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

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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

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • 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

    Google Scholar 

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

    Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

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

    Article  Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • 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

    Article  Google Scholar 

  • 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

    CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • 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

    CAS  Google Scholar 

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Acknowledgments

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

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Authors and Affiliations

  1. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China

    Huizhi Li, Qingpo Chu, Feilong Xu, Lingling Fu, Tingting Liang, Yuan Li & Bo Zhou

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  1. Huizhi Li
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  2. Qingpo Chu
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  3. Feilong Xu
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  4. Lingling Fu
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  5. Tingting Liang
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  6. Yuan Li
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Correspondence to Bo Zhou.

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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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