Heavy metal tolerance by ectomycorrhizal fungi and metal amelioration by Pisolithus tinctorius

Five ectomycorrhizal fungi, Pisolithus tinctorius, Thelephora terrestris, Cenococcum geophilum, Hymenogaster sp. and Scleroderma sp., which were demonstrated previously to be capable of forming ectomycorrhizas with some pine, eucalypt and fagaceous tree species were grown in vitro in liquid cultures for 3 weeks at six different concentrations of nine heavy metals, aluminium, iron, copper, zinc, nickel, cadmium, chromium, lead and mercury. Measurements of mean mycelial dry weight yields indicated that the local isolates of Hymenogaster sp. and Scleroderma sp., as well as the introduced fungal species P. tinctorius, were able to withstand high concentrations of Al, Fe, Cu and Zn and might, therefore, have potential for revegetation schemes in metal-contaminated soils. The metal amelioration mechanism in the metal-tolerant fungal species P. tinctorius was observed to involve extrahyphal slime and, as demonstrated by energy-dispersive X-ray spectrometry, was achieved by polyphosphate linkage of Cu and Zn.     

Salicornia spp. as a biomonitor of Cu and Zn in salt marsh sediments

Macroalgae in estuarine and coastal waters, in contrast to vascular salt marsh plants, have previously been utilised as biomonitors of sediment-held metals. The colonising halophyte Salicornia spp., however, occurs in both mudflats alongside macroalgae, as well as in association with salt marsh vascular plants. The present research aims to determine the relationships between fluctuations in sediment-held metals and those in Salicornia spp. over the course of a growing season. Samples of the species and corresponding underlying sediment were collected from the metal mine-polluted Restronguet Creek of the Fal Estuary, Cornwall on a monthly basis between March and November, 2000. Oven-dried sediment and vegetation samples were analysed for total Fe, Cu, Zn and Mn. Significant correlations with both the roots and aerial portion of the plant were found with sediment Cu and Zn concentrations. Significant relationships with either Mn or Fe were not observed. Thus, Salicornia spp. would appear to be a suitable tool for biomonitoring Zn and, particularly, Cu. Hyperaccumulation of Zn in the aerial portion during initial growth also indicates that Salicornia spp. may be useful for alleviating metal contamination through phytoextraction, whilst Cu in the roots is proposed as having potential for phytostabilization.     

Trace element levels in mollusks from clean and polluted coastal marine sites in the Mediterranean, Red and North Seas

The trace element contamination levels in mollusks were evaluated for different marine coastal sites in the Mediterranean (Israeli coast), Red (Israeli coast) and North (German coast) Seas. Three bivalve species (Mactra corallina, Donax sp, and Mytilus edulis) and two gastropod species (Patella sp.and Cellana rota) were sampled at polluted and relatively clean sites, and their soft tissue analyzed for Hg, Cd, Zn, Cu, Mn and Fe concentrations. Representative samples were screened for organic contaminants [(DDE), polychlorinated biphenyls PCBs and polycyclic aromatic hydrocarbons (PAHs)] which exhibited very low concentrations at all sites. In the Red Sea, the gastropod C. rota showed low levels of Hg (below detection limit) and similar Cd concentrations at all the examined sites, while other trace elements (Cu, Zn, Mn, Fe) were slightly enriched at the northern beach stations. Along the Mediterranean coast of Israel, Hg and Zn were enriched in two bivalves (M. corallina and Donax sp.) from Haifa Bay, both species undergoing a long-term decrease in Hg based on previous studies. Significant Cd and Zn enrichment was detected in Patella sp. from the Kishon River estuary at the southern part of Haifa Bay. In general, Patella sp. and Donax sp. specimens from Haifa Bay exhibited higher levels of Cd compared to other sites along the Israeli Mediterranean coast, attributed to the enrichment of Cd in suspended particulate matter. Along the German coast (North Sea) M. edulis exhibited higher concentrations of Hg and Cd at the Elbe and Eider estuaries, but with levels below those found in polluted sites elsewhere. Received: 25 February 1999 / Received in revised form: 22 April 1999 / Accepted: 30 April 1999     

Comparative strategies of heavy metal accumulation by crustaceans: zinc,copper and cadmium in a decapod,an amphipod and a barnacle

This study investigates the comparative strategies of accumulation under standardised laboratory conditions of the essential metals zinc and copper, and the non-essential metal cadmium by three crustaceans of different taxa; vizPalaemon elegans Rathke (Malacostraca: Eucarida: Decapoda),Echinogammarus pirloti (Sexton & Spooner) (Malacostraca: Peracarida: Amphipoda) and the barnacleElminius modestus Darwin (Cirripedia: Thoracica).The decapodP. elegans regulates body zinc concentrations to a constant level (ca. 79 µg Zn g^–1) over a wide range of dissolved metal availabilities until regulation breaks down at high Zn availabilities and net accumulation begins. The amphipodE. pirloti accumulates zinc at all dissolved zinc concentrations but at a low net rate such that the accumulation strategy approaches that of regulation. The barnacleE. modestus accumulates zinc to high body concentrations with no significant excretion of accumulated zinc. In the case of copper,P. elegans similarly regulates body copper concentrations to a constant level (ca. 129 µg Cu g^–1) over a range of dissolved copper availabilities until regulation breaks down at high copper concentrations. Both the amphipodE. pirloti and the barnacleE. modestus on the other hand accumulate copper at all dissolved copper exposures with no evidence of regulation. All three crustaceans accumulate the non-essential metal cadmium at all dissolved cadmium concentrations without regulation.Heavy metal accumulation strategies therefore vary between crustacean taxa and between metals. Uptake rates for zinc and cadmium have been estimated for the three crustaceans and can be interpreted in terms of cuticle permeability and way of life of each crustacean. Examination of these uptake rates provides an insight into possible reasons behind the adoption of particular metal accumulation strategies.     

Screening of metal uptake by plant colonizers growing on abandoned copper mine in Kapunda,South Australia

Systematic site survey for sample collection and analysis was conducted at a derelict copper (Cu) mine at Kapunda, South Australia. Cu concentrations in the soils at this former mine ranged from 65–10107 mg kg^?1. The pH and EC varied widely in the 3.9–8.4 and 152–7311 µS ranges, respectively. Nine plant species growing over the copper mine site were selected to screen for metal uptake to determine their suitability for phytoremediation. The Australian native tree species Eucalyptus camaldulensis indicated enrichment factor (EF) of 2.17, 1.89, and 1.30 for Cu, Zn, and Pb, respectively, suggesting that this species of tree can accumulate these metals to some degree. The stress-resistant exotic olive, Olea europaea exhibited EF of ≤ 0.01 for Cu, Cd, and Pb, and 0.29 for Zn, which is characteristic of an excluder plant. Acacia pycnantha, the Australian pioneer legume species with EF 0.03, 0.80, 0.32, and 0.01 for Cu, Zn, Cd, and Pb, respectively, emerged as another strong metal excluder and consequently as an ideal metal stabilizer.     

Relationship between Mikania micrantha invasion and soil microbial biomass, respiration and functional diversity

The aim of this study was to determine elemental composition of sap-feeding insects inhabiting various parts of the Ni hyperaccumulating plant Berkheya coddii Roessl., the endemic species of ultramafic outcrops in Mpumalanga, South Africa. Three species were examined: the aphid Protaphis pseudocardui (Aphididae), abundant on young leaves; the mealybug Orthesia sp. (Ortheziidae) colonizing underground parts of this plant, and the bug Norialsus berkheyae (Cixiidae) living on young shoots. Maps of Ni, K, Ca, Zn, and Fe for selected body areas of these species were generated using Dynamic Analysis method on the basis of particle-induced X-ray emission (micro-PIXE) and proton backscattering (BS) measurements. Atomic absorption spectrometry was used to determine Ni, Zn, Cu, Fe contents in the B. coddii organs, in some sap-feeding insect species including these mentioned above, and in the assassin bug hunting on Chrysolina pardalina, a monophagous beetle of B. coddii. Bioaccumulation factor for Ni in the examined species was below 0.05, and much higher for other metals (Zn ≥ 2; Fe ≤ 5). Ni distribution within body was species-dependent. It was the highest in the antennae of P. pseudocardui, in the head of Orthesia sp. and in the metathorax of N. berkheyae. Distribution patterns of other metals were different among examined species. Ca was recorded mainly in peripheral parts of the body in all species. Zn showed similar distribution to Ni. Fe distribution was similar to Ni only in the mealybugs. Uneven concentrations of metals within selected body regions indicated their relations with specific organs. Analysis of Ni transfer to higher trophic levels was done on the basis of two food nets: B. coddii—C. pardalina—Rhinocoris neavii and B. coddii—P. pseudocardui—Polyrhachis ant and led to the conclusion that the role of sap-feeding insects in Ni transfer was marginal.     

Seasonal variation of arbuscular mycorrhizal fungi in temperate grasslands along a wide hydrologic gradient

We studied seasonal variation in population attributes of arbuscular mycorrhizal (AM) fungi over 2 years in four sites of temperate grasslands of the Argentinean Flooding Pampas. The sites represent a wide range of soil conditions, hydrologic gradients, and floristic composition. Lotus glaber, a perennial herbaceous legume naturalised in the Flooding Pampas, was dominant at the four plant community sites. Its roots were highly colonised by AM fungi. Temporal variations in spore density, spore type, AM root colonisation, floristic composition and soil chemical characteristics occurred in each site and were different among sites. The duration of flooding had no effect on spore density but depressed AM root colonisation. Eleven different types of spores were recognized and four were identified. Two species dominated at the four sites: Glomus fasciculatum and Glomus intraradices. Spore density was highest in summer (dry season) and lowest in winter (wet season) with intermediate values in autumn and spring. Colonisation of L. glaber roots was highest in summer or spring and lowest in winter or autumn. The relative density of G. fasciculatum and G. intraradices versus Glomus sp. and Acaulospora sp. had distinctive seasonal peaks. These seasonal peaks occurred at all four sites, suggesting differences among AM fungus species with respect to the seasonality of sporulation. Spore density and AM root colonisation when measured at any one time were poorly related to each other. However, spore density was significantly correlated with root colonisation 3 months before, suggesting that high colonisation in one season precedes high sporulation in the next season.     

Variation in soft tissue chemistry among scyphozoan and cubozoan jellyfishes from the Great Barrier Reef,Australia

Bioaccumulation of trace elements in jellyfish has so far received little attention, despite their being prey for many animals from multiple trophic levels and targeted by commercial jellyfish fisheries. Scyphozoan and cubozoan jellyfish were collected over a three year period from across-shelf and along the northern and central Great Barrier Reef, Australia. To test the hypotheses that jellyfishes were able to accumulate elements above ambient background levels, and if there were spatial or temporal variations among species, soft tissue concentrations of 14 trace elements were compared with ambient seawater concentrations. Most elements, including aluminium, arsenic, barium, cadmium, chromium, copper, iron, manganese and zinc were measured at concentrations above ambient seawater levels indicating bioaccumulative capacity. Results showed some regulation of lithium in Cassiopea sp., Cyanea sp. and Mastigias sp., while calcium, magnesium and strontium reflected ambient conditions for all species. Accumulation varied significantly among species and sampling locations. For Mastigias sp. and Netrostoma sp., tissue concentrations of Al, As, Cu, Fe and Zn decreased with distance from the mainland. The hypothesis that jellyfishes are capable of accumulating trace elements was accepted, and their use as biomonitors should be investigated further.     

Relation of desert pupfish abundance to selected environmental variables in natural and manmade habitats in the Salton Sea basin

Synopsis We assessed the relation between abundance of desert pupfish, Cyprinodon macularius, and selected biological and physicochemical variables in natural and manmade habitats within the Salton Sea Basin. Field sampling in a natural tributary, Salt Creek, and three agricultural drains captured eight species including pupfish (1.1% of the total catch), the only native species encountered. According to Bray–Curtis resemblance functions, fish species assemblages differed mostly between Salt Creek and the drains (i.e., the three drains had relatively similar species assemblages). Pupfish numbers and environmental variables varied among sites and sample periods. Canonical correlation showed that pupfish abundance was positively correlated with abundance of western mosquitofish, Gambusia affinis, and negatively correlated with abundance of porthole livebearers, Poeciliopsis gracilis, tilapias (Sarotherodon mossambica and Tilapia zillii), longjaw mudsuckers, Gillichthys mirabilis, and mollies (Poecilia latipinnaandPoecilia mexicana). In addition, pupfish abundance was positively correlated with cover, pH, and salinity, and negatively correlated with sediment factor (a measure of sediment grain size) and dissolved oxygen. Pupfish abundance was generally highest in habitats where water quality extremes (especially high pH and salinity, and low dissolved oxygen) seemingly limited the occurrence of nonnative fishes. This study also documented evidence of predation by mudsuckers on pupfish. These findings support the contention of many resource managers that pupfish populations are adversely influenced by ecological interactions with nonnative fishes.     

Influence of heavy metal leaf contaminants on the in vitro growth of urban-tree phylloplane-fungi

The surfaces of urban woody vegetation are contaminated with varying amounts of numerous metallic compounds, including Cd, Cu, Mn, Al, Cr, Ni, Fe, Pb, Na, and Zn. To examine the possibility that these metals may affect phylloplane fungi, the above cations were tested in vitro for their ability to influence the growth of numerous saprophytic and parasitic fungi isolated from the leaves of London plane trees. Considerable variation in growth inhibition by the metals was observed. GenerallyAureobasidium pullulans, Epicoccum sp., andPhialophora verrucosa were relatively tolerant;Gnomonia platani, Cladsporium sp., andPleurophomella sp. were intermediate; andPestalotiopsis andChaetomium sp. were relatively sensitive to the incorporation of certain metals into solid and liquid media. If similar growth inhibitions occur in nature, competitive abilities or population structures of plant surface microbes may be altered by surface metal contamination. Metals causing the greatest and broadest spectrum growth suppression included Ni, Zn, Pb, Al, Fe, and Mn.     

Association of Copper and Zinc Levels in Oil Palm (Elaeis guineensis) to the Spatial Distribution of Ganoderma Species in the Plantations on Peat

Nutrients are essential for normal physiological processes in plants, and they play important roles in defence mechanisms against pathogens. Oil palms cultivated on peat are more prone to nutrient deficiency, especially micronutrients, and this may affect their susceptibility to Ganoderma species, the major threat to the sustainability of oil palm throughout South‐East Asia. This study was conducted to investigate the association of copper (Cu) and zinc (Zn) in mature oil palm to the spatial distribution of Ganoderma species in the plantations on peat. Foliar samples (frond 17) of oil palm from two plantations (Betong and Miri) on peat in Sarawak, Malaysia, were collected based on the spatial distribution pattern of Ganoderma, and total Cu and Zn were quantified spectrometrically. The experiment was conducted twice at a 1‐year interval. The concentrations of Cu and Zn were significantly lower in oil palms from infected areas in contrast to those from uninfected areas. In addition, oil palms in infected areas in Miri suffered Cu and Zn deficiencies. Furthermore, Cu and Zn were significantly lower in the oil palms in Miri that had higher Ganoderma occurrence, as compared to those in Betong, which had significantly higher Cu and Zn but lower Ganoderma occurrence.     

Phenolic substances of brown algae and their antioxidant activity

The composition and content of phenolic substances were studied in 14 species of marine brown algae of the Canary Islands littoral (Spain). The highest content of phenolic substances was found inCystoseira compressia andSargassum furcatum. A high antioxidant activity was found in florotannin isolated fromCystoseira sp.     

Benthos of Lake Mikri Prespa (North Greece)

An extensive sampling of Lake Mikri Prespa was carried out in summer 1990, aimed at evaluating benthic abundance, recording the species involved and detecting their response to some environmental variables. Three oligochaete species are endemic to the region,Potamothrix prespaensis, Psammoryctides ochridanus variabilis andPsammoryctides ochridanus typica. The bathymetric curve of the benthos resembles that of eutrophic lakes. A great abundance ofChironomus plumosus andChaoborus flavicans occurred. Four community patterns were revealed by TWINSPAN analysis, division level two, explicitly discriminated by the influence of substrate type, depth, C/N ratio and pH. Species richness and diversity was minimal in the profundal and maximal in the shallows, whilst the equitability index was high in all site groups. Depth and bottom type were the most important variables in defining species distribution. More precisely,Chironomus plumosus, Chaoborus flavicans andPotamothix hammoniensis were ubiquitous, preferring mostly deep waters.Tanytarsus sp.,Einfeldia dissidens, Cladopelma lateralis, Tanypus punctipennis andPotamothrix prespaensis favoured shallower and less productive waters with a bottom rich in decaying plant fragments, and thePsammoryctides species showed a littoral origin and regular occurrence on gravel beds. A few species displayed a particular distribution with changes of depth, C/N ratio and pH.     

Bioreduction of Cu(II) by Cell-Free Copper Reductase from a Copper Resistant <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. NA

Environmental copper contamination is a serious human health problem. Copper reductase is produced by microorganisms to facilitate copper uptake by ATPases into the cells increasing copper biosorption. This study assessed the reduction of Cu(II) by cell-free extracts of a highly copper-resistant bacterium, Pseudomonas sp. strain NA, isolated from vineyard soil contaminated with copper. Both intact cells and cell-free extract of Pseudomonas sp. strain NA displayed substantial reduction of Cu(II). Intact cells reduced more then 80 mg L⁻¹ of Cu(II) from medium amended with 200 mg L⁻¹ of copper after 24 h of incubation. Cell-free extract of the isolate reduced more than 65% of the Cu(II) at initial copper concentration of 200 mg L⁻¹ after 24 h. Soluble protein production was high at 72 h of incubation at 100 mg L⁻¹ of copper, with more then 60 μg L⁻¹ of total soluble protein in cell-free extract recorded. Cu(II) reduction by isolate NA was increased when copper concentration increased for both intact cells and cell-free extract. Results indicate that Pseudomonas sp. strain NA produces copper reductase enzyme as the key mechanism of copper biotransformation.     

Effects of Acid Mine Drainage on the Estuarine Food Web, Britannia Beach, Howe Sound, British Columbia, Canada

We investigated the effects of acid mine drainage (AMD) from Britannia Creek, BC, Canada, a stream contaminated by an abandoned copper mine, on the estuarine food web. Near the mouth of Britannia Creek, sediments and the water contained high concentrations of dissolved copper and the waters were highly acidic. Relative to a reference location, rockweed (Fucus gardneriSilva) cover was reduced, phytoplankton biomass was lower, chironomid larvae were less abundant, and fewer gammarid amphipods colonized basket traps. Laboratory toxicity tests confirmed that sediments and water from the lower reaches of Britannia Creek and its estuary were highly toxic to midge larvae (Chironomus tentansFabricius), the marine amphipod (Eohaustorius washingtonianusThorsteinson) and also impaired fertilization of sand dollars (Dendraster excentricusEschscholtz). Analyses of stomach contents of chum salmon fry (Oncorhynchus ketaWalbaum) indicated that they preferred to feed on chironomid larvae and gammarid amphipods at Britannia Creek estuary but these two taxa were less abundant at Britannia Creek estuary relative to the reference location. AMD from Britannia Mine disrupted the structure and function of the intertidal ecosystem resulting in loss of food production for fish including chum salmon fry and other species.     

Relationship between copper- and zinc-induced oxidative stress and proline accumulation in<Emphasis Type="Italic"> Scenedesmus</Emphasis> sp.

A 4-h exposure of Scenedesmus sp. to Cu or Zn enhanced intracellular levels of both test metals and proline. The level of intracellular proline increased markedly up to 10 µM Cu, but higher concentrations were inhibitory. However, intracellular proline consistently increased with increasing concentration of Zn in the medium. Cu and Zn induced oxidative stress in the test alga by increasing lipid peroxidation and membrane permeability, and by reducing SH content. Pretreatment of the test alga with 1 mM proline for 30 min completely alleviated Cu-induced lipid peroxidation, minimized K⁺ efflux and also reduced depletion of the SH pool. But proline pretreatment could only slightly reduce Zn-induced oxidative stress. Interestingly, proline pretreatment increased the level of Cu (25–54%) and Zn (19–49%) inside the cells. It did not affect the activities of superoxide dismutase, ascorbate peroxidase or catalase, but improved glutathione reductase activity under Cu and Zn stress. A comparison of the effects of proline pretreatment on lipid peroxidation by Cu, Zn, methyl viologen and ultraviolet-B radiation suggests that proline protects cells from metal-induced oxidative stress by scavenging reactive oxygen species rather than by chelating metal ions. Pretreatment of cells with a known antioxidant (ascorbate) and a hydroxyl radical scavenger (sodium benzoate) considerably reduced metal-induced lipid peroxidation and proline accumulation. However, sodium benzoate had a very mild effect on Zn-induced lipid peroxidation and proline accumulation. The present study demonstrates that proline possibly acts by detoxifying reactive oxygen species, mainly hydroxyl radicals, rather than by improving the antioxidant defense system under metal stress.Abbreviations APOX Ascorbate peroxidase - CAT Catalase - GR Glutathione reductase - MDA Malondialdehyde - MV Methyl viologen - ROS Reactive oxygen species - SH Sulphydryl - SOD Superoxide dismutase - UV-B Ultraviolet-B radiation     

Spatial Distribution of Heavy Metals in Soil and Flora Associated with the Glass Industry in North Central India: Implications for Phytoremediation

The effect of the glass industry on soil metal characterization was assessed at five test sites at five successive distances in a semi-arid area. A comprehensive profile of Zn, Cd, Pb, Ni, Cu, and As levels in soils was obtained. The spatial distribution patterns of integrated contamination indices for these metals show a similar decreasing trend in distribution as we move further from the industrial cluster. There was significant correlation among individual heavy metal concentrations in the soil samples. Integrated contamination indices indicate that 64% of the sites were in the high contamination range and 28% were in the moderate contamination range. A statistically significant difference (P ≤ 0.001) was obtained for each metal on comparing mean metal content among soil samples. Phytoremedial potential of 12 native plant species was also evaluated. Individual elements displayed remarkably different patterns of accumulation in soils as well as plants. Plants established limited capabilities in mobilizing Zn, Pb, Ni, and Cu in the root zone. While Cd, Cu, As, Zn and Pb were predominantly partitioned in shoots, Ni was equally partitioned between shoots and roots. Interestingly, some plants showed a different partitioning trend at higher concentrations of different metals compared to lower concentrations. Potential species for phytoremediation include Calotropis procera (Milk weed), Chenopodium murale (Goosefoot),Poa annua (Annual bluegrass) and Datura stramonium (Thorn apple). None of the species showed phytoremedial potential for Ni and Cu.     

Selective adaptation to copper of populations ofAgrostis tenuis andFestuca rubra (Poaceae)

Populations ofAgrostis Tenuis andFestuca rubra, tolerant and non-tolerant towards Zn and Pb, were studied in regard to their adaptability by natural selection, towards a third metal (Cu) and the time required for the appearance of such tolerance. It was found that Zn and Pb tolerant and non-tolerant populations are likely to select the character of Cu-tolerance within the course of only one generation. More specifically, Zn and Pb tolerant populations show a better adaptability compared with non-tolerant ones. The amount of tolerance to copper obtained by artificial selection is comparable with that of populations growing naturally on copper mines.