Veterinary Hospital Dissemination of CTX-M-15 Extended-Spectrum Beta-Lactamase-Producing Escherichia coli ST410 in the United Kingdom

We characterized extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR) in 32 Escherichia coli extended spectrum cephalosporin (ESC)-resistant clinical isolates from UK companion animals from several clinics. In addition, to investigate the possible dissemination of ESBL clinical isolates within a veterinary hospital, two ESBL-producing E. coli isolates from a dog with septic peritonitis and a cluster of environmental ESC-resistant E. coli isolates obtained from the same clinic and during the same time period, as these two particular ESBL-positive clinical isolates, were also included in the study. Molecular characterization identified bla(CTX-M) to be the most prevalent gene in ESC-resistant isolates, where 66% and 27% of clinical isolates carried bla(CTX-M-15) and bla(CTX-M-14,) respectively. The only PMQR gene detected was aac(6')-Ib-cr, being found in 34% of the ESC E. coli isolates and was associated with the carriage of bla(CTX-M-15). The clinical and environmental isolates investigated for hospital dissemination had a common ESBL/AmpC phenotype, carried bla(CTX-M-15), and co-harbored bla(OXA-1,) bla(TEM-1,) bla(CMY-2,) and aac(6')-Ib-cr. Multilocus sequence typing identified them all as ST410, while pulse-field gel electrophoresis demonstrated 100% homology of clinical and environmental isolates, suggesting hospital environmental dissemination of CTX-M-15–producing E. coli ST410

Biofilm forming in S. Pseudintermedius isolates from atopic dogs

S. pseudintermedius is the main colonizer of skin in dogs, with increased adherence in the ones suffering from atopic dermatitis. It is also the main pathogen involed in pyodermas and has an important zoonotic potential. Biofilm forming was tested using the microtitre plate protocol for 50 S. pseudintermedius sampled from the same number of atopic dogs. Out of these 50 isolates 40 (80%) were methicillin susceptible and 10 (20%) methicillin resistant. S. pseudintermedius isolates sampled from atopic dogs without pyoderma simptoms did not manifest an important biofilm production. Our results show that 60% of all isolates were not biofilm producers, 38% were weak biofilm producers and only one isolate was a moderate biofilm producer. Adherence capability did not show significand differences between MSSPs (methicillin susceptible S. pseudintermedius) and MRSPs (methicillin resistant S. pseudintermedius)

Clonal Dissemination of Extended-Spectrum Cephalosporin-Resistant Enterobacterales between Dogs and Humans in Households and Animal Shelters of Romania

Faecal carriage of extended-spectrum cephalosporin-resistant (ESC-R) Enterobacterales in healthy pets is a concerning issue. This study aimed to determine the prevalence, genetic background, and potential for interspecies transmission of these bacteria between dogs and humans within the same household (HH) or shelter environment in Romania. Faecal samples (n = 263) collected from healthy dogs (n = 102), their owners (n = 32), as well as dogs (n = 110) and staff (n = 19) from dog shelters, were screened for ESC-R carriage. Clonal relatedness of canine and human Escherichia coli isolates was established using Fourier Transform Infrared Spectroscopy (FTIR), followed by Illumina WGS of selected isolates. The highest prevalence of ESC-R Enterobacterales faecal carriage was identified in staff working at dog shelters (78.9%), followed by dogs from households (44.11%), dog owners (43.7%), and dogs from shelters (27%). FTIR identified 15 clusters of closely related E. coli isolates, including dog and human isolates from the same environment. Co-carriage of ESC-R isolates in both the dog and owner was identified in 12 HHs (37.5%), with two HHs (6%) having both the owner and dog carrying isolates with identical FTIR spectra, phylogroup, resistance genes, and Inc plasmids. Major ExPEC lineages such as ST127, ST10, ST155, and ST88 were detected in human and dog isolates. Our study revealed a high prevalence of faecal ESC-R E. coli carriage in both dogs and humans from Romanian households and shelters, where bidirectional clonal transmission between humans and dogs is likely. Furthermore, we identified ESC-R Enterobacterales co-carriage in people and dogs sharing the same environment using FTIR, demonstrating its value in AMR surveillance for humans and animals

Investigation of In Vitro Susceptibility and Resistance Mechanisms in Skin Pathogens: Perspectives for Fluoroquinolone Therapy in Canine Pyoderma

Fluoroquinolones (FQ) are commonly used in dogs with bacterial skin infections. Their use as first choice, along with the increased incidence of FQ-resistance, represents a risk to animal and public health. Our study determined minimum inhibitory (MIC) and bactericidal (MBC) concentrations of five FQs in Staphylococcus aureus, Staphylococcus pseudintermedius, and Escherichia coli, together with FQ-resistance mechanisms. MICs, efflux pump (EP) overexpression and MBCs were measured in 249 skin infection isolates following CLSI guidelines (CLSI VET01-A4, CLSI M26-A). Chromosomal and plasmid-mediated resistance genes were investigated after DNA extraction and sequencing. FQ-resistance was detected in 10% of methicillin-susceptible (MS), 90% of methicillin-resistant (MR) staphylococci and in 36% of E. coli. Bactericidal effect was observed except in 50% of MRSA/P for ciprofloxacin and in 20% of MRSPs for enrofloxacin. Highest MICs were associated with double mutation in gyrA (Ser83Leu + Asp87Asn), efflux pumps and three PMQR genes in E. coli, and grlA (Ser80Phe + Glu84Lys) in S. aureus. EP overexpression was high among E. coli (96%), low in S. aureus (1%) and absent in S. pseudintermedius. Pradofloxacin and moxifloxacin showed low MICs with bactericidal effect. Since in vitro FQ resistance was associated with MR, FQ use should be prudently guided by susceptibility testing

Are the Hands of Veterinary Staff a Reservoir for Antimicrobial-Resistant Bacteria? A Randomized Study to Evaluate Two Hand Hygiene Rubs in a Veterinary Hospital

Hand hygiene (HH) is the most successful intervention for hospital infection control. HH rubs with residual action are desired. This study aimed to compare the efficacy of alcohol (A-HH) and lactic acid (LA-HH) rubs, with the latter being marketed as having residual activity. We investigated reductions in bacterial colony-forming units (CFUs), prevalence of antimicrobial-resistant (AMR) organisms, and risk factors for increased counts on the hands of veterinary staff. A randomized, crossover study (53 individuals) was performed in a referral veterinary teaching hospital. Hand plates were taken before, immediately after, and 6 hours after HH. A blinded investigator counted CFUs per plate. Methicillin-resistant Staphylococcus aureus/pseudintermedius (MRSA/MRSP), Enterobacteriaceae, and Pseudomonas species (spp.) were characterized. Gender, profession, time point, and HH product were included as variables within multivariable analyses. A significant reduction in bacterial CFU was seen immediately after A-HH rub application (p < 0.001); however, neither product showed any significant residual action. Veterinarians had higher bacterial CFUs than nurses (p = 0.005); contact with patients, rather than the environment, was also associated with higher counts (p < 0.001). MRSA, MRSP, Enterobacteriaceae spp., and Pseudomonas spp. were detected on 7%, 2%, 14%, and 2% of study participant's hands (n = 208 samples), respectively. Frequent HH administration using an A-HH rub was effective at reducing bacterial CFU on hands in vivo in this veterinary hospital setting, but its use needs further encouragement in veterinary staff. The high prevalence of antimicrobial bacteria on hands is of concern; they might act as reservoirs for patients, the environment, and in-contact people

Rapid typing of Klebsiella pneumoniae and Pseudomonas aeruginosa by Fourier-transform Infrared spectroscopy informs infection control in veterinary settings

IntroductionThe emergence of multi-drug resistant (MDR) pathogens linked to healthcare-associated infections (HCAIs) is an increasing concern in modern veterinary practice. Thus, rapid bacterial typing for real-time tracking of MDR hospital dissemination is still much needed to inform best infection control practices in a clinically relevant timeframe. To this end, the IR Biotyper using Fourier-Transform InfraRed (FTIR) spectroscopy has the potential to provide fast cluster analysis of potentially related organisms with substantial cost and turnaround time benefits.Materials and methodsA collection of MDR bacterial isolates (n = 199, comprising 92 Klebsiella pneumoniae and 107 Pseudomonas aeruginosa) obtained from companion animal (i.e., dogs, cats and horses) clinical investigations, faecal and environmental screening from four veterinary facilities between 2012 and 2019 was analysed retrospectively by FTIR spectroscopy. Its performance was compared against MLST extracted from whole genomes of a subset of clustering isolates (proportionally to cluster size) for investigation of potential nosocomial transmission between patients and the surrounding hospital environments.ResultsConcordance between the FTIR and MLST types was overall high for K. pneumoniae (Adjusted Rand Index [ARI] of 0.958) and poor for P. aeruginosa (ARI of 0.313). FTIR K. pneumoniae clusters (n = 7) accurately segregated into their respective veterinary facility with evidence of intra-hospital spread of K. pneumoniae between patients and environmental surfaces. Notably, K. pneumoniae ST147 intensely circulated at one Small Animal Hospital ICU. Conversely, Pseudomonas aeruginosa FTIR clusters (n = 18) commonly contained isolates of diversified hospital source and heterogeneous genetic background (as also genetically related isolates spread across different clusters); nonetheless, dissemination of some clones, such as P. aeruginosa ST2644 in the equine hospital, was apparent. Importantly, FTIR clustering of clinical, colonisation and/or environmental isolates sharing genomically similar backgrounds was seen for both MDR organisms, highlighting likely cross-contamination events that led to clonal dissemination within settings.ConclusionFTIR spectroscopy has high discriminatory power for hospital epidemiological surveillance of veterinary K. pneumoniae and could provide sufficient information to support early detection of clonal dissemination, facilitating implementation of appropriate infection control measures. Further work and careful optimisation need to be carried out to improve its performance for typing of P. aeruginosa veterinary isolates.</jats:sec