Stress-70 proteins in marine mussel Mytilus galloprovincialis as biomarkers of environmental pollution: a field study

In the present work we have investigated levels of stress-70 proteins in the gills of mussel Mytilus galloprovincialis collected seasonally from subtidal rocky shores at 6 different sites of the Rovinj coastal area (Northern Adriatic, Croatia). 1-D analysis (SDS-PAGE) using monoclonal mouse antibodies anti-HSP70 detected two bands of stress-70 proteins, 70 and 72 kDa constitutively present during the year. 2-D analysis (IEF+SDS-PAGE) proved that the antibodies used detected HSP70 (pI 5.7-5.9) and HSP72 (pI 5.5-5.6). The quantification of stress-70 proteins was possible using 200 ng of external HSP70 protein standard included on every blot. Maximal levels of HSP72 and HSP70 were observed in mussels in summer (September), and minimal levels in winter (December), and only HSP70 showed significant correlation with the sea temperature (r=+0.822, p<0.05). Acclimatization of mussels to a different lower salinity under experimental conditions proved that small changes in sea salinity (Delta=2 psu) could not cause significant stress-70 proteins induction. Results indicated that there are significant differences in HSP70 and HSP72 content in mussels from the control site (S-1) and mussels from other sampling sites with urban and industrial pollution. The usefulness of stress-70 proteins as biomarkers of environmental pollution is discussed.     

Stress-70 as indicator of heavy metals accumulation in blue mussel Mytilus edulis

The induction of stress-70 (Hsp70) in the blue mussels Mytilus edulis from southern Baltic after exposure to cadmium, lead and copper, administrated individually and/or as a mixture, was investigated by means of standard Western blotting using a clone 3A3 monoclonal antibody. The general course of the stress-70 induction was found to be similar for all tested metals. It could be characterised by three main features: (1) The induction of stress-70 proteins was only slightly increased at low metal concentration. (2) Strong induction of stress-70 occurred at high concentration of metals. Combination of metals (Cd+Cu) was found to increase the stress-70 level more intensely than related concentrations of singularly applied metals. (3) Five weeks of long depuration had no effect on stress-70 levels. Experiments revealed different induction of stress-70 as a response to cadmium, lead and copper exposure and confirm that the applied technique might serve as a versatile tool to assess invertebrate stress reaction to environmentally relevant heavy metals.     

Organochlorine compounds and heavy metals in the soft tissue of the mussel Mytilus galloprovincialis collected from Lake Faro (Sicily, Italy)

Three hundred samples of Mytilus galloprovincialis were collected from five stations (north, south, east, west and centre) of Lake Faro to evaluate the concentrations of organochlorine compounds and heavy metals. Quantitative determinations of organochlorine pesticides and PCBs were made by GC-ECD and confirmed with GC-MS. Concentrations of "essential" (Cu, Se and Zn) and "toxic" (As, Cd, Hg and Pb) metals were determined by an atomic absorption spectrophotometer. The results obtained show the low residue levels of p,p'-DDE in six samples of M. galloprovincialis from southern (7.00-11.00 ng/g w.w. and 148.3-275 ng/g l.w.) and western (7.60-15.37 ng/g w.w. and 126.7-256.2 ng/g l.w.) areas of Lake Faro. No appreciable residues of PCBs were found in any of the samples examined. Zn concentrations (range 11.0-18.5 microg/g w.w.) were higher than Cu (range 188.3-396.0 ng/g w.w.) and Se (range 93.5-288.9 ng/g w.w.) in all areas of origin. Cd (range 41.9-63.8 ng/g w.w.), Pb (range 64.8-93.0 ng/g w.w.) and Hg levels (range 5.7-13.1 ng/g w.w.) showed lower concentrations than permitted MRLs. The As levels were below detection limits for the all mussel samples. In conclusion, the absence of PCBs, the low levels of p,p'-DDE, the concentrations of Cd, Hg and Pb below permitted MRLs in M. galloprovincialis, used as a "biological indicator", show that Lake Faro is not at contamination risk from these contaminants and moreover is free from health problems for the consumer of mussel products.     

Monitoring of the impact of Prestige oil spill on Mytilus galloprovincialis from Galician coast

In November 2002, the Prestige oil tanker was wrecked in front of Galician coast (NW of Spain), spilling near 63,000 tons of heavy oil until February 2003. Contamination produced was very extensive (70% of Galician beaches were reached by the oil) but heterogeneous, alternating intensely affected zones with neighbour locations where the repercussion was minimal. The objective of this study was to monitor sea environment contamination caused by Prestige oil spill during an 11-month period (August 2003-June 2004, nine samplings) in two locations of Galician coast with different geographical properties (Lira and Ancoradoiro beaches), by means of chemical determination of total polycyclic aromatic hydrocarbons (TPAH) in seawater, and using as exposure biomarker TPAH content in mussel (Mytilus galloprovincialis) tissues, and as effect biomarker DNA damage in mussel gills, evaluated by the comet assay. In addition, recovery ability of the mussels was determined after a 7-day stay in the laboratory. TPAH contents in seawater were very high in the earliest samplings, but then they maintained below 200 ng L(-)(1), similar to reference seawater. However, TPAH levels in mussel tissues were more variable: they increased again from January 2004, probably due to the adverse meteorological conditions that turned over the sea bottom and dispersed the oil accumulated in sediments. In most samplings, these levels decreased during the recovery stage. DNA damage in oil-exposed mussels was significantly higher than in reference mussels, both before and after the recovery phase, but they did not differ to one another. Comet tail length was slightly reduced during the recovery stage, indicative of a certain DNA repair in exposed mussels. This study showed up the importance of monitoring sea contamination events during an extended time, not only in evaluating the presence of the contaminants in the environment but also in determining their bioaccumulation and their effect on the exposed organisms.     

Metal and metallothionein level in the heat-treated cytosol of gills of transplanted mussels Mytilus galloprovincialis Lmk

A stock of mussels Mytilus galloprovincialis was transplanted over 1 year to four sites in a semi-enclosed bay in Croatia which is under the influence of various sources of pollution. The positive correlation of metal (Cd, Zn, Cu, Mn, Fe) and metallothionein (MT) tissue contents based on the analysis of heat-treated cytosol of gills with shell mass, as an indicator of mussel age, indicated to accumulation of metals, as well as the increase of MTs with mussel age. The principal component analysis (PCA) revealed that 74% of total variance of obtained results could be explained through two principal components. The first principal component was highly correlated with MT, Cd and Zn indices (metal or MT content/shell mass), and the second one with Cu, Mn, and Fe indices, as well as the gill index. High correlation of MTs with Zn and Cd is consistent with their affinity for binding to MTs.     

Experimental studies on the biokinetics of 134Cs and 24Am in mussels (Mytilus galloprovincialis).

The biokinetics of 134Cs and 241Am in mussel species contaminated through water pathway has been studied under laboratory conditions. At equilibrium, the concentration factors for 134Cs and 241Am in small and large mussels were 2.80 and 2.57 and 200 and 150, respectively. The concentration factor of 134Cs in soft parts of the mussels was significantly high than whole body and shell tissue. However, the concentration factors of 241Am in soft parts and shell tissue samples were found to have similar rates. The depuration kinetics of the radionuclides were described by two-component exponential models. The biological half-lives at slow components between small and large mussels did not change significant, and were found to be 46.8-46.5 and 72.2-75.3 days for 134Cs and 241Am. The depuration kinetics of 134Cs and 241Am in soft parts described a single-component exponential model and the biological half-lives were found to be 29.4 and 41.1 days, respectively.     

Lysosomal responses and metallothionein induction in the blue mussel Mytilus edulis from the south-west coast of Iceland

It has recently been emphasized that high levels of inorganic and organic micropollutants (particularly organometals, POPs and PAHs) may be present in coastal waters at high latitudes, stressing the need to evaluate the effects of contaminants on marine organisms from sub-arctic zones. With this aim, specimens of the blue mussel Mytilus edulis were sampled in polluted and reference areas along the south-west coast of Iceland in July 2004. Samples were collected from the intertidal zone at three sites in Reykjavik harbour which are differently exposed to contaminants, and at three reference coastal sites, two located along the Reykjanes Peninsula and the third one on the northern part of Hvalfjordur fiord. Lipofuscin content, neutral lipid accumulation and lysosomal enlargement were evaluated in digestive cells from cryostat sections of the mussel hepatopancreas, and quantified by automated image analysis. Metallothionein induction was also determined in the same tissue. Results indicate that mussels from the inner part of Reykjavik harbour, which is the most sheltered and most influenced by extensive shipping traffic, were the worst affected, with the highest values in neutral lipids, lipofuscin and lysosomal swelling. At the other two harbour sites, mussels exhibited lower values, similar to those observed in organisms collected in Hvalfjordur fiord and in bay of Osar. Mussels from Kuagerdi had the lowest values.     

Glutathione S-tranferases and cytochrome P450 activities in Mytilus galloprovincialis from the South coast of Portugal: effect of abiotic factors

To assess the health of aquatic organisms, biomarkers that measure changes at the biochemical level have been used as effective early warning tools in ecological risk assessment. In order to develop an integrated risk assessment strategy for the south coast of Portugal, mussels Mytilus galloprovincialis were collected from six sites along the coast with different organic contaminant characteristics. Additionally, an active biomonitoring approach was followed by transplanting indigenous mussels from site 4 to 6 and vice versa (from site 6 to 4) for 28 days. PAHs and PCBs contents were measured and the associated responses of phase I and phase II detoxification mechanisms evaluated by measuring cytochrome P450 and GST activities. GST activity was also determined on different tissues (gills, digestive gland, foot, mantle and gonads) of M. galloprovincialis and the impact of abiotic parameters (temperature, salinity, pH, conductivity and dissolved oxygen) on the GST activity assessed. Results indicate that CYP 450 follow the same pattern of PAHs and a direct relationship exist between CYP 450 and PAH levels from the different sites. Although there is a decrease between GST and PAHs concentrations it was not significant. The majority of the GST activity was in the gills and the digestive gland (around of 75% of the activity measured in all tissues) followed in decreasing order by the mantle, gonads. An inverse relationship between GST activity and salinity was detected along with temperature although not significant. These two biomarkers respond to changes of these two groups of compounds and to salinity especially for GST. In conclusion CYP 450 in mussels gives a reliable response as biomarker for organic contaminants in risk assessment in the South Coast of Portugal.     

Immediate biomarker responses to benzo[a]pyrene in polluted and unpolluted populations of the blue mussel (Mytilus edulis L.) at high-latitudes

Immediate biomarker responses of two high-latitude populations of the blue mussel Mytilus edulis to benzo[a]pyrene (B[a]P) were evaluated. Mussels collected from a clean and a polluted site in southwest Iceland were exposed to the nominal dose of 100 microg B[a]P L(-l) for 3 h, after 4 days of acclimatization in clean seawater. To test the sensitivity to the toxicant and immediate biological responses, the following biomarkers were used: DNA single strand breaks, heart rate and feeding rate. All the biomarkers revealed differences between the study sites. Irrespective of the origin of the organisms, the short time exposure to the high B[a]P concentration did not induce DNA single strand breaks or significantly affect the feeding rate. However, the heart rate results showed significantly different responses. The mussels from the polluted site (Reykjavík harbour) increased their heart rate when exposed to B[a]P, while no difference was observed between the heart rate values of the individuals from the clean site (Hvassahraun). The mussels seem to sense the pollutant they have been previously exposed to, and their acute response indicates physiological adaptation to the polluted environment. The results indicate limited sensitivity and temporal predictivity, i.e. transient measurable changes of these biomarkers, as well as showing that the background of the organisms should be considered when evaluating short-term biomarker responses to contaminants.     

Cellular alterations in Mytilus galloprovincialis (LMK) and Tapes philippinarum (Adams and Reeve, 1850) as biomarkers of environmental stress: field studies in the Lagoon of Venice (Italy)

A long-term biomonitoring study was carried out in the Lagoon of Venice (North-East Italy) with the aim of evaluating variations in biological responses to environmental stress in estuarine bivalves. Two different species, the mussel Mytilus galloprovincialis and the clam Tapes philippinarum, both widespread in the Lagoon, were studied in several sites. Two cellular biomarkers: lysosomal membrane stability in digestive cells and thickness of digestive epithelia, were evaluated in native organisms (on a seasonal basis), and in organisms which have been transferred from a reference site to several differently influenced ones. Results indicate that, to some extent, both test and organisms were able to highlight site-specific differences, but the effects of pollution were generally more easily detected by reduction in lysosomal stability than by reduction in digestive tubule epithelium. Further findings show that the inherent variability of a number of natural parameters, particularly in the reference sites, produced less effective results when biological responses in the reference organisms were compared with the polluted ones. The assessment of the two conditions was most valuable when they referred to the 25-75% range of values comprised within the seasonal medians, recorded respectively in control and polluted sites. Impaired from steady states were most effectively distinguished when the control values were medians from two reference locations. Lately, the overall results indicate that both biomarkers are more suitably deployed through the translocation approach, revealing it to be more sensitive than traditional biomonitoring, at least in the sense that it may overcome problems related to the adaptation of native organisms to sub lethal chronic pollution levels.     

Study on the behavior of the heavy metals Cu, Cr, Ni, Zn, Fe, Mn and 137Cs in an estuarine ecosystem using Mytilus galloprovincialis as a bioindicator species: the case of Thermaikos gulf, Greece

Mussels are worldwide recognized as pollution bioindicators and used in Mussel Watch programs, because they accumulate pollutants in their tissues at elevated levels in relation to pollutant biological availability in the marine environment. The present study deals with the use of Mytilus galloprovincialis as a local bioindicator of heavy metal and (137)Cs contamination in an estuarine ecosystem (Thermaikos gulf, Greece in Eastern Mediterranean). M. galloprovincialis samples were collected monthly from two aquaculture farms during the period April to October 2000. Analyses for the heavy metals Cu, Cr, Ni, Zn, Fe, Mn and (137)Cs showed that the concentrations measured were low and similar to those from other non-polluted Mediterranean areas. In terms of the two sampling stations, there were no statistically significant differences between them. On the contrary, the seasonal evolution of either heavy metals or (137)Cs levels presented high variation. The levels were found to increase during the cold period of the year, especially for Cu, Zn, Mn and Cr which are essential for life. Stable metals were positively inter-related and moreover, metals more involved in biochemical activities seem to present more correlations than others with less significant role in the metabolism of the organisms.     

Modeling of breakpoint reaction in drinking water distribution pipes

A mathematical model accounting for concurrent mass transfer and a series of chemical reactions under the breakpoint chlorination is developed to predict disinfectant concentration profiles in the drinking water distribution pipe. The model is validated by comparing its numerical solutions to experimental data in the literature. The impact of important parameters on the model performance is examined by a sensitivity analysis. Practical applications of the model to minimize water quality deterioration in the distribution system are discussed in view of the results. This work can provide insight into the factors that influence all of the fundamental reactions and disinfectant transport of the breakpoint reaction. Operational criteria for the chlorination of distributed water are derived.     

Health risks from large-scale water pollution: trends in Central Asia

Limited data on the pollution status of spatially extensive water systems constrain health-risk assessments at basin-scales. Using a recipient measurement approach in a terminal water body, we show that agricultural and industrial pollutants in groundwater-surface water systems of the Aral Sea Drainage Basin (covering the main part of Central Asia) yield cumulative health hazards above guideline values in downstream surface waters, due to high concentrations of copper, arsenic, nitrite, and to certain extent dichlorodiphenyltrichloroethane (DDT). Considering these high-impact contaminants, we furthermore perform trend analyses of their upstream spatial-temporal distribution, investigating dominant large-scale spreading mechanisms. The ratio between parent DDT and its degradation products showed that discharges into or depositions onto surface waters are likely to be recent or ongoing. In river water, copper concentrations peak during the spring season, after thawing and snow melt. High spatial variability of arsenic concentrations in river water could reflect its local presence in the top soil of nearby agricultural fields. Overall, groundwaters were associated with much higher health risks than surface waters. Health risks can therefore increase considerably, if the downstream population must switch to groundwater-based drinking water supplies during surface water shortage. Arid regions are generally vulnerable to this problem due to ongoing irrigation expansion and climate changes.     

Changes in the taxonomic structure of herpetobiont arachnids along the gradient of pollution with emissions from a copper smelter

Changes in the dynamic density of herpetobiont arachnids and the ratio of spider life forms have been studied in southern taiga forests transformed under the impact of emissions from a copper smelter. Along the pollution gradient, the dynamic density of spiders decreases; web-spinning spiders are replaced by ambush spiders, since the abundance of families Lycosidae and Gnaphosidae increases while that of the family Linyphiidae decreases; and spatial variation in the abundance of arachnids increases. One harvestmen family (Nemastomatidae) completely disappears near the copper smelter.     

Lead content in roadside vegetation as indicators of atmospheric pollution

Leaves of six plant species from 31 different locations in Ibadan, Nigeria, were analysed for their lead content. The lead concentrations found ranged between 12 and 32 μg/g, 50 m or more away from the roadside, between 47 and 115 μg/g in the residential low traffic density areas, 5 m from the roadside, and between 165 and 312 μg/g in the commercial high traffic density roadsides, 5 m away from the roadside.     

MRIO model-based study on water nitrogen pollution transfer embodied in international trade

This study uses the global pollutant emission databases and global input–output model in 2015 to calculate the impact of international trade on global water nitrogen emission patterns, based on considering the total amount of pollutant transfer and pollutant emission intensity of trade flows The main conclusions are as follows: (1) There are always a large amount of water nitrogen emissions transferring from developed economies to developing economies embodied in their bilateral trade activities. Small amount of transfers are of some areas with similar endowments of agricultural resources or long distances. (2) In 2015, the net import of water nitrogen pollution embodied in China’s trade was 160,000 tons, accounting for 2.72% of the global water nitrogen imports. The sharp increases in cereal imports, together with high food storage as well as high pollution intensity embedded in trade are the main reason. It is recommended that through applying alleviations such as agricultural machinery assistance and technical training to accelerate the transfer and spread of agricultural technology in Africa, Asia, and other regions, thus helping increase agricultural production productivity in underdeveloped areas and reducing the pollution intensity embodied in trade flows from underdeveloped areas to developed areas.     

Dynamics of snowmelt water composition in conifer forests exposed to airborne industrial pollution

An analysis is made of the spatial variability of snowmelt water composition (within and between biogeocenoses), with regard to its long-term dynamics, in pine and spruce forests exposed to airborne industrial pollution from the Europe’s largest Severonikel Copper–Nickel Smelter Complex. Snowmelt waters from under the tree canopy, compared to those from intercrown areas, contain higher concentrations of chemical elements due to their washing and leaching from tree crowns. This is especially true of spruce forests, since the crowns of spruce trees have a high sorption capacity. Distinct trends in the long-term dynamics of snowmelt water composition, related to reduction of industrial emissions, are observed in background forest areas and defoliating forests but not in the vicinity of pollution sources. It is shown that the main factors determining these dynamics in forests of the Kola Peninsula are edificator tree species, airborne industrial pollution, and, possibly, an increase in the number of days with above-zero temperatures in the period of snow accumulation, which facilitates washing and leaching of chemical compounds from tree crowns.     

Trace metal levels in tree barks as indicators of atmospheric pollution

Barks of six species of trees were sampled at thirty-nine different locations in Ibadan, Nigeria and were analysed for the trace metals lead, cadmium, cobalt, copper, zinc, manganese, and iron. Lead levels in barks from areas with relatively high traffic density were generally higher than lead levels in barks from areas with low traffic density. The lead level recorded in the high traffic density areas, 40–140 μg/g, is low compared to values reported for barks of trees along busy roads in developed countries. For the other metals, no correlation with traffic density was observed.     

Trace element concentrations in tree rings as indicators of environmental pollution

Proton induced X-ray emission spectroscopy has been successfully used to measure variations in relative and absolute trace element concentration levels in tree rings. Valuable information on past concentration levels, stored in the tree rings have been obtained. Peaks associated with 15 elements have been identified in all X-ray spectra. Variations of trace elements levels in the tree rings reflect the variations in their availability from the environment.     

Input of heavy metals to the forest floor as a result of Cracow urban pollution

The influence of Cracow's urban pollution on small forest ecosystems was studied during 1998-2000. Seven sites located at different distances from the Cracow conurbation and from busy road were selected. Two sites were situated 26 and 31 km from Cracow town along an eastern transect, the other five-3, 8, 14, 26 and 35 km respectively, from Cracow-along a southern transect. At the beginning of September, five litter traps were set at each site. The quantity of litter fall, along with percentage composition of each species, were estimated for an area of 1 m(2). The concentration of Cd, Pb, Cu, Zn and Fe in the litter fall and the input of these metals to the forest floor were determined. The input of cadmium ranged from 84 microg m(-2) at Kornatka in 1998 to 382 microg m(-2) at Bonarka in 2000. The input of lead was about 10 times higher than cadmium and varied from 406 microg m(-2) at Weglówka in 1998 to 3717 microg m(-2) at Bonarka in 2000. Input of Cu, Zn and Fe to the forest floor showed the similar tendency. During 1998-2000, the highest heavy metal concentrations in the litter fall and their impact on the forest floor were always found at sites close to the Cracow conurbation and, in the eastern transect, close to a steel works. Input of metals to the forest floor tended to decrease with increasing distance from the centre of Cracow.