Long-term Hg pollution-induced structural shifts of bacterial community in the terrestrial isopod (Porcellio scaber) gut

In previous studies we detected lower species richness and lower Hg sensitivity of the bacteria present in egested guts of Porcellio scaber (Crustacea, Isopoda) from chronically Hg polluted than from unpolluted environment. Basis for such results were further investigated by sequencing of 16S rRNA genes of mercury-resistant (Hg^r) isolates and clone libraries. We observed up to 385 times higher numbers of Hg^r bacteria in guts of animals from polluted than from unpolluted environment. The majority of Hg^r strains contained merA genes. Sequencing of 16S rRNA clones from egested guts of animals from Hg-polluted environments showed elevated number of bacteria from Pseudomonas, Listeria and Bacteroidetes relatives groups. In animals from pristine environment number of bacteria from Achromobacter relatives, Alcaligenes, Paracoccus, Ochrobactrum relatives, Rhizobium/Agrobacterium, Bacillus and Microbacterium groups were elevated. Such bacterial community shifts in guts of animals from Hg-polluted environment could significantly contribute to P. scaber Hg tolerance.     

Avoidance bio-assays may help to test the ecological significance of soil pollution

We measured the short-term (100 min) avoidance of a soil heavily polluted by hydrocarbons by the soil springtail Folsomia candida, at six rates of dilution in a control, unpolluted soil. We compared the results with those of long-term (40-day) population tests. Five strains were compared, of varying geographical and ecological origin. When pure, the polluted soil was lethal in the long-term and avoided in the short-term by all strains. Avoidance tests, but not population tests, were able to discriminate between strains. Avoidance thresholds differed among strains. Two ecological consequences of the results were discussed: (i) toxic compounds may kill soil animals or deprive them from food, resulting in death of populations, (ii) pollution spots can be locally deprived of fauna because of escape movements of soil animals. Advantages and limitations of the method have been listed, together with proposals for their wider use in soil ecology and ecotoxicology.     

A review on the application of microbial toxicity tests for deriving sediment quality guidelines

The results of microbial toxicity tests are needed for the risk assessment of polluted sediments. In comparison with animals the anaerobic microorganisms are more tolerant to natural sediment conditions whereas they are more sensitive for a number of specific pollutants. Microbial toxicity tests from a literature search were classified in seven categories. Category A, B and C use polluted sediments and are applied for sediment monitoring. In category D, a pure chemical is added and the organisms and the test conditions were derived from sediment. Therefore this category can be used for setting sediment quality guidelines which protect sediment functions for the toxic effects of chemicals. In category E, organisms from a polluted site are separated from the sediment and are tested with pure chemicals. Organisms from a more polluted site can be more tolerant to a local pollutant. This is called pollution-induced community tolerance and can be used as evidence for the occurrence of toxic effects in a specific sediment. In category F pure chemicals are tested with a pure culture of microorganisms under sediment conditions. The results of category F tests can be combined with single species tests with animals and plants to obtain sediment quality guidelines sufficient for species protection. This can be compared with the sediment quality guidelines which protect sediment functions. When one of these quality guidelines is exceeded for a compound at a specific location a category E test can be used to determine whether the compound shows toxic effects in that sediment.     

Metal-induced tolerance in the freshwater microbenthic diatom Gomphonema parvulum

The benthic diatom Gomphonema parvulum Kützing is a common species in both clean and metal contaminated rivers. Our aim was to investigate whether metal-induced tolerance could explain the persistance of this taxon under metal polluted conditions. G. parvulum strains were isolated from a Zn- and Cd-contaminated stream and from a relatively clean ("reference") stream. The strains were cultured in synthetic medium as mono-specific biofilms to maintain their specific benthic growth features. Moreover, the strain from the metal polluted stream was cultured in plain and Zn- and Cd-enriched synthetic medium. Short-term (5 h) toxicity experiments with Zn were performed with the strains using pulse amplitude modulated (PAM) fluorometry. Zn lowered significantly the minimal chlorophyll fluorescence (F0) and the photon yield (phi(p)) of the exposed strains after 5 h exposure. The actual Zn concentrations that caused a 50% reduction (EC50's) of the phi(p) of the strain from the metal polluted stream were significantly higher than those of the isolate from the unpolluted stream. The absence of tolerance to Cu of the "polluted" strain indicated that Zn tolerance resulted from specific induction by chronic exposure to Zn in the field. Observations on field biofilms confirmed a higher tolerance of the G. parvulum population from the polluted stream than of the G. parvulum population from the reference stream. A genetic nature of this metal adaptation was supported by the persistance of the Zn tolerance of the polluted strain 2 years after isolation.     

Experimental exposure to cadmium affects metallothionein-like protein levels but not survival and growth in wolf spiders from polluted and reference populations

Both local adaptation and acclimation in tolerance mechanisms may allow populations to persist under metal pollution. However, both mechanisms are presumed to incur (energetic) costs and to trade-off with other life-history traits. To test this hypothesis, we exposed Pardosa saltans (Lycosidae) spiderlings originating from metal-polluted and unpolluted sites to a controlled cadmium (Cd) treatment, and compared contents of metal-binding metallothionein-like proteins (MTLPs), internal metal concentrations, and individual survival and growth rates with a reference treatment. While increased MTLP concentrations in offspring originating from both polluted and unpolluted populations upon exposure indicates a plastic tolerance mechanism, survival and growth rates remain largely unaffected, independent of the population of origin. However, MTLP and Cd concentrations were not significantly correlated. We suggest that MTLP production may be an important mechanism enabling P. saltans populations to persist in ecosystems polluted with heavy metals above a certain level.     

Effects of pollution on humic substances

To assess effects of industrial and environmental pollution on analytical characteristics of humic substances, we isolated humic acids (HA's) and fulvic acids (FA's) from unpolluted and polluted soils and sediments. Following purification, the HA's and FA's were characterized by elemental (C, H, O, N, S) and functional group (CO2H, phenolic OH, total acidity) analyses, infrared (IR) spectrophotometry, differential thermal analysis (DTA) and by metal (Fe, Al, Cu, Mn, Pb, Ni, Co, Zn, Cr, Cd, Hg, Ca and Mg) analyses. Si was also determined in all samples. Polluted HA's and FA's contained more N and S but less O and were richer in all metals and Si than were unpolluted ones. IR spectra showed that polluted humic materials were enriched in COO- groups, secondary non-cyclic amides and possible also in SO3H groups. DTA curves indicated that polluted HA's and FA's were more thermostable than unpolluted HA's and FA's. Unusually high N, S, Cu, Cr, Zn and Hg contents of humic materials appear to be useful indicators of soil and sediment pollution.     

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.     

Effects of pollution on humic substances

Abstract

To assess effects of industrial and environmental pollution on analytical characteristics of humic substances, we isolated humic acids (HA's) and fulvic acids (FA's) from unpolluted and polluted soils and sediments. Following purification, the HA's and FA's were characterized by elemental (C, H, O, N, S) and functional group (CO₂H, phenolic OH, total acidity) analyses, infrared (IR) spectrophotometry, differential thermal analysis (DTA) and by metal (Fe, Al, Cu, Mn, Pb, Ni, Co, Zn, Cr, Cd, Hg, Ca and Mg) analyses. Si was also determined in all samples.

Polluted HA's and FA's contained more N and S but less 0 and were richer in all metals and Si than were unpolluted ones. IR spectra showed that polluted humic materials were enriched in COO^‐ groups, secondary non‐cyclic amides and possible also in SO₃H groups. DTA curves indicated that polluted HA's and FA's were more thermostable than unpolluted HA's and FA's. Unusually high N, S, Cu, Cr, Zn and Hg contents of humic materials appear to be useful indicators of soil and sediment pollution.     

ELISA detection of multixenobiotic resistance transporter induction in indigenous freshwater Chironomidae larvae (Diptera): A biomarker calibration step for in situ monitoring of xenobiotic exposure

A new simple and sensitive method to distinguish chemically polluted from unpolluted situations in freshwater ecosystems is reported. For this purpose, Chironomus gr thumni larvae were collected from a polluted urban river downstream a sewage treatment plant. For the first time, ELISA assay was used to semi-quantify the multixenobiotic resistance transporters (MXR) in these small pertinent bioindicators. The use of samples immediately fixed in the field gives a delay to isolate larvae and allows multi-sampling along a longitudinal transect in a river at a given time. Results exhibit an induction of MXR proteins in larvae from the polluted river and a deinduction in larvae maintained 11days in unpolluted water. They show new evidences to use midge larvae in biomonitoring environmental programs. They answer to first biomarker calibration steps for the ongoing development of MXR transporters as a detection tool of xenobiotic impacts on bioindicator invertebrates in their freshwater habitats.     

Purification of water contaminated with Hg using horizontal subsurface constructed wetlands

As a global pollutant, Hg (Hg) since the turn of the last century has received increased attention. Decreasing the emission of Hg into the food chain and the atmosphere is an effective way to reduce the Hg damage. The current study provided information about pilot-scale horizontal subsurface flow (HSSF) constructed wetlands (CWs) to remove different Hg species in polluted water. Synthetic wastewater was fed to two HSSF CWs, one was planted with Acorus calamus L and the other was unplanted as a control. The total Hg (THg), dissolved Hg (DHg), and particulate Hg (PHg) from five sites along the HSSF CWs were analyzed to describe the process of Hg removal. Results show that the CWs have high removal efficiency of Hg which is more than 90%. The removal efficiencies of THg and DHg from the unplanted CW were 92.1?±?3.6% and 72.4?±?13.1%, respectively. While, the removal efficiencies of THg and DHg in planted CW were 95.9?±?7.5% and 94.9?±?4.9%, which were higher than that in blank CW. The PHg was mainly removed in the first quarter of the CWs, which was also revealed by the partition coefficient K_d. To a certain extent, the effect of plants depends on the hydraulic retention time (HRT). The results in the current study show the potential of the HSSF-CWs for restoration from Hg-contaminated water.

     

Lead impact on nutrition, energy reserves, respiration and stress protein (hsp 70) level in Porcellio scaber (Isopoda) populations differently preconditioned in their habitats

The impact of lead on food consumption, energy metabolism and the stress protein (hsp 70) level was investigated in the woodlouse Porcellio scaber (Isopoda), a common representative of the saprophagous soil macrofauna. To examine possible acclimation or tolerance to lead in woodlice from a contaminated habitat, animals of two populations, one deriving from a lead-contaminated artillery range and one from an uncontaminated control stand, were exposed to a series of lead concentrations under otherwise constant laboratory conditions for a maximum of 80 days. The applied lead concentrations (at a maximum 7945 mg/kg food dry wt) did not have any significant quantitative effect on the food consumption of the isopods, although the population pre-exposed in the artillery range showed a tendency toward a higher food uptake than the control population. After 80 days of exposure, both populations showed an equal trend toward increasing their respiration as lead concentrations, that they had been fed on, were increased. Accordingly, the glycogen content of the body, in both populations, was elevated with increasing lead concentrations in the food. This effect was more pronounced in the pre-exposed isopod population than in the one from the control stand. The non-pre-exposed isopods showed a general tendency toward a lower protein content of their bodies than the pre-exposed ones, although no effect of the lead on this parameter could be statistically proven. The ability of the artillery range isopods to synthesise stress proteins (hsp 70) in response to lead contamination decreased at much lower lead concentrations in their food than in the non-pre-exposed control population, even though the artillery range isopods seemed to be equally or even slightly better equipped with energy storage products. Even though the better nutrient status of these animals might refer to some lead tolerance of the pre-exposed population, the stress protein data suggest that a metal-resistant Porcellio population did not evolve in this lead-contaminated site.     

Studies on the progeny of rice plants grown at an unpolluted and polluted site

Oryza sativa L. cv. GR3 was grown near to a fertilizer plant and the growth of its seed progeny was evaluated at sites near to, and distant from, the fertilizer plant. The grain yield was reduced in plants grown at the polluted site, with reduced panicle length and increased sterility index. This decrease in grain yield was higher in the progeny plants than in the normal polluted plants. The polluted progeny grown at a control site showed a normal yield, but an increased sterility index. Accumulation of total sulphur and fluoride was higher in the leaves of polluted rice plants. It was evident that tolerance to air pollution was reduced in the progeny plants due to an additional dosage of sulphur and fluoride pollutants and to unfavourable climatic factors. Further, pollution effects were not carried over in progeny plants when grown in an unpolluted environment.     

The relationship between metal composition,phenolic acid and flavonoid content in Imleria badia from non-polluted and polluted areas

ABSTRACT

The aim of this study was to determine the elemental composition, phenolic content and composition and antioxidant properties of Imleria badia (Fr.) Vizzini (former names Boletus badius (Fr.) Fr., and Xerocomus badius (Fr.) E.-J. Gilbert) fruiting bodies collected from sites with different levels of pollution. Imleria badia was relatively tolerant to soil contamination with toxic elements and was able to grow in As, Cd, Hg and Pb concentrations exceeding 15, 2.9, 0.4 and 77 mg kg^?1, respectively. The concentration of elements in soil was reflected in the element content in I. badia. The fruiting bodies from polluted sites exhibited significantly higher content of all the analyzed elements. Among 21 individual phenolic compounds only protocatechiuc and caffeic acids, and quercetin were determined in fruiting bodies of I. badia. The differences between the concentration of the quantified phenolic compounds and the total flavonoid content in fruiting bodies of I. badia from unpolluted and polluted sites were not significant. However, the greatest total phenolic content was found in fruiting bodies from the polluted areas. The antioxidative capacity of mushrooms collected from heavily polluted sites was lower than those growing in unpolluted areas. The concentrations of some metals in soil and fruiting soil were positively correlated with phenolic content and IC₅₀.     

Seasonal changes in the sulphate content of deciduous woodland soils exposed to atmospheric pollution

Seasonal changes in soil pH, sulphate concentration and total-S were measured in two brown earth soils, sampled from deciduous woodlands. One site studied was exposed to severe atmospheric pollution from a coking works while the other site was relatively unpolluted but located in an area receiving wet and dry deposited acidity of greater than 1.0 and 2.4 kg H(+) ha(-1) year(-1), respectively The pH of soil at the heavily polluted site was lower than the relatively unpolluted soil at each monthly sample point, except during November. Annual average sulphate concentrations (LiCl-extractable) were highest in the soil exposed to coking pollution, where they peaked during summer and autumn. A marked difference in total-S was found in soils from the two sites, the heavily polluted soil showing the highest concentration with peaks again occurring during late summer and autumn. Only 4.0% (w/w) of the total-S of the heavily polluted soil occurred as LiCl-extractable sulphate, compared to 21.4% (w/w) for the relatively unpolluted soil, showing that organic sulphur is increased in brown earths following exposure to severe atmospheric pollution from the coking works.     

The sea–air exchange of mercury (Hg) in the marine boundary layer of the Augusta basin (southern Italy): Concentrations and evasion flux

The first attempt to systematically investigate the atmospheric mercury (Hg) in the MBL of the Augusta basin (SE Sicily, Italy) has been undertaken. In the past the basin was the receptor for Hg from an intense industrial activity which contaminated the bottom sediments of the Bay, making this area a potential source of pollution for the surrounding Mediterranean. Three oceanographic cruises have been thus performed in the basin during the winter and summer 2011/2012, where we estimated averaged Hg_atm concentrations of about 1.5 ± 0.4 (range 0.9–3.1) and 2.1 ± 0.98 (range 1.1–3.1) ng m⁻³ for the two seasons, respectively. These data are somewhat higher than the background Hg_atm value measured over the land (range 1.1 ± 0.3 ng m⁻³) at downtown Augusta, while are similar to those detected in other polluted regions elsewhere. Hg evasion fluxes estimated at the sea/air interface over the Bay range from 3.6 ± 0.3 (unpolluted site) to 72 ± 0.1 (polluted site of the basin) ng m⁻² h⁻¹. By extending these measurements to the entire area of the Augusta basin (∼23.5 km²), we calculated a total sea–air Hg evasion flux of about 9.7 ± 0.1 g d⁻¹ (∼0.004 t yr⁻¹), accounting for ∼0.0002% of the global Hg oceanic evasion (2000 t yr⁻¹). The new proposed data set offers a unique and original study on the potential outflow of Hg from the sea–air interface at the basin, and it represents an important step for a better comprehension of the processes occurring in the marine biogeochemical cycle of this element.     

Heavy metal accumulation, heat shock protein expression and cytogenetic changes in Tetrix tenuicornis (L.) (Tetrigidae, Orthoptera) from polluted areas

The orthopteran insect Tetrix tenuicornis, collected from polluted and unpolluted areas, was used to study heavy metal accumulation and its impact on stress protein levels and on changes in the number and morphology of chromosomes in mitotic and meiotic cells. During two consecutive years, insects were collected from polluted areas of zinc-lead mine spoils near Boles?aw (Poland) and from unpolluted areas near Busko and Staszów (Poland). T. tenuicornis from the polluted area showed 1.5, 4.03, 4.32 and 41.73 times higher concentrations of copper (Cu), zinc (Zn), lead (Pb) and cadmium (Cd), respectively, than insects of the same species collected from unpolluted areas. Insects exposed to heavy metals showed only small changes, and rather a decrease in the concentration of constitutive and inducible heat shock proteins Hsp70, the level of which increases under stress conditions. A cytogenetic study of T. tenuicornis revealed intra-population anomalies in chromosome number and morphology in mitotic and meiotic cells and the presence of an additional B chromosome in germinal cells. In 50% of females collected from polluted areas, mosaic oogonial mitotic chromosome sets and diploid, hypo- or hypertetraploid, tetraploid, and octoploid chromosome numbers were detected. In turn, 14.6% of males showed a heterozygous deficiency of chromatin in L2 and M3 bivalents in addition to the presence of B chromosomes.     

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.     

Concentration of bioactive compounds in mussels Mytilus galloprovincialis as an indicator of pollution

The polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and organotins were analyzed in mussels Mytilus galloprovincialis from polluted and unpolluted sites from Mokpo Bay, Korea. The total PAH's concentrations (10(-3)mgkg(-1)) measured by GC-MS were in the range from 31+/-23 to 1+/-1. Among the eight PAHs the predominant ones were fluoranthene, phenanthrene and pyrene and accounted approximately 63% of the total PAHs. Among the four detected PCBs the highest content was of PCB 153, which accounted about 47% of the total PCBs. The main organotin compounds were dibutyltindichloride (DBT) and tributyltinchloride (TBT) and their composition was approximately 33% and 24%. PAHs, PCBs and organotins were found only in the mussels from polluted site. The antioxidant activity by ABTS [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)] test was higher in mussels from polluted than from unpolluted sites (P<0.05). It was found a correlation between the determined compounds (PAHs, PCBs and organotins) and the antioxidant activity of the mussel tissue from polluted site and the correlation coefficients were 0.96, 0.92 and 0.80, respectively. Such correlation can be explained by the properties of mussels. Since the mussel cell wall and tissues are hydrophobic, they can concentrate a number of hydrophobic pollutants like PAHs and PCBs from the marine environment by solubility rules. On the other hand, proteins are lipophilic compounds having antioxidant properties. Certain amino acid residues and thiol (-SH) groups, contained in proteins, respond to the ABTS antioxidant activity assay. Thus there may be a correlation between the total antioxidant activity of the organism and the PAH-PCB pollutants which were concentrated from its environment. The studied properties of mussels from polluted site can be used as an additional indicator of pollution.     

Heavy metal accumulation in Littoraria scabra along polluted and pristine mangrove areas of Tanzania

The periwinkle Littoraria scabra was collected at polluted and pristine mangrove sites along the Tanzanian coastline, including Msimbazi, Mbweni (i.e. Dar es Salaam) and Kisakasaka, Nyamanzi and Maruhubi (i.e. Zanzibar). Periwinkles were morphologically characterized, sexed and their heavy metal content was determined using ICP-MS. Analysis revealed that L. scabra from polluted areas contained higher soft tissue heavy metal levels, were significantly smaller and weighed less compared to their conspecifics from the unpolluted mangroves. The current morphological observations may be explained in terms of growth and/or mortality rate differences between the polluted and non-polluted sites. Although a variety of stressors may account for these adverse morphological patterns, our data suggest a close relationship with the soft tissue heavy metal content. Compared to soft tissue heavy metal levels that were measured in L. scabra along the same area in 1998, most metals, except for arsenic, chromium and iron have decreased dramatically.     

Absence of Hg transpiration by shoot after Hg uptake by roots of six terrestrial plant species

In this paper we investigated if, and to what extent, six different plant species accumulate, translocate and emit mercury (Hg) into the air. The Hg uptake by roots, distribution of Hg to the shoot and release of Hg via shoots of garden pea, spring wheat, sugar beet, oil-seed rape, white clover and willow were investigated in a transpiration chamber. The airborne Hg was trapped in a Hopcalite trap or a gold trap. Traps and plant materials were analysed for content of Hg by CVAAS. The results show that all plant species were able to take up Hg to a large extent from a nutrient solution containing 200 microg L(-1) Hg. However, the Hg translocation to the shoot was low (0.17-2.5%) and the Hg that reached the leaves was trapped and no release of the absorbed Hg to the air was detected.