BACKGROUND: The high doses of zinc oxide (ZnO) administered orally to piglets for the prevention of diarrhea and increase of growth rate can contaminate pig farms and the surrounding environment. Therefore, there is a need to find a replacement of high doses of dietary ZnO with an equally effective alternative. In the present study, the effect of two formulations of zinc phosphate-based nanoparticles (ZnA and ZnC NPs) on growth performance, intestinal microbiota, antioxidant status, and intestinal and liver morphology was evaluated. A total of 100 weaned piglets were randomly divided into 10 equal groups with the base diet (control) or the base diet supplemented with ZnA, ZnC, or ZnO at concentrations 500, 1000, and 2000 mg Zn per kilogram of diet. Supplements were given to animals for 10 days. Fecal samples were collected on day 0, 5, 10 and 20. At the end of the treatment (day 10), three piglets from each group were sacrificed and analyzed. RESULTS: Comparing to that of control, the significantly higher piglet weight gain was observed in all piglet groups fed with ZnA (P < 0.05). Differences in the total aerobic bacteria and coliform counts in piglet feces after NPs supplementation compared to that of control and ZnO groups were also found (P < 0.05). The majority of aerobic culturable bacteria from the feces represented Escherichia (28.57-47.62%), Enterococcus (3.85-35.71%), and Streptococcus (3.70-42.31%) spp. A total of 542 Escherichia coli isolates were screened for the virulence genes STa, STb, Stx2, F4, and F18. The substantial occurrence of E. coli virulence factors was found on day 5, mainly in fimbrillary antigen and thermostable toxins, except for piglets fed by ZnC. Zn treatment decreased Zn blood levels in piglets fed with ZnO and ZnA (500 mg/kg) and increased in ZnC (2000 mg/kg) compared to that of control (P < 0.05). The antioxidant status of piglets was affected only by ZnA. While some changes in the liver and the intestinal morphology of piglets with NPs were observed, none were serious as reflected by the normal health status and increased weigh gain performance. CONCLUSIONS: Our results indicate that ZnA NPs have a positive effect on the piglet growth performance even at the lowest concentration. The prevalence of E. coli virulence factors was lowest in pigs supplemented with ZnC. Zinc phosphate-based nanoparticles may be an effective alternative to ZnO.

Central European Institute of Technology Brno University of Technology Purkynova 123 CZ 612 00 Brno Czech Republic

Central European Institute of Technology Center for Zoonoses University of Veterinary and Pharmaceutical Sciences Brno Palackeho 1946 1 CZ 612 42 Brno Czech Republic

Department of Animal Breeding Mendel University in Brno Zemedelska 1 CZ 613 00 Brno Czech Republic

Department of Animal Nutrition and Forage Production Mendel University in Brno Zemedelska 1 CZ 613 00 Brno Czech Republic

Department of Chemistry and Biochemistry Mendel University in Brno Zemedelska 1 CZ 613 00 Brno Czech Republic

Department of Inorganic Chemistry Faculty of Science Palacky University 17 listopadu 12 CZ 771 46 Olomouc Czech Republic

Institute of Animal Science Pratelstvi 815 CZ 104 00 Praha Uhrineves Czech Republic

Zobrazit více v PubMed

Poulsen HD. Zinc oxide for weanling piglets. Acta Agric Scand Sect A-Anim Sci. 1995;45(3):159–167.

Hu CH, Song J, You ZT, Luan ZS, Li WF. Zinc oxide-Montmorillonite hybrid influences diarrhea, intestinal mucosal integrity, and digestive enzyme activity in weaned pigs. Biol Trace Elem Res. 2012;149(2):190–196. PubMed

SCVMP . Standing Committee on Veterinary Medicinal Products (SCVMP): Summary report of the 19 June 2017 of the Standing Committee on Veterinary Medicinal Products. 2017.

EMA . European Medicines Agency (EMA): Committee for Medicinal Products for Veterinary Use (CVMP); Article 35. 2017.

NRC . National Research Council 2012: Nutrient requirements of swine: eleventh revised edition. Washington, DC: The National Academies Press; 2012.

Chi F, Johnston S, Tang X, Chen W, Wang B, Tang S. Effects of replacing zinc oxide and antibiotics with NeoPrime (R) on growth performance and plasma and fecal endotoxin concentration in nursery pigs. J Anim Sci. 2018;96:321.

Lei XJ, Kim IH. Low dose of coated zinc oxide is as effective as pharmacological zinc oxide in promoting growth performance, reducing fecal scores, and improving nutrient digestibility and intestinal morphology in weaned pigs. Anim Feed Sci Technol. 2018;245:117–125.

Jensen J, Larsen MM, Bak J. National monitoring study in Denmark finds increased and critical levels of copper and zinc in arable soils fertilized with pig slurry. Environ Pollut. 2016;214:334–340. PubMed

Bednorz Carmen, Oelgeschläger Kathrin, Kinnemann Bianca, Hartmann Susanne, Neumann Konrad, Pieper Robert, Bethe Astrid, Semmler Torsten, Tedin Karsten, Schierack Peter, Wieler Lothar H., Guenther Sebastian. The broader context of antibiotic resistance: Zinc feed supplementation of piglets increases the proportion of multi-resistant Escherichia coli in vivo. International Journal of Medical Microbiology. 2013;303(6-7):396–403. PubMed

Rensing C, Moodley A, Cavaco LM, McDevitt SF. Resistance to metals used in agricultural production. Microbiol Spectr. 2018;6(2):24. PubMed PMC

Horky P, Skalickova S, Urbankova L, Baholet D, Kociova S, Bytesnikova Z, et al. Zinc phosphate-based nanoparticles as a novel antibacterial agent: in vivo study on rats after dietary exposure. J Anim Sci Biotechnol. 2019;10:17. PubMed PMC

Pei X, Xiao ZP, Liu LJ, Wang G, Tao WJ, Wang MQ, et al. Effects of dietary zinc oxide nanoparticles supplementation on growth performance, zinc status, intestinal morphology, microflora population, and immune response in weaned pigs. J Sci Food Agric. 2019;99(3):1366–1374. PubMed

Wang C, Zhang LG, Ying ZX, He JT, Zhou L, Zhang LL, et al. Effects of dietary zinc oxide nanoparticles on growth, diarrhea, mineral deposition, intestinal morphology, and barrier of weaned piglets. Biol Trace Elem Res. 2018;185(2):364–374. PubMed

Jarvis TA, Miller RJ, Lenihan HS, Bielmyer GK. Toxicity of ZnO nanoparticles to the copepod Acartia tonsa, exposed through a phytoplankton diet. Environ Toxicol Chem. 2013;32(6):1264–1269. PubMed

Horky P, Skladanka J, Nevrkla P, Slama P. Effect of diet supplemented with antioxidants (selenium, copper, vitamins e and C) on antioxidant status and ejaculate quality of breeding boars. Ann Anim Sci. 2016;16(2):521–532.

Jancikova P, Horky P, Zeman L. The effect of feed additive containing vitamins and trace elements on the elements profile and growth of skin derivatives in horses. Ann Anim Sci. 2012;12(3):381–391.

Dzhumaniiazova IH, Khirazova EE, Bayzhumanov AA. Differences in effect of intermittent fasting on autonomic nervous system balance and antioxidant content in female and male Wistar rats. Acta Physiol. 2019;227:56.

Desai VG, Aidoo A, Li J, Lyn-Cook LE, Casciano DA, Feuers RJ. Effects of bleomycin on liver antioxidant enzymes and the electron transport system from ad libitum-fed and dietary-restricted female and male Fischer 344 rats. Nutr Cancer. 2000;36(1):42–51. PubMed

Mennecozzi M, Landesmann B, Palosaari T, Harris G, Whelan M. Sex differences in liver toxicity-do female and male human primary hepatocytes react differently to toxicants In Vitro? PLoS One. 2015;10(4):e0122786. PubMed PMC

Zurek L, Schal C. Evaluation of the German cockroach (Blattella germanica) as a vector for verotoxigenic Escherichia coli F18 in confined swine production. Vet Microbiol. 2004;101(4):263–267. PubMed

Lee SI, Kang SG, Kang ML, Yoo HS. Development of multiplex polymerase chain reaction assays for detecting enterotoxigenic Escherichia coli and their application to field isolates from piglets with diarrhea. J Vet Diagn Investig. 2008;20(4):492–496. PubMed

Meng JH, Zhao SH, Doyle MP. Virulence genes of Shiga toxin-producing Escherichia coli isolated from food, animals and humans. Int J Food Microbiol. 1998;45(3):229–235. PubMed

Bej AK, Mahbubani MH, Dicesare JL, Atlas RM. Polymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples. Appl Environ Microbiol. 1991;57(12):3529–3534. PubMed PMC

Barszcz M, Taciak M, Tusino A, Cobanova K, Gresakova L. The effect of organic and inorganic zinc source, used in combination with potato fiber, on growth, nutrient digestibility and biochemical blood profile in growing pigs. Livest Sci. 2019;227:37–43.

Zhao CY, Tan SX, Xiao XY, Qiu XS, Pan JQ, Tang ZX. Effects of dietary zinc oxide nanoparticles on growth performance and Antioxidative status in broilers. Biol Trace Elem Res. 2014;160(3):361–367. PubMed

Wang W, Van Noten N, Degroote J, Romeo A, Vermeir P, Michiels J. Effect of zinc oxide sources and dosages on gut microbiota and integrity of weaned piglets. J Anim Physiol Anim Nutr. 2019;103(1):231–241. PubMed

Fernandes CD, Resende M, Rodrigues LM, Garbossa CAP, Costa LB, Ferreira RA, et al. Dietary fiber and zinc additives on performance and intestinal health of Escherichia coli challenged piglets. Sci Agric. 2020;77(2):1–7.

Davin R, Manzanilla EG, Klasing KC, Perez JF. Effect of weaning and in-feed high doses of zinc oxide on zinc levels in different body compartments of piglets. J Anim Physiol Anim Nutr. 2013;97:6–12. PubMed

Li Y, Guo Y, Wen ZS, Jiang XM, Ma X, Han XY. Weaning stress perturbs gut microbiome and its metabolic profile in piglets. Sci Rep. 2018;8:12. PubMed PMC

Ding XH, Lan WS, Liu G, Ni HJ, Gu JD. Exploring possible associations of the intestine bacterial microbiome with the pre-weaned weight gaining performance of piglets in intensive pig production. Sci Rep. 2019;9:10. PubMed PMC

Pedersen RM, Gronnemose RB, Staerk K, Asferg CA, Andersen TB, Kolmos HJ, et al. A method for quantification of epithelium colonization capacity by pathogenic Bacteria. Front Cell Infect Microbiol. 2018;8:15. PubMed PMC

Otto M. Physical stress and bacterial colonization. FEMS Microbiol Rev. 2014;38(6):1250–1270. PubMed PMC

Jacobson M, Hård af Segerstad C, Gunnarsson A, Fellström C, de Verdier Klingenberg K, Wallgren P, Jensen-Waern M. Diarrhoea in the growing pig – a comparison of clinical, morphological and microbial findings between animals from good and poor performance herds. Research in Veterinary Science. 2003;74(2):163–169. PubMed PMC

Colombo G, Cortinovis C, Moschini E, Bellitto N, Perego MC, Albonico M, et al. Cytotoxic and proinflammatory responses induced by ZnO nanoparticles in in vitro intestinal barrier. J Appl Toxicol. 2019;39(8):1155–1163. PubMed

Bacchetta R, Moschini E, Santo N, Fascio U, Del Giacco L, Freddi S, et al. Evidence and uptake routes for zinc oxide nanoparticles through the gastrointestinal barrier in Xenopus laevis. Nanotoxicology. 2014;8(7):728–744. PubMed

Dubreuil JD, Isaacson RE, Schifferli DM. Animal Enterotoxigenic Escherichia coli. EcoSal Plus. 2016;7(1):1–47. PubMed PMC

Fairbrother JM, Nadeau E, Gyles CL. Escherichia coli in postweaning diarrhea in pigs: an update on bacterial types, pathogenesis, and prevention strategies. Anim Health Res Rev. 2005;6(1):17–39. PubMed

Soderlind O, Mollby R. Enterotoxins, O-groups, and K88 antigen in Escherichia coli from neonatal piglets with and without diarrhea. Infect Immun. 1979;24(3):611–616. PubMed PMC

Wilson RA, Francis DH. Fimbriae and enterotoxins associated with Escherichia coli serogroups isolated from pigs with colibacillosis. Am J Vet Res. 1986;47(2):213–217. PubMed

Zhang WP, Zhao MJ, Ruesch L, Omot A, Francis D. Prevalence of virulence genes in Escherichia coli strains recently isolated from young pigs with diarrhea in the US. Vet Microbiol. 2007;123(1–3):145–152. PubMed

Moredo FA, Pineyro PE, Marquez GC, Sanz M, Colello R, Etcheverria A, et al. Enterotoxigenic Escherichia coli subclinical infection in pigs: bacteriological and genotypic characterization and antimicrobial resistance profiles. Foodborne Pathog Dis. 2015;12(8):704–711. PubMed

Milani NC, Sbardella M, Ikeda NY, Arno A, Mascarenhas BC, Miyada VS. Dietary zinc oxide nanoparticles as growth promoter for weanling pigs. Anim Feed Sci Technol. 2017;227:13–23.

Wang Chao, Zhang Ligen, Su Weipeng, Ying Zhixiong, He Jintian, Zhang Lili, Zhong Xiang, Wang Tian. Zinc oxide nanoparticles as a substitute for zinc oxide or colistin sulfate: Effects on growth, serum enzymes, zinc deposition, intestinal morphology and epithelial barrier in weaned piglets. PLOS ONE. 2017;12(7):e0181136. PubMed PMC

Afifi Mohamed, Almaghrabi Omar A., Kadasa Naif Mohammed. Ameliorative Effect of Zinc Oxide Nanoparticles on Antioxidants and Sperm Characteristics in Streptozotocin-Induced Diabetic Rat Testes. BioMed Research International. 2015;2015:1–6. PubMed PMC

Bharathi D, Bhuvaneshwari V. Synthesis of zinc oxide nanoparticles (ZnO NPs) using pure bioflavonoid rutin and their biomedical applications: antibacterial, antioxidant and cytotoxic activities. Res Chem Intermed. 2019;45(4):2065–2078.

Gupta R, Malik P, Das N, Singh M. Antioxidant and physicochemical study of Psidium guajava prepared zinc oxide nanoparticles. J Mol Liq. 2019;275:749–767.

Liu JH, Ma X, Xu YY, Tang H, Yang ST, Yang YF, et al. Low toxicity and accumulation of zinc oxide nanoparticles in mice after 270-day consecutive dietary supplementation. Toxicol Res. 2017;6(2):134–143. PubMed PMC

Hosseindoust AR, Lee SH, Kim JS, Choi YH, Noh HS, Lee JH, et al. Dietary bacteriophages as an alternative for zinc oxide or organic acids to control diarrhoea and improve the performance of weanling piglets. Vet Med. 2017;62(2):53–61.

Kanikowska A, Wlochal M, Mielcarz G, Grzymislawski M, Kucharski M. Evaluation of zinc and copper concentrations and the total antioxidant capacity of blood plasma in patients with malabsorption syndrome. J Elem. 2015;20(4):873–885.

Bondzio Angelika, Pieper Robert, Gabler Christoph, Weise Christoph, Schulze Petra, Zentek Juergen, Einspanier Ralf. Feeding Low or Pharmacological Concentrations of Zinc Oxide Changes the Hepatic Proteome Profiles in Weaned Piglets. PLoS ONE. 2013;8(11):e81202. PubMed PMC

Long LN, Chen JS, Zhang YG, Liang X, Ni HJ, Zhang B, et al. Comparison of porous and nano zinc oxide for replacing high-dose dietary regular zinc oxide in weaning piglets. PLoS One. 2017;12(8):14. PubMed PMC

Xia T, Lai WQ, Han MM, Han M, Ma X, Zhang LY. Dietary ZnO nanoparticles alters intestinal microbiota and inflammation response in weaned piglets. Oncotarget. 2017;8(39):64878–64891. PubMed PMC

García-Gómez Concepción, García Sandra, Obrador Ana, Almendros Patricia, González Demetrio, Fernández María Dolores. Effect of ageing of bare and coated nanoparticles of zinc oxide applied to soil on the Zn behaviour and toxicity to fish cells due to transfer from soil to water bodies. Science of The Total Environment. 2020;706:135713. PubMed

Ameen F, AlYahya S, Govarthanan M, Aljahdali N, Al-Enazi N, Alsamhary K, et al. Soil bacteria Cupriavidus sp. mediates the extracellular synthesis of antibacterial silver nanoparticles. J Mol Struct. 2020;1202:1–6.

Zheng Min, Huang Zhen, Ji Haodong, Qiu Fuguo, Zhao Dongye, Bredar Alexandria R.C., Farnum Byron H. Simultaneous control of soil erosion and arsenic leaching at disturbed land using polyacrylamide modified magnetite nanoparticles. Science of The Total Environment. 2020;702:134997. PubMed

Josko I. Copper and zinc fractionation in soils treated with CuO and ZnO nanoparticles: the effect of soil type and moisture content. Sci Total Environ. 2019;653:822–832. PubMed

In vivo evaluation of selenium-tellurium based nanoparticles as a novel treatment for bovine mastitis

Metabolic adaptations of Escherichia coli to extended zinc exposure: insights into tricarboxylic acid cycle and trehalose synthesis

Zinc effects on bacteria: insights from Escherichia coli by multi-omics approach

Is a new generation of mycotoxin clay adsorbents safe in a pig's diet?

Importance of Zinc Nanoparticles for the Intestinal Microbiome of Weaned Piglets

Protective effect of a new generation of activated and purified bentonite in combination with yeast and phytogenic substances on mycotoxin challenge in pigs