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Combination of antibiotics suppressed the increase of a part of ARGs in fecal microorganism of weaned pigs

Abstract

The presence of antibiotic resistance genes (ARGs) is one of the most important public health concerns. Six tetracycline resistance genes (TRGs—tetA, tetC, tetL, tetO, tetW, and tetX) were quantified using real-time quantitative polymerase chain reaction (qPCR) in the fecal microorganisms of weaned pigs. Two hundred 35-day-old weaned pigs were fed different dietary antibiotics for 28 days: (1) no antibiotic as the control treatment (CT); (2) chlortetracycline, bacitracin zinc and colistin sulfate (CBC); (3) bacitracin zinc and colistin sulfate (BC); and (4) chlortetracycline (CTC). The detection frequencies (DFs) of tetC, tetL, and tetW were 100 %; and the DFs of tetA, tetD, tetM, tetO, and tetX were 65 %. The relative abundances (tet/16S rRNA gene copy numbers) of six tet genes (tetA, tetC, tetL, tetO, tetW and tetX) were between 1.5 × 10−4 and 2.0 × 10−1. In the group CTC, the relative abundances of tetC (P < 0.01), tetL (P < 0.01), tetO (P < 0.05), tetW (P < 0.01), and tetX (P < 0.01) were greater than those of the group CT. Compared with the group CTC, the relative abundances of tetC (P < 0.01), tetL (P < 0.01), and tetW (P < 0.01) were decreased in the CBC and BC groups. These results indicate that a combination of different antibiotics suppressed the abundance increase of a part of tet genes, which suggests that a combination of antibiotics produces multiple selection pressures on fecal microorganism of pigs.

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Abbreviations

CTC:

Chlortetracycline

ARGs:

Antibiotic resistance genes

TRGs:

Tetracycline resistance genes

Tet :

Tetracycline

qPCR:

Real-time quantitative polymerase chain reaction

DFs:

Detection frequencies

References

  1. Agga GE, Scott HM, Amachawadi RG, Nagaraja TG, Vinasco J, Bai J, Norby B, Renter DG, Dritz SS, Nelssen JL, Tokach MD (2014) Effects of chlortetracycline and copper supplementation on antimicrobial resistance of fecal Escherichia coli from weaned pigs. Prev Vet Med 114:231–246

  2. Aminov RI, Garrigues-Jeanjean N, Mackie RI (2001) Molecular ecology of tetracycline resistance: development and validation of primers for detection of tetracycline resistance genes encoding ribosomal protection proteins. Appl Environ Microbiol 67:22–32

  3. Barkovskii AL, Bridges C (2012) Persistence and profiles of tetracycline resistance genes in swine farms and impact of operational practices on their occurrence in farms’ vicinities. Water Air Soil Pollut 223:49–62

  4. Bennett PM (2008) Plasmid encoded antibiotic resistance: acquisition and transfer of antibiotic resistance genes in bacteria. Br J Pharmacol 153:S347–S357

  5. Bineva I, Karaivanov L (1983) In vitro action of zinc bacitracin to eliminate drug resistance in Escherichia coli transconjugates. Vet Med Nauki 21:36–42

  6. Broadway P, Carroll J, Callaway T (2014) Antibiotic use in livestock production. Agric Food Anal Bacteriol 4:76–85

  7. Bush K, Courvalin P, Dantas G, Davies J, Eisenstein B, Huovinen P, Jacoby GA, Kishony R, Kreiswirth BN, Kutter E (2011) Tackling antibiotic resistance. Nat Rev Microbiol 9:894–896

  8. Chen C, Li J, Chen P, Ding R, Zhang P, Li X (2014) Occurrence of antibiotics and antibiotic resistances in soils from wastewater irrigation areas in Beijing and Tianjin, China. Environ Pollut 193:94–101

  9. Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev 65:232–260

  10. Coffman JR (Chairman) National Research Council (1999) The use of drugs in food animals: benefits and risks. National Academy Press, Washington, DC, p 28

  11. Cromwell GL (2002) Why and how antibiotics are used in swine production. Anim Biotechnol 13:7–27

  12. D’Costa VM, King CE, Kalan L, Morar M, Sung WW, Schwarz C, Froese D, Zazula G, Calmels F, Debruyne R (2011) Antibiotic resistance is ancient. Nature 477:457–461

  13. Dudhagara PR (2014) Phenotypic characterization and antibiotics combination approach to control the methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from the hospital derived fomites. Asian J Med Sci 2:72–78

  14. Fan W, Hamilton T, Webster-Sesay S, Nikolich MP, Lindler LE (2007) Multiplex real-time SYBR green I PCR assay for detection of tetracycline efflux genes of Gram-negative bacteria. Mol Cell Probes 21:245–256

  15. Ghosh S, Ramsden SJ, LaPara TM (2009) The role of anaerobic digestion in controlling the release of tetracycline resistance genes and class 1 integrons from municipal wastewater treatment plants. Appl Microbiol Biotechnol 84:791–796

  16. Gong J, Yu H, Liu T, Gill J, Chambers J, Wheatcroft R, Sabour P (2008) Effects of zinc bacitracin, bird age and access to range on bacterial microbiota in the ileum and caeca of broiler chickens. J Appl Microbiol 104:1372–1382

  17. Gordon RC, Barrett FF, Clark DJ (1972) Influence of several antibiotics, singly and in combination, on the growth of Listeria monocytogenes. J Pediatr 80:667–670

  18. Guyonnet J, Manco B, Baduel L, Kaltsatos V, Aliabadi M, Lees P (2010) Determination of a dosage regimen of colistin by pharmacokinetic/pharmacodynamic integration and modeling for treatment of GIT disease in pigs. Res Vet Sci 88:307–314

  19. Harvey R, Funk J, Wittum TE, Hoet AE (2009) A metagenomic approach for determining prevalence of tetracycline resistance genes in the fecal flora of conventionally raised feedlot steers and feedlot steers raised without antimicrobials. Am J Vet Res 70:198–202

  20. Holman DB, Chenier MR (2013) Impact of subtherapeutic administration of tylosin and chlortetracycline on antimicrobial resistance in farrow-to-finish swine. FEMS Microbiol Ecol 85:1–13

  21. Huang X, Liu C, Li K, Su J, Zhu G, Liu L (2015) Performance of vertical up-flow constructed wetlands on swine wastewater containing tetracyclines and tet genes. Water Res 70:109–117

  22. Jindal A, Kocherginskaya S, Mehboob A, Robert M, Mackie RI, Raskin L, Zilles JL (2006) Antimicrobial use and resistance in swine waste treatment systems. Appl Environ Microbiol 72:7813–7820

  23. Joy SR, Li X, Snow DD, Gilley JE, Woodbury B, Bartelt-Hunt SL (2014) Fate of antimicrobials and antimicrobial resistance genes in simulated swine manure storage. Sci Total Environ 481:69–74

  24. Kay P, Blackwell PA, Boxall AB (2005) Column studies to investigate the fate of veterinary antibiotics in clay soils following slurry application to agricultural land. Chemosphere 60:497–507

  25. Laht M, Karkman A, Voolaid V, Ritz C, Tenson T, Virta M, Kisand V (2014) Abundances of tetracycline, sulphonamide and beta-lactam antibiotic resistance genes in conventional wastewater treatment plants (WWTPs) with different waste load. PLoS One 9:e103705

  26. Liu M, Zhang Y, Yang M, Tian Z, Ren L, Zhang S (2012) Abundance and distribution of tetracycline resistance genes and mobile elements in an oxytetracycline production wastewater treatment system. Environ Sci Technol 46:7551–7557

  27. Looft T, Johnson TA, Allen HK, Bayles DO, Alt DP, Stedtfeld RD, Sul WJ, Stedtfeld TM, Chai B, Cole JR, Hashsham SA, Tiedje JM, Stanton TB (2012) In-feed antibiotic effects on the swine intestinal microbiome. Proc Natl Acad Sci U S A 109:1691–1696

  28. Luo Y, Xu L, Rysz M, Wang Y, Zhang H, Alvarez PJ (2011) Occurrence and transport of tetracycline, sulfonamide, quinolone, and macrolide antibiotics in the Haihe River basin, China. Environ Sci Technol 45:1827–1833

  29. Marshall BM, Levy SB (2011) Food animals and antimicrobials: impacts on human health. Clin Microbiol Rev 24:718–733

  30. Mu Q, Li J, Sun Y, Mao D, Wang Q, Luo Y (2015) Occurrence of sulfonamide-, tetracycline-, plasmid-mediated quinolone- and macrolide-resistance genes in livestock feedlots in northern China. Environ Sci Pollut Res Int 22:6932–6940

  31. Ng LK, Martin I, Alfa M, Mulvey M (2001) Multiplex PCR for the detection of tetracycline resistant genes. Mol Cell Probes 15:209–215

  32. Phillips I, Casewell M, Cox T, De Groot B, Friis C, Jones R, Nightingale C, Preston R, Waddell J (2004) Does the use of antibiotics in food animals pose a risk to human health? A critical review of published data. J Antimicrob Chemother 53:28–52

  33. Pindell M, Cull K, Doran K, Dickison H (1959) Absorption and excretion studies on tetracycline. J Pharmacol Exp Ther 125:287–294

  34. Roberts MC (2005) Update on acquired tetracycline resistance genes. FEMS Microbiol Lett 245:195–203

  35. Roberts MC (2011) Environmental macrolide-lincosamide-streptogramin and tetracycline resistant bacteria. Front Microbiol 2:40

  36. Sarmah AK, Meyer MT, Boxall ABA (2006) A global perspective on the use, sales, exposure pathways, occurrence, fate and effects of veterinary antibiotics (VAs) in the environment. Chemosphere 65:725–759

  37. Simpson JM, McCracken VJ, White BA, Gaskins HR, Mackie RI (1999) Application of denaturant gradient gel electrophoresis for the analysis of the porcine gastrointestinal microbiota. J Microbiol Methods 36:167–179

  38. Smith DL, Harris AD, Johnson JA, Silbergeld EK, Morris JG Jr (2002) Animal antibiotic use has an early but important impact on the emergence of antibiotic resistance in human commensal bacteria. Proc Natl Acad Sci U S A 99:6434–6439

  39. Speer BS, Shoemaker NB, Salyers AA (1992) Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance. Clin Microbiol Rev 5:387–399

  40. Szczepanowski R, Linke B, Krahn I, Gartemann KH, Gutzkow T, Eichler W, Puhler A, Schluter A (2009) Detection of 140 clinically relevant antibiotic-resistance genes in the plasmid metagenome of wastewater treatment plant bacteria showing reduced susceptibility to selected antibiotics. Microbiology 155:2306–2319

  41. Tsai YL, Olson BH (1991) Rapid method for direct extraction of DNA from soil and sediments. Appl Environ Microbiol 57:1070–1074

  42. Udikovic-Kolic N, Wichmann F, Broderick NA, Handelsman J (2014) Bloom of resident antibiotic-resistant bacteria in soil following manure fertilization. Proc Natl Acad Sci U S A 111:15202–15207

  43. Walsh C (2003) Where will new antibiotics come from? Nat Rev Microbiology 1:65–70

  44. Wang HF, Zhu WY, Yao W, Liu JX (2007) DGGE and 16S rDNA sequencing analysis of bacterial communities in colon content and feces of pigs fed whole crop rice. Anaerobe 13:127–133

  45. Watine J, Bourrel C, Dubourdieu B, Gineston J, Bories P, Durand M, Marre A, Formosa F, Brunel M, Palliez J (1994) Susceptibility of multiresistant serotype 012 Pseudomonas Aeruginosa to fosfomycin in combination with other antibiotics. Pathol Biol 42:293–295

  46. Yoon SY, Eo SK, Kim YS, Lee CK, Han SS (1994) Antimicrobial activity of Ganoderma lucidum extract alone and in combination with some antibiotics. Arch Pharm Res 17:438–442

  47. Zhu YG, Johnson TA, Su JQ, Qiao M, Guo GX, Stedtfeld RD, Hashsham SA, Tiedje JM (2013) Diverse and abundant antibiotic resistance genes in Chinese swine farms. Proc Natl Acad Sci U S A 110:3435–3440

  48. Zoetendal EG, Akkermans AD, De Vos WM (1998) Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria. Appl Environ Microbiol 64:3854–3859

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Acknowledgments

We are grateful to Jiangsu Yong-kang Agricultural and Animal Husbandry Co., Ltd. for the experimental animals.

Author information

Correspondence to Bo Zhou.

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All the authors agreed with this submission.

Conflict of interest

The authors declare that they have no conflicts of interest.

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Responsible editor: Robert Duran

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Li, H., Chu, Q., Xu, F. et al. Combination of antibiotics suppressed the increase of a part of ARGs in fecal microorganism of weaned pigs. Environ Sci Pollut Res 23, 18183–18191 (2016). https://doi.org/10.1007/s11356-016-7004-7

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Keywords

  • Chlortetracycline
  • Fecal microorganism
  • Pigs
  • tet genes
  • Tetracycline resistance
  • qPCR