Cilj ovog istraživanja je bio da se proceni akumulacija elemenata u četiri komercijalne vrste riba na dva lokaliteta na Dunavu, u Beogradu. U ovom istraživanju, određene su koncentracije 11 elemenata (As, Cd, Cr, Co, Cu, Fe, Hg, Mn, Ni, Se, i  Zn) u tkivima mišića, jetre i škrga sledećih vrsta: mrena (Barbus barbus), deverika (Abramis brama), štuka (Esox lucius) i smuđ (Sander lucioperca). Rezultati su ukazali da je distribucija metala i elemenata u tragovima u različitim tkivima specifična za vrstu. Koncentracije As, Cd, Fe, Hg, i Zn u mišićima riba su bile ispod maksimalno dozvoljenih koncentracija MDK, utvrđenih od strane EU i Republike Srbije. Na osnovu dobijenih rezultata, neophodno je uspostaviti program stalnog monitoringa na Dunavu u Beogradskom regionu

Djikanovic, Vesna

Hegedis, Aleksandar

Jaric, Ivan

Jovicic, Katarina

Lenhardt, Mirjana

Nikolic, Dragica

Skoric, Stefan

Smederevac-Lalic, Marija

Visnjic-Jeftic, Zeljka

English

CaSA NaRA

94 VII INTERNATIONAL CONFERENCE “WATER & FISH” - CONFERENCE PROCEEDINGSAssessMent of tHe MetAL AnD trACe eLeMent Contents in tissUes of foUr CoMMerCiAL fisH sPeCies froM tHe DAnUBe river, BeLGrADeKATARINA JOVIČIĆ1*, ŽELJKA VIŠNJIĆ-JEFTIĆ2, STEFAN SKORIĆ2, MARIJA SMEDEREVAC-LALIĆ2, DRAGICA M. NIKOLIĆ3, VESNA ĐIKANOVIĆ1, IVAN JARIĆ1,4, MIRJANA LENHARDT,1 ALEKSANDAR HEGEDIŠ1,51Institut za Biološka istraživanja“Siniša Stanković”, Univerzitet u Beogradu, Bulevar Despota Stefana 142, 11060 Beograd, Srbija; 2 Institut za Multidisciplinarna istraživanja, Univerzitet u Beogradu, Kneza Višeslava 1, 11030 Beograd, Srbija; 3 Institut za higijenu i tehnologiju mesa, Kaćanskog 13, 11000 Beograd, Srbija; 4leibniz-Institute of freshwater Ecology and Inland fisheries, Müggelseedamm 310, 12587 Berlin, Germany; 5 Biološki fakultet, Univerzitet u Beogradu, Studentski trg 16, 11000 Beograd, SrbijaCorresponding author:e-mail: katarinaj@imsi.rs, katarinaj@imsi.bg.ac.rs phone: PROCENA SADRŽAJA METALA I ELEMENATA U TKIVIMA 4 KOMERCIJALNE VRSTE RIBA IZ DUNAVA KOD BEOGRADAApstraktCilj ovog istraživanja je bio da se proceni akumulacija elemenata u četiri komercijalne vrste riba na dva lokaliteta na Dunavu, u Beogradu. U ovom istraživanju, određene su kon-centracije 11 elemenata (As, Cd, Cr, Co, Cu, Fe, Hg, Mn, Ni, Se, i  Zn) u tkivima mišića, jetre i škrga sledećih vrsta: mrena (Barbus barbus), deverika (Abramis brama), štuka (Esox lucius) i smuđ (Sander lucioperca). Rezultati su ukazali da je distribucija metala i eleme-nata u tragovima u različitim tkivima specifična za vrstu. Koncentracije As, Cd, Fe, Hg, i Zn u mišićima riba su bile ispod maksimalno dozvoljenih koncentracija MDK, utvrđenih od strane EU i Republike Srbije. Na osnovu dobijenih rezultata, neophodno je uspostaviti program stalnog monitoringa na Dunavu u Beogradskom regionu.Ključne reči: Dunav, riba, metal, teški metali, ICP-MSKeywords: Danube River, fish, metal, trace element, ICP-MS.VII INTERNATIONAL CONFERENCE “WATER & FISH” - ZBORNIK PREDAVANJA 95INTRODUCTIONThe Danube River is polluted from various sources, such as industry, agriculture and energetics (Milanović et al. 2010). Metals and trace elements are considered to be among major pollutants in the Danube Basin (Teodorović 2009). Fish, due to their position at the top of the aquatic food chains, are able to accumulate metals and trace elements at such levels that could pose a threat not only to the fish but also to the human population (Yilmaz et al.2007). Therefore, fish are considered as one of the best pollution indicators of aquatic ecosystems. Accumulation levels and distribution patterns of metals and trace elements differ depending on fish species as well as on fish tissue (Višnjić-Jeftić et al. 2010, Jarić et al. 2011). Previous studies of metal and trace element concentrations (Jovičić et al. 2014; Lenhardt et al. 2012) indicated great importance of fish tissue contamination monitoring, given that toxic chemicals can produce adverse effects on consumers.In the present study, 11 elements were analyzed in the muscle, liver and gills of the four fish species from the Danube River. The main aim of the present study was to assess poten-tial impacts of accumulation patterns in fish tissues in the Danube River.MATERIAL AND METHODSSample collectionFish specimens were collected from the two localities on the Danube River on the teri-tory of the city of Belgrade, Serbia (downstream Danube section 44° 52’ 29’’ N, 20° 22’01’’ E; upstream Danube section 44° 49’ 57’’ N, 20° 27’ 47’’E). Samples were collected between November 2012 and September 2013. Samples of the muscle, gills and liver of the following fish species were collected: barbel (Barbus barbus), freshwater bream (Abramis brama), northern pike (Esox lucius) and pike-perch (Sander lucioperca). Sample preparation and analysisSamples were dried by Freeze Dryers Rotational-Vacuum-Concentrator, GAMMA 1-16 LSC, Germany, and sample portions of approximately 0.3 g (dry weight) were afterwards processed in a microwave digester (speedwave™ MWS-3+; Bergof Products + Instruments GmbH, Eningem, Germany), using 6 ml of 65% HNO3 and 4 ml of 30% H2O2 (Merck su-prapure) at a food temperature program (100 – 170°C). Potential presence of trace elements in chemicals used in digestion was resolved by using a number of blank samples. Following a cooling to room temperature, the digested samples were diluted with distilled water to a total volume of 25 ml. The analysis was performed by inductively-coupled plasma mass spectrometry (ICP-MS). Instrument was Thermo Scientific (Bremen, Germany), model iCAP Q. The following iso-topes were measured: 52Cr, 55Mn, 57Fe, 59Co, 60Ni, 63Cu, 66Zn, 75As, 77Se, and 111Cd. The quality of the analytical process was controlled by the analysis of reference material in each sample batch (NIST CRM 1577c). All concentrations were expressed as µg g-1 dry weight (dw).Mercury (Hg) was measured using cold vapor technique by atomic absorption spec-trometer Varian “SpectrAA 220” with VGA 77 hydride system. The wavelenght line for Hg was 257.3 nm. The quality of the analytical process was controlled by the analysis of reference material in each sample batch (BCR-186R). 96 VII INTERNATIONAL CONFERENCE “WATER & FISH” - CONFERENCE PROCEEDINGSConcentrations in fish meat (i.e., muscle samples) were compared with the maximum al-lowed concentrations (MAC) for the utilization in human diet, established by the European Union (EU) and the national legislation (EuropEan Commission rEgulation, 2006, offiCial gazEttE of rs 2011). Normality of distribution of analyzed data samples was tested by Kolmogorov-Smir-nov test. Assessment of the differences among the groups was performed by means of the Kruskal-Wallis H test, which was followed by comparisons of particular pairs of samples by Mann-Whitney U test (p<0.05 was used as a threshold value).RESULTSMetal and trace element concentrations in analyzed tissues of the four fish species on the two studied sites are presented in Table 1 and Table 2. There were only a few differences between elemental concentrations among the two studied localities. In the muscle of the barbel, As and Ni concentrations were higher in the upstream Dan-ube section, while Se concentrations were higher in the downstream section. Concentra-tions of these elements in the liver were higher in the downstream Danube section. Gills had a higher As concentrations in downstream section, while Ni and Se concentrations were lower (Table 1 and 2). table 1. Heavy metal and trace element concentrations in muscle, liver and gills of the four fish species at the upstream Danube section (means ± standard deviation). Concentrations are expressed as μg g-1 dry weight, nd indicates the values below the detect on threshold.Tissue element BarbelFreshwater Breamnorthern Pike Pike-PerchMuscle As 0.289±0.151 0.325±0.145 0.263±0.090 0.548±0.282Cd nd nd 0.010±0.011 0.010±0.367Cr 0.165±0.056 0.183±0.103 0.416±0.786 0.152±0.079Co nd 0.030±0.023 nd 0.010±0.022Cu 1.579±0.365 1.973±1.055 2.477±1.054 1.237±0.439Fe 16.193±8.552 25.784±25.788 23.307±13.914 59.059±91.085Hg 0.962±0.359 0.447±0.391 1.351±0.644 1.790±0.367Mn 1.131±0.479 1.665±0.845 1.116±0.604 4.174±7.435Ni 0.410±0.371 0.220±0.145 0.340±0.315 0.210±0.234Se 1.118±0.624 1.884±0.881 1.089±0.237 1.604±0.907Zn 19.430±5.503 36.675±9.827 48.429±22.487 47.312±19.871Liver As 3.043±0.397 7.522±4.008 2.338±1.526 7.617±5.319Cd 0.060±0.031 0.180±0.216 0.040±0.050 0.340±0.470Cr 0.111±0.061 0.094±0.106 0.175±0.048 0.230±0.255Co 0.050±0.019 0.170±0.063 0.060±0.010 0.430±0.334Cu 22.069±8.607 69.331±51.031 27.451±10.203 17.066±7.012VII INTERNATIONAL CONFERENCE “WATER & FISH” - ZBORNIK PREDAVANJA 97Fe 139.626±26.007 258.193±165.011 126.522±64.167 425.626±421.694Hg 0.246±0.119 0.156±0.149 0.350±0.358 0.901±0.435Mn 2.052±0.473 16.195±26.174 11.431±13.850 6.473±4.671Ni 0.350±0.611 0.460±0.698 0.530±0.168 0.520±0.565Se 0.325±0.105 0.310±0.141 0.491±0.298 3.442±1.591Zn 72.689±19.463 133.630±70.709 151.546±31.563 120.439±74.104Gills As 2.566±0.746 3.197±0.933 1.786±0.855 1.665±0.420Cd 0.030±0.011 0.020±0.002 0.020±0.007 0.030±0.024Cr 0.326±0.088 0.332±0.122 0.267±0.068 0.385±0.343Co 0.060±0.010 0.220±0.080 0.040±0.011 0.050±0.041Cu 2.590±0.161 2.550±0.668 2.053±0.293 1.708±0.654Fe 279.985±74.612 353.657±109.036 194.362±49.913 136.979±102.696Hg 0.161±0.048 0.070±0.045 0.143±0.137 0.413±0.245Mn 12.627±7.102 37.221±12.816 28.383±13.795 11.372±7.463Ni 0.370±0.244 0.820±0.768 0.520±0.303 0.580±0.474Se 0.253±0.067 0.267±0.095 0.179±0.075 0.936±0.640Zn 73.368±10.987 107.995±36.395 494.037±79.150 76.629±36.325Higher Ni concentrations in the muscle of the freshwater bream were observed in the downstream section, while concentrations of Se were higher in the upstream section. Con-centrations of As and Mn in the liver were higher in the downstream section, while concen-trations of the Ni and Se were lower at this locality. Gills had higher As and Mn concen-trations in the downstream section, while Cr, Ni and Se concentrations were higher in the upstream Danube section (Table 1 and 2). Northern pike muscle had higher As concentrations at the downstream section.  Liver had higher As, Cu and Mn concentrations and lower Se concentrations were also observed in the downstream section. Concentrations of As and Mn in gills were higher in specimens from downstream section, while Se concentrations were lower (Table 1 and 2).table 2. Heavy metal and trace element concentrations in muscle, liver and gills of the four fish species at the downstream Danube section (means ± standard deviation). Con-centrations are expressed as μg g-1 dry weight, nd indicates the values below the detection threshold.tissue element BarbelFreshwater Breamnorthern Pike Pike-PerchMuscle As 0.289±0.151 0.325±0.145 0.263±0.090 0.548±0.282Cd nd nd 0.010±0.011 0.010±0.367Cr 0.165±0.056 0.183±0.103 0.416±0.786 0.152±0.079Co nd 0.030±0.023 nd 0.010±0.022Cu 1.579±0.365 1.973±1.055 2.477±1.054 1.237±0.43998 VII INTERNATIONAL CONFERENCE “WATER & FISH” - CONFERENCE PROCEEDINGSFe 16.193±8.552 25.784±25.788 23.307±13.914 59.059±91.085Hg 0.962±0.359 0.447±0.391 1.351±0.644 1.790±0.367Mn 1.131±0.479 1.665±0.845 1.116±0.604 4.174±7.435Ni 0.410±0.371 0.220±0.145 0.340±0.315 0.210±0.234Se 1.118±0.624 1.884±0.881 1.089±0.237 1.604±0.907Zn 19.430±5.503 36.675±9.827 48.429±22.487 47.312±19.871Liver As 3.043±0.397 7.522±4.008 2.338±1.526 7.617±5.319Cd 0.060±0.031 0.180±0.216 0.040±0.050 0.340±0.470Cr 0.111±0.061 0.094±0.106 0.175±0.048 0.230±0.255Co 0.050±0.019 0.170±0.063 0.060±0.010 0.430±0.334Cu 22.069±8.607 69.331±51.031 27.451±10.203 17.066±7.012Fe 139.626±26.007 258.193±165.011 126.522±64.167 425.626±421.694Hg 0.246±0.119 0.156±0.149 0.350±0.358 0.901±0.435Mn 2.052±0.473 16.195±26.174 11.431±13.850 6.473±4.671Ni 0.350±0.611 0.460±0.698 0.530±0.168 0.520±0.565Se 0.325±0.105 0.310±0.141 0.491±0.298 3.442±1.591Zn 72.689±19.463 133.630±70.709 151.546±31.563 120.439±74.104Gills As 2.566±0.746 3.197±0.933 1.786±0.855 1.665±0.420Cd 0.030±0.011 0.020±0.002 0.020±0.007 0.030±0.024Cr 0.326±0.088 0.332±0.122 0.267±0.068 0.385±0.343Co 0.060±0.010 0.220±0.080 0.040±0.011 0.050±0.041Cu 2.590±0.161 2.550±0.668 2.053±0.293 1.708±0.654Fe 279.985±74.612 353.657±109.036 194.362±49.913 136.979±102.696Hg 0.161±0.048 0.070±0.045 0.143±0.137 0.413±0.245Mn 12.627±7.102 37.221±12.816 28.383±13.795 11.372±7.463Ni 0.370±0.244 0.820±0.768 0.520±0.303 0.580±0.474Se 0.253±0.067 0.267±0.095 0.179±0.075 0.936±0.640Zn 73.368±10.987 107.995±36.395 494.037±79.150 76.629±36.325Muscle of the pike-perch had Fe, Mn and Se concentrations higher in specimens from the downstream section, while As and Ni concentrations were lower. Concentrations of As and Cu were higher in the liver tissue from the downstream section specimens, while Ni concentrations were higher in specimens from the upstream section. In gills, concentrati-ons of As were higher in specimens inhabiting the downstream Danube, but they had lower Cr concentrations (Table 1 and 2). Concentrations of As, Cd, Fe, Hg, and Zn in the fish muscle were below the MAC established by both the EU and the Republic of Serbia.VII INTERNATIONAL CONFERENCE “WATER & FISH” - ZBORNIK PREDAVANJA 99DISCUSSIONDistribution of metal and trace elements in different tissues showed species-specific patterns. In the present study, the species with the highest overall accumulation in the mus-cle tissue in the downstream section was the pike-perch. Barbel had the highest concentra-tions of the metal and trace elements in the muscle at the upstream section. However, con-centrations of the As, Cd, Cu, Fe and Zn were below MAC in all analyzed muscle samples, which indicates that the meat of the studied fish species can be utilized in the human diet. The gills, due to their direct contact with the water, are considered to be the main site of the metal uptake and can indicate the water as a main source of the contamination (Storelli et al. 2007). In the current study, gills had the highest Cr, Zn and Fe concentrations. Previous research (Lenhardt et al. 2012) in the Belgrade Danube section, showed some differences from our findings. Since freshwater bream was analyzed in both studies, we observed that overall metal and trace element concentration in liver and muscle was higher in the current study, which can be potentially explained by recent pollution emissions. Ad-ditionally, there were no significant differences in the overall gill metal and trace element concentrations.Since liver is considered as a tissue with the highest accumulation ability (Uysal et al. 2009) and our study showed increase in metal and trace element concentrations, we strong-ly emphasize the necessity to establish continuous monitoring activities in the future, due to possible environmental deterioration. It is important to establish permanent monitoring of metals and trace elements in these localities, as well as to establish a system of suitable indicator species. ACKNOWLEDGEMENTSThis study was supported by the Ministry of Education, Science, and Technological Development of the Republic of Serbia, Project No. TR37009 and Project No. 173045.REFERENCESEuropean Commission Regulation, Setting maximum levels for certain contaminants in foodstuffs, Official Journal of the European Union No 1881/2006, 19 December 2006, 2006.Jarić, I., Višnjić-Jeftić, Ž., Cvijanović, G., Gačić, Z., Jovanović, Lj., Skorić, S., Lenhardt, M. (2011): Determination of differential heavy metal and trace element accumulation in liver, gills, intestine and muscle of sterlet (Acipenser ruthenus) from the Danube river in Serbia by ICP-OES, Microchem. J. 98:77-81.Jovičić, K., Nikolić, M.D., Višnjić-Jeftić, Ž., Đikanović, V., Skorić, S., Stefanović, M.S., Lenhardt, M., Hegediš., A, Krpo-Ćetković, J., Jarić, I. (2014): Mapping differential ele-mental accumulation in fish tissues: assessment of metal and trace element concentrations in wels catfish (Silurus glanis) from the Danube River by ICP-MS, Environ Sci Pollut Res 1-8.Lenhardt, M., Jarić, I., Višnjić-Jeftić, Ž., Skorić, S.,.Gačić, Z,, Pucar, M., Hegediš, A. (2012): Concentrations of 17 elements in muscle, gills, liver and gonads of five economi-cally important fish species from the Danube River, Knowl. Manag. Aquat. Ecosyst. 407 (2) 1-10.100 VII INTERNATIONAL CONFERENCE “WATER & FISH” - CONFERENCE PROCEEDINGSMilanović, A., Kovačević-Majkić, J., Milivojević, M. (2010): Water quality analysis of Danube River in Serbia-pollution and protection problems, Bulletin of the Serbian Geo-graphical Society 90:59-68.Official Gazzette of RS, Regulation on quantity of pesticides, metals, metalloids, and other toxic substances, chemotherapeutics, anabolics, and other substances which can be found in food, Official Gazzette of RS No 28/2011, 2011.Storelli, M.M., Barone, G., Storelli, A., Marcotrigiano, G.O. (2006): Trace metals in tissues of Mugilids (Mugil auratus, Mugil capito, and Mugil labrosus) from the Mediterra-nean Sea, Bull Environ Contam Tox  77:43-50.Teodorović, I. (2009): Ecotoxicological research and related legislation in Serbia, Envi-ron. Sci. Pollut. Res. 16 (Suppl. 1) S123-S129.Uysal, K., Köse, E., Bülbül, M., Dönmez, M., Erdoğan, Y., Koyun, M., Ömeroğlu, Ç., Özmal, F. (2009): The comparison of heavy metal accumulation ratios of some fish species in Enne Dame Lake (Kütahya/Turkey), Environ Monit Assess 157:355-362.Višnjić-Jeftić, Ž., Jarić, I., Jovanović, Lj., Skorić, S., Smederevac-Lalić, M., Nikčević, M., Lenhardt, M. (2010): Heavy metal and trace element accumulation in muscle, liver and gills of the Pontic shad (Alosa immaculata Bennet 1835) from the Danube River (Serbia), Microchem J. 95:341-344.Yilmaz, F., Özdemir, N., Demirak, A., Tuna, A.L. (2007): Heavy metal levels in two fish species leuscius cephalus and lepomis gibbosus, Food Chem. 100 830-835.

Procena sadržaja metala i elemenata u tkivima 4 komercijalne vrste riba iz Dunava kod Beograda

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Procena sadržaja metala i elemenata u tkivima 4 komercijalne vrste riba iz Dunava kod Beograda

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