Abstract: Micronutrient status is recognized to influence host susceptibility to viral infections, yet its impact on Zika virus (ZIKV) pathogenesis remains incompletely understood. We investigated the effects of dietary selenium and combined selenium plus vitamin E deficiency on ZIKV infection outcomes in a murine model. Mice maintained on deficient diets exhibited significantly lower neutralizing antibody titers and reduced levels of key antiviral cytokines (IFN-γ, TNF-α, IFN-α, IFN-β, IL-12p70, CCL5) compared to controls. Correspondingly, higher viral RNA loads were detected in the brains of double-deficient mice, which also experienced greater weight loss and increased mortality. Deep sequencing revealed no major differences in overall viral genome diversity across diet groups; however, specific mutations, including V330L and D67E in the E gene, and V360I in the NS3 gene, were enriched or detected in nutritionally deficient animals. These findings suggest that antioxidant micronutrient deficiency impairs both humoral and cellular immune responses to ZIKV potentially facilitating enhanced neuroinvasion. While the functional consequences of the identified mutations warrant further investigation, our results underscore the importance of adequate micronutrient intake for optimal antiviral defense. Further studies are needed to clarify the epidemiological significance of these observations.

Abstract: Influenza A viruses exhibit broad host tropism, infecting multiple species including humans, avian species, and swine. Swine influenza virus (SIV), while primarily circulating in porcine populations, demonstrates zoonotic potential with sporadic human infections. In this investigation, we collected pulmonary specimens from infected swine across various regions of Shandong Province, China. Through systematic surveillance, we identified two H1N1 subtype swine influenza A virus strains designated A/swine/China/SD6591/2019(H1N1) (abbreviated SD6591) and A/swine/China/SD6592/2019(H1N1) (abbreviated SD6592). The GenBank accession numbers of the SD6591 viral gene segments are PV464931-PV464938, and the GenBank accession numbers corresponding to each of the eight SD6592 viral gene segments are PV464939-PV464946. Phylogenetic and recombination analyses suggest potential evolutionary differences between the isolates. SD6591 displayed a unique triple-reassortant genotype: comparative nucleotide homology assessments demonstrated that the PB2, PB1, NP, NA, HA, and NEP genes shared highest similarity with classical swine-origin H1N1 viruses. In contrast, SD6592 maintained genomic conservation with previously characterized H1N1 swine strains, though both isolates exhibited varying degrees of intra-segment recombination events. Through comprehensive sequence analysis of these H1N1 SIVs, this study provides preliminary insights into their evolutionary history and underscores the persistent risk of cross-species transmission at the human-swine interface. These findings establish an essential foundation for enhancing national SIV surveillance programs and informing evidence-based prevention strategies against emerging influenza threats.

Abstract: Dengue remains one of the most significant tropical diseases, with its geographic range continuing to expand. The etiological agent of the disease is dengue virus, a term that groups four closely related viruses defined as serotypes. The four dengue virus serotypes elicit distinct serological responses, and their replacement or co-circulation in endemic regions can result in major outbreaks. The identification of the circulating serotypes is essential for surveillance and clinical management. Here we describe a novel method for the detection and serotyping of the four dengue virus serotypes using a single plex multicolor real-time RT-qPCR. The method takes advantage of a highly informative dataset that includes a realistic representation of globally circulating strains, allowing the selection of a universal primer pair and four serotype-specific hydrolysis probes. The method has been extensively tested and the results support its use in surveillance studies as well as in clinical settings.

Abstract: Multiple sclerosis is an immune-driven neurological disease that affect myelinated axons in the central nervous system. However, the trigger of the (dysregulated) immune reactions is not56 known. According to Wilkin’s primary lesion theory, myelin-reactive T cells present in the immune repertoire respond to myelin antigens that are released from idiopathic lesions within the central nervous system. However, neither the cause of the primary lesion, nor the cause of the immune hyper-reactivity are known. We investigated whether these unknown activation signals may be relayed by common herpesviruses. The results highlight human herpesvirus-6A as a potential trigger of primary lesions due to its proven capacity to cause oligodendrogliopathy, cytomegalovirus as a trigger for the formation of effector memory cytotoxic T cells with proven capacity to induce multiple sclerosis pathology in a non-human primate MS model and Epstein Barr Virus due to its capacity to render B cells capable to effectively present a critical myelin antigen to these effector memory cytotoxic T cells. These results lead us to propose the novel paradigm that the immunopathogenesis of multiple sclerosis results from a conspiracy of common herpesviruses.

Abstract: Epidemiological studies have proven that coxsackievirus B3 (CVB3) is the major virus that cause acute and chronic viral myocarditis and cardiomyopathy. Currently, there are no antiviral therapeutic drugs or vaccines to be used as clinical therapeutic or vaccine. Virus-like particles (VLP) based vaccines offer a safe and effective vaccine platforms because of their structural similarity to native viruses, but lack viral genetic material and therefore considered a better immunogens. The viral capsid protein VP1 of CVB3 is the most immunogen viral peptide, providing opportunities for its use in designing VLP-based vaccines or immunodiagnostic reagents. In the present study, we developed and characterized a CVB3 vaccine candidate based on a self-assembly of the major immunogenic viral protein VP1 of CVB3 wild type strain. We assessed its induced humoral and cellular immune responses and then we evaluated its protective immunity against pathogenic CVB3 strain challenges in Balb/c mice model. Neutralizing specific antibodies and Interferon gamma (INF-γ) production were determined in sera of both prime and prime-boosted immunized mice with VLP vaccine candidate. Our results demonstrated that VLP generated by VP1 self-assembled and expressed in eukaryotic insect cell baculovirus vector system elicited potent humoral and cellular immune responses, protecting Balb/c mice from lethal challenges. Hence, the produced VLP based on the self-assembly of VP1 is a promising and potential vaccine candidate against CVB3 natural infections.

Abstract: The 2022 global mpox outbreak represented a turning point in the Monkeypox virus (MPXV) epidemiology, highlighting the incredible capability of this virus to adapt to different conditions also in a non-endemic context. To investigate the genomic dynamics of MPXV 2022 strains, we performed whole-genome sequencing of 40 clinical samples from 16 Italian patients across multiple anatomical sites and timepoints between May and December 2022. Combining single-nucleotide analysis with detailed investigation of short tandem repeats (STRs), we explored inter- and intra-host viral dynamics. We identified 19 STR loci located near or within genes involved in immune modulation and virion morphogenesis. While most STRs remained stable across patients, a subset displayed locus- or matrix-specific variation. Among these, STR-VII—embedded within the coding sequence of OPG153, an envelope-associated protein implicated in viral attachment—showed a 12-nucleotide in-frame deletion, resulting in the loss of four aspartic acid residues (Δ4D variant). Structural modeling indicated that this deletion slightly alters a disordered acidic loop without affecting the global fold, potentially modulating surface charge and immune recognition. Integrating STR profiling into genomic surveillance may enhance resolution in outbreak reconstruction and reveal subtle adaptive processes underlying poxvirus host interaction and immune escape.

Abstract: Despite progress in controlling microbial infections, the global burden of pathogenic bacteria remains high. Moreover, the spread of antibiotic-resistant bacteria has increased and is likely to continue. Consequently, recent research has focused on antibiotic alternatives to slow the proliferation of resistance and improve patient outcomes. One promising approach is phage therapy, which uses lytic viruses of bacteria to treat bacterial infections. However, an underexplored potential benefit is its capacity to enhance health outcomes across the socioeconomic spectrum and improve healthcare equity. Drawing on the history of phage therapy, recent clinical successes, and advancements in research, we argue that phage therapy is well-suited to advance health and healthcare equity. This is because it offers a comparatively greater potential to increase access and reduce disadvantages faced by lower socioeconomic groups relative to conventional antibiotics. Based on these points, we propose research objectives that aim to achieve these scientific and ethical goals.

Abstract: Chikungunya virus (CHIKV) is a mosquito-borne alphavirus prevalent in more than 110 countries and regions, including Africa, Asia, America and Europe. It can cause acute fever, rash and severe joint pain, and some patients may develop chronic arthritis, which significantly impairs the quality of life. Following its resurgence in 2005, CHIKV has emerged as a major threat to global public health. This review summarizes the diagnostic techniques, advances in vaccine development, and the newest drug interventions for CHIKV. We also present an overview of the epidemiology, structure and invasion mechanism of the 2024-2025 epidemic hotspots and propose evidence-based strategies leading to the effective prevention and control of CHIKV.

Abstract: Bloodsucking dipterans are major vectors responsible for the transmission of arboviruses. Additionally, they also carry a lot of insect-specific viruses. High-throughput sequencing technologies has facilitated breakthrough in the study of viruses overall and viruses of haematophagus insects in particular. In this study we used high-throughput sequencing to describe the viromes of bloodsucking dipterans collected in Karelia, northwestern Russia. In addition to various species of Aedes mosquitoes (Diptera: Culicidae), several pool of biting midges (Ceratopogonidae: Culicoides), and blackflies (Simuliidae: Simulium), were analyzed. We managed to assemble and annotate 63 different distinct viruses with complete and partial genomes. The most common viral groups were Durnavirales, Picornavirales, Reovirales, and Mononegavirales, however various other groups of RNA viruses and even one DNA virus were detected. The majority of viruses (78%) were novel viruses. Among known viruses, no known human or animal pathogens were found. Some of the novel viruses can be considered by the International Committee on Taxonomy of Viruses for the demarcation into novel genera.

Abstract: Metapneumoviruses comprise a genus of negative-sense RNA viruses that cause significant respiratory disease across human and avian hosts. Human metapneumovirus (hMPV) is a globally prevalent pathogen associated with acute lower respiratory tract infections in infants, older adults, and immunocompromised individuals, whereas avian metapneumovirus (aMPV) imposes substantial economic losses on the poultry industry through respiratory disease, reproductive impairment, and high mortality in the presence of secondary infections. Despite their distinctive host ranges, hMPV and aMPV share a conserved genomic architecture and encode homologous structural and non-structural proteins that mediate viral entry, replication, assembly, and evasion of host innate im-munity. Comparative analysis highlights both have deeply conserved polymerase and nucleocapsid functions, and yet have a wide range of diversity in the attachment glycoprotein (G) and small hydrophobic protein (SH), reflecting divergent evolutionary pressures in human verses avian hosts that have led to such distinctive differences. The recent emergence and detection of aMPV/A and aMPV/B across the previously aMPV free United States beginning in late 2023, combined with rising cases globally of hMPV post SARS-CoV-2 pandemic, underscore the continued challenges of metapneumovirus surveillance and control in humans and animals. This review aims to highlight the current knowledge on the history, molecular virology, pathogenesis, epidemiology, diagnostics, and control strategies for aMPV while drawing mechanistic parallels to hMPV. By con-textualizing shared biology and structure alongside host-specific adaptations, we aim to identify key gaps that shape vaccine design, antiviral development, and future research priorities aimed at mitigating the health and economic burden posed by metapneumoviruses found in both birds and humans.

Abstract: Gammaretroviruses are ubiquitous pathogens, often associated with the induction of neoplasia, especially leukemia, lymphoma, and sarcoma, and with a propensity to target the germline. The latter trait has left extensive evidence of their infectious competence in vertebrate genomes, the human genome being no exception. Despite the continuing activity of gammaretroviruses in mammals, including Old World monkeys, apes and gibbons, humans have apparently evaded novel infections by the virus class for the past 30 million years or so. Nevertheless, from the 1970’s onward, cell culture studies repeatedly discovered gammaretroviral components and/or virus replication in human samples. The last novel ‘human’ gammaretrovirus, identified in prostate cancer tissue, culminated in the XMRV frenzy of the 2000’s. In the end, that discovery was shown to be due to lab contamination with a murine gammaretrovirus. Contamination is also the likely source of the earlier findings. Complementation between genes of partially defect endogenous proviruses could have been another source of the virions observed. However, the capacity of many gammaretroviruses to replicate in human cell lines, as well as the presence of diverse infectious gammaretroviral species in our animal companions, for instance in mice, cats, pigs, monkeys, chickens, and bats, does not make a transmission to humans an improbable scenario. This review will summarize evidence for, or the lack of, gammaretrovirus infections in humans in the past, the present and the near future. Aspects linked to the probabilities of novel gammaretrovirus infections in humans, regarding exposure risk in connection to –modern– lifestyle, geography, diet, and habitat together with genetic and immune factors, will also be part of the review, as will be the estimated consequences of such novel infections.

Abstract: Liquid-elastic phase separation (LLP) has become an important concept to understand how biomolecular condensate is performed in eukaryote cells, providing a unique opportunity for the virus to take advantage of this system during infections. There is an increasing collection of evidence that suggests that LLPs play a role in almost every stage of the viral life cycle, from early entrance and replication to genome packaging, immunoque and final release. This review collects recent findings of how different viruses utilize LLP to create special, membranous spaces that collect viral and host components, replicate replication efficiency and avoid the host defense. It also engages in potential therapeutic strategies aimed to target viral LLP, such as disrupting scaffolding protein interactions, changing the properties of condensate and disrupting changes after translation. While these approaches reflect promises, they face obstacles related to the unique, distribution and complex role of LLP in both hosts immune. Getting a deeper understanding of molecular complications of viral LLP, it would be important to develop innovative antiviral strategies, such as the functions of the internal and RNA structure, internal and the RNA structure. As the research develops, LLPS provides a new structure form to understand viral pathogenesis and drug discovery.

Abstract: HIV-1 entry into host cells culminates in integration of the reverse transcribed double-stranded viral DNA into host genes. Several lines of evidence suggest that intact, or nearly intact, HIV-1 cores – large ~60 nm-wide structures – pass through the nuclear pore complex (NPC) and that this passage is associated with pore remodeling. Cryo-electron tomography studies support the dynamic nature of NPCs and their regulation by cytoskeleton and ATP-dependent processes. To explore NPC remodeling, we used super-resolution Stochastic Optical Reconstruction Microscopy (STORM) of U2OS cells endogenously expressing nucleoporin 96 tagged with SNAP. Single molecule localization imaging and computational averaging resolved 8-fold symmetric nuclear pores with an average radius of ~51 nm. Depletion of cellular ATP using sodium azide or antimycin A, previously reported to reduce the size of yeast NPCs, did not significantly alter the nuclear pore radius in U2OS cells. Similarly, stressing the nuclear envelope by hypotonic or hypertonic conditions failed to induce detectable expansion/contraction of NPCs. These results indicate that the NPCs in U2OS cells do not respond to ATP depletion nor mechanical stresses on changes in pore morphology that can be resolved by STORM. Since these cells are infectable by HIV-1, we surmise that direct multivalent interactions between HIV-1 capsid and phenylalanine-glycine nucleoporins lining the pore’s interior drive the core penetration into the nucleus and the associated changes in the pore structure.

Abstract: Infectious Bronchitis Virus is one of several major viral infections in poultry, affecting the respiratory, reproductive, and renal systems, and causing significant economic losses worldwide. Current vaccines, including the H120 strain, provide limited cross-protection against emerging variants, underscoring the need for improved vaccine strategies. In this study, the complete genome of IBV H120 was divided into 12 fragments, synthesized, and assembled using the Golden Gate Assembly (GGA) method. The recombinant virus (rH120) was successfully rescued in chicken fibroblast cells and propagated in embryonated chicken eggs. Growth kinetics in embryonated chicken eggs revealed similar replication patterns between rH120 and the original H120 strain. In broiler chickens, rH120 replicated efficiently as confirmed by viral RNA detection in throat and cloacal swabs, and induced a stronger antibody response by 14 days post-infection. The rH120 virus proved to be genetically stable, infectious, and immunogenic, indicating that GGA-based reverse genetics is an effective system for IBV vaccine development.

Abstract: Mpox virus lineages associated with sustained human-to-human transmission, Clade Ib/sh2023 and Ia/sh2024, are co-circulating in Kinshasa, Democratic Republic of the Congo (DRC). We report the first two Clade IIb/sh2017 cases identified in DRC, imported from West Africa and locally transmitted, with phylogenetic linkage to the ongoing outbreak in Sierra Leone.

Abstract: The potyvirus helper component proteinase (HcPro) is a multifunctional protein, one of its most documented functions being host antiviral RNA silencing suppression. This study shows that the HcPro of potato virus Y (PVY), an important member of the potyvirus group, prevents replication of a related competing secondary virus. This phenomenon, referred to as superinfection exclusion (SIE), is common in bacterial, human, and plant virus infections. We also report that HcPro induction of SIE is strain specific and this specificity is provided by the first four amino acid residues of the protein. Consistent with the mechanism of SIE, the study found that HcPro does not exclude a resident virus. Additionally, HcPro induction of SIE was observed to function independently of its ability to suppress antiviral RNA silencing. HcPro induction of SIE is relevant given the prevalence of multiple PVY strains that routinely co-infect the same cell and that may lead to recombination and emergence of new and more virulent strains. Furthermore, cross-protection that is employed in plant virus disease management occurs when SIE moves from the cellular level and spreads systemically, emphasizing the importance of SIE.

Abstract: Porcine cytomegalovirus, more accurately classified as porcine roseolovirus (PCMV/PRV), was shown pathogenic in the context of xenotransplantation. Transmission of PCMV/PRV to non-human primates receiving hearts or kidneys from virus-positive pigs significantly reduced the survival time of the recipients. PCMV/PRV was also transmitted to the first human recipient of a pig heart transplant and contributed to the patient’s death. PCMV/PRV is highly prevalent in all pig breeds and wild boars, including slaughterhouse pigs, but there were no reports of infection or disease in either healthy, ill or immunocompromised humans. This indicates that this virus is not zoonotic and must be classified as xenozoonotic. Moreover, it remains unclear whether PCMV/PRV is capable of infecting human cells in vitro. To address this question, human 293T cells resistant to hygromycin were co-cultured with porcine fallopian tube (PFT) cells producing PCMV/PRV. After hygromycin selection, the remaining human cells showed no evidence of infection. Because herpesviruses are generally considered to be species-specific—a notion that has been shown to be not entirely correct—it was also investigated whether PCMV/PRV can infect mouse cells using the same approach. Similarly, no infection was observed. Since the target cells employed in both assays had a reduced capacity to resist viral infection, the findings strongly suggest that PCMV/PRV is unable to infect human or mouse cells, which are equipped with functional antiviral mechanisms.

Abstract:

Parvovirus B19 and cytomegalovirus are significant causes of congenital infections that can lead to adverse pregnancy outcomes. The present study aimed to assess the prevalence of B19V and CMV infections in pregnant women with fetal anemia and/or effusions and determine the utility of routine laboratory screening in pregnancy follow-up. Thirteen women with such pathological pregnancy complications attending an antenatal clinic from April 2024 to March 2025 were tested. Four types of clinical material were examined: amniotic fluid, umbilical cord serum, maternal blood, and fetal blood. Participants underwent molecular and serological testing for both B19V and CMV. Demographic data, obstetric histories, and pregnancy outcomes were recorded and analyzed. Our results indicate that four participants showed evidence of either current infection with CMV and seven with B19V. Pregnant women with active infections required further follow-up and fetal surveillance. A stillbirth was reported in one woman with CMV infection. For seven samples that tested positive for B19V DNA, viral sequences were obtained and clustered with genotype 1a reference strains. Implementing routine or targeted screening for B19V and CMV could facilitate the early identification and appropriate management of maternal infections, ultimately reducing the risk of fetal complications.

Abstract: Background: Seasonal influenza remains a significant public health problem, and the constant antigenic drift of viruses requires regular vaccine updates. mRNA vaccines offer a promising platform for the development of new, effective influenza vaccines. Administration of the naked mRNA vaccine using a needle-free jet injection system further enhances its safety, reduces cost, and eliminates the need for lipid nanoparticles, which are traditionally used for mRNA delivery. Lyophilization of naked mRNA allows for longterm storage at +4°C. Methods: We designed and produced an mRNA vaccine against seasonal influenza, designated mRNA-Vector-Flu, encoding the hemagglutinin (HA) of the A/Wisconsin/67/2022(H1N1)pdm09, A/Darwin/9/2021(H3N2), and B/Austria/1359417/2021 strains. The vaccine was lyophilized and stored for 1 month in a refrigerator (+4°C). A comparative immunogenicity study was conducted between synthesized immediately before use prepared and lyophilized naked mRNA-Vector-Flu. The preparations were administered to BALB/c mice using a jet needleless injection twice, 3 weeks apart. Immunogenicity was assessed on day 35 of the study. Results: A comparative immunogenicity study of naked mRNA-Vector-Flu demonstrated that both the synthesized immediately before use prepared formulation and the lyophilized form, stored at +4°C for a month, induced similar levels of virus-specific antibodies and generated a pro-nounced T-cell immune response. Conclusions: Delivery of the naked mRNA vaccine us-ing a needle-free jet injection ensures a high-level immune response, which improves its safety, reduces its cost, and eliminates the need for lipid nanoparticles traditionally used for mRNA delivery. At the same time, lyophilization of the naked mRNA vaccine pre-serves its biological activity and ensures its long-term storage at above-zero temperatures. Our results demonstrate that our proposed approach can be considered a promising di-rection for the development and improvement of the mRNA vaccine platform.

Abstract: Human cytomegalovirus (HCMV) has emerged as a potential oncomodulatory agent implicated in the pathogenesis of several cancers, although its precise role in tumorigenesis remains a subject of debate. This review synthesizes five decades of research examining the involvement of HCMV in cancer, with particular focus on molecular mechanisms by which the virus may contribute to tumor progression, including immune evasion, chronic inflammation, and modulation of cell cycle and signaling pathways. Evidence of HCMV detection in malignancies such as glioblastoma, breast, colorectal, and prostate cancers is critically reviewed, alongside ongoing controversies regarding its causal versus passenger role in oncogenesis.The review further explores current and emerging antiviral and immunotherapeutic strategies targeting HCMV within oncological settings, highlighting their potential translational relevance and associated challenges. By reassessing oncogenic hypotheses and therapeutic opportunities, this article aims to provide a comprehensive perspective on the complex interplay between HCMV and cancer biology, and to evaluate its potential as a novel target for cancer prevention and therapy.