Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Multiorgan histopathological changes in the juvenile seabream Sparus aurata as a biomarker for zinc oxide particles toxicity

  • 140 Accesses

  • 2 Citations


Zinc oxide nanoparticles are widely used in some domains (cosmetics, pharmaceuticals optical devices, and agricultural field) due to their physical, optical, and antimicrobial properties. However, the release of ZnO-NPs into the environment may affect organisms like fish with potential consequences for human health. Histological approaches of the acute effects of these materials on fish are scarce; thus, the present study aimed to investigate the potential toxic effects of acute exposure to ZnO particles in marine environments, by assessing histological changes in the gills, liver, spleen, and muscle of gilthead seabream (Sparus aurata) juveniles. Thus, fish were exposed for 96 h, via water, to 1 mg L−1 of ionic zinc and zinc oxide particles (1.1, 1.2, and 1.4 μm of size). Histological examination revealed gills as the most affected organ, followed by liver, muscle, and spleen. In the gills, histopathological changes included hyperplasia of epithelial cells, fusion of the secondary lamellae, and lifting of the lamellar epithelium with edema. In the liver, lipid vacuolation of several degrees, necrosis of hepatic and pancreatic tissues, blood congestion in sinusoids and hepatoportal vessels, presence of cellular infiltrate, and melano-macrophages diffusion was found. Muscle showed degeneration, atrophy, thickening and necrosis of muscle fibers with edema between them, and presence of melano-macrophages in the muscle layer. Spleen was the less damaged organ, displaying congested blood, white pulp increase/rupture, and bigger and darker melano-macrophage aggregates in the splenic stroma. These results underline that the size of particles plays a determinant role in their potential pernicious effects. A short-term exposure caused major histopathological changes in relevant organs of S. aurata juveniles, possibly affecting their function.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Abbas HH, Ali FK (2007) Study the effect of hexavalent chromium on some biochemical, citotoxicological and histopathological aspects of the Orechromis spp. fish. Pak J Biol Sci 10:3973–3982

  2. Abdel-Khalek AA, Kadry M, Hamed A, Marie M-A (2015) Ecotoxicological impacts of zinc metal in comparison to its nanoparticles in Nile tilapia; Oreochromis niloticus. J Basic Appl Zool 72:113–125

  3. Afifi M, Alkaladi A, Abu-Zinada O, Zaki ZN (2014) Histopathological and ultrastractural alterations in skin, gills, liver and muscle of Siganus canaliculatus and Epinephelus morio caught from Jeddah and Yanbu coast as bio-indicators of oil hydrocarbons pollution. Life Sci J Life Sci J 11(8)

  4. Agius C, Roberts RJ (2003) Melano-macrophage centres and their role in fish pathology. J Fish Dis 26:499–509

  5. Al-Bairuty GA, Shaw BJ, Handy RD, Henry TB (2013) Histopathological effects of waterborne copper nanoparticles and copper sulphate on the organs of rainbow trout (Oncorhynchus mykiss). Aquat Toxicol 126:104–115

  6. Altinok I, Capkin E (2007) Histopathology of rainbow trout exposed to sublethal concentrations of methiocarb or endosulfan. Toxicol Pathol 35:405–410

  7. Au DW (2004) The application of histo-cytopathological biomarkers in marine pollution monitoring: a review. Mar Pollut Bull 48:817–834

  8. Bacaksiz E, Parlak M, Tomakin M, Ozcelik A, Karakiz M, Altunbas M (2008) The effect of zinc nitrate, zinc acetate and zinc chloride precursors on investigation of structural and optical properties of ZnO thin films. J Alloy Compd 466:447–450

  9. Barreto Â, Luis LG, Girão AV, Trindade T, Soares AMVM, Oliveira M (2015) Behavior of colloidal gold nanoparticles in different ionic strength media. J Nanopart Res J Nanopart Res 17:1–13

  10. Beegam A, Prasad P, Jose J, Oliveira M, Costa FG, Soares AMVM, et al (2016) Environmental fate of zinc oxide nanoparticles: risks and benefits, In: Toxicology - New Aspects to This Scientific Conundrum pp, 81–112, Inthec Open

  11. Caballero MJ, Izquierdo MS, Kjorsvik E, Fernandez AJ, Rosenlund G (2004) Histological alterations in the liver of sea bream, Sparus aurata L., caused by short- or long-term feeding with vegetable oils. Recovery of normal morphology after feeding fish oil as the sole lipid source. J Fish Dis 27:531–541

  12. Camargo MMP, Martinez CBR (2007) Histopathology of gills, kidney and liver of a neotropical fish caged in an urban stream Neotropical Ichthyology. Neotrop Ichthyol 5:327–336

  13. Chupani L, Zusková E, Niksirat H, Panáček A, Lünsmann V, Haange S-B, von Bergen M, Jehmlich N (2017) Effects of chronic dietary exposure of zinc oxide nanoparticles on the serum protein profile of juvenile common carp (Cyprinus carpio L.). Sci Total Environ 579:1504–1511

  14. David M, Kartheek RM (2015) Histopathological alterations in spleen of freshwater fish Cyprinus carpio exposed to sublethal concentration of sodium cyanide. Open Vet J 5:1–5

  15. DelValls TA, Blasco J, Sarasquete MC, Forja JM, Gomez-Parra A (1998) Evaluation of heavy metal sediment toxicity in littoral ecosystems using juveniles of the fish Sparus aurata. Ecotoxicol Environ Saf 41:157–167

  16. Dzika E, Rotkiewicz T, Hoffmann RW (2005) Histopathological examination of liver and spleen of roach Rutilus rutilus (L.) originating from selected lakes of Warmia and Mazury Lakeland in Poland. Zool Poloniae 50:33–47

  17. Evans DH, Piermarini PM, Choe KP (2005) The multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid-base regulation, and excretion of nitrogenous waste. Physiol Rev 85:97–177

  18. Figueiredo-Fernandes A, Ferreira-Cardoso JV, Garcia-Santos S, Monteiro SM, Carrola J, Matos P, Fontaínhas-Fernandes A (2007) Histopathological changes in liver and gill epithelium of Nile tilapia, Oreochromis niloticus, exposed to waterborne copper. Pesq Vet Bras 27:103–109

  19. Flores-Lopes F, Thomaz A (2011) Histopathologic alterations observed in fish gills as a tool in environmental monitoring. J Braz J Biol 71:179–188

  20. Gaber HS, Abbas W, Authman M, Gaber SA (2014) Histological and biochemical studies on some organs of two fish species in Bardawil Lagoon, North Sinai, Egypt. Glob Vet 12(1):1–11

  21. Gonzalez De Canales ML, Oliva M, Garrido C (2009) Toxicity of lindane (gamma-hexachloroxiclohexane) in Sparus aurata, Crassostrea angulata and Scrobicularia plana. J Environ Sci Health B 44:95–105

  22. Hadi AA, Alwan SF (2012) Histopathological changes in gills, liver and kidney of fresh water fish, Tilapia zillii, exposed to aluminum. Int J Pharm Life Sci 3(11):2071–2081

  23. Jiang J, Pi J, Cai J (2018) The advancing of zinc oxide nanoparticles for biomedical applications. J Bioinorg Chem Appl 2018:18

  24. Kaoud HA, El-Dahshan AR (2010) Bioaccumulation and histopathological alterations of the heavy metals in Oreochromis niloticus fish. Nat Sci 8:147–156

  25. Khosravi-Katuli K, Lofrano G, Pak Nezhad H, Giorgio A, Guida M, Aliberti F, Siciliano A, Carotenuto M, Galdiero E, Rahimi E, Libralato G (2018) Effects of ZnO nanoparticles in the Caspian roach (Rutilus rutilus caspicus). Sci Total Environ 626:30–41

  26. Kurtović B, Teskeredzic E, Teskeredžic Z (2008) Histological comparison of spleen and kidney tissue from farmed and wild European sea bass (Dicentrarchus labrax L.). Acta Adriat 49(2):147–154

  27. Mansouri B, Maleki A, Davari B, Johari SA, Shahmoradi B, Mohammadi E, Shahsavari S (2016) Histopathological effects following short-term coexposure of Cyprinus carpio to nanoparticles of TiO2 and CuO. Environ Monit Assess 188:575

  28. Mansouri B, Johari SA, Azadi A, Sarkheil M (2018) Effects of Waterborne ZnO Nanoparticles and Zn2+ ions on the gills of rainbow trout (Oncorhynchus mykiss): bioaccumulation, histopathological and ultrastructural changes. Turk J Fish Aquat Sci 18:739–746

  29. Masini MA, Prato P, Giacco E, Mariottini G, Pane L (2014) Impact of synthetic surfactants and oil dispersants on the gills of juvenile gilthead (Sparus aurata L.). J Biol Res 87:2296

  30. Mohamed ASF (2009) Histopathological studies on Tilapia zillii and Solea vulgaris from Lake Qarun, Egypt. World J Fish Marine Sci 1:29–39

  31. Montero D, Blazer VS, Socorro J, Izquierdo MS, Tort L (1999) Dietary and culture influences on macrophage aggregate parameters in gilthead seabream (Sparus aurata) juveniles. Aquaculture 179:523–534

  32. Oliveira M, Maria VL, Ahmad I, Serafim A, Bebianno MJ, Pacheco M, Santos MA (2009) Contamination assessment of a coastal lagoon (Ria de Aveiro, Portugal) using defence and damage biochemical indicators in gill of Liza aurata - an integrated biomarker approach. Environ Pollut 157:959–967

  33. Ostaszewska T, Chojnacki M, Kamaszewski M, Sawosz-Chwalibog E (2016) Histopathological effects of silver and copper nanoparticles on the epidermis, gills, and liver of Siberian sturgeon. Environ Sci Pollut Res Int 23:1621–1633

  34. Patnaik BB, Howrelia JH, Mathews T, Selvanayagam M (2011) Histopathology of gill, liver, muscle and brain of Cyprinus carpio communis L. exposed to sublethal concentration of lead and cadmium. Afr J Biotechnol 10(57):12218–12223

  35. Rajkumar KS, Kanipandian N, Thirumurugan R (2016) Toxicity assessment on haemotology, biochemical and histopathological alterations of silver nanoparticles-exposed freshwater fish Labeo rohita. Appl Nanosci 6:19–29

  36. Rudnicki CAM, Melo GC, Donatti L, Kawall HG, Fanta E (2009) Gills of juvenile fish piaractus mesopotamicus as histological biomarkers for experimental sub-lethal contamination with the Organophosphorus Azodrin®400. Braz Arch Biol Technol 52:1431–1441

  37. Saad SMM, El-Deeb AE, Tayel SI, Al-Shehri E, Ahmed NAM (2012) Effect of heavy metals pollution on histopathological alterations in muscles of Clarias gariepinus inhabiting the Rosetta branch, River Nile, Egypt. In: 1st International conference of biotechnology applied agriculture Benha University, Egypt, pp 79–88

  38. Sabir S, Arshad M, Khalil S (2014) Zinc oxide nanoparticles for revolutionizing agriculture: synthesis and applications. Sci World J 2014:8

  39. Santos D, Regina S, Melo M, Carla S, Mendes D, Karoline BS, Rocha I, Silva J, Cantanhêde S et al (2014) Histological changes in gills of two fish species as indicators of water quality in Jjansen lagoon (São Luís, Maranhão State, Brazil). Int J Environ Res Public Health 11(12):12927–12937

  40. Sayed AE-D, Abdel Fatah M Younes H (2017) Melanomacrophage centers in Clarias gariepinus as immunological biomarker for toxicity of silver nanoparticles. J Microsc Ultrastruct

  41. Shahi N, Sarma D, Pandey J, Das P, Sarma D, Mallik SK (2016) Impact of acid mine drainage on haematological, histopathological and genotoxic effects in golden mahaseer, Tor putitora. J Environ Biol 37:509–515

  42. Shaw B, Handy R (2011) Physiological effects of nanoparticles on fish: a comparison of nanometals versus metal ions. Environ Int 37(6):1083–1097

  43. Soo Choi J, Kim R-O, Yoon S, Kim W-K (2016) Developmental toxicity of zinc oxide nanoparticles to zebrafish (Danio rerio): a transcriptomic analysis. PLoS One 11(8):e0160763

  44. Subashkumar S, Selvanayagam M (2014) First report on: acute toxicity and gill histopathology of fresh water fish cyprinus Cyprinus carpio exposed to zinc oxide (zno) nanoparticles. Int J Sci Res Publ 4:10–13

  45. Sun T, Gottschalk F, Hungerbuhler K, Nowack B (2014) Comprehensive probabilistic modelling of environmental emissions of engineered nanomaterials. Environ Pollut 185:69–76

  46. Teles M, Fierro-Castro C, Na-Phatthalung P, Tvarijonaviciute A, Soares AMVM, Tort L, Oliveira M (2016) Evaluation of gemfibrozil effects on a marine fish (Sparus aurata) combining gene expression with conventional endocrine and biochemical endpoints. J Hazard Mater 318:600–607

  47. Vergilio CD, Carvalho C, Melo E (2012) Accumulation and histopathological effects of mercury chloride after acute exposure in tropical fish Gymnotus carapo. J Chem Health Risks 2(4):01–08

  48. Wolf JC, Wolfe MC (2005) A brief overview of nonneoplastic hepatic toxicity in fish. Toxicol Pathol 33:75–85

  49. Yung MMN, Mouneyrac C, Leung KMY (2014) Ecotoxicity of zinc oxide nanoparticles in the marine environment. In: Bhushan B (ed) Encyclopedia of Nanotechnology. Springer Netherlands, Dordrecht, pp 1–17

  50. Zhang Y, Nayak TR, Hong H, Cai W (2013) Biomedical applications of zinc oxide nanomaterials. Curr Mol Med 13:1633–1645

  51. Zhao X, Wang S, Wu Y, You H, Lv L (2013) Acute ZnO nanoparticles exposure induces developmental toxicity, oxidative stress and DNA damage in embryo-larval zebrafish. Aquat Toxicol 136-137:49–59

  52. Zhong Q, Matijević E (1996) Preparation of uniform zinc oxide colloids by controlled double-jet precipitation. J Mater Chem 6:443–447

Download references

Conflict of interest

The authors declare that they have no conflicts of interest.


Funds were received by Project CICECO-Aveiro Institute of Materials, FCT Ref. UID/CTM/50011/2019, financed by national funds through the FCT/MCTES and BIC/UI189/7424/2016. Thanks are due for the financial support to CESAM (UID/AMB/50017/2019), to FCT/MCTES through national funds. MO had financial support of the program Investigator FCT (IF/00335/2015), co-funded by the Human Potential Operational Programme and European Social Fund.

Author information

Correspondence to Miguel Oliveira.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Responsible editor: Cinta Porte

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Beegam, A., Lopes, M., Fernandes, T. et al. Multiorgan histopathological changes in the juvenile seabream Sparus aurata as a biomarker for zinc oxide particles toxicity. Environ Sci Pollut Res (2019). https://doi.org/10.1007/s11356-019-05949-7

Download citation


  • Zinc oxide particles
  • Fish
  • Biomarkers
  • Histopathology
  • Acute exposure