Environmental Science and Pollution Research

, Volume 25, Issue 16, pp 15765–15773 | Cite as

Assessment of biotoxicity of Cu nanoparticles with respect to probiotic strains of microorganisms and representatives of the normal flora of the intestine of broiler chickens

  • Aleksey Nikolayevich Sizentsov
  • Olga Vilorievna Kvan
  • Elena Petrovna Miroshnikova
  • Irina Aleksandrovna Gavrish
  • Victoria Alekseevna Serdaeva
  • Artem Vladimirovich Bykov
Research Article


Copper nanoparticle Cu (d = 55 ± 15 nm) and CuO nanoparticles (d = 90 ± 10 nm) were used in the studies (OOO Platina, Russia). Using the method of pure cultures, we extracted Lactobacillus, Enterococcus, and Enterobacterium from the intestines of broilers. Additionally, strains of Bacillus subtilis 10641 and Bifidobacterium were involved in probiotic strains. The data obtained in the course of the study testify to the insignificant biotoxicity of copper nanoparticles with respect to representatives of the genera Lactobacillus (30 to 15 μg/ml) and Bifidobacterium (30 μg/ml), with the most sensitive bacteria being the genus Lactobacillus, for which a concentration of 7.5 μg/ml was subinhibitory. The second stage was the study using method of agar wells. In the course of the experiment, we obtained results confirming the data of the research by the serial dilution method. In this case, as in the first case, the data indicate the insignificant biotoxicity of copper nanoparticles in relation to representatives of the genera Lactobacillus and Bifidobacterium. We have studied the bioaccumulating ability of microorganisms of the studied metals. In all the studies carried out, as in the first series of experiments, representatives of the genera Lactobacillus and Bifidobacterium with the lowest bioaccumulative ability were the most sensitive to copper nanoparticles and were 3.1 and 8.2%, respectively. The use of nanoparticles as a component of the fodder additive in small concentrations does not adversely affect not only the probiotic strains, but also the main representatives of the normoflora (Lactobacillus, Enterococcus, and Enterobacterium) of the poultry, the positive effect of the copper nanoparticles being directly related to low level of dissociation of nanoparticles, since biologically active ions will be released much more slowly, thereby creating a prolonged effect of exposure.


Cu nanoparticles Probiotics Bioaccumulation Lactobacillus Enterococcus Enterobacterium 


Funding information

This research was carried out with the financial support of the Russian Ministry of Science and Education as part of the base part of the state task to carry out research projects in the Orenburg State University (No. 6.6811.2017/Ch).

Compliance with ethical standards

The studies were carried out under the conditions of the experimental and biological clinic of the Orenburg State University on broiler chickens (Smena 8). Experimental studies were conducted in accordance with the instructions recommended by the Russian Regulations, 1987, and “The Guide for the Care and Use of Laboratory Animals (National Academy Press Washington, D.C. 1996).”


  1. Ahmadi F, Ebrahimnezhad Y, Sis NM, Ghalehkandi JG (2013) The effects of zinc oxide nanoparticles on performance, digestive organs and serum lipid concentrations in broiler chickens during starter period. Int J Biosci 7(23):23–29. CrossRefGoogle Scholar
  2. Auffan, M, Flahaut, E, Thill A, Mouchet F, Carriere M, Gauthier L, Bottero JY (2011) Ecotoxicology: nanoparticle reactivity and living organisms. In Nanoethics and nanotoxicology, Springer Berlin Heidelberg, рр. 325–357Google Scholar
  3. Beveridge TJ, Murray RG (1980) Sites of metal deposition in the cell wall of Bacillus subtilis. J Bacteriol 141(2):876–887Google Scholar
  4. Bogoslovskaya OA, Sizova EA, Polyakova VS, Miroshnikov SA, Leipunsky IO, Olkhovskaya IP, Glushchenko NN (2009) Investigation of the safety of the introduction of copper nanoparticles with different physicochemical characteristics into the animal organism. Vestnik OGU 2:124–127Google Scholar
  5. Chen M, Yamamuro S, Farrell D, Majetich SA (2003) Gold-coated iron nanoparticles for biomedical applications. J Appl Phys 93(10):7551–7553Google Scholar
  6. Churilov GI, Ivanycheva Y, Ampleev LE, Nazarova A, Zheglov T, Polishchuk S (2009) Introduction to the diet of rabbits wikis grown using ultrafine cobalt powders. Krolikovodstvo i zverovodstvo 1:16–17Google Scholar
  7. Claudio DM, Rojasa R (2006) Copper accumulation by bacteria and transfer to scallop larvae. Mar Pollut Bull 52(3):293–300CrossRefGoogle Scholar
  8. Ezhkov AM, Yapparov AK, Motina TY, Yapparov IA, Yezhkov VO (2015) Quality of meat of broiler chickens using the feed additive of nanosized bentonite. Glavnyj zootekhnik 1:45–49Google Scholar
  9. Folmanis GE, Kovalenko LV (2006) Biologically active iron nanopowders. MGOU Publishing House, MoscowGoogle Scholar
  10. Fondevila M, Herrer R, Casallas MC, Abecia L, Ducha JJ (2009) Silver nanoparticles as potential antimicrobial additive for weaned pigs. Anim Feed Sci Technol 150:259–269. CrossRefGoogle Scholar
  11. Galagudza MM, Korolev DV, Sonin DL, Aleksandrov IV, Postnov VN, Papayan GV, Shlyakhto EV (2010) Passive directed delivery of drugs to the ischemic myocardium using silica nanoparticles. Rossijskie nanotekhnologii 11-12:125–130Google Scholar
  12. Glushchenko NN, Bogoslovskaya OA, Olkhovskaya IP (2006) Comparative toxicity of salts and metal nanoparticles and their biological effect. Proceedings of the Academy of Industrial Ecology 3:46–47Google Scholar
  13. Glushchenko NN, Bogoslovskaya OA, Baitukalov TA, Olkhovskaya IP (2008) Nanoparticles of metals in bioelementology. Mikroehlementy v medicine 1-2:52Google Scholar
  14. Gusev AI (2005) Nanomaterials, nanostructures, nanotechnologies. Moscow, FizmatlitGoogle Scholar
  15. Hussain N, Jaitley V, Florence AT (2001) Recent advances in the understanding of uptake of microparticles across the gastrointestinal lymphatics. Adv Drug Deliv Rev 50:107–142CrossRefGoogle Scholar
  16. Isajkina EY, Sizentsov AN, Bunina AU, Shablo AS, Ovsyannikova DC (2015) Study of the bioaccumulating ability of probiotic preparations for the intoxication of laboratory animals with copper. Izvestia ОGAU 1(51):147–149Google Scholar
  17. Kasemets K, Ivask A, Dubourguier HC, Kahru A (2009) Toxicity of nanoparticles of ZnO, CuO and TiO2 to yeast Saccharomyces cerevisiae. Toxicol in Vitro 23(6):1116–1122CrossRefGoogle Scholar
  18. Kaweeteerawat C, Chang CH, Roy KR, Liu R, Li R, Toso D, Zhou ZH (2015) Cu nanoparticles have different impacts in Escherichia coli and Lactobacillus brevis than their microsized and ionic analogues. ACS Nano 9(7):7215–7225CrossRefGoogle Scholar
  19. Kensova R, Blazkova I, Vaculovicova M, Milosavljevic V, Blazkova L, Hynek D, Kopel P, Novotna M, Zehnalek J, Pohanka M, Trnkova L, Adam V, Kizek R (2015) The effect of cadmium ions and cadmium nanoparticles on chicken embryos and evaluation of organ accumulation. Int J Electrochem Sci 10:3623–3634Google Scholar
  20. Kim JH, Cho H, Ryu SE, Choi MU (2000) Effects of metal ions on the activity of protein tyrosine phosphatase VHR: highly potent and reversible oxidative inactivation by Cu2+ ion. Arch Biochem Biophys 382(1):72–80CrossRefGoogle Scholar
  21. Koch AM, Reynolds F, Merkle HP, Weissleder R, Josephson L (2005) Transport of surface-modified nanoparticles through cell monolayers. Chem bio chem 6(2):337–345CrossRefGoogle Scholar
  22. Kvan OB, Rusakova EA, Sizentsov AN, Kataev VY (2014) Effect of probiotic drugs on the content of toxic elements in the body of laboratory animals. Vestnik OGU 6(167):64–66Google Scholar
  23. Liang X, Sun M, Li L, Qiao R, Chen K, Xiao Q, Xu F (2012) Preparation and antibacterial activities of polyaniline/Cu0.05Zn0.95O nanocomposites. Dalton Trans 41(9):2804–2811CrossRefGoogle Scholar
  24. Lin YE, Vidic RD, Stout JE, McCartney CA, Yu VL (1998) Inactivation of Mycobacterium avium by copper and silver ions. Water Res 32(7):1997–2000CrossRefGoogle Scholar
  25. Makarov DV (2014) Forecast of the development of the world market of nanopowders. Vestnik KRAUNC Fiziko-matematicheskie nauki 1:97–102Google Scholar
  26. Maynard AD (2006) Nanotechnology: a research strategy for addressing risk. Woodrow Wilson International Center for Scholars, Washington, DC, USAGoogle Scholar
  27. Mroczek-Sosnowska N, Łukasiewicz M, Wnuk A, Sawosz E, Niemiec J, Skot A, Jaworski S, Chwalibog A (2016) In ovo administration of copper nanoparticles and copper sulfate positively influences chicken performance. J Sci Food Agric 96:3058–3062. CrossRefGoogle Scholar
  28. Niazi JH, Gu MB (2009) Toxicity of metallic nanoparticles in microorganism—a review. In Atmospheric and biological environmental monitoring, Ed. Y. J. Kim, Ed. Springer Science+Business MediaGoogle Scholar
  29. Ognik K, Sembratowicz I, Cholewińska E, Wlazło Ł, Nowakowicz-Dębek B, Szlązak R, Tutaj K (2016) The effect of chemically-synthesized silver nanoparticles on performance and the histology and microbiological profile of the jejunum in chickens. Ann Anim Sci 16:439–450. Google Scholar
  30. Peshkov SA, Sizentsov AN, Nikiyan AN, Kobzev GI (2015) Study of the bioaccumulation of heavy metals by bacteria of the genus Bacillus using X-ray fluorescence analysis and atomic force microscopy. Sovremennye problemy nauki i obrazovanija 4:50–53Google Scholar
  31. Pineda LM, Chwalibog A, Sawosz E, Lauridsen C, Engberg RM, Elnif J, Hotowy A, Sawosz F, Ali A, Gao Y, Moghaddam HS (2012a) Effect of silver nanoparticles on growth performance, metabolism and microbial profile of broiler chickens. Arch Anim Nutr 66:416–429. CrossRefGoogle Scholar
  32. Pineda L, Sawosz E, Hotowy A, Elnif J, Sawosz F, Ali A, Chwalibog A (2012b) Effect of nanoparticles of silver and gold on metabolic rate and development of broiler and layer embryos. Comp Biochem Physiol A Mol Integr Physiol 161(3):315–319Google Scholar
  33. Presnyak AR (2015) The use of micronutrient nanoparticles is a promising direction in the production of chicken broilers. Molodoj uchenyj 5(2):40–42Google Scholar
  34. Refaie AM, Ghazal MN, Easa FM, Barakat SA, Ge Y, Wh E (2015) Nano-copper as a new growth promoter in the diet of growing New Zealand white rabbits. Egypt J Rabbit Sci 25(1):39–57Google Scholar
  35. Rohner F, Ernst FO, Arnold M, Hilbe M, Biebinger R, Ehrensperger F, Pratsinis SE, Langhans W, Hurrell RF, Zimmermann MB (2007) Synthesis, characterization, and bioavailability in rats of ferric phosphate nanoparticles. J Nutr 137(3):614–619CrossRefGoogle Scholar
  36. Ruparelia JP, Chatterjee AK, Duttagupta SP, Mukherji S (2008) Strain specificity in antimicrobial activity of silver and copper nanoparticles. Acta Biomater 4(3):707–716CrossRefGoogle Scholar
  37. Sahoo SK, Labhasetwar V (2003) Nanotech approaches to drug delivery and imaging. Drug Discov Today 73:1112–1120. CrossRefGoogle Scholar
  38. Santos A, Mauro MS, Diaz DM (2006) Prebiotics and their long-term influence on the microbial populations of the mouse bowel. Food Microbiol 23:498–503. CrossRefGoogle Scholar
  39. Sawosz E, Binek M, Grodzik M, Zieliska M, Sysa P, Szmidt M, Niemiec T, Chwalibog A (2007) Influence of hydrocolloidal silver nanoparticles on gastrointestinal microflora and morphology of enterocytes of quails. Arch Anim Nutr 61:444–451. CrossRefGoogle Scholar
  40. Schwegmann H, Frimmel FH (2010) Nanoparticles: interaction with microorganisms. In: Frimmel FH, Niessner R (eds) Nanoparticles in the water cycle. Springer, Berlin, GermanyGoogle Scholar
  41. Sizentsov AN, Barysheva ES, Babushkina AE (2015) The ability of probiotic preparations based on bacteria of the genus Bacillus to bioaccumulate heavy metal ions in the body of laboratory animals. Russ Immunol J 9(2):753–755Google Scholar
  42. Sizentsov AN, Kvan OV, Babushkina АЕ, Исайкина Isajkina EY (2016) Bioaccumulative ability of bacteria of the genus Bacillus against lead ions under in vitro and in vivo conditions. Izvestia ОGAU 2(58): С.186–188Google Scholar
  43. Sizova EA (2010) Mineral composition and morphofunctional aspects of liver reorganization in the enteral method of introducing copper nanoparticles such as CU10X. Vestnik Orenburgskogo gosudarstvennogo universiteta 6:92–94Google Scholar
  44. Sizova EA, Korolev VL, Makaev SA, Miroshnikova EP, Shakhov VA (2016) Morpho-biochemical indicators of blood in broilers when correcting the diet with salts and nanoparticles Cu. Sel'skohozyajstvennaya biologiya 6:903–911CrossRefGoogle Scholar
  45. Stohs SJ, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med 18:321–336. CrossRefGoogle Scholar
  46. Taylor AA, Marcus IM, Guysi RL, Walker SL (2015) Metal oxide nanoparticles induce minimal phenotypic changes in a model colon gut microbiota. Environ Eng Sci 32:602–612. CrossRefGoogle Scholar
  47. Tsao N, Luh TY, Chou CK, Chang TY, Wu JJ, Liu CC, Lei HY (2002) In vitro action of carboxyfullerene. J Antimicrob Chemother 49(4):641–649CrossRefGoogle Scholar
  48. Vishnyakov AI, Ushakov AS, Lebedev SV (2011) Features of bone marrow hematopoiesis when copper nanoparticles are introduced per os and intramuscularly. Vestnik myasnogo skotovodstva 54:96–102Google Scholar
  49. Wang Z, Li N, Zhao J, White JC, Qu P, Xing B (2012) CuO nanoparticle interaction with human epithelial cells: cellular uptake, location, export, and genotoxicity. Chem Res Toxicol 25(7):1512–1521CrossRefGoogle Scholar
  50. Wang С, Wang MQ, Ye SS, Tao WJ, Du YJ, Wang C (2011) Effects of copper-loaded chitosan nanoparticles on growth and immunity in broilers. Poult Sci 90(10):2223–2228. CrossRefGoogle Scholar
  51. Yapparov AH, Aliev SA, Yapparov IA, Ezhkova AM, Degtyareva IA, Yezhkov VO (2014) Scientific substantiation of obtaining nanostructured and nanocomposite materials and technology of their use in agriculture. Kazan': Centr innovacionnyh tekhnologijGoogle Scholar
  52. Yausheva EV, Miroshnikov SA, Kosyan DB, Sizova ЕА (2016) Nanoparticles in combination with amino acids change productive and immunological indicators of broiler chicken. Agric Biol 51:912–920. Google Scholar
  53. Yausheva EV, Miroshnikov SA, Kvan OV (2013a) On the understanding of the biological effect of metal nanoparticles. Voprosy biologicheskoj, medicinskoj i farmacevticheskoj himii 9:54–56Google Scholar
  54. Yausheva EV, Zelepukhin AG, Ryabov NI, Kvan OV, Ramensky VA, Zaveryukha AK, Sirazetdinov FK (2013b) Investigation of metal nanoparticles as a source of microelements for animals. Sovremennye problemy nauki i obrazovaniya 5:470Google Scholar
  55. Yoon K, Hoon Byeon J, Park JH, Hwang J (2007) Susceptibility constants of Escherichia coli and Bacillus subtilis to silver and copper nanoparticles. Sci Total Environ 373(2–3):572–575CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Aleksey Nikolayevich Sizentsov
    • 1
  • Olga Vilorievna Kvan
    • 1
    • 2
  • Elena Petrovna Miroshnikova
    • 1
  • Irina Aleksandrovna Gavrish
    • 1
    • 2
  • Victoria Alekseevna Serdaeva
    • 2
  • Artem Vladimirovich Bykov
    • 1
  1. 1.Federal State Budget Educational Institution of Higher EducationOrenburg State UniversityOrenburgRussia
  2. 2.Federal State Budget Scientific InstitutionFederal Scientific Centre of Biological Systems and AgrotechnologiesOrenburgRussia

Personalised recommendations