Microbiology & Cell Biology

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    Comparative Aerosol and Surface Stability of SARS-CoV-2 Variants of Concern
    (Centers for Disease Control and Prevention, 2023-05) Bushmaker, Trenton; Kwe Yinda, Claude; Morris, Dylan H.; Holbrook, Myndi G.; Gamble, Amandine; Adney, Danielle; Bushmaker, Cara; van Doremalen, Neeltje; Fischer, Robert J.; Plowright, Raina K.; Lloyd-Smith, James O.; Munster, Vincent J.
    SARS-CoV-2 transmits principally by air; contact and fomite transmission may also occur. Variants of concern are more transmissible than ancestral SARS-CoV-2. We found indications of possible increased aerosol and surface stability for early variants of concern, but not for the Delta and Omicron variants. Stability changes are unlikely to explain increased transmissibility.
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    Severe Acute Respiratory Syndrome Coronavirus 2 Is Detected in the Gastrointestinal Tract of Asymptomatic Endoscopy Patients but Is Unlikely to Pose a Significant Risk to Healthcare Personnel
    (Elsevier, 2022-06) Cherne, Michelle D.; Gentry, Andrew B.; Nemudraia, Anna; Nemudryi, Artem; Hedges, Jodi F.; Walk, Heather; Blackwell, Karlin; Snyder, Deann T.; Jerome, Maria; Madden, Wyatt; Hashimi, Marziah; Sebrell, T. Andrew; King, David B.; Plowright, Raina K.; Jutila, Mark A.; Wiedenheft, Blake; Bimczok, Diane
    Background and aims. Recent evidence suggests that the gut is an additional target for severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) infection. However, whether SARS-CoV-2 spreads via gastrointestinal secretions remains unclear. To determine the prevalence of gastrointestinal SARS-CoV-2 infection in asymptomatic subjects, we analyzed gastrointestinal biopsy and liquid samples from endoscopy patients for the presence of SARS-CoV-2. Methods. We enrolled 100 endoscopic patients without known SARS-CoV-2 infection (cohort A) and 12 patients with a previous COVID-19 diagnosis (cohort B) in a cohort study performed at a regional hospital. Gastrointestinal biopsies and fluids were screened for SARS-CoV-2 by polymerase chain reaction (PCR), immunohistochemistry, and virus isolation assay, and the stability of SARS CoV-2 in gastrointestinal liquids in vitro was analyzed. Results. SARS-CoV-2 ribonucleic acid was detected by PCR in the colonic tissue of 1/100 patients in cohort A. In cohort B, 3 colonic liquid samples tested positive for SARS-CoV-2 by PCR and viral nucleocapsid protein was detected in the epithelium of the respective biopsy samples. However, no infectious virions were recovered from any samples. In vitro exposure of SARS-CoV-2 to colonic liquid led to a 4-log-fold reduction of infectious SARS-CoV-2 within 1 hour (P ≤ .05). Conclusion. Overall, the persistent detection of SARS-CoV-2 in endoscopy samples after resolution of COVID-19 points to the gut as a long term reservoir for SARS-CoV-2. Since no infectious virions were recovered and SARS-CoV-2 was rapidly inactivated in the presence of colon liquids, it is unlikely that performing endoscopic procedures is associated with a significant infection risk due to undiagnosed asymptomatic or persistent gastrointestinal SARS-CoV-2 infections.
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    Ecology, evolution and spillover of coronaviruses from bats
    (Springer Science and Business Media LLC, 2021-11) Ruiz-Aravena, Manuel; McKee, Clifton; Gamble, Amandine; Lunn, Tamika; Morris, Aaron; Snedden, Celine E.; Yinda, Claude Kwe; Port, Julia R.; Buchholz, David W.; Yeo, Yao Yu; Faust, Christina; Jax, Elinor; Dee, Lauren; Jones, Devin N.; Kessler, Maureen K.; Falvo, Caylee A.; Crowley, Daniel; Bharti, Nita; Brook, Cara E.; Aguilar, Hector C.; Peel, Alison J.; Restif, Olivier; Schountz, Tony; Parrish, Colin R.; Gurley, Emily S.; Lloyd-Smith, James O.; Hudson, Peter J.; Munster, Vincent J.; Plowright, Raina K.
    In the past two decades, three coronaviruses with ancestral origins in bats have emerged and caused widespread outbreaks in humans, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Since the first SARS epidemic in 2002–2003, the appreciation of bats as key hosts of zoonotic coronaviruses has advanced rapidly. More than 4,000 coronavirus sequences from 14 bat families have been identified, yet the true diversity of bat coronaviruses is probably much greater. Given that bats are the likely evolutionary source for several human coronaviruses, including strains that cause mild upper respiratory tract disease, their role in historic and future pandemics requires ongoing investigation. We review and integrate information on bat–coronavirus interactions at the molecular, tissue, host and population levels. We identify critical gaps in knowledge of bat coronaviruses, which relate to spillover and pandemic risk, including the pathways to zoonotic spillover, the infection dynamics within bat reservoir hosts, the role of prior adaptation in intermediate hosts for zoonotic transmission and the viral genotypes or traits that predict zoonotic capacity and pandemic potential. Filling these knowledge gaps may help prevent the next pandemic.
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    Optimizing noninvasive sampling of a zoonotic bat virus
    (Wiley, 2021-08) Giles, John R.; Peel, Alison J.; Wells, Konstans; Plowright, Raina K.; McCallum, Hamish; Restif, Olivier
    Outbreaks of infectious viruses resulting from spillover events from bats have brought much attention to bat- borne zoonoses, which has motivated increased ecological and epidemiological studies on bat populations. Field sampling methods often collect pooled samples of bat excreta from plastic sheets placed under-roosts. However, positive bias is introduced because multiple individuals may contribute to pooled samples, making studies of viral dynamics difficult. Here, we explore the general issue of bias in spatial sample pooling using Hendra virus in Australian bats as a case study. We assessed the accuracy of different under- roost sampling designs using generalized additive models and field data from individually captured bats and pooled urine samples. We then used theoretical simulation models of bat density and under- roost sampling to understand the mechanistic drivers of bias. The most commonly used sampling design estimated viral prevalence 3.2 times higher than individual- level data, with positive bias 5–7 times higher than other designs due to spatial autocorrelation among sampling sheets and clustering of bats in roosts. Simulation results indicate using a stratified random design to collect 30–40 pooled urine samples from 80 to 100 sheets, each with an area of 0.75–1 m2, and would allow estimation of true prevalence with minimum sampling bias and false negatives. These results show that widely used under- roost sampling techniques are highly sensitive to viral presence, but lack specificity, providing limited information regarding viral dynamics. Improved estimation of true prevalence can be attained with minor changes to existing designs such as reducing sheet size, increasing sheet number, and spreading sheets out within the roost area. Our findings provide insight into how spatial sample pooling is vulnerable to bias for a wide range of systems in disease ecology, where optimal sampling design is influenced by pathogen prevalence, host population density, and patterns of aggregation.
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    From Protein to Pandemic: The Transdisciplinary Approach Needed to Prevent Spillover and the Next Pandemic
    (MDPI AG, 2021-07) Plowright, Raina K.; Hudson, Peter J.
    Pandemics are a consequence of a series of processes that span scales from viral biology at 10−9 m to global transmission at 106 m. The pathogen passes from one host species to another through a sequence of events that starts with an infected reservoir host and entails interspecific contact, innate immune responses, receptor protein structure within the potential host, and the global spread of the novel pathogen through the naive host population. Each event presents a potential barrier to the onward passage of the virus and should be characterized with an integrated transdisciplinary approach. Epidemic control is based on the prevention of exposure, infection, and disease. However, the ultimate pandemic prevention is prevention of the spillover event itself. Here, we focus on the potential for preventing the spillover of henipaviruses, a group of viruses derived from bats that frequently cross species barriers, incur high human mortality, and are transmitted among humans via stuttering chains. We outline the transdisciplinary approach needed to prevent the spillover process and, therefore, future pandemics.
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    Prioritizing surveillance of Nipah virus in India
    (2019-06) Plowright, Raina K.; Becker, Daniel J.; Crowley, Daniel E.; Washburne, Alex D.; Huang, Tao; Nameer, P. O.; Gurley, Emily S.; Han, Barbara A.
    The 2018 outbreak of Nipah virus in Kerala, India, highlights the need for global surveillance of henipaviruses in bats, which are the reservoir hosts for this and other viruses. Nipah virus, an emerging paramyxovirus in the genus Henipavirus, causes severe disease and stuttering chains of transmission in humans and is considered a potential pandemic threat. In May 2018, an outbreak of Nipah virus began in Kerala, > 1800 km from the sites of previous outbreaks in eastern India in 2001 and 2007. Twenty-three people were infected and 21 people died (16 deaths and 18 cases were laboratory confirmed). Initial surveillance focused on insectivorous bats (Megaderma spasma), whereas follow-up surveys within Kerala found evidence of Nipah virus in fruit bats (Pteropus medius). P. medius is the confirmed host in Bangladesh and is now a confirmed host in India. However, other bat species may also serve as reservoir hosts of henipaviruses. To inform surveillance of Nipah virus in bats, we reviewed and analyzed the published records of Nipah virus surveillance globally. We applied a trait-based machine learning approach to a subset of species that occur in Asia, Australia, and Oceana. In addition to seven species in Kerala that were previously identified as Nipah virus seropositive, we identified at least four bat species that, on the basis of trait similarity with known Nipah virus-seropositive species, have a relatively high likelihood of exposure to Nipah or Nipah-like viruses in India. These machine-learning approaches provide the first step in the sequence of studies required to assess the risk of Nipah virus spillover in India. Nipah virus surveillance not only within Kerala but also elsewhere in India would benefit from a research pipeline that included surveys of known and predicted reservoirs for serological evidence of past infection with Nipah virus (or cross reacting henipaviruses). Serosurveys should then be followed by longitudinal spatial and temporal studies to detect shedding and isolate virus from species with evidence of infection. Ecological studies will then be required to understand the dynamics governing prevalence and shedding in bats and the contacts that could pose a risk to public health.
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    A bioinformatic pipeline for identifying informative SNP panels for parentage assignment from RADseq data
    (2018-06) Andrews, Kimberly R.; Adams, Jennifer R.; Cassirer, E. Frances; Plowright, Raina K.; Gardner, Colby; Dwire, Maggie; Hohenlohe, Paul A.; Waits, Lisette P.
    The development of high-throughput sequencing technologies is dramatically increasing the use of single nucleotide polymorphisms (SNPs) across the field of genetics, but most parentage studies of wild populations still rely on microsatellites. We developed a bioinformatic pipeline for identifying SNP panels that are informative for parentage analysis from restriction site-associated DNA sequencing (RADseq) data. This pipeline includes options for analysis with or without a reference genome, and provides methods to maximize genotyping accuracy and select sets of unlinked loci that have high statistical power. We test this pipeline on small populations of Mexican gray wolf and bighorn sheep, for which parentage analyses are expected to be challenging due to low genetic diversity and the presence of many closely related individuals. We compare the results of parentage analysis across SNP panels generated with or without the use of a reference genome, and between SNPs and microsatellites. For Mexican gray wolf, we conducted parentage analyses for 30 pups from a single cohort where samples were available from 64% of possible mothers and 53% of possible fathers, and the accuracy of parentage assignments could be estimated because true identities of parents were known a priori based on field data. For bighorn sheep, we conducted maternity analyses for 39 lambs from five cohorts where 77% of possible mothers were sampled, but true identities of parents were unknown. Analyses with and without a reference genome produced SNP panels with ≥95% parentage assignment accuracy for Mexican gray wolf, outperforming microsatellites at 78% accuracy. Maternity assignments were completely consistent across all SNP panels for the bighorn sheep, and were 74.4% consistent with assignments from microsatellites. Accuracy and consistency of parentage analysis were not reduced when using as few as 284 SNPs for Mexican gray wolf and 142 SNPs for bighorn sheep, indicating our pipeline can be used to develop SNP genotyping assays for parentage analysis with relatively small numbers of loci.
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    Environmental persistence of influenza H5N1 is driven by temperature and salinity: insights from a Bayesian meta-analysis
    (2018-09) Martin, Gerardo A.; Becker, Daniel J.; Plowright, Raina K.
    Environmental persistence of zoonotic pathogens is a key trait that influences the probability of zoonotic spillover. Pathogen survival outside of the host determines the window available for contact with the new recipient host species and the dose of pathogen available to that host. The longer a pathogen survives in the environment, the more disconnected the reservoir and recipient hosts can be in space and time, and the more likely that an infective dose will be available to recipient hosts. Therefore, environmental persistence is a key parameter for mechanistic models needed to predict pathogen spillover. Avian influenza can be transmitted from wildlife to poultry and people in part due to its ability to persist in the environment. Considerable work has been done to quantify trends in avian influenza persistence across environmental conditions, often published in separate studies with separate datasets. In this paper, we quantify the trends and variability of avian influenza viral persistence across environmental conditions by collating disjoint experimental data on viral particle persistence in water across many studies and a range of environmental conditions. The collated data represent 120 estimates from three different studies of the decay rates of highly pathogenic avian influenza H5N1 (90 estimates from Asia and 30 from Europe) in response to temperature, pH, and salinity. We analyzed these data with a Bayesian model to control for biases with random effects and used experimental replicates and R2 estimates of the publication's regression procedures as statistical weights. We found temperature significantly decreases persistence of H5N1 virus in water, and this effect is stronger than that of salinity alone. Salinity interacts with temperature and probably drives the most contrasting persistence scenarios between cold-saline and warm-saline water bodies, where highest and lowest persistence times could occur respectively. Our work provides needed parameters for models that examine the risk of spillover of avian influenza viruses.
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    Persistent infections support maintenance of a coronavirus in a population of Australian bats (Myotis macropus)
    (2017-07) Jeong, J.; Smith, C. S.; Peel, Alison J.; Plowright, Raina K.; Kerlin, D. H.; McBroom, J.; McCallum, Hamish I.
    Understanding viral transmission dynamics within populations of reservoir hosts can facilitate greater knowledge of the spillover of emerging infectious diseases. While bat-borne viruses are of concern to public health, investigations into their dynamics have been limited by a lack of longitudinal data from individual bats. Here, we examine capture-mark-recapture (CMR) data from a species of Australian bat (Myotis macropus) infected with a putative novel Alphacoronavirus within a Bayesian framework. Then, we developed epidemic models to estimate the effect of persistently infectious individuals (which shed viruses for extensive periods) on the probability of viral maintenance within the study population. We found that the CMR data analysis supported grouping of infectious bats into persistently and transiently infectious bats. Maintenance of coronavirus within the study population was more likely in an epidemic model that included both persistently and transiently infectious bats, compared with the epidemic model with non-grouping of bats. These findings, using rare CMR data from longitudinal samples of individual bats, increase our understanding of transmission dynamics of bat viral infectious diseases.
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    Principles and Patterns of Bat Movements: From Aerodynamics to Ecology
    (2017-09) Voigt, Christian C.; Frick, Winifred F.; Holderied, Marc W.; Holland, Richard; Kerth, Gerald; Mello, Marco A. R.; Plowright, Raina K.; Swartz, Sharon; Yovel, Yossi
    Movement ecology as an integrative discipline has advanced associated fields because it presents not only a conceptual framework for understanding movement principles but also helps formulate predictions about the consequences of movements for animals and their environments. Here, we synthesize recent studies on principles and patterns of bat movements in context of the movement ecology paradigm. The motion capacity of bats is defined by their highly articulated, flexible wings. Power production during flight follows a U-shaped curve in relation to speed in bats yet, in contrast to birds, bats use mostly exogenous nutrients for sustained flight. The navigation capacity of most bats is dominated by the echolocation system, yet other sensory modalities, including an iron-based magnetic sense, may contribute to navigation depending on a bat\'s familiarity with the terrain. Patterns derived from these capacities relate to antagonistic and mutualistic interactions with food items. The navigation capacity of bats may influence their sociality, in particular, the extent of group foraging based on eavesdropping on conspecifics' echolocation calls. We infer that understanding the movement ecology of bats within the framework of the movement ecology paradigm provides new insights into ecological processes mediated by bats, from ecosystem services to diseases.
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