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Archives of Virology

Springer Science and Business Media LLC

Preprints posted in the last 30 days, ranked by how well they match Archives of Virology's content profile, based on 15 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

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Isolation and characterisation of novel fruit bat alphaherpesvirus from Rousettus aegyptiacus bats in Coastal Kenya

Kisoi, G. K.; Bargul, J.; Kinyua, J.; Langat, S.; Koka, H.; Lutomiah, J.; Eyase, F.

2026-06-25 microbiology 10.64898/2026.06.25.734443 medRxiv
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BackgroundHerpesviruses are a group of double-stranded DNA viruses known to infect a wide range of vertebrates and establish life-long latent infections. While bats serve as natural reservoir hosts for numerous viral families, relatively few bat herpesviruses have been successfully isolated. In this study, we report the isolation and characterization of two novel alphaherpesvirus strains obtained from Rousettus aegyptiacus bats in Coastal Kenya. MethodsThe samples of oral and rectal swabs were collected from three different species of bats from coastal Kenya between October 2024 and April 2025; the bat species collected include Hipposideros spp., Coleura afra, and Rousettus aegyptiacus. Virus isolation was performed by inoculation of samples in Vero E6 cells and subsequent monitoring for cytopathic effects (CPE). Total nucleic acids were extracted from CPE positive cultures and subjected to library preparation to enable unbiased detection of both RNA and DNA viruses. The libraries were sequenced using next-generation sequencing with Illumina MiSeq platform. Subsequently, bioinformatic analysis was carried out to identify the virus, generate consensus genomes as well as phylogenetic analysis to determine the placement of identified viruses. ResultsTwo samples from R. aegyptiacus (KIK_460_O and KIK_465_O) induced typical CPE within five days. Sequencing and assembly yielded partial consensus sequences of approximately 60 kb (KIK_460_O) and 70 kb (KIK_465_O), representing extended genomic data for a bat-associated alphaherpesvirus. This virus has a genome of about 140kb, indicating that our partial assemblies account for about 43-50% of the total genome. Both isolates were found to be closely related to Dzifa herpesvirus, an alphaherpesvirus previously identified in Kilifi, Kenya. Alphaherpesvirus was identified based on partial sequencing of UL19 (3,787bp) and UL30 (2,846bp) genes. The two isolates were found to be identical at the UL19 gene, showing that they belonged to the same virus strain. Phylogenetic analysis showed that the novel alphaherpesvirus belongs to primate alphaherpesviruses under the subfamily Alphaherpesvirinae. ConclusionThis study reports the isolation and genomic characterization of a novel fruit bat alphaherpesvirus from Kenyan Rousettus aegyptiacus bats. The partial genome assembly (60-70 kb) represent the first extended genomic data for this virus, covering approximately 43-50% of the estimated 140 kb complete genome. The phylogenetic placement of this alphaherpesvirus near primate viruses, especially Pteropodid alphaherpesvirus 1, suggests bat-association and needs further investigation into its zoonotic potential.

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Characterization of an Arctic-like 1a rabies virus from a 54-day-old puppy with atypical presentation, Pune, India, 2026

Ullas, P. T.; Sharma, V.; Vipat, V.; Choudhari, S.; Ashraf, A. F.; Raju, R. M.; Kotturi, V.; Sakhare, K. S.; Bondre, V. P.

2026-07-13 infectious diseases 10.64898/2026.07.09.26357633 medRxiv
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Rabies remains a significantly underreported fatal zoonosis in India, where the Arctic-like 1a (AL1a) lineage predominates in dog populations. While atypical clinical presentations in dogs can delay diagnosis and increase human exposure risk, genomic and clinical data on neonatal canine rabies remain limited. This study reports an exceptional case of rabies in a 54-day old unvaccinated German shepherd puppy which presented with severe pruritus and self-biting behaviour. The puppy was euthanized due to poor clinical response. Post-mortem testing revealed viral antigen (by Direct Fluorescent Antibody Test) and viral RNA (by real-time RTPCR) in the brain tissue. Whole-genome sequencing recovered a near-complete rabies virus genome (11,947 nucleotides; 99.5% genome coverage), classified within the AL1a_A1.1 sublineage. Phylogenetic analysis revealed close genetic relatedness to contemporary Indian rabies virus strains. Comparative genomic analysis identified 4, 3, 6, and 8 non-synonymous substitutions in the phosphoprotein, matrix, glycoprotein, and polymerase genes, respectively. This case is one of the youngest documented cases of canine rabies with atypical manifestations, caused by the AL1a viral clade. Our findings highlight the risks associated with neonatal canine rabies, the need for heightened clinical suspicion in atypical cases, and the importance of genomic surveillance to monitor evolving rabies virus lineages in endemic regions.

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Genomes of Betacoronavirus gravedinis from white-footed mice in New York City and a phylogenetically weighted model of its probable distribution in North America

Kaza, B.; Catchen, M.; de Gennaro, G.; Zehr, J.; Lilly, M.; Plimpton, L.; Diuk-Wasser, M.; Murrell, C.; Ishee, A.; Goodman, L.; Whittaker, G.; Gamble, A.; Olarte-Castillo, X.

2026-07-01 microbiology 10.64898/2026.06.30.735598 medRxiv
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Rodents are an important reservoir of zoonotic viruses and are ubiquitously present in densely populated urban areas. Betacoronaviruses in the Embecovirus lineage are well known to infect both humans and animals and have established rodent reservoirs. Here three Betacoronavirus gravedinis genomes were sequenced and characterized in white footed mice (Peromyscus leucopus, commonly white footed mice) collected in New York City, the second most populous city in North America. The genomes were distinct from mouse hepatitis virus (MHV), the prototype mouse betacoronavirus, and highly similar and identical in one case to previously characterized B. gravedinis sequences from white footed mice in Connecticut. Codon aware evolutionary models were used to identify specific sites under positive selection within the spike protein of B. gravedinis. A novel method was developed to predict the probable geographic distribution of the virus using publicly available data from the Global Biodiversity Information Facility to generate a weighted distribution map highlighting overlapping potential host ranges based on the evolutionary distance using a high resolution cytocrome B (CYTB) phylogeny of rodent species with potentially overlapping ranges. Our models predict three current hotspots of circulation in North America under different possible transmission regimes, and an additional fourth hotspot was predicted to arise in a warming future. This study highlights the continued need for biodiversity-informed surveillance of potential zoonotic pathogens in rodents.

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Emergence and co-circulation of Monkeypox virus Clade Ia and Clade Ib in South Kivu, Democratic Republic of the Congo, January to May 2026

Murhula, L.; Udahemuka, J.; Nieuwenhuijse, D. F.; Chasinga, B.; Sindayiheba, R.; Schuele, L.; Cassidy, H.; Bacon Benimana, F.; Chigabo, A.; Bihando, J.; Nzigire Barhatwira, G.; Bengehya Mbiribindi, J.; Ndoli Minega, J.; Lang, T. A.; Lulihoshi Willy, K.; Ngabo, P.; Mitchell, S.; Gortazar, C.; M. Aarestrup, F.; Bahizire, E.; Koopmans, M.; Oude Munnink, B.; Ndishimye, P.

2026-06-29 epidemiology 10.64898/2026.06.25.26356562 medRxiv
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In September 2023, the first infections with a novel lineage of mpox were detected in South Kivu. Since then, the virus has spread regionally, nationally and internationally. As part of continued efforts to understand the mpox ecology and epidemiology, the South Kivu district of public health and partners have set up systematic case finding and follow-up, including strain characterisation through PCR and sequencing. Samples were collected from 595 hospitalized patients with a confirmed mpox virus infection. A clade differentiating RT-PCR showed that 545 (92%) of samples were positive for clade Ib but also remarkably that Clade Ia infections were diagnosed for the first time in South Kivu. First detected in cases in week 7 in Kamituga, 50 cases were identified over the whole study period (8,40% of all cases). Phylogenetic analysis of initial cases revealed introductions of clade Ia into the South Kivu province alongside the continuation of the clade Ib mpox virus outbreak. These findings underscore the increasing complexity of clade I mpox virus outbreaks in the DRC.

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Isolation of Zika Virus Replication Complex Reveals a Proviral Nuclear Factor

Chang, P.; Sallapalli, B. T.; Zhang, Y.-J.

2026-07-07 microbiology 10.64898/2026.07.06.736844 medRxiv
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Zika virus (ZIKV) is an arthropod-borne flavivirus of international public health impact. ZIKV has a positive-sense, single-stranded RNA genome and remodels intracellular membranes to form replication complexes (RCs). The objective of this study was to isolate and characterize the RCs from ZIKV-infected cells and to identify host-cell components recruited to participate in viral replication. Here, we isolated the RCs from ZIKV-infected Vero cells by detergent treatment and flotation centrifugation. Fractional flotation analysis demonstrated that ZIKV proteins NS2B, NS3, and NS5, and ZIKV RNA were present in the detergent-resistant membranous fraction. In contrast, the ER-resident protein calnexin and a mitochondrial protein were present in the detergent-soluble fractions. The isolated RCs were functional for ZIKV RNA synthesis, as shown by quantitative PCR. To determine the components of the RCs, we conducted mass spectrometry analysis and identified numerous cellular proteins. Among them is the replication factor C subunit 2 (RFC2), an accessory protein of DNA polymerase. RFC2 is involved in ATP binding and hydrolysis and may promote cell survival. ZIKV infection increased the RFC2 protein level and induced its relocation to the cytoplasm. RNAi-mediated silencing of RFC2 reduced ZIKV replication. Together, our results provide insights into ZIKV replication and virus-cell interaction.

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Effect of CSFV on Differential Genes of Histone Lactylation at H3K18 in the PI3K-AKT Signaling Pathway

Zhang, H.; Han, Z.; Zhao, X.; Zhu, J.; Shao, N.; Sun, K.; Li, W.; Yao, Y.; Liang, X.; Yang, M.; Gao, Y.; Chen, J.; Liang, Y.; Liu, Q.; Li, X.; Cao, Z.

2026-06-29 microbiology 10.64898/2026.06.26.734696 medRxiv
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Classical swine fever (CSF) is a highly contagious disease caused by Classical swine fever virus (CSFV), posing a serious threat to the global swine industry. This study aimed to investigate the effect of CSFV on differential genes of histone lactylation at the H3K18 site in the PI3K-AKT signaling pathway. The site with the most significant change in histone lactylation antibody level was screened by Western blot. Omics analysis was performed using CUT&Tag technology to identify differential genes in the PI3K-AKT pathway between the CSFV-infected group and the mock group, followed by validation using RT-qPCR. Functional analysis of significantly differential proteins was conducted, and the protein expression level of THBS4 was detected by Western blot. The results showed that after CSFV infection of 3D4/21 cells, the H3K18la site exhibited the most significant difference in antibody level. A total of 8,859 differential genes at the H3K18la site were identified by CUT&Tag analysis, including 6,349 up-regulated genes and 2,510 down-regulated genes. Further focusing on the PI3K-AKT signaling pathway, 10 differential genes were identified, comprising 6 up-regulated genes and 4 down-regulated genes. Compared with the control group, the mRNA expression levels of CD19, LAMA1, PDGFRA, BDNF, ANGPT4, and THBS4 were up-regulated in the CSFV-infected group, while FOXO3 and NRTN were down-regulated. Western blot results showed that the protein expression level of THBS4 increased after CSFV infection. These findings lay an important foundation for understanding the molecular mechanisms regulating viral replication and immune evasion, and have significant scientific implications and potential application value.

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Interaction of Bunyamwera Virus Non-Structural Protein NSm with Cellular BNIP1 is Required for Efficient Viral Gene Expression and Replication

Wartnaby, R. F.; Fontana, J.; Barr, J. N.

2026-07-01 microbiology 10.64898/2026.07.01.735799 medRxiv
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Bunyamwera virus (BUNV) is the prototypical member of the Peribunyaviridae family of arthropod-borne viruses and possesses a genome comprising three segments of negative-sense RNA, named small, medium and large. The medium segment encodes a polyprotein that is processed to form Gn and Gc spikes and a non-structural protein, NSm. The role of NSm during replication in mammalian cells is poorly characterized, although it associates with a Golgi-derived structure called the virus factory (VF), the site of BUNV genome replication and virion assembly. To further define NSm function, we generated an epitope-tagged BUNV and used co-immunoprecipitation and quantitative proteomics to identify host interacting partners. NSm interacted with BCL-2 interacting protein 1 (BNIP1), a SNARE protein involved in COPI vesicle trafficking, with the importance of this interaction demonstrated by siRNA-mediated knockdown of BNIP1 expression, which significantly reduced BUNV gene expression and virion production. Interestingly, NSm also interacted with components of the NRZ complex, involved in COPI vesicle tethering in association with BNIP1, and inhibition of COPI complex formation resulted in loss of NSm expression. Taken together, our results identify BNIP1 as a host cell factor necessary for efficient BUNV replication and suggest the cellular localization of NSm at the VF is COPI-dependent.

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Shifts in Genetic Diversity of Porcine Reproductive and Respiratory Syndrome Virus 2 in Vietnam Before and After African Swine Fever: Increased Diversity and Novel Sub-lineages

Nguyen, T. C.; Pamornchainavakul, N.; Herrera da Silva, J. P.; Thanawongnuwech, R.; VanderWaal, K.

2026-07-10 genetics 10.64898/2026.07.06.736905 medRxiv
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Porcine reproductive and respiratory syndrome virus 2 (PRRSV-2) remains one of the most important transboundary pathogens affecting swine production in Vietnam; however, it remains poorly understood how long-term evolutionary dynamics were impacted by the African swine fever (ASF) epidemic, a period of time where swine population demographics and movement were heavily perturbed. We investigated the molecular epidemiology, evolutionary history, and phylogeographic dynamics of PRRSV-2 circulating in Vietnam between 2007 and 2024 by integrating 366 Vietnamese ORF5 sequences with a globally curated lineage reference. Maximum-likelihood phylogenetic, Bayesian phylodynamic, and discrete phylogeographic analyses revealed that the Vietnamese PRRSV-2 population underwent substantial reshaping after the ASF epidemic, shifting from a predominantly endemic sub-lineage L8E population to a genetically diverse viral community comprising multiple established and newly emerging sub-lineages. Despite these epidemiological changes, the endemic sub-lineage L8E population maintained a relatively stable evolutionary rate across the pre- and post-ASF periods, suggesting that ASF reshaped viral population structure rather than intrinsic evolutionary dynamics. Two previously unclassified viral clusters circulating in Vietnam and Thailand fulfilled all criteria for formal designation and were recognized as the novel sub-lineages L1M and L10B by the International PRRSV-2 Nomenclature Consortium. Phylogeographic reconstruction further demonstrated contrasting transmission patterns among major sub-lineages, including long-term endemic persistence of L8E, repeated unidirectional introductions of sub-lineages L1M and L10B from Thailand, and bidirectional transpacific dissemination of sub-lineage L1A linking Southeast Asia and North America. Collectively, these findings demonstrate that the ASF epidemic coincided with a fundamental reshaping of the PRRSV-2 epidemiological landscape in Vietnam while revealing Southeast Asia as an active center of ongoing viral diversification. This study provides an updated evolutionary framework for PRRSV-2 surveillance and highlights the importance of continuous genomic monitoring and regional collaboration for the early detection and control of emerging transboundary variants.

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Genomic Surveillance of Respiratory Syncytial Virus among Patients with Acute Respiratory Infection through Hospital-Based Influenza Surveillance Platforms in Bangladesh, August 2024-December 2025

Alam, M. S.; Begum, M. N.; Rahman, M.; Chowdhury, F.; Jubair, M.; Karim, Y.; Shanto, M. R. R.; Howlader, R.; Rahman, T.; Talha, M.

2026-07-03 evolutionary biology 10.64898/2026.07.02.736091 medRxiv
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Background: RSV is a major cause of severe lung infections in young children, with over 95% of deaths occurring in poorer countries. Bangladesh has high rates of RSV illness in children but lacks genetic data from after the COVID-19 pandemic. New vaccines and antibody treatments are now available, making local genetic information essential. Objectives: We sequenced complete RSV genomes from Bangladeshi patients to study virus types, genetic changes, and protein mutations, and shared our data openly. Methods: From August 2024 to December 2025, we took 59 RSV-positive samples with high virus levels from hospital patients and sequenced their full genomes using Oxford Nanopore technology. Results: Among 11,874 patients, 1,390 (11.7%) had RSV, mostly RSV-A (94.6%). We obtained 49 good-quality full genomes from the 59 samples (83% success): 43 RSV-A (ON1 type, five sub-lineages) and 6 RSV-B (BA9 type). We found S276N in 35% of RSV-A and S389P in all RSV-B, but neither stops current antibody treatments. All RSV-A viruses gained a new sugar attachment site on their F protein, and most RSV-B viruses gained one too. We uploaded all 49 genomes to GISAID for public use. Conclusion: This work shows we can do full RSV genome sequencing in Bangladesh. The viruses here still match the targets of new vaccines and antibodies, which is reassuring. Our findings provide a foundation for planning RSV prevention in Bangladesh and South Asia.

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Single-Cell Transcriptomic Analysis of the Immune Response to CHIKVInfection

Huang, P.; Wang, H.; Xu, S.; Li, M.; Guo, M.; Wang, H.; Gou, X.; Wang, C.; He, Y.; Pan, W.

2026-06-30 immunology 10.64898/2026.06.24.734421 medRxiv
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Abstract Background: The Chikungunya virus (CHIKV), a re-emerging mosquito-borne alphavirus, is responsible for acute febrile illness and severe polyarthralgia. Although both innate and adaptive immune responses influence the disease outcomes, the detailed cellular immunopathogenesis of CHIKV in peripheral blood is not yet fully elucidated. Methods: We conducted single-cell RNA sequencing (scRNA-seq) on peripheral blood mononuclear cells (PBMCs) obtained from patients acutely infected with CHIKV and from healthy control subjects. Cellular interactions were inferred, and the transcriptomic results were orthogonally validated through quantitative real-time PCR (qPCR) and Enzyme-Linked Immunosorbent Assay (ELISA) to assess systemic interferon-stimulated responses. Furthermore, a comparative analysis was performed using publicly available single-cell data from Dengue virus (DENV) infections. Results: CHIKV infection significantly altered the immune system, increasing monocytes and dendritic cells while reducing T and B lymphocytes. Monocytes and NK cells showed strong activation of interferon-stimulated genes (ISGs). Monocytes were identified as key in driving inflammatory and immune responses. In adaptive immunity, CHIKV led B cells to become plasmablasts with antiviral immunoglobulins and caused T cells and NK-like T cells to show signs of cytotoxicity and exhaustion. Validation showed increased levels of IFN-{gamma}, IFN-{beta}1, MX1, and ISG15. CHIKV triggered a more intense, monocyte-driven interferon response than DENV. Conclusions: Acute CHIKV infection induces a systemic interferon response predominantly centered on monocytes, accompanied by significant alterations in adaptive immunity. Circulating ISG products, including MX1 and ISG15, reflect the transcriptomic activation and may serve as potential biomarkers for assessing the early intensity of innate antiviral responses.

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Genomic descriptive study of human Parvovirus B19 circulation during 2024/2025 in the State of Rio de Janeiro, Brazil

Silveira, M. C.; Azevedo, R. C.; Lamarca, A. P.; Guimaraes, M. A. A. M.; de Mello, C. M. B.; Camargo, A. C.; Gerber, A. L.; Guimaraes, A. P. C.; Cavalcanti, A. C.; Vasconcelos, A. T. R.

2026-07-06 public and global health 10.64898/2026.07.03.26357244 medRxiv
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Purpose: Human Parvovirus B19 (B19V) infection is associated with a broad spectrum of clinical manifestations, including erythema infectiosum, arthropathy, transient red cell aplasia, hepatitis, and adverse fetal outcomes. Due to nonspecific presentations and limited routine testing, B19V infection often remains undiagnosed. Despite recent reports of increased B19V activity worldwide, contemporary data on its presence in Brazil remain scarce. We investigated the circulation of underdiagnosed pathogens in patients with suspected infectious diseases, prioritizing severe and fatal cases. Methods: In this descriptive study, hybrid capture-based whole-genome sequencing was used for a broad-range viral detection and characterization. We analyzed 472 clinical specimens selected for diagnostic investigation between 2024 and 2025, according to surveillance criteria for respiratory and arboviral infections, referred to the Public Health Laboratory of the state of Rio de Janeiro, Brazil. Results: B19V was the second most frequently detected viral pathogens, being identified in 190/472 specimens. Of these, just one sample had been previously tested for this pathogen. Thirty-one cases had higher B19V genomic coverage and were therefore selected for further analysis. Notably, B19V was the only virus detected with substantial genomic coverage in nine individuals, including elderly patients, and meningitis cases with B19V-positive cerebrospinal fluid. Phylogenetic analysis showed that recovered genomes clustered within genotype 1A2. Conclusions: B19V was frequently detected in patients undergoing investigation for infectious diseases and its circulation may represent an important and underrecognized pathogen. These findings support the incorporation of B19V testing into diagnostic algorithms for unexplained infectious diseases, especially in patients presenting atypical symptoms.

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Isolation and characterization of novel filamentous phages from Swiss-type cheeses infecting the Gram-positive bacterium Propionibacterium freudenreichii

Grosset, N.; Nicolas, A.; Jardin, J.; Oechslin, F.; Culot, A.; Moineau, S.; Gautier, M.; GUEDON, E.

2026-07-13 microbiology 10.64898/2026.07.11.737922 medRxiv
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Filamentous phages infecting Gram-positive bacteria remain largely unexplored. Notably, only two filamentous phages, B5 and Philemon infecting Propionibacterium freudenreichii, have been described to date in the phage-rich dairy ecosystem. Although both were genomically characterized, only B5 was confirmed to be an infective filamentous single-stranded DNA phage. The aim of this study was to isolate and characterize new filamentous phages from Swiss-type cheese to investigate their diversity, structural features, host specificity, and potential adaptation to the dairy environment. Thirty raw and pasteurized milk cheeses from France were screened for phages infecting P. freudenreichii strains. Eleven phages were isolated, nine of which displayed a filamentous morphology. Named MINOG1 to MINOG9, these filamentous phages exhibited genomic features typical of this morphotype, including small single-stranded DNA genomes with collinear genes organized into functional modules. Comparison with B5 and Philemon revealed sequence divergence ranging from 0.1% to 7%. These phages also exhibited a diverse host range. To further explore phage-P. freudenreichii interactions, we screened the genomes of the strains used in this study, as well as additional genomes retrieved from the NCBI database, for CRISPR spacers predicted to target these filamentous phages. Numerous strains contained CRISPR spacers showing 79 to 100% identity to genomic regions of these phages. Two P. freudenreichii strains displayed markedly different phage resistance levels despite exact spacer-protospacer matches with phages B5, MINOG1, MINOG2, and MINOG8. Conversely, several strains were resistant to nearly all tested phages despite lacking CRISPR spacers targeting them suggesting the presence of additional defense systems in P. freudenreichii. IMPORTANCEFilamentous phages can play important roles in bacterial ecology by modulating host physiology, population dynamics, and bacterial adaptation to specific environments. However, filamentous phages infecting Gram-positive bacteria remain among the least explored bacterial viruses, and their diversity, ecology, and interactions with their hosts are still poorly understood. This knowledge gap is particularly relevant in dairy ecosystems, where phages are abundant and can influence microbial communities and fermentation processes. In characterizing nine new filamentous phages infecting Propionibacterium freudenreichii from Swiss-type cheeses, this study expands the known diversity of filamentous phages associated with Gram-positive bacteria and provides new insights into phage-host interactions and bacterial defense strategies in dairy-associated bacteria.

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Landscape of non-SARS-CoV-2 respiratory virus sequence data in Africa

Kwok, K.; Mojsiejczuk, L.; Hughes, J.; da Silva Filipe, A.; Ho, A.

2026-07-01 infectious diseases 10.64898/2026.06.27.26356737 medRxiv
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Background Sequencing capacity in Africa greatly expanded during the COVID-19 pandemic. However, the availability of sequence data for non-SARS-CoV-2 respiratory viruses in the region remains uncertain. We systematically analysed sequence data from GenBank, GISAID and Pathoplexus for 12 non-SARS-CoV-2 respiratory virus groups from Africa and compared Africa's genomic record availability with global genomic datasets. We further examined global genomic sequences to identify virus-specific patterns that could inform respiratory virus monitoring efforts in Africa. Results In Africa, the most sequenced virus was respiratory syncytial virus (RSV) (n=15,452), followed by influenza A virus (IAV) (n=10,900) and rhinovirus (n=4,774), when all sequences, including partial ones, were considered. Kenya and South Africa together contributed more than 60% of African respiratory virus sequences, while 30% of African countries submitted none. Within the global dataset of near-complete genomes, IAV was the most sequenced virus in Africa, consistent with global trends. However, Africa contributed the second-lowest number of genomes per million population among all world regions, exceeding only Asia. Substantial differences in relative genetic diversity were observed across viruses. Cytomegalovirus (94%) and parechovirus (38%) showed high sequence retention after clustering, with their clusters typically comprising sequences from a single region. In contrast, influenza viruses and RSV exhibited lower retention, with most clusters circulating across world regions. Additionally, sequences generated through next-generation sequencing were generally more complete than those obtained using Sanger sequencing. Conclusions Our analysis revealed substantial regional disparities in respiratory virus sequence availability across Africa, along with differences in diversity and geographical distribution patterns observed among viruses. This indicates the need for tailored and virus-specific surveillance strategies. Expanding sequencing capacity in more African nations is crucial for a clearer understanding of respiratory viruses circulating in Africa. Ultimately, this can guide vaccine and diagnostic development as well as performance assessment, aiding pandemic preparedness through the timely detection of emerging respiratory viruses.

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Long read and short read whole genome sequencing are equivalent for genomic characterisation of bacteriophage: considerations for high throughput analysis

Carr, P. G.; Iszatt, J. J.; Hedges, M. G.; Mantjani, L.; Vaitekenas, A.; Stick, S. M.; Kicic, A.; Montgomery, S. T.; Phage WA,

2026-07-09 microbiology 10.64898/2026.07.08.737226 medRxiv
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Background: Antimicrobial resistance (AMR) is a global health crisis, necessitating alternative antibacterial strategies. Bacteriophages (phages) offer a promising solution, and their use as a therapeutic agent relies on stringent bioinformatic characterisation using whole genome sequencing (WGS) technologies. However, phages are highly diverse, with no clear consensus on best practices concerning phage DNA extraction or sequencing platform. Efficient and repeatable DNA extraction, sequencing, and bioinformatics processes are critical for safety assessments but remain poorly defined. Additionally, the impact of sequencing platform choice and DNA extraction methods on downstream genomic analyses is not well understood. Methods: We evaluated multiple DNA extraction, library preparation, and sequencing approaches using a diverse collection of Pseudomonas phages from the PhageWA biobank. Column-based and precipitation-based DNA extraction methods were compared for DNA yield and recovery efficiency. Genome sequencing was performed using short-read (Illumina) and long-read (Oxford Nanopore Technologies) platforms, incorporating multiple library preparation kits and Nanopore basecalling models. Assemblies were assessed for completeness, quality, and sequence concordance using standardised bioinformatics pipelines, with hybrid Illumina-Nanopore assemblies used as references for comparison. Results: DNA extraction efficiency varied substantially between protocols, with the Puregene precipitation-based method yielding significantly higher DNA recovery than column-based approaches when normalised to phage titre. Illumina sequencing consistently generated complete genome assemblies, although assembly fragmentation was observed for several jumbo phages when using the SeqWell ExpressPlex 2.0 library preparation method. For Nanopore sequencing, ligation-based native barcoding libraries produced longer reads than rapid barcoding libraries, while selection of the Dorado v5.0.0 basecalling model significantly improved read quality. Genome assembly success was dependent on phage genus; native Nanopore sequencing failed to assemble several Pbunavirus genomes, likely due to modified DNA bases, but an amplification-based library preparation successfully resolved these genomes. Across successfully assembled samples, Illumina and Nanopore platforms produced highly concordant genomes with comparable completeness scores, and hybrid polishing identified only minor sequence differences. Conclusions: DNA extraction methodology, sequencing chemistry, and basecalling model selection significantly influence phage WGS outcomes. Precipitation-based DNA extraction improved DNA recovery, while both Illumina and Nanopore sequencing generated high-quality phage genomes suitable for therapeutic characterisation. Nanopore sequencing provided assemblies comparable to Illumina with minimal benefit from hybrid polishing, supporting its routine use for phage genomics. These findings provide practical guidance for phage genome characterisation workflows and contribute to the development of standardised, regulatory-grade approaches for therapeutic phage assessment.

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West Nile virus in Italy: history and evolving transmission patterns

Giovanetti, M.; Cella, E.; Fonseca, V.; Moir, M.; Oude Munnink, B.; de Martinis, C.; Moreno, A.; Rizzo, A.; Mileto, D.; Caccuri, F.; Caruso, A.; Tramontano, E.; Nanev Slavov, S.; Bispo de Filippis, A. M.; de Oliveira, T.; Alcantara, L. C. J.; Marcello, A.; Colizzi, V.; Rezza, G.; Ciccozzi, M.; Castilletti, C.; Holmes, E. C.; Maggi, F.; Barzon, L.; Lourenco, J.

2026-07-15 infectious diseases 10.64898/2026.07.12.26357540 medRxiv
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West Nile virus (WNV) has become an important public health concern in Europe. Italy is one of the most affected countries, yet our understanding of WNV epidemiology, genomics, and dispersal across hosts and geographic regions is incomplete. AIM: To reveal the history of WNV in Italy by integrating epidemiological, genomic, and environmental data into descriptive and quantitative assessments of its past spatio-temporal surveillance and expansion. We collated vertebrate and mosquito WNV records from national surveillance and the scientific literature spanning multiple decades. Historical serological and molecular data were summarized by host and region, climatic associations with case trends were assessed using regression models, and phylodynamic and phylogeographic analyses reconstructed viral introductions and dispersal within Italy.WNV circulation in Italy has changed markedly over time, with increasing human case reporting and expansion beyond historically affected northern regions. Climate-informed regression models explained recent reporting trends, supporting an environmental contribution to transmission. Phylodynamic analyses identified multiple independent introductions and sustained local transmission with increasing regional connectivity. Wavefront analyses revealed lineage-specific dispersal patterns associated with seasonal climatic gradients. Discrepancies between epidemiological records and genomic sampling highlighted uneven surveillance across regions and host species.WNV emergence in Italy reflects repeated viral introductions, local persistence, heterogeneous surveillance, and environmentally associated dispersal dynamics. Strengthening integrated surveillance combining epidemiological, environmental, and genomic data will improve early detection, the monitoring of transmission dynamics, and public health preparedness under ongoing environmental change.

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Emerging parvovirus associated with an outbreak in Dutch pig farms also detected in pigs and wildlife in Denmark

Canuti, M.; Juncher Hoeg, F.; Vedsted Hammer, A. S.; Kare Jensen, T.; Lauge Quaade, M.; Ryt-Hansen, P.; Droce, A.; Salomonsen, C. M.; Sorensen, S. S.; Larsen, L. E. E.

2026-06-25 microbiology 10.64898/2026.06.24.734170 medRxiv
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A parvovirus recently associated with an outbreak in Dutch pigs was found in Denmark in symptomatic pigs and in fox (Vulpes vulpes) feces and spleens. Pig viruses were more closely related to each other than to viruses found in the respective local wildlife, suggesting a link between the farm outbreaks.

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The HSV-1 immediate early protein ICP22 interacts with the human antisense function 1 protein to promote viral replication

Ye, Y.; Yang, Z.; Xue, M.; Zheng, C.

2026-06-25 microbiology 10.64898/2026.06.24.734377 medRxiv
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Herpes simplex virus type 1 (HSV-1) is a common human pathogen that undergoes lytic replication in epithelial and other permissive cell types and can establish latency in peripheral neurons. ICP22 is a multifunctional HSV-1 immediate-early protein that localizes to the nucleus of infected cells; however, its interactions with host cellular factors remain incompletely understood. Here, ICP22 was demonstrated to interact with the human antisense function 1 protein (ASF1), including both ASF1a and ASF1b, in transfected cells and HSV-1-infected cells, respectively. ICP22 also colocalized with ASF1 in the nucleus. ICP22 amino acids 213 to 340 are important for the interaction of ICP22 with ASF1, whereas amino acids 37 to 153 of ASF1a and ASF1b are critical for their interactions with ICP22. Furthermore, ICP22 expression was associated with reduced ASF1-H3.1 co-immunoprecipitation under the tested conditions. ASF1 knockdown also reduced HSV-1-BAC-Luc luciferase output, indicating that ASF1 contributes to efficient infection-associated reporter activity in this study. Collectively, these results indicate that the interaction of HSV-1 ICP22 with ASF1 might help regulate the transcription of viral or cellular genes during HSV-1 infection. Keywords: HSV-1, ICP22, ASF1, histone H3.

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Lassa fever epidemiology and predictors of mortality in Ebonyi State, Nigeria - A five-year retrospective analysis from 2019 to 2023

Nwojiji, E. C.; Nwambeke, N. O.; Shih, P.-W.; Chukwunenye, C. U.; Odeh, E. C.; Ekuma, M. I.; Azuogu, B.; Iroezindu, M. O.; Liesenborghs, L.; Dijck, C. V.

2026-07-06 epidemiology 10.64898/2026.07.02.26357189 medRxiv
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Lassa fever (LF) is a viral disease that is widespread throughout West Africa, with significant global health consequences. Nigeria bears the highest burden (1309 confirmed cases in 2024) of LF among all endemic countries. Within Nigeria, Ebonyi State bears a high burden of cases. Clinical observations suggest that there may be an increase in the geographic spread and case fatality rate of LF across the State. Ebonyi State case-based data on confirmed cases from 2019 to 2023 collated on the Nigeria Centre for Disease Control and Prevention (NCDC) Surveillance, Outbreak Response Management and Analysis System (SORMAS) platform were analyzed. Descriptive statistics, spatial distribution and time series analysis were performed. Multivariate logistic regression analysis was used to identify predictors of LF mortality and factors associated with PCR positivity. A total of 1,624 suspected cases were reported, 1,343 and 273 were laboratory negative and positive respectively, while 8 probable cases were reported. The yearly number of cases remained stable throughout the study period. Out of the 273 confirmed, 107 died from LF, resulting in a case fatality rate (CFR) of 39.2%. CFR increased non-significantly over time, ranging from 28.6% to 55.8%. Variations in geographic distribution were observed; in 2019 ten local government areas (LGAs) were affected compared to twelve in 2020. A higher incidence was observed between January and March annually. Age above 44 years, bleeding and seizures were significant predictors of mortality. Lower incidence of cases was consistently reported in the Southern LGAs. PCR positivity was associated with individuals who reside in Ebonyi LGA and who have had contact with confirmed cases. The increase in CFR and identification of high-burden areas will help shape policies, allocate resources and provide actionable intervention strategies to combat LF in the State.

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Transmission dynamics of Nipah virus in Bangladesh and India, 2001-2026: systematic review and inference on reproduction number, offspring dispersion, and serial interval

Kim, S.; Mogasale, V. V.; Vesga, J. F.; Kang, H.; Skrip, L.; Jung, S.-m.; Islam, A.; Endo, A.; Edmunds, W. J.; Abbas, K.

2026-07-19 epidemiology 10.64898/2026.07.16.26357631 medRxiv
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Background Nipah virus (NiV) is a priority zoonotic pathogen causing high-fatality outbreaks. Early NiV outbreaks in Malaysia and Singapore had limited transmission beyond spillover events. However, since 2001, NiV outbreaks with person-to-person transmission have occurred in Bangladesh and India, driven by the NiV-Bangladesh genotype and NiV-India genotype. Our study aims to estimate the reproduction number, offspring dispersion, and serial interval governing NiV transmission in Bangladesh and India during 2001-2026. Methods We conducted a systematic review of NiV outbreak investigations in Bangladesh and India, searching PubMed, Embase, Web of Science, and grey literature through 28 February 2026. Case-level offspring counts from 27 eligible sources (323 cases across 67 outbreaks) were used as input to a hierarchical Bayesian negative binomial offspring distribution model. The serial interval was estimated by parametric distribution fitting to 137 transmission pairs. Country-stratified and sensitivity analyses were performed to evaluate the robustness of estimates. Results Pooling across 67 outbreaks, we estimated a median reproduction number of 0.46 (95% CrI: 0.28-0.73), an offspring dispersion parameter of 0.07 (0.05-0.10), and a serial interval of 13.3 days (95% CI: 12.8-13.8). Country-stratified median reproduction numbers were 0.48 (0.23-0.97) for India and 0.35 (0.19-0.59) for Bangladesh, and dispersion parameters were 0.04 (0.02-0.07) and 0.11 (0.06-0.18), respectively, indicating marked overdispersion in both settings. Conclusion NiV transmission is self-limiting on average and highly overdispersed, suggesting that a disproportionate share of onward transmission arises from a small number of cases. This epidemiological profile supports targeted containment measures, including contact tracing and quarantine, for effective NiV outbreak control.

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Anti-Human T-Lymphotropic Virus Type 1 (Htlv-1) Seropositivity In Haematological Malignancies At A Major Clinical Setting In Ghana

Awuku, F.; Omoniyi, P.; Adjei, D. N.; Seshie, M.; Sagoe, K. W. C.; Kuma, A. A. B.-A.

2026-07-10 infectious diseases 10.64898/2026.07.07.26357496 medRxiv
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Background Human T-cell lymphotropic virus - 1 (HTLV-1) is the causative agent of Adult T-cell Leukaemia/Lymphoma (ATLL), a malignancy of CD4+ cells, and HTLV-1-associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP), a demyelinating disease. Globally, 10-20 million people are infected, though most remain asymptomatic and about 5% progress to severe disease. Transmission occurs mainly through breastfeeding, sexual contact, contaminated needles, and blood transfusion. In Ghana, evidence on the role of HTLV-1 in haematological malignancies remains scarce. Methods This was a cross-sectional study involving 200 patients with haematological malignancies (Acute Lymphoblastic Leukaemia - 4, Acute Myeloid Leukaemia - 6, Chronic Lymphocytic Leukaemia - 27, Chronic Myeloid Leukaemia - 63, Hodgkin Lymphoma - 21, Multiple Myeloma - 31, Myelodysplasia - 6, Myeloproliferative Neoplasm - 11) at the Haematology Day Care of the Korle-Bu Teaching Hospital. After informed consent was obtained, sera from study participants were tested for anti-HTLV-1 using MP Diagnostics GmbH ELISA immunoassay. Data were analysed using R software version 4.0.2 and SPSS version 31.0.0. Results The study population had a mean age of 49.1{+/-}17.7 years, with majority being females (n=109, 54.5%). Of the 200 samples, 16 (8.0%) were seropositive for HTLV-1, and these were detected in 4 males and 12 females. No statistically significant association was found between HTLV-1 infection and haematological malignancy (exact p = 0.061), sex (p=0.061), and history of blood transfusion (exact p= 1.000). Conclusion The findings show the seroprevalence of HTLV-1 of 8.0% among patients with haematological malignancies. Although there was no probable association between HTLV-1 and haematological malignancies, screening for HTLV-1 in patients with haematological malignancies may help to unravel the exact contribution in these conditions.