Antibiotics as a selective driver for conjugation dynamics

It is generally assumed that antibiotics can promote horizontal gene transfer. However, because of a variety of confounding factors that complicate the interpretation of previous studies, the mechanisms by which antibiotics modulate horizontal gene transfer remain poorly understood. In particular, it is unclear whether antibiotics directly regulate the efficiency of horizontal gene transfer, serve as a selection force to modulate population dynamics after such gene transfer has occurred, or both. Here, we address this question by quantifying conjugation dynamics in the presence and absence of antibiotic-mediated selection. Surprisingly, we find that sublethal concentrations of antibiotics from the most widely used classes do not significantly increase the conjugation efficiency. Instead, our modelling and experimental results demonstrate that conjugation dynamics are dictated by antibiotic-mediated selection, which can both promote and suppress conjugation dynamics. Our findings suggest that the contribution of antibiotics to the promotion of horizontal gene transfer may have been overestimated. These findings have implications for designing effective antibiotic treatment protocols and for assessing the risks of antibiotic use.

[1]  W. Hoekstra,et al.  On the role of the recipient cell during conjugation in Escherichia coli , 2004, Antonie van Leeuwenhoek.

[2]  Wolf-Dietrich Hardt,et al.  Gut inflammation can boost horizontal gene transfer between pathogenic and commensal Enterobacteriaceae , 2012, Proceedings of the National Academy of Sciences.

[3]  A. Martner,et al.  Transfer of an ampicillin resistance gene between two Escherichia coli strains in the bowel microbiota of an infant treated with antibiotics. , 2007, The Journal of antimicrobial chemotherapy.

[4]  Yue-zhong Li,et al.  Combined treatment with the antibiotics kanamycin and streptomycin promotes the conjugation of Escherichia coli. , 2013, FEMS microbiology letters.

[5]  Barth F. Smets,et al.  Horizontal gene transfer: perspectives at a crossroads of scientific disciplines , 2005, Nature Reviews Microbiology.

[6]  A. Stevens,et al.  Tetracycline regulation of genes on Bacteroides conjugative transposons , 1993, Journal of bacteriology.

[7]  J. Dewulf,et al.  In situ ESBL conjugation from avian to human Escherichia coli during cefotaxime administration , 2011, Journal of applied microbiology.

[8]  R. Aminov Horizontal Gene Exchange in Environmental Microbiota , 2011, Front. Microbio..

[9]  G. Whitesides,et al.  Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane). , 1998, Analytical chemistry.

[10]  J. M. Schuurmans,et al.  Effect of growth rate and selection pressure on rates of transfer of an antibiotic resistance plasmid between E. coli strains. , 2014, Plasmid.

[11]  L. Frost,et al.  The role of the pilus in recipient cell recognition during bacterial conjugation mediated by F‐like plasmids , 1994, Molecular microbiology.

[12]  Chase L. Beisel,et al.  Repurposing endogenous type I CRISPR-Cas systems for programmable gene repression , 2014, Nucleic acids research.

[13]  J. Davison,et al.  Genetic exchange between bacteria in the environment. , 1999, Plasmid.

[14]  D. Nair,et al.  Extended-spectrum β-lactamases in Gram Negative Bacteria , 2010, Journal of global infectious diseases.

[15]  F. Aarestrup,et al.  Selection and Persistence of CTX-M-Producing Escherichia coli in the Intestinal Flora of Pigs Treated with Amoxicillin, Ceftiofur, or Cefquinome , 2008, Antimicrobial Agents and Chemotherapy.

[16]  Eran Segal,et al.  Growth dynamics of gut microbiota in health and disease inferred from single metagenomic samples , 2015, Science.

[17]  N. Kroer,et al.  Effects of stress and other environmental factors on horizontal plasmid transfer assessed by direct quantification of discrete transfer events. , 2007, FEMS microbiology ecology.

[18]  Miriam Barlow,et al.  What antimicrobial resistance has taught us about horizontal gene transfer. , 2009, Methods in molecular biology.

[19]  Y. Carmeli,et al.  Transfer of Carbapenem-Resistant Plasmid from Klebsiella pneumoniae ST258 to Escherichia coli in Patient , 2010, Emerging infectious diseases.

[20]  R. Curtiss,et al.  Energy requirements for specific pair formation during conjugation in Escherichia coli K-12 , 1967, Journal of bacteriology.

[21]  F. M. Stewart,et al.  The kinetics of conjugative plasmid transmission: fit of a simple mass action model. , 1979, Plasmid.

[22]  Diarmaid Hughes,et al.  Microbiological effects of sublethal levels of antibiotics , 2014, Nature Reviews Microbiology.

[23]  J. Blázquez,et al.  Antimicrobials as promoters of genetic variation. , 2012, Current opinion in microbiology.

[24]  Non-invasive determination of conjugative transfer of plasmids bearing antibiotic-resistance genes in biofilm-bound bacteria: effects of substrate loading and antibiotic selection , 2012, Applied Microbiology and Biotechnology.

[25]  M. Millar,et al.  Beta-lactam antibiotics increase the frequency of plasmid transfer in Staphylococcus aureus. , 1986, The Journal of antimicrobial chemotherapy.

[26]  S Falkow,et al.  FACS-optimized mutants of the green fluorescent protein (GFP). , 1996, Gene.

[27]  E. Egelman,et al.  The structure of F-pili. , 2009, Journal of molecular biology.

[28]  M Zatyka,et al.  Control of genes for conjugative transfer of plasmids and other mobile elements. , 1998, FEMS microbiology reviews.

[29]  U. Alon,et al.  A comprehensive library of fluorescent transcriptional reporters for Escherichia coli , 2006, Nature Methods.

[30]  K. Cheah,et al.  The F plasmid carries an IS3 insertion within finO. , 1986, Journal of general microbiology.

[31]  M. Maiden,et al.  Horizontal genetic exchange, evolution, and spread of antibiotic resistance in bacteria. , 1998, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[32]  M. Iacaruso,et al.  New Algorithm to Determine True Colocalization in Combination with Image Restoration and Time-Lapse Confocal Microscopy to Map Kinases in Mitochondria , 2011, PloS one.

[33]  A. Salyers,et al.  In the driver's seat: the Bacteroides conjugative transposons and the elements they mobilize , 1995, Journal of bacteriology.

[34]  D. Mazel,et al.  Antibiotic resistance in microbes , 1999, Cellular and Molecular Life Sciences CMLS.

[35]  A. D. Russell,et al.  Effect of some antibiotics and biocides on plasmid transfer in Staphylococcus aureus. , 1991, The Journal of applied bacteriology.

[36]  François Taddei,et al.  Genetic information transfer promotes cooperation in bacteria , 2014, Proceedings of the National Academy of Sciences.

[37]  Stefan Wuertz,et al.  Studying plasmid horizontal transfer in situ: a critical review , 2005, Nature Reviews Microbiology.

[38]  Lucinda E. Maddera,et al.  F-pili dynamics by live-cell imaging , 2008, Proceedings of the National Academy of Sciences.

[39]  Improving conjugation efficacy of Sorangium cellulosum by the addition of dual selection antibiotics , 2008, Journal of Industrial Microbiology & Biotechnology.

[40]  A. Salyers,et al.  Characterization of Genes Involved in Modulation of Conjugal Transfer of the Bacteroides Conjugative Transposon CTnDOT , 2002, Journal of bacteriology.

[41]  A. Salyers,et al.  Tetracycline-dependent appearance of plasmidlike forms in Bacteroides uniformis 0061 mediated by conjugal Bacteroides tetracycline resistance elements , 1988, Journal of bacteriology.

[42]  L. Frost,et al.  F- phenocopies: characterization of expression of the F transfer region in stationary phase. , 1998, Microbiology.

[43]  H. Bujard,et al.  Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. , 1997, Nucleic acids research.

[44]  John W. Beaber,et al.  SOS response promotes horizontal dissemination of antibiotic resistance genes , 2004, Nature.

[45]  Lingchong You,et al.  Spatiotemporal modulation of biodiversity in a synthetic chemical-mediated ecosystem , 2009, Nature chemical biology.

[46]  Søren J. Sørensen,et al.  Conjugative plasmids: vessels of the communal gene pool , 2009, Philosophical Transactions of the Royal Society B: Biological Sciences.

[47]  Christopher M Thomas,et al.  Mechanisms of, and Barriers to, Horizontal Gene Transfer between Bacteria , 2005, Nature Reviews Microbiology.

[48]  D. Newby,et al.  The role of selective pressure and selfish DNA in horizontal gene transfer and soil microbial community adaptation , 2002 .

[49]  M. Jonker,et al.  Factors That Affect Transfer of the IncI1 β-Lactam Resistance Plasmid pESBL-283 between E. coli Strains , 2015, PloS one.