Epizootic hemorrhagic disease (EHD) is an important disease that severely affects wild and domestic ruminants, and causes substantial economic losses in the livestock industry. The etiological agent, epizootic hemorrhagic disease virus (EHDV), is an arbovirus of the genus Orbivirus. The virus is transmitted through infected Culicoides midges biting susceptible hosts. EHDV has a worldwide distribution, and recently, it has expanded to Europe from 2022. To date, seven different EHDV serotypes have been described and no effective DIVA (Differentiating Infected from Vaccinated Animals) vaccines against EHDV are available. The aim of our research is to explore vaccine candidates expressing a single EHDV antigen to develop an efficient marker and safe vaccine candidate. Recombinant Modified Vaccinia Ankara virus (MVA) viral vector vaccines are effective, safe, and DIVA-compatible candidates against EHDV. In this study, we designed and generated novel vaccine candidates against EHDV based on recombinant MVAs that express the non-structural protein NS1 from EHDV serotype 8. The potential vaccine was evaluated in IFNAR (-/-) mice, which are susceptible to EHDV infection and reproduce certain features of EHDV infection in natural hosts. IFNAR (-/-) mice were immunized with two doses of MVA-NS1 and subsequently challenged with EHDV-8 or EHDV-6. Immunized mice did not show viremia or clinical signs after EHDV-8 challenge, and showed reduced RNAemia titers against the virus. Therefore, vaccinated mice were completely protected against a homologous challenge with EHDV-8 after two doses of the vaccine candidate. MVA-NS1 did not confer complete protection against a heterologous challenge with EHDV-6, although it delayed the onset of clinical signs and death. These data show that the MVA-NS1 vaccine candidate is safe and efficient against a homologous challenge and compatible with a DIVA strategy.

Rift Valley fever virus (RVFV; Genus Phlebovirus, Family Phenuiviridae, Order Hareavirales) is a mosquito-borne arbovirus that affects ruminants and humans. The RVFV genome consists of three single-stranded, negative-sense RNA segments: S (Small), M (Medium), and L (Large). The lack of a licensed vaccine with an optimal safety profile remains a major challenge for disease control. Previously, we generated an attenuated RVFV variant, 40Fp8, through random mutagenesis of the virulent South African strain 56/74. This variant showed a hyper-attenuated phenotype and an optimal safety profile in both mice and pregnant sheep, making it a strong candidate for a live attenuated vaccine (LAV). However, LAVs may pose risks in nature due to potential reversion to virulence and/or reassortment events in the case of segmented viruses. In this study, we analyzed the contribution of each 40Fp8 genomic segment to attenuation using a reverse genetics system. Recombinant viruses were generated by replacing one genomic segment of 56/74 with the corresponding segment from 40Fp8. In vitro, viruses carrying the 40Fp8 M segment displayed a small-plaque phenotype and higher replication kinetics like 40Fp8, whereas those with the 40Fp8 S or L segments behaved like the parental 56/74 strain. In vivo, sex-dependent differences were observed, with male mice more susceptible than females, likely due to hormonal, immunological, and genetic host factors. Notably, the virus carrying the 40Fp8 S segment conferred complete protection in both sexes and at all tested doses. Our data indicate that the 40Fp8 S segment is the primary determinant of attenuation, followed by the M and L segments, which provided partial protection, suggesting that coinfection events of circulating virulent RVFVs with 40Fp8 would likely preserve an attenuated phenotype and supporting the notion that the use of the 40Fp8 as an LAV would carry minimal risk in the event of reassortment in nature.

Porcine circovirus type 2 (PCV2) compromises the immune system in pigs, increasing their susceptibility to co-infections of other pathogens. Classical swine fever virus (CSFV) is one of those diseases and is listed as a highly contagious disease by the World Organization for Animal Health (WOAH). Effective vaccination is one of multipleeffective strategies to prevent diseases and requires the stimulation of T helper cell-mediated immune responses to enhance the effectiveness of vaccines. We developed a bivalent subunit vaccine containing PCV2 ORF2 and CSFV E2 antigens, formulated with a porcine-specific CpG adjuvant. Those specific pathogen-free piglets were immunized with only a single dose and challenged with virulent strains of PCV2 or CSFV four weeks post-vaccination. Vaccinated piglets had significantly higher neutralizing and ELISA antibody titers against both viruses. Improved cellular immunity is demonstrated by increasing percentages of CD3⁺CD4⁺CD8⁺ T cells and significantly upregulated mRNA expression of IL-2, IFN-α, and IFN-γ in peripheral blood mononuclear cells in vaccinated pigs after a challenge. No clinical signs or lesions were observed in vaccinated pigs, and viral loads in serum and tissues were markedly reduced or undetectable. In conclusion, a single dose of the PCV2/CSFV bivalent subunit vaccine induces both robust humoral and cellular immune responses, offering strong protection against infection. These findings highlight its potential as a practical and effective strategy for swine disease control.

Non-communicable diseases (NCDs) are a growing concern not only in human populations but also in animals, with significant implications for ecosystem health and human well-being. The interconnection of animal NCDs, environmental factors, and human health, highlights the role of the One Health framework in understanding and addressing these complex interactions. Animals, particularly livestock and wildlife, are increasingly affected by NCDs, such as obesity, diabetes, cancers, and cardiovascular diseases, often mirroring human health trends. Environmental factors, including pollution, climate change, and habitat degradation, are connected with the emergence and progression of these animal diseases. Exposure to environmental toxins, such as pesticides and heavy metals, has been linked to metabolic disorders and cancers in both animals and humans. Additionally, shared risk factors, such as sedentary lifestyles and poor nutrition, further fortifythe connection between animal and human NCDs. The implications of animal NCDs extend beyond animal health, affecting food security, zoonotic disease dynamics, and ecosystem stability, ultimately impacting human health. Thus, the study and management of NCDs in animals are essential not only for ensuring animal welfare and livestock production but also for contributing to the broader One Health framework. A One Health approach is critical to address the root causes of animal NCDs, advocating for integrated strategies that promote sustainable environmental practices, improve animal husbandry, and enhance surveillance systems, ultimately leading to the development of holistic solutions to mitigate the burden of NCDs across species and ecosystems, fostering a healthier planet for all.

Staphylococcus spp. commonly colonize the skin, mucosal surfaces, and gastrointestinal tracts of humans and animals, among which Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), represents a pathogen of concern. Although MRSA is traditionally associated with healthcare settings, its increasing prevalence in the community has raised concerns about its transmission from animals to humans. Studies comparing domestic and feral cats demonstrate that indoor pet cats exhibit higher colonization rates of S. aureus, approximately 19% vs. 8% among feral cats, and MRSA carriage reaches about 10% in domestic cats, compared to only 1.4% in feral populations. In healthy indoor cats, MRSA prevalence averages 6.6%, with risk factors including households where owners work in healthcare, co‑resident dogs, and antibiotic treatment in the past year. Although feline stomatitis has not been directly studied for MRSA prevalence, existing data show that oropharyngeal staphylococci from cats often demonstrate high resistance rates ~99% to at least one antibiotic, ~12% multidrug resistance, and frequent carriage of resistance genes including mecA. Microbiome research in cats with gingivostomatitis, periodontitis, and chronic stomatitis reveals significant oral dysbiosis characterized by enrichment of anaerobic periodontal pathogens (Treponema, Porphyromonas) and reduced commensal taxa, thereby creating conditions favorable to opportunistic staphylococcal colonization. Behavioral patterns, such as self‑licking and human face/hand licking, further increase the opportunity for the bidirectional transmission of MRSA between cats and owners. Moreover, while MRSA prevalence in cats with stomatitis is yet to be directly quantified, the convergence of high antibiotic exposure, documented indoor MRSA carriage, and oral microbial disruption strongly suggests that cats suffering from chronic oral inflammatory diseases and undergoing repeated or prolonged antibiotic therapy may act as overlooked reservoirs of MRSA. Understanding the role of feline S. aureus in MRSA epidemiology is crucial for assessing potential risks to human cohabitants and improving public health strategies.

Swine influenza (SI) is an acute infectious disease of pigs caused by Swine influenza virus (SIV). To develop virus-like particles (VLPs) of SIV H1 and H3 subtype and determine their morphological structure and biological characteristics, this study synthesized a gene encoding lumazine synthase (LS) derived from a hyperthermophilic bacterium and introduced an immunoglobulin-binding domain sequence from streptococcal protein G (pG) to construct the pG-LS recombinant plasmid. The plasmid was then expressed in both prokaryotic and eukaryotic systems.After the conjugation of the 2 proteins, the morphology and the formation of VLP could be observed by transmission electron microscopy. Average particle diameters of the conjugated proteins were measured with dynamic light scattering method while the biological properties of the complex protein were analyzed by SDSPAGE, Western blot and hemagglutination assay. Both expression methods successfully yielded pG-LS nanoparticles, with eukaryotic expression demonstrating higher purity. The complexes formed between the two HA-Fc proteins and eukaryotic-expressed pG-LS nanoparticles exhibited a spherical morphology with an average diameter of approximately 80 nm.Further immunological analysis and particle diameter measurement both showed the successful construction of VLPs. Hemagglutination assays indicated HA-Fc alone could not agglutinate erythrocytes, while the virus-like particles prepared from 2 conjugated proteins showed exponential affinity, which could efficiently agglutinate red blood cells of different species. This study successfully constructed pG-LS nanoparticles and 2 pG-LS-HA VLPs, where HA proteins successfully displayed at the VLP surface. Both developed pG-LS-HA had stable property and biological function, with a potential of large-scale production. In the meantime, this study found that both H1 and H3 HA could be effectively displayed to form VLP, which shed light on the future use of multi-subtype influenza virus HA protein co-presentation strategy to develop novel, multivalent VLP vaccine candidates.

African swine fever (ASF) is a highly lethal transboundary disease that severely impacts the swine industry. Because of the complexity of the ASF virus (ASFV), vaccine development remains challenging, highlighting the need to robustly evaluate immune responses induced by vaccine candidates and their efficacy against different viral strains. This study evaluates the utility of the hemadsorption inhibition assay (iHAD) as an in vitro method to evaluate ASFV specific immune responses, and compares its results with in vivo protection outcomes.

Serum samples were selected from wild boars involved in two separate in vivo immunization trials against the genotype II isolate Arm07. In the first trial, animals were immunized with the naturally attenuated and non-hemadsorbing strain Lv17/WB/Rie1 (Rie1), followed by a challenge with the heterologous strain Ken06.bus. In the second trial, animals were immunized with the gene-deleted vaccine candidate Lv17/WB/Rie1ΔAB (dAB). In both trials, vaccinees exhibited high protection levels against Arm07, and selected sera demonstrated high ASFV-specific antibody titers. The inhibitory activity of these sera was tested by iHAD against homologous isolates (Arm07, Rie1, dAB) and heterologous isolates (Ken06.bus, Esp70, Ken07).

90% of the animals were protected in vivo against Arm07, while 30% of the sera exhibited in vitro iHAD. For the heterologous strain Ken06.bus, 30% of the animals showed in vivo protection, whereas 80% of the sera demonstrated in vitro inhibitory activity. Sera from animals previously immunized with genotype II vaccine strains inhibited hemadsorption by the virulent strains Arm07, Ken06.bus, and the moderately virulent strain Esp70.

The iHAD correlates with the inhibitory capacity of sera from vaccinated animals but cannot be considered a definitive marker of vaccine efficacy for Rie1 and dAB strains. The in vitro results suggest the existence of complete or partial cross-protection among phylogenetically distinct ASFV strains, including Arm07, Ken06.bus, and Esp70.

Brucellosis-free status in Zhijiang County, Hubei, is jeopardised by the continual inflow of live cattle from high-prevalence provinces. This study integrated participatory value-chain mapping with stochastic risk modelling to quantify where and how Brucella could be introduced.

Two focus-group discussions, 48 key-informant interviews, and 31 site visits delineated the cattle production-to-consumption network, characterising trade volumes, stakeholder practices, and biosecurity gaps. These data parameterised a scenario-tree model run in @Risk (50,000 Monte Carlo iterations), and a Latin-hypercube global sensitivity analysis ranked the influence of input variables.

Brokers dominated live-animal movements, sourcing ~80 % of imports and frequently bypassing origin-quarantine. The median probability that an individual imported feeder was Brucella-positive was 1.43 % (95 % CI: 0.68–2.18), yielding an annual probability >99.999 % that at least one infected animal enters Zhijiang in 95 % of simulations. Over 90 % of total risk stemmed from cattle introduced via (i) unregulated brokers or (ii) trading platforms that omitted origin-quarantine or relied on low-sensitivity pen-side tests. Origin-herd prevalence and the proportion of cattle handled by brokers were the most influential parameters.

Even under conservative assumptions, current movement controls cannot prevent brucellosis incursion. Policy priorities include enforcing origin-quarantine, accrediting brokers, instituting centralized post-entry isolation, and deploying high-specificity rapid diagnostics. By coupling qualitative value-chain insights with quantitative risk assessment, this framework offers a transferable template for evidence-based disease-freedom programmes and highlights critical intervention points for safeguarding public health and livestock economies.

Canine enteric coronavirus (CeCoV) is one of the most prevalent viruses in dogs, resulting in gastrointestinal signs and, in severe cases, death. Despite being a major disease, study on its spread remains limited, particularly in some geographical areas, such as the Campania region (southern Italy). This study investigated the CECoV serological and molecular occurrence, as well as the risk factors related to increased exposures. A total of 258 blood and 154 fecal samples were obtained from dogs in 71 districts, accompanied by information such as sex, breed, size, location, age, origin, lifestyle, and attitude. The serological and molecular prevalences were obtained using a commercial ELISA (also useful for evaluating the antibody titer) and a previously described real-time PCR protocol, respectively. More than half of the dogs investigated (53.9%) had been exposed to the pathogen, while only 5.8% tested positive in real-time PCR. Among the animals that tested seropositive, 57 had a titer of 1:50, 36 of 1:100, and 46 of 1:200. Univariate and multivariate studies found that hunting dogs and dogs with outdoor lifestyles had greater seroprevalence. Sex, age, breed, origin, and size were not associated with higher seroprevalences. Furthermore, dogs in some areas exhibited seroprevalences of up to 85%. The same area (Avellino and Salerno) included most of the PCR-positive animals. Among these nine animals, only two were seronegative when tested by ELISA, another five had a titer of 1:50, and two of 1:200. Furthermore, eight out of nine animals were hunting dogs or strays, only one was a pet dog, and seven/nine lived outdoors. The study's findings highlighted the extensive prevalence of CECoV in the examined area, as well as the presence of ongoing outbreaks in many districts. More efforts are needed to offer useful epidemiological surveillance for this important canine disease, as well as to detect mutations and to anticipate possible changes in its epidemiological cycle.

Notoedric mange, caused by Notoedres cati, affects cats and some rodent species, less commonly dogs and foxes. Young kittens are usually affected. It also causes transient lesions in humans. We report a case of generalized notoedric mange, in an adult tomcat (six years old) consequently to immunosuppression induced by infection with feline immunodeficiency virus (FIV). This is a retrovirus that causes suppression of cellular immunity. The disease manifested as extensive, very itchy crusted dermatitis, deep pyodermatitis on the limbs, and mucopurulent conjunctivitis. The disease is highly contagious. N. cati belongs to the family Sarcoptidae, with a life cycle and morphology very similar to Sarcoptes canis. Diagnosis can be clinically oriented. The distribution of lesions and intensity of pruritus are highly suggestive. Confirmation of mange was made by microscopic examination of the skin scrapings, the parasites being more abundant than in sarcoptic mange. Confirmation of FIV was made with Rapid Antigen FIV Ab/ FeLV Ag Test Kit, Bionote Inc. (Gentaur Group Europe). FIV infection progresses asymptomatically for a long time, being underdiagnosed. Once immunosuppression develops, it predisposes to numerous infectious or parasitic diseases. It is often associated with lymphoplasmacytic stomatitis. Reported dermatological signs include chronic and recurrent bacterial skin infections and otitis, an increased frequency of infection with Cryptococcus neoformans, Candida albicans, or Microsporum canis, and demodicosis.