FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Front Vet Sci / Monitoring trace minerals and heavy metals in liver of free-...
Frontiers in veterinary science2026; 13; 1751586; doi: 10.3389/fvets.2026.1751586

Monitoring trace minerals and heavy metals in liver of free-living large herbivores in the Netherlands.

Abstract: Trace minerals are essential for animal health but can also, together with heavy metals, have a negative impact, making their monitoring crucial to assess animal health. These elements were examined through a long-term post-mortem monitoring system based on routine liver sampling for Heck cattle, Konik horses and red deer in place at the Oostvaardersplassen nature reserve in the Netherlands, using data from this system to determine reference intervals and investigate trends in liver trace element concentrations. Throughout the monitoring programme, inductively coupled plasma mass spectrometry was used to measure concentrations of trace minerals and heavy metals, including arsenic, cadmium, chromium, cobalt, copper, iron, lead, manganese, molybdenum, nickel, selenium, vanadium, and zinc. Species-specific patterns in trace element profiles were identified, with red deer showing comparatively higher copper levels and horses elevated iron and lead levels. Temporal declines in certain elements, including iron and lead, were observed across all species. Seasonal and age-related variations were also evident. Importantly, reference intervals estimated in this study differed from livestock standards, in particular for copper and selenium, highlighting the need for species- and context-specific reference intervals when assessing health in free-living herbivores. These findings provide valuable baseline data for ongoing environmental and health monitoring in minimally managed, multi-species populations at the reserve, highlighting the importance of mineral surveillance in free-living animals to enhance wildlife health assessment, track long-term environmental changes, and support management decisions in nature reserves across the Netherlands and more globally.
Copyright © 2026 Marcelino, Monti, Cornelissen, Bassingthwaighte, het Lam, van der Merwe and van der Poel.
Get Full Text
Publication Date: 2026-02-24 PubMed ID: 41815500PubMed Central: PMC12971517DOI: 10.3389/fvets.2026.1751586Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This study monitored trace minerals and heavy metals in the livers of free-living large herbivores in the Oostvaardersplassen nature reserve in the Netherlands.
  • The research provided species-specific reference values and revealed temporal trends, emphasizing the importance of tailored mineral monitoring for wildlife health assessment and environmental management.

Background and Importance

  • Trace minerals, such as copper and selenium, are essential nutrients for animal health, supporting physiological functions.
  • Heavy metals, like lead and cadmium, though sometimes naturally present, can accumulate and pose toxicity risks to animals.
  • Monitoring these elements helps assess the health status of free-ranging animals and detect environmental contamination or nutritional imbalances.
  • Free-living large herbivores like Heck cattle, Konik horses, and red deer can serve as bioindicators of the ecosystem’s mineral status.

Study Design and Methods

  • A long-term post-mortem monitoring system was established at Oostvaardersplassen, a nature reserve in the Netherlands, involving routine liver sampling of three herbivore species: Heck cattle, Konik horses, and red deer.
  • Liver tissue was selected because it is a key organ for mineral storage and reflects animal mineral status.
  • Concentrations of multiple trace elements and heavy metals were measured using inductively coupled plasma mass spectrometry (ICP-MS), a sensitive and precise analytical technique.
  • The elements analyzed included arsenic, cadmium, chromium, cobalt, copper, iron, lead, manganese, molybdenum, nickel, selenium, vanadium, and zinc.

Key Findings

  • Species-specific mineral profiles:
    • Red deer livers showed relatively higher copper concentrations compared to Konik horses and Heck cattle.
    • Konik horses exhibited elevated iron and lead levels compared to the other species.
  • Temporal trends:
    • There was a decline in liver concentrations of certain elements, specifically iron and lead, observed in all species over time.
    • These trends may reflect changes in environmental exposure or dietary sources across the years.
  • Seasonal and age-related variations:
    • Levels of trace elements fluctuated according to the season, possibly linked to variations in food availability or physiological demands.
    • Age-dependent differences were found, indicating that mineral metabolism or accumulation changes during animal development or aging.
  • Reference intervals:
    • The study established species- and population-specific reference intervals for trace minerals and heavy metals in liver tissue.
    • These reference values differed noticeably from livestock standards commonly used for domestic animals, especially for copper and selenium.
    • This highlights the importance of using tailored reference values when assessing free-living wildlife, as domestic animal standards may not be appropriate.

Implications and Applications

  • The data serve as an important baseline for ongoing health and environmental monitoring of minimally managed wildlife populations.
  • Monitoring liver mineral concentrations aids in early detection of deficiencies or toxicities that could affect animal health and population viability.
  • Tracking long-term changes in trace element levels helps identify shifts in environmental contamination or nutrient cycling within the reserve.
  • The findings support informed management decisions for the nature reserve to maintain ecosystem health and biodiversity.
  • The methodology and reference intervals can potentially be applied to other free-living herbivore populations in the Netherlands and internationally.

Conclusion

  • This study underscores the necessity of species- and context-specific mineral monitoring programs in wildlife conservation.
  • By establishing detailed baseline data and identifying temporal and biological variation patterns, it enhances understanding of mineral status in free-ranging herbivores.
  • Such monitoring is critical for wildlife health assessment, environmental surveillance, and effective ecosystem management in nature reserves.

Cite This Article

APA
Marcelino I, Monti G, Cornelissen P, Bassingthwaighte E, Het Lam J, van der Merwe D, van der Poel WHM. (2026). Monitoring trace minerals and heavy metals in liver of free-living large herbivores in the Netherlands. Front Vet Sci, 13, 1751586. https://doi.org/10.3389/fvets.2026.1751586

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 13
Pages: 1751586
PII: 1751586

Researcher Affiliations

Marcelino, Inês
  • Infectious Disease Epidemiology, Wageningen University and Research, Wageningen, Netherlands.
Monti, Gustavo
  • Infectious Disease Epidemiology, Wageningen University and Research, Wageningen, Netherlands.
Cornelissen, Perry
  • Department Nature and Society, Staatsbosbeheer, Lelystad, Netherlands.
  • Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, Netherlands.
Bassingthwaighte, Evelyn
  • School of Veterinary Science, University of Queensland, Gatton, QLD, Australia.
Het Lam, Jasper
  • Department of Ruminant Health, Royal GD, Deventer, Netherlands.
van der Merwe, Deon
  • Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK, United States.
  • Department of Research and Development, Royal GD, Deventer, Netherlands.
van der Poel, Wim H M
  • Infectious Disease Epidemiology, Wageningen University and Research, Wageningen, Netherlands.
  • Department of Virology and Molecular Biology, Wageningen Bioveterinary Research, Lelystad, Netherlands.

Conflict of Interest Statement

JL and DM were employed by Royal GD. The remaining author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References

This article includes 50 references
  1. Barboza PS, Parker KL, Hume ID. Metabolic constituents: water, minerals and vitamins. Integrative Wildlife Nutrition Berlin, Heidelberg: Springer; (2009). p. 157–206.
  2. Suttle NF. Mineral Nutrition of Livestock, 5th Edn. Wallingford, Oxfordshire: CAB International (2022).
  3. Geffré A, Friedrichs K, Harr K, Concordet D, Trumel C, Braun J-P. Reference values: a review. Vet Clin Pathol (2009) 38:288–98.
    doi: 10.1111/j.1939-165X.2009.00179.xpubmed: 19737162google scholar: lookup
  4. Friedrichs KR, Harr KE, Freeman KP, Szladovits B, Walton RM, Barnhart KF. American society for veterinary clinical pathology. ASVCP reference interval guidelines: determination of de novo reference intervals in veterinary species and other related topics. Vet Clin Pathol (2012) 41:441–53.
    doi: 10.1111/vcp.12006pubmed: 23240820google scholar: lookup
  5. Jota Baptista C, Seixas F, Gonzalo-Orden JM, Oliveira PA. Biomonitoring metals and metalloids in wild mammals: invasive versus non-invasive sampling. Environ Sci Pollut Res Int (2022) 29:18398–407.
    doi: 10.1007/s11356-022-18658-5pubmed: 35032272google scholar: lookup
  6. Puls R. Mineral Levels in Animal Health: Diagnostic Data, 2nd Edn. Clearbrook, BC: Sherpa International (1994).
  7. Moore AR, Camus MS, Harr K, Kjelgaard-Hansen M, Korchia J, Jeffery U. Systematic evaluation of 106 laboratory reference data articles from nondomestic species published from 2014 to 2016: assessing compliance with reference interval guidelines. J Zoo Wildl Med (2020) 51:469–77.
    doi: 10.1638/2019-0186pubmed: 33480521google scholar: lookup
  8. Gordon IJ, Hester AJ, Festa-Bianchet M. The management of wild large herbivores to meet economic, conservation and environmental objectives. J Appl Ecol (2004) 41:1021–31.
    doi: 10.1111/j.0021-8901.2004.00985.xgoogle scholar: lookup
  9. Barboza PS, Rombach EP, Blake JE, Nagy JA. Copper status of muskoxen: a comparison of wild and captive populations. J Wildl Dis (2003) 39:610–9.
    doi: 10.7589/0090-3558-39.3.610pubmed: 14567223google scholar: lookup
  10. Tajchman K, Ukalska-Jaruga A, Bogdaszewski M, Pecio M, Janiszewski P. Comparison of the accumulation of macro- and microelements in the bone marrow and bone of wild and farmed red deer (). BMC Vet Res (2021) 17:324.
    doi: 10.1186/s12917-021-03041-2pmc: PMC8502351pubmed: 34627246google scholar: lookup
  11. Draghi S, Spinelli M, Fontanarosa C, Curone G, Amoresano A, Pignoli E. Evaluation of the difference in the content of essential and non-essential elements in wild boar and swine tissues sampled in the same area of Northern Italy. Animals (2024) 14:827.
    doi: 10.3390/ani14060827pmc: PMC10967638pubmed: 38539924google scholar: lookup
  12. van der Merwe D, Jacobs Y, van der Drift S, Koopmans A. Reference intervals for trace minerals and heavy metals in livers of cattle in wilderness areas in the Netherlands. J Vet Diagn Invest (2025) 37:552–8.
    doi: 10.1177/10406387251325864pmc: PMC11948226pubmed: 40128632google scholar: lookup
  13. Hollingsworth KA, Shively RD, Glasscock SN, Light JE, Tolleson DR, Barboza PS. Trace mineral supplies for populations of little and large herbivores. PLoS ONE (2021) 16:e0248204.
    doi: 10.1371/journal.pone.0248204pmc: PMC7959371pubmed: 33720946google scholar: lookup
  14. Wenting E, Siepel H, Jansen PA. Variability of the Ionome of Wild Boar (Sus scrofa) and Red Deer () in a Dutch National Park, with Implications for biomonitoring. Biol Trace Elem Res (2024) 202:2518–46.
    doi: 10.1007/s12011-023-03879-7pmc: PMC11052835pubmed: 37814170google scholar: lookup
  15. Marcelino I, Keizer J, Monti G, Cornelissen P, Santman-Berends I, Lam JH. Monitoring pathogens in free-living large herbivores in a nature reserve in the Netherlands. Transbound Emerg Dis (2025) 2025:6948049.
    doi: 10.1155/tbed/6948049pmc: PMC12356683pubmed: 40822449google scholar: lookup
  16. Cornelissen P, Bokdam J, Sykora K, Berendse F. Effects of large herbivores on wood pasture dynamics in a European wetland system. Basic Appl Ecol (2014) 15:396–406.
    doi: 10.1016/j.baae.2014.06.006google scholar: lookup
  17. van Geel P, Poelmann P, van der List M. Advies Beheer Oostvaardersplassen. Kaders voor provinciaal beleid Provincie Flevoland. Flevoland: Externe Begeleidingscommissie beheer Oostvaardersplassen. (2018).
  18. Thrusfield MV, Christley R. Veterinary Epidemiology, Fourth Edn. Hoboken, NJ: Wiley Blackwell; (2018).
  19. Counotte G, Holzhauer M, Dijken SC, Muskens J, der Merwe DV. Levels of trace elements and potential toxic elements in bovine livers: a trend analysis from 2007 to 2018. PLoS ONE (2019) 14:e0214584.
    doi: 10.1371/journal.pone.0214584pmc: PMC6456170pubmed: 30964882google scholar: lookup
  20. van der Merwe D, van den Wollenberg L, van Hees-Valkenborg J, de Haan T, van der Drift S, Vandendriessche V. Evaluation of hair analysis for determination of trace mineral status and exposure to toxic heavy metals in horses in the Netherlands. J Vet Diagn Invest (2022) 34:1000–5.
    doi: 10.1177/10406387221116069pmc: PMC9597333pubmed: 35918902google scholar: lookup
  21. R Core Team. R: A Language and Environment for Statistical Computing. (2024).
  22. Wickham H, Averick M, Bryan J, Chang W, McGowan LD, François R. Welcome to the Tidyverse. J Open Source Softw (2019) 4:1686.
    doi: 10.21105/joss.01686google scholar: lookup
  23. Wickham H. ggplot2: Elegant Graphics for Data Analysis. New York: Springer International Publishing (2016).
  24. Oksanen J, Simpson G, Blanchet F, Kindt R, Legendre P, Minchin P. Vegan: Community Ecology Package. (2025).
  25. Hui W, Gel YR, Gastwirth JL. lawstat: An R package for law, public policy and biostatistics. J Stat Softw (2008) 28:1–26.
    doi: 10.18637/jss.v028.i03google scholar: lookup
  26. Davison AC, Hinkley DV. Bootstrap Methods and Their Application. Cambridge: Cambridge University Press; (1997).
  27. Finnegan D. referenceIntervals: Reference Intervals. (2024).
  28. Royal GD. Pakket zware metalen en mineralenscreening leverbiopt. Gddiergezondheid (2023).
  29. van der Merwe D, van den Wollenberg L, van Hees-Valkenborg J, de Haan T, van der Drift S. Reference intervals for trace mineral and heavy metal concentrations in horse livers in the Netherlands. J Vet Diagn Invest (2023) 35:737–41.
    doi: 10.1177/10406387231193328pmc: PMC10621559pubmed: 37565635google scholar: lookup
  30. Draghi S, Fehri NE, Ateş F, Özsobaci NP, Tarhan D, Bilgiç B. Use of hair as matrix for trace elements biomonitoring in cattle and roe deer sharing pastures in Northern Italy. Animals (2024) 14:2209.
    doi: 10.3390/ani14152209pmc: PMC11311060pubmed: 39123735google scholar: lookup
  31. Gupta RC. Veterinary Toxicology : Basic and Clinical Principles, 2nd Edn. Oxford: Academic (2012).
  32. Ramsay WN. Age-related storage of iron in the liver of horses. Vet Res Commun (1994) 18:261–8.
    doi: 10.1007/BF01839192pubmed: 7831755google scholar: lookup
  33. Kabata-Pendias A. Trace Elements in Soils and Plants, 4th Edn. Boca Raton: CRC Press (2011).
  34. Cornelissen P. Waterkwaliteit open water Oostvaardersplassen 29 April 2020. Intern Report Amersfoort: Staatsbosbeheer. (2020).
  35. French AS, Shaw D, Gibb SW, Taggart MA. Geochemical landscapes as drivers of trace and toxic element profiles in wild red deer (). Sci Total Environ (2017) 601–602:1606–18.
    doi: 10.1016/j.scitotenv.2017.05.210pubmed: 28609848google scholar: lookup
  36. European Environment Agency. Heavy metal emissions in Europe. (2025).
  37. Vikøren T, Bernhoft A, Waaler T, Handeland K. Liver concentrations of copper, cobalt, and selenium in wild Norwegian red deer (). J Wildl Dis (2005) 41:569–79.
    doi: 10.7589/0090-3558-41.3.569pubmed: 16244067google scholar: lookup
  38. Durkalec M, Nawrocka A, Krzysiak M, Larska M, Kmiecik M, Posyniak A. Trace elements in the liver of captive and free-ranging European bison ( L). Chemosphere (2018) 193:454–63.
    doi: 10.1016/j.chemosphere.2017.11.050pubmed: 29154121google scholar: lookup
  39. Wilson PR, Grace ND. A review of tissue reference values used to assess the trace element status of farmed red deer (). N Z Vet J (2001) 49:126–32.
    doi: 10.1080/00480169.2001.36219pubmed: 16032179google scholar: lookup
  40. Handeland K, Viljugrein H, Lierhagen S, Opland M, Tarpai A, Vikøren T. Low copper levels associated with low carcass weight in wild red deer () in Norway. J Wildl Dis (2017) 53:176–80.
    doi: 10.7589/2016-02-037pubmed: 27788057google scholar: lookup
  41. Pilarczyk B, Balicka-Ramisz A, Ramisz A, Adamowicz E, Pilarczyk R, Tomza-Marciniak A. Selenium concentration in liver and kidney of free living animals (roe and red deer) from West Pomerania (Poland). Eur J Wildl Res (2009) 55:279–83.
    doi: 10.1007/s10344-008-0247-ygoogle scholar: lookup
  42. Flueck WT, Smith-Flueck JM, Mionczynski J, Mincher BJ. The implications of selenium deficiency for wild herbivore conservation: a review. Eur J Wildl Res (2012) 58:761–80.
    doi: 10.1007/s10344-012-0645-zgoogle scholar: lookup
  43. Cammilleri G, Messina EMD, Pantano L, Buscemi MD, Migliore S, Blanda V. Trace metals and metalloids in wild boars (S. scrofa) from Southern Italy: comparison between urban and wild areas. Chemosphere (2025) 380:144473.
    doi: 10.1016/j.chemosphere.2025.144473pubmed: 40347669google scholar: lookup
  44. Vikøren T, Kristoffersen AB, Lierhagen S, Handeland K. A comparative study of hepatic trace element levels in wild moose, roe deer, and reindeer from Norway. J Wildl Dis (2011) 47:661–72.
    doi: 10.7589/0090-3558-47.3.661pubmed: 21719831google scholar: lookup
  45. Herrada A, Bariod L, Saïd S, Rey B, Bidault H, Bollet Y. Minor and trace element concentrations in roe deer hair: a non-invasive method to define reference values in wildlife. Ecol Indic (2024) 159:111720.
    doi: 10.1016/j.ecolind.2024.111720google scholar: lookup
  46. Zemanova MA. Towards more compassionate wildlife research through the 3Rs principles: moving from invasive to non-invasive methods. Wildlife Biol (2020) 2020:wlb.00607.
    doi: 10.2981/wlb.00607google scholar: lookup
  47. Bleich VC, Stewart KM. Geographic variation in trace mineral concentrations in blood of mule deer from the Mojave Desert, California, USA. California Fish Wildl J (2025) 111:e13.
    doi: 10.51492/cfwj.111.13google scholar: lookup
  48. Le Boedec K. Reference interval estimation of small sample sizes: a methodologic comparison using a computer-simulation study. Vet Clin Pathol (2019) 48:335–46.
    doi: 10.1111/vcp.12725pubmed: 31228287google scholar: lookup
  49. Ammer T, Schützenmeister A, Prokosch H-U, Rauh M, Rank CM, Zierk J. refineR: a novel algorithm for reference interval estimation from real-world data. Sci Rep (2021) 11:16023.
    doi: 10.1038/s41598-021-95301-2pmc: PMC8346497pubmed: 34362961google scholar: lookup
  50. Hoffmann G, Klawitter S, Trulson I, Adler J, Holdenrieder S, Klawonn F. Novel tool for the rapid and transparent verification of reference intervals in clinical laboratories. J Clin Med (2024) 13:4397.
    doi: 10.3390/jcm13154397pmc: PMC11313426pubmed: 39124664google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,000 horse owners like you who receive our email updates on horse health and nutrition.