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Annual Conference 2017 Annual Conference 2017 3–6 APRIL, EICC, EDINBURGH, UK POSTER ABSTRACT BOOK @MicrobioSoc #Microbio17 Annual Conference 2017 EICC, Edinburgh 3-6 April Posters displayed Monday to Thursday Virus Workshop: Antivirals and Vaccines Virus Workshop: Clinical Virology Network Virus Workshop: Evolution and Virus Populations Virus Workshop: Gene expression and replication Virus Workshop: Innate Immunity Virus Workshop: Morphogenesis, egress and entry Virus Workshop: Pathogenesis Annual Meeting of Protistology-UK society: Intracellular infection and endosymbiosis within protists Aquatic Microbiology Cell biology of pathogen entry into host cells Circadian Rhythms Critical health challenges in medical mycology Epigenetic and Non Coding RNAs in Eukaryotes Synthetic and Systems Biology Approaches to Microbiology Posters displayed on Monday and Tuesday Geomicrobiology Macromolecular Machines Microbial Mechanisms of Plant pathology Prokaryotic Genetics and Genomics Forum Prokaryotic Microbial Infection Forum Posters displayed on Wednesday and Thursday Anaerobes in infection Environmental and Applied Microbiology Forum Heterogeneity and Polymicrobial Interactions in Biofilms Microbial Cell Surfaces Microbial Genomics: Whole Population to Single Cell Microbial Physiology, Metabolism and Molecular Mechanisms Forum Please note: Abstracts are published as received from the authors and are not subject to editing Annual Conference 2017 EICC, Edinburgh 3-6 April Virus Workshop: Antivirals and Vaccines 1 Investigating membrane viral bending proteins in coronavirus replication. Entedar Alsaadi1, Benjamin W.Neuman1 1University of Reading, Reading, UK, 2University of Reading, Reading, UK The coronavirus envelope spike (S) glycoprotein, a class I viral fusion protein, is responsible for the fusion between the membranes of the virus and the host cell. The fusion peptide has yet to be definitively identified, but bioinformatics analysis suggests that at least part of the fusion peptide is located near the amino terminus of the S2 region of S. Potential conserved membrane-modifying peptides were selected based on amino acid conservation, proximity to the membrane and/or Amphipaseek amphipathic helix prediction software. Peptides were tested for membrane-modifying activity in the presence of giant unilamellar vesicles consisting of DPPC, sphingomyelin and cholesterol. A conserved amphipathic helix motif near the amino terminus of the S2 region beginning with the residues IEDLLF showed the strongest membrane-modifying activity of the peptides tested, and were more active than an amphipathic peptide from Influenza A virus M2 protein that has been previously demonstrated to modify membranes. IEDLLF peptides from Mouse hepatitis virus and Middle Eastern respiratory syndrome virus proteins both changed the apparent size and shape of vesicle membranes in a manner consistent with membrane insertion. In contrast, regions bracketing the transmembrane region of S, the amphipathic fusion motor region and conserved motifs of S2 did not show membrane-modifying activity. Together these results suggest that the IEDLLF region that is conserved across the Coronaviridae inserts into host membranes and functions as the fusion peptide of the coronavirus S protein. Please note: Abstracts are published as received from the authors and are not subject to editing Annual Conference 2017 EICC, Edinburgh 3-6 April Virus Workshop: Antivirals and Vaccines 2 Targetting the RNA Packaging Signal-Mediated Assembly of ssRNA Viruses with Drugs. Amy Barker1, Simon White1, German Leonov3, Shabih Shakeel2, Sarah Butcher2, Reidun Twarock3, Peter Stockley1 1University of Leeds, Leeds, UK, 2University of Helsinki, Helsinki, Finland, 3University of York, York, UK Secondary structures in genomic RNA (packaging signals (PSs)) provide scaffolding for cooperative coat protein (CP) recruitment and virus assembly (Stockley et al. 2013). Using bacteriophage MS2 as a model, RNA-binding ligands to its highest affinity PS (TR) were identified. Mitoxantrone (MTX), a clinically approved anti-cancer agent, was found to bind tightly to TR, consistent with its known specificity for RNAs having a 5′ adenosine bulge in the stem (Zheng et al. 2009). Both TR and genomic RNA-mediated MS2 reassembly are readily inhibited by MTX in vitro in a concentration dependent manner. MTX showed specific inhibition of phage infectivity in vivo, although repeated passages in the presence of drug resulted in recovery of wild-type titre. Sequence analysis of these "escape" mutants suggests that their principle target is the essential maturation protein-RNA contact, rather than PS-CP contacts. We have begun to apply similar approaches to the human Parechovirus 1 system in which we have identified up to 60 copies of a PS sequence. The protein side chains that make up PS binding sites are absolutely conserved throughout the Parecho genus, suggesting that in this case the CP would be a good drug target. Molecular modelling of drugs from a clinically approved database identified a series of compounds that are likely to have affinity for the PS binding site. In vitro these ligands appear to block PS binding (Shakeel et al. 2016).These data suggest that it will be possible to develop specific drugs that target specifics of viral assembly involving RNA-CP contacts. Please note: Abstracts are published as received from the authors and are not subject to editing Annual Conference 2017 EICC, Edinburgh 3-6 April Virus Workshop: Antivirals and Vaccines 3 A Preliminary Report: Cervical Human Papilloma Virus Genotype Distribution in Reproductive Aged Women Attending Primary Health Care in Urban Gambia Haddy Bah Camara1, Kumba Sun Mboob2, Fatou Banja2, Foday Ceesay2, Mathew Anyanwu3, Edward Wright1, Patrick Kimmitt1 1Faculty of Science and Technology, Department of Biomedical Science, University of Westminster, London, UK, 2Department of Medical Microbiology, Edward Francis Small Teaching Hospital, Banjul, Gambia, 3Infectious Disease Clinics, Edward Francis Small Teaching Hospital, Banjul, Gambia Persistent infection with high risk human papilloma virus genotype plays a vital role in the development of most cervical cancers and Cervical Intraepithelial Neoplasia (CIN) among sexually active women worldwide. In the Gambia, cervical cancer is the most frequently diagnosed cancer amongst Gambian women representing about 30% of all registered female cancers. The quadrivalent HPV vaccine, which targets HPV genotypes 16, 18, 6 and 11 was introduced in urban Gambia in 2014. The aim of this study is to determine HPV genotype distribution and to determine the efficacy of the quadrivalent vaccine in this population. Sexually active reproductive women (N= 178; 20 - 49 years old) attending the polyclinic were enrolled. A designed questionnaire was administered; endocervical and high vaginal swabs were collected to determine HPV genotype distribution and co-infection with other genital STI pathogens. HPV was determined using the consensus primers PGMY09/11 targeting the late protein (L1) gene. HPV infection rate was 12.4% and the most prevalent high risk genotype was HPV 52. About 60% of participants infected with HPV were co-infected with Ureaplasma. Bivariate analysis shows that Ureaplasma infection, early sexual debut, Low level of education, and female circumcision were risk factors but not significantly associated with HPV (p > 0.05). This study is the first study carried out in urban Gambia and has demonstrated that the quadrivalent vaccine might not be adequate to protect this population from HPV infection as most high risk genotypes identified in this study are not targeted by the quadrivalent vaccine. Please note: Abstracts are published as received from the authors and are not subject to editing Annual Conference 2017 EICC, Edinburgh 3-6 April Virus Workshop: Antivirals and Vaccines 4 Characterization of a novel hepatitis C virus vaccine candidate based on MVA expressing the nearly full- length HCV genome and lacking C6L vaccinia virus gene María Quirós Marín, Juan García-Arriaza, Carmen Gómez, Carlos Óscar Sorzano, Mariano Esteban Rodríguez National Center for Biotechnology, Madrid, Spain Background: Vaccines based in the Modified Vaccinia virus Ankara (MVA) expressing HCV antigens elicited T- cell immune responses in preclinical and clinical studies. However, MVA still contains several immunomodulatory genes that counteract the host antiviral immune response and deletion of some of them is a promising approach for the improvement of MVA-based vaccines. Methods: To improve the immunogenicity of a previously described HCV vaccine candidate (MVA-HCV), based on MVA expressing the nearly full-length HCV genome, the C6L MVA gene, which encodes for an inhibitor of IFN-β was deleted. The resulting vaccine candidate (MVA-HCV ΔC6L) was tested in vitro and in vivo and compared to its parental virus. Results: MVA-HCV ΔC6L expresses all HCV antigens and deletion of C6L had no effect on viral growth. Innate immune responses triggered by MVA-HCV ΔC6L in vitro showed a downregulation of cytokines and chemokines compared to MVA-HCV whereas a similar profile of recruited immune cells was induced in vivo. Furthermore, MVA-HCV and MVA-HCV ΔC6L induced similar levels of high, broad and polyfunctional HCV- specific CD4+ and CD8+ T-cell adaptive and memory immune responses in vaccinated mice. This response was mainly mediated by CD8+ T-cells, which were primarily against p7, NS2 and NS3 HCV proteins. Antibodies against E2 were also induced. Conclusion: Deletion of C6L
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