The emergence of mpox cases in Ireland in May 2022 triggered an urgent public health response, including an attempted analytical epidemiological investigation aimed at informing vaccination strategies and risk communication, particularly among gay, bisexual and other men who have sex with men. At the time of implementation, case numbers had declined to an extent that limited recruitment, resulting in early suspension of data collection. We present a structured reflection of the investigation, drawing on a lessons learnt review and the experiences of those involved, to offer actionable recommendations to strengthen evidence generation in future outbreak preparedness and response efforts. Key lessons include the critical importance of stakeholder engagement, clear governance and legal frameworks, and the need for enhanced procedural preparedness and pre-established surge capacity arrangements to enable rapid analytical investigations during health emergencies. These insights align closely with Ireland's National Health Protection Strategy and the extended mandate of the European Centre for Disease Prevention and Control. Beyond existing international frameworks for outbreak preparedness, this reflection highlights the practical implementation barriers that must be addressed to enable timely and coordinated epidemiological investigations across Europe.

The monkeypox virus (MPXV) has become a global health threat, with rising cases reported globally. Since its re-emergence in 2017, Nigeria has experienced a sustained outbreak, underscoring the critical need for robust surveillance mechanisms to identify transmission dynamics. Despite this urgency, data regarding MPXV seroprevalence among patients seeking medical care in Nigeria remains scarce. MPXV may be underreported because its clinical symptoms overlap with those of other common causes of fever.

In July 2024, we reported information on the initial human mpox cases in Uganda. Genomic characterization of clinically suspected cases (July-December 2024) revealed cocirculation of several monkeypox virus clade 1b clusters and varicella zoster virus. Our findings highlight the need to refine differential diagnoses by molecular techniques and strengthen surveillance.

During the 2025 mpox outbreak in Sierra Leone, we assessed environmental contamination in 2 hospitals. Of 89 surfaces sampled, 6 (6.7%) surface samples, primarily from doors, were PCR-positive for monkeypox virus. Our findings highlight monkeypox virus surface contamination in healthcare settings and help prioritize infection prevention and control targets in resource-limited settings.

In this meta-analysis, we performed a literature search for articles relating to asymptomatic monkeypox virus (MPXV) detections published worldwide during May 12, 2022-September 13, 2024. The primary outcome was to determine the pooled prevalence of asymptomatic MPXV detections. We conducted a random-effects analysis that yielded a final pooled prevalence of 1.0% (95% prediction interval 0.1%-11.2%). Results did not substantially differ by geographic region or study design. Although those findings are reassuring, we did not evaluate infectivity or transmission potential among asymptomatic persons. MPXV continues to circulate globally; the viral dynamics that enable its persistence remain poorly understood and merit further study.

The global resurgence of mpox, caused by monkeypox virus (MPXV), has highlighted the need for effective antiviral strategies, rapid diagnostics, vaccination, and integrated preparedness approaches. This review critically examines recent advances in MPXV biology, pathogenesis, transmission, clinical manifestations, prevention, diagnosis, treatment, and emerging nanomedicine-based strategies within a One Health framework. Particular emphasis is placed on nanoparticle-enabled vaccines, diagnostics, therapeutics, and wastewater surveillance, highlighting their potential to improve antigen delivery, diagnostic sensitivity, targeted drug delivery, and population-level monitoring. The review also compares conventional and nanoscale approaches and evaluates the current evidence supporting their efficacy and translational potential. Importantly, most nano-enabled MPXV interventions remain at the preclinical stage, and their reported performance cannot be directly compared because of differences in nanoparticle composition, target antigens, experimental models, assay conditions, and clinical validation strategies. Major challenges include long-term safety, pharmacokinetics, reproducibility, scalability, storage stability, regulatory approval, and the limited availability of well-designed clinical studies. Within the One Health framework, effective MPXV control also requires coordinated surveillance of human, animal, and environmental reservoirs, including standardized wastewater monitoring and cross-sectoral data sharing. Future research should therefore prioritize rigorous clinical validation, standardized evaluation protocols, diverse MPXV clade testing, long-term safety assessment, scalable manufacturing, and integrated human-animal-environmental surveillance.

The 2022 mpox outbreak highlights the persistent threat posed by the mpox virus (MPXV) and related orthopoxviruses (OPXVs). Yet, approved antibody-based therapeutics for OPXVs are lacking. Here, we present two broadly neutralizing monoclonal antibodies (mAbs), G4 and B9, that target the MPXV M1R protein, homologous to L1R in other OPXVs. Both mAbs demonstrate cross-reactive binding and robust neutralization against multiple pathogenic OPXVs. Notably, G4 exhibits exceptional potency, neutralizing mature virions of VACV with an IC50 of 37.93 pM, whereas B9 has an IC50 of 41.13 nM. Cryo-electron microscopy analysis reveals that G4 and B9 target distinct epitopes on M1R. G4 recognizes a conventional conformational epitope but overcomes neutralization escape associated with the Asp35 residue, whereas B9 binds to an epitope on the lateral β-sheets of M1R. Our findings provide two promising candidates with complementary neutralization profiles for the development of broad-spectrum prophylactic and therapeutic interventions against OPXV infections.

Monkeypox virus (MPXV) has caused two Public Health Emergencies of International Concern since 2022, with co-circulation of distinct clades (Ia, Ib, IIa, IIb) that differ in virulence and transmission patterns. Rapid discrimination of these clades is critical for outbreak control and clinical management. Here, we developed and validated a two-tube multiplex real-time quantitative PCR (qPCR) assay targeting the H3L gene, designed to first distinguish clade I from clade II, followed by subtyping into clades Ia, Ib, IIa, and IIb in separate reactions. The assay demonstrated high sensitivity, with detection limits of 100 copies/reaction for clade Ia, 10 copies/reaction for clade Ib, 10 copies/reaction for clade IIa, and 1 copy/reaction for clade IIb using standard plasmids. Excellent specificity was confirmed by the absence of cross-reactivity with nucleic acids from other human pathogens, including vaccinia virus Tiantan strain (VTT), human adenovirus (HAdV), getah virus (GETV), chikungunya virus (CHIKV), Japanese encephalitis virus (JEV), dengue virus (DENV), influenza A virus (IAV), influenza B virus (IBV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The assay exhibited high repeatability, with intra-assay coefficients of variation (CVs) ranging from 0.09 to 1.88% and inter-assay CVs from 0.23 to 2.68%. Clinical evaluation using 48 skin samples from infected crab-eating monkeys or SCID mice showed 100% concordance with singleplex qPCR for clade IIb detection. For clades Ia, Ib, and IIa, clinical validation was not feasible due to sample unavailability, and their performance was analytically confirmed using recombinant plasmids. Importantly, the clade assignment of the 48 animal specimens was not confirmed by sequencing, which represents a limitation of the current study. Furthermore, these specimens were derived from animal models rather than human patients; thus, the results represent preclinical validation, and formal clinical validation in human specimens from endemic regions is warranted. This two-tube multiplex qPCR assay provides a rapid, sensitive, and reliable tool for the analytical discrimination of all four major MPXV clades, with preliminary clinical validation for clade IIb. Its ability to distinguish emerging clades makes it a valuable asset for clinical diagnosis, epidemiological surveillance, and future outbreak preparedness, although further clinical validation in endemic regions is required.

Mpox virus (MPXV) is an enveloped double-stranded DNA virus belonging to the genus Orthopoxvirus within the family Poxviridae. It causes a zoonotic disease transmitted to humans through direct contact with infected animals or humans, as well as through exposure to contaminated materials. The aim of this study was to conduct a retrospective, comparative diagnostic evaluation of three commercial real-time PCR (qPCR) kits for the diagnosis of MPXV on an enriched, artificially balanced sample set that included 50% MPXV-positive samples. World Health Organization (WHO) reference molecular protocol was used as the reference comparator. A total of 300 clinical specimens, 150 of them MPXV-positive (265 skin lesion/vesicle swabs and 35 serum samples), were used in this analysis. The Bioperfectus kit correctly identified 294/300 samples, showing a sensitivity of 0.96 (95% CI: 0.93-0.99), a specificity of 1.00 (0.98-1.00), and a Cohen's Kappa coefficient of 0.96 (95% CI: 0.92-0.99), with an overall agreement of 0.98 (0.96-0.99). The VIASURE kit correctly detected 293/300 specimens, yielding a sensitivity of 0.95 (0.92-0.99), specificity of 1.00 (0.98-1.00), Cohen's Kappa coefficient of 0.95 (95% CI: 0.92-0.99 and an overall agreement of 0.98 (0.95-0.99). The Singuway kit showed the highest concordance with the reference method, correctly classifying 295/300 samples and presenting a sensitivity of 0.97 (0.94-1.00), specificity of 1.00 (0.98-1.00), Cohen's Kappa coefficient of 0.97 (95% CI: 0.94-1.00), with an overall agreement of 0.98 (0.97-1.00). This study demonstrates a good sensitivity and specificity, and strong overall agreement among the three commercial kits compared to the WHO protocol. Nevertheless, further analysis using unbiased clinical sample sets that reflect the true prevalence of MPXV-positive samples is needed for a real-world clinical performance evaluation.

RNA vaccines-comprising linear mRNA, self-amplifying RNA, and circular RNA (circRNA)-constitute a core next-generation platform for the prevention and control of viral diseases; among these, circRNA vaccines possess notable structural stability, yet their technical bottlenecks and application prospects in veterinary medicine have not been systematically reviewed. This review synthesizes current research on circRNA vaccine design, circularization strategies, translation mechanisms, delivery systems, and immunological outcomes, and compares their antiviral performance with that of linear mRNA vaccines. Owing to their covalently closed circular conformation, circRNA vaccines exhibit enhanced resistance to nucleases and superior thermal stability, enabling sustained transfection activity at ambient temperatures without reliance on strict cold chains; through cap-independent translation driven by internal ribosome entry sites or N[6]-methyladenosine modifications, and in conjunction with optimized circularization protocols and lipid nanoparticle carriers, circRNA vaccines elicit substantially higher antiviral IgG titers and durable antigen-specific T-cell memory relative to linear mRNA vaccines. These vaccines have been deployed against COVID-19, monkeypox, influenza, and livestock viral diseases, demonstrating strong adaptability to viral variants and compatibility with mucosal or needle-free administration routes. CircRNA vaccines are well suited for both emergency outbreak response and routine immunization programs; nevertheless, challenges persist, including low circularization efficiency for long sequences, elevated manufacturing costs, and inadequate quality control standards. Addressing these issues through improved production workflows and delivery technologies adapted to resource-limited settings will be critical to establishing circRNA vaccines as a pillar of livestock disease management and as a strategic reserve for emerging zoonotic threats.

The occurrence of viral zoonotic spillover (the transmission of viruses from animals to humans) has attracted worldwide attention due to mounting concerns regarding viral threats such as avian influenza, Hendra, monkeypox, Nipah and bat coronaviruses. It is evident that these events deviate significantly from natural occurrences, being the conflation of ecological, environmental and social factors that are profoundly altering the boundaries between the animal and human domains. Consequently, in order to comprehend both the contemporary and prospective drivers of zoonotic viral spillover, a coordinated, comprehensive global One Health response is essential. The purpose of the present review is to analyze prevailing pathways of transmission between animal reservoirs and human populations. This examination involves the analysis of historical cases, including those of SARS and Ebola, as well as recent occurrences, such as the global pandemic of SARS-CoV-2 (COVID-19). A secondary objective of this review is to comprehend the fundamental mechanisms of cross-species transmission. The development of effective strategies to mitigate the emergence of zoonotic viruses and prevent future pandemics is contingent on a robust understanding of these mechanisms. This analysis is key to identifying pandemic pathogens and preventing the spread of zoonotic viruses. In conclusion, the review provides a thorough evaluation of current global strategies to prevent the spread of zoonotic viruses, highlighting gaps in our understanding and areas for further research.

The poxviruses are large, epitheliotropic, double-stranded DNA viruses that are pathogens of invertebrates and the mammalian, avian and reptilian vertebrate taxa including the familiar poxviruses of humans and domestic animals such as the variola virus or smallpox virus and the related vaccinia virus used for the live smallpox vaccine, mpox virus (formerly monkeypox virus), cowpox virus of rodents, and orf virus of sheep and goats. Poxviruses have also previously been described from teleost fishes including the common carp Cyprinus carpio, ayu Plecoglossus altivelis, Atlantic salmon Salmo salar, Atlantic cod Gadus morhua, red or Japanese seabream Pagrus major, and black bullhead Ameiurus melas. This report documents a novel poxvirus infection in a captive population of the endangered Knysna seahorse Hippocampus capensis. Histopathological examination revealed an epidermal keratinocytic dermatopathy with hypertrophy and subsequent progressive degeneration and necrosis of the affected cells; karyomegaly with a prominent nucleolus was also a feature of the affected cells. Ultrastructural examination of the affected keratinocytes revealed large and complex, intracytoplasmic virions with elliptical to discoid profiles and an average size of 548.5 nm (SD = 100.58) by 205.65 nm (SD = 21.19) consistent with a typical poxvirus morphology. Furthermore, metagenomic analysis revealed distinct genomic features, whereas phylogenetic analysis of nine core poxvirus genes demonstrated that the seahorse poxvirus forms a unique branch within the teleost poxvirus clade supporting its classification as a novel poxvirus with a proposed species name of Syngnathid cutaneouspox virus. Finally, this study confirms the utility of next-generation sequencing as a methodology for obtaining phylogenetically useful genomic sequences from archival formalin-fixed paraffin-embedded tissues.