Listeria monocytogenes (L. monocytogenes) poses a significant threat to food safety due to its ability to cause severe human illness and its resistance to various antibiotics and environmental conditions. This study investigated the prevalence, serotype distribution, virulence gene profiles, and antimicrobial resistance patterns of L. monocytogenes in ready-to-eat (RTE) food products from Romania. A total of 8151 samples were analyzed, including various processed dairy, bovine, poultry, pork, and fish products. Bacterial isolation was conducted using the classical standard method, followed by confirmation through biochemical and molecular testing. Among the isolated strains, serotypes 1/2a, 1/2b, and 1/2c were identified, with a prevalence of 75% for serotype 1/2a. Additionally, virulence genes specific to listeriolysin O (hlyA) and regulatory factor A (prfA) were detected in all isolates. Antimicrobial susceptibility testing revealed varying resistance patterns among the L. monocytogenes strains. Trimethoprim-sulfamethoxazole and oxacillin showed the highest prevalence of resistance at 26.92% and 23.07%, respectively. However, all strains remained susceptible to ciprofloxacin, levofloxacin, and moxifloxacin. Notably, 23.07% of the isolates exhibited multidrug resistance, with the most common pattern being resistance to oxacillin, penicillin, and tetracycline. Analysis of antimicrobial resistance genes identified tetracycline resistance genes, particularly tet(C), tet(M), and tet(K), in a significant proportion of isolates. The presence of ampC and dfrD genes was also notable, indicating potential mechanisms of resistance. These results emphasize the necessity for ongoing surveillance of L. monocytogenes in RTE foods and emphasize the importance of thorough monitoring of antimicrobial resistance to guide public health strategies within the European Union.

Antimicrobial resistance has become a worldwide concern in all public health domains and reducing the spread has become a global priority. Pathogenic E. coli is responsible for a number of illnesses in humans and outbreaks in the past have been correlated with the consumption of contaminated bovine products. This is why surveillance in all the steps of production is essential. This study focused on identifying the pathogenic strains of E. coli in two large bovine abattoirs from Romania and France, and on associating them with the antimicrobial resistance patterns. A total of 250 samples from intestinal content were aseptically collected during the evisceration step of the cattle slaughtering process, from which 242 E. coli strains were isolated. Seventeen percent of all samples tested positive to at least one E. coli isolate carrying eaeA, stx1 and stx2 genes. The most prevalent genetic profile found in the E. coli strains tested was Stx1-positive and Stx2/eaeA-negative. More than 68% of the pathogenic E. coli isolated in Romania showed multi-drug resistance (MDR) and in France, the percentage was significantly lower (38%). The MDR profiles showed a high gene diversity for antibiotic resistance, which represents a great risk for environmental spread and human health. Our results indicate that in Romania, bovines can represent a reservoir for MDR E. coli and, hence, a surveillance system for antimicrobials usage in farm animals is highly needed.

Antibiotic resistance among pathogenic bacteria in humans and animals poses a critical public health challenge, leading to diminished effectiveness of existing antimicrobial treatments. Notably, animal-derived food products are significant vectors for the transmission of resistant bacteria to humans, with Salmonella species being predominant culprits in foodborne illnesses. This study investigates the prevalence and antibiotic resistance patterns of Salmonella serovars isolated from traditionally sourced pork meat in Romania. Over a four-year period, 208 pork samples were collected using standardized protocols during traditional slaughtering practices. Salmonella spp. were isolated following ISO 6579:2002 guidelines and confirmed using biochemical assays and PCR. Serotyping was performed using specific antisera, and antimicrobial susceptibility testing was conducted through the standard disk diffusion method, assessing 11 antibiotics. Results indicated a 23.07% prevalence of Salmonella, with 48 isolates categorized into eight serovars, primarily S. Infantis (n = 15), S. Typhimurium (n = 15), and S. Derby (n = 11). PCR results confirmed the presence of Salmonella by detecting the hilA and ompC genes, with 31.25% of isolates being positive for the Typhimurium-specific sequence. Notably, 93.75% of the isolates were multidrug-resistant (MDR), exhibiting high resistance rates against streptomycin (91.66%) (>10 µg), tetracycline (83.33%) (>30 µg), and sulfamethoxazole (68.75%) (>300 µg). More than 60% of MDR isolates displayed resistance to five or more antibiotics. These findings underscore the need for coordinated control measures in the pork production chain to combat the spread of Salmonella and protect public health. Enhanced surveillance and intervention strategies are crucial for addressing antibiotic resistance and reducing the risk of foodborne illnesses linked to contaminated animal products.

Food safety, particularly within the meat industry, is a significant concern addressed under the One Health concept, emphasizing the necessity of enhanced surveillance and hygiene protocols to mitigate contamination risks. This study assessed microbiological risks in Romanian bovine slaughterhouses by analyzing 150 samples from stool and carcasses at the post-evisceration and cooling stages over seven months in two abattoirs, using standardized microbiological methods and PCR to quantify aerobic colony counts (ACCs), Enterobacteriaceae, and pathogens (E. coli, Salmonella spp., and Listeria spp.). ACCs and Enterobacteriaceae levels decreased significantly [p < 0.05] during processing, highlighting effective hygiene measures. Pathogenic E. coli was identified in 14% of fecal samples and 5% of carcasses, indicating cross-contamination risks. Salmonella spp. were found in 28% of fecal samples but absent on carcasses, suggesting successful containment. Listeria spp. were rare and not detected on carcasses. PCR confirmed the presence of pathogenic strains in stool samples, emphasizing the need for strict hygiene practices and regular monitoring to improve meat safety and protect public health. In conclusion, the prevalence of E. coli, particularly serogroups like O101 and O26, and the absence of Salmonella and Listeria in carcass samples reflect both regional differences in pathogenic strains and the need for comprehensive, multi-stage control measures. Further studies should broaden pathogen surveillance to include more E. coli serogroups and implement stricter hygiene protocols to prevent cross-contamination during evisceration, skinning, and cooling. Regular monitoring of Salmonella and Listeria, especially in silage-fed cattle regions, along with improved coordination across the food production, health, and environmental sectors, is essential to mitigate contamination risks and safeguard public health.

The aim of this study was to determine the polycyclic aromatic hydrocarbons (PAH) levels in the smoked meat products obtained in the traditional system in Romania. The importance of this study resides in the fact that these compounds are known to be carcinogenic, and their levels should be strictly monitored to ensure the safety of the consumers. Until now, no study was performed in Romania that addresses this issue in the traditional pork meat products. The study was performed on 60 meat products samples (sausages, bacon, ham) taken from a certified traditional meat processing unit but also from retail units that delivered industrially processed meat products. The method used in the detection of the 15 PAHs analysed was HPLC. Our results showed that the highest percent of total PAHs is found in the traditionally obtained smoked bacon and the lowest in industrially produced pork meat sausage. None of the samples examined exceeded the limit imposed by the European legislation for benzo[a]pyrene (BaP), but the smoked bacon revealed higher values for PAH4 than the allowed limit. Following this study, we suggest that a more careful attention should be paid to all factors and apply such smoking conditions that result in the lowest possible contamination with PAH in pork meat products obtained in the traditional system in Romania.

Traditional pork meat products produced through artisanal smoking and drying techniques are highly appreciated for their distinctive sensory characteristics; however, such practices may raise concerns regarding both chemical and microbiological safety. The present study aimed to assess the occurrence of selected polycyclic aromatic hydrocarbons (PAHs) and hygiene- and safety-related microorganisms in traditionally processed pork meat products collected from local markets and small-scale producers. A total of 140 samples were analyzed for four marker PAHs—benzo[a]pyrene (BaP), benz[a]anthracene (BaA), benzo[b]fluoranthene (BbF), and chrysene (Chr)—using gas chromatography–mass spectrometry (GC–MS). Microbiological contamination was evaluated through standard plate count techniques, and the presence of Listeria monocytogenes and Salmonella serovars was determined using selective isolation methods, followed by PCR confirmation of pathogenic strains. PAH concentrations varied widely: BaP (0.3–1.8 µg/kg), BaA (0.5–2.4 µg/kg), BbF (0.8–3.1 µg/kg) and Chr (0.4–2.0 µg/kg), with ΣPAH₄ (Sum of PAH4, referring to the total concentration of the four-priority polycyclic aromatic hydrocarbons) ranging from 2.5 to 8.3 µg/kg. Smoked sausages showed the highest contamination (BaP: 1.8 µg/kg; ΣPAH₄: 8.3 µg/kg), significantly exceeding levels in dry-cured ham (BaP: 1.2 µg/kg; ΣPAH₄: 6.1 µg/kg) and smoked bacon (BaP: 0.9 µg/kg; ΣPAH₄: 5.4 µg/kg) (Kruskal–Wallis, p < 0.0001). Although all samples complied with the EU ΣPAH₄ limit (12 µg/kg), 15% exceeded the BaP limit of 2.0 µg/kg, primarily among artisanal sausages. Microbiological analyses revealed total coliform counts between 1.5 × 10² and 6.2 × 10⁴ CFU/g, while Enterobacteriaceae ranged from 2.0 × 10² to 4.9 × 10⁴ CFU/g. Samples obtained from unregulated producers exhibited higher bacterial loads, indicating suboptimal hygiene during processing and storage. A moderate positive correlation was identified between total coliform and Enterobacteriaceae counts (r = 0.59, p < 0.05). Moreover, Salmonella serovars was detected in ten sausage samples, and Listeria monocytogenes was confirmed in three samples of traditional products. Overall, the findings suggest that although PAH contamination generally complied with EU safety limits, occasional exceedances of benzo[a]pyrene and elevated microbial indicators underscore the need for stricter control of smoking parameters, fuel sources, and hygienic handling. Implementation of standardized smoking protocols and good manufacturing practices (GMP) is recommended to enhance the safety and quality of traditional pork meat products