Response of ectomycorrhizal and other Pinus sylvestris root-associated fungi to the load of allochthonous material from a great cormorant colony.

Colonies of piscivorous birds nesting in forests bring in loads of allochthonous material in amounts significantly exceeding atmospheric depositions, eventually causing major ecosystem changes. We studied the effect of increasing ornithogenic impact on ectomycorrhizal (EMF) and other root-associated fungi in a Scots pine forest affected by a colony of great cormorants. We evaluated quantitative data of ectomycorrhiza (numbers of root tips and morphotypes) and identified fungal species from pine root tips and from sporocarps collected in three designated study zones: D (zone of active nesting), E (colony margin), and G (pristine forest). Species-bearing sporocarps were absent in zone D, and their number steadily decreased in zone E and was significantly lower than in zone G. Species communities and occurrence frequencies of individual species differed significantly between the zones. Environmental factors, both directly (nutrient addition) and indirectly (changes in vegetation cover), associated with bird activity have significantly influenced fungal communities. We hypothesize that the first signs of increased avian impact on root-associated fungi are the reduced diversity of sporocarps and the low presence and diversity of boletoid fungi in root tips.

Hair geochemical composition of children from Vilnius kindergartens as an indicator of environmental conditions.

The research is based on analysis data of Cr, Cu, Mn, Ni, V, Zn (metals) and S in the hair of 47 girls and 63 boys from eight Vilnius kindergartens and the distribution pattern of high metal concentrations and bioavailability in snow-cover dust, also dust samples from vents of characteristic pollution sources. The kindergartens were selected according to topsoil total contamination index and dust-related indices. Significantly higher Cu, Mn, Ni and Zn concentrations in the hair of girls (means are 1.1, 1.9, 1.3, 1.2 times higher) and the differences between hair of genders according to inter-element correlation and clustering were found. Analysis of Spearman correlation coefficients between metal concentrations in hair of each gender and dust metal concentrations or metal loading rates at their residence sites revealed that for Mn, Cu and Zn, they are insignificant, while for Cr, Ni, Pb and V, they are mainly significant positive (except V in female hair). The correlation of the contents of Cr, Ni and V in dust with respective concentrations in hair was more significant for boys (p < 0.001) than for girls. Only a few cases with a significant Cr, Ni, Cu, Pb and Zn increase were revealed in hair of children attending polluted kindergartens in comparison with control. It was concluded that relationship between metal concentrations in hair and dust-related indices is more expressed for children's residence sites than for their kindergarten sites. The gender-based grouping and site-by-site study design are recommended in the studies of reflection of environmental exposure in hair.

Impact of Edaphic and Climatic Factors on Thymus pulegioides Essential Oil Composition and Potential Prevalence of Chemotypes.

Intraspecific chemical polymorphism is characteristic of essential oil bearing Thymus pulegioides (Lamiaceae). Soil chemical composition and climatic conditions can influence not only quantitative and qualitative composition of essential oils, but also on prevalence of different chemotypes in space. The purpose of study was to determine the impact of edaphic and climatic factors on T. pulegioides essential oil composition and potential chemotypes prevalence. It were investigated 131 habitats of T. pulegioides in all ten climatic sub-districts of Lithuania. Essential oils were isolated by hydrodistillation and analysed by GC-FID and GC-MS. The content of humus was estimated by oxidation method, the content of mobile potassium (K2O) and mobile phosphorus (P2O5)-flame photometry using 0.2 M HCl solution, the soil pH-electrometrically and 15 elements of soil-by energy-dispersive X-ray fluorescence. The meteorological data (temperature, rainfall, photosynthetically active solar radiation and sunshine duration) were obtained from the meteorological bulletins. Results showed that humus in soil positively, manganese and cobalt-negatively influenced on the essential oil accumulation in T. pulegioides. Carvacrol was the most common compound in the essential oil of T. pulegioides; higher amounts of sulphur and mobile phosphorus, lower amounts of sodium in soil, higher rainfall in blooming period are favourable for this compound and for potential prevalence of carvacrol chemotype. Geraniol was frequent in the eastern and the central part of Lithuania, where the temperature in vegetation season is higher, and in soils characterised by higher and lower amounts of sodium and chlorine, respectively. Thymol, linalool and α-terpinyl acetate were rare. Edaphic and climatic conditions can differently influence on essential oil yield and chemical composition of species as well as on potential prevalence of different chemotypes of same species in space.

Impact evaluation of marking Salmo trutta with Alizarin Red S produced by different manufacturers.

Fish otolith marking with the alizarin dye is a commonly used tool in sustainable fishery management. However, the reported effects of this dye on fish health are rather controversial and are possibly linked to differences in the composition of different brands of Alizarin red S (ARS). Laboratory experiments designed to elucidate effects of different concentrations of theoretically the same ARS as indicated by the CAS (Chemical Abstracts Service) number on fish at different development stages were carried out. The acute toxicity of ARS to Salmo trutta was found to be concentration- and fish developmental stage-dependant. Our study results showed that S. trutta sensitivity to ARS varies depending on its developmental stages as follows: fry (50-days after hatching) > alevins (30-days after hatching) > alevins (1-day after hatching). One of the tested ARS brands (purchased from VWR International LLC (Matsonford Road, USA)) was found to be several times more toxic to fish than another (purchased from Sigma-Aldrich (St. Louis, USA)), although according to the certificates of analysis, the tested substances were identical. Survival and growth of the S. trutta fry, which was marked with different ARS brands and stocked in the same natural stream, was investigated for two consecutive years. The results obtained indicate remarkable differences (p < 0.05) in the effects produced by the tested ARS brands, thus confirming our laboratory findings. The performed elemental analysis of the tested ARS dyes revealed significant differences in chemical impurities that these dyes contain. This study has, for the first time, expressed concern about the probable long-term impact of some ARS brands on the marked fish and their potential to bias the results of the studies dealing with ARS-marked fish.

Cyto/genotoxicological evaluation of hot spots of soil pollution using Allium bioassays in relation to geochemistry.

Soil from industrial and landfill sites affected by anthropogenic activity was screened for implicit negative effects in an Allium test-system in relation to geochemistry. The concentrations of 15 elements were compared to the ecotoxicologically-based soil guideline values. Admitted geoindices were used to classify test-soils according to risk/hazard categories. Test-soils were screened for the possible deleterious effects in common onion (Allium cepa L.) by employing a test battery of cytogenetic bioassays (root growth inhibition, mitotic activity, frequency of chromosome aberrations and micronuclei, and cell death rate) complemented with two assays of molecular DNA markers, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR). Soil from industrial sites was more severely polluted and more cytotoxic for onions compared to soil from landfill sites. However, the cyto/genotoxic outcome of soil exposure in A. cepa was the same for all test-soils; the detrimental effects were observed in onions treated with every test-soil. Thus, test-soils could not be classified as non- and genotoxic, although certain of them had permissible contamination levels. The chromosome aberration frequency and cell death rates were consistent with the intensity of soil contamination, contrary to the micronuclei rate, which was independent of the soil risk/hazard level. Despite a relationship between risk (RI) and total soil contamination (Z) geoindices, both indices correlated with a different Allium cyto/genotoxicity endpoint, although the Z index was preferred over the RI index as being more informative in correlation analysis. Allium bioassays complemented each other by depicting different aspects of exposure to toxic substances, and determination of cyto/genotoxicity in a battery of different bioassays is important in the risk assessment of ecologically dangerous soils, and an application of a test battery is strongly advised.

Response of Tradescantia plants to oxidative stress induced by heavy metal pollution of soils from industrial areas.

Numerous investigations have demonstrated that even soil in which concentrations of individual elements do not exceed permissible limits can cause harmful effects in living organisms. In the present study, polluted-soil-induced oxidative stress was evaluated using Tradescantia clone 4430, which is widely used for genotoxicity evaluations, employing biochemical (superoxide dismutase (SOD), contents of ascorbic acid (AA), carotenoids (Car), hydrogen peroxide (H2O2), chlorophyll (Chl) a/b ratio), and molecular (RAPD and differential display (DD-PCR)) markers after long-term exposure. The activity (staining intensity) of SOD isoforms in Tradescantia leaves was higher in plants grown in all heavy-metal-polluted test soils compared to the control. No direct link between the soil pollution category and the contents of AA, Car, Chl a/b in Tradescantia leaves was revealed, but the concentration of H2O2 was shown to be a sensitive biochemical indicator that may appropriately reflect the soil contamination level. Both short-term (treatment of cuttings with H2O extracts of soil) and long-term (0.5 and 1.0 year) exposure increased MN frequencies, but the coincidence of the MN induction and the soil pollution level was observed only in some cases of long-term exposure. Soil (geno)toxin-induced polymorphism in the RAPD profile was determined with two primers in plants after long-term exposure to soils of an extremely hazard category. Transcript profiling of plants after long-term cultivation in test soils using DD-PCR showed that the majority of differentially expressed transcript-derived fragments (TDFs) were homologous to genes directly or indirectly participating in photosynthesis, the abiotic stress response, and signal transduction cascades.

Accumulation of chemical elements in yellow-necked mice under a colony of great cormorants.

This study represents the first investigation into the accumulation of chemical elements in small mammals inhabiting the territory of a great cormorant colony. Trapping was done in the Juodkrante great cormorant colony, one of the largest colonies in Europe. The accumulation of 20 chemical elements in the bodies (muscle and bones) of yellow-necked mice (Apodemus flavicollis) was investigated using the energy-dispersive x-ray fluorescence equipment Spectro Xepos HE. Two groups of positively inter-correlated chemical elements (Mg, Al, P, Ca and Al, S, Cl, K) were identified. The concentrations of five elements differed significantly between mice trapped in different zones of the colony with differing intensities of cormorant influence: the values of K and Cu in A. flavicollis increased in line with an increase in the influence of the cormorants, while the concentrations of Rb and Pb decreased. The concentrations of Mn differed between zones, but were not related to the intensity of bird influence. Differences in the concentration of Zn (ANOVA F = 24.38; p < 0.001), Fe (F = 4.60; p < 0.05) and Mo (F = 4.47; p < 0.05) were related to the gender factor, all concentrations being higher in females. The concentrations of Zn were age-dependent, being highest in adult individuals (21.7 ±â€¯4.5 µg g-1) and exceeding those in subadult (19.4 ±â€¯3.4 µg g-1) individuals or juveniles (16.7 ±â€¯1.3 µg g-1). In general, the concentrations of accumulated elements in A. flavicollis from the territory of the cormorant colony were lower than in rodents from industrially polluted sites.

Metal bioaccumulation and mutagenesis in a Tradescantia clone following long-term exposure to soils from urban industrial areas and closed landfills.

Soil mutagens, particularly metals, may persist long after the source of pollution has been removed, representing a hazard to plants, animals, and humans in or near contaminated areas. Often, due to urban growth, previous land uses may be forgotten and hazards overlooked. We exposed Tradescantia clone #4430 plants to soil from two industrial areas (with different former uses) and two urban waste landfills in the city of Vilnius, all of which were long disused. Two modes of exposure were used: long-term exposure of growing plants in test soils for 0.5 or 1.0y, and short-term exposure of cuttings to water and dimethyl sulfoxide (DMSO) soil extracts. An increased frequency of micronuclei (MN) was observed with both modes of exposure. The concentrations of 24 metals and other elements were analyzed in the test soils and in above-ground plant parts, under both exposure modes, and the concentration coefficients (Cc) for various elements, the total contamination index (Zs) for soils and plants, and the bioaccumulation factor (BAF) for plants were calculated. These measurements allow a comparison of the contamination levels of soils and plants with equalized values. Metal accumulation levels in plants and soils showed significant differences, providing a better understanding of the genotoxicity of soils from closed landfills and highlighting the need to determine the concentrations of metals and other genotoxicants in plants in relation to genotoxicity.

Use of Tradescantia clone 4430 for direct long-term soil mutagenicity studies.

Soil mutagens permanently and directly affect terrestrial plants, soil microorganisms and invertebrates and indirectly impact vertebrate and human populations. However, the dynamic response to soil mutagens under conditions of long-term exposure has been studied insufficiently. The clonal nature of Tradescantia #4430 and the well-developed associated test systems for evaluation of genotoxicity allow investigation of the permanent effects of direct exposure to the soil not only on plants of the same genotype, but even with the same individual plants, as was done in the present study. Twenty soil samples of different origin were studied in Tradescantia after 0.5, 1 and 2 years of direct exposure to the soil, in order to examine the induction of pink cells (PC), colorless cells (CC) and branched hairs (BH). Additionally, after 0.5 years, micronucleus (MN) formation was assessed, and after 1 year, the variation of random amplified polymorphic DNA (RAPD) markers was evaluated. Nine soil samples were of urban origin, and the others came from different soils from former military grounds. The results of the 0.5-year exposure were compared with the effects of aqueous/DMSO extracts tested in previous work. The long-term exposure period allowed not only a better differentiation of the soils within territories, but also permitted detection of differences between soils of urban and military origin. The results also depended on the test used to evaluate genotoxicity. The 'burst' phenomenon, consisting of a sudden increase of BH variation and, especially, of CC, was observed only in tests of samples from several military sites, mainly after a 2-year exposure. MN formation was less frequent after 0.5 years of exposure compared with the results from aqueous extracts, and this difference was highly significant. The variation of the RAPD markers in a significant portion of the soil samples coincided with a higher level of variation according to other tests (PC, CC, BH), but predicting the genotoxicity of the soils based on their elemental soil composition at the end of the experiment was impossible. At several points where a 'burst' was observed, the concentration of a number of elements was lower. A cumulative effect is hypothesized to explain these observations. Tradescantia clone #4430 is the most suitable system for such studies.