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Botrytis cinerea is one of the most important plant-pathogenic fungus. Products based on microorganisms can be used in biocontrol strategies alternative to chemical control, and mycoviruses have been explored as putative biological agents in such approaches. Here, we have explored the mycovirome of B. cinerea isolates from grapevine of Italy and Spain to increase the knowledge about mycoviral diversity and evolution, and to search for new widely distributed mycoviruses that could be active ingredients in biological products to control this hazardous fungus. A total of 248 B. cinerea field isolates were used for our metatranscriptomic study. Ninety-two mycoviruses were identified: 62 new mycoviral species constituting putative novel viral genera and families. Of these mycoviruses, 57 had a positive-sense single-stranded RNA (ssRNA) genome, 19 contained a double-stranded RNA (dsRNA) genome, 15 had a negative-sense ssRNA genome, and 1 contained a single-stranded DNA (ssDNA) genome. In general, ssRNA mycoviruses were widely distributed in all sampled regions, the ssDNA mycovirus was more frequently found in Spain, and dsRNA mycoviruses were scattered in some pools of both countries. Some of the identified mycoviruses belong to clades that have never been found associated with Botrytis species: Botrytis-infecting narnaviruses; alpha-like, umbra-like, and tymo-like ssRNA+ mycoviruses; trisegmented ssRNA- mycovirus; bisegmented and tetrasegmented dsRNA mycoviruses; and finally, an ssDNA mycovirus. Among the results obtained in this massive mycovirus screening, the discovery of novel bisegmented viruses, phylogenetically related to narnaviruses, is remarkable.IMPORTANCE The results obtained here have expanded our knowledge of mycoviral diversity, horizontal transfers, and putative cross-kingdom events. To date, this study presents the most extensive and wide diversity collection of mycoviruses infecting the necrotrophic fungus B. cinerea The collection included all types of mycoviruses, with dsRNA, ssRNA+, ssRNA-, and ssDNA genomes, most of which were discovered here, and some of which were previously reported as infecting B. cinerea or other plant-pathogenic fungi. Some of these mycoviruses are reported for the first time here associated with B. cinerea, as a trisegmented ssRNA- mycovirus and as an ssDNA mycovirus, but even more remarkablly, we also describe here four novel bisegmented viruses (binarnaviruses) not previously described in nature. The present findings significantly contribute to general knowledge in virology and more particularly in the field of mycovirology. ; J.R.-R. was partially supported by a postdoctoral fellowship from the Severo Ochoa Program for Centers of Excellence in R&D (2017–2021) from the Agencia Estatal de Investigación of Spain (grant SEV-2016-0672 to the Centro de Biotecnología y Genómica de Plantas). I.G.-C. was supported by contract PEJ-2018-TL/BIO-11747 from Comunidad de Madrid, Spain. This study was completely supported financially by VIROPLANT, a project that received funding from the European Union's Horizon 2020 Research and Innovation Program (grant agreement number 773567) ; Peer reviewed

19 Pág. ; Botrytis virus F (BVF) is a positive-sense, single-stranded RNA (+ssRNA) virus within the Gammaflexiviridae family of the plant-pathogenic fungus Botrytis cinerea. In this study, the complete sequence of a BVF strain isolated from B. cinerea collected from grapevine fields in Spain was analyzed. This virus, in this work BVF-V448, has a genome of 6827 nt in length, excluding the poly(A) tail, with two open reading frames encoding an RNA dependent RNA polymerase (RdRP) and a coat protein (CP). The 5′-and 3′-terminal regions of the genome were determined by rapid amplification of cDNA ends (RACE). Furthermore, a yet undetected subgenomic RNA species in BVF-V448 was identified, indicating that the CP is expressed via 3′ coterminal subgenomic RNAs (sgRNAs). We also report the successful construction of the first BVF full-length cDNA clone and synthesized in vitro RNA transcripts using the T7 polymerase, which could efficiently transfect two different strains of B. cinerea, B05.10 and Pi258.9. The levels of growth in culture and virulence on plants of BVF-V448 transfected strains were comparable to BVF-free strains. The infectious clones generated in this work provide a useful tool for the future development of an efficient BVF foreign gene expression vector and a virus-induced gene silencing (VIGS) vector as a biological agent for the control of B. cinerea. ; This research was funded by Viroplant, a project that received funding from the European Union's Horizon 2020 Research and Innovation Programme (Grant Agreement no. 773567) ; Peer reviewed

Plant RNA (ribonucleic acid) viruses are obligate intracellular parasites with single-stranded (ss) or double-stranded RNA genome(s) generally encapsidated but rarely enveloped. For viruses with ssRNA genomes, the polarity of the infectious RNA (positive or negative) and the presence of one or more genomic RNA segments are the features that mostly determine the molecular mechanisms governing the replication process. RNA viruses cannot penetrate plant cell walls unaided and must enter the cellular cytoplasm through mechanically induced wounds or assisted by a biological vector. After desencapsidation, their genome remains in the cytoplasm where it is translated, replicated and encapsidated in a coupled manner. Replication occurs in large viral replication complexes (VRCs), tethered to modified membranes of cellular organelles and composed by the viral RNA templates and by viral and host proteins. Cis-acting elements located in viral RNA templates regulate the recognition by the virus-encoded RNA-dependent RNA polymerase and possibly contribute to VRC assembly and/or activation. ; Work in the authors' laboratories is supported by grants BFU2012-36095 BIO2013-49053-R and Plant KBBE PCIN-2013-056 from the Ministerio de Economía y Competitividad (MINECO, Spain) and from the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 655841. ; Peer Reviewed

Thosea asigna virus (TaV), an insect virus belonging to the Permutatetraviridae family, has a positive-sense single-stranded RNA (ssRNA) genome with two overlapping open reading frames, encoding for the replicase and capsid proteins. The particular TaV replicase includes a structurally unique RNA-dependent RNA polymerase (RdRP) with a sequence permutation in the palm sub-domain, where the active site is anchored. This non-canonical arrangement of the RdRP palm is also found in double-stranded RNA viruses of the Birnaviridae family. Both virus families also share a conserved VPg sequence motif at the polymerase N-terminus which in birnaviruses appears to be used to covalently link a fraction of the replicase molecules to the 5'-end of the genomic segments. Birnavirus VPgs are presumed to be used as primers for replication initiation. Here we have solved the crystal structure of the TaV RdRP, the first non-canonical RdRP of a ssRNA virus, in its apo- form and bound to different substrates. The enzyme arranges as a stable dimer maintained by mutual interactions between the active site cleft of one molecule and the flexible N-terminal tail of the symmetrically related RdRP. The latter, partially mimicking the RNA template backbone, is involved in regulating the polymerization activity. As expected from previous sequence-based bioinformatics predictions, the overall architecture of the TaV enzyme shows important resemblances with birnavirus polymerases. In addition, structural comparisons and biochemical analyses reveal unexpected similarities between the TaV RdRP and those of Flaviviruses. In particular, a long loop protruding from the thumb domain towards the central enzyme cavity appears to act as a platform for de novo initiation of RNA replication. Our findings strongly suggest an unexpected evolutionary relationship between the RdRPs encoded by these distant ssRNA virus groups. ; Work in Barcelona was supported by grant BIO2011-24333 from the Spanish Ministry of Economy and Competitiveness and by the SILVER Large Scale Collaborative Project, grant agreement number 260644, of the European Union 7th Framework. Work in Madrid was supported by grant AGL2011-24758 from the Spanish Ministry of Economy and Competitiveness. X-ray data were collected at the ESRF (Grenoble, France) within a Block Allocation Group (BAG Barcelona) and at the SLS (Villigen, Switzerland). Financial support was provided by the ESRF and SLS. DSF was supported by a pre-doctoral fellowship from fundació "La Caixa" ; Peer Reviewed

WOS: 000358225100082 ; PubMed ID: 26139522 ; We report the discovery and characterization of a double-stranded RNA (dsRNA) mycovirus isolated from the human pathogenic fungus Aspergillus fumigatus, Aspergillus fumigatus tetramycovirus-1 (AfuTmV-1), which reveals several unique features not found previously in positive-strand RNA viruses, including the fact that it represents the first dsRNA (to our knowledge) that is not only infectious as a purified entity but also as a naked dsRNA. The AfuTmV-1 genome consists of four capped dsRNAs, the largest of which encodes an RNA-dependent RNA polymerase (RdRP) containing a unique GDNQ motif normally characteristic of negative-strand RNA viruses. The third largest dsRNA encodes an S-adenosyl methionine-dependent methyltransferase capping enzyme and the smallest dsRNA a P-A-S-rich protein that apparently coats but does not encapsidate the viral genome as visualized by atomic force microscopy. A combination of a capping enzyme with a picorna-like RdRP in the AfuTmV-1 genome is a striking case of chimerism and the first example (to our knowledge) of such a phenomenon. AfuTmV-1 appears to be intermediate between dsRNA and positive-strand ssRNA viruses, as well as between encapsidated and capsidless RNA viruses. ; Thai Government education program; Turkish Government education program; Leverhulme TrustLeverhulme Trust; Erasmus European Union Programme ; We thank Ren Zhang (Commonwealth Scientific and Industrial Research Organisation) and Matt Dickinson (University of Nottingham) for sequence data of flax rust Melampsora lini dsRNAs, and Profs. Eugene Koonin and Valerian Dolja for their advice on phylogenetic analyses. L.K. and S.O. thank the Thai and the Turkish Government education programs, respectively, for supporting their PhD programs. R.H.A.C. thanks The Leverhulme Trust for an Emeritus Fellowship award, and I.K.-L. acknowledges the support of the Erasmus European Union Programme.

To compare the secondary structure profiles of RNA molecules we developed the CROSSalign method. CROSSalign is based on the combination of the Computational Recognition Of Secondary Structure (CROSS) algorithm to predict the RNA secondary structure profile at single-nucleotide resolution and the Dynamic Time Warping (DTW) method to align profiles of different lengths. We applied CROSSalign to investigate the structural conservation of long non-coding RNAs such as XIST and HOTAIR as well as ssRNA viruses including HIV. CROSSalign performs pair-wise comparisons and is able to find homologs between thousands of matches identifying the exact regions of similarity between profiles of different lengths. In a pool of sequences with the same secondary structure CROSSalign accurately recognizes repeat A of XIST and domain D2 of HOTAIR and outperforms other methods based on covariance modeling. The algorithm is freely available at the webpage http://service.tartaglialab.com//new_submission/crossalign. ; The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013), through the European Research Council, under grant agreement RIBOMYLOME_309545 (Gian Gaetano Tartaglia), and from the Spanish Ministry of Economy and Competitiveness (BFU2014-55054-P and BFU2017-86970-P). We also acknowledge support from AGAUR (2014 SGR 00685), the Spanish Ministry of Economy and Competitiveness, Centro de Excelencia Severo Ochoa 2013–2017 (SEV-2012-0208). We also thank the CRG fellowship to SM.

The COVID-19 pandemic has exacerbated existing social inequities (de la Barre et al., 2020; Fenley, 2020; Perri et al., 2020). The closure of drop-in services and community centres, reduced access to public spaces, and a reduction in resources has had a negative impact on marginalized populations (Perri et al., 2020). Moreover, these tumultuous circumstances have led to increased numbers of persons setting up encampments and clustering in public space across British Columbia, Canada (British Columbia Ministry of Health, 2020b). In response to pandemic-related health concerns involving high-density encampments, the Province of British Columbia, in association with various government and not-for-profit stakeholders, activated Emergency Response Centres (ERCs) to house persons experiencing homelessness during COVID-19. At first glance, this innovative, intersectoral response showed promise as a short-term solution to housing our most vulnerable during public health crises. However, forcing people out of public park space has intensified social inequities and reinforced existing power relationships (Fenley, 2020). This case study critically examines the lessons learned during COVID-19 to inform how we advance change towards socially sustainable public spaces. Through the lens of equity, access to public space for vulnerable populations during COVID-19 in Victoria, British Columbia, is explored. A stakeholder analysis is presented to illuminate the nature of stakeholder engagement within the city of Victoria, followed by a review of the intersectoral response that led to the activation of ERCs and the mobilization of hotel rooms to accommodate people experiencing homelessness during the pandemic. Furthermore, this case study discusses how participatory processes, such as equity-centred design, placemaking, and equity mapping, can facilitate community and citizen engagement. This case highlights the emergence of leisure-related innovations as catalysts for social change-an increasingly important area of leisure research. In addition, this case study outlines the urgent need for research related to the intersection of COVID-19, equity, public space, and leisure. For broader audiences, such as local governments, not-for-profit organizations, and leisure service providers, the value of this case study is underscored by the relevance of co-creation in the context of inclusive land-use planning, policy, and design.

"One Health has now expanded beyond discussion of emerging infectious diseases and zoonoses. Retaining its interdisciplinary focus, this new edition includes more non-infectious disease issues and climate change discussion alongside revised case studies and expanded methodology chapters to draw out implications for practice"--

This book aims to provide a series of informative descriptions of key elements of health emergency preparedness and response that are underpinned by real-life examples. The book is divided into 17 chapters, as follows: introduction on the need to prepare for health emergencies; the planning process; risk assessment; writing an emergency plan; emergency planning and response, and working in partnership; interprofessional working and understanding some emotional barriers and unconscious processes that might influence practice in group and team work; command, control and communication; communications during a health emergency; psychosocial and mental health care before, during and after emergencies, disasters and major incidents; business continuity; training and exercising for emergency preparedness, resilience and response; post-incident follow-up; mass casualty incidents; preparedness and response to pandemics and other infectious disease emergencies; chemical, biological, radiological and nuclear incidents; a military case study; and, health and emergencies in Canterbury, New Zealand, from pandemics to earthquakes.

This book, inclusive of 11 chapters, discusses various non-pesticide-based control strategies against several medically-important disease vector species, including those relevant to the control of malaria, dengue, schistosomiasis, and lymphatic filariasis. Each chapter is dedicated to the discussion of a specific strategy, which includes biological control with arthropods, fungi, and larvivorous fishes, control using various plants, control using semiochemicals, environmental control (i.e., house screening, sanitation, water management), and integrated vector management. Successful applications of these various control strategies in the field are described, as well as recommendations for further improvements. This book will be of valuable use to those engaged in tropical medicine and medical entomology.