In China’s swine industry, copper is generally supplemented above the National Research Council (NRC) requirement (2012) because of its antimicrobial properties and the potential for growth promotion. Yet few are concerned about whether this excess supplementation is necessary. In this study, the 16S rRNA pyrosequencing was designed and used to investigate the effect of dietary copper level on the diversity of the fecal microbial community and the correlation of copper level with the serum level of inflammatory cytokines in Sprague-Dawley rat models. The results showed that the diet containing a high level of Cu (120 and 240 mg/kg) changed the microbial richness and diversity of rat feces associated with the increased copper content in the rat ileac and colonic digesta. Furthermore, a Pearson’s correlation analysis indicated that an accumulation of unabsorbed copper in the chyme was correlated with the microbial composition of the rat feces, which was linked with TNF-α in serum. The results suggest that dietary copper level may have a direct impact on circulating inflammatory cytokines in the serum, perhaps inducing an inflammatory response by altering the microbial composition of rat feces. Serum TNF-α could be the chief responder to excessive copper exposure.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Amato, K.R., Yeoman, C.J., Kent, A., Righini, N., Carbonero, F., Estrada, A., Gaskins, H.R., Stumpf, R.M., Yildirim, S., Torralba, M., et al. 2013. Habitat degradation impacts black howler monkey (Alouatta pigra) gastrointestinal microbiomes. ISME J. 7, 1344–1353.
Ansteinsson, V., Refsnes, M., Skomedal, T., Osnes, J.B., Schiander, I., and Lag, M. 2009. Zinc- and Copper-induced interleukin-6 release in primary cell cultures from rat heart. Cardiovasc. Toxicol. 9, 86–94.
Armstrong, T.A., Cook, D.R., Ward, M.M., Williams, C.M., and Spears, J.W. 2004. Effect of dietary copper source (cupric citrate and,cupric sulfate) and concentration on growth performance and fecal copper excretion in weanling pigs. J. Anim. Sci. 82, 1234–1240.
Bailey, J.D., Ansotegui, R.P., Paterson, J.A., Swenson, C.K., and Johnson, A.B. 2001. Effects of supplementing combinations of inorganic and complexed copper on performance and liver mineral status of beef heifers consuming antagonists. J. Anim. Sci. 79, 2926–2934.
Bailey, M.T., Dowd, S.E., Galley, J.D., Hufnagle, A.R., Allen, R.G., and Lyte, M. 2011. Exposure to a social stressor alters the structure of the intestinal microbiota: Implications for stressor-induced immunomodulation. Brain Behav. Immun. 25, 397–407.
Boente, R.F., Ferreira, L.Q., Falcao, L.S., Miranda, K.R., Guimaraes, P.L.S., Santos, J., Vieira, J.M.B.D., Barroso, D.E., Emond, J.P., Ferreira, E.O., et al. 2010. Detection of resistance genes and susceptibility patterns in Bacteroides and Parabacteroides strains. Anaerobe 16, 190–194.
Dewar, M.L., Arnould, J.P., Dann, P., Trathan, P., Groscolas, R., and Smith, S. 2013. Interspecific variations in the gastrointestinal microbiota in penguins. Microbiologyopen 2, 195–204.
Dias, R.S., Lopez, S., Montanholi, Y.R., Smith, B., Haas, L.S., Miller, S.P., and France, J. 2013. A meta-analysis of the effects of dietary copper, molybdenum, and sulfur on plasma and liver copper, weight gain, and feed conversion in growing-finishing cattle. J. Anim. Sci. 91, 5714–5723.
Dziarski, R., Park, S.Y., Kashyap, D.R., Dowd, S.E., and Gupta, D. 2016. Pglyrp-regulated gut microflora Prevotella falsenii, Parabacteroides distasonis and Bacteroides eggerthii enhance and Alistipes finegoldii attenuates colitis in mice. PLoS One 11, 1–24.
Felske, A., Wolterink, A., van Lis, R., De Vos, W.M., and Akkermans, A.D.L. 1999. Searching for predominant soil bacteria: 16S rDNA cloning versus strain cultivation. FEMS Microbiol. Ecol. 30, 137–145.
Frank, D.N., Amand, A.L.S., Feldman, R.A., Boedeker, E.C., Harpaz, N., and Pace, N.R. 2007. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc. Natl. Acad. Sci. USA 104, 13780–13785.
Fry, R.S., Ashwell, M.S., Lloyd, K.E., O’Nan, A.T., Flowers, W.L., Stewart, K.R., and Spears, J.W. 2012. Amount and source of dietary copper affects small intestine morphology, duodenal lipid peroxidation, hepatic oxidative stress, and mRNA expression of hepatic copper regulatory proteins in weanling pigs. J. Anim. Sci. 90, 3112–3119.
Gong, J.H., Forster, R.J., Yu, H., Chambers, J.R., Sabour, P.M., Wheatcroft, R., and Chen, S. 2002. Diversity and phylogenetic analysis of bacteria in the mucosa of chicken ceca and comparison with bacteria in the cecal lumen. FEMS Microbiol. Lett. 208, 1–7.
Gowanlock, D.W., Mahan, D.C., Jolliff, J.S., Moeller, S.J., and Hill, G.M. 2013. Evaluating the NRC levels of Cu, Fe, Mn, and Zn using organic minerals for grower-finisher swine. J. Anim. Sci. 91, 5680–5686.
Hojberg, O., Canibe, N., Poulsen, H.D., Hedemann, M.S., and Jensen, B.B. 2005. Influence of dietary zinc oxide and copper sulfate on the gastrointestinal ecosystem in newly weaned piglets. Appl. Environ. Microbiol. 71, 2267–2277.
Huang, Y.L., Ashwell, M.S., Fry, R.S., Lloyd, K.E., Flowers, W.L., and Spears, J.W. 2015. Effect of dietary copper amount and source on copper metabolism and oxidative stress of weanling pigs in short-term feeding. J. Anim. Sci. 93, 2948–2955.
Jondreville, C., Revy, P.S., and Dourmad, J.Y. 2003. Dietary means to better control the environmental impact of copper and zinc by pigs from weaning to slaughter. Livest. Prod. Sci. 84, 147–156.
Jost, T., Lacroix, C., Braegger, C.P., Rochat, F., and Chassard, C. 2014. Vertical mother-neonate transfer of maternal gut bacteria via breastfeeding. Environ. Microbiol. 16, 2891–2904.
Konstantinov, S.R. 2005. Ph.D. thesis. Lactobacilli in the porcine intestine: from composition to functionality. Wageningen University, Wageningen, Netherlands.
Kumar, V., Kalita, J., Bora, H.K., and Misra, U.K. 2016. Relationship of antioxidant and oxidative stress markers in different organs following copper toxicity in a rat model. Toxicol. Appl. Pharmacol. 293, 37–43.
Lin, Z.M., Ning, H.F., Bi, J.G., Qiao, J.F., Liu, Z.H., Li, G.H., Wang, Q.S., Wang, S.H., and Ding, Y.F. 2014. Effects of nitrogen fertilization and genotype on rice grain macronutrients and micronutrients. Rice Science 21, 233–242.
Liu, J.H., Zhang, M.L., Zhang, R.Y., Zhu, W.Y., and Mao, S.Y. 2016. Comparative studies of the composition of bacterial microbiota associated with the ruminal content, ruminal epithelium and in the faeces of lactating dairy cows. Microb. Biotechnol. 9, 257–268.
Lu, L., Wang, R.L., Zhang, Z.J., Steward, F.A., Luo, X.G., and Liu, B. 2010. Effect of dietary supplementation with copper sulfate or tribasic copper chloride on the growth performance, liver copper concentrations of broilers fed in floor pens, and stabilities of vitamin E and phytase in feeds. Biol. Trace. Elem. Res. 138, 181–189.
Luo, X.G., Ji, F., Lin, Y.X., Steward, F.A., Lu, L., Liu, B., and Yu, S.X. 2005. Effects of dietary supplementation with copper sulfate or tribasic copper chloride on broiler performance, relative copper bioavailability, and oxidation stability of vitamin E in feed. Poult. Sci. 84, 888–893.
Ma, Y.L., Zanton, G.I., Zhao, J., Wedekind, K., Escobar, J., and Vazquez-Anon, M. 2015. Multitrial analysis of the effects of copper level and source on performance in nursery pigs. J. Anim. Sci. 93, 606–614.
Mattie, M.D., McElwee, M.K., and Freedman, J.H. 2008. Mechanism of copper-activated transcription: activation of AP-1, and the JNK/SAPK and p38 signal transduction pathways. J. Mol. Biol. 383, 1008–1018.
Mavromichalis, I., Hancock, J.D., Kim, I.H., Senne, B.W., Kropf, D.H., Kennedy, G.A., Hines, R.H., and Behnke, K.C. 1999. Effects of omitting vitamin and trace mineral premixes and(or) reducing inorganic phosphorus additions on growth performance, carcass characteristics, and muscle quality in finishing pigs. J. Anim. Sci. 77, 2700–2708.
Mei, S.F., Yu, B., Ju, C.F., Zhu, D., and Chen, D.W. 2010. Effect of different levels of copper on growth performance and cecal ecosystem of newly weaned piglets. Int. J. Mol. Sci. 9, 378–381.
Mori, H., Maruyama, F., Kato, H., Toyoda, A., Dozono, A., Ohtsubo, Y., Nagata, Y., Fujiyama, A., Tsuda, M., and Kurokawa, K. 2014. Design and experimental application of a novel non-degenerate universal primer set that amplifies prokaryotic 16S rRNA genes with a low possibility to amplify eukaryotic rRNA genes. DNA Res. 21, 217–227.
Munoz, C., Lopez, M., Olivares, M., Pizarro, F., Arredondo, M., and Araya, M. 2005. Differential response of interleukin-2 production to chronic copper supplementation in healthy humans. Eur. Cytokine Netw. 16, 261–265.
Namkung, H., Gong, J., Yu, H., and De Lange, C.F.M. 2006. Effect of pharmacological intakes of zinc and copper on growth performance, circulating cytokines and gut microbiota of newly weaned piglets challenged with coliform lipopolysaccharides. Can. J. Anim. Sci. 86, 511–522.
Novotny, J., Pistl, J., and Kovac, G. 2003. Effects of supplementation of organic-bound trace elements on blood and tissues-Micromineral profile and immune parameters of piglets. Acta Vet.-Beogr. 53, 11–18.
Pang, Y., Patterson, J.A., and Applegate, T.J. 2009. The influence of copper concentration and source on ileal microbiota. Poult. Sci. 88, 586–592.
Pereira, T.C., Campos, M.M., and Bogo, M.R. 2016. Copper toxicology, oxidative stress and inflammation using zebrafish as experimental model. J. Appl. Toxicol. 36, 876–881.
Petta, S., Gastaldelli, A., Rebelos, E., Bugianesi, E., Messa, P., Miele, L., Svegliati-Baroni, G., Valenti, L., and Bonino, F. 2016. Pathophysiology of non alcoholic fatty liver disease. Int. J. Mol. Sci. 17, 1–26.
Rajilic-Stojanovic, M., Shanahan, F., Guarner, F., and De Vos, W.M. 2013. Phylogenetic analysis of dysbiosis in ulcerative colitis during remission. Inflamm. Bowel Dis. 19, 481–488.
Reeves, P.G., Nielsen, F.H., and Fahey, G.C.Jr. 1993. AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. J. Nutr. 123, 1939–1951.
Sanchez, D., Miguel, M., and Aleixandre, A. 2012. Dietary fiber, gut peptides, and adipocytokines. J. Med. Food 15, 223–230.
Satokari, R., Fuentes, S., Mattila, E., Jalanka, J., De Vos, W.M., and Arkkila, P. 2014. Fecal transplantation treatment of antibioticinduced, noninfectious colitis and long-term microbiota followup. Case. Rep. Med. 2014, 1–7.
Shelton, J.L., Southern, L.L., LeMieux, F.M., Bidner, T.D., and Page, T.G. 2004. Effects of microbial phytase, low calcium and phosphorus, and removing the dietary trace mineral premix on carcass traits, pork quality, plasma metabolites, and tissue mineral content in growing-finishing pigs. J. Anim. Sci. 82, 2630–2639.
Singh, K.K., Kumar, M., Kumar, P., Gupta, M.K., Jha, D.K., Kumari, S., Roy, B.K., and Kumar, S. 2012. “Free” copper: a new endogenous chemical mediator of inflammation in birds. Biol. Trace. Elem. Res. 145, 338–348.
Song, J., Li, Y.L., and Hu, C.H. 2013. Effects of copper-exchanged montmorillonite, as alternative to antibiotic, on diarrhea, intestinal permeability and proinflammatory cytokine of weanling pigs. Appl. Clay Sci. 77-78, 52–55.
Turnlund, J.R., Jacob, R.A., Keen, C.L., Strain, J.J., Kelley, D.S., Domek, J.M., Keyes, W.R., Ensunsa, J.L., Lykkesfeldt, J., and Coulter, J. 2004. Long-term high copper intake: effects on indexes of copper status, antioxidant status, and immune function in young men. Am. J. Clin. Nutr. 79, 1037–1044.
Veum, T.L., Carlson, M.S., Wu, C.W., Bollinger, D.W., and Ellersieck, M.R. 2004. Copper proteinate in weanling pig diets for enhancing growth performance and reducing fecal copper excretion compared with copper sulfate. J. Anim. Sci. 82, 1062–1070.
Walter, R.M., Uriuhare, J.Y., Olin, K.L., Oster, M.H., Anawalt, B.D., Critchfield, J.W., and Keen, C.L. 1991. Copper, zinc, manganese, and magnesium status and complications of diabetes-mellitus. Diabetes Care 14, 1050–1056.
Wang, M.Q., Du, Y.J., Wang, C., Tao, W.J., He, Y.D., and Li, H. 2012. Effects of copper-loaded chitosan nanoparticles on intestinal microflora and morphology in weaned piglets. Biol. Trace. Elem. Res. 149, 184–189.
Wu, X.Z., Zhang, T.T., Guo, J.G., Liu, Z., Yang, F.H., and Gao, X.H. 2015. Copper bioavailability, blood parameters, and nutrient balance in mink. J. Anim. Sci. 93, 176–184.
Xia, M.S., Hu, C.H., and Xu, Z.R. 2005. Effects of copper bearing montmorillonite on the growth performance, intestinal microflora and morphology of weanling pigs. Anim. Feed. Sci. Technol. 118, 307–317.
Xue, J., Li, H., Deng, X., Ma, Z., Fu, Q., and Ma, S. 2015. L-Menthone confers antidepressant-like effects in an unpredictable chronic mild stress mouse model via NLRP3 inflammasome-mediated inflammatory cytokines and central neurotransmitters. Pharmacol. Biochem. Behav. 134, 42–48.
Yang, T.H., Yuan, T.H., Hwang, Y.H., Lian, I.B., Meng, M., and Su, C.C. 2015. Increased inflammation in rheumatoid arthritis patients living where farm soils contain high levels of copper. J. Formos. Med. Assoc. 15, 1–6.
Yu, S.G., Vandenberg, G.J., and Beynen, A.C. 1995. Copper-metabolism in analbuminemic rats fed a high-copper diet. Comp. Biochem. Phys. A 110, 259–266.
Zhou, W., Kornegay, E.T., van Laar, H., Swinkels, J.W., Wong, E.A., and Lindemann, M.D. 1994. The role of feed consumption and feed efficiency in copper-stimulated growth. J. Anim. Sci. 72, 2385–2394.
Supplemental material for this article may be found at http://www.springerlink.com/content/120956.
Electronic supplementary material
About this article
Cite this article
Zhang, F., Zheng, W., Guo, R. et al. Effect of dietary copper level on the gut microbiota and its correlation with serum inflammatory cytokines in Sprague-Dawley rats. J Microbiol. 55, 694–702 (2017). https://doi.org/10.1007/s12275-017-6627-9
- inflammatory cytokines
- gut microbiota
- 16S rRNA pyrosequencing