BACKGROUND:Gram-negative ESKAPE organisms are an emerging threat in veterinary medicine that also have zoonotic potential. However, little is known about the prevalence and risks of these organisms in pet food. We present the incidental finding of gram-negative ESKAPE organisms within samples of raw meat diets for dogs in the United Kingdom. METHODS:Samples of commercially available frozen raw-meat diets for dogs (n = 110) underwent bacterial culture and antimicrobial susceptibility testing. Bacterial species were identified via matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. Sequence types and resistance genes were determined by whole-genome sequencing. RESULTS:Antimicrobial-resistant Acinetobacter baumannii complex, Pseudomonas aeruginosa and Enterobacter spp. were isolated from eight, 10 and four samples, respectively. Isolates demonstrated phenotypic resistance to highest-priority critically important antibiotics, including meropenem (P. aeruginosa). Key acquired resistance genes were identified. LIMITATIONS:The microbiological methods used may not have been optimal for isolation of Gram-negative ESKAPE organisms. CONCLUSION:The findings of this study further highlight the potential One Health risks associated with feeding dogs raw-meat diets.

Introduction Phaeohyphomycosis is an opportunistic infection caused by melanized fungi, primarily affecting immunocompromised hosts. Accurate identification is challenging due to morphological similarities among species, often leading to misidentification. Case presentation A 7.6-year-old immunocompromised Shetland Sheepdog, receiving prednisolone and ciclosporin for sterile nodular panniculitis, presented with a solitary purulent subcutaneous nodule on the right flank. Histopathology of an excisional biopsy revealed pyogranulomatous inflammation with pigmented septate fungal hyphae, consistent with phaeohyphomycosis. Fungal culture produced black, powdery colonies within four days incubation. Initial morphological identification suggested Exophiala spp., later revised to Verruconis gallopava by a reference laboratory. Definitive identification was achieved by sequencing of the internal transcribed spacer (ITS) and β-tubulin gene regions, confirming Scolecobasidium dracaenae. Antifungal susceptibility testing, performed according to EUCAST guidelines, demonstrated high minimum inhibitory concentrations (MICs >8 mg/L) to all agents tested. Conclusion This case highlights the limitations of conventional identification methods and the importance of molecular diagnostics for rare fungal pathogens. Despite in vitro antifungal resistance, successful resolution was achieved with surgical management and topical therapy. To the authors’ knowledge, this represents the first reported canine infection caused by S. dracaenae.

Introduction: Antimicrobial resistance (AMR) is a globally relevant public health priority requiring coordinated One Health responses. In recent years, the emergence of novel AMR phenotypes and increasing AMR trends in bacteria from companion animal and horses have become an apparent concern worldwide; nevertheless, limited national resistance monitoring programmes in Europe systematically collect and analyse data from companion animal and equine infections. In the UK, private veterinary microbiology diagnostic laboratories hold valuable AMR data, however, there is a lack of alignment in the methodologies used by diagnostic laboratories to generate these data. Gap Statement: Substantial knowledge gaps exist in the current landscape of veterinary microbiological diagnostic testing approaches for AMR in companion animals and horses. The VetCLIN AMR initiative aims to engage with UK veterinary diagnostic laboratories to capture methods used for generating routine antimicrobial susceptibility data which impacts on national AMR surveillance. Aim: To perform a UK-wide survey of veterinary laboratories processing companion animal and equine specimens evaluating the standard practices used for microbiology testing with a focus on AMR screening. Methodology: An online survey targeting veterinary diagnostic laboratories in the UK processing clinical specimens from companion animals and horses. Results: Twenty-one (n=21) UK veterinary microbiology laboratories responded, comprising commercial (10/21), university (7/21), and in-house veterinary practice/hospital laboratories (4/21). Methodological heterogeneity in bacterial identification, antimicrobial susceptibility testing (AST), AMR detection and reporting was identified across UK veterinary laboratories. Overall, the laboratories’ preferred AST method was minimum inhibitory concentration (43%), with most of these using the VITEK-1/VITEK-2 system (52%). A combination of CLSI and EUCAST standards was most used by these laboratories (62%) for AST interpretation, with limited adoption of the lower clinical breakpoints for Enterobacterales (amoxicillin–clavulanic acid and fluoroquinolones). Detection of AMR mechanisms was focused on clinical guidance and phenotypes commonly encountered among companion and equine isolates, such as screening for methicillin resistance in staphylococci (95%) and extended-spectrum beta-lactamase (ESBL)-production in Enterobacterales (48%). Up to 30% of UK veterinary laboratories screened for carbapenem resistance in Gram-negative bacteria. Detected AMR phenotypes are infrequently characterised or referred to reference laboratories and, in the case of carbapenem resistance, these are often not reported to clinicians. Conclusion: This study identified a variety of workflows used by veterinary diagnostic laboratories for bacterial identification, methodologies and interpretative criteria used for performing and reporting AST, as well as AMR screening approaches. Several gaps hindering national AMR surveillance in dogs, cats and horses in the UK have become evident. Strengthening surveillance capacity should prioritise consistent screening and characterisation of AMR mechanisms such as ESBL and carbapenemase production, to enable effective responses to AMR emergence and spread in companion animals and equine clinical infections.

Escherichia coli is a commensal intestinal bacterium of humans and animals and is frequently associated with antimicrobial resistance (AMR). This study investigated the phenotypic and genotypic AMR and virulence factors in E. coli isolated from 120 young healthy chickens never treated with antibiotics and coccidiostats. The disk diffusion method employed on all 120 isolates revealed the highest resistance (11.67%) to chloramphenicol and trimethoprim-sulfamethoxazole. All isolates were susceptible to aztreonam and gentamicin, whereas over 95% showed susceptibility to amikacin, cefoxitin, ciprofloxacin, and ertapenem. Minimum inhibition concentration test found 7 (5.83%) isolates resistant to colistin. Eight strains (6.67%) were found to be multidrug-resistant (MDR). Molecular analyses conducted on resistant and intermediate isolates showed that 11/13 penicillin-resistant strains carried the blaTEM gene, 12/12 tetracycline-resistant strains had tetA, 4/14 chloramphenicol-resistant and 1/2 chloramphenicol-intermediate strains had cmlA. The astA gene, potentially indicative of enteroaggregative E. coli (EAEC), was found in 15% of isolates. Virulence genes characterizing other pathotypes investigated were not detected. Chickens reared in optimal conditions and never treated with antibiotics may harbor antimicrobial-resistant and pathogenic bacterial strains that could become a public health hazard. Good farm management and hygiene are fundamental to reducing the circulation of pathogenic strains and the need for antimicrobial treatments.

Raw meat diets for pets are frequently contaminated with potentially pathogenic and zoonotic bacteria. Here we present the first report, to our knowledge, of Bordetella hinzii, a respiratory pathogen of poultry and rare but emerging opportunistic respiratory pathogen in humans, detected incidentally in commercially available frozen pre-prepared raw meat diets for cats. B. hinzii was isolated from five samples of raw meat diets, from two separate brands, purchased from a retailer in the United Kingdom. Samples were a single-protein source and comprised chicken, beef, and rabbit. Whole genome sequencing indicated the presence of antimicrobial resistance genes with potential to confer resistance to multiple drug classes, including classes of highest priority critically important antibiotics. Whilst this study was limited by a small dataset and additional research surrounding the risks of transmission of B. hinzii is required, this novel finding indicates a further potential risk associated with the provision of raw meat diets for pets, particularly for the immunocompromised, that warrants further investigation.

Introduction:Intensive medical care provided in companion animal practices carries the potential risk of selecting and disseminating multidrug-resistant organisms (MDROs). However, data on infection, prevention and control standards specific to small animal veterinary practices (SAVPs) remains limited. The goal of our work was to evaluate the environmental contamination and staff carriage by MDROs in veterinary practices across Portugal. Methods:Fourteen SAVPs were enrolled. Environmental samples were collected from critical areas such as operating room, wards and pre-operative area. Veterinary team members voluntarily gave nasal, hand and rectal swabs. All samples were screened for the presence of, including extended-spectrum β-lactamases (ESBL)- and carbapenemase-producing Gram-negative bacteria and methicillin-resistant Staphylococcus spp. (MRS). Whole-genome sequencing was performed for carbapenem resistant strains. Results:Environmental evaluation by surface swabs revealed that 6.5% (n=32/490) were contaminated with multidrug-resistant Gram-negative bacteria. OXA-23-producing Acinetobacter spp. (n=5) and IMP-8-producing Pseudomonas juntendi (n=2) strains were described on different locations of different SAVPs. Moreover, Stenotrophomonas maltophilia (n=12) and Pseudomonas aeruginosa (n=3) strains were also found on multiple surfaces of different SAVPs. Three human samples (two rectal, one hand) had carbapenem-resistant P. aeruginosa strains by OprD mutations, while S. maltophilia strains were recovered from four samples (two rectal, two hands). One nasal swab was positive for carbapenem-resistant Klebsiella pneumoniae ST11. Only one SAVP surface was positive for the newly typed for methicillin-resistant Staphylococcus aureus (MRSA) ST9220-II. MRSA nasal carriage was found in 14% of samples (n=9/64), with an equal prevalence of ST22-IV and ST8-VI. As for hand samples, MRSA was present in 10.7% (n=4/38), with a predominance of ST8-VI. Discussion:These emerging data indicate that SAVPs may significantly contribute to the dissemination of MDROs. To address this, rigorous infection, prevention and control (IPC) measures should be implemented, alongside educational workshops directed to all veterinary staff as well as to veterinary and nursing students.

Method jBile colonisation occurs with bacteria originating from the gut.Unfortunately, there is a lack of standard procedures to guide processing specimens form companion animals which may lead to variable culture results including missing relevant pathogens.The objective of this study was to compare bacterial culture protocols and results for bile samples from different laboratories aiming to generate a consensus protocol for these specimens. Objectives Results

Antimicrobial stewardship initiatives are widely regarded as a cornerstone for ameliorating the global health impact of antimicrobial resistance. Within companion animal health, such efforts have largely focused on development and dissemination of antimicrobial stewardship guidelines (ASGs). However, there have been few attempts to understand veterinarian attitudes towards and knowledge of ASGs or to determine how awareness regarding ASGs might best be increased. An online survey regarding ASGs was formulated for veterinarians who treat companion animals. The survey was distributed across 46 European and associated countries between 12 January and 30 June, 2022. In total, 2271 surveys were completed, with 64.9% of respondents (n = 1474) reporting awareness and usage of at least one ASG. Respondents from countries with greater awareness of ASGs tended to report more appropriate use of antimicrobials (Spearman's rank coefficient = 0.6084, P ≤ 0.001), with respondents from countries with country-specific ASGs tending to score highest across both awareness and appropriate use domains. Respondents prioritised guidance around antimicrobial choice (82.0%, n = 1863), duration of treatment (66.0%, n = 1499), and dosage (51.9%, n = 1179) for inclusion in future ASGs, with 78.0% (n = 1776) of respondents preferring ASGs to be integrated into their patient management system. Awareness of ASGs and their use in companion animal veterinary practice appears to be greater than previously reported, with respondents tending to report antimicrobial prescription decision making broadly in line with current clinical recommendations. However, further initiatives aimed at maximising accessibility to ASGs both within countries and individual veterinary practices are recommended.

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.

Sepsis is a life-threatening condition arising from a dysregulated host immune response to infection, leading to a substantial global health burden. The accurate identification of bacterial pathogens in sepsis is essential for guiding effective antimicrobial therapy and optimising patient outcomes. Traditional culture-based bacterial typing methods present inherent limitations, necessitating the exploration of alternative diagnostic approaches. This study reports the successful application of Fourier-transform infrared (FT-IR) spectroscopy in combination with chemometrics as a potent tool for the classification and discrimination of microbial species and strains, primarily sourced from individuals with invasive infections. These samples were obtained from various children with suspected sepsis infections with bacteria and fungi originating at different sites. We conducted a comprehensive analysis utilising 212 isolates from 14 distinct genera, comprising 202 bacterial and 10 fungal isolates. With the spectral analysis taking several weeks, we present the incorporation of quality control samples to mitigate potential variations that may arise between different sample plates, especially when dealing with a large sample size. The results demonstrated a remarkable consistency in clustering patterns among 14 genera when subjected to principal component analysis (PCA). Particularly, Candida, a fungal genus, was distinctly recovered away from bacterial samples. Principal component discriminant function analysis (PC-DFA) allowed for distinct discrimination between different bacterial groups, particularly Gram-negative and Gram-positive bacteria. Clear differentiation was also observed between coagulase-negative staphylococci (CNS) and Staphylococcus aureus isolates, while methicillin-resistant S. aureus (MRSA) was also separated from methicillin-susceptible S. aureus (MSSA) isolates. Furthermore, highly accurate discrimination was achieved between Enterococcus and vancomycin-resistant enterococci isolates with 98.4% accuracy using partial least squares-discriminant analysis. The study also demonstrates the specificity of FT-IR, as it effectively discriminates between individual isolates of Streptococcus and Candida at their respective species levels. The findings of this study establish a strong groundwork for the broader implementation of FT-IR and chemometrics in clinical and microbiological applications. The potential of these techniques for enhanced microbial classification holds significant promise in the diagnosis and management of invasive bacterial infections, thereby contributing to improved patient outcomes.

The global antimicrobial resistance crisis has been the driver of several international strategies on antimicrobial stewardship. For their implementation at the field level, the veterinary sector encounters several specific challenges and in particular: (i) a shortage of experts in key disciplines related to antimicrobial stewardship, (ii) a lack of evidence-based antimicrobial treatment guidelines, and (iii) inferior diagnostic tests available compared to human medicine. The present white paper describes how the COST Action ENOVAT (the European Network for Optimization of Veterinary Antimicrobial Treatment, CA18217), comprising 332 persons from 51 countries, worked towards solutions to these challenges. Initially, surveys were conducted to explore the present state in Europe in terms of existing antimicrobial use guidelines and microbiology practices performed. Concurrently, various research activities were launched to optimize diagnostics, including development of epidemiological cut-offs, clinical breakpoints and matrix-assisted laser desorption ionization time of flight mass spectrometry interpretive criteria. Also, guidelines drafting groups working towards evidence-based antimicrobial treatment guidelines for six conditions in food-producing and companion animals were established. The processes and outcomes, also in terms of capacity building, are summarized in this white paper where emphasis is placed on sustainability of the activities. Although several ENOVAT initiatives and spin-off projects will continue beyond the Action, we recommend that a new European veterinary research agenda is launched focusing on research and funding leading to long-term impacts on veterinary antimicrobial use.

Introduction Veterinary clinical microbiology laboratories play a key role in antimicrobial stewardship, surveillance of antimicrobial resistance and prevention of healthcare associated-infections. However, there is a shortage of international harmonized guidelines covering all steps of veterinary bacterial culture from sample receipt to reporting.Methods In order to gain insights, the European Network for Optimization of Veterinary Antimicrobial Treatment (ENOVAT) designed an online survey focused on the practices and interpretive criteria used for bacterial culture and identification (C&ID), and antimicrobial susceptibility testing (AST) of animal bacterial pathogens.Results A total of 241 microbiology laboratories in 34 European countries completed the survey, representing a mixture of academic (37.6%), governmental (27.4%), and private (26.5%) laboratories. The C&ID turnaround varied from 1 to 2 days (77.8%) to 3-5 days (20%), and 6- 8 days (1.6%), with similar timeframes for AST. Individual biochemical tests and analytical profile index (API) biochemical test kits or similar were the most frequent tools used for bacterial identification (77% and 56.2%, respectively), followed by PCR (46.6%) and MALDI-TOF MS (43.3%). For AST, Kirby-Bauer disk diffusion (DD) and minimum inhibitory concentration (MIC) determination were conducted by 43.8% and 32.6% of laboratories, respectively, with a combination of EUCAST and CLSI clinical breakpoints (CBPs) preferred for interpretation of the DD (41.2%) and MIC (47.6%) results. In the absence of specific CBPs, laboratories used human CBPs (53.3%) or veterinary CBPs representing another body site, organism or animal species (51.5%). Importantly, most laboratories (47.9%) only report the qualitative interpretation of the result (S, R, and I). As regards testing for AMR mechanisms, 48.5% and 46.7% of laboratories routinely screened isolates for methicillin resistance and ESBL production, respectively. Notably, selective reporting of AST results (i.e. excluding highest priority critically important antimicrobials from AST reports) was adopted by 39.5% of laboratories despite a similar proportion not taking any approach (37.6%) to guide clinicians towards narrower-spectrum or first-line antibiotics.Discussion In conclusion, we identified a broad variety of methodologies and interpretative criteria used for C&ID and AST in European veterinary microbiological diagnostic laboratories. The observed gaps in veterinary microbiology practices emphasize a need to improve and harmonize professional training, innovation, bacterial culture methods and interpretation, AMR surveillance and reporting strategies.

Antimicrobial resistance is a major public health concern worldwide. This study aims to determine the prevalence of Enterobacterales producing beta-lactamase (TEM, SHV, OXA) or extended-spectrum beta-lactamases (ESBL), as well as plasmid-mediated resistance to quinolones (PMQR) (qnrA, qnrB, qnrS) in companion animals from the northeast region of Romania. A total of 124 faecal samples were collected aseptically from healthy dogs attending the veterinary practice for vaccination and cultivated on Brilliance ESBL medium (Oxoid, UK). The ESBL production testing was performed using the combination disc test. The identification of Enterobacterales strains was achieved using molecular identification and based on biochemical tests. Antimicrobial susceptibility testing was performed using the disk diffusion method. Identification of genes encoding for beta-lactamase enzymes and genes encoding plasmid-mediated resistance to quinolones was performed by PCR according to the protocols previously described. After ESBL screening, 31 (31/124; 25%) extended-spectrum cephalosporin (ESC)-resistant Enterobacterales were obtained, and 67.74% (21/31) of them were confirmed as ESBL-producers. Regarding the Enterobacterales species, 27 (27/31; 87.1%) were Escherichia coli and 4 (4/31; 12.9%) strains were Klebsiella pneumoniae. Among the ESBL-producing isolates, the blaCTX-M-1 gene group was predominant (58.82%), followed by the blaCTX-M-9 group (41.18%). The blaTEM, blaSHV and blaOXA gene groups were identified in 54.83%, 29.03% and 3.22% of the analysed strains, respectively. The prevalence of PMQR genes was 22.58% and consisted only of qnrS (19.35%) and qnrA (3.22%) genes. The prevalence of ESBL strains related to the total number of analysed samples was 16.93% (21/124). The findings show a significant prevalence of ESBLs and PMQR genes in Enterobacterales strains isolated from the faeces of healthy dogs, implying that pets may pose a risk of transmitting ESBL strains to other animals or owners.

BackgroundCarbapenem-resistant Acinetobacter baumannii is a common pathogen associated with healthcare-acquired infections, and robust infection prevention and control protocols exist in human healthcare settings. In contrast, infection prevention and control (IPC) standards are limited in veterinary medicine, necessitating further investigation.AimExamine the possible transmission of carbapenem-resistant Acinetobacter spp. in a veterinary practice where a cat was diagnosed with an OXA-23-producing A. baumannii ST2 strain.MethodsEnvironmental samples together with nasal and hand swabs from the veterinary personnel were collected. All swabs were screened for the presence of extended-spectrum-β-lactamase- and carbapenemase-producing Enterobacterales, meticillin-resistant staphylococcus and multi-drug-resistant Acinetobacter spp. Whole-genome sequencing was performed for carbapenemase-producing strains.ResultsOf the veterinary staff, 60% carried meticillin-resistant Staphylococcus epidermidis. Environmental evaluation showed that 40% (N=6/15) of the surfaces analysed by contact plates and 40% (N=8/20) by swabs failed the hygiene criteria. Assessment of the surfaces revealed contamination with five OXA-23-producing Acinetobacter spp. strains: an OXA-23-producing Acinetobacter schindleri on the weight scale in the waiting room; and four OXA-23-producing Acinetobacter lwoffii strains, on different surfaces of the treatment room. The blaOXA-23 gene was located on the same plasmid-carrying Tn2008 across the different Acinetobacter spp. strains. These plasmids closely resemble a previously described OXA-23-encoding plasmid from a human Portuguese nosocomial Acinetobacter pittii isolate. Distinctly, the OXA-23-producing A. baumannii ST2 clinical strain had the resistant gene located on Tn2006, possibly inserted on the chromosome.ConclusionThe detection of an OXA-23-producing A. baumannii ST2 veterinary clinical strain is of concern for companion animal health and infection, prevention and control. This study established the dynamic of transmission of the plasmid-mediated blaOXA-23 gene on critical surfaces of a small animal veterinary practice. The genetic resemblance to a plasmid found in human nosocomial settings suggests a potential One Health link.

IntroductionHealthcare-associated infections (HCAIs) associated with extended-spectrum cephalosporin-resistant gram-negative (ESC-R GN) bacteria are an emerging concern in veterinary hospitals, especially in companion animal intensive care units (ICUs).MethodsTo understand the molecular epidemiology of ESC-R GN isolates in two veterinary hospitals (equine and small animal), a 6-month pilot study was performed during which fecal and environmental samples were obtained twice from selected patients, upon ICU admission and after 48 h of hospitalization. In total, 295 ESC-R GNs were analyzed using the Acuitas Resistome® Test (OpGen, Maryland, US), a PCR-based assay screening for 50 antimicrobial resistance gene families encoding for production of extended-spectrum beta-lactamase (ESBLs), TEM/SHV/OXA or AmpC beta-lactamases and carbapenemases. Combining organism identification and antimicrobial susceptibility data to genotyping results, unique “Acuitas profiles” were generated that can be used for fast typing the isolates and tracking transmission events.ResultsESKAPE GN pathogens were the most prevalent ESC-R GN isolates circulating in both the small animal and equine hospitals, consisting of Enterobacter cloacae complex (21.7%), Pseudomonas aeruginosa (20%), Klebsiella pneumoniae (15.9%), and Acinetobacter baumannii complex (13.6%) followed by Escherichia coli (12.2%), most harboring a combination of genes encoding for beta-lactamases and ESBLs. Some ESKAPE genotypes showed likely intra-hospital transmission, including E. cloacae (two genotypes, one carrying SHV4, SHV5, and TEM7 and the other TEM1, TEM3, and TEM7 enzymes) in the equine and K. pneumoniae (SHV1, SHV5, and DHA1-positive) in the small animal ICUs, respectively. Furthermore, P. aeruginosa (carrying OXA-50), A. baumannii complex (OXA-51), and E. coli (CTX-M-1) genotypes were isolated across both hospitals, suggesting possible transfer mediated via movement of staff and students. Importantly, isolates carrying transmissible resistance to last-resort antimicrobials (i.e. carbapenems) were identified within the hospital environments, consisting of three environmental Acinetobacter spp. harboring blaOXA − 23 and one clinical E. coli with blaOXA − 48.ConclusionWe describe the widespread occurrence of ESKAPE gram-negative organisms in veterinary ICU patients and hospital environments. Findings from this project provide baseline data on the epidemiology of ESKAPE pathogens in veterinary settings, which can inform infection control policies to aid in patient management and prevent transmission of nosocomial infections associated with these pathogens.