Characterization of cefotaxime-resistant urinary Escherichia coli from primary care in South-West England 2017–18

Abstract

Objectives Third-generation cephalosporin-resistant Escherichia coli from community-acquired urinary tract infections are increasingly reported worldwide. We sought to determine and characterize the mechanisms of cefotaxime resistance employed by urinary E. coli obtained from primary care, over 12 months, in Bristol and surrounding counties in South-West England. Methods Cefalexin-resistant E. coli isolates were identified from GP-referred urine samples using disc susceptibility testing. Cefotaxime resistance was determined by subsequent plating onto MIC breakpoint plates. β-Lactamase genes were detected by PCR. WGS was performed on 225 isolates and analyses were performed using the Center for Genomic Epidemiology platform. Patient information provided by the referring general practices was reviewed. Results Cefalexin-resistant E. coli (n=900) isolates were obtained from urines from 146 general practices. Following deduplication by patient approximately 69% (576/836) of isolates were cefotaxime resistant. WGS of 225 isolates identified that the most common cefotaxime-resistance mechanism was blaCTX-M carriage (185/225), followed by plasmid-mediated AmpCs (pAmpCs) (17/225), AmpC hyperproduction (13/225), ESBL blaSHV variants (6/225) or a combination of both blaCTX-M and pAmpC (4/225). Forty-four STs were identified, with ST131 representing 101/225 isolates, within which clade C2 was dominant (54/101). Ciprofloxacin resistance was observed in 128/225 (56.9%) of sequenced isolates, predominantly associated with fluoroquinolone-resistant clones ST131 and ST1193. Conclusions Most cefalexin-resistant E. coli isolates were cefotaxime resistant, predominantly caused by blaCTX-M carriage. The correlation between cefotaxime resistance and ciprofloxacin resistance was largely attributable to the high-risk pandemic clones ST131 and ST1193. Localized epidemiological data provide greater resolution than regional data and can be valuable for informing treatment choices in the primary care setting.

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

“…The fourth plasmid (pIV_IncFII_DHA) was 83,429 bp in size and belonged to the IncFII plasmid family. pIV_IncFII_DHA showed high score with both the p133355_SW_C4_Cam-1 plasmid (81,724 bp) reported in 2019 in a Citrobacter amalonaticus collected from a human stool specimen in Switzerland (CP041363.1; 99.64% sequence identity, 97% sequence coverage), and with the pUB_DHA-1 plasmid (81,754 bp), present in an E. coli strain from a human urine sample in United Kingdom (MK048477; 99.55% sequence identity, 97% query coverage) (Findlay et al, 2019). The plasmid backbone carried regions responsible for replication (repA), conjugative transfer system (tra, yhfA, finO genes), maintenance (parM) and toxin-antitoxin (TA) systems (mazEF/mazEF).…”

Expansion of a subset within C2 clade of Escherichia coli sequence type 131 (ST131) is driving the increasing rates of Aminoglycoside resistance: a molecular epidemiology report from Iran

“…Virotype C is considered to be the most widely distributed virotype among ST131, occurring in all ST131 clades ( 17 ). Here, virotype C was predominant among clade C1, specifically C1-M27 subclade, as also reported recently from South-West England and Europe ( 17, 18 ).…”

“…Virotype C is considered to be the most widely distributed virotype among ST131, occurring in all ST131 clades ( 17 ). Here, virotype C was predominant among clade C1, specifically C1-M27 subclade, as also reported recently from South-West England and Europe ( 17, 18 ). In contrast to several European studies which reported the virotype A as the dominated virotype among C2 clade ( 12 ), in our study virotypes E and F were found as the most common virotypes, representative of > 60% of the C2 clade strains (22 out of 34 strains).…”

Resistance to Ceftazidime/Avibactam Plus Meropenem/Vaborbactam When Both are Used Together Achieved in Four Steps From Metallo-β-Lactamase NegativeKlebsiella pneumoniae

“…bla CTX-M-14 was amplified from a human urinary E. coli isolated from primary care (38) by PCR with Phusion High-Fidelity DNA Polymerase (NEB, UK) using primers CTX-M-14 FW (5′-CCGGAATTCAATACTACCTTGCTTTCTGA-3′) and CTX-M-14 RV (5′-CCGGAATTCCGTAGCGGAACGTTCATCAG-3′) and ligated into pUBYT (39) at the EcoRI site. CTX-M-14 site-directed mutagenesis ( bla CTX-M-14-P170S ) was performed using the QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, USA) with the primer CTX-M-14-P170S-FW (5′-TCTGGATCGCACTGAATCTACGCTGAATACCGC-3′).…”