Quantitative and qualitative impact of hospital effluent on dissemination of the integron pool

There is increasing evidence that human activity, and especially the resulting effluent, has a major role in the dissemination of bacterial antibiotic-resistance determinants in the environment. Hospitals are the major antibiotic consumers and thus facilitate the spread of antibiotic resistance. Questions are increasingly being raised about the management of hospital effluents, but their involvement in antibiotic-resistance dissemination has never been assessed. Integrons are a paradigm of genetic transfer between the environmental resistome and both commensal and pathogenic bacteria. In order to assess the impact of hospital activities on antibiotic-resistance dissemination in the environment, we monitored integrons and their gene cassettes in hospital effluents, and their release in the environment. We found that bacterial communities present in a hospital effluent contained a high proportion of integrons. In terms of both their gene cassette diversity and gene cassette arrays, the urban effluent and municipal wastewater treatment plant (WWTP) influent were most similar, whereas the hospital effluent and recirculation sludge exhibited very specific patterns. We found that anthropogenic activities led to the release of abundant integrons and antibiotic-resistance gene cassettes, but we observed no specific impact of hospital activities on the receiving environment. Furthermore, although the WWTP did not reduce the normalized integron copy number, it reduced the diversity of gene cassette arrays contained in the raw wastewater, underlining the effect of the biological treatment on the anthropogenic integron pool arriving at the WWTP.

[1]  R. Hall,et al.  A novel family of potentially mobile DNA elements encoding site‐specific gene‐integration functions: integrons , 1989, Molecular microbiology.

[2]  Chantal D. Larose,et al.  PCR mapping of integrons reveals several novel combinations of resistance genes , 1995, Antimicrobial agents and chemotherapy.

[3]  K. Schleifer,et al.  Phylogenetic identification and in situ detection of individual microbial cells without cultivation. , 1995, Microbiological reviews.

[4]  K Kümmerer,et al.  Drugs in the environment: emission of drugs, diagnostic aids and disinfectants into wastewater by hospitals in relation to other sources--a review. , 2001, Chemosphere.

[5]  P. Nordmann,et al.  GES-2, a Class A β-Lactamase fromPseudomonas aeruginosa with Increased Hydrolysis of Imipenem , 2001, Antimicrobial Agents and Chemotherapy.

[6]  James R. Cole,et al.  rrndb: the Ribosomal RNA Operon Copy Number Database , 2001, Nucleic Acids Res..

[7]  D. Mazel,et al.  Bacterial resistance evolution by recruitment of super‐integron gene cassettes , 2002, Molecular microbiology.

[8]  P. Nordmann,et al.  aeruginosa Pseudomonas Class 1 Integron In 120 in-Lactamase , and the β Extended-Spectrum BEL-1 , a Novel Clavulanic Acid-Inhibited , 2005 .

[9]  E. Wellington,et al.  Incidence of Class 1 Integrons in a Quaternary Ammonium Compound-Polluted Environment , 2005, Antimicrobial Agents and Chemotherapy.

[10]  Didier Mazel,et al.  Integrons: agents of bacterial evolution , 2006, Nature Reviews Microbiology.

[11]  Célia M Manaia,et al.  Antimicrobial resistance patterns in Enterobacteriaceae isolated from an urban wastewater treatment plant. , 2007, FEMS microbiology ecology.

[12]  W. Doolittle,et al.  Integron-associated gene cassettes in Halifax Harbour: assessment of a mobile gene pool in marine sediments. , 2008, Environmental microbiology.

[13]  Yan Boucher,et al.  The Evolution of Class 1 Integrons and the Rise of Antibiotic Resistance , 2008, Journal of bacteriology.

[14]  F. Baquero,et al.  Antibiotics and antibiotic resistance in water environments. , 2008, Current opinion in biotechnology.

[15]  S. Hardwick,et al.  Quantification of class 1 integron abundance in natural environments using real-time quantitative PCR. , 2008, FEMS microbiology letters.

[16]  R. Stepanauskas,et al.  Influence of industrial contamination on mobile genetic elements: class 1 integron abundance and gene cassette structure in aquatic bacterial communities , 2008, The ISME Journal.

[17]  Tong Zhang,et al.  Characterization and quantification of class 1 integrons and associated gene cassettes in sewage treatment plants , 2009, Applied Microbiology and Biotechnology.

[18]  S. Hardwick,et al.  Gene cassettes encoding resistance to quaternary ammonium compounds: a role in the origin of clinical class 1 integrons? , 2009, The ISME Journal.

[19]  J. Martínez,et al.  Environmental pollution by antibiotics and by antibiotic resistance determinants. , 2009, Environmental pollution.

[20]  C. Médigue,et al.  Genome sequence of Vibrio splendidus: an abundant planctonic marine species with a large genotypic diversity. , 2009, Environmental microbiology.

[21]  H. Heuer,et al.  Diverse aadA gene cassettes on class 1 integrons introduced into soil via spread manure. , 2009, Research in microbiology.

[22]  Alfred Pühler,et al.  Detection of 140 clinically relevant antibiotic-resistance genes in the plasmid metagenome of wastewater treatment plant bacteria showing reduced susceptibility to selected antibiotics. , 2009, Microbiology.

[23]  E. Coiera,et al.  Gene cassettes and cassette arrays in mobile resistance integrons. , 2009, FEMS microbiology reviews.

[24]  G. Cambray,et al.  The SOS Response Controls Integron Recombination , 2009, Science.

[25]  Tong Zhang,et al.  Class 1 integronase gene and tetracycline resistance genes tetA and tetC in different water environments of Jiangsu Province, China , 2009, Ecotoxicology.

[26]  M. Baclet,et al.  Quantitative multiplex real-time PCR for detecting class 1, 2 and 3 integrons. , 2010, The Journal of antimicrobial chemotherapy.

[27]  C. Manaia,et al.  Factors influencing antibiotic resistance burden in municipal wastewater treatment plants , 2010, Applied Microbiology and Biotechnology.

[28]  P. H. Roy,et al.  Effect of attC structure on cassette excision by integron integrases , 2011, Mobile DNA.

[29]  K. Jankowska,et al.  Antimicrobial resistance of fecal indicators in municipal wastewater treatment plant. , 2010, Water research.

[30]  K. Smalla,et al.  Wastewater bacterial communities bring together broad-host range plasmids, integrons and a wide diversity of uncharacterized gene cassettes. , 2010, Research in microbiology.

[31]  H. Stokes,et al.  Class 1 integrons in benthic bacterial communities: abundance, association with Tn402-like transposition modules and evidence for coselection with heavy-metal resistance. , 2010, FEMS microbiology ecology.

[32]  Y. Kamagata,et al.  Marine integrons containing novel integrase genes, attachment sites, attI, and associated gene cassettes in polluted sediments from Suez and Tokyo Bays , 2011, The ISME Journal.

[33]  P. McNamara,et al.  Tertiary-treated municipal wastewater is a significant point source of antibiotic resistance genes into Duluth-Superior Harbor. , 2011, Environmental science & technology.

[34]  W. Doolittle,et al.  Coral-mucus-associated Vibrio integrons in the Great Barrier Reef: genomic hotspots for environmental adaptation , 2011, The ISME Journal.

[35]  A. Boxall,et al.  Impacts of anthropogenic activity on the ecology of class 1 integrons and integron-associated genes in the environment , 2011, The ISME Journal.

[36]  Michael R Gillings,et al.  Gene flow, mobile genetic elements and the recruitment of antibiotic resistance genes into Gram-negative pathogens. , 2011, FEMS microbiology reviews.

[37]  R. Norman,et al.  Characterization and Quantitation of a Novel β-Lactamase Gene Found in a Wastewater Treatment Facility and the Surrounding Coastal Ecosystem , 2011, Applied and Environmental Microbiology.

[38]  K. Nielsen,et al.  Integrons , 2012, Mobile Genetic Elements.

[39]  Magali Casellas,et al.  Integron Involvement in Environmental Spread of Antibiotic Resistance , 2012, Front. Microbio..

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

[41]  G. Dantas,et al.  The Shared Antibiotic Resistome of Soil Bacteria and Human Pathogens , 2012, Science.

[42]  A. P. Williams,et al.  The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria. , 2013, The Lancet. Infectious diseases.

[43]  M. Casellas,et al.  An antibiotic-resistant class 3 integron in an Enterobacter cloacae isolate from hospital effluent. , 2013, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[44]  Teresa M. Coque,et al.  Antibiotic resistance shaping multi-level population biology of bacteria , 2013, Front. Microbiol..

[45]  C. Manaia,et al.  Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. , 2013, The Science of the total environment.

[46]  Alexandro Rodríguez-Rojas,et al.  Antibiotics and antibiotic resistance: a bitter fight against evolution. , 2013, International journal of medical microbiology : IJMM.

[47]  M. Pons,et al.  Dynamic assessment of the floc morphology, bacterial diversity, and integron content of an activated sludge reactor processing hospital effluent. , 2013, Environmental science & technology.

[48]  T. Schwartz,et al.  Sub-inhibitory concentrations of antibiotics and wastewater influencing biofilm formation and gene expression of multi-resistant Pseudomonas aeruginosa wastewater isolates , 2013, Environmental Science and Pollution Research.