Journal of Microbiology

, Volume 50, Issue 5, pp 807–812 | Cite as

In vitro development and transfer of resistance to chlortetracycline in Bacillus subtilis

  • Menghong Dai
  • Junjie Lu
  • Yulian Wang
  • Zhenli Liu
  • Zonghui Yuan


The present criteria and rules controlling the approval of the use of probiotics are limited to antibiotic resistance patterns and the presence of antibiotic resistance genes in bacteria. There is little information available in the literature regarding the risk of the usage of probiotics in the presence of antibiotic pressure. In this study we investigated the development and transfer of antibiotic resistance in Bacillus subtilis selected in vitro by chlortetracycline in a stepwise manner. Bacillus subtilis was exposed to increasing concentrations of chlortetracyclineto induce in vitro resistance to chlortetracycline, and the minimal inhibitory concentrations were determinedfor the mutants. Resistant B. subtilis were conjugated with Escherichia coli NK5449 and Enterococcus faecalis JH2-2 using the filter mating. Three B. subtilis tetracycline resistant mutants (namely, BS-1, BS-2, and BS-3) were derived in vitro. A tetracycline resistant gene, tet (K), was found in the plasmids of BS-1 and BS-2. Three conjugates (BS-1N, BS-2N, and BS-3N) were obtained when the resistant B. subtilis was conjugated with E. coli NK5449. The conjugation frequencies for the BS-1N, BS-2N, and BS-3N conjugates were 4.57×10−7, 1.4×10−7, and 1.3×10−8, respectively. The tet(K) gene was found only in the plasmids of BS-1N. These results indicate that long-term use of probiotics under antibiotic selection pressure could cause antibiotic resistance, and the resistance gene could be transferred to other bacteria. The risk arising from the use of probiotics under antibiotic pressure should be considered in the criteria and rules for the safety assessment of probiotics.


Bacillus subtilis chlortetracycline antibiotic resistance conjugation transfer tet(K) minimal inhibitory concentrations 


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Copyright information

© The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Menghong Dai
    • 1
  • Junjie Lu
    • 1
  • Yulian Wang
    • 1
  • Zhenli Liu
    • 1
  • Zonghui Yuan
    • 1
  1. 1.MOA Key Laboratory of Food Safety Evaluation / National Reference Laboratory of Veterinary Drug Residue (HZAU)Huazhong Agricultural UniversityWuhan, HubeiP. R. China

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