Antibiotic Resistance of E. coli Isolated From a Constructed Wetland Dominated by a Crow Roost, With Emphasis on ESBL and AmpC Containing E. coli

Abstract

Information on the dissemination of antibiotic resistance mechanisms in the environment as well as wild life is needed in North America. A constructed wetland (where ∼15,000 American crows roost) was sampled on the University of Washington Bothell Campus for the presence of antibiotic resistant E. coli (ARE). Crow droppings from individual birds and grab samples of water were collected in 2014–2015. E. coli were isolated by selective agar plating. The most frequent antibiotic resistance (AR) of the fecal isolates was to ampicillin (AMP) (53%), followed by amoxicillin-clavulanic acid (AMC) (45%), streptomycin (S) (40%), and nalidixic acid (NA) (33%). Water isolates had similar AR pattern and ∼40% were multidrug resistant. Isolates from water samples collected during storm events showed higher resistance than isolates from no rain days to tetracycline, AMP, AMC, NA, and gentamycin. Extended spectrum beta lactamase (ESBL) containing E. coli with the bla ctx-M was found in three water and nine fecal isolates while bla cmy-2 in 19 water and 16 fecal isolates. Multilocus Sequence Typing analysis (MLST) yielded 13 and 12 different sequence types (STs) amongst fecal and water isolates, many of which could be correlated to livestock, bird, and humans. MLST identified ESBL E. coli belonging to the clinically relevant ST131 clone in six fecal and one water isolate. Three STs found in feces could be found in water on the same dates of collection but not subsequently. Thus, the strains do not appear to survive for long in the wetland. Phylogenetic analysis revealed similar distribution of the water and fecal isolates among the different phylo-groups, with the majority belonging to the commensal B1 phylo-group, followed by the pathogenic B2 phylo-group. This study demonstrates that corvids can be reservoirs and vectors of ARE and pathogenic E. coli , posing a significant environmental threat.

American Crows as Carriers of Extra Intestinal Pathogenic E. coli and Avian Pathogenic-Like E. coli and Their Potential Impact on a Constructed Wetland

“…Crow feces and water samples were collected simultaneously, during 2014–2015, from the crow roost areas within the UW Bothell/Cascadia College (UWB/CC) campus, and details of the sample collection and isolation of E. coli can be found in Sen et al [ 19 ]. Water was collected again from June 2016–April 2017 from the SW8, SW2, NC6, RS1, and RS2 sites before (a day that was preceded by 72 h of no rain) and after rainfall ( Figure 1 ).…”

“…Approximately 100 mg of fecal sample was diluted in 500 μL Phosphate Buffered Saline until homogenous suspension was obtained. An aliquot of 10–20 μL of the suspension was directly plated onto Eosin Methylene Blue (EMB) agar and incubated at 37 °C for 24 h. Colonies with metallic green sheen were isolated as putative E. coli and further verified as described earlier [ 19 ]. Four isolates from each sample were stored at −70 °C in Tryptic Soy Broth (containing 16–20% glycerol until further testing.…”

“…We recently reported 70% of the E. coli isolated from water samples and 60% from the crow fecal samples, obtained from the wetlands of University of Washington Bothell/Cascadia College (UWB/CC) campus (where more than 15,000 crows roost in the autumn and winter months), were resistant to at least one antibiotic [ 19 ]. About 40% of these were multidrug resistant (MDR) as they were resistant to ≥three classes of drugs.…”

“…Extended spectrum beta lactamase (ESBL) containing E. coli were isolated from three water ( n = 184) and nine fecal ( n = 98) isolates. Six of the fecal and one water isolate belonged to the highly virulent, MDR, and internationally distributed sequence type ST131 by Multi Locus Sequence typing analysis [ 19 ]. ST131 is usually a human ExPEC strain.…”

AKTIVITAS EKSTRAK METANOL DAUN ULIN (EUSIDEROXYLON ZWAGERI TEIJSM & BINN) TERHADAP EMPAT JENIS BAKTERI PATOGEN

“…Saat ini penggunaan antibiotik modern menghadapi fenomena resistensi terhadap antibiotik modern yang ditunjukkan oleh beberapa bakteri patogen seperti Escherichia coli (Sen et al 2019). Keberadaan enzim β-lactamase yang dimiliki oleh E. coli menyebabkan bakteri ini resisten terhadap penisilin, cepalosporin dan monobactam aztreonam (Pormohammad et al 2019).…”