The Influence of Aquaculture Effluents on the Prevalence and Biocides Resistance of Opportunistic Pseudomonas fluorescens Bacteria in the Drwęca River Protected under the Natura 2000 Network

Abstract

The aim of this study was to determine the effect of trout aquaculture effluents on the Drwęca River. The count of opportunistic pathogen Pseudomonas fluorescens (OPPF) in the total Pseudomonas fluorescens population (TPFP) were determined by plating on King B medium and fluorescence in situ hybridization. The resistance of OPPF strains to 12 antibiotics and two disinfectants was evaluated. Significant differences (p ≤ 0.05) in OPPF counts were found between seven sampling sites. OPPF counts were highest in samples collected directly downstream from three fish farms. More than 50% of these isolates demonstrated multiple-drug resistance to ampicillin, mezlocillin, cefotaxime, norfloxacin, tetracycline and two disinfectants (Steridial and chloramine T). Of these, 52% were resistant to high doses of cefotaxime and norfloxacin (MIC ≥ 256 µg·mL−1), and 65% were resistant to the maximum doses of Steridial (MIC Ste ≥ 25 mL·m−3) and chloramine T (MIC Chlor ≥ 20 mg·L−1). All OPPF sampled upstream from the farms were sensitive to low concentrations of CTX (cefotaxime) and NOR (norfloxacin) (MIC ≤ 2 µg·mL−1), Steridial (MIC Ste ≤ 5 mL·m−3) and chloramine T (MIC Chlor ≤ 2.5 mg·L−1). Agglomerative clustering revealed two clusters: strains from samples collected upstream and downstream from trout farms. The results indicate that aquaculture effluents significantly affect the prevalence of biocides resistant OPPF along the river continuum.