Foodborne pathogenic bacteria, like Salmonella spp. and Listeria monocytogenes, can be detected in the primary food production environment. On the other hand, and in the current context of One Health, antimicrobial resistance (AMR) is gaining increased attention worldwide, as it poses significant threat to public health. The purpose of this study was to confirm the presence of Salmonella spp. and L. monocytogenes in pet food and feed samples, by means of biochemical and/or serological testing of the microbial isolates, and then to screen for AMR against a panel of selected antibiotics. Serotyping of the isolates with multiplex polymerase chain reaction revealed the presence of three of the most common clinical Salmonella serovars (S. Enteritidis, S. Typhimurium, S. Thompson) and the major epidemiologically important L. monocytogenes serotypes (1/2a, 1/2b, 1/2c, 4b) in 15 and 9 confirmed isolates of the pathogens, respectively. Strains of Salmonella spp. showed resistance to tetracycline (n = 3) and combined AMR to tetracycline with either ampicillin (n = 2) or trimethoprim-sulfamethoxazole (n = 3), without any multidrug resistance (MDR) being recorded whatsoever. AMR in L. monocytogenes was documented in 55.5% of the bacterial strains (n = 5) tested against ciprofloxacin, meropenem, penicillin, trimethoprim-sulfamethoxazole, and tetracycline. Alarmingly, one strain of L. monocytogenes was MDR to the latter five antibiotics and deemed resistant in three antibiotic groups (carbapenems, penicillins, tetracyclines), after exhibiting minimum inhibitory concentrations (MICs) to meropenem (MIC = 4 μg/mL), penicillin (MIC = 4 μg/mL), and tetracycline (MIC = 48 μg/mL). To the best of our knowledge, finding an MDR L. monocytogenes in pet food is something reported for the first time herein. The results presented in this study highlight the presence of important foodborne bacterial pathogens, such as Salmonella spp. and L. monocytogenes, with increased AMR to antibiotics and possible MDR at the primary production and at the farm level, due to the misuse of pharmacological substances used to treat zoonotic diseases, probably resulting in detection of resistant strains of these pathogenic bacteria in animal-originated food products (e.g., meat, milk, eggs).

Background/Objectives: Over the past decade, extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli have emerged as a significant public health concern due to their potential to spread beyond clinical settings and healthy carriers, in various environments, including in animal source foods. This study seeks to investigate the molecular characteristics of ESBL-producing E. coli strains isolated from food of animal origin, with a focus on chromosomal typing, plasmid typing, and the description of the associated resistance genes’ genetic environment. Methods: Ninety-seven food of animal origin samples were tested for E. coli isolates resistant to cefotaxime. The resulting isolates were then subjected to antimicrobial susceptibility testing and PCR analysis to detect beta-lactamase genes. Additional assays, encompassing mating-out procedures, molecular typing utilizing Pulsed-Field Gel Electrophoresis, Multilocus Sequence Typing Analysis, and Oxford Nanopore Technology Lite whole plasmid sequencing, were also conducted. Results: E. coli was detected in 26 raw food specimens, generating a percentage of 27%. Fourteen of the current isolates (14%) were resistant to third generation cephalosporins, producing CTX-M-1, CTX-M-15, CTX-M-55, and SHV-12 beta-lactamases. The respective genes were accompanied by Insertion Sequences ISEcp1 and IS26, facilitating their transfer. Among plasmids harboring ESBL genes, representatives belonging to incI1 incompatibility group prevailed (5/8), followed by IncY and IncX3. Most plasmids proved conjugative. Diversity of molecular fingerprints of ESBL producing E. coli was revealed. Conclusions: To the best our knowledge, this study is the first to describe the molecular characteristics of E. coli isolates producing ESBLs sourced from foods of animal origin in Greece. The prevalence of ESBLs in our confined food collection is primarily associated with the very successful IncI1 plasmids, which were not linked to a specific E. coli genetic background. This lack of association confirms that horizontal plasmid transfer plays a more significant role than clonal dissemination in the spread of ESBL-mediated cephalosporin resistance.

Listeria monocytogenes is already a well-known foodborne bacterial pathogen, ubiquitously dispersed not only in the food production environment but also in the primary animal production environment as well. The present study performed whole-genome characterization of the multidrug-resistant (MDR) L. monocytogenes strain BF11, previously isolated from raw pet food and phenotypically described for antimicrobial resistance. To this end, the genomic analysis performed on the isolate confirmed the pathogen’s designation as a serotype 1/2b strain belonging to ST5 and CC5 (Lineage I), carrying multiple MDR genes, stress-related genes, and mobile genetic elements, despite the absence of plasmids. The strain is phylogenetically closely related to Lineage I epidemic strains (e.g., F2365), as it has a full-length inlA and a functional prfA, rendering it capable of invading human cells and marking its high virulence. Overall, this strain may represent a potentially novel genomic profile when core genome multilocus sequence typing (cgMLST) is used, although further data from additional isolates would be required to confirm its classification within a new Complex Type, while displaying a hybrid unique profile. It is an evolved ST5 L. monocytogenes strain that has acquired genetic material conferring a “clinical signature” (Lineage I-like) and an extensive resistance network. Therefore, presence of L. monocytogenes strain BF11 in pet food is alarming, since such hybrid strains often evade surveillance monitoring as they do not fit strictly into classical categories, posing a serious food safety and public health threat in the concept of One Health.