This study aimed to define the frequency of resistance to critically important antimicrobials (CIAs) [i.e. extended-spectrum cephalosporins (ESCs), fluoroquinolones (FQs) and carbapenems] among Escherichia coli isolates causing clinical disease in Australian food-producing animals. Clinical E. coli isolates (n=324) from Australian food-producing animals [cattle (n=169), porcine (n=114), poultry (n=32) and sheep (n=9)] were compiled from all veterinary diagnostic laboratories across Australia over a 1-year period. Isolates underwent antimicrobial susceptibility testing to 18 antimicrobials using the Clinical and Laboratory Standards Institute disc diffusion method. Isolates resistant to CIAs underwent minimum inhibitory concentration determination, multilocus sequence typing (MLST), phylogenetic analysis, plasmid replicon typing, plasmid identification, and virulence and antimicrobial resistance gene typing. The 324 E. coli isolates from different sources exhibited a variable frequency of resistance to tetracycline (29.0-88.6%), ampicillin (9.4-71.1%), trimethoprim/sulfamethoxazole (11.1-67.5%) and streptomycin (21.9-69.3%), whereas none were resistant to imipenem or amikacin. Resistance was detected, albeit at low frequency, to ESCs (bovine isolates, 1%; porcine isolates, 3%) and FQs (porcine isolates, 1%). Most ESC- and FQ-resistant isolates represented globally disseminated E. coli lineages (ST117, ST744, ST10 and ST1). Only a single porcine E. coli isolate (ST100) was identified as a classic porcine enterotoxigenic E. coli strain (non-zoonotic animal pathogen) that exhibited ESC resistance via acquisition of blaCMY-2. This study uniquely establishes the presence of resistance to CIAs among clinical E. coli isolates from Australian food-producing animals, largely attributed to globally disseminated FQ- and ESC-resistant E. coli lineages.

Unlike Escherichia coli strains belonging to phylogroup B2, the clinical significance of strains belonging to phylogroup F is not well understood. Here we report on a collection of phylogroup F strains recovered in Australia from faeces and extra-intestinal sites from humans, companion animals and native animals, as well as from poultry meat and water samples. The distribution of sequence types was clearly non-random with respect to isolate source. The antimicrobial resistance and virulence trait profiles also varied with the sequence type of the isolate. Phylogroup F strains tended to lack the virulence traits typically associated with phylogroup B2 strains responsible for extra-intestinal infection in humans. Resistance to fluoroquinolones and/or expanded-spectrum cephalosporins was common within ST648, ST354 and ST3711. Although ST354 and ST3711 are part of the same clonal complex, the ST3711 isolates were only recovered from native birds being cared for in a single wildlife rehabilitation centre, whereas the ST354 isolates were from faeces and extra-intestinal sites of dogs and humans, as well as from poultry meat. Although ST354 isolates from chicken meat in Western Australia were distinct from all other ST354 isolates, those from poultry meat samples collected in eastern Australia shared many similarities with other ST354 isolates from humans and companion animals.

Background: Extraintestinal infections caused by multidrug-resistant (MDR) Escherichia coli and Enterobacter are becoming more common in veterinary medicine. Objective: To generate hypotheses for risk factors for dogs acquiring extraintestinal infection caused by MDR E. coli and Enterobacter, describe antimicrobial resistance profiles and analyze treatment and clinical outcomes. Animals: Thirty-seven dogs diagnosed with extraintestinal infection caused by MDR E. coli and Enterobacter spp. between October 1999 and June 2006. Methods: Retrospective case series assembled from hospital records data, including clinical history before 1st MDR isolation and treatment outcome. Identity and antimicrobial susceptibility profiles were confirmed by standard microbiological techniques for 57 isolates. Results: Most dogs had an underlying disease condition (97%), received prior antimicrobial treatment (87%), were hospitalized for ≥3 days (82%), and had a surgical intervention (57%). The urinary tract was the most common infection site (62%), and urinary catheterization, bladder stasis, or both were common among dogs (24%). Some dogs were treated with high doses of co-amoxyclavulanate (n = 14) and subsequently recovered even though the isolates showed in vitro resistance to this antimicrobial. Other dogs were successfully treated with chloramphenicol (n = 11) and imipenem (n = 2). Conclusion and Clinical Importance: Predisposing disease condition, any prior antimicrobial use rather than a specific class of antimicrobial, duration of hospitalization, and type of surgical procedure might be risk factors for acquiring MDR extraintestinal infections. Whereas culture and sensitivity results can indicate use of last-resort antimicrobials such as imipenem for MDR infections, some affected dogs can recover after administration of high doses of co-amoxyclavulanate.

The genetic determinants involved in reduced susceptibility to third-generation cephalosporins and aztreonam were identified in ten canine Enterobacter isolates associated with opportunistic infections in three veterinary hospitals in Brisbane, Australia. All isolates were evaluated by a combination of phenotypic (broth microdilution and disc susceptibility, modified disc diffusion and IEF) and genotypic (PFGE, plasmid analysis, Southern blot hybridization, bacterial conjugation, PCR and sequencing) methods to investigate genetic relatedness and to identify plasmid-mediated resistance genes, in particular β-lactamase genes responsible for extended-spectrum cephalosporin resistance. The ten canine isolates were genotypically diverse based on PFGE and belonged to either Enterobacter cloacae or Enterobacter hormaechei on the basis of 16S rRNA gene sequence analysis. Plasmid profiles were also diverse. Nine isolates contained a transmissible bla_SHV-12-carrying plasmid (∼140 kb) that also conferred resistance to chloramphenicol, gentamicin, spectinomycin, tetracycline, trimethoprim and sulfonamides. In all plasmid-mediated extended-spectrum β-lactamase (ESBL)-producing isolates including transconjugants, bla_SHV-12 was shown to reside in a ∼6.5 kb plasmid fragment. The remaining isolate that was not an ESBL producer possessed an AmpC β-lactamase gene (bla_CMY-2) on a ∼93 kb transmissible plasmid. This plasmid did not contain any other antimicrobial resistance genes. Additional plasmid-mediated β-lactamases identified in some isolates included bla_TEM and bla_OXA-10. This is the first report of canine Enterobacter isolates containing transmissible plasmid-mediated bla_SHV-12 and bla_CMY-2 resistance genes. Therefore, Enterobacter isolated from opportunistic infections in dogs may be an important reservoir of plasmid-mediated resistance genes, which could potentially be spread to other members of the Enterobacteriaceae.