Escherichia coli (E. coli) is one of the most common pathogens in both humans and livestock. This study aimed to investigate the prevalence of E. coli isolated from raw cow milk and evaluate its antimicrobial resistance rates. A total of 1696 milk samples were collected from Romanian dairy farms from 2018 to 2022. E. coli was isolated on various selective agar media, such as Cled agar and Columbia Agar with 5% Sheep Blood. The identification of E. coli was performed by MALDI-TOF MS. E. coli isolates were tested for their susceptibility against 18 commonly used antibiotics in a disk diffusion method. The overall prevalence of E. coli was 22.45% of all isolated pathogens. Antibiogram analysis revealed that 27.51% of E. coli isolates from milk were multidrug-resistant. Resistance was highest for penicillin–novobiocin (87.78%), followed by streptomycin (53.7%). Resistance to six drugs (amoxicillin, streptomycin, kanamycin–cephalexin, marbofloxacin, ampicillin) showed a significant increasing trend over time, while for two drugs (penicillin G-framycetin, doxycycline), a significant decrease was observed. Our results suggest that milk can be a reservoir of bacteria with the potential for infection in humans via the food chain. Furthermore, there is a need for surveillance and monitoring to control the increase in resistance to currently used antimicrobials in dairy farms because the occurrence of multidrug-resistant E. coli isolated from milk poses a health hazard to consumers.

Despite the conjunctival vaccination program against Brucellosis in Palestine for more than 15 years, brucellosis continues to be a big problem concern, with increased incidence widely noted. Brucella melitensis is a facultative intracellular bacterium that replicates within macrophages. Recent reports showed that certain cell envelope associated genes of the B. melitensis is associated with its environmental stress adaptation and intracellular survival. Identification and characterization of these genes are essential to implementing efficient disease control and prevention systems. The aim of this study is to identify the cell envelope virulence associated genes of B. melitensis strains in Palestine. A total of 80 non-repetitive B. melitensis isolates were obtained from milk samples collected from infected small ruminants in the northern districts of the West Bank, Palestine during 2013 and 2015. The milk were cultured on Brucella agar (Oxoid) then incubated at 37°C for 3 to7 days. Positive cultures were identified by standard biochemical methods and confirmed by IS711 and Rev 1 rpsL gene PCR. All samples were negative for vaccine strain Rev 1 rpsL gene-PCR. The 80 B. melitensis under investigation were tested by multiplex PCR for 19 cell envelope virulence associated genes. Interestingly, our results revealed that all the 80 isolates were positive for the following genes: lpsB, lpsA rfbD, wbpL, pgm omp19, omp25, omp10, mgtA, uppS, amiC and dacF, while the other genes: wbdA, wbpZ, pmm, wbkA, perA, manB and wpbW were detected in more than 95% of the isolates

Background: The variability of prion protein gene (PRNP) codons and the frequency of alleles (K222, D146, and S146) that appear to confer genetic resistance to classical scrapie are still unknown in several goat populations/breeds prevalent in Romania. This work aims to assess the genetic polymorphism at 15 PRNP codons in Romanian goat populations to inform the development of goat breeding programs for scrapie resistance. Methods: Whole blood and hair follicles from Carpathian (50), French Alpine (53), and Banat’s White (53) breed goats were sampled to extract genomic DNA for genetic analyses and Sanger sequencing. In the targeted goat groups, one classical scrapie-positive Banat’s White goat was included. Results: The codons without polymorphisms were G37G, W102W, N146N, R151R, S173S, and I218I. The following non-synonymous polymorphisms of PRNP were recorded: P110P, P110S, P110T, T110T, G127G, G127S, I142I, I142M, T142I, H143H, P143P, R143R, R154R, H154R, P168P, Q168Q, Q211Q, Q211R, Q222Q, H222Q, K222K, S240S, P240P, P240S, and S240P. Conclusions: PRNP polymorphism was recorded in 60% (9/15) of codons. The scrapie-positive Banat’s White goat had G37G, W102W, T110T, G127G, I142I, H143H, N146N, R151R, R154R, P168P, S173S, R211R, I218I, Q222Q, and S240S. The K222 allele had a frequency of 6% (3/50) in Carpathian, 9.43% (5/53) in Banat’s White, and 15.09% (8/53) in French Alpine. Therefore, the polymorphisms detected in this sample of Romanian goat breeds are too rare to design a breeding program at the current time.

The behavior of Listeria monocytogenes (L. monocytogenes) throughout the shelf life of ready-to-eat foodstuffs represents a major concern in relation to human diet and human health. The aim of the study was to evaluate the behavior of L. monocytogenes in Wiener sausage, as an RTE meat product, throughout 15 days of storage (0–7 °C) under the action of fermented juice from parsley (Petroselinum crispum var. tuberosum) roots and common hawthorn (Crataegus monogyna) berry phenolics, compared with the effect of the food additives sodium nitrite and sodium ascorbate used in the standard formulation. For this purpose, one experimental formulation (F1) and one standard formulation (F2) of Wiener sausages were designed using the following preservatives and antioxidants: 50 ppm fermented parsley root juice (as a nitrite source) and 50 ppm hawthorn berry phenolics were used in F1, and 50 ppm sodium nitrite (as food additive E 250) and 50 ppm sodium ascorbate (as food additive E 301) were used in F2. The ability to support L. monocytogenes growth was assessed by a challenge test throughout the 15 days of storage. Based on the results of the assessment, the natural ingredients fermented parsley root juice and hawthorn berry phenolics could act as preservatives that ensure microbiological safety during the shelf life of the product. The nitrite and phenolic compounds of these natural ingredients showed antimicrobial activity against foodborne pathogens, including L. monocytogenes.

Listeria monocytogenes is a ubiquitous Gram-positive bacterium responsible for listeriosis, a foodborne zoonotic disease affecting humans and animals. Although infection in immunocompetent individuals is often asymptomatic or limited to mild self-limiting gastroenteritis, Listeria monocytogenes may cause severe invasive disease in vulnerable groups, including pregnant women, neonates, elderly individuals, and immunocompromised patients. Although the incidence of listeriosis is relatively low compared with many other foodborne pathogens, the high hospitalization and mortality rates associated with clinical cases make this bacterium a major concern for food safety and public health. The evolutionary success of L. monocytogenes reflects the interaction between a conserved core genome and a dynamic accessory genome shaped by horizontal gene transfer (HGT), ecological selection, and expansion of specific clones. Transient intestinal carriage in humans and animals, potentially influenced by gut microbiome composition, creates ecological interfaces where plasmids, transposons, prophages, and integrative conjugative elements contribute to the exchange of antimicrobial resistance determinants, virulence factors, and stress tolerance systems. Virulence diversification is further influenced by the differential distribution of pathogenicity islands such as LIPI-1, LIPI-3, and LIPI-4 across specific clonal lineages. These evolutionary processes occur across interconnected farm, food-production, environmental, and clinical ecosystems consistent with the One Health framework. Advances in whole-genome sequencing have clarified lineage-specific gene flow, expansion of specific clones, and the dynamics of the resistome and mobilome in L. monocytogenes populations. This narrative review aims to synthesize current knowledge on the mobile genetic elements and ecological interfaces that shape horizontal gene transfer in L. monocytogenes. Its novelty lies in integrating antimicrobial resistance, virulence-associated genomic islands, stress adaptation, and gut microbiome-mediated selection within a One Health and metapopulation framework. The main message of this review is that HGT should be interpreted as a context-dependent contributor to L. monocytogenes adaptation, acting together with clonal background, ecological selection, and mobile genetic elements.

African swine fever (ASF) remains one of the most devastating transboundary diseases affecting domestic pigs and wild boars worldwide, making molecular epidemiology essential for understanding viral transmission and supporting disease control. This study aimed to characterise African swine fever virus (ASFV) isolates circulating in Romania through comparative analysis of the B646L (p72) gene and the B602L central variable region (CVR). Fifty-four ASFV-positive samples collected from domestic pigs and wild boars were confirmed by real-time PCR and subjected to molecular analysis. Following Sanger sequencing and sequence quality assessment, 32 reliable consensus sequences were retained for B602L and 33 for p72. Sequencing of the p72 gene demonstrated a very high degree of nucleotide conservation, confirming that all successfully sequenced isolates belonged to genotype II. In contrast, the B602L region exhibited limited intra-genotypic variability characterised by scattered single nucleotide polymorphisms and localised insertion/deletion events, providing additional molecular resolution compared to p72, although insufficient for fine-scale epidemiological tracking. No genetic segregation according to host species was observed for either genomic region, and sequences obtained from domestic pigs and wild boar were interspersed in the phylogenetic analyses, demonstrating a high degree of genetic similarity at the investigated loci. These findings highlight the complementary value of p72 and B602L for ASFV molecular characterisation, while also indicating that these partial genomic markers alone cannot resolve transmission pathways or epidemiological links between domestic and wild host populations.