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

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.

In November 2024, an outbreak of highly pathogenic avian influenza (HPAI) H5N1 was confirmed in backyard poultry in Timiș County, Western Romania. The index cases involved chickens and domestic geese found dead with lesions characteristic of HPAI. Laboratory confirmation was achieved by real-time RT-qPCR targeting the matrix, H5, and N1 genes, followed by virus isolation in embryonated specific-pathogen-free eggs. Sequencing of the hemagglutinin cleavage site revealed the multi-basic motif PLREKRRKR/GLFG, consistent with a highly pathogenic phenotype. To investigate potential viral persistence, tracheal and cloacal swabs were collected from apparently healthy selected backyard poultry (chickens, geese, ducks and pheasants). RNA extraction and RT-qPCR, performed using protocols validated by the European Union Reference Laboratory for Avian Influenza, yielded negative results for all samples. Internal controls confirmed assay reliability, excluding the possibility of PCR inhibition. The investigation confirmed the occurrence of HPAI H5N1 in backyard poultry and demonstrated the absence of detectable viral persistence in surrounding flocks under the tested conditions. These findings highlight the importance of rapid molecular diagnostics, active surveillance, and strict biosecurity in limiting virus spread. Continued monitoring under the One Health framework is essential to mitigate the risk of avian influenza at the human–animal–environment interface.

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) is one of the most severe contagious viral diseases affecting domestic pigs and wild boars. The African swine fever virus is the only member of the Asfarviridae family, it is the only DNA virus transmitted by arthropods in which it can persist for a period longer than 5 years. The disease was eradicated from Europe (except Sardinia) in the 1990s through drastic control and eradication programs. The introduction of the virus into Romania was caused by the unofficial introduction of pork products purchased from Ukraine. Romania reported to WOAH for the year 2021 a number of 2618 new outbreaks of ASF, of which 1199 in the first semester and 1419 in the second semester. Of these, 1628 were declared in the domestic pig population and 990 outbreaks in wild boars. In the first semester of 2022, Romania recorded 775 new outbreaks of ASF, of which 336 outbreaks were declared in the domestic pig population and 439 in wild boars. In the second half of 2022, 315 new outbreaks of ASF were declared, of which 167 outbreaks were recorded in domestic pigs and 148 in wild boars. In 2023, 819 new outbreaks of ASF were reported on the territory of our country. Of these, 530 outbreaks were identified in the domestic pig population and 289 in wild boars. In Romania's latest notification to WOAH, 507,000 cases in domestic pigs and 597 in wild boars are recorded. Thus, in Romania, in the period 2017-2024, a number of 7,450 new outbreaks of ASF were reported to WOAH. Considering the number of African swine fever outbreaks in our country in the last 7 years, and the endemicity of the infection, monitoring of pigs and wild boars, and very rigorous control of their movement are the only tools available to control the disease.

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.