Mycoplasma synoviae (MS) is a key avian pathogen that causes respiratory, joint, and reproductive issues in poultry, resulting in significant economic losses. This study aimed to determine the seroprevalence and pathological changes linked to MS infection in commercial broiler flocks in District Karak, Khyber Pakhtunkhwa, Pakistan. A total of 200 blood samples were collected from 13 broiler farms and tested using an indirect ELISA kit to detect MS-specific antibodies. Additionally, tissue samples from the trachea and lungs were taken from naturally infected birds and examined histopathologically. The serological results showed that 41 out of 200 samples (20.5%) tested positive for MS antibodies, indicating moderate pathogen circulation in the area. Risk factor analysis revealed higher seroprevalence in birds over three weeks old (23%) compared to those under three weeks (12%), and in smaller flocks (<2000 birds; 25%) versus larger flocks (>2000 birds; 19%). However, statistical analysis (Chi-square test) found no significant associations (p > 0.05) with age, flock size, feed, or mortality. Clinical signs like respiratory distress, anorexia, ascites, and white diarrhea were weakly linked to seropositivity. Gross lesions included tracheal hemorrhages, splenomegaly, synovitis, and lung consolidation. Histopathology showed epithelial desquamation, leukocytic infiltration, hemorrhage in the trachea, and alveolar septal thickening and congestion in the lungs, consistent with septicemia progression of MS infection. This study confirms that Mycoplasma synoviae is endemic in broiler flocks of District Karak, with characteristic respiratory lesions. The findings highlight the importance of regular surveillance, biosecurity, and improved diagnostic methods to reduce the impact of MS on poultry production in Pakistan.

Tick-borne diseases (TBDs) are significant health concerns affecting equines worldwide, causing a range of clinical syndromes and negatively impacting animal welfare and productivity. These diseases are caused by a broad spectrum of tick-transmitted pathogens, including bacteria (e.g., Anaplasma spp., Rickettsia spp.), protozoa (e.g., Babesia caballi, Theileria equi), and viruses such as tick-borne encephalitis virus. Sicily, Italy, with its Mediterranean climate and widespread ticks, represents a potentially high-risk area for equine TBDs. This study aimed to investigate the circulation of major tick-borne pathogens in equines (horses and donkeys) to improve understanding of their epidemiological status and to support targeted control strategies.

Between 2024 and 2025, blood, serum, and ticks were collected from equines. DNA was extracted from whole blood and ticks and subjected to PCR assays targeting genes of key pathogens, including Rickettsia spp. (ompB, ompA, gltA), Anaplasma spp. (16S rRNA), Borrelia burgdorferi (OspA), Babesia caballi (48 kDa rhoptry protein), and Theileria equi (EMA-1). Serum samples were analyzed by indirect immunofluorescence assay (IFA) for antibodies against Anaplasma spp. and B. caballi, and by ELISA for flavivirus antibodies.

Of the 163 blood samples tested, 16% were positive for T. equi, while no other pathogens were detected. A total of 29 ticks were collected: 20 Ixodes ricinus, 6 Haemaphysalis punctata, and 3 nymphs. One Haemaphysalis tick tested positive for Anaplasma spp. Among 196 equine serum samples, 12.2% were seropositive for Anaplasma phagocytophilum, 3.8% for B. caballi, and 6% for flaviviruses.

The molecular detection of tick-borne pathogens in equines from Sicily confirms the circulation of multiple TBD agents in the region. These findings highlight the need for improved surveillance and implementation of tick control measures to manage equine TBDs. Further studies are warranted to evaluate clinical relevance and local vector ecology.

This research was funded by the Italian Ministry of Health: IZSSI 03/23 RC.

Vector-borne pathogens such as Rickettsia spp. and Anaplasma spp. are increasingly recognized as emerging threats to human and animal health. Domestic cats (Felis catus) may act as sentinels, reservoirs, or incidental hosts, especially in Mediterranean areas where arthropod vectors are widespread. This study aimed to investigate exposure and infection rates of Rickettsia spp. and Anaplasma spp. in cats from Sicily (Italy), including molecular screenings of their ectoparasites.

Serum samples were tested for antibodies against Rickettsia spp. and Anaplasma spp. using indirect immunofluorescence assay (IFA). DNA extracted from EDTA blood and ectoparasites (ticks and fleas) was subjected to PCR for Rickettsia spp. and Anaplasma spp. Positive PCR products were sequenced to confirm species identity.

Between 2024 and 2025, samples were collected from 80 cats. Serological testing revealed high exposure rates, with 56.3% of cats positive for Rickettsia spp. antibodies and 12.7% for Anaplasma spp. In contrast, molecular analyses detected Rickettsia felis DNA in only 3.8% of feline blood samples, while no Anaplasma spp. DNA was identified. Among ectoparasites, R. felis DNA was detected in one of two fleas (Ctenocephalides felis), and Anaplasma phagocytophilum DNA in one (Ixodes ricinus) of eight ticks.

The marked discrepancy between seroprevalence and molecular detection suggests that while cats are frequently exposed to these pathogens, active infections detectable by PCR are uncommon, likely due to a transient bacteremia that limits the window of molecular detectability. The identification of pathogen DNA in ectoparasites supports their circulation in the environment, although this alone does not confirm a reservoir role for cats. These findings highlight the importance of the integrated surveillance of companion animals and their ectoparasites within a One Health framework to better understand and mitigate zoonotic risks.

This research was funded by the Italian Ministry of Health (IZSSI 03/23 RC).

_{The increasing emergence of antimicrobial-resistant pathogens, including Methicillin-Resistant Staphylococcus aureus (MRSA), poses a significant threat to food safety and public health. The unhygienic handling practices combined with environmental contamination and antibiotic misuse in Lagos and Nigeria's coastal communities make crustacean seafood a potential reservoir for MRSA. A total of 71 crustacean seafood samples were randomly collected from three major seafood markets across Lagos State. Baird Parker Agar served as a medium for S.aureus isolation, followed by Gram staining and biochemical tests to confirm the results. The Kirby–Bauer disc diffusion method on Mueller–Hinton agar followed CLSI guidelines to test presumptive isolates for antibiotic susceptibility. The phenotypic marker tests using Cefoxitin(30 µg) and Oxacillin(5 µg) identified MRSA strains. The presumptive S. aureus isolates from 71 samples reached 60 (84.5%), and the phenotypic resistance tests showed 22 (36.67%) and 60 (100%) isolates were resistant to cefoxitin and oxacillin, respectively. Further, 6 out of 21 processed isolates showed resistance to cefoxitin and all 21 processed isolates showed resistance to oxacillin. The 39 fresh isolates showed 16 cefoxitin-resistant strains but all 39 samples displayed complete resistance to oxacillin.}

_{The antibiotic susceptibility profile revealed that each of Augmentin (100%), Meropenem (100%), Ceftriaxone (100%) and Linezolid (100%) showed high resistance, while ciprofloxacin exhibited relatively lower resistance. The research demonstrates that MRSA exists in crustacean seafood products sold in Lagos, which creates potential health risks for both consumers and handlers. The high level of MDR indicates that antibiotics have been used either excessively or improperly in aquatic environments and post-harvest handling processes. The recommended measures to control resistant strain spread include regular surveillance and proper seafood handling practices, together with controlled antimicrobial use in aquaculture operations. The research provides essential local findings about MRSA in seafood, which supports the requirement for One Health-based AMR monitoring in Nigeria.}

Tick-borne diseases pose a serious challenge to cattle farming in Mediterranean endemic areas. In Sicily, pathogens such as Anaplasma marginale, Babesia spp., and Theileria annulata are widespread and can impact herd health and productivity. This study was conducted on a farm in Sicily, Italy, where non-native cattle imported from Austria, the Netherlands, and France showed severe clinical signs likely related to tick-borne infections.

Among the 35 cattle on the farm, 26 were tested for antibodies to A. marginale, Babesia bigemina, and Crimean-Congo Hemorrhagic Fever virus (CCHFV) by ELISA, and for Babesia bovis and T. annulata by indirect immunofluorescence assay (IFA). PCR was performed on whole blood and spleen (from one dead animal) for A. marginale, B. bigemina, B. bovis, and T. annulata. Ticks were identified morphologically and molecularly and screened for Rickettsia spp. Amplicons were sequenced and compared to GenBank.

Serology showed 21/26 positive for A. marginale, 20/26 for B. bovis, 3/26 for B. bigemina, and 14/26 for T. annulata; none were positive for CCHFV. PCR confirmed A. marginale (9/26), B. bigemina (1/26), and T. annulata (1/26); B. bovis was not detected. The spleen tested positive for A. marginale and T. annulata. Ticks were identified as Hyalomma lusitanicum, PCR-positive for Rickettsia aeschlimannii.

The introduction of non-native cattle into tick-endemic areas increases the risk of disease. High seroprevalence of A. marginale and T. annulata indicates widespread exposure. Differences between serology and PCR reflect past exposure versus active infection. H. lusitanicum is a known CCHFV vector in Mediterranean regions. However, no CCHFV seropositivity was detected, suggesting no active viral circulation in the herd. Still, the established presence of this tick species in Sicily, along with climate change and animal movement, could favor local emergence. Enhanced surveillance is recommended to support early outbreak detection and protect animal and public health.

SIGLEC15, a critical immune receptor in chickens, plays a crucial role in immune responses to infections, including Newcastle Disease (ND). This study aims to explore the potential of polyphenols as inhibitors of SIGLEC15-mediated Newcastle Disease Virus (NDV) binding to enhance immunity in chickens. Polyphenols are bioactive plant-derived compounds known for their ability to modulate immune receptors and influence signaling pathways. Molecular docking was conducted on 12,000 polyphenols to identify those with high binding affinity for SIGLEC15. Among the screened compounds, Quercetin and Resveratrol demonstrated superior docking scores (-9.45 kcal/mol and -8.79 kcal/mol) and lower RMSD values (0.58 and 0.71, respectively) compared to synthetic SIGLEC15 inhibitors. These polyphenols effectively disrupted the binding sites of NDV on SIGLEC15, inhibiting critical receptor–ligand interactions necessary for viral entry. The NF-κB signaling pathway, activated by SIGLEC15–ligand interactions, was identified as a key pathway modulated by polyphenols. Molecular dynamics simulations (MDS) confirmed the stability of polyphenol–SIGLEC15 complexes, indicating sustained interactions that block NDV invasion. Pharmacokinetic analysis showed favorable profiles, including non-toxicity, high bioavailability, and potential for immune enhancement. By targeting SIGLEC15 and interrupting NDV binding through the NF-κB pathway, these findings provide a novel strategy to improve immunity and protect chickens from Newcastle Disease Virus.

Introduction:
Porcine delta coronavirus (PDCoV) and porcine epidemic diarrhoea virus (PEDV) continue to pose a threat to global swine health, animal welfare, and food security. These enteric coronaviruses are associated with severe diarrhoea, dehydration, and high mortality in neonatal piglets. Despite significant economic losses, no antiviral treatments are currently approved, and vaccine control remains suboptimal due to viral evolution and incomplete protection. Repurposing clinically approved drugs that target conserved viral proteases represents a rapid and cost-effective therapeutic strategy aligned with the principles of One Health.

Methods:
We retrieved high-resolution crystal structures of PDCoV and PEDV 3CL proteases (PDB IDs: 4XFQ and 8E7C). Following the validation of docking protocols using AutoDock Vina and PyRx software, compound libraries from DrugBank and ZINC15 were virtually screened. Top-ranking hits were prioritised based on docking scores and interaction profiles. These candidates were subjected to molecular dynamics simulations to evaluate complex stability over time. In silico pharmacokinetic and toxicity predictions were performed using SwissADME and pkCSM tools to assess oral bioavailability and safety profiles.

Results:
Saquinavir, an FDA-approved HIV protease inhibitor, demonstrated consistently favourable docking interactions with conserved catalytic residues in the viral protease active sites. Molecular dynamics simulations confirmed the stability of binding conformations over 100 ns, supporting the robustness of the predicted interactions. Pharmacokinetic profiling indicated acceptable oral absorption and low predicted toxicity.

Conclusions:
This integrated virtual screening and simulation pipeline identifies saquinavir as a promising repurposed candidate against porcine enteric coronaviruses. These findings justify urgent in vitro validation and illustrate how drug repurposing can accelerate antiviral discovery in veterinary medicine within a One Health framework.

Highly pathogenic avian influenza (HPAI) is a devastating viral disease that leads to severe pulmonary damage and high mortality rates in domestic poultry worldwide. It is caused by avian influenza type A viruses from the Orthomyxoviridae family. The HPAI virus contains a genome of eight negative-sense RNA segments. Among these, the PB2 (Polymerase basic protein) segment plays a crucial role in viral replication as a subunit of the viral RNA-dependent RNA polymerase (RdRp) complex. Targeting PB2 expression represents a potential strategy to control HPAI. MicroRNAs (miRNAs), which are short noncoding RNAs, play a significant role in post-transcriptional regulation of gene expression, influencing various biological processes including cell growth, tissue differentiation, apoptosis, and viral infection. Our previous research identified 200 differentially expressed pulmonary miRNAs in chickens during HPAI infection. In the current study, we screened these dysregulated miRNAs against the PB2 segment of the HPAI H5N1 virus. We evaluated parameters such as the thermodynamic stability of miRNA-mRNA complexes, complementarity of miRNA seed sequences with mRNA, conservation of the PB2 sequence, and accessibility of target sites to identify miRNAs that could potentially target the PB2 transcript of H5N1. Our findings indicate that chicken miRNAs gga-miR-17-3p, gga-miR-29a-5p, gga-miR-1718, gga-miR-16c-5p, and gga-miR-1744-5p are predicted to target the PB2 segment. This suggests that chickens possess inherent genetic potential to combat H5N1 infection, which can be leveraged to develop effective control strategies for HPAI in poultry and miRNA therapeutic potential.

Introduction:
Newcastle Disease (ND) is a highly contagious viral infection that continues to cause significant mortality, production losses, and trade restrictions in poultry worldwide. Despite widespread vaccination, outbreaks persist due to the emergence of more virulent Newcastle Disease Virus (NDV) strains and reduced vaccine efficacy under field conditions. Complementary antiviral strategies are increasingly recognized as a necessary addition to vaccination programs.

Methods:
A comprehensive literature review was conducted using databases including PubMed, Web of Science, and Scopus. Studies published in English up to 2025 were screened for evidence on synthetic antivirals, plant-derived compounds, immunomodulators, and supportive agents against NDV. Both in vitro and in vivo studies were evaluated, with a focus on antiviral efficacy, safety, cost, and practical field applicability.

Results:
Synthetic antivirals, such as ribavirin and favipiravir, demonstrated moderate efficacy in controlled laboratory studies but are limited by high cost, toxicity concerns, and regulatory restrictions in poultry. Plant-based compounds, notably Curcuma longa (turmeric), Azadirachta indica (neem), and Allium sativum (garlic), showed promising antiviral and immunostimulatory effects with better accessibility and safety profiles. Immunomodulators, including interferons, and supportive agents such as vitamins and probiotics, enhanced host resistance. However, most evidence comes from in vitro studies, with a shortage of well-designed in vivo and field trials.

Conclusions:
The literature supports integrating validated plant-based antivirals and immunomodulators with vaccination to create a more robust and sustainable ND control strategy. Priority research should focus on large-scale in vivo and field evaluations to confirm efficacy, safety, and cost-effectiveness under commercial conditions. Such integration could significantly reduce ND-related losses and improve poultry health management.

Swine acute diarrhea syndrome coronavirus (SADS-CoV), an emerging alphacoronavirus, leads to fatal diarrhea in piglets and exhibits broad cross-species transmission potential. Bone marrow stromal cell antigen 2 (BST2) is an IFN-induced host protein known to restrict a wide range of enveloped viruses. Previous studies have shown that BST2 inhibits Porcine epidemic diarrhea virus (PEDV) replication through the selective autophagy pathway, suggesting a novel mechanism for its antiviral activity. Given that SADS-CoV and PEDV are phylogenetically related alphacoronaviruses, we hypothesized that BST2 may employ a similar mechanism to antagonize SADS-CoV infection.

To test this hypothesis, we employed TurboID-based proximity labeling combined with co-immunoprecipitation and Western blot analysis. Our results demonstrated that BST2 binds to the SADS-CoV nucleocapsid (N) protein and promotes its degradation, resulting in a significant reduction in viral replication. Further virological assays showed that overexpression of BST2 significantly reduced viral titers, whereas BST2 knockout enhanced viral replication.

These findings establish BST2 as a key host restriction factor against SADS-CoV and reveal a novel antiviral mechanism targeting the viral nucleocapsid. This work advances our understanding of innate immune defenses against coronaviruses and highlights BST2’s potential as a target for developing interventions aimed at enhancing antiviral responses in swine and mitigating zoonotic risks.