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Seifert, S. N.

Publications and source records attributed to Seifert, S. N..

17 recordsLinked to original sources

Molecular and ecological determinants of effective reassortment in orthohantaviruses

A central unsolved problem in RNA virus evolution is understanding why some viral reassortants establish and persist while others do not. To answer this question, we reconstructed reassortant histories across 553 viral genomes from seven orthohantavirus species between 1983 and 2024 using phylogenetic reconciliation and molecular dating. We found that the frequency of retained reassortants varied among orthohantaviruses. For example, reassortment ranged from absent in Andes virus to frequent in Dobrava-Belgrade, Sin Nombre, Seoul, Puumala, and Tula viruses, showing that effective reassortment is not a genus-wide constant. Our Bayesian hierarchical models identified local host overlap as the strongest ecological factor associated with viral reassortment, while cross-segment linkage and terminal RNA structure serve as a molecular filter. We found that the probability of reassortment establishment is highest when ecological opportunity is paired with molecular permissiveness, with their interaction term inferred as the strongest signal in our establishment models (posterior probability = 0.97). These results suggest that reassortment in orthohantaviruses is a sequentially filtered evolutionary process in which divergent lineages must first meet in a host through ecological overlap, exchange segments that are molecularly compatible, and do so within a lineage background permissive to establishment in the host population.

evolutionary biology↗

Viral reservoir status in small mammals emerges as a predictable life-history trait after correcting for surveillance bias

Small mammals, particularly rodents and shrews, act as primary reservoirs for Arenaviruses and Hantaviruses, zoonotic pathogens causing substantial global morbidity. However, our understanding of reservoir ecology is obscured by biased surveillance efforts, where sampling preferentially targets synanthropic species and high-income regions. It remains unclear whether observed patterns of reservoir competence, such as the association with synanthropy, are biological realities or artefacts of surveillance bias. We conducted a systematic review and data synthesis of global surveillance efforts (1960-2023), creating a harmonised database of over 590,000 recorded small mammals contributing 716,000 assay results. We then integrated this with macroecological trait data and phylogenetics to model reservoir probability using Bayesian phylogenetic dyadic generalised linear mixed models. We identified substantial taxonomic and geographic biases; surveillance is heavily skewed towards the Palearctic and widespread, large-bodied species, while 46% of host genera remain entirely unsampled. Geographically, surveillance intensity correlates strongly with accessibility and night-light intensity rather than host biodiversity. After statistically correcting for historical sampling volume, we demonstrate that reservoir status is a predictable biological trait. A fast pace of life (e.g., early maturity, large litters) is associated with an increased probability of reservoir status, independent of sampling effort. Synanthropy also remains a strong, independent predictor, indicating that commensal species act as genuine biological amplifiers in modified landscapes. Evolutionary analyses reveal a mosaic of broad lineage-level co-divergence punctuated by frequent, reactive host-switching. By projecting these models globally, we demonstrate that anthropogenic disturbance acts as an ecological filter, fundamentally challenging the assumption that pristine tropical ecosystems represent the highest intrinsic hazard for viral emergence.

ecology↗

Machine Learning Enables Viral Genome-Agnostic Classification of RNA Virus Infections from Host Transcriptomes

Targeted PCR diagnosis of RNA viruses is sequence dependent, meaning that the accuracy of the assay depends on the identity of the viral sequence. However, sequence-targeted assays can miss novel or divergent viruses. We test whether the host transcriptome alone can classify RNA virus infections without using viral sequences. Using publicly available data on Huh7 and Calu-3 cells experimentally infected with diverse negative-sense RNA, we evaluate two host-derived feature sets (differential expression dataset of identified genes and alignment-free nucleotide k-mer spectra) and compare hierarchical clustering with Random Forests. Across datasets and timepoints (12 to 24 hpi), Random forests accurately distinguished cells infected with different viruses. Controls with label permutation and read shuffling across experimental conditions established non-random performance. Influenza A infections exhibited the strongest, most distinct signatures, whereas Ebola and Lassa virus responses were subtler yet still classifiable. These results show that host-only transcriptomics encodes virus-specific, complex signals that machine learning can exploit, enabling genome-agnostic classification of infection statuses. This approach could aid early outbreak triage when primer based detection methods fail or viral genomes are unknown and complements sequence-based discovery.

microbiology↗

Aminopeptidase N is a receptor for hedgehog merbecoviruses

Merbecoviruses, closely related to the highly pathogenetic Middle East Respiratory Syndrome Coronavirus (MERS-CoV), circulate in hedgehogs throughout Europe and Asia, raising concerns about zoonotic transmission to humans and domestic animals. Unfortunately, how these viruses enter host cells remains unknown, hindering experimental studies. Here, we tested known coronavirus receptor orthologues from European hedgehogs (Erinaceus europaeus) and identified Aminopeptidase N (APN) as an entry receptor for hedgehog merbecoviruses. We confirm this result with single-cycle pseudotype and replication-competent virus experiments as well as protein binding assays. A screen of 30 mammalian APN orthologues reveals restricted cross-species receptor use. Cryo-electron microscopy analysis of the viral glycoprotein-receptor complex shows a unique interface distinct from known coronavirus spike:APN interactions, providing a molecular basis for species barriers. These findings expand the known range of receptor use not only within merbecoviruses but also betacoronaviruses, improving our understanding of betacoronavirus receptors, and informing risk assessments for viral emergence.

microbiology↗

TATAT: a containerized software for generating annotated coding transcriptomes from raw RNA-seq data

MotivationMany transcriptome creation workflows are not standardized, are difficult to install or share, prone to breaking as dependencies update or cease to be maintained and are resource intensive. Due to a lack of authoritative literature, many also overlook potentially important steps, such as thinning contig over-assembly or identifying transcript consensus across samples, which reduce resource demands during annotation and increase the accuracy of final transcripts. ResultsWe developed TATAT, a modular, Dockerized software that contains all the tools necessary to generate an annotated coding transcriptome from raw RNA-seq data. The tools remain in a static state and can be coordinated with bash and python scripts provided therein, making TATAT a standardized, reproducible workflow that can easily be shared and installed. We preferentially incorporate tools that are not only accurate, but are fast and require less RAM, and subsequently show TATAT can generate a comprehensive transcriptome for a non-model organism, the Egyptian rousette bat (Rousettus aegyptiacus), in [~]8 hours in a high-performance computing (HPC) environment. Availability and implementationThe TATAT code, instructions, and tutorial are available at https://github.com/viralemergence/tatat.

bioinformatics↗

229E- and NL63-like coronaviruses in phyllostomid bats, Belize

We report novel and previously identified alphacoronavirus (-CoV) diversity in three phyllostomid bat species (Desmodus rotundus, Carollia sowelli, and Sturnira parvidens) in Belize. Phylogenetic analysis suggests strong similarity between two Neotropical bat -CoV lineages and human CoVs (229E and NL63) compared to prior bat -CoVs.

ecology↗

Comparative susceptibility of Old World and New World bat cell lines to Zika virus: Insights into viral replication and inflammatory responses

BackgroundFrom the first isolation of Zika virus (ZIKV) in Uganda in 1947, ZIKV had primarily been associated with sporadic human cases in Africa and Asia until ZIKV emerged as an epidemic in the Americas in 2015. As ZIKV spread into new geographic regions, it now has the potential to interact with many novel potential host species whose susceptibility to the virus has yet to be determined. Bats, with their ability to fly and live in or near human structures, are plausible ZIKV reservoirs. However, their competence as hosts for ZIKV remains unresolved. In this study, we investigate the immune response of Old World and New World bats to ZIKV infection in vitro. ResultsWe demonstrate that Egyptian Rousette bat (Rousettus aegyptiacus; Old World) cells are susceptible to ZIKV, while we observed little to no ZIKV replication in Jamaican fruit bat (Artibeus jamaicensis) cells. Notably, both the Asian and African ZIKV lineages elicited a strong proinflammatory response in the R. aegyptiacus cell line including upregulation of IL6, CXCL8, and CCL5. These data contrast with the dampened inflammatory response detected in these bats to other viruses. ConclusionsThe findings reveal that R06E cells derived from Egyptian Rousette bats exhibit robust proinflammatory and antiviral responses upon Zika virus (ZIKV) infection, characterized by significant upregulation of proinflammatory cytokines. This suggests that while these cells support productive ZIKV replication, they also mount a strong immune response, challenging the notion of these bats as immune-tolerant reservoirs and indicating a more complex interaction with the virus.

microbiology↗

Protocol for the production of an Arenavirus and Hantavirus host-pathogen database: Project ArHa.

1Arenaviruses and Hantaviruses, primarily hosted by rodents and shrews, represent significant public health threats due to their potential for zoonotic spillover into human populations. Despite their global distribution, the full impact of these viruses on human health remains poorly understood, particularly in regions like Africa, where data is sparse. Both virus families continue to emerge, with pathogen evolution and spillover driven by anthropogenic factors such as land use change, climate change, and biodiversity loss. Recent research highlights the complex interactions between ecological dynamics, host species, and environmental factors in shaping the risk of pathogen transmission and spillover. This underscores the need for integrated ecological and genomic approaches to better understand these zoonotic diseases. A comprehensive, spatially and temporally explicit dataset, incorporating host-pathogen dynamics and human disease data, is crucial for improving risk assessments, enhancing disease surveillance, and guiding public health interventions. Such a dataset (ArHa) would also support predictive modelling efforts aimed at mitigating future spillover events. This paper proposes the development of this unified database for small-mammal hosts of Arenaviruses and Hantaviruses, identifying gaps in current research and promoting a more comprehensive understanding of pathogen prevalence, spillover risk, and viral evolution. 2 Strengths and Limitations of this studyO_LIThis dataset combines detailed spatial and temporal information, providing a unique resource for understanding geographic and temporal trends in Arenavirus and Hantavirus host-pathogen relationships. C_LIO_LIBy explicitly quantifying sampling biases and detection efforts, the dataset allows more robust and accurate asssessments of pathogen prevalence and distribution. C_LIO_LIThe dataset offers a platform for linking ecological data with human health outcomes, supporting the identification of spillover hotspots. C_LIO_LIThe dataset relies on published material, which may vary in terms of detail, accuracy and completeness. Missing or imprecise information may limit the reliability of subsequent analyses. C_LIO_LIThe dataset will be produced as a static resource which could limit its relevance over time as emerging data will not be added. C_LI

microbiology↗

Viral susceptibility and innate immune competency of Carollia perspicillata bat cells produced for virological studies

Multiple viruses that are highly pathogenic in humans are known to have evolved in bats. How bats tolerate infection with these viruses, however, is poorly understood. As viruses engage in a wide range of interactions with their hosts, it is essential to study bat viruses in a system that resembles their natural environment like bat-derived in vitro cellular models. However, stable and accessible bat cell lines are not widely available for the broader scientific community. Here, we generated in vitro reagents for the Sebas short-tailed bat (Carollia perspicillata), tested multiple methods of immortalization, and characterized their susceptibility to virus infection and response to immune stimulation. Using a pseudotyped virus library and authentic virus infections, we show that these C. perspicillata cell lines derived from a diverse array of tissues are susceptible to viruses bearing the glycoprotein of numerous orthohantaviruses, including Andes and Hantaan virus and are also susceptible to live hantavirus infection. Furthermore, stimulation with synthetic double-stranded RNA prior to infection with VSV and MERS-CoV induced a protective antiviral response, demonstrating the suitability of our cell lines to study the bat antiviral immune response. Taken together, the approaches outlined here will inform future efforts to develop in vitro tools for virology from non-model organisms and these C. perspicillata cell lines will enable studies on virus-host interactions in bats.

microbiology↗

Using serosurveys to optimize surveillance for zoonotic pathogens

Zoonotic pathogens pose a significant risk to human health, with spillover into human populations contributing to chronic disease, sporadic epidemics, and occasional pandemics. Despite the widely recognized burden of zoonotic spillover, our ability to identify which animal populations serve as primary reservoirs for these pathogens remains incomplete. This challenge is compounded when prevalence reaches detectable levels only at specific times of year. In these cases, statistical models designed to predict the timing of peak prevalence could guide field sampling for active infections. Thus, we develop a general model that leverages routinely collected serosurveillance data to optimize sampling for elusive pathogens by predicting time windows of peak prevalence. Using simulated data sets, we show that our methodology reliably identifies times when pathogen prevalence is expected to peak. Then, we demonstrate an implementation of our method using publicly available data from two putative Ebolavirus reservoirs, straw-colored fruit bats (Eidolon helvum) and hammer-headed bats (Hypsignathus monstrosus). We envision our method being used to guide the planning of field sampling to maximize the probability of detecting active infections, and in cases when longitudinal data is available, our method can also yield predictions for the times of year that are most likely to produce future spillover events. The generality and simplicity of our methodology make it broadly applicable to a wide range of putative reservoir species where seasonal patterns of birth lead to predictable, but potentially short-lived, pulses of pathogen prevalence. AUTHOR SUMMARYMany deadly pathogens, such as Ebola, Rabies, Lassa, and Nipah viruses, originate in wildlife and jump to human populations. When this occurs, human health is at risk. At the extreme, this can lead to pandemics such as the West African Ebola epidemic and the COVID-19 pandemic. Despite the widely recognized risk wildlife pathogens pose to humans, identifying host species that serve as primary reservoirs for many pathogens remains challenging. A key obstacle to confirming reservoir hosts is sampling animals with active infections. Often, disease prevalence fluctuates seasonally in wildlife populations and only reaches detectable levels at certain times of year. In these cases, statistical models designed to predict the timing of peak prevalence could guide efficient field sampling for active infections. Therefore, we have developed a general model that uses serological data to predict times of year when pathogen prevalence is likely to peak. We demonstrate with simulated data that our method produces reliable predictions, and then demonstrate an application of our method on two hypothesized reservoirs for Ebola virus, straw-colored fruit bats and hammer-headed bats. Our method can be broadly applied to a range of potential reservoir species where seasonal patterns of birth can lead to predictable pulses of peak pathogen prevalence. Overall, our method can guide future sampling of reservoir populations and can also be used to make predictions for times of year for which future outbreaks in human populations are most likely to occur.

ecology↗

Viral genomic features predict orthopoxvirus reservoir hosts

Orthopoxviruses (OPVs), including the causative agents of smallpox and mpox have led to devastating outbreaks in human populations worldwide. However, the discontinuation of smallpox vaccination, which also provides cross-protection against related OPVs, has diminished global immunity to OPVs more broadly. We apply machine learning models incorporating both host ecological and viral genomic features to predict likely reservoirs of OPVs. We demonstrate that incorporating viral genomic features in addition to host ecological traits enhanced the accuracy of potential OPV host predictions, highlighting the importance of host-virus molecular interactions in predicting potential host species. We identify hotspots for geographic regions rich with potential OPV hosts in parts of southeast Asia, equatorial Africa, and the Amazon, revealing high overlap between regions predicted to have a high number of potential OPV host species and those with the lowest smallpox vaccination coverage, indicating a heightened risk for the emergence or establishment of zoonotic OPVs. Our findings can be used to target wildlife surveillance, particularly related to concerns about mpox establishment beyond its historical range.

microbiology↗

Severe acute respiratory disease in American mink (Neovison vison) experimentally infected with SARS-CoV-2

An animal model that fully recapitulates severe COVID-19 presentation in humans has been a top priority since the discovery of SARS-CoV-2 in 2019. Although multiple animal models are available for mild to moderate clinical disease, a non-transgenic model that develops severe acute respiratory disease has not been described. Mink experimentally infected with SARS-CoV-2 developed severe acute respiratory disease, as evident by clinical respiratory disease, radiological, and histological changes. Virus was detected in nasal, oral, rectal, and fur swabs. Deep sequencing of SARS-CoV-2 from oral swabs and lung tissue samples showed repeated enrichment for a mutation in the gene encoding for nonstructural protein 6 in open reading frame 1a/1ab. Together, these data indicate that American mink develop clinical features characteristic of severe COVID19 and as such, are uniquely suited to test viral countermeasures. One Sentence SummarySARS-CoV-2 infected mink develop severe respiratory disease that recapitulates some components of severe acute respiratory disease, including ARDS.

microbiology↗

A sarbecovirus found in Russian bats uses human ACE2

Spillover of sarbecoviruses from animals to humans has resulted in outbreaks of severe acute respiratory syndrome SARS-CoVs and the ongoing COVID-19 pandemic. Efforts to identify the origins of SARS-CoV-1 and -2 has resulted in the discovery of numerous animal sarbecoviruses - the majority of which are only distantly related to known human pathogens and do not infect human cells. The receptor binding domain (RBD) on sarbecoviruses engages receptor molecules on the host cell and mediates cell invasion. Here, we tested the receptor tropism and serological cross reactivity for RBDs from two sarbecoviruses found in Russian horseshoe bats. While these two viruses are in a viral lineage distinct from SARS-CoV-1 and -2, one virus, Khosta-2, was capable of using human ACE2 to facilitate cell entry. Viral pseudotypes with a recombinant, SARS-CoV-2 spike encoding for the Khosta 2 RBD were resistant to both SARS-CoV-2 monoclonal antibodies and serum from individuals vaccinated for SARS-CoV-2. Our findings further demonstrate that sarbecoviruses circulating in wildlife outside of Asia also pose a threat to global health and ongoing vaccine campaigns against SARS-CoV-2 ONE SENTENCE SUMMARYEuropean bat coronaviruses that are only distantly related to SARS-CoV-2 but use the same cell entry route, escape the immune response against SARS-CoV-2 vaccines, driving the need for broader vaccines.

microbiology↗

Mammal virus diversity estimates are unstable due to accelerating discovery effort

Host-virus association data form the backbone of research into eco-evolutionary drivers of viral diversity and host-level zoonotic risk. However, knowledge of the wildlife virome is inherently constrained by historical discovery effort, and there are concerns that the reliability of ecological inference from host-virus data may be undermined by taxonomic and geographical sampling biases. Here, we evaluate whether current estimates of host-level viral diversity in wild mammals are stable enough to be considered biologically meaningful, by analysing a comprehensive dataset of discovery dates of 6,571 unique mammal host-virus associations between 1930 and 2018. We show that virus discovery rates in mammal hosts are still either constant or accelerating, with little evidence of declines towards viral richness asymptotes in even highly-sampled hosts. Consequently, inference of relative viral richness across host species has been unstable over time, particularly in bats, where intensified surveillance since the early 2000s caused a rapid rearrangement of species ranked viral richness. Our results show that comparative inference of host-level virus diversity across mammals is highly sensitive to even short-term changes in sampling effort. We advise caution to avoid overinterpreting patterns in current data, since our findings suggest that an analysis conducted today could feasibly draw quite different conclusions than one conducted only a decade ago.

ecology↗

Surface-aerosol stability and pathogenicity of diverse MERS-CoV strains from 2012 - 2018

Middle East Respiratory Syndrome coronavirus (MERS-CoV) is a coronavirus that infects both humans and dromedary camels and is responsible for an ongoing outbreak of severe respiratory illness in humans in the Middle East. While some mutations found in camel-derived MERS-CoV strains have been characterized, the majority of natural variation found across MERS-CoV isolates remains unstudied. Here we report on the environmental stability, replication kinetics and pathogenicity of several diverse isolates of MERS-CoV as well as SARS-CoV-2 to serve as a basis of comparison with other stability studies. While most of the MERS-CoV isolates exhibited similar stability and pathogenicity in our experiments, the camel derived isolate, C/KSA/13, exhibited reduced surface stability while another camel isolate, C/BF/15, had reduced pathogenicity in a small animal model. These results suggest that while betacoronaviruses may have similar environmental stability profiles, individual variation can influence this phenotype, underscoring the importance of continual, global viral surveillance.

microbiology↗

Lek-associated movement of a putative Ebolavirus reservoir, the Hammer-headed fruit bat (Hypsignathus monstrosus), in northern Republic of Congo

The biology and ecology of Africas largest fruit bat remains largely understudied and enigmatic despite at least two highly unusual attributes. The acoustic lek mating behavior of the hammer-headed bat (Hypsignathus monstrosus) in the Congo basin was first described in the 1970s. Then in the 2000s, molecular testing implicated this species and other fruit bats as potential reservoir hosts for Ebola virus and it was one of only two fruit bat species epidemiologically linked to the 2008 Luebo, Democratic Republic of Congo, Ebola outbreak. Here we share findings from the first pilot study of hammer-headed bat movement using GPS tracking and accelerometry units and a small preceding radio-tracking trial at an apparent lekking site. The radio-tracking revealed adult males had high rates of nightly visitation to the site compared to females (only one visit) and that two of six females day-roosted [~]100 m west of Libonga, the nearest village that is [~]1.6 km southwest. Four months later, in mid-April 2018, five individual bats, comprised of four males and one female, were tracked from two to 306 days, collecting from 67 to 1022 GPS locations. As measured by mean distance to the site and proportion of nightly GPS locations within 1 km of the site (percent visitation), the males were much more closely associated with the site (mean distance 1.4 km; 51% visitation), than the female (mean 5.5 km; 2.2% visitation). Despite the small sample size, our tracking evidence supports our original characterization of the site as a lek, and the lek itself is much more central to male than female movement. Moreover, our pilot demonstrates the technical feasibility of executing future studies on hammer-headed bats that will help fill problematic knowledge gaps about zoonotic spillover risks and the conservation needs of fruit bats across the continent.

ecology↗