The aim of the study was to determine the effects of various petroleum-derived substances (PDSs), namely petrol, diesel fuel, and spent engine oil, on life history traits of the bird cherry-oat aphid Rhopalosiphum padi L., and on the growth and chemical composition of its host plant—winter wheat Triticum aestivum L. Each substance was tested separately, using two concentrations (9 and 18 g kg−1 ). Plants were cultivated in both control and contaminated soils. In early October 2013, soil was contaminated and after 1 week, winter wheat seeds, ‘Batuta’ cultivar, were sown. In early June 2014, observations of the effect of petroleum-derived substances on traits of three successive generations of aphids were conducted. Aphids were inoculated separately on leaves using cylindrical cages hermetically closed on both sides. Contamination of aphid occurred through its host plant. Results showed that all of the applied petroleum-derived substances have a generally adverse effect on the developmental parameters in aphids, resulting in the decrease of its fecundity, shortening its average life span, and most often lowering of the population intrinsic growth rate. PDSs caused the limitation of growth in wheat plants; whereas, changes in nutrient contents and heavy metals depended on the part of the plant analysed, the substance applied, and on its dose. The negative relationships between the contents of both some macro-elements (Ca, K, P) and heavy metals (Mn, Cd, Cu, and Zn) and the developmental parameters of particular generations of R. padi were observed. The high susceptibility of R. padi to the presence of PDSs in the substrate for the host plant should be emphasised—the clear-cut changes in the life span and fecundity, with relatively small changes in the chemical composition of the plant, constitute an evident indication that the developmental parameters of aphids have the potential for the use as bio-indicator to evaluate the state of the environment contaminated by PDSs. | Petroleum-derived substances RhopalosiphumpadiL., Winterwheat, Heavymetals, Macronutrients, Micronutrients | 30 | 27000-27012 | 27

In vitro techniques may provide a suitable tool for effective propagation and conservation of plant species representing various ecological niches. The elaboration of such protocols is also prerequisite for selection of heavy-metal-tolerant plant material that could be afterwards used for restoration or remediation of polluted sites. In this study, culture protocol for Gypsophila fastigiata propagation was developed. The highest multiplication coefficient, which reached 6.5, and the best growth parameters were obtained on modified MS medium supplemented with 1.0 mg L−1 2iP and 0.2 mg L−1 IAA. The obtained cultures were treated with different concentrations of lead nitrate (0.1, 0.5, and 1.0 mM Pb(NO3)2) or cadmium chloride (0.5, 2.5, and 5.0 μM CdCl2). The growth parameters, photosynthetic pigments, and phenolic compound content were examined in order to evaluate whether tested metal salts can have an adverse impact on studied culture. It was ascertained that Pb ions induced growth disturbances and contributed to shoot wither. On the contrary, the proliferative shoot cultures were established on media containing Cd ions and the multiplication coefficients and shoot length increased on all media enriched with CdCl2. Chlorophylls and carotenoid contents were negatively affected by application of 5.0 μM of cadmium; nevertheless, in shoots treated with 2.5 μM CdCl2, increased accumulation of photosynthetic pigments occurred and their amount was similar to untreated culture. Adaptation to Cd was associated with stimulation of phenolic compound synthesis. Hence, we have reported on unambiguous positive result of in vitro selection procedure to obtain vigorous shoot culture tolerant to cadmium. | cadmium, facultative metallophyte, in vitro selection, lead, medium supplementation | 25 | 1-13 | 42

The paper presents an assessment of vegetation (composition and cover-abundance), nutrient supply, and especially sulfur accumulation in the trees foliage (birch and pine) used in reforestation and wood small-reed (Calamagrostis epigejos (L.) Roth) appearing in succession on reclaimed areas of the former Jeziórko sulfur mine (southern Poland, Tarnobrzeg region). In researched area, three categories of vegetation cover were determined: category D-degraded and unsuccessfully reforested plots, and two categories with successful reforestation: P-pine and B-birch stands. On each category, four study plots (4-6 areas each, depends on site category) were established. Soil and vegetation samplings (current year and 2-year-old pine needles, birch leaves, and wood small-reed foliage) were collected on the subplots established in regular grid square (10 × 10 m) in each category. Basic soil properties and nutrient content in soils and vegetation were analyzed. Trees grew well in areas where neutralization and reclamation treatments were carried out properly and showed a good supply of nutrients (exception of phosphorus and nitrogen), while on category D, only herbaceous vegetation with low cover-abundance and dominated by wood small-reed were noted. Linear correlations between the soil and trees nutrients content occurred, while the correlations between the soil and wood small-reed did not occur. Wood small-reed did not display increased sulfur uptake which may indicate a strategy of blocking pollutant uptake from the soil and may be recommended as a species resistant to sulfurous soils. | Frasch method; Plant nutrition; Reclamation; Reforestation; Sulfur contamination; Sulfur extraction by underground melting method | 30 | 20556-20566 | 24

The aim of this work was to analyse the response of Rosa rubiginosa to salinity induced by different concentrations of sodium chloride and calcium chloride (0, 25, 50, 100, 150 and 200 mM). Besides salt accumulation and pH changes, other parameters were investigated including photosynthetic activity, leaf water content, the dynamics of necrosis and chlorosis appearance and leaf drying. The study was complemented with microscopic analysis of changes in leaf anatomy. R. rubiginosa was more sensitive to the salinity induced by calcium chloride than by sodium chloride. Plant response to salinity differed depending of the salt concentration. These differences were manifested by higher dynamics of necrosis and chlorosis appearance and leaf drying. CaCl2 showed greater inhibition of the photosynthetic apparatus and photosynthetic activity. Treatment with CaCl2 caused more visible deformation of palisade cells, reduction in their density and overall reduction in leaf thickness. The study demonstrated higher accumulation of CaCl2 in the soil, and thus greater limitations in water availability resulting in reduced leaf water content and quicker drying of leaves as compared with NaCl-treated plants. | Salinity, Chlorosis, Chlorophyll fluorescence, Photosynthesis, Leaf anatomy | 25 | 1-11 | 228

We studied the bioaccumulation of lead in selected tissues of Prussian carp Carassius gibelio (Bloch, 1782) during 12 and 24 months exposure to different doses of this metal in feed and the elimination of lead from tissues during the following 12-month depuration period. Lead concentration was determined using atomic absorption spectrometry method. The highest lead concentrations were observed at 2.0 ± 0.54 to 7.4 ± 1.1 mg kg(-1) in the kidney, 3.0 ± 0.13 to 5.2 ± 0.17 mg kg(-1) in the bone, and 4.5 (±0.4) mg kg(-1) in the hepatopancreas of fish from groups exposed to lead dietary concentration from 8 to 49 mg kg(-1) for 24 months. The rate of accumulation were generally the highest at the beginning of exposure as evidenced by the highest monthly increments of bioaccumulation observed after 3 months of contamination for muscles, hepatopancreatic gland, intestine, and gills. Also analysis of the monthly increments of lead bioaccumulation in bone tissue and the highly significant coefficients of correlation indicate that the dynamics of accumulation are clearly dependent on dose of exposure. Depuration of accumulated lead from the organs depended mainly on tissue and duration of elimination period. Very rapid depuration was observed in soft tissues such as the intestine or muscles. Very low elimination was observed for scales and bones where until the end of the experiment highly significant lead concentration differences were observed in all groups in relation to the control group. Chronic dietary exposure in the range of 8-49 mg Pb kg(-1) resulted in no significant effects on the growth and survival of Prussian carp females. | Prussian carp, dietary, lead, exposure, accumulation, depuration | 30 | 3122-3132 | 20

In ants, intra and inter-colony variation in body size can be considerable, even in monomorphic species. It has been previously shown that size-related parameters can be environmentally sensitive. The shape of the body size distribution curve is, however, rarely investigated. In this study, we measured head widthes of the black garden ant Lasius niger workers using digital methods. The ants were sampled from 51 colonies originating from 19 sites located along a metal pollution gradient, established in a former mining area in Poland. Total zinc concentrations in random samples of small invertebrates were used as a measure of site pollution levels. We found that the skewness of head size distribution grows significantly in line with the pollution level of the site, ranging from values slightly below zero (about -0.5) in the least polluted site up to a positive value (about 1.5) in the most polluted site. This result indicates that the frequency of small ants grows as pollution levels increase. The coefficient of variation, as well as the measures of central tendency, was not related to the pollution level. Four hypotheses explaining the obtained results are proposed. The bias towards the higher frequency of small workers may result from energy limitation and/or metal toxicity, but may also have an adaptive function. | ants, skewness, monomorphism, body size, metal, pollution | 30 | 6126-6134 | 22

Lead is reported to be an endocrine disruptor. In the current study, we exposed female Prussian carp to artificial feed, supplemented with five doses of dietary Pb (0, 1 (control), 8, 13, 24, and 49 mg/kg) over either a 24-month period or a 12-month period, followed by further 12 months where the fish were fed the control diet. Periodically, oocyte maturation, gonadosomatic index (GSI), ability to secret luteinizing hormone (LH) as well as gonad Pb concentrations were measured. It was found that the reproductive system of the female Prussian carp is not indifferent to chronic exposure to lead. The negative effect was manifested by a decrease in the GSI after 12 months despite the fact that a higher proportion of oocytes at more advanced maturity stages were concurrently observed. After 12 months of exposure, the effect on LH secretion varied according to the dose. In the group exposed to the lowest dose (8 mg/kg), LH decreased spontaneously, and in the groups exposed to the highest two doses (24 and 48 mg/kg), a significantly higher LHRH-A-stimulated LH secretion was observed. After 24 months of lead exposure, the effects on oocyte maturation and size and on GSI values were not pronounced. Analysis of the effect of lead exposure on LH secretion showed that the relationships were similar to those observed after 12 months but nonsignificant. During chronic a 24-month period exposure to Pb, Prussian carp female appears to acclimate to Pb doses used in the experiment. After 12 months of exposure and 12 months of depuration, the levels of spontaneous and stimulated LH secretion observed in all the groups were similar to the control, which is evidence that depuration eliminates the previously observed effects of exposure to lead. Lead is easily accumulated in the ovary, reaching a fivefold higher level (0.8 mg/kg tissue) compared to the control (0.15 mg/kg tissue), but after discontinuation of exposure, this organ is quickly depurated. The results indicate that environmental Pb can be a potent endocrine disruptor affecting ovarian steroidogenesis, gametogenesis, and ovulation, which may lead to adverse impacts on fish reproduction and population density and that female Prussian carp become resistant to the negative effects of lead with advancing age, and their organs cope by reaching a state of homeostasis. | Lead, Prussian carp, Oocyte maturation, Luteinizing hormone secretion, Bioaccumulation, Depuration | 30 | 2465-2478 | 4

The aim of this research was to modify the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method for the determination of organochlorine and organophosphate pesticides in fatty animal matrices such as fish muscle tissues of carp and sturgeon collected from Carp Valley, Lesser Poland. Pesticides extraction effectiveness was evaluated at 0.030 mg kg−1 spiking level and efficiency of the dispersive-solid-phase extraction (d-SPE) clean-up step was evaluated by comparison testing two different d-SPE clean-up stages, first the addition of the d-SPE sorbent combination (PSA + SAX + NH2), and secondly the addition of C18 after extracts enrichment with the d-SPE sorbent combination (PSA + SAX + NH2), introducing a novel concept of clean-up named dual-d-SPE clean-up. Analysis of pesticide residues was performed by Gas Chromatography Quadrupole Mass Spectrometry (GC/Q-MS) working in selected-ion monitoring (SIM) mode. Linear relation was observed from 0 to 200 ng mL−1 and determination coefficient R 2  > 0.997 in all instances for all target analytes. Better recoveries and cleanliness of extracts in both samples, carp and sturgeon tissues, were obtained after C18 addition during the dual-d-SPE clean-up step. Recoveries were in the range 70–120 %, with relative standard deviation lower than 10 % at 0.030 mg kg−1 spiking level for most pesticides. LODs ranged 0.001–0.003 mg kg−1, while LOQs ranged 0.004–0.009 mg kg−1. The proposed method was successfully applied analyzing pesticide residues in real carp and sturgeon muscle samples; detectable pesticide residues were observed, but in all of the cases contamination level was lower than the default maximum residue levels (MRLs) set by the European Union (EU), Regulation (EC) N 396/2005. | pesticides, GC-MS, QuEChERS, d-SPE, fish tissue, fatty animal matrix | 30 | 369-378 | 1

The aim of this work was the evaluation of QuEChERS extraction method for polycyclic aromatic hydrocarbon (PAH) determination in various types of tea. In the experiment, different kinds of extraction solvents, sorbents and a final method of sample preparation were compared. The final extracts were analysed by gas chromatography-selected ion monitoring-mass spectrometry. The results suggest that acetonitrile extraction, clean up with SAX and final liquid–liquid extraction was the best combination giving the most purified extracts and acceptable compound recoveries for different types of teas. In the study of real samples, compounds belonging to light PAHs were mostly detected, and heavy polycyclic aromatic hydrocarbons, including benzo[a]pyrene, were not identified in any of samples. | GC-MS, Extraction, PAHs, Tea, QuEChERS, SPE sorbents | 30 | 1326-1338 | 2

The purpose of the present experiment was to describe the influence of various mineral fertilizers on the development of Larix decidua Mill. seedlings subject to the influence of simulated acid rain of pH 2.5, 3.0, 3.5, 4.0, 4.5. The seedlings were fertilized every month (from April up to October), in order to counteract acid rain. The composition of the fertilizers was diversified in order to select an optimum variant. Fertilizer with NK applied to the needles (Florovit) and NPK applied to the soil (Fruktus 2) as well as dolomite lime applied to the soil. One row per set was left without fertilization as a control for each pH step of acid rain treatment. The analysis of biometric characteristics showed that a two-year period of investigations was too short to estimate effects on Larix decidua Mill. seedlings. It, however, indicated important trends. High correlations were found between particular growth parameters i.e. diameter of root neck, root mass, total mass of: seedlings, needles, main shoots, lateral shoots, ligneous parts, total overground parts. No strong correlations were observed between the above mentioned parameters and the total height of the seedlings. Among the different treatments the best development was achieved at pH 4.0, whereas the weakest development was observed at pH 2.5. The analysis showed a decrease in biometrical features along with an increase in acidity of acid rain solution. The first year of the experiment had the greatest influence upon the growth of Larix decidua Mill. seedlings. Significant improvement in growth, in relation to control plots, was achieved by using NK-fertilizers onto needles — the best results, and NPK onto soil. The best increment of the height of seedling was noticed at pH 3.5. In the second year of the experiment the effects of pH of acid rain and fertilization decreased — no significant effect on biometrical features was observed. Applaying NPK fertilizer onto soil showed a positive effect in relation to dolomite treatment. The best increment was noticed at pH 4.0. | 0 | 93-107 | 1-2