by PSP Volume 20 No 11a Fresenius Environmental Bulletin

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1 PHOSPHOROUS AND CERTAIN METALS IN PARASOL MUSHROOMS (MACROLEPIOTA PROCERA) AND SOILS FROM THE AUGUSTOWSKA FOREST AND EŁK REGION IN NORTH-EASTERN POLAND Anna K. Kojta 1, *, Magdalena Gucia 1, Grażyna Jarzyńska 1, Marta Lewandowska 1, Anna Zakrzewska 1, Jerzy Falandysz 1 and Dan Zhang 2 1 Research Group of Environmental Chemistry, Ecotoxicology & Food Toxicology, Institute of Environmental Sciences & Public Health, University of Gdańsk, 19 Sobieskiego Str., Gdańsk, Poland 2 Key Laboratory of Mountain Environmental Diversity and Control, Institute of Mountain Hazards and Environment, Chinese Academy of Sciences and Ministry of Water Conservancy, Chengdu 641, China ABSRACT This report provides, using a validated analytical method, data on a baseline concentrations and bioconcentration potential of Ag, Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mn, Mg, Na, Ni, P, Pb, Rb, Sr and Zn by Parasol Mushroom collected from two spatially related sites - the stands in the Augustowska Forest and the stands in the outskirts of the town in north-eastern part of Poland. The total elements content was determined by ICP-AES and CV-AAS for Hg. Parasol Mushroom efficiently accumulated Ag, Hg, Cu and K in fruiting bodies and moderately of Ca, Cd, Mg, Na, P, Rb, Zn. Tasty caps of Parasol Mushroom from forested areas in north-eastern Poland are rich in essential elements such as Rb, K, P, Cu, Zn, Mg and Na. Nevertheless, a portion of specimens collected can contain in flesh Cd and Pb in concentrations exceeding tolerance limits set in the European Union for mushrooms. Eating these caps can result in an exceeding the safety intake margins of Hg (a slightly), and more of Cd, and especially, if eaten more than once per week. Measured elements concentration in soil samples confirmed that fruiting bodies were collected from unpolluted areas. The data obtained are consistent with the data resulting from the various studies carried out on unpolluted areas. Received data about elements concentration in both soil and flesh of the mushroom were statistically analyzed, including multifunction analysis (e.g. Mann-Whitney U test, CA, PCA). KEYWORDS: Field Parasol, food, heavy metals, ICP-AES, metals, mushrooms * Corresponding author 1. INTRODUCTION Parasol Mushroom (Macrolepiota procera) (Scop.: Fr.) Singer, choice edible mushroom in the genus Macrolepiota of the family Agaricace, is saprophytic species. As many other saprophytic but also mycorrhizal mushrooms take a vital part in biogeochemical turnover of metals, semimetals and other mineral elements absorbed from dead organic matter and soils for use by biosphere, which is a key role played by many fungi as decomposers in ecosystems [1]. An example could be silver that is abundant in fruiting bodies of some macromycetes and probably this biota play a possibly major role in Ag biogeochemical cycling in soils [2-5]. As a result of these, fungal fruiting bodies (carpophores) can be enriched in many metallic elements and also in metalloids, even if they grown-up at the background (uncontaminated) areas, while heavy contaminated when emerged at the polluted soils [6-11]. Intake rates by men of wild mushrooms seem to vary highly between the nations, regions of the world or the families and individuals, e.g. intake rates per capita shows values such as 1 kg (Sweden), up to 10 kg (Czech Republic) and kg (Liangshan Yi nationality in China) [12]. This report provides, using a validated analytical methods, data on a baseline concentrations and bioconcentration potential of 20 elements by Parasol Mushroom collected from two spatially related sites - the stands in the Augustowska Forest and the stands in the outskirts of the town in north-eastern region of Poland. This study is a part of ongoing survey aimed to investigate metallic elements and metalloids in wild grown mushrooms to understand their complex mineral constituents composition and interrelationships, bioconcentration and bioindication potential, impact of soil bedrock geochemical composition, contamination status as well as mineral constituents intake rates, nutritional significance and risk to mushroom consumers [8, 12-47]. 3044

2 2. MATERIALS AND METHODS Mature mushrooms were collected in the Augustowska Forest in Podlaskie Voivodeship in 2000 and outskirts of the town in Warmia and Mazury Voivodeship in The sampling sites are a typical forested and agricultural area not subjected for direct human activities. From the stands in the Augustowska Forest together with the Parasol Mushroom specimens were collected also the samples of forest soil upper horizon (0-10 cm, g, after removing superficial layer of litter and organic detritus). The Augustowska Forest is a large forested and lake area (1140 km 2 on country territory) in north-eastern corner of Poland. Unpolluted by industry environment is an unquestionable advantage of those sampling sites. Fresh fruiting bodies were cleaned from any present foreign materials using a plastic knife and divided into two morphological parts; caps and stipes. Each sample was dried at 65 ºC to constant weight in electrically heated laboratory drier. Next, the samples were pulverized in an agate mortar and in polyethylene bags in dry condition. Approximately 500 mg of homogenized fungal samples were weighted to closed polytetrafluoroethylene (PTFE) vessel and pre-digested (~24 h) using 7 ml of concentrated (20 %) nitric acid solution (Suprapure, Merck) at room temperature. Next, the vessels were closed and pressure/ temperature digested in an automatic microwave digestion system of type MARS 5 (CEM Corporation, Matthews, NC, USA). The soil samples were air dried in room temperature for several weeks and then sieved through a sifter (pore size 0.2 mm). Then, the substratum samples (~2.5 g) in quartz vessels were cold (room temperature) extracted with concentrated (20 %) nitric acid solution (Suprapure, Merck) for 24 h. The extracts were filtered using Whatman no. 42 filter paper before being diluted to 50 ml using deionized water [48, 49]. Precision and reproducibility of the methods were determined by analyzing blank samples, duplicates of selected samples and calculating the coefficient of variation, which were lower than 5 % for each element. Additionally, the methods of elements determination were validated and controlled by the analysis of several certified reference materials [10, 43, 50]. Concentrations of twenty chemical elements (except Hg) in fruiting bodies and soil samples extracts were determined by inductively coupled plasma-atomic emission spectrometry (Optima 2000, Perkin- Elmer, USA). Determination of mercury content was by cold-vapor atomic absorption spectrometry (CV-AAS) using an automated kept 3200 Mercury Monitor (Thermo Separation Products, USA). Details on used method of mercury determination were described in previous articles [51, 52]. The methods limits of detection for Ag, Al, Ba, Ca, Cd, Co, Cu, Cr, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Rb, Sr and Zn were between µg g -1 dw, and µg g -1 dw for Hg. Coefficients of variation for these measurements on routine runs were well below 10 %. The element concentrations in both soil and fungi specimens were compared using statistical methods, including multifunction analysis (e.g Mann-Whitney U-test, CA, PCA at significance level p<0.05). 3. RESULTS AND DISCUSION Many edible species of wild-grown mushrooms are food for numerous animals and are a popular foodstuff, often collected and cooked fresh by many people, but also available commercially as fresh, dried, pickled or as ingredients in other products. A need of knowledge on chemical composition of edible mushrooms as related to nutritional value and risk of hazardous compounds is undisputable. The fruiting bodies of mushrooms can be also considered as indices of environmental pollution, e.g. heavy metals. Especially mercury remains an environmentally problematic metal due to a long-range atmospheric transport potential and long atmospheric residence time of elemental mercury vapors. Total mercury content of mushrooms is species dependent and varies amongst their ecological groups, saprophytic, symbiotic, etc. [53]. Unfortunately, getting any quantitative relationship between metallic elements accumulated in substratum and fruiting bodies is often really difficult [54, 55]. Mineral constituents taken-up by mycelium and acquired in fruiting bodies can be observed by their absolute concentrations values, which are usually expressed on dry weight - due to consensus on 10 % dry matter contents of healthy and of a typical condition specimens. Other point of view on this issue is assessing of accumulate (bio-concentrate) in fruiting bodies elements contained in substratum (e.g. soil, dead plant materials, alive tree, compost), in which mycelium develops [56, 57]. In this study differences in the accumulative abilities of elements in morphological parts of fruiting body were indicated thought estimated cap metal to stipe metal concentration quotients (Q C/S ). Concentrations of elements such as Cd, Cu, Hg, K, Mg, Pb and Zn in caps from both sites were generally greater than those in stipes. For fruiting bodies collected from Augustowska Forest concentration of Al, Cr and Fe were greater in caps than in stipes, while for Co and Na were smaller. Nickel was present in concentrations below method detection limit. Concentrations of the elements (mg kg -1 dry weight) of the Macrolepiota procera s fruiting bodies and soil substratum are given in Table 1. Cadmium, lead and mercury are the most hazardous heavy metal contaminants in foods, which are regulated by law in many countries. These metals were noted in caps in the mean concentrations that varied at two sites between 6.0±3.0 and 16±11 (Cd), 2.7±1.7 and 3.6±1.1 (Pb) and 1.5±0.3 and 4.8±1.1 (Hg) in µg g -1 dry weight, on the average. Parasol Mushroom, like most mushrooms is species-rich in K, Mg and P, which accumulates in the fruiting bodies (Table 1). 3045

3 FIGURE 1 - Location of the sampling sites of Parasol Mushrooms and certain cities (circles) in Poland. Abbreviations: Augustowska Forest in Podlaskie Voivodeship; E in Warmia and Mazury Voivodeship. TABLE 1 - Some metallic elements and phosphorus content of Parasol Mushrooms and soils(µg g -1 dry weight; mean. SD. range and median values) from the Augustowska Forest (; n=16) and region (n=15) and their cap to stipe concentration quotient (Q C/S ) and bioconcentration (BCF) values. Element Site Cap Stipe Q C/S Soil BCF C BCF S Ag 1.3±1.2 ( ) ±2.0 ( ) ±0.33 ( ) ± ( ) ±130 (23-510) ±220 ( ) 390 Al Ba Ca Cd Co 2.1±1.8 ( ) ±210 (42-850) ±65 (58-280) 1.5±1.0 ( ) ±0.5 ( ) ±120 (47-470) ±53 (-260) ±3.0 ( ) ±11 ( ) ±0.32 ( ) ±0.06 ( ) ±0.3 ( ) ±74 (45-320) ±53 (44-460) ±0.7 ( ) ±0.5 ( ) ±230 (120-1) ±320 ( ) ±1.0 ( ) ±0.17 ( ) ±0.39 ( ) ±0.058 ( ) ±0.82 ( ) ±0.7 ( ) ±0.51 ( ) ±0.47 ( ) ±0.36 ( ) ±0.42 ( ) ±0.13 ( ) ±0.7 ( ) ±20 (1.0-63) ±0.33 ( ) ±1.8 ( ) ±50 ( ) ±0.086 ( ) ±0.028 ( ) ±1 (13-15) ±0.07 ( ) ±0.06 ( ) ±5 (74-86) ±1.4 ( ) ±2.8 (1.7-13) ±0.01 ( ) ±33 ( ) 72 17±11 (1.2-39) ±0.02 ( ) ±0.56 ( ) ±0.75 ( )

4 Cr Cu Fe Hg K Mn Mg Na Ni P Pb Rb Sr 0.50±0.17 ( ) ±0.09 ( ) ±58 (89-300) ±32 (69-160) 160±130 (37-570) ±59 (51-260) ±1.1 ( ) ±0.3 ( ) ±6200 ( ) ±18000 ( ) ±14 (18-74) 32 14±6 (9.0-30) ±200 (1300-2) ±190 ( ) 0 240±120 (58-480) ±300 (29-830) ±0.08 ( ) ±0.47 ( ) ±27 (58-160) 120 ±20 (82-140) ±43 (38-210) ±60 (76-250) ±0.5 ( ) ±0.43 ( ) ±4500 ( ) ±6000 ( ) ±44 (14-180) 43 83±40 (43-160) ±190 (430-0) ±94 ( ) ±470 ( ) ±190 ( ) ±0.5 ( ) ±0.37 ( ) ±0.4 ( ) ±0.3 ( ) ±0.7 ( ) ±0.56 ( ) ±0.3 ( ) ±2.0 ( ) ±0.4 ( ) ±0.7 ( ) ±0.62 ( ) ±0.13 ( ) ±0.6 ( ) ±0.5 ( ) ±0.21 ( ) ±2.1 ( ) ±0.0 ( ) ±0.14 ( ) ±0.07 ( ) ±0.05 ( ) ±92 ( ) ±39 (69-210) ±0 ( ) ±0.074 ( ) ±0.024 ( ) ±0.006 ( ) ±48 ( ) 170 ±23 (52-150) 38±3 (35-41) ±140 (590-0) ±130 ( ) ±37 ( ) ±0.04 ( ) ±0.19 ( ) ±5 (96-110) 98 15±2 (13-19) ±1.7 ( ) ±0.4 ( ) ±47 (21-180) ±180 (52-600) 160 < 0.01 < 0.01 ND 0.69±0.02 ( ) 0.69 NA NA < 0.01 < 0.01 ND 10500±1500 ( ) ±0 (3300-6) ±0.5 ( ) ±7 ( ) ±8 (47-80) 59 27±6 (19-34) 28 ND ND ND 2.7±1.7 ( ) ±0.36 ( ) ±2.7 (1.9-13) ±0.6 (8.1-10) ±0.18 ( ) ±0.040 ( ) ±1.1 ( ) ±78 ( ) ±0.6 ( ) ±16 (36-94) ±0.7 ( ) ±0.1 ( ) ±20 (55-130) 76 26±7 (16-37) 25 ND ND ND 0.50±0.36 ( ) ±0.46 ( ) ±0.46 ( ) ±0.03 ( ) ±0.7 ( ) ±0.9 ( ) ±0.15 ( ) ±22 (55-150) 76 Zn ±24 (78-150) 95 Notes: ND (not determined), NA (not analysed) 1.0±0.2 ( ) ±11 (27-65) 29 ±11 (87-120) 0.49±0.20 ( ) ±0.6 ( ) ±0.3 ( ) ±0.3 ( ) ±3 (7.9-22) ±1.6 ( )

5 Data obtained in this study were subjected to statistical analyses including multifunction analysis. In order to demonstrate possible spatial variations in Parasol Mushroom's mineral composition the cluster (CA) and principal components analysis (PCA) have been made. The CA diagram divided all elements in caps and stipes into two main fractions, what did reflect interdependent relationships occurring between them (Fig. 2). The first fraction separated elements such as Cd, K, Mg, Pb and Zn while the second fraction such as Ag, Al, Ba, Ca, Co, Cr, Cu, Fe, Hg, Mn, Na and Sr. In the first fraction a statistically significant positive correlations (p<0.05; the strongest similarity) were found between K and Zn and also Mg and Pb. In the second case, four sub-fractions could be recognized, where relationships between Ag and Na, Al and Fe, Ba and Sr, Ca and Mn, Cu and Hg were the strongest. 120 with each of Mg (PC2, 19.1 % of variation, negative factor coordinate charge) and K (PC3, 10.7 % of variation, positive factor coordinate charge) (Fig. 3). Results showed no statistically significant difference resulting from the sampling site, which means that caps and stipes collected from Augustowska Forest and have the similar content of elements. Two sampling sites differ only in terms of the K content, and the caps and stipes differ only in Mg. Factor 2: 19.10% St3 St3 E St1 E St2 E St C3 C3 E C1 C1 E C2 C2 E 80-3 (Dlink/Dmax)* Factor 1: 57.81% Active FIGURE 3 - Projection of the elements contents in fruiting bodies collected from Augustowska Forest and set on the first and second factor-plane Zn K Pb Mg Cd Hg Cu Co Mn Ca Sr Ba Cr Fe Al Na Ag FIGURE 2 - Cluster analysis tree diagram of the elements concentrations for Parasol Mushroom s fruiting bodies collected from Augustowska Forest (Ward s Method, 1-Pearson r). PCA method was used to graphically illustrate the relationships that exist between elements accumulated in the fruiting bodies and the determination of principal components, which represent the largest part of the variability of the test data set. PCA analysis performed for the specimens (p<0.05) showed the existence of four principal components (PCs), which together explain 93.6 % of all observed variability, while the component PC1 explained 57.8 %, PC %, PC % and PC4 6.2 %. A linear map (projection of the cases on the factor-plane) which was created under the PCA analysis and intended to illustrate which of the elements that determine the PCs are accumulated in similar quantities and whether the sampling site affects their concentrations in the fruiting bodies (Fig. 3). PCA results of the analysis indicate that the greatest differences in concentrations of elements in the fruiting bodies from the two collection sites has been due to closely correlated Al, Ba, Cd, Co, Cr, cu, Fe, Mn, Na and Sr (Fig. 3). Differences in the content of these elements, with negative factor coordinates charge, explain 57.8 % of the variation between fruiting bodies. Fruiting bodies were much less diverse in their content of correlated Factor 2: 23.94% C3 C1 St3 St2 C2 St1 S2 S1 S Factor 1: 63.92% Active FIGURE 4 - Projection of the elements contents in fruiting bodies and soil samples collected from Augustowska Forest set on the first and second factor-plane. In order to observe possible variations in mineral composition of Parasol Mushroom s fruiting bodies and soil substratum in which mycelium develops, also the PCA for samples from Augustowska Forest have been made (Fig. 4). The PCA data revealed that 98.1 % of information regarding the mineral composition variability of Parasol Mushroom s fruiting bodies and soil samples could be described by three principal components (PCs). The elements such as Ag, Al, Ba, Cd, Cu, Fe, Hg, K, Mn, Mg, Na, Ni, P, Rb and Zn caused 63.9 % of the total variability, 3048

6 while the second principal component was determined by Co, Cr, Pb and Sr (23.9 % of total variability). The soils contained at much greater concentrations the elements described by PC1 with positive factor coordinates charge (Al, Ba, Fe, Mn and Ni) and smaller concentrations of elements with negative charge (Ag, Cd, Cu, Hg, K, Mg, Na, p, Rb, Zn) (Fig. 4). The caps and stipes differs less from soil substrate in Co, Cr, Pb and Sr content, amounts of this elements in soil were within mean concentrations determined for all samples (Fig. 4). Many edible mushrooms are known as heavy metals accumulators. It is proved that mushrooms can take-up a large amount of essential and toxic to men trace metals contained in soil, plant or other substratum. The most common way to determine whether the elements of the environment are accumulated by a certain organism is the bioconcentration factor (BCF). In the case of mushrooms will reflect whether a given element is accumulated (BCF >1) or excluded (BCF <1) [16, 51, 58]. The fruiting bodies of Parasol Mushroom tended to accumulate K, Cu, Hg, Na, Rb, Cd, P, Mg, Zn and Ca the BCFs estimated in this study are greater than unity. The highest bioconcentration factor values were for K (BCF 840±140) and Cu (BCF 290±92). Noteworthy is that BCFs for toxic Hg and Cd were 190±48 and 64±33, respectively. The BCFs of Al, Ba, Co, Cr, Fe, Mn and Pb were less than 1, so this species has a tendency to exclude these elements. The European food hygienic standards take into account the content of lead and cadmium in mushrooms, insomuch as to assess possible risk due to intake of some heavy metals accumulated in fruiting bodies, the Reference Dose (RfD) and Provisionally Tolerable Weekly Intake (PTWI) approach have been applied in this study [59-66]. The content of lead in fruiting bodies examined exceed permissible levels set for the European standards (0.3 mg Pb kg -1 fresh weight) for 3 species of cultivated mushrooms. In a similar way, a permissible content of cadmium in 3 species of cultivated mushrooms is 0.2 mg Cd kg -1 fw, while for other mushrooms (excluding these 3 cultivated species, i.e. Agaricus bisporus, Pleurotus ostreatus and Lentinula edodes) is 1.0 mg Cd kg -1 fw [59,60]. Lead contents of caps of Parasol Mushroom in present study, when adjusted to fresh weight data, exceed a tolerance limit for 30% (Augustowska Forest) and 60% (), while of Cd for 70% () specimens. A former value of PTWI for cadmium was 7 µg kg -1 body weight (equivalent to 1 µg kg -1 bw daily) and a revised tolerable weekly intake (PTWI) is 150 µg or 2.5 µg kg -1 bm (equivalent to 0.36 µg kg -1 bm daily) [61, 62]. A single meal usually contains 300 g of mushrooms and a big portion contains 500 g mushrooms (fresh weight) [12]. The median Cd concentrations of 0.62 and 1.6 µg kg -1 fw (assuming 90 % water; Table 1), can be used to estimate Cd intake at the Augustowska Forest () and site (E), respectively. A meal containing 300 or 500 g of fresh caps will result in an intake of 186 and 310 µg Cd () or 480 and 800 µg Cd (), on the average. These values corresponds to 3.1 and 5.2 µg Cd kg -1 bw () and 8.0 and 13 µg kg -1 bw (E) weekly or 0.44 and 0.74 () and 1.1 and 1.9 µg kg -1 bw daily. Hence, a g meal of caps eaten once per week will cause there an intake of Cd at rate from 1.2 to 5.3 -fold exceeding a revised value of PTWI, respectively, if no Cd from other source is taken. Certainly, a frequent eating of a delicious caps of Parasol Mushroom at the sites examined, can lead periodically to an elevated exposure to Cd. The FAO/WHO JECFA a PTWI of Pb is 1.5 mg for a person of 60 kg body weight, which is equivalent to 214 µg daily or 25 µg kg -1 bw weekly or 3.6 µg kg -1 bw daily [63]. A meal made with 300 or 500 g of fresh Parasol Mushroom s caps from the surveyed sites will result in an intake of Pb at the doses of 66 and 110 µg () or 102 and 170 µg (E), on the average. These values corresponds to 1.1 and 1.8 () and 1.7 and 2.8 µg kg -1 bw weekly (E) or 0.16 and 0.26 () and 0.24 and 0.40 µg kg -1 bw (E) daily in a week period. Clearly, in the case of Pb contained in caps, eating of Parasol Mushroom at the region examined is not a controversial issue. For the assessment of possible risks due to intake of Hg accumulated in caps of Parasol Mushroom, a reference dose (RfD; 0.3 µg kg -1 bw daily) and a value of Provisionally Tolerable Weekly Intake (PTWI; 5.0 µg kg -1 bw) have been applied [64-66]. A meal made with 300 or 500 g of fresh Parasol Mushroom s caps from the surveyed sites will result in intake Hg 138 and 230 () or 45 and 75 µg (E), on the average. These values corresponds to 2.3 and 3.8 () 0.75 and 1.2 µg kg -1 bw weekly or 0.33 and 0.55 () and 0.11 and 0.18 µg kg -1 bw daily (E) in a week period, if no Hg from other foods is taken. These estimated doses of Hg intake, when eating once a week caps from a pristine region of Augustowska Forest, are close or above the RfD value of this metal for the non-carcinogenic effects. As was found in a study in Spain [9], the mineral composition of Parasol Mushrooms there and in our study may reflect the species-specific and site related variations, due to occurrence and availability of certain metallic elements present in the soil substrates there. Small concentrations of Cd, Pb, Hg and Ag in the soils in Augustowska Forest sites confirm that area is clean. Great concentration of Fe, Mn, K and Mg may be related to bedrock or original soil material. Dried caps and stipes compared to soil contained greater concentrations of Ag, Cu, K, Hg, Ca, Mg, Zn, Cd and Na, while stipes also P and Rb. In the case of Co, Cr, Ba, Mn and Sr concentrations were similar in the fruiting bodies and upper layer of the soils. 4. CONCLUSION Parasol Mushrooms in unpolluted stands has a high potential to accumulate in fruiting bodies Ag, Hg, Cu and 3049

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9 [60] Commission Regulation (EC) No. 629/2008 of 2 July 2008 amending regulation (EC) No. 1881/2006 setting maximum levels for certain contaminants in foodstuffs. Off J Eur Union L 173, 6 [61] WHO, World Health Organization (2003) Safety evaluation of certain food additives and contaminants. Food Additive Series No. 52. Geneva (Switzerland): Joint FAO/WHO Expert Committee on Food Additives [62] EFSA, European Food Safety Authority (2009) Cadmium in food. EFSA J. 980: contam _op_ej980_cadmium_en_rev.1.pdf?ssbinary=true/ [63] WHO, World Health Organization (1993) Safety evaluation of certain food additives and contaminants, 41 st Report of the Joint FAO/WHO Expert Committee on Food Additives. Technical Report Series No Geneva (Switzerland) [64] JECFA (1978) Evaluation of certain food additives and contaminants. Twenty-second report of the Joint FAO/WHO Expert Committee on Food Additives. WHO Technical Report Series 631 [65] US EPA (1987) Peer review workshop on mercury issues. Environmental criteria and assessment office. Summary report, US Environment Protection Agency, Cincinnati [66] US EPA (2005) Toxicological review of zinc and compounds. In support of summary information on the integrated risk information system. U.S. Environmental Protection Agency Washington DC Received: March 11, 2011 Revised: May 17, 2011 Accepted: May 19, 2011 CORRESPONDING AUTHOR Anna K. Kojta Research Group of Environmental Chemistry, Ecotoxicology & Food Toxicology Institute of Environmental Sciences & Public Health University of Gdańsk 19 Sobieskiego Str Gdańsk POLAND Phone.: , ; Fax: annakojta@gmail.com FEB/ Vol 20/ No 11a/ 2011 pages