Effects of Heavy Metal Pollution on Oak Leaf Microorganisms

During the growing season, comparisons were made of the leaf surface microflora of (i) two groups of mature oak trees, one in the vicinity of a smelting complex contaminated by heavy metals and the other at a relatively uncontaminated site, and (ii) two groups of oak saplings at the uncontaminated site, one of which was sprayed with zinc, lead, and cadmium to simulate the heavy metal pollution from the smelter without the complicating effects of other pollutants. Total viable counts of bacteria, yeasts, and filamentous fungi (isolated by leaf washing) were generally little affected by the spraying treatment, whereas polluted leaves of mature trees supported fewer bacteria compared with leaves of mature trees at the uncontaminated site. Numbers of pigmented yeasts were lower on polluted oaks and on metal-dosed saplings compared with their respective controls. Polluted leaves of mature trees supported both greater numbers of Aureobasidium pullulans and Cladosporium spp. and a greater percentage of metal-tolerant fungi compared with oak leaves at the uncontaminated site. There were no significant overall differences in the degree of mycelial growth between the two groups of saplings or the mature trees.     

The assessment of soil contamination by heavy metals using geostatistical sequential Gaussian simulation method

Soil contamination by heavy metals is continuously increasing with a great attention in the world. Because, these elements negatively affect living life and the ecosystem. A total of 652 surface soil samples were collected at 100, 200, 400 m regular grid intervals from the study area in Turkey. The observed data does not show normal distribution. Cell declustering was done due to fact that data are not normal. Directional experimental semivariogram of the Cu and Ni showed that both geometric and zonal anisotropy exists in the data. Pb, Zn, Cr and Ba are qualified with omnidirectional experimental semivariogram models. The semivariograms characterized by spherical and Gaussian models of the elements were achieved. Geostatistical sequential Gaussian simulation (SGS) was applied to the study. A hundred simulated realizations depicted the spatial distribution and uncertainty of the elements in the site using a probabilistic approach which produced maps of the heavy metals. These maps showed contaminated and uncontaminated areas in the study site. The results revealed that 23%, 27%, and 24% of the study area at 60% probability were contaminated by the heavy metals including Cu, Cr, and Ni respectively. SGS results have been verified by a number of tests.     

Enhancing Degradation of Total Petroleum Hydrocarbons and Uptake of Heavy Metals in a Wetland Microcosm Planted with Phragmites Communis by Humic Acids Addition

The effects of humic acid (HA) on heavy-metal uptake by plants and degradation of total petroleum hydrocarbons (TPHs) in a wetland microcosm planted with Phragmites communis were evaluated by comparing waterlogged soils and water-drained upland soils. Experiments were conducted on soils artificially contaminated with heavy metals (Pb, Cu, Cd, Ni) and diesel fuel. HA showed a positive influence on biomass increase for all conditions, but more for belowground than aboveground biomass, and lower in contaminated than uncontaminated soil. The bioavailability and leachability factor (BLF) for all heavy metals except Ni increased with HA addition in both the control and the P. communis planted microcosms, suggesting that more heavy metals could be potentially phytoavailable for plant uptake. Microbial activities were not affected by both heavy metals and TPH contamination, and HA effects on stimulating microbial activities were much greater in the contaminated soil than under uncontaminated conditions. HA addition enhanced the degradation of TPH and n-alkane in waterlogged conditions. The results show that HA can increase the remedial performance in P. communis dominated wetlands simultaneously contaminated with heavy metals and petroleum hydrocarbons and thus prevent contamination of groundwater or other adjacent ecosystems.     

Greenhouse gases emission from soil contaminated with automobile industry residue in Brazil

Solid waste of the automobile industry containing large amounts of heavy metals might affect the emission of greenhouse gases (GHG) when applied to the soil. Accumulation of inorganic chemical elements in the environment generally occurs due to human activity (industry, agriculture, mining and waste landfills). Residues from human activities may release heavy metals to the soil solution, causing toxicity to plants and other soil organisms. Heavy metals may also be adsorbed to clay minerals and/or complexed by the soil organic matter, becoming a potential source of pollutants. Not much is known about the behavior of solid wastes in tropical soil as regarded as source of greenhouse gases (GHG). The emission of GHG (CO₂, CH₄ and N₂O) was evaluated in incubated soil samples collected in an area contaminated with a solid residue from an automobile industry. Samples were randomly collected at 0 to 0.2 m (a mix of soil and residue), 0.2 to 0.4 m (only residue) and 0.4 to 0.6 m (only soil). A contiguous uncontaminated area, cultivated with sugarcane, was also sampled following the same protocol. Canonical Discriminant Analysis and Principal Component Analysis were applied to the data to evaluate the GHG emission rates. Emission rates of GHG were greater in the samples from the contaminated than the sugarcane area, particularly high during the first days of incubation. CO₂ emissions were greater in samples collected at the upper layer for both areas, while CH₄ and N₂O emissions were similar in all samples. The emission rates of CH₄ were the most efficient variables to differentiate contaminated and uncontaminated areas.     

Isolation and Molecular Characterization of Heavy Metal-Resistant Azotobacter chroococcum from Agricultural Soil and Their Potential Application in Bioremediation

Pollution of soil with heavy metals, herbicides, antibiotics and other chemicals is known to have a negative effect on microbial activities. Therefore, the aim of this study was to isolate cultures of Azotobacter sp. from polluted and unpolluted soils and to study the effect of these pollutants on their growth. A total of 120 Azotobacter sp. were isolated from soils irrigated with wastewater (contaminated soils) and groundwater (uncontaminated soils). These isolates were screened for resistance to heavy metals, herbicide and antibiotics. Also, the soils from which the cultures were isolated were analyzed for the concentrations of Zn²⁺, Cd²⁺, Cu²⁺, Pb²⁺ and Mn²⁺ they contained. Contaminated soil showed high levels of heavy metals as compared to uncontaminated soil. The size of the Azotobacter population in contaminated soil was lower than that in uncontaminated soil. Of the Azotobacter isolates, 64 that were recovered from contaminated soil exhibited high resistance to heavy metals (Hg²⁺, Cd²⁺, Cu²⁺, Cr³⁺, Co²⁺, Ni²⁺, Zn²⁺ and Pb²⁺) and herbicide 2,4-D compared to 56 isolates from uncontaminated soil. Also, isolates from contaminated soil showed high resistance to chloramphenicol, nitrofurantoin and co-trimoxazole compared to those isolated from uncontaminated soil. The majority of Azotobacter isolates from contaminated soil showed multiple-resistance to different metal ions and antibiotics. All isolates failed to grow at pH less than 6. Salt concentration (5%) was found to be inhibitory to all isolates. The most potent isolates from contaminated soil that showed multiresistance to all substances tested were identified on the basis of morphological and biochemical characteristics, and 16S rRNA as A. chroococcum. These resistant isolates could be employed in contaminated soils and/or bioremediation.     

Appropriate experimental design for transplanting mussels (Mytilus sp.) in analyses of environmental stress: an example in Sydney Harbour (Australia)

Some locations in Sydney Harbour (Sydney, Australia) contain large amounts of contaminants (heavy metals and hydrocarbons), sometimes in concentrations thought to affect biological systems. In order to estimate effects of sediment-bound contaminants on the physiology of organisms living above the sediment, the rates of clearance and respiration and the efficiency of absorption of mussels, Mytilus sp., living in a contaminated location were measured, the scope for growth (SfG) was calculated and compared to that of mussels living in uncontaminated locations. Two different models were proposed to explain expected differences. The first was that the contaminants at the impacted location reduced the SfG of local mussels; the second was that at the contaminated location only those mussels survived that had a small SfG (genetic differences between populations might be a reason for differential survival). To test which model applied, mussels were transplanted between the contaminated and uncontaminated locations. Moving and disturbing mussels (handling, tagging and caging) required the inclusion of two control treatments. These treatments were essential for a proper evaluation of the results but have generally not been included in similar studies. Effects of moving were estimated by translocating mussels from the uncontaminated and from the contaminated location to similar locations. To estimate effects of disturbance, mussels in the experimental locations were given the same treatment as the experimental mussels, but were returned to the place of origin. It was predicted that translocating and disturbing mussels would have no effect on the SfG, which would be similar to that of mussels at the place of origin. As expected, SfG was smaller (because rate of clearance was reduced) in undisturbed mussels at the contaminated location compared with those of mussels at the uncontaminated location. Because there were significant effects of disturbance on the SfG of mussels at the contaminated location, it was concluded that this difference was not caused by differences between the amounts of contaminants in the two locations, but was caused by other confounding factors (physical disturbance by crabs and fouling organisms). That the interpretation of the results would have been different if proper controls were not included is discussed in this paper and the importance of proper experimental controls is stressed.     

The effect of endophytic Peyronellaea from heavy metal-contaminated and uncontaminated sites on maize growth,heavy metal absorption and accumulation

Heavy metal (HM) tolerance, effects on maize growth, heavy metal absorption and accumulation by endophytic Peyronellaea from HM-contaminated and uncontaminated sites were studied to evaluate the hypothesis that endophytes from HM-contaminated sites would enhance HM-tolerance in hosts. Although we found that certain endophytes improved tolerance of plants to heavy metals, isolates from the HM-contaminated site were not more tolerant to heavy metals than those from the uncontaminated site. Pot experiments indicated that growth and heavy metal absorption and accumulation by host plants in HM-polluted environments could be affected by inoculation with HM-tolerant endophytic fungi, and isolates showed a high intraspecific variability. However, there was no significant difference in growth between the maize inoculated with the endophytes from the HM-contaminated site and uncontaminated site under lead stress. Similarly, the HM content in the shoots and roots of maize inoculated with the isolates from the HM-contaminated site was not always higher than that in maize inoculated with endophytes from the uncontaminated site. Therefore, based on our experiments it is suggested that HM-tolerance due to endophytes and their effects on host plant growth and heavy metal absorption and accumulation were not correlated with origin of the endophytes.     

Metals in the tissues of the starry flounderPleuronectes stellatus from Nyiskii Bay (Southeastern Sakhalin)

We examined the concentrations of the heavy metals Zn, Cu, Cr, Pb, and Cd in the liver, gonads, and muscles of male and female starry flounderPleuronectes stellatus from Nyiskii Bay (off the northeastern coast of Sakhalin) in August 1996. It is shown that the accumulation of the metals was different in the organs and tissues of males and females. The greatest differences were recorded in the gonads. The concentrations of Zn and Cu were greater in the gonads of females, whereas the concentrations of Cd, Pb, and Cr were greater in male gonads. More Cd and Pb accumulated in the liver of females, whereas the concentrations of Zn and Cd were greater in the muscles of males. The results obtained seem to reflect the specific features of physiological processes in specimens of different sexes and the biochemical composition of the tissues of fish organs during the period of foraging and gonad development. The concentrations of metals in the muscles of starry flounder from Nyiskii Bay did not exceed the sanitary standards.     

The effects of 17β-oestradiol, testosterone and tamoxifen on the development of papillomata in Catostomus commersoni

Testosterone induced papillomata in 85% (11/13) of initially non-papillomatous white suckers Catostomus commersoni and increased papillomata growth in 100% (16/16) of initially papillomatous suckers. 17β-oestradiol induced papillomata in 83% (10/12) of initially nonpapillomatous suckers and increased papillomata growth in 100% (16/16) of initially papillomatous suckers. Less than 29% (4/14) of white suckers injected with tamoxifen developed papillomata, while complete papillomata regression was observed in 71% (10/14) of initially papillomatous suckers. In control groups 59% (27/46) of suckers either retained or developed papillomata and 27% (6/24) of suckers exhibited tumour regression. Protein kinase C (PKC) activity was significantly depressed and ornithine decarboxylase (ODC) activity was significantly elevated in steroid-treated papillomata v. normal lip epidermis. ODC activity was significantly depressed in tamoxifen-injected, regressing papillomata. There were no significant differences in plasma oestrogen and testosterone levels between papillomatous and nonpapillomatous female fish from a site contaminated with persistent organic chemicals and an uncontaminated reference site. Similarly, no significant differences in testosterone and 11-ketotestosterone levels were observed between papillomatous and non-papillomatous male fish.     

Root development of the Zinc-hyperaccumulator plant Thlaspi caerulescens as affected by metal origin,content and localization in soil

Penetration into and exploitation of contaminated soils by roots of hyperaccumulator plants is a prerequisite for efficient removal of heavy metals, i.e. efficacy of phytoextraction. This work was undertaken to study the development of roots of the Zn-hyperaccumulator Thlaspi caerulescens under various conditions of soil contamination. Rhizoboxes were constructed with a removable plastic front cover, and filled with soils containing different amounts and forms of metals (Zn, Cd and Pb). Treatments were: homogeneous soil profile, superposition of three layers, inclusion of contaminated soil into uncontaminated soil, or inclusion of uncontaminated soil into uniformly contaminated soil. Four seedlings were transplanted into each rhizobox, and development of the root system was periodically recorded for 133 days. At harvest, the biomass and size of the rosette of aerial parts were determined. The aerial biomass/root length fraction as well as the kinetics of root development varied according to the presence and localization of Zn. The distribution and morphology of roots at harvest were strongly dependent upon the metal content and form in soil. Roots exhibited a high affinity for the Zn-contaminated patches and showed two distinct morphologies according to the concentration of Zn in soil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Distribution and Risk Assessment of Heavy Metals in the Xinzhuangzi Reclamation Soil from the Huainan Coal Mining Area,China

The contamination of coal-mine soil by heavy metals is a widespread problem. This study analyzes the heavy metals (Cu, Zn, Ni, Pb, Cr, Cd, and Hg) found in 33 surface soil samples from Xinzhuangzi, China restored coal-mining land used as cultivated land. The results show that the selected elements were cumulative, especially for Cd. An index of geo-accumulation indicates that the soil was practically uncontaminated by Cu, Zn, Ni, and Hg, uncontaminated to moderately contaminated by Pb and Cr, and moderately to heavily contaminated by Cd. Based on the U.S. Environmental Protection Agency's ecological soil screening levels (Eco-SSLs) for Cu, Zn, Ni, Pb, and Cd and the Dutch Target and Intervention Values for Cr and Hg, the plants and soil invertebrates were not likely greatly influenced by the selected metals. Although the Cd concentration was found to have no significant effect on plants and soil invertebrates, it is the only metal with a concentration significantly above that required by Chinese standards (HJ/T 332–2006) for edible agricultural products, indicating that Cd is the predominant factor that determines the use of the reclaimed coal-mining area for farmland. Thus, employing the reclaimed land as farmland may not be a good option.     

Fish assemblage–environment relationships suggest differential trophic responses to heavy metal contamination

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Ornithine decarboxylase antizyme in kidneys of male and female mice.

Antizyme, a protein inhibitor of ornithine decarboxylase (ODC), was shown to be induced in mouse kidney by repeated injection of putrescine. Antizyme was also present as a complex with ODC in the kidney of untreated mouse. The amount of the renal ODC-antizyme complex was 3-fold higher in male mice than in female mice. On the contrary, the proportion of ODC present as a complex with antizyme was 24-fold higher in females than in males, and the decay of renal ODC activity after cycloheximide treatment was about 5-fold more rapid in females than in males. Administration of testosterone to female mice, a procedure known to prolong the half-life of renal ODC, increased both ODC activity and the content of ODC-antizyme complex, but decreased the antizyme/ODC ratio in the kidney. These results are consistent with the previous observation in HTC cells that the decay rate of ODC activity in the presence of cycloheximide correlated well with the proportion of ODC present as a complex with antizyme, suggesting the ubiquitous role of antizyme in ODC degradation.     

Elevated mortality among birds in Chernobyl as judged from skewed age and sex ratios

Background

Radiation has negative effects on survival of animals including humans, although the generality of this claim is poorly documented under low-dose field conditions. Because females may suffer disproportionately from the effects of radiation on survival due to differences in sex roles during reproduction, radiation-induced mortality may result in male-skewed adult sex ratios.

Methodology/Principal Finding

We estimated the effects of low-dose radiation on adult survival rates in birds by determining age ratios of adults captured in mist nets during the breeding season in relation to background radiation levels around Chernobyl and in nearby uncontaminated control areas. Age ratios were skewed towards yearlings, especially in the most contaminated areas, implying that adult survival rates were reduced in contaminated areas, and that populations in such areas could only be maintained through immigration from nearby uncontaminated areas. Differential mortality in females resulted in a strongly male-skewed sex ratio in the most contaminated areas. In addition, males sang disproportionately commonly in the most contaminated areas where the sex ratio was male skewed presumably because males had difficulty finding and acquiring mates when females were rare. The results were not caused by permanent emigration by females from the most contaminated areas because none of the recaptured birds had changed breeding site, and the proportion of individuals with morphological abnormalities did not differ significantly between the sexes for areas with normal and higher levels of contamination.

Conclusions/Significance

These findings are consistent with the hypothesis that the adult survival rate of female birds is particularly susceptible to the effects of low-dose radiation, resulting in male skewed sex ratios at high levels of radiation. Such skewed age ratios towards yearlings in contaminated areas are consistent with the hypothesis that an area exceeding 30,000 km² in Chernobyl’s surroundings constitutes an ecological trap that causes dramatic excess mortality.     

Employment of associative bacteria for the inoculation of barley plants cultivated in soil contaminated with lead and cadmium

In laboratory experiments, the rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens 7, Agrobacterium radiobacter 10, and Flavobacterium sp. L30 were found to have a relatively high resistance to the toxic heavy metals lead and cadmium (except that strain L30 was found to be sensitive to Cd). When introduced by means of seed bacterization, the heavy metal-resistant strains actively colonized the rhizosphere of barley plants cultivated in uncontaminated and contaminated soils. In both pot and field experiments, seed bacterization improved the growth of barley plants and the uptake of nutrient elements from soil contaminated with Pb and Cd. The bacterization also prevented the accumulation of Pb and Cd in barley plants, thereby mitigating the toxic effect of these heavy metals on the plants.     

Kinship analysis of Pacific salmon: insights into mating, homing, and timing of reproduction

Multilocus microsatellite genotypes were used to infer kinship and relatedness in two species of Pacific salmon from three populations in Washington State. Even in the absence of direct genetic data from parents, clustering of individuals according to allele sharing and reconstruction of parental genotypes allowed resolution of full- and half-sib relationships among 135 chinook salmon (Oncorhynchus tshawytscha) sampled as preemergent juveniles from 14 redds in the Dungeness River. Inferred reproductive behaviors included single-pair matings, polyandry in which females mated with two to three males at a single redd, polygyny in which males mated with two females at different redds, use of two redds by a single female, and use of one redd site by two females. Greater average relatedness (rxy) in the upper reach of the Dungeness River implied within-reach homing of returning adults. In steelhead trout (O. mykiss), the frequency of related pairs (dyads) of mature individuals that migrated up Snow Creek less than a week apart was greater than expected for randomly chosen dyads, as was the frequency of steelhead dyads that were spawned on the same day in the Forks Creek hatchery. These results imply a heritable basis for upstream migration date and maturation date in steelhead trout.     

Metal accumulation and metallothionein in brown trout, Salmo trutta, from two Norwegian rivers differently contaminated with Cd, Cu and Zn

In this work we have studied the accumulation of heavy metals in two brown trout (Salmo trutta) populations in their natural environment and the participation of metal binding to metallothionein (MT) in this process. Cd, Cu and Zn concentrations, total MT (including Cu MT) and Cd/Zn MT were measured in the gills, liver and kidney of trout inhabiting two rivers, one Cu-contaminated and the other Cd/Zn-contaminated, located at Røros, Central Norway. In both populations, high levels of Cu were found in the liver, whereas Cd was accumulated in liver and particularly in the kidney. The proportions of Cd/Zn MT and Cu MT in liver and kidney, but not in gills, reflected the accumulated and the environmental concentrations of these metals. The total Cu MT concentrations in the investigated tissues, however, were highest in trout from the river with the lowest ambient Cu concentration. It is suggested that MTs are of less importance in Cu-acclimated trout. The data also suggest that acclimation to a Cu-rich environment involves reduced Cu accumulation or increased Cu elimination. In trout from the Cd-rich environment, this metal was mainly bound to MT, whereas in trout from the Cu-rich environment Cd was also associated with non-MT proteins. These findings emphasize the importance to determine both Cd/Zn MT and Cu MT levels, when the participation of this protein in metal handling in trout tissues is investigated.     

Changes in ornithine decarboxylase activity in response to temperature stress and stimulation of juvenile hormone in Anastrepha fraterculus (Diptera, Tephritidae)

Ornithine decarboxylase (ODC) activity was analyzed in Anastrepha fraterculus (Diptera) females (4 days old) submitted to temperature stress (6 degrees C and 20/6 degrees C) and the topical application of juvenile hormone (JH). ODC activity and ejaculatory apodeme measurements (length and width) were made in males (15 days old) after 6 degrees C stress. JH dose of 500 ng and incubation of 3, 7, and 18 h increased ODC activity. Females reared at 6 degrees C and 20/6 degrees C had higher ODC activity than those reared at 25 degrees C. The treatment of 6 degrees C and JH incubation for 1 h increased ODC activity when compared to 6 degrees C treatments only. However, the treatment of 20/6 degrees C only after 3 or 18 h of JH incubation resulted in higher ODC activity than controls (20/6 degrees C) or 20/6 degrees C plus 1 h of JH incubation. Males did not undergo differences in ODC activity when reared at 6 degrees C or 25 degrees C but the ejaculatory apodeme measurements was higher in those reared at 25 degrees C than in those reared at 6 degrees C. The results can be considered an adaptive process to environmental changes.