Effects of food-borne ZnO nanoparticles on intestinal microbiota of common carp (Cyprinus carpio L.)

  • Latifeh ChupaniEmail author
  • Jiri Barta
  • Eliska Zuskova
Research Article


Ingestion of nanoparticles (NPs) with antimicrobial properties may disrupt the balance of intestinal microbiota. To investigate the effects of zinc oxide (ZnO) NPs on intestinal flora, common carp Cyprinus carpio were fed a commercial feed containing 500 mg kg−1 ZnO NPs for 6 weeks and compared to a control group receiving a similar feed-only regime. Sequencing data were analyzed both in individual fish and in pooled samples. Sequencing of 16S rRNA encoding gene of individual specimens revealed high variation in intestinal microbial composition. Assessment of pooled results can obscure high individual variation in data. ZnO NPs consumption was not associated with a significant difference in the intestinal microbial community compared to untreated controls. Our results indicated a high individual variation in the intestinal microbiome, which may further point out the importance of functional study over microbial composition to address nanomaterials-microbiome relationship.


ZnO Nanoparticles Microbiome Common carp Intestine 



We thank Sarah M. Owens and Stephanie Moormann Greenwald, Biosciences Division, Argonne National Laboratory, USA, for assistance in sequencing analysis. Our appreciation is extended to the Lucidus Consultancy, for the English edition of the manuscript.

Funding information

This research was supported by the Ministry of Education, Youth, and Sports of the Czech Republic projects CENAKVA (LM2018099) and the project Sustainable production of healthy fish in various aquaculture systems; PROFISH (CZ.02.1.01/0.0/0.0/16 019 /0000869).

Supplementary material

11356_2019_5616_MOESM1_ESM.docx (624 kb)
Online Resource 1 Alpha diversity analysis. Rarefaction curves of (A) Chao 1 index, (B) Observed OTUs (DOCX 623 kb)
11356_2019_5616_MOESM2_ESM.xlsx (24 kb)
ESM 2 (XLSX 23 kb)
11356_2019_5616_MOESM3_ESM.xlsx (134 kb)
ESM 3 (XLSX 133 kb)


  1. Adamovsky O, Buerger AN, Wormington AM, Ector N, Griffitt RJ, Bisesi Jr JH, Martyniuk CJ (2018) The gut microbiome and aquatic toxicology: An emerging concept for environmental health. Environ Toxicol Chem 37:2758–2775.
  2. Alonso VR, Guarner F (2013) Linking the gut microbiota to human health. Br J Nutr 109:S21–S26. CrossRefGoogle Scholar
  3. Beegam A, Prasad P, Jose J, Oliveira M, Costa FG, Soares AM, Gonçalves PP, Trindade T, Kalarikkal N, Thomas S (2016) Environmental fate of zinc oxide nanoparticles: risks and benefits. In: Soloneski S, Larramendy ML (eds) Toxicology - New Aspects to This Scientific Conundrum. InTech, Rijeka Croatia, pp 81–112Google Scholar
  4. Ben-Slama I, Mrad I, Rihane N, Mir LE, Sakly M, Amara S (2015) Sub-acute oral toxicity of zinc oxide nanoparticles in male rats. J Nanomed Nanotechnol 6:284. Google Scholar
  5. Burke C, Steinberg P, Rusch D, Kjelleberg S, Thomas T (2011) Bacterial community assembly based on functional genes rather than species. Proc Natl Acad Sci 108:14288–14293Google Scholar
  6. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7(5):335–336. CrossRefGoogle Scholar
  7. Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Lozupone CA, Turnbaugh PJ, Fierer N, Knight R (2011) Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. PNAS 108:4516–4522. CrossRefGoogle Scholar
  8. Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N, Owens SM, Betley J, Fraser L, Bauer M (2012) Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J 6:1621–1624. CrossRefGoogle Scholar
  9. Chupani L, Zusková E, Niksirat H, Panáček A, Lünsmann V, Haange SB, 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. CrossRefGoogle Scholar
  10. Chupani L, Niksirat H, Velisek J, Stara A, Hradilová S, Kolarik J, Panacek A, Zuskova E (2018a) Chronic dietary toxicity of zinc oxide nanoparticles in common carp (Cyprinus carpio L.): tissue accumulation and physiological responses. Ecotoxicol Environ Saf 147:110–116. CrossRefGoogle Scholar
  11. Chupani L, Niksirat H, Lünsmann V, Haange SB, von Bergen M, Jehmlich N, Zuskova E (2018b) Insight into the modulation of intestinal proteome of juvenile common carp (Cyprinus carpio L.) after dietary exposure to ZnO nanoparticles. Sci Total Environ 613:62–71. CrossRefGoogle Scholar
  12. Croteau M-N, Dybowska AD, Luoma SN, Valsami-Jones E (2011) A novel approach reveals that zinc oxide nanoparticles are bioavailable and toxic after dietary exposures. Nanotoxicology 5:79–90. CrossRefGoogle Scholar
  13. Edgar RC (2013) UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods 10:996–998. CrossRefGoogle Scholar
  14. Eichmiller JJ, Hamilton MJ, Staley C, Sadowsky MJ, Sorensen PW (2016) Environment shapes the fecal microbiome of invasive carp species. Microbiome 4:44. CrossRefGoogle Scholar
  15. Fröhlich EE, Fröhlich E (2016) Cytotoxicity of nanoparticles contained in food on intestinal cells and the gut microbiota. Int J Mol Sci:17–509.
  16. Janda JM, Abbott SL (2010) The genus Aeromonas: taxonomy, pathogenicity, and infection. Clin Microbiol Rev 23:35–73. CrossRefGoogle Scholar
  17. Jones N, Ray B, Ranjit KT, Manna AC (2008) Antibacterial activity of ZnO nanoparticle suspensions on a broad spectrum of microorganisms. FEMS Microbiol Lett 279:71–76. CrossRefGoogle Scholar
  18. Li T, Long M, Gatesoupe FJ, Zhang Q, Li A, Gong X (2015) Comparative analysis of the intestinal bacterial communities in different species of carp by pyrosequencing. Microb Ecol 69:25–36. CrossRefGoogle Scholar
  19. Loch TP, Faisal M (2015) Emerging flavobacterial infections in fish: a review. J Adv Res 6:283–300. CrossRefGoogle Scholar
  20. Londono Zuluaga N (2016) Impacts of engineered nanoparticles (TiO2, ZnO, Ag) on aquatic microbial communities.
  21. Merrifield DL, Shaw BJ, Harper GM, Saoud IP, Davies SJ, Handy RD, Henry TB (2013) Ingestion of metal-nanoparticle contaminated food disrupts endogenous microbiota in zebrafish (Danio rerio). Environ Pollut 174:157–163. CrossRefGoogle Scholar
  22. Nayak SK (2010) Role of gastrointestinal microbiota in fish. Aquac Res 41:1553–1573. CrossRefGoogle Scholar
  23. Shaw BJ, Handy RD (2011) Physiological effects of nanoparticles on fish: a comparison of nanometals versus metal ions. Environ Int. 37:1083–1097.
  24. Shen Z, Chen Z, Hou Z, Li T, Lu X (2015) Ecotoxicological effect of zinc oxide nanoparticles on soil microorganisms. FESE 9:912–918. Google Scholar
  25. Star B, Haverkamp TH, Jentoft S, Jakobsen KS (2013) Next generation sequencing shows high variation of the intestinal microbial species composition in Atlantic cod caught at a single location. BMC Microbiol 13:248. CrossRefGoogle Scholar
  26. Vaseem M, Umar A, Hahn YB (2010) ZnO nanoparticles: growth, properties, and applications. Metal oxide nanostructures and their applications. In: Umar A, Hahn YB (eds) Metal Oxide Nanostructures and Their Applications, vol 2010. American Scientific Publisher, New York, pp 1–36Google Scholar
  27. Wu S, Wang G, Angert ER, Wang W, Li W, Zou H (2012) Composition, diversity, and origin of the bacterial community in grass carp intestine. PLoS One 7:e30440. CrossRefGoogle Scholar
  28. Xie Y, He Y, Irwin PL, Jin T, Shi X (2011) Antibacterial activity and mechanism of action of zinc oxide nanoparticles against Campylobacter jejuni. Appl Environ Microbiol 77:2325–2331. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Fisheries and Protection of Waters, South Bohemian Research Centre of Aquaculture and Biodiversity of HydrocenosesUniversity of South Bohemia in České BudějoviceVodňanyCzech Republic
  2. 2.Department of Ecosystem Biology, Faculty of ScienceUniversity of South Bohemia in České BudějoviceČeské BudějoviceCzech Republic

Personalised recommendations