Genomic and clinical case characterisation of Staphylococcus haemolyticus isolated from dogs and cats in the United States, including strains with high-level mupirocin tolerance.

BACKGROUND: Staphylococcus haemolyticus is a coagulase-negative commensal organism of both people and companion animals. It has pathogenic potential and when cultured is often meticillin- and multidrug-resistant. OBJECTIVES: To characterise the clinical features of dogs and cats with clinical skin disease that had positive S. haemolyticus skin cultures, and to employ whole-genome sequencing (WGS) to identify resistance genes and characterise the genetic relatedness of strains. MATERIALS AND METHODS: Isolates were identified by the institutional clinical microbiology laboratory by routine aerobic culture and susceptibility from seven veterinary hospitals across the United States. Then, WGS and analysis of each isolate were performed and clinical data collected via a retrospective clinician questionnaire. RESULTS: S. haemolyticus was identified from superficial (seven of 12) and deep (five of 12) cutaneous infections in our study. Most animals had received antimicrobials (10 of 12) and/or immunomodulatory drugs (nine of 12) within the six months before culture. WGS analysis revealed a variety of genetic lineages and a wide array of antimicrobial resistance genes. Meticillin resistance was identified in nine of 12 isolates and four of 12 isolates demonstrated mupirocin tolerance. CONCLUSIONS AND CLINICAL RELEVANCE: Staphylococcus haemolyticus may be an under-recognised pathogen in companion animals, and its demonstrated potential for multidrug-resistance, meticillin-resistance, and high-level mupirocin tolerance may create a therapeutic challenge. Further studies should evaluate the prior antimicrobial use and immunocompromised status as risk factors for infection with S. haemolyticus.

Use of a chromogenic medium with and without selective enrichment to screen for carbapenemase-producing Enterobacterales (CPE) from canine and feline fecal specimens during an outbreak of NDM-5-producing Escherichia coli.

Carbapenemase-producing Enterobacterales (CPE) are one of the most urgent threats to human healthcare globally. Descriptions of CPE outbreaks in veterinary hospitals suggest the need for screening strategies for CPE from companion animals. Our aim was to optimize a chromogenic agar method with and without selective enrichment to isolate CPE from companion animal feces in an ongoing outbreak of New Delhi metallo-ß-lactamse-5 Escherichia coli. A limit of detection (LOD) assay for spiked canine and feline feces was performed for both methods using a carbapenamase-producing E. coli (24213-18); the LOD (1.5 × 103 cfu/g of feces) was equivalent to that reported for human fecal specimens. We screened 1,247 companion animal fecal specimens for carriage of CPE by 1) direct plating to chromogenic agar and 2) plating to chromogenic agar following selective enrichment. Twenty-one specimens were positive for CPE by both direct culture and enrichment culture. No specimens were positive with selective enrichment and negative by direct culture. A selective enrichment step did not result in any increased recovery of CPE from companion animals, which suggests that enrichment broth may not be necessary for outbreak surveillance testing. It is important to continue to validate methods for the detection of CPE in companion animals as outbreaks become more common in veterinary facilities.

Prevalence and molecular epidemiology of carbapenemase-producing Enterobacterales isolated from dog and cat faeces submitted to veterinary laboratories in the USA.

AIMS: To estimate the prevalence of carbapenemase-producing Enterobacterales (CPE) carriage among pets using faecal specimens submitted to veterinary diagnostic laboratories throughout the US. A secondary aim was to employ whole-genome sequencing (WGS) to characterize isolates of CPE from companion animals and compare them to publicly available CPE genomes. METHODS AND RESULTS: To estimate the prevalence of CPE in companion animals in the USA, a multicenter surveillance study including 8 different veterinary diagnostic laboratories from across the USA was conducted. Briefly, remnant faecal specimens from dogs and cats were screened using two selective agar plates (CHROMID Carba and MacConkey with 1 mg/L cefotaxime and 0.125 mg/L meropenem) and presumptive CPE isolates screened by the modified carbapenemase inactivation method for carbapenemase production. A total of 2393 specimens were screened and yielded 196 isolates for carbapenemase screening. A total of 5 isolates from 4 dogs and 1 cat at 3 different veterinary diagnostic laboratories were confirmed to produce a carbapenemase (0.21%). Whole-genome sequencing (WGS) revealed two E. coli (ST167) isolates that both produced an NDM-5 carbapenemase, two Enterobacter hormaechei (ST171) isolates that produced an NDM-5 carbapenemase and a KPC-4 carbapenemase respectively and one Klebsiella oxytoca (ST199) that produced an Oxa-48-type carbapenemase. Both E. coli isolates were found to be within at least 22 SNPs of previously characterized canine and human CPE isolates. CONCLUSIONS: This study demonstrates that the prevalence of CPE among companion animals is relatively low (0.21%) but that given the genetic relatedness of animal isolates to human isolates, additional surveillance is needed.

Evaluation of canine raw food products for the presence of extended-spectrum beta-lactamase- and carbapenemase-producing bacteria of the order Enterobacterales.

OBJECTIVE: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity. SAMPLES: A total of 200 canine raw food products. METHODS: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order. RESULTS: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%). CLINICAL RELEVANCE: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.

Comparison of a Chromogenic Urine Culture Plate System (UTid+) and Conventional Urine Culture for Canine and Feline Specimens.

In companion animal medicine, urinary tract infection (UTI) is one of the most common indications for antimicrobial therapy. Definitive diagnosis of UTI requires isolation of bacteria with routine urine culture from an animal with concurrent clinical signs. Urine culture is typically performed at reference laboratories where paired susceptibility testing can be performed, but delays in shipment or processing can affect results. This study evaluated the use of a selective chromogenic, point-of-care culture system (UTid+) compared to conventional urine culture. A total of 119 (73 canine and 46 feline) cystocentesis urine samples were evaluated. Conventional urine culture was positive for 28 (23.5%) of the 119 cultures and UTid+ culture was positive for 26 (21.8%). The overall sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 92.3%, 97.8%, 92.3%, 97.8 and 96.6% for UTid+ respectively. Overall, the UTid+ culture system showed an acceptable level of accuracy when compared to conventional urine culture. Agreement of identification results was high (κ = 0.90) with an important exception being Proteus spp. which was only identified in 1/3 positive cultures. UTid+ may be useful in scenarios where a common UTI pathogen is expected and identification within 24 h is ideal; however, conventional urine culture remains the gold standard.