Comparison of Horizontal blaCTX-M Gene Transfer via Conjugation among Extended Spectrum β-Lactamases Producing Escherichia coli Isolates from Patients with Urinary Tract Infection, Their Animals, and Environment

Background The dissemination of the extended spectrum β-lactamases (ESBL) producing E. coli poses a significant public health problem. Understanding the efficiency and frequency of horizontal gene transfer via conjugation of ESBL producing E. coli is imperative towards devising prevention and control measures. This study compared the frequencies and efficiencies of horizontal blaCTX-M gene transfer via conjugation among Escherichia coli isolates from urine and gastrointestinal tract (GIT) of patients with urinary tract infection (UTI), their animals and environment. Methods Horizontal blaCTX-M gene transfer via conjugation by a broth mating experiment was performed using 50 confirmed ESBL producing E. coli isolates as donors and Escherichia coli J53 (F−, met, pro, Azr), as the recipient. The transconjugants were detected and their frequencies and efficiencies of conjugation were measured and compared between ESBL producing E. coli isolates multi-sourced from urine, GIT, animals and environment. Antimicrobial susceptibility testing of all resulting transconjugants was performed. DNA was extracted from all transconjugants to confirm the presence and the acquisition of blaCTX-M gene. Results Out of 50 ESBL producing E. coli isolates harboring blaCTX-M gene, 37 (74.0%) successfully exercised horizontal gene transfer through conjugation. All transconjugants were confirmed phenotypically and genotypically by PCR. Of note, all of the isolates from environment 100.0% (7/7) performed conjugation, exhibiting the highest transfer efficiency, followed by isolates from urine and animals, with the conjugation transfer efficiency of 77.8% (14/18) and 76.1% (10/13), respectively. The isolates from the environment conjugated with a significant more efficiency than those from the GIT [Two-sample test of proportions; p-value = 0.0119]. The overall conjugation transfer frequencies ranged from 0.4 × 10-14 – 5.5 × 10-11 per donor cells with the highest median conjugation transfer frequency observed among isolates from animal (3.23 × 10-12 [IQR: 0.70 × 10-12 – 7.22 × 10-12]) followed by that of isolates from the environment (1.60 × 10-12 [IQR: 0.30 × 10-12 – 5.0 × 10-12]). Conclusion ESBL producing E. coli from human, animals and environment exercises horizontal blaCTX-M gene transfer efficiently with the highest occurrence among isolates from the environment and animals. The antimicrobial resistance control and prevention strategies should be widened up to explore strategies to prevent horizontal AMR gene transfer.

[1]  S. Mshana,et al.  Conjugative Plasmids Disseminating CTX-M-15 among Human, Animals and the Environment in Mwanza Tanzania: A Need to Intensify One Health Approach , 2021, Antibiotics.

[2]  M. Matee,et al.  Occurrence of Multidrug Resistant Escherichia coli in Raw Meat and Cloaca Swabs in Poultry Processed in Slaughter Slabs in Dar es Salaam, Tanzania , 2021, Antibiotics.

[3]  A. Vitas,et al.  The Aquatic Ecosystem, a Good Environment for the Horizontal Transfer of Antimicrobial Resistance and Virulence-Associated Factors Among Extended Spectrum β-lactamases Producing E. coli , 2020, Microorganisms.

[4]  N. Hassuna,et al.  Molecular characterization of Extended-spectrum β lactamase- producing E. coli recovered from community-acquired urinary tract infections in Upper Egypt , 2020, Scientific Reports.

[5]  N. Klibi,et al.  Detection of CTX-M-15 harboring Escherichia coli isolated from wild birds in Tunisia , 2018, BMC Microbiology.

[6]  F. de la Cruz,et al.  Conjugation Inhibitors and Their Potential Use to Prevent Dissemination of Antibiotic Resistance Genes in Bacteria , 2017, Front. Microbiol..

[7]  Trinad Chakraborty,et al.  Predominance of CTX-M-15 among ESBL Producers from Environment and Fish Gut from the Shores of Lake Victoria in Mwanza, Tanzania , 2016, Front. Microbiol..

[8]  Ole Lund,et al.  A Bacterial Analysis Platform: An Integrated System for Analysing Bacterial Whole Genome Sequencing Data for Clinical Diagnostics and Surveillance , 2016, PloS one.

[9]  T. Chakraborty,et al.  Predictors of blaCTX-M-15 in varieties of Escherichia coli genotypes from humans in community settings in Mwanza, Tanzania , 2016, BMC Infectious Diseases.

[10]  Petra F. G. Wolffs,et al.  Dissemination of Antimicrobial Resistance in Microbial Ecosystems through Horizontal Gene Transfer , 2016, Front. Microbiol..

[11]  T. Chakraborty,et al.  Multiple ESBL-Producing Escherichia coli Sequence Types Carrying Quinolone and Aminoglycoside Resistance Genes Circulating in Companion and Domestic Farm Animals in Mwanza, Tanzania, Harbor Commonly Occurring Plasmids , 2016, Front. Microbiol..

[12]  E. Wellington,et al.  Waste water effluent contributes to the dissemination of CTX-M-15 in the natural environment , 2014, The Journal of antimicrobial chemotherapy.

[13]  Joseph M. Campos,et al.  Extended-spectrum Beta-lactamases: A Brief Clinical Update , 2011, The Pediatric infectious disease journal.

[14]  E. Domann,et al.  Multiple ST clonal complexes, with a predominance of ST131, of Escherichia coli harbouring blaCTX-M-15 in a tertiary hospital in Tanzania. , 2011, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[15]  P. Savelkoul,et al.  Extended-Spectrum β-Lactamase Genes of Escherichia coli in Chicken Meat and Humans, the Netherlands , 2011, Emerging infectious diseases.

[16]  Eric Bapteste,et al.  Network analyses structure genetic diversity in independent genetic worlds , 2009, Proceedings of the National Academy of Sciences.

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

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

[19]  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.

[20]  P M Bennett,et al.  Plasmid encoded antibiotic resistance: acquisition and transfer of antibiotic resistance genes in bacteria , 2008, British journal of pharmacology.

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

[22]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

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

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

[25]  H. Smith.  Transfer of antibiotic resistance from animal and human strains of Escherichia coli to resident E. coli in the alimentary tract of man. , 1969, The Veterinary record.