Sizova EA, Yausheva EV, Shoshin DE, Nechitailo KS.

Animal Husbandry and Fodder Production. 2026. Vol. 109. No. 3. Р. 8-22.

doi: 10.33284/2658-3135-109-3-8

Original article

Biotoxicity of ultrafine zinc particles as a factor limiting their use in agriculture

 

Elena A Sizova1, Elena V Yausheva2, Daniil E Shoshin3, Ksenia S Nechitailo4

1,2,3,4Orenburg State University named after VA Bondarenko, Orenburg, Russia

1sizova.l78@yandex.ru, https://orcid.org/0000-0002-5125-5981

2vasilena56@mail.ru, https://orcid.org/0000-0002-1589-2211

3daniilshoshin@mail.ru, https://orcid.org/0000-0003-3086-681X

4k.nechit@mail.ru, https://orcid.org/0000-0002-8755-414X

Abstract. The widespread use of ultrafine metal particles (UFP) in agricultural practices, driven by their unique physicochemical properties, is accompanied by increasing environmental risks associated with their biotoxicity and bioaccumulation. Zinc is one of the most in-demand elements in agriculture, but the variability of the biological response of test samples depending on particle size and their colloidal behavior remains poorly understood, particularly in the submicron range, which exceeds the nanoscale. The aim of this study was to establish the relationship between the hydrodynamic diameter of ZnO UFP (458.41 ± 0.95 nm and 153.96 ± 2.57 nm) and their toxicity using two model systems: the bacterial strain Escherichia coli K12 TG1 and the ciliate Stylonychia mytilus. The results demonstrate pronounced size-dependent biotoxicity: particles with a smaller diameter (153.9 nm) exhibited a more prolonged inhibitory effect on bacterial luminescence over a wide range of low concentrations and caused more profound suppression of antioxidant enzymes (glutathione peroxidase, catalase, and superoxide dismutase) in ciliates compared to larger particles. At the same time, at the level of integrated indicators of survival and nucleic acid and protein content in S. mytilus, the size effect was neutralized, giving way to an overall toxic effect. It was found that even at subinhibitory concentrations, ZnO ultrafine particles induce critical oxidative stress, characterized by a threefold increase in glutathione concentration and an almost fivefold decrease in the activity of key antioxidant enzymes, indicating depletion of cellular defense mechanisms. Thus, despite the decrease in toxicity with increasing particle size, the high oxidative potential of zinc UFP remains a limiting factor for their widespread use in agriculture, justifying the need to develop compensatory strategies and regulations for safe use.

Keywords: ultrafine particles, Escherichia coli, Stylonychia mytilus, bioluminescence, oxidative stress, zinc

Acknowledgments: the research was carried out with the support of a grant for major scientific projects in priority areas of scientific and technological development (No. 075-15-2024-550).

For citation: Sizova EA, Yausheva EV, Shoshin DE, Nechitailo KS. Biotoxicity of ultrafine zinc particles as a factor limiting their use in agriculture. Animal Нusbandry and Fodder Production. 2026;109(3):8-22. (In Russ.). https://doi.org/10.33284/2658-3135-109-3-8

  1. Kostyuk VA, Potapovich AI, Kovaleva ZhV. A simple and sensitive method of determination of superoxide dismutase activity based on the reaction of quercetin oxidation. Questions of Medicinal Chemistry. 1990;36(2):88-91.
  2. 1.7.2.0018.15. Determination of nucleic acids in biological medicinal products using the Spirin method. Introduced 01.12.2018. M.: State Pharmacopoeia of the Russian Federation, XIV edition. 2018;2:1 р.
  3. Ayeleso TB, Ramachela K, Mukwevho E. Aqueous-methanol extracts of orange-fleshed sweet potato (Ipomoea batatas) ameliorate oxidative stress and modulate type 2 diabetes associated genes in insulin resistant C2C12 cells. Molecules. 2018;23(8):2058. doi: 10.3390/molecules23082058
  4. Dai H, Sun T, Han T, Guo Z, Wang X, Chen Y. Aggregation behavior of zinc oxide nanoparticles and their biotoxicity to Daphnia magna: Influence of humic acid and sodium alginate. Environ Res. 2020;191:110086. doi: 10.1016/j.envres.2020.110086
  5. Dai S, Ye R, Huang J, Wang B, Xie Z, Ou X, et al. Distinct lipid membrane interaction and uptake of differentially charged nanoplastics in bacteria. J Nanobiotechnology. 2022;20:191. doi: 10.1186/s12951-022-01321-z
  6. Dong X, Wu Z, Li X, Xiao L, Yang M, Li Y, et al. The size-dependent cytotoxicity of amorphous silica nanoparticles: a systematic review of in vitro studies. Int J Nanomed. 2020;15:9089-9113. doi: 10.2147/IJN.S276105
  7. Fatima A, Zaheer T, Pal K, Abbas RZ, Akhtar T, Ali S, et al. Zinc oxide nanoparticles significant role in poultry and novel toxicological mechanisms. Biol Trace Elem Res. 2024;202(1):268-290. doi: 10.1007/s12011-023-03651-x
  8. Fenchel T. Suspension feeding in ciliated protozoa: Functional response and particle size selection. Microb Ecol. 1980;6(1):1-11. doi: 10.1007/BF02020370
  9. Fröhlich E. The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles. Int J Nanomed. 2012;7:5577-5591. doi: 10.2147/IJN.S36111
  10. Garncarek-Musiał M, Dziewulska K, Kowalska-Góralska M. Effect of different sizes of nanocopper particles on rainbow trout (Oncorhynchus mykiss W.) spermatozoa motility kinematics. Sci Total Environ. 2024;941:173763. doi: 10.1016/j.scitotenv.2024.173763
  11. Gharpure S, Ankamwar B. Synthesis and Antimicrobial Properties of Zinc Oxide Nanoparticles. J Nanosci Nanotechnol. 2020;20(10):5977-5996. doi: 10.1166/jnn.2020.18707
  12. Hadwan MH, Abed HN. Data supporting the spectrophotometric method for the estimation of catalase activity. Data Brief. 2016;6:194-199. doi: 10.1016/j.dib.2015.12.002
  13. Heck DE, Shakarjian M, Kim HD, Laskin JD, Vetrano AM. Mechanisms of oxidant generation by catalase. Ann N Y Acad Sci. 2010;1203:120-125. doi: 10.1111/j.1749-6632.2010.05603.x
  14. Islam S. Toxicity and transport of nanoparticles in agriculture: effects of size, coating, and aging. Front Nanotechnol. 2025;7:1622228. doi: 10.3389/fnano.2025.1622228
  15. Kang M, Bai X, Liu Y, Weng Y, Wang H, Ye Z. Driving role of zinc oxide nanoparticles with different sizes and hydrophobicity in metabolic response and eco-corona formation in sprouts (Vigna radiata). Environ Sci Technol. 2024;58(22):9875-9886. doi: 10.1021/acs.est.4c01731
  16. Kumar S, Suman S, Muthukumar M, Mishra K, Bajpai A, Tiwari AK, et al. Technical review on different metal nanoparticles and their formulations on growth, agronomic and economic traits of crop plants. Front Nanotechnol. 2025;7:1576582. doi: 10.3389/fnano.2025.1576582
  17. Lavagna E, Barnoud J, Rossi G, Monticelli L. Size-dependent aggregation of hydrophobic nanoparticles in lipid membranes. 2020;12(17):9452-9461. doi: 10.1039/d0nr00868k
  18. Li J, Schiavo S, Rametta G, Miglietta ML, La Ferrara V, Wu C, et al. Comparative toxicity of nano ZnO and bulk ZnO towards marine algae Tetraselmis suecica and Phaeodactylum tricornutum. Environ Sci Pollut Res Int. 2017;24(7):6543-6553. doi: 10.1007/s11356-016-8343-0
  19. Lin X, Li J, Ma S, Liu G, Yang K, Tong M, et al. Toxicity of TiO2 nanoparticles to Escherichia coli: effects  of  particle  size,  crystal  phase and water chemistry. PLoS One. 2014;9(10):e110247. doi: 10.1371/journal.pone.0110247
  20. Makabenta JMV, Nabawy A, Li CH, Schmidt-Malan S, Patel R, Rotello VM. Nanomaterial-based therapeutics for  antibiotic-resistant  bacterial    Nat  Rev  Microbiol. 2021;19:23-36. doi: 10.1038/s41579-020-0420-1
  21. Muzammil S, Ashraf A, Siddique MH, Aslam B, Rasul I, Abbas R et al. A review on toxicity of nanomaterials in agriculture: Current scenario and future prospects. Sci Prog. 2023;106(4):368504231221672. doi: 10.1177/00368504231221672
  22. Nair S, Sasidharan A, Divya Rani VV, et al. Role of size scale of ZnO nanoparticles and microparticles on toxicity toward bacteria and osteoblast cancer cells. J Mater Sci Mater Med. 2009;20(Suppl 1):235-241. doi: 10.1007/s10856-008-3548-5
  23. Saliani M, Jalal R, Goharshadi EK Mechanism of oxidative stress involved in the toxicity of ZnO nanoparticles against eukaryotic cells. Nanomed J. 2016;3(1):1-14. doi: 10.7508/NMJ.2016.01.001
  24. Sattar AA, Matin AA, Hadwan MH, Hadwan AM, Mohammed RM. Rapid and effective protocol to measure glutathione peroxidase activity. Bull Natl Res Cent. 2024;48(1):100. doi: 10.1186/s42269-024-01250-x
  25. Schatzman SS, Peterson RL, Teka M, et al. Copper-only superoxide dismutase enzymes and iron starvation stress in Candida fungal pathogens. J Biol Chem. 2020;295(2):570-583. doi:10.1074/jbc.RA119.011084
  26. Singh R, Cheng S, Singh S. Oxidative stress-mediated genotoxic effect of zinc oxide nanoparticles on Deinococcus radiodurans. 3 Biotech. 2020;10(2):66. doi: 10.1007/s13205-020-2054-4
  27. Wang X, Shi Q, Zha Z, Zhu D, Zheng L, Shi L, et al. Copper single-atom catalysts with photothermal performance and enhanced nanozyme activity for bacteria‐infected wound therapy. Bioact Mater. 2021;6(12):4389-4401. doi: 10.1016/j.bioactmat.2021.04.024
  28. Zarghami A, Ganjkhanlou M, Zali A, Fekri A, Palangi V. Effects of nano-zinc oxide supplementation on milk yield, rumen fermentation, nutrient digestibility, and blood indices of high-yielding dairy cows. Front Vet Sci. 2025;12:1720270. doi: 10.3389/fvets.2025.1720270

Information about the authors:

Elena A Sizova, Dr. Sci. (Biology), professor, professor of the Russian Academy of Sciences, Chief Researcher Scientific in Educational Center «Biological Systems and Nanotechnology», Orenburg State University named after VA Bondarenko, 13 Pobedy Ave., Orenburg, 460018.

Elena V Yausheva, Cand. Sci. (Biology), Senior Researcher Scientific and Educational Center «Biological Systems and Nanotechnology», Orenburg State University named after VA Bondarenko, 13 Pobedy Ave., Orenburg, 460018.

Daniil E Shoshin, assistant Scientific and Educational Center «Biological Systems and Nanotechnology», Orenburg State University named after VA Bondarenko, 13 Pobedy Ave., Orenburg, 460018.

Ksenia S Nechitailo, Cand. Sci. (Biology), senior lecturer Scientific and Educational Center «Biological Systems and Nanotechnology», Orenburg State University named after VA Bondarenko, 13 Pobedy Ave., Orenburg, 460018.

The article was submitted 18.05.2026; approved after reviewing 07.07.2026; accepted for publication 14.09.2026.

Download