The Evolution of GII.4 Norovirus and the Sources and Drivers of Norovirus Pandemics

The norovirus genotype GII.4 is a leading cause of human gastroenteritis worldwide and has caused six pandemics since the mid-1990s. In this thesis, we use phylogenetic analyses to investigate the evolutionary history of the GII.4 genotype and the sources and drivers of norovirus pandemics. We first examine the early history of GII.4 and suggest that the increased prevalence of GII.4 concomitant with the first pandemic was a `perfect storm' where a virus capable of accommodating a high level of amino acid change with a high mutation rate enabling efficient transmission acquired a highly stable viral capsid and/or an increased susceptible population size by expanding its receptor-binding repertoire. We next reconstruct the temporal history of GII.4 and demonstrate that each pandemic strain circulated undetected within poorly sampled reservoir populations for years prior to pandemic emergence. Over several years prior to pandemic emergence, the strain diversifies into a large number of lineages and spatiotemporal reconstruction suggests the strain undergoes low-level worldwide circulation. This indicates that the viral genetic changes important for pandemic emergence are acquired years prior to the pandemic and are therefore not the proximal driver of pandemic spread; we hypothesise that genetic changes pre-adapt the strain for future emergence by shifting the virus to a new region of antigenic space. We demonstrate significant amino acid diversity within pandemic strains, with highly diverse sites within a strain often coinciding with immune epitopes and/or receptor-binding regions. This diversity begins to be accumulated prior to pandemic emergence. We hypothesise that increasing host population immunity curtails circulation of the preceding pandemic strain and results in a new pandemic by opening a niche into which many closely related but subtly different viral lineages can emerge. Finally, we examine two newly emerging norovirus strains and demonstrate that they share polymerase substitutions that may enable increased transmission

Cover crop productivity and subsequent soybean yield in the western Corn Belt

Cover crops (CC) in corn (Zea mays L.) and soybean [Glycine max (L.) Merr.] rotations may prevent N loss and provide other ecosystem services but CC productivity in the western Corn Belt is limited by the short growing season. Our objective was to assess CC treatment and planting practice effects on CC biomass, spring soil nitrate concentrations, and soybean yield at two rainfed sites in eastern and one irrigated site in south-central Nebraska over 4 yr. Cover crop treatments (cereal rye [Secale cereale L.] [RYE] and a mix of rye, legume, and brassica species [MIX]) were planted by broadcast interseeding into corn stands in September (pre-harvest broadcast) or drilling after corn harvest (post-harvest drilled) and terminated 2 wk before planting soybean. Cover crop biomass and N uptake varied between years, but generally at the eastern sites, pre-harvest broadcasting produced more biomass than post-harvest drilling (1.64 and 0.79 Mg ha−1, respectively) and had greater N uptake (37 and 24 kg ha−1, respectively). At the south-central site, post-harvest drilling produced more than pre-harvest broadcasting (1.44 and 1.20 Mg ha−1, respectively). RYE had more biomass than MIX (1.41 and 1.09 Mg ha−1, respectively), but the same N uptake. Soil nitrate reductions after CC were small. In 3 of 12 site-years, soybean yielded less after pre-harvest CC. Yield reductions were not correlated to CC biomass, but were likely due to greater weed pressure. High CC productivity is necessary for high N uptake, and requires site-specific selection of planting practice and CC treatments

Cover crop planting practices determine their performance in the U.S. Corn Belt

Cover crop growing periods in the western U.S. Corn Belt could be extended by planting earlier. We evaluated both pre-harvest broadcast interseeding and post-harvest drilling of the following cover crops: (a) cereal rye (Secale cereale L.) [RYE]; (b) a mix of rye + legumes + brassicas [MIX1], (c) a mix of rye + oat [Avena sativa L.] + legumes + brassicas (MIX2), (d) legumes [LEGU]) and (e) a no cover crop control. These were tested in continuous corn (Zea mays L.) [corn–corn] and soybean [Glycine max (L.) Merr.]–corn systems [soybean–corn] at three sites in Nebraska for their effect on cover crop productivity, soil nutrients, and subsequent corn performance. At the sites with wet fall weather, pre-harvest broadcasting increased cover crop biomass by 90%, to 1.29 Mg ha−1 for RYE and 0.87 Mg ha−1 for MIX1 in soybean–corn, and to 0.56 Mg ha−1 and 0.39 Mg ha−1 in corn–corn, respectively. At the drier site, post-harvest drilling increased biomass of RYE and MIX1 by 95% to 0.80 Mg ha−1 in soybean–corn. Biomass N uptake was highest in pre-harvest RYE and MIX1 at two sites in soybean–corn (35 kg ha−1). RYE and sometimes mixes reduced soil N, but effects on P, K, and soil organic C were inconsistent. In soybean–corn, corn yields decreased by 4% after RYE, and in corn–corn, by 4% after pre-harvest cover crops. Site-specific selection of cover crops and planting practices can increase their performance while minimizing impacts on corn

Producing polished prokaryotic pangenomes with the Panaroo pipeline

Population-level comparisons of prokaryotic genomes must take into account the substantial differences in gene content resulting from horizontal gene transfer, gene duplication and gene loss. However, the automated annotation of prokaryotic genomes is imperfect, and errors due to fragmented assemblies, contamination, diverse gene families and mis-assemblies accumulate over the population, leading to profound consequences when analysing the set of all genes found in a species. Here, we introduce Panaroo, a graph-based pangenome clustering tool that is able to account for many of the sources of error introduced during the annotation of prokaryotic genome assemblies. Panaroo is available at https://github.com/gtonkinhill/panaroo.Peer reviewe

Dissemination of Mycobacterium abscessus via global transmission networks.

Mycobacterium abscessus, a multidrug-resistant nontuberculous mycobacterium, has emerged as a major pathogen affecting people with cystic fibrosis (CF). Although originally thought to be acquired independently from the environment, most individuals are infected with one of several dominant circulating clones (DCCs), indicating the presence of global transmission networks of M. abscessus. How and when these clones emerged and spread globally is unclear. Here, we use evolutionary analyses of isolates from individuals both with and without CF to reconstruct the population history, spatiotemporal spread and recent transmission networks of the DCCs. We demonstrate synchronous expansion of six unrelated DCCs in the 1960s, a period associated with major changes in CF care and survival. Each of these clones has spread globally as a result of rare intercontinental transmission events. We show that the DCCs, but not environmentally acquired isolates, exhibit a specific smoking-associated mutational signature and that current transmission networks include individuals both with and without CF. We therefore propose that the DCCs initially emerged in non-CF populations but were then amplified and spread through the CF community. While individuals with CF are probably the most permissive host, non-CF individuals continue to play a key role in transmission networks and may facilitate long-distance transmission.Funding for this work was provided by The Wellcome Trust (investigator award no. 107032/Z/15/Z to R.A.F.), Fondation Botnar (Programme grant no. 6063) and the UK CF Trust (Innovation Hub award no. 001; Strategic Research Centre award no. 010). M.S., N.A.H. and R.M.D. acknowledge the Cystic Fibrosis Foundation for funding

Viral Networks: Connecting Digital Humanities and Medical History

This volume of original essays explores the power of network thinking and analysis for humanities research. Contributing authors are all scholars whose research focuses on a medical history topic—from the Black Death in fourteenth-century Provence to psychiatric hospitals in twentieth-century Alabama. The chapters take readers through a variety of situations in which scholars must determine if network analysis is right for their research; and, if the answer is yes, what the possibilities are for implementation. Along the way, readers will find practical tips on identifying an appropriate network to analyze, finding the best way to apply network analysis, and choosing the right tools for data visualization. All the chapters in this volume grew out of the 2018 Viral Networks workshop, hosted by the History of Medicine Division of the National Library of Medicine (NIH), funded by the Office of Digital Humanities of the National Endowment for the Humanities, and organized by Virginia Tech

A lung-specific mutational signature enables inference of viral and bacterial respiratory niche

Exposure to different mutagens leaves distinct mutational patterns that can allow inference of pathogen replication niches. We therefore investigated whether SARS-CoV-2 mutational spectra might show lineage-specific differences, dependent on the dominant site(s) of replication and onwards transmission, and could therefore rapidly infer virulence of emergent variants of concern (VOCs). Through mutational spectrum analysis, we found a significant reduction in G>T mutations in the Omicron variant, which replicates in the upper respiratory tract (URT), compared to other lineages, which replicate in both the URT and lower respiratory tract (LRT). Mutational analysis of other viruses and bacteria indicates a robust, generalizable association of high G>T mutations with replication within the LRT. Monitoring G>T mutation rates over time, we found early separation of Omicron from Beta, Gamma and Delta, while mutational patterns in Alpha varied consistent with changes in transmission source as social restrictions were lifted. Mutational spectra may be a powerful tool to infer niches of established and emergent pathogens.Fil: Ruis, Christopher. University of Cambridge; Estados UnidosFil: Peacock, Thomas P.. Imperial College London; Reino UnidoFil: Polo Ilacqua, Luis Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Masone, Diego Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Alvarez, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Hinrichs, Angie S.. University Of California At Santa Cruz.; Estados UnidosFil: Turakhia, Yatish. University of California at San Diego; Estados UnidosFil: Cheng, Ye. University of California at San Diego; Estados UnidosFil: McBroome, Jakob. University Of California At Santa Cruz.; Estados UnidosFil: Corbett Detig, Russell. University Of California At Santa Cruz.; Estados UnidosFil: Parkhill, Julian. University of Cambridge; Reino UnidoFil: Floto, R. Andres. University of Cambridge; Reino Unid

Defective ALC1 nucleosome remodeling confers PARPi sensitization and synthetic lethality with HRD.

Chromatin is a barrier to efficient DNA repair, as it hinders access and processing of certain DNA lesions. ALC1/CHD1L is a nucleosome-remodeling enzyme that responds to DNA damage, but its precise function in DNA repair remains unknown. Here we report that loss of ALC1 confers sensitivity to PARP inhibitors, methyl-methanesulfonate, and uracil misincorporation, which reflects the need to remodel nucleosomes following base excision by DNA glycosylases but prior to handover to APEX1. Using CRISPR screens, we establish that ALC1 loss is synthetic lethal with homologous recombination deficiency (HRD), which we attribute to chromosome instability caused by unrepaired DNA gaps at replication forks. In the absence of ALC1 or APEX1, incomplete processing of BER intermediates results in post-replicative DNA gaps and a critical dependence on HR for repair. Hence, targeting ALC1 alone or as a PARP inhibitor sensitizer could be employed to augment existing therapeutic strategies for HRD cancers.Work in I.A.’s group is funded by the WellcomeTrust (grant number 210634), BBSRC (BB/R007195/1), and Cancer ResearchUK (C35050/A22284). Work in D.A.’s group is funded by the Cancer ResearchUK Career Development Fellowship (grant number 16304). Work in the S.J.B.lab is supported by the Coun, which receives its core fundingfrom Cancer Research UK (FC0010048), the UK Medical Research Council(FC0010048), and the Wellcome Trust (FC0010048); a European Research Council (ERC) Advanced Investigator Grant (TelMetab); and Wellcome TrustSenior Investigator and Collaborative Grants. S.S.-B. was the recipient of an EMBO Long Term Fellowship (ALTF 707-2019) and a MSCA individual fellow-ship (grant 886577). Work in the J.R.C. group is funded by CRUK Career Devel-opment Fellowship (C52690/A19270) with infrastructural support from Well-come core award 090532/Z/09/ZS

Stepwise pathogenic evolution of Mycobacterium abscessus.

Although almost all mycobacterial species are saprophytic environmental organisms, a few, such as Mycobacterium tuberculosis, have evolved to cause transmissible human infection. By analyzing the recent emergence and spread of the environmental organism M. abscessus through the global cystic fibrosis population, we have defined key, generalizable steps involved in the pathogenic evolution of mycobacteria. We show that epigenetic modifiers, acquired through horizontal gene transfer, cause saltational increases in the pathogenic potential of specific environmental clones. Allopatric parallel evolution during chronic lung infection then promotes rapid increases in virulence through mutations in a discrete gene network; these mutations enhance growth within macrophages but impair fomite survival. As a consequence, we observe constrained pathogenic evolution while person-to-person transmission remains indirect, but postulate accelerated pathogenic adaptation once direct transmission is possible, as observed for M. tuberculosis Our findings indicate how key interventions, such as early treatment and cross-infection control, might restrict the spread of existing mycobacterial pathogens and prevent new, emergent ones