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Scope  This study aimed to analyse the prevalence, antibiotic resistance and genetic relatedness of  P .  aeruginosa  isolates obtained from potable and recreational water samples (n. 8,351) collected from different settings (swimming pools, n. 207; healthcare facilities, n 1,684; accommodation facilities, n. 1,518; municipal waterworks, n. 4,500; residential buildings, n. 235). Possible mechanisms underlying resistance to imipenem, with particular focus on those involving  opr D-based uptake, were also explored.    Methods and results  Isolation and identification of  Pseudomonas aeruginosa  was performed according to the standardized procedure UNI EN ISO 16266:2008 followed by PCR confirmation. Antibiotic Susceptibility testing was conducted according to EUCAST standardized disk diffusion method. Genetic relatedness of strains was carried out by RAPD. The sequence of the  opr D gene was analyzed by standard method. Fifty-three samples (0.63%) were positive for  P .  aeruginosa , of which 10/207 (4.83%) were from swimming pools. Five isolates (9.43%) were resistant to imipenem, one to Ticarcillin + Clavulanate, one to both Piperacillin and Ticarcillin + Clavulanate. The highest isolation rate of imipenem resistant  P .  aeruginosa  was observed in swimming pool water. Identical RAPD profiles were found in isolates from the same location in the same year or even in different years.    Conclusions  Imipenem resistant strains were identified as carbapenemase-negative and resistance has been associated with inactivating mutations within the  opr D gene, with a concomitant loss of porin. RAPD results proved that a water system can remain colonized by one strain for long periods and the contamination may be difficult to eradicate.  This study has revealed the presence of  P .  aeruginosa  in different water samples, including resistant strains, especially in swimming pools, and confirmed the role of porins as a contributing factor in carbapenem resistance in Gram-negative bacteria.

plos one

Prevalence and antibiotic resistance of Pseudomonas aeruginosa in water samples in central Italy and molecular characterization of oprD in imipenem resistant isolates