Enterovirus E (EV-E), a representative of the Picornaviridae family, endemically affects cattle across the world, typically causing subclinical infections. However, under favorable conditions, severe or fatal disorders of the respiratory, digestive, and reproductive systems may develop. There is no specific treatment for enterovirus infections in humans or animals, and only symptomatic treatment is available. The aim of this study was to determine the in vitro antiviral effect of bovine lactoferrin (bLF) against enterovirus E using virucidal, cytopathic effect inhibition, and viral yield reduction assays in MDBK cells. The influence of lactoferrin on the intracellular viral RNA level was also determined. Surprisingly, lactoferrin did not have a protective effect on cells, although it inhibited the replication of the virus during the adsorption and post-adsorption stages (viral titres reduced by 1–1.1 log). Additionally, a decrease in the viral RNA level in cells (by up to 75%) was observed. More detailed studies are needed to determine the mechanism of bovine lactoferrin effect on enterovirus E. However, this highly biocompatible protein ensures some degree of protection against infection by bovine enterovirus, which is particularly important for young animals that receive this protein in their mother’s milk.

Background: Pseudomonas aeruginosa is a pathogen that causes infections in animals and humans, with veterinary implications including ear infections in dogs, respiratory diseases in cats, and mastitis in ruminants. In humans, it causes severe hospital-acquired infections, particularly in immunosuppressed patients. This study aimed to identify and assess the prevalence of specific virulence factors in Pseudomonas aeruginosa isolates. Methods: We analyzed 98 Pseudomonas aeruginosa isolates from various animal samples (dogs, cats, ruminants, fowl) from northeastern Poland in 2019–2022 for virulence-related genes (toxA, exoU, exoT, exoS, lasB, plcN, plcH, pldA, aprA, gacA, algD, pelA, endA, and oprF) by PCR and assessed biofilm formation at 48 and 72 h. Genomic diversity was assessed by ERIC-PCR. Results: The obtained results showed that all strains harbored the pelA gene (100%), while the lowest prevalence was found for pldA (24%) and exoU (36%). Regardless of the animal species, strong biofilm forming ability was prevalent among the strains after both 48 h (75%) and 72 h (74%). We obtained as many as 87 different genotyping profiles, where the dominant one was profile ERIC-48, observed in four strains. Conclusions: No correlation was found between presence or absence of determined genes and the nature of infection. Similarly, no correlation was found between biofilm-forming genes and biofilm strength. The high genetic diversity indicates challenges for effective prevention, emphasizing the need for ongoing monitoring and research.

Enterovirus eibovi (EV-E) is a bovine RNA virus with debated pathogenic potential. The aim of this study was to evaluate the in vitro effects of bovine lactoferrin (bLF), an antimicrobial and immunomodulatory glycoprotein, on EV-E-infected bovine peripheral blood mononuclear cells (PBMC). The PBMC viability, phagocytic function, and lymphocyte proliferation were examined using spectrophotometric assays. Flow cytometry was used for immunophenotyping of lymphocytes, and real-time quantitative PCR for evaluation of immune-related gene expression. The safe concentration range of bLF (1-25 µg/mL) was determined on the basis of the viability and proliferation of uninfected PBMC. Bovine lactoferrin significantly influences the activity of enterovirus-infected cells, with high (10 and 25 µg/mL) and low (1, 2.5, and 5 µg/mL) protein concentrations exerting different effects. Low concentrations of bLF primarily affected unstimulated cells, increasing the expression of IFNα, IFNβ, IL2, and NFκB genes. In contrast, high concentrations of lactoferrin predominantly modulated the activity of infected cells that were additionally stimulated with a mitogen or antigen: They increased T lymphocyte proliferation; reduced phagocyte activity; increased the expression of IFNα, IL10, and NFκB; and decreased the expression of IFNγ. The increased expression of IL10 in response to high protein concentrations was also observed in unstimulated cells. The only common effect of both high and low concentrations of bLF was a decrease in the percentage of WC1+ γδ T cells following concanavalin A stimulation. Thus, bLF modulated the immune response of EV-E-infected PBMC in a concentration-dependent manner: Lower concentrations promoted early antiviral responses by inducing type I interferons and IL-2, whereas higher concentrations upregulated the anti-inflammatory cytokine IL-10, suggesting a protective effect against immune overstimulation. These findings indicate that bLF may serve as an immunomodulatory agent during enteroviral infections in cattle, with promising implications for both therapeutic and prophylactic applications.

Introduction Enterovirus E (EV-E) is a common viral pathogen endemic in cattle worldwide. Little is known, however, about its potential interactions with bovine immune cells. Material and Methods The EV-E-permissiveness of bovine peripheral blood mononuclear cells (PBMCs) was evaluated. The infectious titres of extracellular virus were measured and the intracellular viral RNA levels were determined by reverse transcription quantitative PCR after cell inoculation. The effects of EV-E on cell viability and proliferative response were investigated with a methyl thiazolyl tetrazolium bromide reduction assay, the percentages of main lymphocyte subsets and oxidative burst activity of blood phagocytes were determined with flow cytometry, and pro-inflammatory cytokine secretion was measured with an ELISA. Results Enterovirus E productively infected bovine PBMCs. The highest infectious dose of EV-E decreased cell viability and T-cell proliferation. All of the tested doses of virus inhibited the proliferation of high responding to lipopolysaccharide B cells and stimulated the secretion of interleukin 1β, interleukin 6 and tumour necrosis factor α pro-inflammatory cytokines. Conclusion Interactions of EV-E with bovine immune cells may indicate potential evasion mechanisms of the virus. There is also a risk that an infection with this virus can predispose the organism to secondary infections, especially bacterial ones.

Aujeszky’s disease, or pseudorabies, is a viral infection caused by Suid herpesvirus 1 (pseudorabies virus), with swine as its natural host. Although eradicated in domestic pigs in many European countries, PRV remains endemic in wild boar populations, posing a risk to other species, including carnivores. In this study, we report eight fatal cases of PRV infection in hunting dogs from Epirus and Thessaly, Greece, all of which followed direct contact with hunted wild boars. Postmortem brain samples tested positive for PRV via PCR targeting the glycoprotein C (gC) gene. Partial sequencing and phylogenetic analysis of the amplified gC fragments revealed genetic divergence among the examined isolates. The Epirus-derived strains formed a distinct cluster, closely related to previously reported Greek strains from the region of Central Macedonia as well as to the French strain FRA 527 and the German isolate GER614BW. In contrast, the two Thessaly sequenced isolates were phylogenetically distant from all other Greek strains, potentially representing an independently evolving lineage, and clustered more closely with the Kaplan strain. These findings underscore the persistent threat of PRV transmission from wild to domestic species and highlight the genetic heterogeneity of PRV strains circulating in Greece. Veterinary practitioners should consider PRV in the differential diagnosis of encephalitic symptoms in hunting dogs. Enhanced molecular surveillance and public awareness are critical to mitigating the risks posed by this emerging threat.

Rheumatoid arthritis is often accompanied by neuropsychiatric manifestations, suggesting interactions between systemic inflammation and the central nervous system. This study evaluated the effects of sertraline, compared with methotrexate, infliximab, and tocilizumab, on circulating inflammatory and neurotrophic mediators and the striatal expression of selected CNS-related genes in collagen-induced arthritis. Male Wistar rats were assigned to seven groups: adjuvant control, untreated arthritis, methotrexate, sertraline, methotrexate plus sertraline, infliximab, or tocilizumab. At week 12, circulating IL-6, IL-15, IL-10, BDNF, myostatin, and irisin were measured, and striatal BDNF, IL-1β, and GFAP expression was assessed by quantitative real-time PCR. IL-6 concentrations differed between groups, with the lowest values after infliximab treatment and the highest in untreated arthritic rats and animals receiving methotrexate plus sertraline or tocilizumab. IL-15 was highest in untreated arthritis, whereas IL-10 was generally higher in treated groups and peaked after tocilizumab. The IL-6/IL-10 ratio was lower in all treated groups than in untreated arthritis, with the most pronounced reduction observed after infliximab. Circulating BDNF concentrations were highest in the infliximab- and tocilizumab-treated groups. Striatal BDNF expression differed significantly between groups, whereas no significant between-group differences were detected in striatal GFAP or IL-1β expression. No significant between-group differences were observed for circulating myostatin or irisin. Sertraline-containing regimens were associated with lower IL-6/IL-10 ratios than untreated arthritis, whereas infliximab and tocilizumab were associated with the most pronounced changes in systemic inflammatory and neurotrophic parameters.