattle pastures during were surveyed during the late summer to generally understand water quality patterns in these important water resources. Since livestock drinking water ponds are prone to excess nutrients that typically lead to eutrophication, which can promote blooms of toxigenic phytoplankton such as cyanobacteria, we also assessed the threat of exposure to the hepatotoxin, microcystin. Eighty percent of the ponds studied contained measurable microcystin, while three of these ponds had concentrations above human drinking water thresholds set by the US Environmental Protection Agency (i.e., 0.3 μg/L). Water quality patterns in the livestock drinking water ponds contrasted sharply with patterns typically observed for temperate freshwater lakes and reservoirs. Namely, we found several non-linear relationships between phytoplankton abundance (measured as chlorophyll) and nutrients or total suspended solids. Livestock had direct access to all the study ponds. Consequently, the proportion of inorganic suspended solids (e.g., sediment) increased with higher concentrations of total suspended solids, which underlies these patterns. Unimodal relationships were also observed between microcystin and phytoplankton abundance or nutrients. Euglenoids were abundant in the four ponds with chlorophyll concentrations >?250 μg/L (and dominated three of these ponds), which could explain why ponds with high chlorophyll concentrations would have low microcystin concentrations. Based on observations made during sampling events and available water quality data, livestock-mediated bioturbation is causing elevated total suspended solids that lead to reduced phytoplankton abundance and microcystin despite high concentrations of nutrients, such as phosphorus and nitrogen. Thus, livestock could be used to manage algal blooms, including toxic secondary metabolites, in their drinking water ponds by allowing them to walk in the ponds to increase turbidity.     

Comparison of two metal surveys by moss Tortula ruralis in Budapest, Hungary

Moss transplants of Tortula ruralis were used as active biomonitoring organisms as part of a monitoring study to assess the metals (Al, Cd, Cu, Cr, Fe, Ni, Pb, V and Zn) associated with ambient particles on mosses in Budapest, the capital town of Hungary. The moss samples were collected in a nature conservation area from a semi-arid sand grassland (Festucetum vaginatae danubiale), less than 1 month before transplantation. Moss cushions were exposed to pollution in Budapest during October–November 1993 and February–March 1994. In the study area, 16 sites were marked out as measuring sites, in accordance with the structure of the city. A similar stand was established in the Botanical Gardens of the Szent István University in Gödöll? as a control site. ICP-AES analysis of moss for metals showed the ability of Tortula ruralis to accumulate the metals under study. Control site showed lower impact in comparison to the other sites.