Day 1 :
Keynote Forum
Dr. Fernando de la Calle
Head of Microbiology Dpt. PharmaMar. Madrid. Spain
Keynote: Marine Microbiome and Drug Discovery
Time : -
Biography:
Head of the Microbiology Dpt .at PharmaMar where he joined 30 years ago. He received his Ph.D. in Molecular Biology from UAM (Madrid) in 1998. He also completed an Executive-MBA in 2005. Co-author of the current hemisynthetic process for manufacturing the anticancer Yondelis®, the first European marine derived compound approved. He has been member of the International Advisory Group of ERA-MBT and BiodivERsA. Reviewer for H2020, FP7 and ERA-NET projects. He is the scientific contact person from PharmaMar for FP7/H2020 EU projects related to Marine Biodiversity & Biotechnology such as MAMBA, MaCuMBA, MicroB3 and INMARE.
Abstract:
Traditionally, higher plants and terrestrial microorganisms have proven to be the richest sources of natural drugs. However, we are living in a planet of oceans. Evolutionary development has equipped many marine organisms with the appropriate mechanisms to survive, developing exquisitely complex biological and chemical mechanisms for defence, attack, signalization and other still unknown purposes. These biological capabilities are clearly revealed by their ability to biosynthesize and release potent chemical weapons that are active per se. Such novel chemical structures often result in new modes of action and open up the potential of new ways to treat cancer and other diseases.
Early studies of the marine environment focused on natural products from invertebrates and tunicates have led to the isolation of several classes of bioactive natural products mainly small molecules such as polyketides and non-ribosomal peptides. However, there is an emerging a rational suspicion, based on marine sponge/tunicate metagenomics that these compounds originally isolated from metazoan organisms are in fact of bacterial origin. The cases of trabectedin (ET-743), didemnins, kahaladide, onnamides, bryostatin or dolastatin are some examples of the role of marine microbiome as producers of bioactive metabolites with application in oncology. Currently, genomic mining for polyketide synthases (PKS) and non ribosomal peptide synthetases (NRPS) improves the probability of success in drug discovery using marine microorganisms, both in isolated genomes and metagenomes.
Dr.de la Calle will discuss the state of the art in marine biotechnology applied to drug discovery under the experience of PharmaMar, spanish company focused in reseach, development and commercialization of marine drived drugs for the treatment of cancer.
- Water Microbiology
Location: Hall-1
Chair
Fernando de la Calle
Head of Microbiology Dpt. PharmaMar. Madrid. Spain
Session Introduction
Alla Nozhevnikova
Federal Research Center of Biotechnology of the Russian Academy of Sciences, Russia
Title: Mainstream partial nitrification-anammox process in domestic and municipal wastewater treatment in Russia
Biography:
Alla Nozhevnikova has completed her Doctoral Degree at Winogradsky Institute of Microbiology RAS and became Professor of Moscow State University in 1994. She is the Head of Laboratory of Microbiology of Anthropogenic habitats. She has published more than 150 papers in reputed journals and is a member of three Scientific Councils
Abstract:
Mainstream partial nitrification – anammox (PN/A) process is at the peak of interest for providing sustainable nitrogen removal from domestic and municipal wastewater. The interactions among key microbial groups: ammonium-oxidizing, nitrite-oxidizing, anammox, and heterotrophic bacteria are critically overviewed in a recent publication of Cao et al. (2017). Reactor design, flow pattern, carriers, oxic-anoxic conditions, pH, temperature are essential for effective cooperation of key mictobial groups. The principles of new PN/A technology and the design of full-scale prefabricated 100-1000 m3/day "BCH-ECOS" wastewater treatment plants (wwtps) were based on the results of microbiological studies and semi-pilot trials. New technology involves chemical enhanced pretreatment of the mainstream for significant part of carbon and phosphorus removal. Brush-shaped carrier is applied at all steps of biological treatment for microorganisms immobilization and biofilm formation. Wastewater with high ammonia concentration comes into the denitrificator (first step), which also receives water that is high in nitrate and nitrite by means of recirculation from the aeration tank (second step). N2 is produced by anammox- and heterotrophic denitrifying bacteria in denitrificator. Three genera and new specia of anammox-bacteria were detected in denutrificator biofilms. The quality of the treated water exceeds the requirements for discharging into water bodies. More than 20 "BCH-ECOS" plants are in operation in different regions of Russia since 2009. Wwtp “Megapolis” (5000-100000 m3/day) is developed based on the new technology for municipal wastewater treatment. All wwtps are isolated from the environment and keep constant temperature
Linda K. Medlin
Marine Biological Association of the UK, UK
Title: Advances in molecular tools for routine monitoring of toxic algae and pathogens in aquatic ecosystems
Biography:
Linda K. Medlin obtained her Ph.D. from Texas A&M University in 1983 and is currently a research fellow at the Marine Biological Association of the UK where she co-ordinated the FP7 EU project MIDTAL. During her employement at University of Paris and at Microbia Environnement, she directed 2 additional FP7 projects mAqua and SMS. All three of these projects involved the making of species specific probes for early warning systems for toxic algae and/or freshwater pathogens and applying them in a microarray or biosensor format and for making phylochips for the analysis of marine biodiversities using microarrays. She has published over 250 research papers including 35 chapters in books. She has participated in over 18 EU grants and many other national funding proposals from the UK and Germany
Abstract:
Microarrays are oligonucleotides applied to the surface of a glass slide in an ordered array. When rRNA sequences are used, these are called phylochips, which can identify organisms and is a relatively new, innovative microarray application. Phylochips can facilitate monitoring for any microorganism in any environment and visualize its changes in abundance over time for long-term records. We developed in three EU projects: a phylochip for the detection of toxic algae in marine waters and for freshwater pathogens in freshwater and tested them with environmental samples in 5 countries for the toxic algae and 6 countries for the freshwater pathogens. Water samples were filtered until they clogged or concentrated into one litre using a kidney dialysis filter, of which free filters are being distributed upon request. Total RNA was extracted using TriReagent, fluorescently labelled and hybridised to the phylochip. The pattern captured via fluorescent excitation in the microarray scanner is exported as an excel file and analyzed based on presence/absence of probe signals in a hierarchical fashion. For a species to be present, probes for higher taxa, viz., genus to kingdom must also be present. Where calibration curves have been made, then the microarray signal can be converted into cell numbers. In a fourth EU project microarray probes were transferred to a SHA coupled to an electrochemical and a colorimeter detection. The electrochemical detection was 16 fold higher than that obtained in earlier applications and the colorimetric detection was automated for a real time in-situ assay in a buoy
David Sartory
SWM Consulting Ltd., UK
Title: Evaluation of a rapid and simple MPN method for enumerating Legionella pneumophila from water
Biography:
David Sartory is an Independent Consultant, has been involved in Drinking and Environmental Water Microbiology for 40 years, with over 45 professional publications to his name. After qualifying in Microbiology from the University of Surrey, he joined Thames Water, before spending 8 years in South Africa, where he obtained an MSc in Limnology. Prior to his current role, he was Company Advisor for Severn Trent Water, where for 15 years he was involved in the development of improved methods for the microbiological assessment of water quality including the MLGA medium for E. coli and coliforms, and the Filta-Max system for Cryptosporidium analysis. He is the Chairman of the BSI Water Microbiology Committee and a member of several ISO working groups
Abstract:
Legionella pneumophila is an opportunistic pathogen of major concern, being the cause of severe respiratory diseases including Legionnaires’ Disease. Monitoring of at-risk water systems, particularly in hospitals and similar environments, is key to the control of this pathogen. This presentation will outline the results of a comparison of the performance of a novel and simple most probable number (MPN) method (Legiolert™/Quanti-Tray®) with the widelyused ISO 11731-2 membrane filtration method for the enumeration of L. pneumophila from water. Data from a multi-laboratory study which were analysed according to ISO 17994 showed that Legiolert™/Quanti-Tray® yielded on average higher counts of L. pneumophila, with results being available within 7 days compared to the 10 or more days required for the ISO method. The Legiolert™ medium also had a high specificity for L. pneumophila of 96.4%. This new method represents a significant improvement in the efficiency and simplicity of testing for the enumeration of L. pneumophila from water samples
- Infection and Immunity
Location: Hall-1
Session Introduction
John F. Alderete
Washington State University,USA
Title: A Recombinant Chimeric Protein Comprised of Immunogenic Epitopes of Metabolic Enzymes is Serodiagnostic Target for Trichomonas vaginalis Sexually Transmitted Infections
Biography:
Dr. Alderete received his PhD from The University of Kansas in 1978 at the age of 28. He did postdoctoral research at The University of North Carolina at Chapel Hill. He has published close to 140 scientific articles and has 63 book chapters, invited articles, and press releases. His work has been presented at 157 scientific conferences, and he has given seminars at 90 colleges and universities worldwide. He has served in NIH Study Sections, Boards of Scientific Counselors, and National Advisory Councils. He has been a member of several National Academy of Medicine panels
Abstract:
There is a need for a rapid, inexpensive and accurate serum antibody-based diagnostic with targets of high specificity for screening of large cohorts of women and men at risk of infection by Trichomonas vaginalis. This protist causes the number one non-viral sexually transmitted infection worldwide. The antigen-detection OSOM™ Trichomonas Rapid Test (Seskui Diagnostics) is a lateral flow, immunochromatographic Point-of-Care test that works only for women. During the course of our investigations of the T. vaginalis-host interactions, highly immunogenic proteins detected by sera of patients with trichomonosis but not uninfected controls were identified. Some of these immunogenic proteins include the metabolic enzymes fructose-1,6-bisphosphate aldolase (A), α-enolase (E), and glyceraldehyde-3-phosphate dehydrogenase (G). The epitopes of these proteins were characterized and found to have little or no sequence identity to other eukaryotes, yeasts, and microbial pathogens, including organisms that cause other STIs. We have constructed a new version of an earlier chimeric recombinant String-Of-Epitopes (SOE) protein consisting of 15-mer peptides of epitopes of A, E, and G (ImmunoTargets Therapy, 2013:2, 91-103). This composite protein called AEG::SOE2 was detected by ELISA with highly reactive sera of women and men but not control, negative serum lacking antibody to T. vaginalis. I believe that this approach lends itself to the creation of highly specific immunogenic targets for both detection of serum antibody in patients as well as for future subunit vaccines
Zhongtian Qi
Second Military Medical University, China
Title: Exosomal miRNAs derived from umbilical mesenchymal stem cells inhibit hepatitis C virus infection
Biography:
Zhongtian Qi has completed his MD from Second Military Medical University, China and Postdoctoral studies from New York University School of Medicine. He has been working on Clinical Microbiology for more than ten years, and published more than 75 papers in reputed journals
Abstract:
Background & Aim: Hepatitis C virus (HCV) infects approximately 3% of the world population. Though the development of direct acting antivirals (DAAs) have improved the sustainable virological response (SVR) rate in HCV patients, novel anti-HCV agents with higher efficacy and better tolerance are still urgently needed. Cell-based therapy, like exosomes, has become one of the most popular therapeutic methods in recent years.
Methods: Conditioned medium from umbilical mesenchymal stem cells (uMSC) is used to test its effect on HCV infection. Exosomes are further extracted and purified from the supernatants of uMSC (uMSC-Exo), and its anti-HCV activity is evaluated. Single cycle HCV pseudoparticles are applied to determine the effect of uMSC-Exo on HCV entry. Host cells are transfected with viral RNA or HCV replicon cells are utilized to detect the effect of uMSC-Exo on viral replication. The intracellular and extracellular infectivity are also evaluated to test the effect of uMSC-Exo on viral assembly and release. Proteinase K treatment assay is used to determine which components in uMSC-Exo are the functional substances. Small RNA sequencing is made with uMSC-Exo, and miRNAs with antiviral potency are identified. Function analysis is carried out by overexpression or knock down of relevant miRNAs, and their roles in inhibiting HCV infection are evaluated.
Results: uMSC inhibit HCV infection by paracrine, and uMSC-Exo are the main active constituents in this process. uMSC-Exo can enter Huh7 cells and reduce intracellular HCV RNA level as well as viral protein expression in infected cells. uMSC-Exo have no effect on viral entry, but suppress viral replication. Proteinase K treatment assay confirms that the RNA components are the active anti-HCV constituents in uMSC-Exo. Small RNA sequencing of uMSC-Exo indicates their miRNA expression profile. Among them, nine miRNAs are upregulated in the host cells after uMSC-Exo treatment. The functional analysis suggests four miRNAs (let-7f, miR-145, miR-199a and miR-221) play important roles in HCV infection. The inhibitory effect of uMSC-Exo is lost when the uMSC are transfected with the inhibitors of the four miRNAs.uMSC-Exo exhibit synergistic effect when combined with IFN or VX-950.
Conclusion: uMSC-Exo inhibits HCV infection by exosomal miRNAs (let-7f, miR-145, miR-199a and miR-221) with antiviral activity on viral replication. This work provides novel insights and possibility for developing anti-HCV therapy.
- Marine Microbiology and Ecology
Location: Hall-1
Session Introduction
Emiliya Kalamiyets
Belarus National Academy of Sciences, Belarus
Title: Contribution of Applied Microbiology into national economy of Belarus Republic
Biography:
Emiliya Kalamiyets has completed her PhD in microbiology in 1980 and Doctor of Biological Sciences thesis in 1999. She is General Director of Research and Production Association “Chemical synthesis and biotechnology”, Director of the Institute of Microbiology, Belarus National Academy of Sciences, Chairman of Belarussian Non-governmental Association of Microbiologists. She is a recognized expert in the field of microbial synthesis. Major trends of her research are genesis of biologically active compounds showing antimicrobial and entomopathogenic action, elaboration of biotechnologies for plant protection. She is an author of 341 publications, including 2 books, 15 patents
Abstract:
Significance of applied microbiology is determined by vital role of microorganisms in global natural cycles, technologies, medicine and ecology. Insitute of Microbiology, Belarus National Academy of Sciences, is the leading research institution of Belarus in the area of applied microbiology, specializing on evaluation of microbial biodiversity, physiology and biochemistry; genetic engeneering of microorganisms producing bioactive substances; development of biotechnologies for industry, agriculture, medicine, environmental protection.
Crucial aspect of microbiological processes is provision of superactive strains synthesizing bioactive substances. Both conventional selection methods and genetic engeneering are used to attain this goal. The Center of Analytical and Genetic-Engeneering Studies set up at the Institute is equipped with up-to-date instrumentation allowing to derive strains with defined properties engaged in elaboration of biotechnologies lying within the mainstream of global high-tech trends.
Molecular-genetic identification of microbial strains is carried out at Belarussian Collection of Non-pathogenic Microorganisms integrated into Institute structure and registered as a member of World Federation of Culture Collections. Apart from main collection stock, special DNA banks were founded for industrial strains, plant pathogens, xenobiotics destructors and bioterioration agents, to ensure preservation of genomes of promising biotechnological objects; early diagnostics of pathologies affecting forest and agricultural cultures; bioremediation of natural and technogenic environments; evaluation of divese materials bioresistance.
Technologies and production generated at the Insitute find outlet in numeric branches of national economy, including agriculture – biopesticides, microbial fertilizers, probiotics, biodesinfectants, biopreservatives, enzyme preparations; medicine – pharmaceutical technologies, diagnostic kits; industry – food additives, organic acids, alcohols, and makes a solid contribution into its progress
Biography:
Volpicella M. has completed his PhD and postdoctoral studies at the A. Moro University. From 2004 she is researcher in Molecular Biology at the Department of Biosciences, Biotechnology and Biopharmaceutics at the University of Bari. She has published 25 papers in reputed journal. Her recent research activity involves studies on bacteria adaptation to environmental stresses by genomics, transcriptomics and proteomics approaches., and also metagenomics of extreme environments
Abstract:
Backgrounds. Marine salterns are excellent sites for studying the dynamics of the prokaryotic biodiversity at increasing salt concentrations. Metagenomics offers the most direct approach for reliably assessing the microbial diversity including uncultivable prokaryotes.
Salterns of Margherita di Savoia (MdS) are located on the East-cost of South Italy. They are the largest salterns in Europe, with a yet unexplored microbiota composition.
Objectives. Defining the microbiota composition of the salterns of MdS in ponds with increasing salt concentrations.
Methods. eDNA purified from nine ponds with salt concentration in the 4.9-36% range was used for PCR amplification of the V5-V6 hypervariable region of the 16S rRNA gene. NGS of amplicon libraries was carried out by the Illumina MiSeq platform. Obtained reads were analyzed using the BioMas software for taxonomic classification.
Results and Conclusions. The microbiota composition of the MdS salterns resulted in a peculiar composition of prokaryotes, quantitatively different from that of other salterns of the Mediterranean area. For example, Archaea are absent at low salt concentrations (4.9-8.4%) and reach their highest concentrations (30-35%) in the high-salinity ponds. In similar ponds of the salterns of Santa Pola (Spain) their presence has been estimated around 90%. Conversely, in the high-salinity ponds of the MdS salterns, the Eubacteria Salinibacter genus is the most represented genus.
This study is of particular interest, not only to define the microbiota composition in different salt concentrations, but also for better addressing future functional metagenomics analysis aimed at the identification of biotechnological useful extremozymes
- Clinical Microbiology
Location: Hall-1
Session Introduction
Rasih Felek
Akdeniz University Hospital, Turkey
Title: Evaluation of Aspergillus immunochromatographic lateral flow device and galactomannan antigen test for the presence of invasive aspergillozis in patients with haematologic malignancies
Biography:
Rasih Felek has completed his MD at Hacettepe University School of Medicine and Postdoctoral studies from Atatürk University School of Medicine. He has published more than 15 papers in reputed journals
Abstract:
Diagnosis of Invasive Aspergillozis (IA) in patients with haematologic malignancies and under the risk of IA may be uncertain or delay because of nonspecific clinic presentation and difficult application techniques of conventional methods. Early diagnosis can provide initial antifungal therapy and prevent high mortality. In this study, we investigated the performance of Aspergillus lateral flow device and Galactomannan (GM) antigen test for the IA diagnosis of patients with febrile neutropenic episodes. The study was conducted in Akdeniz University Medical Faculty Hospital, Pediatric Haematology and Stem Cell Transplantation Units. 365 serum samples of 29 febrile neutropenic episodes belong to 21 patients with risk of IA were tested. According to the reference diagnosis of revised definitions of EORTC/MSG criteria, one patient was proven IA, 14 patients were probable IA, 2 patients were possible IA and 10 patients were under the risk of IA. While GM test had higher sensitivity, Aspergillus lateral flow device had higher specifity. Aspergillus lateral flow device is an easy to use assay with short hands-on time, but as long as combination of this assay with GM antigen test can lead increased sensitivity for diagnosis of IA
- Systems Biology & Bioinformatics/Plant pathology
Session Introduction
Olga Kudryavtseva
Lomonosov Moscow State University,Russian Federation
Title: Filamentous Fungus Podospora anserina: Morphological and Genomic Changes within 5 Years of Submerged Cultivation
Biography:
Olga Kudryavtseva graduated from Lomonosov Moscow State University (Faculty of Biology) in 2007, completed her PhD in 2011 with specialization in mycology, and continues to work over there as a Research Scientist. Also she conducted studies on fungal aging in collaboration with CNRS, Centre de Genetique Moleculaire, France. Today Dr. Kudryavtseva teaches a laboratory course on physiology of fungi and leads long-term study on the evolution of fungal model. Since 2013 she is a regular head of subsection of Mycology and Algology as a part of annual International Conference for Students and Young Scientists "Lomonosov"
Abstract:
Microbiological cultures can undergo microevolution in laboratory conditions. Comprehensive study of this process is important not only from fundamental point of view, but it could help successfully adapt natural and genetically modified strains to specific or stressful conditions of industrial cultivation, that may be quite different in comparison with their natural habitats. In the present study we observed genetic and phenotypic changes in ascomycetous fungus Podospora anserina grown at aerated liquid media more then 5 years by serial passages. P. anserina is coprophilous fungus living in nature on dung of herbivores. Two initial strains were derived from individual mononuclear ascospores and became original for eight independent subcultures. Experimental conditions influenced dramatically on fungal morphology as well as on sexual reproduction, but constantly support unlimited vegetative growth of mycelium. All subcultures being removed on agar medium at any time, as well as on dung agar, grow with the same rate as wild type strains. Method of next generation sequencing for whole genome analysis lets us revealed 142 point mutations fixed in experimental subcultures in total after 268 passages in liquid: 52 of them were identified as missense mutations, 8 as nonsense, and 10 as frameshifts. Moreover, we have observed some examples of parallel evolution, the most impressive of which was the gene coding GTPase subunit alpha that was subjected to mutation fixation independently in seven subcultures. Functional annotation of found mutations indicates that some of them might be adaptive. The reported study was supported by RFBR, research project No. 16-04-01845 a
Belén Guijarro
National Institute for Agriculture and Food Research, Spain
Title: Adaptive conditions and safety of the application of Penicillium frequentans as a biocontrol agent on stone fruit
Biography:
Belén Guijarro has completed her PhD from Politechnique University of Madrid and Postdoctoral studies from La Trobe Univeristy School of Botany of Melbourne. She is a Scientific Researcher of Plant Pathology in the Department of Plant Protection, Phytopathology Fungi Unit, at the National Institute for Agriculture and Food Research in Madrid. She has published several papers related to biocontrol of plant diseases
Abstract:
Many microorganism are with good antagonistic potential, however fail to be developed for practical use, due to inconsistent performance associated with field applications and often adverse environmental conditions. The EU emphasizes the necessity to know crop safety and evaluate the production of secondary metabolites on fruit surface together with their risk on human. Mycotoxins from Penicillium species can be a serious contaminant problem in storage fruit. Pf 909 is used as a BCA against brown rot in stone fruit caused by Monilinia spp. Penicillium frequentans (Pf909) efficacy is very dependent on its survival and persistence on flower and fruit surfaces, which depend on the climate. This study tries to achieve part of the registration requirements necessary for authorization of BCA Pf909 in EC countries. Identifying areas of research that still need to be addressed to demonstrate that Pf909 is sufficiently stable on field conditions and crop safe for registration purposes. This work is divided into three major sections: effect of physical factors on Pf909 development; the antagonist spectrum of resistance to commercial products use for conventional animal and human fungi control; and descharge mycotoxic production by Pf909 on fruit surface. Pf909 survive actively over a wide range of environmental conditions. The probability of risks to human and animal health is considered to be remote in terms of mycotoxic production, and there is commercial active material capable to inhibit fungus growth