Abstract
Asymptomatic colonization with extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae has been described for humans, various mammal species, and birds. Here, antimicrobial resistant bacteria were recovered from dog feces originating in Germany, Kosovo, Afghanistan, Croatia, and Ukraine, with a subset of mostly E. coli isolates obtained from a longitudinal collection over twelve months. In vitro antimicrobial resistance testing revealed various patterns of resistance against single or all investigated beta-lactam antibiotics, with none of the 101 isolates resistant against two tested carbapenem antibiotics. Whole genome sequence analysis revealed bacteria species-specific patterns for 23 antimicrobial resistance coding DNA sequences (CDS) that were unapparent from the in vitro analysis alone. Phylogenetic analysis of single nucleotide polymorphisms (SNP) revealed clonal bacterial isolates originating from different dogs, suggesting transmission between dogs in the same community. However, individual resistant E. coli clones were not detected over a period longer than seven days. Multi locus sequence typing (MLST) of 85 E. coli isolates revealed 31 different sequence types (ST) with an accumulation of ST744 (n = 9), ST10 (n = 8), and ST648 (n = 6), although the world-wide hospital-associated CTX-M beta-lactamase producing ST131 was not detected. Neither the antimicrobial resistance CDSs patterns nor the phylogenetic analysis revealed an epidemiological correlation among the longitudinal isolates collected from a period longer than seven days. No genetic linkage could be associated with the geographic origin of isolates. In conclusion, healthy dogs frequently carry ESBL-producing bacteria, independent to prior treatment, which may be transmitted between individual dogs of the same community. Otherwise, these antimicrobial resistant bacteria share few commonalities, making their presence eerily unpredictable.
Highlights
Beta-lactams are among the most popular antibiotics, worldwide, for the treatment of bacterial infections [1]
A high prevalence of extended-spectrum betalactamases (ESBL)-producing bacteria is suspected in livestock, our findings were surprising considering that the investigated animals were clinically healthy and untreated [42]
Concerning was the result that 9% of the characterized E. coli isolates from clinically healthy German dogs were completely resistant against all tested cephalosporins (COX, CTX, CAZ, and CEP) (S2 Table)
Summary
Beta-lactams are among the most popular antibiotics, worldwide, for the treatment of bacterial infections [1]. ESBL-producing Enterobacteriaceae have been identified from a plethora of sources, including humans, animals, food, feed, and other environmental sources [3,4,5,6,7]. ESBL-producing Escherichia coli isolated from dogs were first described in 1988, following treatment of the dogs with beta-lactam antibiotics [8]. The presence of ESBL-producing bacteria has been described repeatedly for sick, and completely healthy companion animals, including dogs [4,9,10,11,12,13]. One longitudinal study occurring over six months identified a variety of ESBL-producing Enterobacteriaceae in healthy dogs with highly dynamic fecal shedding patterns, occurring either continuously or periodically [14]
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