Campylobacter remains the most prevalent foodborne pathogen bacterium responsible for causing gastroenteritis worldwide. Specifically, this pathogen colonises a ubiquitous range of environments, from poultry, companion pets and livestock animals to humans. The bacterium is uniquely adaptable to various niches, leading to complicated gastroenteritis and, in some cases, difficult to treat due to elevated resistance to certain antibiotics. This increased resistance is currently detected via genomic, clinical or epidemiological studies, with the results highlighting worrying multi-drug resistant (MDR) profiles in many food and clinical isolates. The Campylobacter genome encodes a rich inventory of virulence factors offering the bacterium the ability to influence host immune defences, survive antimicrobials, form biofilms and ultimately boost its infection-inducing potential. The virulence traits responsible for inducing clinical signs are not sufficiently defined because several populations have ample virulence genes with physiological functions that reflect their pathogenicity differences as well as a complement of antimicrobial resistance (AMR) systems. Therefore, exhaustive knowledge of the virulence factors associated with Campylobacter is crucial for collecting molecular insights into the infectivity processes, which could pave the way for new therapeutical targets to combat and control the infection and mitigate the spread of MDR bacteria. This review provides an overview of the spread and prevalence of genetic determinants associated with virulence and antibiotic resistance from studies performed on livestock animals. In addition, we have investigated the relevant coincidental associations between the prevalence of the genes responsible for pathogenic virulence, horizontal gene transfer (HGT) and transmissibility of highly pathogenic Campylobacter strains.

Campylobacter species, predominantly Campylobacter jejuni, remains a significant zoonotic pathogen worldwide, with the poultry sector being the primary vector for human transmission. In recent years. there has been a notable rise in the incidence of human campylobacteriosis, necessitating a deeper understanding of the pathogen's survival mechanisms and transmission dynamics. Biofilm presence significantly contributes to C. jejuni persistence in poultry and subsequent food product contamination, and this review describes the intricate processes involved in biofilm formation. The ability of Campylobacter to form biofilms on various surfaces, including stainless steel, plastic, and glass, is a critical survival strategy. Campylobacter biofilms, with their remarkable resilience, protect the pathogen from environmental stresses such as desiccation, pH extremes, biocides and sanitizing agents. This review explores the molecular and genetic mechanisms of C. jejuni biofilm formation, highlighting regulatory genes involved in motility, chemotaxis, and stress responses. Flagellar proteins, particularly flaA, flaB, flaG, and adhesins like cadF and flpA, are identified as the main molecular components in biofilm development. The role of mixed-species biofilms, where C. jejuni integrates into existing biofilms of other bacteria to enhance pathogen resilience, is also discussed. This review also considers alternative interventions to control C. jejuni in poultry production, in the context of increasing antibiotic resistance. It explores the effectiveness of prebiotics, probiotics, synbiotics, bacteriocins, bacteriophages, vaccines, and organic acids, with a focus on their mechanisms of action in reducing bacterial colonization and biofilm formation. Studies show that mixtures of organic acids and compounds like Carvacrol and Eugenol significantly downregulate genes linked with motility and adhesion, thereby disrupting biofilm integrity. It discusses the impact of environmental factors, such as temperature and oxygen levels on biofilm formation, providing insights into how industrial conditions can be manipulated to reduce contamination. This paper stresses the need for a multifaceted approach to control Campylobacter in poultry, integrating molecular and genetic insights with practical interventions. By advancing our understanding of biofilm dynamics and gene regulation, we aim to inform the development of more effective strategies to enhance food safety and protect public health.

Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis can colonize the tooth root canals, adhere to dentin walls, and frequently cause periodontitis in dogs. Bacterial periodontal diseases are common in domesticated pets, causing severe oral cavity inflammation and a strong immune response. This study investigates the antioxidant effect of a natural antimicrobial mixture (Auraguard—Ag) on the ability of S. aureus, S. pyogenes and E. faecalis to infect primary canine oral epithelial cells as well as its impact on their virulence factors. Our data show that a concentration of 0.25% Ag is sufficient to inhibit the growth of all three pathogens, whereas a concentration of 0.5% will become bactericidal. The sub-inhibitory concentration of 0.125% Ag reveals that the antimicrobial mixture can significantly reduce biofilm formation and exopolysaccharide production. The impact on these virulence factors was further translated into a significantly reduced ability to infect primary canine oral epithelial cells and restore epithelial tight junctions, with no impact on the epithelial cell viability. The post-infection inflammatory cytokines (IL-1β and IL-8) and the COX-2 mediator were also reduced both in mRNA and protein expression levels. The oxidative burst, detected upon infection, was also decreased in the presence of Ag, as our results show a significant decrease in H₂O₂ released by the infected cells. We show that inhibition of either NADPH or ERK activity will result in a downregulation of COX-2 expression and lower levels of H₂O₂ in infected cells. Conclusively, our study shows that natural antimicrobials reduce pro-inflammatory events, post infection, through an antioxidative mechanism that involves the downregulation of the COX-2 mediator via the inactivation of ERK in the absence of H₂O₂. As a result, they significantly reduce the risk of secondary bacterial infections and host oxidative stress caused by Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis accumulation in biofilms in an in vitro canine oral infection model.

The objective of this work was to investigate, for the first time, the antioxidant effect of a mixture of natural antimicrobials in an Enterocytozoon hepatopenaei (EHP) shrimp-gut model of infection and the biological mechanisms involved in their way of action. The study approach included investigations, firstly, in vitro, on shrimp-gut primary (SGP) epithelial cells and in vivo by using EHP-challenged shrimp. Our results show that exposure of EHP spores to 0.1%, 0.5%, 1%, and 2% AuraAqua (Aq) significantly reduced spore activity at all concentrations but was more pronounced after exposure to 0.5% Aq. The Aq was able to reduce EHP infection of SGP cells regardless of cells being pretreated or cocultured during infection with Aq. The survivability of SGP cells infected with EHP spores was significantly increased in both scenarios; however, a more noticeable effect was observed when the infected cells were pre-exposed to Aq. Our data show that infection of SGP cells by EHP activates the host NADPH oxidases and the release of H₂O₂ produced. When Aq was used during infection, a significant reduction in H₂O₂ was observed concomitant with a significant increase in the levels of CAT and SOD enzymes. Moreover, in the presence of 0.5% Aq, the overproduction of CAT and SOD was correlated with the inactivation of the NF-κB pathway, which, otherwise, as we show, is activated upon EHP infection of SGP cells. In a challenge test, Aq was able to significantly reduce mortality in EHP-infected shrimp and increase the levels of CAT and SOD in the gut tissue. Conclusively, these results show, for the first time, that a mixture of natural antimicrobials (Aq) can reduce the EHP-spore activity, improve the survival rates of primary gut-shrimp epithelial cells and reduce the oxidative damage caused by EHP infection. Moreover, we show that Aq was able to stop the H₂O₂ activation of the NF-κB pathway of Crustins, Penaeidins, and the lysozyme, and the CAT and SOD activity both in vitro and in a shrimp challenge test.