Deadly Puppy Infection Caused by an MDR Escherichia coli O39 blaCTX–M–15, blaCMY–2, blaDHA–1, and aac(6)-Ib-cr – Positive in a Breeding Kennel in Central Italy

Abstract

Antimicrobial consumption in veterinary medicine has led to the spread of multi drug-resistance in clinically important bacteria, with the companion animals and their environment involved as emerging reservoirs. While CTX-M-15 and CMY-2 acquired β-lactamases have been widely detected in the bacterial population of companion and breeding animals in European area, DHA-1 enzymes have been rarely reported in veterinary medicine. The aim of the study was to characterize the Escherichia coli associated with mortality of a litter of Bulldog puppies in a breeding kennel located in Pesaro area, Central Italy. The E. coli strains O39 serotype were resistant to 3rd/4th generation cephalosporins, chloramphenicol, aminoglycosides, trimethoprimsulfamethoxazole, and ciprofloxacin, retaining susceptibility to carbapenems, colistin, fosfomycin, and levofloxacin (by Microscan Autoscan4, EUCAST clinical breakpoints). Pulse field gel electrophoreses (PFGE-XbaI) on five E. coli strains revealed the presence of a single profile. Whole genome sequencing (WGS) analysis revealed a complex resistome, harboring bla TEM−1b , bla CTX−M−15 , bla OXA−1 , aph(6)-Ib, aac(6)Ib-cr, aac(3)-Ila, aph(6)-Id, aadA1, qnrB1, sul2, catA1, catB3, tetA, and dfrA14 genes located on a 302597 bp IncHI2/HI2A plasmid. Moreover, bla DHA−1 , qnrB4, mph(A), sul1, and dfrA17 determinants were carried on an 83,429 bp IncFII plasmid. A bla CMY−2 determinant was carried on a 90,249 bp IncI1 plasmid. Two IncX1 and IncX4 plasmids without antimicrobial resistance genes were also detected. The presence of lpfA, iss, astA, and gad virulence factors was highlighted. This is the first report in Italy on an invasive infection in eight 2-weeks old dogs caused by the same MDR E. coli O39 bla CTX−M−15 , bla CMY−2 , bla DHA−1 , and aac(6)-Ib-cr positive strain. The above MDR E. coli clone caused the death of the entire litter, despite amoxicillin-clavulanate and enrofloxacin administration. The tank for storage of the water used to prepare the milk-based meal for the litter was the suspected reservoir.

Circulation of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli of Pandemic Sequence Types 131, 648, and 410 Among Hospitalized Patients, Caregivers, and the Community in Rwanda

“…Finally, we demonstrate that similar plasmid sequences were present in strains from different sample groups, thus likely indicating mobile genetic element transmission, and underlining the importance of plasmid-driven spread of antimicrobial resistance independent of the host's phylogenetic background (Schaufler et al, 2016a;Ranjan et al, 2018). Interesting in addition to similarities among strains from the Rwandan setting is in particular the close relationship to an external plasmid, which has been obtained only recently (Marchetti et al, 2020). This highlights the sometimes global spread of such mobile genetic elements and their bacterial hosts.…”

“…We also compared some of our plasmid sequences of representative strains (criteria: top BLAST-hits) against a publicly available reference plasmid pIV_IncHI2_CTX_M_15 (Marchetti et al, 2020; Supplementary Figure 3). This bla CTX−M−15containing plasmid has been originally obtained from an E. coli ST58 strain causing a deadly puppy infection in Italy in 2019.…”

Whole genome sequence analysis of ESBL-producing Escherichia coli recovered from New Zealand freshwater sites

“…This IncHI2 plasmid has been isolated from a variety of Enterobacteriaceae species including the Enterobacter cloacae complex, Salmonella enterica , Klebsiella pneumoniae , Citrobacter freundii and E. coli [78–81]. It has been suggested that the reason for the successful spread of this IncHI2 plasmid is because it harbours numerous metal resistance genes [80].…”

Enrofloxacin—The Ruthless Killer of Eukaryotic Cells or the Last Hope in the Fight against Bacterial Infections?

“…AAC(6′)-Ib-cr has two amino acid changes, Trp102Arg and Asp179Tyr, which together are necessary for the enzyme’s ability to acetylate the antibiotic [ 103 ]. The presence of a gene encoding AAC(6′)-Ib-cr was confirmed in clinical enrofloxacin-resistant strains of E. coli [ 104 ] and S. Indiana [ 105 ].…”