Effect of metal accumulation on metallothionein level and condition of the periwinkle Littorina littorea along the Scheldt estuary (the Netherlands)

Metal (i.e. Ag, As, Ca, Cd, Co, Cu, Mn, Pb and Zn) and metallothionein (MT) concentrations in the soft tissue of Littorina littorea were measured along the heavily polluted Western Scheldt (WS) and relatively clean Eastern Scheldt (ES) estuary. Along the WS metal and MT levels in periwinkles reflected the known downstream decreasing pollution gradient. Surprisingly in ES animals As, Mn and Zn concentrations decreased from east to west reflecting past pollution. Compared to the WS metal concentrations of ES periwinkles were significantly lower and both estuaries were maximally discriminated from each other based on their Cd soft tissue concentration using a canonical discriminant analysis. Furthermore, no overall difference was found in MT levels among animals from both estuaries. Using previously obtained condition data (i.e. dry/wet weight ratio and lipid content) the relation between soft tissue metal concentration (i.e. Cd, Cu and Zn) and fitness indicators (i.e. MT and condition data) was examined using a canonical correlation analysis. Periwinkles with a high metal load (i.e. Cd and Zn) also had high MT levels but were in a relatively poor condition.     

Effects of heavy metal pollution on red wood ant (Formica s. str.) populations

We studied the species composition, mound population densities, relative abundance and colony sizes of red wood ants along a well known air pollution gradient of a copper smelter in Southwest Finland. The dominant species, Formica aquilonia, was further studied for heavy metal (Al, Cu, Cd, Ni, Zn, As, Pb, Hg) levels and morphological characters (body mass, head width, labial gland disease) of workers. We found five species belonging to Formica s. str., and two of them showed changes in their relative abundance, which could not be explained by natural habitat differences. Nest mound volumes were 34% smaller in the polluted area, suggesting that smaller colonies can be maintained there. The heavy metal levels in F. aquilonia workers were higher in the polluted area for all metals, except Hg. The largest relative differences between the study areas (polluted/unpolluted) were found for As (4.1), Ni (2.4), Cu (2.1) and Pb (1.8). Morphological characters of workers were not related to the heavy metal levels. Our data showed that red wood ants can tolerate relatively high amounts of heavy metals and maintain reproducing colonies even in a heavily polluted area, but on the basis of smaller colony sizes, pollution stress may also cause trade-offs in reproduction.     

Do wood mice (Apodemus sylvaticus L.) use food selection as a means to reduce heavy metal intake?

Food preference of wood mice from two with heavy metals polluted sites and two unpolluted sites was tested under laboratory and field conditions with two-way choice experiments. In the laboratory, wood mice preferred to eat acorns from unpolluted sites over acorns from polluted sites. Previous experience with polluted food had no influence on food choice. Preference was negatively related to acorn metal content. Furthermore, the nutrient content of the acorn endosperm was consistently lower in polluted sites. We therefore conclude that wood mice used absolute metal concentration in the acorn, nutrient content, or both as a food selection cue. The results of the laboratory experiment could not be confirmed under field conditions. We hypothesized that search time constraints due to the presence of predators, competitors and/or other stress factors in the field have prevented the mice to forage selectively.     

Relations between metals (Zn, Pb, Cd and Cu) and glutathione-dependent detoxifying enzymes in spiders from a heavy metal pollution gradient

We studied the relations between glutathione-dependent detoxifying enzymes and heavy metal burdens in the web-building spider Agelena labyrinthica (Agelenidae) and the wolf spider Pardosa lugubris (Lycosidae) from five meadow sites along a heavy metal pollution gradient. We assayed the activity of glutathione-S-transferase (GST) and glutathione peroxidases (GPOX, GSTPx), and glutathione (GSH) levels in both sexes. Except for GSH vs Pb content, we found significant correlations between GPOX and GSTPx activity and metal concentrations in females of A. labyrinthica. The highest activity of these enzymes measured in the web-building spiders was found in the individuals from the most polluted sites. In P. lugubris males significant correlations were found between GST and Pb and Zn concentrations, and between GPOX and GSTPx and the concentration of Cu. GST activity was higher in males collected from less polluted areas. Thus, detoxifying strategies against pollutants seemed to be sex-dependent. Actively hunting spiders had higher metal concentrations, maintaining lower activity of detoxifying enzymes and a lower glutathione level.     

Comparison of EDTA-enhanced phytoextraction and phytostabilisation strategies with Lolium perenne on a heavy metal contaminated soil

Phytoremediation is a promising and cost-effective strategy to manage heavy metal polluted sites. In this experiment, we compared simultaneously phytoextraction and phytostabilisation techniques on a Cd and Zn contaminated soil, through monitoring of plant accumulation and leaching. Lolium perenne plants were cultivated for 2 months under controlled environmental conditions in a 27.6 dm³-pot experiment allowing the collect of leachates. The heavy metal phytoextraction was promoted by adding Na-EDTA (0.5 g kg⁻¹ of soil) in watering solution. Phytostabilisation was assessed by mixing soil with steel shots (1%) before L. perenne sowing. Presence of plants exacerbated heavy metal leaching, by improving soil hydraulic conductivity. Use of EDTA for phytoextraction led to higher concentration of heavy metal in shoots. However, this higher heavy metal extraction was insufficient to satisfactory reduce the heavy metal content in soil, and led to important heavy metal leaching induced by EDTA. On the other hand, addition of steel shots efficiently decreased both Cd and Zn mobility, according to 0.01 M CaCl₂ extraction, and leaching. However, improvement of growth conditions by steel shots led to higher heavy metal mass in shoot tissues. Therefore, soil heavy metal mobility and plant metal uptake are not systematically positively correlated.     

Active biomonitoring of heavy metal pollution using Rosa rugosa plants

The purpose of this work was to evaluate the quality of a rural area near Faenza (Italy) by using an active biomonitoring approach, i.e., by placing homogeneous individuals of the perennial shrub Rosa rugosa in different sites throughout the area. Further sites, within the city or its environs, were used for comparison. Soil and leaves of R. rugosa were analyzed for their heavy metal content. The total heavy metal pattern of leaves closely paralleled the pattern registered in soil, with the highest content (both in total and assimilable forms) at the site in the urban area, which is exposed to heavy traffic. Pollen quality (abortiveness and viability) was also tested as a potential indicator of pollution. Pollen abortiveness was strictly related to Pb levels in leaves, while viability was inversely related to leaf Cr content. Our results suggest that R. rugosa has the potential to be a good biomonitoring system.     

Assessing the effects of heavy metal contamination on the proteome of the moss Pseudoscleropodium purum cross-transplanted between different areas

Protein expression was assessed in samples of Pseudoscleropodium purum cross-transplanted between one unpolluted (UNP) and two polluted (POLL) sites. Firstly, the level of expression (LE) of 17 proteins differed between native mosses from both types of sites, but differences were only maintained throughout the experiment for 5 of them. The LE of these five proteins changed over time in mosses transplanted from UNP to POLL and vice versa, becoming similar to that in autotransplants. However, these changes occurred slower than changes in the heavy metal concentrations measured in the same samples, and therefore they were not related to atmospheric pollution. Although the proteins identified were associated with moss metabolism, the expected growth reduction in samples autotransplanted within POLL (as a result of the down-regulation of photosynthesis-related proteins), did not occur. This supports the hypothesis that mosses growing in polluted areas adapt to heavy metal pollution and are able to reduce/overcome their toxic effects (i.e., reduced growth). Nevertheless, further specific research must be carried out to identify the proteins involved in this type of response, as lack of information on the bryophyte genome precludes us from reaching further conclusions.     

Temporal and spatial biomonitoring of heavy metals in Hong Kong coastal waters using Tetraclita squamosa

The barnacle Tetraclita squamosa (Crustacea: Cirripedia) was collected from 11 sites in the coastal waters of Hong Kong during three consecutive seasons, i.e. summer 1996, winter 1996-1997 and summer 1997. Barnacle soft tissues were analysed for cadmium, copper, manganese, iron and zinc using inductively coupled plasma atomic emission spectrophotometry. Analysis of covariance allowed inter-seasonal (temporal) and inter-site (spatial) comparisons of soft tissue metal concentrations to be made, from which bioavailabilities to T. squamosa have been inferred. Inter-seasonal variation was not greater than intra-seasonal and, therefore, seasonal variation in soft tissue metal concentrations can be considered relatively unimportant. Data were, however, characterised by high temporal variability. Spatial variation in metal bioavailabilities were consistent with earlier studies: cadmium soft tissue concentrations were elevated in barnacles collected from low salinity western waters, while copper and zinc concentrations were elevated in the urban-fringed central waters, in particular at one site, i.e. Aberdeen. In general, metal concentrations were much lower in barnacles collected from eastern water sites.     

A microcosm test of adaptation and species specific responses to polluted sediments applicable to indigenous chironomids (Diptera)

Chironomids may adapt to pollution stress but data are confined to species that can be reared in the laboratory. A microcosm approach was used to test for adaptation and species differences in heavy metal tolerance. In one experiment, microcosms containing different levels of contaminants were placed in polluted and reference locations. The response of Chironomus februarius to metal contaminants suggested local adaptation: relatively more flies emerged from clean sediment at the reference site and the reverse pattern occurred at the polluted site. However, maternal effects were not specifically ruled out. In another species, Kiefferulus intertinctus, there was no evidence for adaptation. In a second experiment, microcosms with different contaminant levels were placed at two polluted and two unpolluted sites. Species responded differently to contaminants, but there was no evidence for adaptation in the species where this could be tested. Adaptation to heavy metals may be uncommon and species specific, but more sensitive species need to be tested across a range of pollution levels. Factors influencing the likelihood of adaptation are briefly discussed.     

Response of kidney bean to concentration and chemical form of cadmium, zinc, and lead in polluted soils

Seven soils which had been polluted with heavy metals from a zinc smelter were sequentially extracted so that Cd, Zn, and Pb could be partitioned into five operationally defined geochemical fractions: exchangeable, carbonate, Fe-Mn oxide, organic, and residual fractions. Kidney beans were planted in the soils to examine the effect of concentration and chemical form of the metals in soil on the growth and metal uptake of the plants. The growth of kidney bean was restricted in heavy metal polluted soils compared with controls. Metal concentration and metal uptake by plants were correlated. The highest relationship was found between amount of metal uptake and the metal concentration in exchangeable + carbonate forms. The uptake of metals was according to their solubility sequence, i.e. Cd > Zn > Pb. The uptake rate of exchangeable + carbonate forms was the same for the three elements.     

Effects of heavy metal exposure on the condition and health of nestlings of the great tit (Parus major), a small songbird species

In this study we examined the possible effects of heavy metal exposure on the condition and health of great tit nestlings (Parus major) at four study sites along a pollution gradient near a large non-ferrous smelter in Belgium during three consecutive breeding seasons. Our results showed that nestlings were indeed exposed to large amounts of heavy metals. Excrements contained significantly higher concentrations of several heavy metals (silver, arsenic, cadmium, mercury, lead) near the pollution source than at study sites farther away. When taking into account the number of young in the nest at the time of sampling, nestling body mass and condition were significantly reduced at the most polluted site. Nestlings at the two most polluted sites fledged significantly later than at the least polluted site. We also observed growth abnormalities of the legs near the pollution source. Tarsus length, wing length and haematocrit values did not differ significantly among study sites.     

Metal accumulation in aquatic macrophytes from southeast Queensland,Australia

To determine the extent of metal accumulation in some aquatic macrophytes from contaminated urban streams in southeast Queensland, plants were sampled from six sites, along with contiguous sediments. In all, 15 different species were collected, the most common genera being Typha (Cattails or Bulrushes) and Persicaria (Knotweeds). Before heavy metal analysis, plants were further separated into various morphological tissues, and five selected samples were separated into various physiological tissues. The cadmium, copper, lead and zinc content of the plants were analysed using flames AAS. In general, plant roots exhibited higher metal concentrations than the contiguous sediments. Of the metals of interest, only for zinc was there a relatively clear pattern of increasing accumulation in aquatic macrophytes with increasing sediment metal concentrations. Comparison between morphological tissues of the sampled plants found that roots consistently presented higher metal concentrations than either the stems or leaves, however unlike previous studies, this investigation revealed no consistent trend of stems accumulating more metals than the leaves. For Typha spp., metal concentrations followed the order of roots > rhizomes > leaves, while for Persicaria spp. the order was roots > leaves > stems. The submerged species Myriophyllum aquaticum accumulated the highest levels of metals overall (e.g. Zn 4300 micrograms g-1 dry weight and Cd 6.5 micrograms g-1), and the emergent macrophytes also exhibited relatively high metal contents in their roots. The leaves of the submerged and floating-leafed species collected contained relatively high quantities of the four metals of interest, compared with the leaves of emergent aquatic macrophytes. In the Typha rhizome and Persicaria stem samples analysed for internal variation in metal content, there was a pattern of increasing metal concentrations towards the external sections of the stem, both for subterranean stems (rhizomes) and above-substrate stems. For Persicaria stems, no clear pattern was observed for cadmium and lead, the two metals investigated that are not required by plants for survival.     

Time to death response in carabid beetles exposed to multiple stressors along a gradient of heavy metal pollution

We investigated the responses of invertebrates inhabiting polluted environments to multiple stressors. Carabid beetles (Pterostichus oblongopunctatus F.) were subjected to food deprivation and insecticide treatment (dimethoate) to resolve trends associated with a gradient of heavy metal pollution. Metal concentrations along the gradient of five sites ranged from approximately 150 to 10,500 mg/kg Zn, 136 to 2600 mg/kg Pb, and 0.84 to 81.9 mg/kg Cd. There was no difference in body mass along the pollution gradient. However, the beetles originating from the most contaminated sites were significantly less tolerant to food deprivation than beetles from the reference site. Median survival time was 120 h for the two most polluted sites, compared with 168 h at the reference site. Beetles from the two most polluted sites were also significantly more susceptible to dimethoate at 0.1 microgram active ingredient/beetle. Median survival times were 12 and 123 h for beetles from the two most polluted sites and 359 h for the reference site. Carabid beetles exposed to chronic pollution, therefore, exhibit elevated susceptibility to additional stressors.     

Tissue, sex and age specific accumulation of heavy metals (Zn, Cu, Pb, Cd) by populations of the mole (Talpa europaea L.) in a central urban area

Heavy metal accumulation was studied in urban populations of moles, Talpa europaea L. (Insectivora) at three sites along the "Donaukanal" in Vienna. Kidneys, liver, lung, gonads, spleen, pancreas, femur, stomach + content, heart and skin were analysed for Zn, Cu, Pb and Cd contents. Female moles showed higher Zn concentrations in spleen, gonads and skin, higher Cd concentrations in femur and stomach and lower Cu content in kidneys compared to males. Age classes were determined using teeth section. A significant increase in Cd concentration with age was found in kidneys, liver, heart, lung, skin and femur, with the highest increase for kidney samples. Pb accumulated in bone tissue (femur) with increasing age. Although significant site differences in heavy metal concentrations of soil and vegetation samples existed, these differences were only partly reflected by tissue contents.     

The amphipod Hyalella azteca as a biomonitor in field deployment studies for metal mining

Specimens of the amphipod Hyalella azteca were deployed, in June-July 2003, along metal contamination gradients in two rivers affected by metal mining in the Abitibi - James Bay region, northwestern Québec. The amphipods were placed along with natural food items in small, acrylic cages and left in six riverine sites for 17 days. Twelve metals (As, Cu, La, Mn, Ni, Sb, Se, Tl, U, V, Zn, and CrO₄²⁻ modelled by WHAM VI) in transplanted H. azteca varied along metal contamination gradients in a consistent manner, i.e., as a function of metal exposure. Bioaccumulation of As, Cr, La, Ni, Sb, Se, Tl, U and V, as defined by a field BCF, was significantly correlated with their chronic toxicity potential towards the amphipod. We conclude that H. azteca may be a useful field biomonitor for metal mining. In addition, our results suggest that such biomonitoring programs should include less studied elements such as Se in mining effluents.     

Further investigation of the heavy metal content of the teeth of the bank vole as an exposure indicator of environmental pollution in Poland

The content of heavy metals in mineralised dental tissues is an indicator of the exposure of their mineral phase to heavy metals during the time of tooth development and dental tissue formation. Therefore, teeth have been used as bio-indicators which accurately reflect the environmental or dietary exposure of animals and humans to heavy metals. This study follows from the earlier work in which it was demonstrated that the teeth from bank voles inhabiting various environmentally polluted and non-polluted forests in Southern Poland were reliable indicators of exposure to heavy metals. Using analytical techniques employed in the earlier study heavy metal concentrations were obtained in the teeth of bank voles trapped in 1998-2000 to determine if efforts to clean up the environment could be detected in changes in heavy metal concentrations in the teeth. The results show that these efforts are reflected in lower concentrations of heavy metals in the teeth but that cross border contamination remains a problem.     

Lead reduces shell mass in juvenile garden snails (Helix aspersa)

In an earlier paper examining inherited tolerance to Pb, the shell growth of laboratory-bred offspring of Helix aspersa from contaminated sites was compared with that ofjuveniles from naive populations on dosed and undosed diets. Eight-week-old snails were fed either 500 microg g(-1) Pb or a control food in competitive trials between two populations. In the first series of trials, a parental history of exposure to Pb did not confer any advantage to either of two populations (BI and MI) competing with a na?ve population (LE). whether Pb was present in the diet or not. However, in the analysis of their metal concentrations reported here, LE are found to retain higher levels of Pb in the soft tissues than either BI or MI. Compared to their siblings on the unleaded diet, dosed LE and BI juveniles had lower soft tissue concentrations of Ca and Mg. Although the growth in shell height is unaffected by diet, LE and BI juveniles build lighter shells on the Pb-dosed diet, achieving around 75% of the shell mass of their controls. In contrast, the shell weights of dosed MI juveniles are depressed by only 15% and show no change in the essential metal concentrations of their soft tissues. A second experiment using five populations fed only the dosed food show that the shell weight/soft tissue weight ratios are comparable to the dosed snails of the previous experiment. Building a lighter shell thus appears to be the common response of all Helix populations to a high Pb diet, at least amongst juveniles. The reduction in its mass means that less Ca and Mg is added to the shell and, along with the lowered soft tissue concentrations observed in some populations. may be a consequence of an increased effort to excrete Pb. The possibility that the MI population shows a genotypic adaptation. perhaps as some form of modification of its Ca metabolism, is briefly discussed.     

Urban polluted forest soils induce elevated root peroxidase activity in Scots pine (Pinus sylvestris L.) seedlings.

Plant biomass. mycorrhizal status and root peroxidase activity were measured in ectomycorrhizal Scots pine (Pinus sylvestris L.) seedlings grown in urban polluted and native, non-polluted forest soils with added ammonium or potassium sulphates simulating N and S deposition of urban areas. Peroxidase activity in the fine roots of seedlings planted in polluted forest soils was higher than in those planted in non-polluted soils and correlated positively with the activities measured in an earlier study in the roots of mature Scots pines growing at the sites from where the soils were collected. Growth of seedlings and mycorrhizal status were not affected by the origin of soil. Exposing the seedlings to winter acclimation conditions for 6 weeks elevated peroxidase activity in the roots. The addition of ammonium or potassium sulphate to non-polluted soils did not induce elevated root peroxidase activity, although at the levels of 0.5 and 1.0 g of ammonium sulphate a slight increasing trend was observed. We suggest, that indirect biotic factors, i.e. changes in the community structure of soil fungi, early stages of recognition, and defence reactions of pine roots against saprophytic and pathogenic fungi may be participating in the elicitation of peroxidase (POD) activity, although the possible role of heavy metals cannot be excluded.     

Limited effects of heavy metal pollution on foraging and breeding success in the curlew (Numenius arquata)

We examined the effect of heavy metal pollution on the foraging success and breeding performance of the curlew (Numenius arquata) along a documented pollution gradient from a point source, and also by comparing foraging and breeding parameters between the polluted site and two non-polluted areas. Prey biomass and abundance, and foraging success did not vary along the pollution gradient, and were no less at the polluted site than in the non-polluted areas. Furthermore, there was no difference in adult weight during incubation between the polluted and non-polluted areas. There were also no differences in measures of breeding success along the pollution gradient, or between areas, which could be attributed to pollution per se. Egg shells from the polluted area had higher concentrations of heavy metals than in the non-polluted areas, and egg shells close to the pollution source were contaminated more than those further from it. However, there was no difference in calcium concentrations of egg shells or egg-shell thickness between areas. We conclude that in this study there were minimal immediate effects of heavy metal pollution on foraging and breeding success in the curlew.     

Breeding performance of blue tits (Cyanistes caeruleus) and great tits (Parus major) in a heavy metal polluted area

We compared heavy metal levels, calcium levels, breeding parameters and condition of nestling and adult Cyanistes caeruleus and Parus major along a heavy metal pollution gradient. Both species started laying earlier and showed inferior nestling growth and smaller fledging probability in the polluted areas, which are phenologically advanced in spring due to sparse forests. The major inter-specific difference in the responses was that the clutch size and hatching success were decreased in the polluted area in P. major, but not in C. caeruleus. Heavy metal profiles in nestling feces were relatively similar in the two species, though Ni and Pb levels were higher in C. caeruleus than in P. major. However, the latter species showed markedly higher fecal calcium concentrations. Lower calcium levels and higher levels of some heavy metals in C. caeruleus suggest that in Ca-deficient environments this species might be more susceptible to negative pollution effects than P. major.