Environmental Science and Pollution Research

, Volume 24, Issue 30, pp 23977–23991 | Cite as

Apex predatory mammals as bioindicator species in environmental monitoring of elements in Dinaric Alps (Croatia)

  • Maja Lazarus
  • Ankica Sekovanić
  • Tatjana Orct
  • Slaven Reljić
  • Josip Kusak
  • Jasna Jurasović
  • Đuro Huber
Research Article


Tissue element investigations of apex terrestrial mammals are very scarce in Europe. We quantified 16 essential and nonessential elements in the kidney cortex, liver, and muscle tissue of 467 brown bears (Ursus arctos), 125 gray wolves (Canis lupus), one Eurasian lynx (Lynx lynx), and three golden jackals (Canis aureus) from Croatia by inductively coupled plasma mass spectrometry (ICP-MS). Renal cadmium (0.6% of animals) and lead (1%) and hepatic lead (5%) were found in toxicologically relevant levels for mammals only in bears, while the other elements were within normal range. The association of age, sex, season, and region with measured tissue elements in bear and wolf was estimated by multiple regression analyses. Age-related accumulation of cadmium was observed in bears and wolves. Lead tissue content increased with the age of bears but declined in wolves. Female bears and wolves had higher arsenic, iron, and thallium than males in some tissues. Also, cadmium, mercury, copper, zinc, selenium, molybdenum, and uranium were more abundant only in female bears. Male bears had higher potassium, zinc, and magnesium, while male wolves had higher calcium in some tissues compared to female wolves. Seasonal differences were mainly observed for bears’ tissues and region-specific differences only in wolves. The bear kidneys had the highest levels of cobalt, copper, molybdenum, cadmium, and lead among the four studied species. The element levels reported for bears and wolves represent baseline values for the Dinaric population.


Bear Wolf Lynx Jackal Trace elements Croatia 



The help of local hunters and experts in the collection of samples is gratefully acknowledged. The authors wish to thank Dr Z. Kljaković-Gašpić and anonymous reviewers for valuable comments on the manuscript and Mr Makso Herman for language editing.

Funding information

This study was supported by the Ministry of Science, Education and Sports of the Republic of Croatia (grant no. 022-0222148-2135) and by the European Commission under the “HUNT” project of the 7th Framework Programme for Research and Technological Development (grant no. 212160). Neither the European Commission nor any person acting on behalf of the Commission is responsible for the use made of the information. The views expressed in this publication are the sole responsibility of the authors and do not necessarily reflect the views of the European Commission. The Veterinary Faculty team was additionally supported by the Research Council of Norway under the project “The role of natural resources in sustainable rural livelihoods in the western Balkans. The distribution and flow of costs and benefits” (application no. ES459363).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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  1. AMAP (Arctic Monitoring and Assessment Programme) (2005) AMAP assessment 2002: heavy metals in the Arctic. AMAP, Oslo Accessed 10 Apr 2017Google Scholar
  2. Barja I (2009) Prey and prey-age preference by the Iberian wolf Canis lupus signatus in a multiple-prey ecosystem. Wildl Biol 15(2):147–154CrossRefGoogle Scholar
  3. Berne Convention, Convention on the Conservation of European Wildlife and Natural Heritage (1979) Council of Europe, Bern, Switzerland. Accessed 10 Apr 2017
  4. Bilandžić N, Sedak M, Dokić M, Šimić B (2010) Wild boar tissue levels of cadmium, lead and mercury in seven regions of continental Croatia. Bull Environ Contam Toxicol 84:738–743CrossRefGoogle Scholar
  5. Bilandžić N, Dežđek D, Sedak M, Dokić M, Šimić B, Rudan N, Brstilo M, Lisicin T (2012) Trace elements in tissues of wild carnivores and omnivores in Croatia. Bull Environ Contam Toxicol 88(1):94–99CrossRefGoogle Scholar
  6. Burger J (2007) A framework and methods for incorporating gender-related issues in wildlife risk assessment: gender-related differences in metal levels and other contaminants as a case study. Environ Res 104:153–162CrossRefGoogle Scholar
  7. Burger J, Fossi C, McClellan-Green P, Orlando EF (2007) Methodologies, bioindicators, and biomarkers for assessing gender-related differences in wildlife exposed to environmental chemicals. Environ Res 104(1):135–152CrossRefGoogle Scholar
  8. CBS (Croatian Bureau of Statistics) (2005) Statistical yearbook of the Republic of Croatia. Zagreb, p 709. Accessed 20 Jan 2017
  9. Čelechovská O, Literák I, Ondruš S, Pospíšil Z (2006) Heavy metals in brown bears from the Central European Carpathians. Acta Vet Brno 75:501–506CrossRefGoogle Scholar
  10. Census of Population, Households and Dwellings 2011 (2012) Croatian Bureau of Statistics, Zagreb Accessed 20 Jan 2017
  11. Chapron G et al (2014) Recovery of large carnivores in Europe’s modern human-dominated landscapes. Science 346:1517–1519CrossRefGoogle Scholar
  12. Cicnjak L, Huber Đ, Roth HU, Ruff RL, Vinovrški Z (1987) Food habits of brown bears in Plitvice Lakes National Park, Yugoslavia. Int Conf Bear Res Manage 7:221–226Google Scholar
  13. Ćirović D, Gizejewska A, Jovanović V, Penezić A, Milenković M, Vujošević M, Blagojević J (2015) Concentration of selected trace elements in the golden jackal (Canis aureus L., 1758) population from Serbia. Acta Zool Bulg 67(3):409–414Google Scholar
  14. Ćirović D, Penezić A, Krofel M (2016) Jackals as cleaners: ecosystem services provided by a mesocarnivore in human-dominated landscapes. Biol Conserv 199:51–55CrossRefGoogle Scholar
  15. Cooke JA, Johnson MS (1996) Cadmium in small mammals. In: Beyer WN, Heinz GH, Redmon-Norwood AW (eds) Environmental contaminants in wildlife: interpreting tissue residues. CRC, Boca Raton, pp 377–405Google Scholar
  16. Council Directive 92/43/EEC of 21 May 1992 on the conservation of natural habitats and of wild fauna and flora. Available from:
  17. Crête M, Nault R, Walsh P, Benedetti JL, Lefebvre MA, Weber JP, Gagnon J (1989) Variation in cadmium content of caribou tissues from northern Québec. Sci Total Environ 80(2–3):103–112CrossRefGoogle Scholar
  18. Croatian Water (2014) Influence of agriculture on surface and underground water contamination in the Republic of Croatia. Croatian, p 29Google Scholar
  19. Danielsson R, Frank A (2009) Cadmium in moose kidney and liver—age and gender dependency, and standardisation for environmental monitoring. Environ Monit Assess 157(1–4):73–88CrossRefGoogle Scholar
  20. Elinder CG, Nordberg M, Palm B, Piscator M (1981) Cadmium, zinc, and copper in horse liver and in horse liver metallothionein: comparisons with kidney cortex. Environ Res 26(1):22–32CrossRefGoogle Scholar
  21. Falandysz J (1994) Some toxic and trace metals in big game hunted in the northern part of Poland in 1987–1991. Sci Total Environ 141:59–73CrossRefGoogle Scholar
  22. Farkas A, Bidló A, Bolodár-Varga B, Jánoska F (2017) Accumulation of metals in liver tissues of sympatric golden jackal (Canis aureus) and red fox (Vulpes vulpes) in the southern part of Romania. Bull Environ Contam Toxicol 98(4):513–520CrossRefGoogle Scholar
  23. Flaten TP, Andersen LH, Holsen AMH, Ottemo VG, Pedersen HC, Steinnes E, Lierhagen S, Jenssen BM (2008) A nationwide survey of trace elements in lynx, wolverines, wolves, and brown bears in Norway. Cell Biol Toxicol 24(Suppl 1):6–7Google Scholar
  24. Gamberg M, Braune BM (1999) Contaminant residue levels in arctic wolves (Canis lupus) from the Yukon Territory, Canada. Sci Total Environ 243-244:329–338CrossRefGoogle Scholar
  25. Gamberg M, Palmer M, Roach P (2005) Temporal and geographic trends in trace element concentrations in moose from Yukon, Canada. Sci Total Environ 351-352:530–538CrossRefGoogle Scholar
  26. Gamberg M, Chételat J, Poulain AJ, Zdanowicz C, Zheng J (2015) Mercury in the Canadian Arctic terrestrial environment: an update. Sci Total Environ 509–510:28–40CrossRefGoogle Scholar
  27. García MHM, Moreno DH, Rodríguez FS, Beceiro AL, Álvarez LEF, López MP (2010) Sex- and age-dependent accumulation of heavy metals (Cd, Pb and Zn) in liver, kidney and muscle of roe deer (Capreolus capreolus) from NW Spain. J Environ Sci Health Part A 46:109–116CrossRefGoogle Scholar
  28. Gipson PS, Ballard WB, Nowak RM, Mech LD (2000) Accuracy and precision of estimating age of gray wolves by tooth wear. J Wildl Manag 46:752–758CrossRefGoogle Scholar
  29. Gnamuš A, Byrne AR, Horvat M (2000) Mercury in the soil-plant-deer-predator food chain of a temperate forest in Slovenia. Environ Sci Technol 34:3337–3345CrossRefGoogle Scholar
  30. Golubkina NA, Kovalsky YG, Senkevich OA (2011) Selenium content in muscle tissue of wildlife in Khabarovsk Territory. Russ Agric Sci 37(5):419–422CrossRefGoogle Scholar
  31. Goyer RA (1997) Toxic and essential metal interactions. Annu Rev Nutr 17:37–50CrossRefGoogle Scholar
  32. Halamić J, Miko S (2009) Geochemical atlas of the Republic of Croatia. Croatian Geological Survey, ZagrebGoogle Scholar
  33. Hassan AA, Brustad M, Sandanger TM (2012) Concentrations and geographical variations of selected toxic elements in meat from semi-domesticated reindeer (Rangifer tarandus tarandus L.) in mid- and northern Norway: evaluation of risk assessment. Int J Environ Res Public Health 9(5):1699–1714CrossRefGoogle Scholar
  34. Herceg Romanić S, Klinčić D, Kljaković-Gašpić Z, Kusak J, Reljić S, Huber Đ (2015) Organochlorine pesticides and polychlorinated biphenyl congeners in wild terrestrial mammals from Croatia: interspecies comparison of residue levels and compositions. Chemosphere 137:52–58CrossRefGoogle Scholar
  35. Hermoso de Mendoza García M, Hernández Moreno D, Soler Rodríguez F, López Beceiro A, Fidalgo Alvarez LE, Pérez López M (2011) Sex- and age-dependent accumulation of heavy metals (Cd, Pb and Zn) in liver, kidney and muscle of roe deer (Capreolus capreolus) from NW Spain. J Environ Sci Health A Tox Hazard Subst Environ Eng 46(2):109–116CrossRefGoogle Scholar
  36. Hernández-Moreno D, de la Casa RI, Fidalgo LE, Llaneza L, Soler Rodríguez F, Pérez-López M, López-Beceiro A (2013) Noninvasive heavy metal pollution assessment by means of Iberian wolf (Canis lupus signatus) hair from Galicia (NW Spain): a comparison with invasive samples. Environ Monit Assess 185(12):10421–10430CrossRefGoogle Scholar
  37. Hoffmann SR, Blunck SA, Petersen KN, Jones EM, Koval JC, Misek R, Frick JA, Cluff HD, Sime CA, McNay M, Beckman KB, Atkinson MW, Drew M, Collinge MD, Bangs EE, Harper RG (2010) Cadmium, copper, iron, and zinc concentrations in kidneys of grey wolves, Canis lupus, from Alaska, Idaho, Montana (USA) and the Northwest Territories (Canada). Bull Environ Contam Toxicol 85(5):481–485CrossRefGoogle Scholar
  38. Huber Đ, Roth HU (1997) Denning of brown bears in Croatia. Int Conf Bear Res Manage 9(2):79–83Google Scholar
  39. Kaczensky P, Chapron G, Arx M, Huber Đ, Andrén H, Linnell J (2012) Status, management and distribution of large carnivores—bear, lynx, wolf & wolverine—in Europe. Istituto di Ecologia Applicata and the IUCN/SSC Large Carnivore Initiative for Europe report prepared for the European Commission (contract N°070307/2012/629085/SER/B3), Rome, Italy, 72 ppGoogle Scholar
  40. Kramárová M, Massányi P, Jančová A, Toman R, Slamečka J, Tataruch F, Kováčik J, Gašparík J, Nad P, Skalická M, Koréneková B, Jurčík R, Čuboň J, Haščík P (2005) Concentration of cadmium in the liver and kidneys of some wild and farm animals. Bull Vet Inst Pulawy 49:465–469Google Scholar
  41. Krofel M, Huber Đ, Kos I (2011) Diet of Eurasian lynx Lynx lynx in Northern Dinaric Mountains (Slovenia and Croatia): importance of edible dormouse Glis glis as alternative prey. Acta Theriol 56(4):315–322CrossRefGoogle Scholar
  42. Kusak J (2002) Conditions for life of wolves (Canis lupus L.) in Croatia. Dissertation, Faculty of Science, University of ZagrebGoogle Scholar
  43. Kusak J, Huber Đ (1998) Brown bear habitat quality in Gorski Kotar, Croatia. Ursus 10:281–291Google Scholar
  44. Larison JR, Likens GE, Fitzpatrick JW, Crock JG (2000) Cadmium toxicity among wildlife in the Colorado Rocky Mountains. Nature 406(6792):181–183CrossRefGoogle Scholar
  45. Larter NC, Macdonald CR, Elkin BT, Wang X, Harms NJ, Gamberg M, Muir DC (2016) Cadmium and other elements in tissues from four ungulate species from the Mackenzie Mountain region of the Northwest Territories, Canada. Ecotoxicol Environ Saf 132:9–17CrossRefGoogle Scholar
  46. Lazarus M, Orct T, Blanuša M, Vicković I, Šoštarić B (2008) Toxic and essential metal concentrations in four tissues of red deer (Cervus elaphus) from Baranja, Croatia. Food Addit Contam Part A 25(3):270–283CrossRefGoogle Scholar
  47. Lazarus M, Prevendar Crnić A, Bilandžić N, Kusak J, Reljić S (2014a) Cadmium, lead, and mercury exposure assessment among Croatian consumers of free-living game. Arch Ind Hyg Toxicol 65:281–292Google Scholar
  48. Lazarus M, Sekovanić A, Reljić S, Kusak J, Kovačić J, Orct T, Jurasović J, Huber Đ (2014b) Selenium in brown bears (Ursus arctos) from Croatia: relation to cadmium and mercury. J Environ Sci Health Part A 49:1392–1401CrossRefGoogle Scholar
  49. Lockhart JM, Siddiqui S, Loughry WJ, Bielmyer-Fraser GK (2016) Metal accumulation in wild-caught opossum. Environ Monit Assess 188(6):317. CrossRefGoogle Scholar
  50. Ma WC (1996) Lead in mammals. In: Beyer WN, Heinz GH, Redmon-Norwood AW (eds) Environmental contaminants in wildlife: interpreting tissue residues. CRC, Boca Raton, pp 281–296Google Scholar
  51. Markov G, Kocheva M, Gospodinova M (2016) Assessment of heavy metal accumulation in the golden jackal (Canis aureus) as a possible bioindicator in an agricultural environment in Bulgaria. Bull Environ Contam Toxicol 96(4):458–464CrossRefGoogle Scholar
  52. Medvedev N (1999) Levels of heavy metals in Karelian wildlife, 1989–91. Environ Monit Assess 56:177–193CrossRefGoogle Scholar
  53. Millán J, Mateo R, Taggart MA, López-Bao JV, Viota M, Monsalve L, Camarero PR, Blázquez E, Jiménez B (2008) Levels of heavy metals and metalloids in critically endangered Iberian lynx and other wild carnivores from southern Spain. Sci Total Environ 399(1–3):193–201CrossRefGoogle Scholar
  54. Neila C, Hernández-Moreno D, Fidalgo LE, López-Beceiro A, Soler F, Pérez-López M (2017) Does gender influence the levels of heavy metals in liver of wild boar? Ecotoxicol Environ Saf 140:24–29CrossRefGoogle Scholar
  55. Nordberg M (1998) Metallothioneins: historical review and state of knowledge. Talanta 46:243–254CrossRefGoogle Scholar
  56. OG (Official Gazette) (2005) Hunting Act. Official Gazette No 140/05, 2641 (in Croatian). Accessed 16 Jan 2017
  57. OG (Official Gazette) (2013) Regulation on the strictly protected species. Official Gazette No 144/13, 3086 (in Croatian). Accessed 16 Jan 2017
  58. Pokorny B, Ribaric-Lasnik C (2002) Seasonal variability of mercury and heavy metals in roe deer (Capreolus capreolus) kidney. Environ Pollut 117(1):35–46CrossRefGoogle Scholar
  59. Prunescu C, Serban-Parau N, Broc JH, Vaughan DM, Prunescu P (2003) Liver and kidney structure and iron content in Romanian brown bears (Ursus arctos) before and after hibernation. Comp Biochem Physiol A Mol Integr Physiol 134(1):21–26CrossRefGoogle Scholar
  60. Puls R (1994) Mineral levels in animal health. Diagnostic data. Sherpa International, ClearbrookGoogle Scholar
  61. Radović A, Kovačić D (2010) Diet composition of the golden jackal (Canis aureus L.) on the Pelješac Peninsula, Dalmatia, Croatia. Period Biol 112:219–224Google Scholar
  62. Romero MB, Polizzi P, Chiodi L, Das K, Gerpe M (2016) The role of metallothioneins, selenium and transfer to offspring in mercury detoxification in Franciscana dolphins (Pontoporia blainvillei). Mar Pollut Bull 109(1):650–654CrossRefGoogle Scholar
  63. Sato I, Yamauchi K, Tsuda S (2016) Long-term survey of cadmium and lead contamination in Japanese black bears captured in Iwate Prefecture, Japan. Bull Environ Contam Toxicol 97(6):806–812CrossRefGoogle Scholar
  64. Scheuhammer AM (1991) Effects of acidification on the availability of toxic metals and calcium to wild birds and mammals. Environ Pollut 71(2–4):329–375CrossRefGoogle Scholar
  65. Shore RF, Casulli A, Bologov V, Wienburg CL, Afsar A, Toyne P, Dell'Omo G (2001) Organochlorine pesticide, polychlorinated biphenyl and heavy metal concentrations in wolves (Canis lupus L. 1758) from north-west Russia. Sci Total Environ 280(1–3):45–54CrossRefGoogle Scholar
  66. Špirić Z, Vučković I, Stafilov T, Kušan V, Frontasyeva M (2013) Air pollution study in Croatia using moss biomonitoring and ICP-AES and AAS analytical techniques. Arch Environ Contam Toxicol 65(1):33–46CrossRefGoogle Scholar
  67. Šprem N, Piria M, Barišić D, Kusak J, Barišić D (2016) Dietary items as possible sources of (137)Cs in large carnivores in the Gorski Kotar forest ecosystem, western Croatia. Sci Total Environ 542(Pt A):826–832Google Scholar
  68. Stoneberg RP, Jonkel CJ (1966) Age determination of black bears by cementum layers. J Wildl Manag 30:411–414CrossRefGoogle Scholar
  69. Thompson DR (1996) Mercury in birds and terrestrial mammals. In: Beyer WN, Heinz GH, Redmon-Norwood AW (eds) Environmental contaminants in wildlife: interpreting tissue residues. CRC, Boca Raton, pp 341–356Google Scholar
  70. Thompson DR, Stewart FM, Furness RW (1990) Using seabirds to monitor mercury in marine environments: the validity of conversion ratios for tissue comparisons. Mar Pollut Bull 21(7):339–342CrossRefGoogle Scholar
  71. Tsuji LJ, Wainman BC, Jayasinghe RK, VanSpronsen EP, Liberda EN (2009) Determining tissue-lead levels in large game mammals harvested with lead bullets: human health concerns. Bull Environ Contam Toxicol 82(4):435–439CrossRefGoogle Scholar
  72. Vahter M, Åkesson A, Lidén C, Ceccatelli S, Berglund M (2007) Gender differences in the disposition and toxicity of metals. Environ Res 104(1):85–95CrossRefGoogle Scholar
  73. Vesey DA (2010) Transport pathways for cadmium in the intestine and kidney proximal tubule: focus on the interaction with essential metals. Toxicol Lett 198:13–19CrossRefGoogle Scholar
  74. Vihnanek Lazarus M, Sekovanić A, Kljaković-Gašpić Z, Orct T, Jurasović J, Kusak J, Reljić S, Huber Đ (2013) Cadmium and lead in grey wolf liver samples: optimization of a microwave-assisted digestion method. Arch Ind Hyg Toxicol 64:395–403Google Scholar
  75. Wu X, Cobbina SJ, Mao G, Xu H, Zhang Z, Yang L (2016) A review of toxicity and mechanisms of individual and mixtures of heavy metals in the environment. Environ Sci Pollut Res Int 23(9):8244–8259CrossRefGoogle Scholar
  76. Yoshida M, Ohta H, Yamauchi Y, Seki Y, Sagi M, Yamazaki K, Sumi Y (1998) Age-dependent changes in metallothionein levels in liver and kidney of the Japanese. Biol Trace Elem Res 63:167–175CrossRefGoogle Scholar
  77. Žilinčár J, Zvada P, Kubín D, Hell P (1992) Die Schwermetallbelastung bei den Braunbären in den Westkarpaten. Z Jagdwiss 38:235–243Google Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Analytical Toxicology and Mineral Metabolism UnitInstitute for Medical Research and Occupational HealthZagrebCroatia
  2. 2.Department of Biology, Veterinary FacultyUniversity of ZagrebZagrebCroatia

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