The roles of ectomycorrhizal fungi and bacteria associated with corresponding fungal species in distribution of heavy metals within roots and shoots of inoculated pine (Pinus sylvestris L.) seedlings were determined in this study. The mycorrhizal fungi forming different morphotypes were identified by PCR-RFLP using respective primers for an internal spacer transcribed region (ITS) of fungal rDNA. Amongst five fungal species detected, three were identified as Scleroderma citrinum, Amanita muscaria and Lactarius rufus. These fungi used for inoculation of pine seedlings significantly reduced translocation of Zn(II), Cd(II) or Pb(II) from roots to shoots, and the pattern of metal-accumulation was dependent on the fungal species. Ectomycorrhizae-associated bacteria identified as Pseudomonas were used as an additional component of the pine inoculation. These dual root inoculations resulted in higher accumulation of the metals, especially Zn(II), in the roots compared to the inoculation with fungal species alone. Consequently, dual inoculation of pine seedlings could be a suitable approach for plant protection against heavy metals and successful planting of metal-polluted soils.

This paper reports the effect of using coarse substrates in the upper layer of a gravel-based tidal flow reed bed treating pig farm wastewater. The aim of this unconventional medium arrangement is to seek a solution for the problem of clogging that frequently takes place during the treatment of strong wastewaters. Results from lab-scale experiments demonstrated that, in general, employing coarse substrates in the upper layer of the reed bed gave greater efficiency for the removal of organic matter and suspended solids, due to reduced clogging. A specific clogging tendency rate was defined to quantitatively describe the clogging behaviour. Calculation of the tendency rate revealed that the unconventional medium arrangement had a clear advantage over the conventional arrangement of employing fine gravel or sand in top layer.

Wastewater treatment plants (WWTP) with insufficient technologies for wastewater purification often cause a distinct nutrient pollution in the receiving streams. The increased concentrations of dissolved nutrients can severely disturb the ecological integrity of streams, which has been recently shown for basic ecosystem processes like mineralization of coarse particulate organic matter (CPOM). The present study investigated the impact of a modern WWTP (Zentralkläranlage Jena) on breakdown rates of CPOM exposed in net bags (1 mm mesh size) to the effluent of a large municipal WWTP and an upstream control site in the Saale River (Thuringia, Germany) from April to October 2005. Control and effluent site differed significantly in water chemistry with increased concentrations of dissolved organic carbon (DOC), ammonium, sulfate, and chloride at the effluent site, while the control site displayed higher concentrations of nitrate. However, breakdown rates of toothpickers and small twigs were not significantly different between the sites, whereas breakdown rate of leaf litter was significantly higher at the effluent site (k = 0.0124 day⁻¹) than at the control site (k = 0.0095 day⁻¹). Benthic invertebrate assemblages inhabiting the sandy stream bed at both sites were dominated by Chironomidae and Oligochaeta, typical inhabitants of fine sediments. Although the Shannon diversity of the benthic invertebrates was slightly higher at the effluent site (0.85) than at the control site (0.63), no significant difference could be detected. Bacterial numbers in water samples and surface biofilms on glass slides also displayed no significant differences between the two sites. This study showed that the effluents of a WWTP with modern technologies for wastewater purification did not directly affect breakdown rates of CPOM, bacteria numbers in epibenthic biofilms and the water column, and the community composition of sediment inhabiting aquatic macroinvertebrates in an effluent-receiving river with already increased concentrations of dissolved nutrients.

A review is given in this paper of the up-to-date results observed in differentiation and transformation studies on petroleum-type pollutants in underground and surface waters. Water and particulate matter derived from the locality of Pančevo Petroleum Refinery, Serbia (River Danube alluvial formations). It was shown that distributions of n-alkanes, steranes and triterpanes, and δ¹³CPDB values of n-alkanes may successfully be used for qualitatively differentiating the petroleum-type pollutants from native organic matter in recent sedimentary formations. In underground waters, a petroleum-type pollutant is exposed to microbiological degradation which is manifested through relatively fast degradation of n-alkanes. Following an almost complete degradation of crude oil n-alkanes in underground water, the biosynthesis of novel, even carbon-number C₁₆-C₃₀ n-alkanes may be observed. It is shown that the n-alkane distribution observed in a petroleum-type pollutant may depend on the intensity of its previous interaction with water. The fate of petroleum-type pollutants in environmental waters may be predicted through laboratory simulative microbiological degradation experiments by using microorganism consortiums similar to those observed under relevant natural conditions, as well as on corresponding nutrient base.

In order to achieve remediation of contaminated substrates, phyto-extraction in pot experiments utilizing lettuce seedlings as universal accumulator plants was investigated. As test substrates, mine tailings from Shiheung and Okdong mines in Korea (particularly high in Pb, Zn, Cu, and Cd), as well as samples from historic mining site at Oberzeiring in Austria (particularly high in Pb, Sb and As) were used, and compared with adjacent farmland soils. After 21 days of growth in the test substrate, the lettuce plants were harvested, and the adjacent soils parted in bulk and root soils. Special soil bacteria, adapted to high Cd levels (Exiguobacter sp.) and capable of adsorbing large amounts of cadmium from solution, as well as perlite (Samson Perlite Inc.) were added to the test substrates before plant growth. Speciation changes in the solids were investigated by sequential leaching, utilizing neutral MgCl₂ (exchangeable), 0.16 M acetic acid, hydroxylamine pH 2, oxalate pH 3, H₂O₂ oxidation, and reflux with aqua regia. Plant growth induced differentiation between root and bulk soils, the differences were more pronounced for the non-contaminated controls. The iron-hydroxide phase increased about 30%, and also the amount of iron-hydroxide bound Be, Cd, Co, Cu, Mg, Mo, Sb and V concentrations, coming mainly from less mobile fractions. The Mn hydroxide phase, however (hydroxylamine), remained rather constant. After plant growth, the root soils were significantly lower in available P, and significantly higher in available Ca, Mn, and Na than the corresponding bulk soils. Addition of Cd-adapted soil bacteria led to a severe decrease of plant yield, but metal uptake changed in both directions. Exchangeable P in both root and bulk soil decreased, and Be, Co, Cr, Fe, K, Li, Mg, Mn, Ni, and Sr in the residual organic fraction increased. This can be interpreted as competition for nutrients, dissolution of residuals by bacterial action, and adsorption to a tightly bound biomass. Addition of perlite hardly affected the plant yield, and again metal uptake changed in both directions, but led to a decrease of siderophilic elements in the Fe- and Mn hydroxides of the bulk soil. In the root soil, perlite addition above all decreased available K, P and As, with respect to the untreated samples. Bacteria addition to perlite treated soils shifted some elements from weak acid mobile towards less available fractions.

Constructed wetlands have been recognized as offering a removal treatment option for high concentrations removal of chemical and biological contaminants in domestic wastewater. The enteric protozoan parasite Cryptosporidium is considered one of the highly resistant to treatment and highly infectious organisms to humans and animals. Moreover, some species of Cryptosporidium are known to have a zoonotic nature. In this investigation a pilot scale for domestic wastewater treatment system was used, consisting of the following steps in series: (1) up-flow anaerobic sludge blanket (UASB) reactor, (2) free water surface (FWS) wetland unit, and (3) sub-surface flow (SSF) wetland unit. This treatment system was fed with domestic wastewater to assess its efficiency in removing Cryptosporidium oocysts. The obtained Cryptosporidium oocysts were detected and enumerated by two different staining techniques 'acid fast trichrome (AFT) and modified Ziehl Neelsen (MZN) stains'. Polymerase chain reaction (PCR) technique was also used to detect Cryptosporidium DNA in wastewater samples. Results revealed that anaerobic treatment (using UASB reactor) could remove about 53.1% of Cryptosporidium oocysts present in raw wastewater. The in-series connection between the two wetland units allowed complete elimination of Cryptosporidium oocysts as the first (FWS) wetland unit removed 95.9% of the oocysts present in anaerobically treated wastewater and the remaining portion of oocysts was completely removed by the second (SSF) wetland unit. Cryptosporidium oocysts were detected in 95.8% of raw wastewater samples with a mean count of 43.8 oocysts/l when AFT stain was used while they were detected in only 87.5% of raw wastewater samples with a mean count of 35.6 oocysts/l when MZN stain was used. Polymerase chain reaction (PCR) technique was able to detect Cryptosporidium DNA in only 45.8% of raw wastewater samples. Positive PCR results were only achieved in wastewater samples containing 52 oocysts or more per liter.

Increasing environmental concentrations of platinum group metals (PGMs), in particular platinum (Pt), rhodium (Rh) and palladium (Pd), from catalytic converters has been reported worldwide. The impact of these three metals on the uptake and use of essential mineral nutrients was examined using two plant models: the submerged aquatic plant, Elodea canadensis, and the terrestrial emergent plant, Peltandra virginica. Plants were grown for 2 weeks in nutrient solutions with either Pt⁴⁺ at concentrations between 0.05 and 5 mg/L, or a 0.1 mg/L Pt⁴⁺, Rh³⁺, Pd²⁺ mixture. Some treatments received additional Ca²⁺, Zn²⁺, or humic acid (with varying pH) to study how these conditions affected PGM uptake. Metal concentration analyses were conducted using a graphite furnace atomic absorption spectrometer (GFAAS) or an inductively coupled plasma emission spectrometer (ICP). Growth response was assessed through total chlorophyll content. There was significant Pt accumulation in plant tissues, from 55 to 326 times the concentration in nutrient solution. At pH 8, the addition of humic acid doubled Pt accumulation in comparison to the control. Additional exogenous minerals did not significantly affect PGM uptake, nor did the uptake of PGMs interfere with the uptake of Ca, Fe or Cu. Synthesis of chlorophyll in new shoots was not affected by Pt accumulation; however, visible chlorosis was observed in older shoots at 5 ppm Pt. Roadside Daucus carota samples from four heavy traffic locations in Dutchess County (New York) were also assessed for PGM content. Pt, Pd and Rh concentrations averaged 14.6, 10.2, and 0.7 μg/g, respectively.

According to recent insight, the toxicity of metals in soils is better related to the free metal ion (FMI) activity in the soil solution than to the total metal concentration in soil. However, the determination of FMI activities in soil solution is a difficult and time-consuming task. An alternative is to use empirical equations (so called transfer functions (TFs)) that relate FMI activity in solution to the reactive metal concentration in the solid phase and to soil properties (pH and organic matter content). Here we test the applicability of two sets of TF for Cd and Pb using independent data from a wide range of soil types and regions that are not represented in the datasets used to derive the TFs. From these soils, soil solution was extracted using four different methods. For all these extracts, FMI activities were calculated from total concentrations in solution using the speciation program WHAM VI. In some of the soils, Cd and Pb FMI activities were also measured using a Donnan membrane technique. Most of these FMI activities deviated from the TF predictions by less than one order of magnitude and were within the 95% confidence interval of the TFs, irrespective of the method used to extract soil solution. Predictability was higher for Pb than for Cd and differed also between the two TF sets.

Comparing today's atmospheric deposition records with the elemental concentration and the net-uptake rates of ombrotrophic Sphagnum mosses from eight German and Belgian peat bogs revealed that most of all the quality and number of regularly obtained deposition monitoring data is not satisfactory. Moreover, it seems likely that the deposition rate, determined by Sphagnum mosses, does not reliably reflect the record of the total open field deposition indicated by the deposition monitoring data. The moss data, too, show a distinct spatial variability possibly because the geochemistry of peat mosses differs according to the annual growth in height, the total surface area and the surface roughness of the receptor 'peat moss' (special interception deposition). Increased Ti concentration values, for example, combined with a high annual growth rate in height at the hollow moss S. cuspidatum resulted in generally high Ti net-uptake rates and a high Ti inventory (total Ti in sample). We, therefore, suggest that productive Sphagnum species might be able to fix more Ti particles on their larger surface area than less productive species do. Moreover, the results demonstrate that for reliably calculating Sphagnum elemental net-uptake rates, as well as for all quantification of Sphagnum or peat geochemistry on a time and area basis, an accurate knowledge of the period the collected samples were exposed to atmospheric deposition is required. In particular, to do reliable reconstructions of past atmospheric deposition rates using peat deposits, further studies are needed to precisely specify the spatial variability in the geochemistry of living Sphagnum mosses.

Sedimentprofiles of the last 4,000-14,000 years are presented from three dimictic lakes in Mecklenburg-Vorpommern (North-eastern Germany). Sedimentological composition, major trace elements and nutrients as well as parameter for core chronology (palynology, ¹⁴C-AMS) were investigated in order to reconstruct the historical development of the lakes during the Holocene. Palynological results reflect different human settlement phases and environmental changes from the late Pleistocene to the Subatlantic. Since the Middle Ages, a permanent settlement in the catchment area resulted in higher sedimentation rates in the three lakes. Variations in sediment composition like organic matter, carbonate and mineral content were caused by different land management techniques and natural changes in the catchment area. The phosphorus accumulation increased in the upper sediment layers, but the highest phosphorus accumulations were not found in the industrial phase, but in older sediments associated with human settlement activities in the catchment area. The heavy metals lead and zinc increase in the uppermost part of all three lakes reflecting the atmospheric anthropogenic input during the last 150-200 years.