Abstract
In China, antimicrobials and copper are used extensively as growth-promoting agents for piglets. This study aimed to characterize the role of in-feed copper in the emergence of copper-tolerant and antibiotic-resistant Enterococcus and Lactobacillus isolates in Chinese pig farms. Feces of the same eight piglets from four litters at 7 and 55 days old and their mothers were traced in order to isolate Enterococcus spp. and Lactobacillus spp.. The minimum inhibitory concentrations of 10 antimicrobials and copper sulfate were determined using an agar dilution method. The feed levels of Cu2+ for lactating sows, suckling piglets, and weaned piglets were 6, 177, and 18 mg/kg, respectively. All the 136 Enterococcus isolates were sensitive to vancomycin; and the resistance rates to penicillin, enrofloxacin, and high level streptomycin resistance increased significantly after weaning. For the 155 Lactobacillus isolates, the resistance rates to ampicillin, chloramphenicol, tetracycline, and enrofloxacin were significantly higher in weaned piglets. The ratios of copper tolerant Enterococcus and Lactobacillus isolates both increased significantly after weaning (P < 0.05). A phenotypic correlation was observed after classifying the isolates into two groups (CuSO4 MIC50 < 16 or ≧16 for enterococci; CuSO4 MIC50 < 12 or ≧12 for lactobacilli) and comparing the antimicrobial-resistant percentage of two groups. On species level, a significant increase of E. faecalis to enrofloxacin was observed in line with the increase of copper MIC (P < 0.05). The findings revealed the changes of the antibiotic resistance and copper tolerance level of enterococci and lactobacilli between suckling and weaned piglets and demonstrated that there might be a strong association between in-feed copper and increased antibiotic resistance in enterococci and lactobacilli in Chinese intensive swine farms.
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Amachawadi, R.G., Shelton, N.W., Shi, X., Vinasco, J., Dritz, S.S., Tokach, M.D., Nelssen, J.L., Scott, H.M., and Nagaraja, T.G. 2011. Selection of fecal enterococci exhibiting tcrB-mediated copper resistance in pigs fed diets supplemented with copper. Appl. Environ. Microbiol. 77, 5597–5603.
Bereket, W., Hemalatha, K., Getenet, B., Wondwossen, T., Solomon, A., Zeynudin, A., and Kannan, S. 2012. Update on bacterial nosocomial infections. Eur. Rev. Med. Pharmacol. Sci. 16, 1039–1044.
Bernardeau, M., Vernoux, J.P., Henri-Dubernet, S., and Guéguen, M. 2008. Safety assessment of dairy microorganisms: The Lactobacillus genus. Int. J. Food Microbiol. 126, 278–285.
Brooks, P.H. 2003. “Liquid feeding as a means to promote pig health”, in: Proceedings of the 3rd London swine conference: maintaining your competitive edge.(Eds J Murphy, C de Lange) pp. 83–103.
Burgos, M.J.G., López, R.L., Abriouel, H., Omar, N.B., and Galvez, A. 2009. Multilocus sequence typing of Enterococcus faecalis from vegetable foods reveals two new sequence types. Foodborne Pathog. Dis. 6, 321–327.
Butaye, P., Devriese, L., and Haesebrouck, F. 2001. Differences in antibiotic resistance patterns of Enterococcus faecalis and Enterococcus faecium strains isolated from farm and pet animals. Antimicrob. Agents Chemother. 45, 1374–1378.
CLSI. 2012. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically: Approved Standard-Ninth Edition, CLSI documemnt M07-A9, Wayne, PA: Clinical and Laboratory Standards Institute.
CLSI. 2008. Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals: Approved Standard-Third Edition. CLSI document M31-A3, Wayne, PA: Clinical and Laboratory Standards Institute.
CLSI. 2013. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-third Informational Supplement. CLSI document M100-S23, Wayne, PA: Clinical and Laboratory Standards Institute.
Danielsen, M. and Wind, A. 2003. Susceptibility of Lactobacillus spp. to antimicrobial agents. Int. J. Food Microbiol. 82, 1–11.
Dušková, M. and Karpíšková, R. 2013. Antimicrobial resistance of Lactobacilli isolated from food. Czech. J. Food Sci. 31, 27–32.
Hasman, H., Kempf, I., Chidaine, B., Cariolet, R., Ersboll, A.K., Houe, H., Bruun Hansen, H.C., and Aarestrup, F.M. 2006. Copper resistance in Enterococcus faecium, mediated by the tcrB gene, is selected by supplementation of pig feed with copper sulfate. Appl. Environ. Microbiol. 72, 5784–5789.
Hummel, A.S., Hertel, C., Holzapfel, W.H., and Franz, C.M. 2007. Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. Appl. Environ. Microbiol. 73, 730–739.
ISO (International Organization for Standardization). 2010. ISO 10932/IDF 233 standard “Milk and milk products-Determination of the minimal inhibitory concentration (MIC) of antibiotics applicable to bifidobacteria and non-enterococcal lactic acid bacteria (LAB)”.
Jin, W., Cheng, Y.F., Mao, S.Y., and Zhu, W.Y. 2014. Discovery of a novel rumen methanogen in the anaerobic fungal culture and its distribution in the rumen as revealed by real-time PCR. BMC Microbiol. 14, 104.
Kang, T.M., and Park, J.H. 2012. Isolation of Enterococcus from powdered infant and follow-on formulas, and their antibiotic susceptibilites. Food Sci. Biotechnol. 21, 1113–1118.
Li, P., Wu, D., Liu, K., Suolang, S., He, T., Liu, X., Wu, C., Wang, Y., and Lin, D. 2014. Investigation of antimicrobial resistance in Escherichia coli and enterococci isolated from Tibetan pigs. PLoS One 9, e95623.
Liu, Y., Liu, K., Lai, J., Wu, C., Shen, J., and Wang, Y. 2013. Prevalence and antimicrobial resistance of Enterococcus species of food animal origin from Beijing and Shandong Province, China. J. Appl. Microbiol. 114, 555–563.
Missotten, J., Goris, J., Michiels, J., van Coillie, E., Herman, L., De Smet, S., Dierick, N., and Heyndrickx, M. 2009. Screening of isolated lactic acid bacteria as potential beneficial strains for fermented liquid pig feed production. Anim. Feed Sci. Tech. 150, 122–138.
Mourão, J., Rae, J., Silveira, E., Freitas, A.R., Coque, T.M., Peixe, L., Antunes, P., and Novais, C. 2015. Relevance of tcrYAZB operon acquisition for Enterococcus survival at high copper concentrations under anaerobic conditions. J. Antimicrob. Chemother. 71, 560–563.
NRC. 2012. Nutrient Requirements of Swine. National Research Council, 11 ed. National Academic Press, Washington, DC, USA.
Novais, C., Freitas, A.R., Silveira, E., Antunes, P., Silva, R., Coque, T.M., and Peixe, L. 2013. Spread of multidrug-resistant Enterococcus to animals and humans: an underestimated role for the pig farm environment. J. Antimicrob. Chemother. 68, 2746–2754.
Ouoba, L.I., Lei,V., and Jensen, L.B. 2008. Resistance of potential probiotic lactic acid bacteria and bifidobacteria of African and European origin to antimicrobials: determination and transferability of the resistance genes to other bacteria. Int. J. Food Microbiol. 121, 217–224.
Panel, E.F. 2012. Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J. 10, 2740–2749.
Parvez, S., Malik, K., Ah Kang, S., and Kim, H.Y. 2006. Probiotics and their fermented food products are beneficial for health. J. Appl. Microbiol. 100, 1171–1185.
Rodriguez, L.M. and Alatossava, T. 2008. Effects of copper supplement on growth and viability of strains used as starters and adjunct cultures for Emmental cheese manufacture. J. Appl. Microbiol. 105, 1098–1106.
Schwarz, S., Silley, P., Simjee, S., Woodford, N., van Duijkeren, E., Johnson, A.P., and Gaastra, W. 2010. Editorial: assessing the antimicrobial susceptibility of bacteria obtained from animals. J. Antimicrob. Chemother. 65, 601–604.
Silveira, E., Freitas, A.R., Antunes, P., Barros, M., Campos, J., Coque, TM., Peixe, L., and Novais, C. 2014. Co-transfer of resistance to high concentrations of copper and first-line antibiotics among Enterococcus from different origins (humans, animals, the environment and foods) and clonal lineages. J. Antimicrob. Chemother. 69, 899–906.
Zhu, Y.G., Johnson, T.A., Su, J.Q., Qiao, M., Guo, G.X., Stedtfeld, R.D., Hashsham, S.A., and Tiedje, J.M. 2013. Diverse and abundant antibiotic resistance genes in Chinese swine farms. Proc. Natl. Acad. Sci. USA 110, 3435–3440.
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Zou, X., Weng, M., Ji, X. et al. Comparison of antibiotic resistance and copper tolerance of Enterococcus spp. and Lactobacillus spp. isolated from piglets before and after weaning. J Microbiol. 55, 703–710 (2017). https://doi.org/10.1007/s12275-017-6241-x
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DOI: https://doi.org/10.1007/s12275-017-6241-x