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Temperature and Nutrient Limitations Decrease Transfer of Conjugative IncP-1 Plasmid pKJK5 to Wild Escherichia coli

Rebeca Pallares-Vega1,2, Gonçalo Macedo1,3, Michael S M Brouwer4

  • 1Wetsus, European Centre of Excellence for Sustainable Water Technology, Leeuwarden, Netherlands.

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Summary

Antibiotic resistance genes spread via plasmids in gut bacteria. Even in harsh environmental conditions like low nutrients and cold, plasmid transfer occurs, but nutrient availability is the main limiting factor.

Keywords:
AMRantimicrobial resistanceenvironmental conditionshorizontal gene transfersoil extract agarsynthetic wastewater

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Area of Science:

  • Environmental microbiology
  • Molecular biology
  • Antimicrobial resistance

Background:

  • Antibiotic resistance genes (ARGs) in fecal Enterobacteriaceae can persist in the environment.
  • Laboratory conditions may overestimate plasmid transfer rates in natural ecosystems.

Purpose of the Study:

  • To assess the conjugative transfer of an IncP-1 plasmid (pKJK5) to three natural Escherichia coli strains under varying environmental conditions.
  • To understand the impact of temperature and nutrient limitations on plasmid dissemination.

Main Methods:

  • Filter mating experiments were conducted using three natural Escherichia coli strains harboring extended-spectrum beta-lactamases.
  • Matings were performed under optimal laboratory conditions (LB medium, 37°C) and environmentally relevant conditions (25°C, 15°C, 9°C; synthetic wastewater; soil extract).

Main Results:

  • High plasmid transfer frequencies (up to 5 × 10⁻¹) were observed under optimal conditions for two strains, while the third showed 1000-fold lower frequency.
  • Lower temperatures reduced transfer frequencies, but conjugation was still detected across all tested temperatures.
  • Nutrient-poor media (synthetic wastewater, soil extract) significantly decreased transconjugant numbers, with only one strain showing detectable transfer.

Conclusions:

  • Plasmid transfer between microorganisms can occur in natural environments even under suboptimal conditions.
  • Low nutrient availability is the primary factor limiting the environmental dissemination of the IncP-1 plasmid pKJK5.