4^th World Congress and Expo on Applied Microbiology November 29-30, 2017 Madrid, Spain

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

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

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

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

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.

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

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

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

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

Biography:

Sanin Musovic, a molecular microbiologist, completed both his PhD (2010) and 5-years postdoctoral studies at Danish Technical University. He is currently Specialist in Microbiology and Bioengineering at Danish Technological Institute, where he leads drinking water laboratory. He serves as: reviewer for 5 international journals, external/invited expert in EU commissions LIF Team, has over 30 publications (peer-reviewed journals and conference proceedings) etc.

Abstract:

Groundwater filtration by rapid sand filters (RSF) at waterworks is a widespread technology for drinking water production, which efficiently removes the naturally occurring pollutants (e.g., NH4) by a combination of chemical and biological (microbial) processes. During the past few decades, unnatural toxic substances, pesticides, have appeared in groundwater, which challenges the future stable supply of drinking water to consumers. The aim of current project was to develop an environmentally friendly biotechnology dealing with pesticides in ground- and drinking water. The biotechnology is based on natural microorganisms in RSF, possessing the abilities to degrade pesticides toward CO2 and H2O. These skills are frequently located on mobile genetic elements (MGE), which bacteria exchange among themselves. Application of natural isolates with pesticide-degrading genes on mobile genetic elements (MGE^P) would ensure transfer and permanent establishment of introduced MGE^P among the native microbial communities. Hence, the Trojan Horse biotechnology`s omits the well-known limitations of low-to-none survival of exogenously introduced pollutant degrading bacteria in the environment.

Results shown that prolonged (½ year) exposure of sand material to investigated pesticides enhanced the fraction of pesticide-degrading bacteria in enrichments, ensuring isolation. Pesticides effected the structure of microbial communities, reducing abundance of e.g. Alpha Proteobacteria and Nitrospira sp by MCPP, or stimulation Springobacteriia sp. by glyphosate.

Suitability of Trojan Horse concept was confirmed by experiments combining plasmid-borne reporter (gfp) gene and flow-cytometric approach, revealing dominance of Alpha-, Beta- and Gamma Proteobacteria among native sand filter bacteria that acquired investigated plasmid (MGE). (Up to 250 words)

Biography:

Mihai Nita-Lazar has a wide expertise in the fields of microbiology, molecular biology and biochemistry mainly developed in prestigious laboratories such as the Institute of Microbiology, ETH, Zurich, Switzerland, the Department of Biochemistry and Cellular Biology of Stony Brook University, New York, SUA, the Department of Cellular and Molecular Biology of Medical University Center from Boston, Massachusetts, SUA and the Department of Microbiology, University of Medicine, Baltimore, Maryland, SUA. He is currently working as principal investigator of Bioassay-Biological Analysis Laboratory of the National R&D Institute for Industrial Ecology-ECOIND. He has more than 40 ISI papers, 4 books/chapters, h-index 10.

Abstract:

The waterborne pathogenic bacteria, especially the enteric bacteria of human fecal origin have become currently a global public health issue. The detection and quantification of drinking water microorganisms are an essential part of any quality control or water safety management plan interconnected to enteric bacterial pathogens such as Salmonella spp., Shigella spp., Fibrio cholerae or to nonfecal bacterial pathogens such as Legionella spp. The standard methods of detecting waterborne pathogenic bacteria are time-consuming due to the growing step in a specific culture media, followed by isolation, microbiological and/or serological identification and in some cases followed by subspecific characterization.

This study aimed to develop a faster, powerful, more sensitive and reproducible diagnostic tool to monitor a specific pathogen contamination in drinking water by specific antibody-antigen interactions. Three pathogenic bacteria such as Pseudomonas aeruginosa, Escherichia coli and Legionella sp. were detected by immunofluoresce technique with fluorochome tagged antibodies. Our results showed a good specificity of the antibodies in a very complex bacterial mixt as well as a starting detection level from 1 bacteria / ml.

Overall, these technique proved to be a reliable one, time-effective and sensitive for diagnosis and prevention of drinking water quality and waterborne bacterial disease.

Biography:

Sanin Musovic, a molecular microbiologist, completed both his PhD (2010) and 5-years postdoctoral studies at Danish Technical University. He is currently Specialist in Microbiology and Bioengineering at Danish Technological Institute, where he leads drinking water laboratory. He serves as: reviewer for 5 international journals, external/invited expert in EU commissions LIF Team, has over 30 publications (peer-reviewed journals and conference proceedings) etc.

Abstract:

Biological rapid sand filters (RSF) are commonly used for drinking water production at waterworks in Denmark, Europe and worldwide. RSF is a place where natural groundwater pollutants (e.g. ammonium) are removed. The native microorganisms in RSFs have an important role in removal of pollutants, where a sudden change in e.g. ammonium removal suggests a misbalance in microbial community. The aim of current work was to get a deeper insight in the physiological needs of main microbial groups at RSF to micronutrients (trace elements). Waterworks frequently receive groundwater with different chemical properties (incl. micronutrients) from distant water-wells, which allows creation of well-defined inlet-water blend.qPCR based approach revealed that sand filters were microbial rich environments, harboring ca. 3 billion bacteria per gram sand. The ammonium oxidizing bacteria (AOB) stayed for 1-3 % of bacterial population. AOB were up to 50-times more abundant in pre-filter units than in respective after-filter units. The abundance of ammonium oxidizing archaea was trivially low, suggesting that AOB stayed for full ammonium removal in RSF. Both the laboratory column-experiments and full-scale trials at waterworks suggested that certain sand filters at a waterworks naturally possessed an extra NH4- removal capacity. A fine blending of inlet water from chosen water-wells, focusing on copper and nickel at low (0.1 – 1.0 ug/L) concentrations, shown a high (>50%) and persistent stimulating effect on NH4 removal by ammonium oxidizing bacteria. A sudden stimulating-effect of nickel and cupper appeared to be product of increased catabolic abilities of the existing AOB cells.

Biography:

Catalina Stoica has completed her PhD in 2016 from University of Bucharest, Ecological Systems and Sustainability Department, Faculty of Biology in Romania. She is currently working as a research scientist in Laboratory of Bioassay-Biological Analysis of the National R&D Institute for Industrial Ecology-ECOIND, the only institute in Romania that displays a global approach on industrial ecology and environmental issues. She has published more than 18 papers both as first author and co-author in international journals as well as three co-authored book chapters.

Abstract:

The human population increase led to agricultural and farming development, which is limited by its land availability. Now a days, a major direction is based on the production efficiency and on the economy of size. The economy of size aims to lower the average cost per unit of production as the production increased. In our study, we used a very abundant and economically efficient nutrient to reduce the feeding costs, but to keep at least the same level of eggs and meat poultry production.The new nutrient formula was based on cellulose fibres, combined to a cellulosic enzymatic food supplement. Our in vitro results showed that by adding a cellulase based compound to the new feeding nutrient the cellulose content decreased, so the poultry could digest the new and economically efficient diet without any duodenal negative impact. In vitro enzyme – substrate assays were performed on 14 cellulose based nutrients (substrates) and 2 cellulase food supplements (enzymes). The results showed a 50% efficiency of cellulose degradation rate during 1h for an 1:1 enzyme-substrate ratio, especially for the Biozym M6000 food supplement. Moreover, the nutrients and the food supliments were microbiologically tested for the presence of Escherichia coli and Salmonella sp. The results showed the absence of Salmonella sp., but the presence of Escherichia coli in a density ranged between 33 and 1609 CFU/ml. Overall, the results showed an economical viable solution for poultry farms based on a rich cellulose based nutrients supplemented with cellulase food supplement.

Biography:

Dr Ajayi obtained his Ph.D. (Microbiology) degree from the University of Ibadan, Ibadan, Nigeria in 2005. I had diversified working and research experience in the field of Environmental Microbiology and Antimicrobials. He published over 50 research articles in learned journals worldwide. His research activity progresses with surveillance of antibiotic resistant reservoirs during Postdoctoral fellowship program in the Department of Biochemistry and Microbiology, University of Fort Hare, Private Bag X1314, Alice 5700, South Africa sponsored by Govan Mbeki Research and Development Center of the institution. He is currently the acting Head of Department of Microbiology, Adekunle Ajasin University, Akungba-Akoko, Nigeria.        

Abstract:

Various types of bacterial isolates were obtained from commonly consumed food sources, viz rice and beans in some Akoko communities, Nigeria and the world at large. The bacterial isolates were identified by standard microbiological methods. Proximate analysis was carried out according to the procedure of association of official Analytical Chemists to determine amount of components such as fat content, protein content, moisture content and ash content in food samples. Bacterial isolates encountered during the study includes Clostridium spp. (4), Vibrio cholera (1), Bacillus spp. (3), Escherichia coli (2), Staphylococcus aureus (2), Shigella spp. (2), Pseudomonas spp. (2), Streptococcus spp. (2), and Micrococcus spp. (6). Proximate analysis determined, shows the values of food components (in percentages, %) for rice and beans as, Moisture content (11.76, 13.96), Ash content (0.68, 2.52), Crude Fat (3.00, 6.65), Crude protein (15.67, 21.58) Crude fibre (1.44, 0.32) and Carbohydrate content (67.45, 54.97). This study helps to investigate field samplings of the staple food studied in that microbes isolated from the commonly consumed food (rice and beans) in this part of Nigeria and world at large, were used as a measure to determine level of contamination and possible microflora that may inhabit these food sources for possible industrial applications. Similarly, the proximate analysis done helps to determine dietary value as well as food safety. Hence, these food sources should be properly handled coupled with appropriate storage based on their nutritional value and liability of contamination at difference stages of their processing from the raw source. 

Biography:

Abstract:

Phenylpropenes (PPs) are volatile hydrophobic phytochemicals generally characterized by their potent antibacterial activities. The modes of antibacterial actions of these molecules differ from those of conventional antibiotics, which suggests their usage may help overcome antibiotic resistance mechanisms.The efficacy of allylic/propenylic PP isomers (eugenol/isoeugenol and estragole/anethole) was evaluated by macrodilution methods allowing to determine IC50s, MICs and MBCs against reference strains, Escherichia coli ATCC 25922 and Staphylococcus epidermidis ATCC 14990. Results indicate that all PPs were efficient against both bacteria in low millimolar levels, which makes them considerably less potent than common antibiotics. Allylic/propenylic isomers presented very similar growth inhibition patterns, indicating that the position of the double bond in the propenyl chain has low influence on PPs efficacy against the studied strains. For both strains, eugenol and isoeugenol were two to three times less effective than anethole and estragole, indicating distinctive modes of actions between these two pairs of isomers. Eugenol and isoeugenol would particularly involve their hydroxyl group to elicit antibacterial activity. Lacking a free hydroxyl group, the mode of action of anethole and estragole may be rather related to their higher lipophilic character. Interestingly, Gram-negative E. coli, showed to be more susceptible to all PPs than the Gram-positive S. epidermidis strain, probably due to the easier permeation of PPs across the hydrophobic cell emvelope structures of E. coli. Using high-dose PPs (low millimiolar levels) in vivo may not be an optimal strategy, however combination of phytochemicals presenting different mechanisms of action may allow reduction of efficient doses.

Biography:

Ayman K. El Essawy has Ph.D in Microbiology, (Ain Shams University, Egypt), a diploma in Hospital Infection Control (Claude Bernard-Lyone 1 University, France) and a Diploma in Biostatistics (Ain Shams University, Egypt). He is fellow of Microbiology at Ain Shams University. He worked at Al Azhar University & American Naval Medical Research Unit No.3 (NAMRU-3) and Ain Shams University Genetic Engineering/biotechnology center. He is publishing mainly in the field of Microbiology and Molecular Microbiology. He is particularly interested in the study of bacterial resistance to antibiotics.  

Abstract:

The authors previously, achieved a high yield of exopolysaccharide production from a marine Klebsiella sp. The aim of the present study is determination of this exopolysaccharide composition and evaluation of its biological activities. The monosaccharide moieties of this exopolysaccharide were determined by complete hydrolysis, qualitative and quantitative paper chromatography of the hydrolysis product. The monosaccharide composition of this exopolysaccharide was galactose 16%, fructose 20%, glucose 32%, fucose 22%, and uronic acid 10%. The biological activities of the native and modified sulfated bacterial exopolysaccharide were investigated. No anticoagulation or fibrinolytic activities were recorded for the sulfated exopolysaccharide, but the native form of exopolysaccharide showed both activities. The native exopolysaccharide also showed higher antibacterial activity against S. aureus and E. coli but both the native and sulfated form had no antifungal activity against Candida albicans. MIC of the native and sulfated exopolysaccharide that sustain the highest bacterial inhibition zone was 15 mg/dl. No prebiotic activities were recorded for both. In conclusion, this biopolymer has significant biological activities and it has two rare monosaccharide moieties with very significant value namely fucose and uronic acid that require further adaptation for practical and industrial application.

Biography:

Iris Yedidia is the Head of the Ornamental Plants and Agricultural Biotechnology Dep. at the Agricultural Research Organization (ARO), Volcani center, Israel. Since 2004, a scientist and a team leader focusing on applying biotechnological tools to improve ornamental bulbs resistance to bacterial pathogens and studying the involvement of natural products in bacterial pathogenesis.

Abstract:

Plants produce diverse array of low molecular-mass compounds, with more than 8,000 phenolics. Phenolic compounds play a pivotal role in plant defence and fitness. Here, we provide an example for interference of plant small molecules (phenolics) with bacterial virulence, via inhibition of the quorum sensing (QS) machinery of the soft rot bacterium P. carotovorum.

Interestingly, biofilm formation and exoenzymes activity were significantly impaired, at compounds concentrations that did not affect bacterial cell growth or membrane integrity. Since biofilm production and exoenzymes are  virulence determinants known to be under the strict control of QS, the effect of specific molecules on QS was studied. Common volatile and soluble  phenolic compounds were tested for their influence on the expression of central QS system and QS controlled genes. The N-acyl-homoserine lactone (AHL) reporter strains (CV026 and pSB401) demonstrated a prominent reduction in the level of QS signal molecules accumulation, following exposure to the compounds. Moreover, infection capabilities were strongly impaired on potato, cabbage and calla-lily; but fully recovered upon external application of AHL. To further confirm interaction of the plant compounds with bacterial QS targets, drug discovery tools were occupied using the SCHRODINGER^® molecular docking suit.  Pectobacterium central QS proeins  ExpI/ExpR were used as targets for salycilic acid and carvacrol (plant phenolics). Finally uisng isothermal calorimetry (ITC), SA and CAR were diectly bound to cloned and purified ExpI, by this, experimentally supporting the computational docking results. True binding of plant derived phenolics to bacterial QS synthase protein as a target was demonstrated.  The importance of plant-bacteria chemical signalling shown here, supports a rising field of research endeavouring interkingdom communication between plants and bacterial pathogens.

Biography:

Dr. Min-Kyu Kwak holds a Ph.D. in School of Biological Sciences, and Institute of Microbiology, Seoul National University, Republic of Korea. He has completed postdoctoral studies from Seoul National University. He has been appointed as a professor in Sungkul University in Korea and as a visiting professor in University of Malaysia in Sabah. He is an editorial board member of Academy of Annals of Clinical Immunology and Microbiology and an organizer of the International Congress on Science and Technology. He has published/prepared more than 22 papers in international and national journals and proceedings, as first author and corresponding author. He also presented 13 kinds of Korea and international patents regarding cyclic dipeptides inhibitory to against multidrug-resistant bacteria, pathogenic fungi, and influenza A virus.    
 

Abstract:

High methylglyoxal content disrupts cell physiology, but mammals have scavengers to prevent glycolytic and mitochondrial dysfunctions. In yeast, methylglyoxal accumulation triggers methylglyoxal-oxidizing alcohol dehydrogenase (Adh1) activity. While methylglyoxal reductases and glyoxalases have been well studied in prokaryotes and eukaryotes, experimental evidence for methylglyoxal dehydrogenase (Mgd) and other catalytic activities of this enzyme affecting glycolysis and the tricarboxylic acid cycle is lacking. A glycine-rich cytoplasmic Mgd protein, designated as Mgd1/Grp2, was isolated from glutathione-depleted Candida albicans. The effects of Mgd1/Grp2 activities on metabolic pathophysiology were investigated using knockout and overexpression mutants. We measured glutathione-(in)dependent metabolite contents and metabolic effects, including viability, oxygen consumption, ADH1 transcripts, and glutathione reductase and α-ketoglutarate dehydrogenase activities in the mutants. Based on the findings, methylglyoxal-oxidizing proteins were monitored to determine effects of MGD1/GRP2 disruption on methylglyoxal-scavenging traits during glutathione deprivation. Methylglyoxal-oxidizing NAD(H)-linked Mgd1/Grp2 was found solely in glutathione auxotrophs, and it catalyzed the reduction of both methylglyoxal and pyruvate. MGD1/GRP2 disruptants showed growth defects, cell-cycle arrest, and methylglyoxal and pyruvate accumulation with mitochondrial impairment, regardless of ADH1 compensation. Other methylglyoxal-oxidizing enzymes were identified as key glycolytic enzymes with enhanced activity and transcription in MGD1/GRP2 disruptants, irrespective of glutathione content. Failure of methylglyoxal and pyruvate dissimilation by Mgd1/Grp2 deficiency leads to poor glutathione-dependent redox regulation despite compensation by Adh1. This is the first report that multifunctional Mgd activities contribute to scavenging methylglyoxal and pyruvate to maintain metabolic homeostasis and the redox pool via glycolytic enzymes and Adh1 expression.

Biography:

Sergio Giannattasio has completed his studies in Chemistry from Universtity of Bari, Italy in 1982. He is Senior Scientist at the Institute of Biomebranes, Bioenergetics and Molecular Biotechnlogies of CNR in Bari. In 2014 he has been awarded with the qualification as full professor in Applied Biology. He has published 66 papers in reputed JCR journals with 1586 citations and h-index 24 (Google Scholar) and has been serving as an editorial board member of repute. His research interests are mitochondria-to-nucleus cross-talk in cell stress response and the role in tumorigenesis of oncosupressor BRCA2 using yeast Saccharomyces cerevisiae as a model.            

Abstract:

More than 70% of prostate cancer (PCa) patients display loss or reduction of BRCA2 protein, an oncosupressor involved in DNA repair through homologous recombination. We have shown that loss of BRCA2 confers cells resistance to anoikis, a peculiar form of programmed cell death (PCD) necessary for cancer cells to colonize distal sites during the metastatic process. This PCD-promoting function of BRCA2 is conserved in the yeast Saccharomyces cerevisiae. Patients with metastatic PCa display only temporary disease control following androgen deprivation but they eventually develop disease progression to virtually incurable castration-resistant prostate cancer (CRPC). We used yeast expressing BRCA2, normal prostate cell and a CRPC cell line model for a pre-clinical toxicity screening of 6-thioguanine (6-TG) and six 6-TG analogues. We found that 6-TG decreased proliferation in yeast preferably in the presence of BRCA2 and normal prostate cells without inducing cell death. Strikingly, 6-TG not only impaired cell proliferation but also induced significant cell death in CRPC cells by activating a PCD program. Silencing of BRCA2 expression increased CPRC cell sensitivity to 6-TG-induced cell death. 2,6-dithiopurine (2,6DTP) strongly decreased proliferation and induced cell death in yeast independently of BRCA2. 2,6DTP and 2-amino-6-bromopurine selectively induced PCD in BRCA2-expressing CPRC cells. These results suggest the potential use of 6-TG and its analogues for the treatment of CRPC alone or in combination with taxanes, chemotherapeutic drugs approved for CRPC treatment. Acknowledgments: This work was supported by FCRP “Identificazione di molecole attive per lo sviluppo di nuovi farmaci anti-tumorali contro il carcinoma di prostata”.

Biography:

Xiaoling is a full professor of School of Mechanical Engineering at USTB, and a visiting scholar of School of Engineering and Applied Science at Harvard University. She received her BS and MS degrees in Solid Mechanics from Beijing Institute of Technology in 2000 and 2003, and a PhD degree in mechanical engineering from The Hong Kong University of Science and Technology in 2006. She was an postdoctoral fellow at The Hong Kong University from 2006 to 2007, and moved to USTB in 2008. she was a visiting scholar in the Northwestern University of USA. Her research interests focus on Biomechanics, Phase Transitions in bio system, Bacterial biofilms.

Abstract:

We develop an optical imaging technique for spatially and temporally tracking biofilm growth and the distribution of the main phenotypes of a Bacillus subtilis strain with a triple-fluorescent reporter for motility, matrix production, and sporulation. We develop a calibration procedure for determining the biofilm thickness from the transmission images, which is based on Beer-Lambert’s law and involves cross-sectioning of biofilms. To obtain the phenotype distribution, we assume a linear relationship between the number of cells and their fluorescence and determine the best combination of calibration coefficients that matches the total number of cells for all three phenotypes and with the total number of cells from the transmission images. Based on this analysis, we resolve the composition of the biofilm in terms of motile, matrix-producing, sporulating cells and low-fluorescent materials which includes matrix and cells that are dead or have low fluorescent gene expression. We take advantage of the circular growth to make kymograph plots of all three phenotypes and the dominant phenotype in terms of radial distance and time. To visualize the nonlocal character of biofilm growth, we also make kymographs using the local colonization time. Our technique is suitable for real-time, noninvasive, quantitative studies of the growth and phenotype distribution of biofilms which are either exposed to different conditions such as biocides, nutrient depletion, dehydration, or waste accumulation.

Biography:

Grettel Aviles is currently a graduate student in agricultural science at the Universidad Veracruzana, México.
 

Abstract:

The toxicity of nickel and vanadium, potentially toxic elements (PTE), in higher organisms has been widely documented because of its impact on crops or human health, however, toxicity in microorganisms has been poorly studied. The objective of this work is to determine the toxicity of nickel and vanadium in the growth of four strains of plant growth promoting microorganisms. Strains of Candida orthopsilosis B20, Serratia marcescens C753, Pseudomona tolasii P61 and Rhizobium tropici Ciat889 were inoculated into a minimal salt medium at pH of 6.5 at increasing concentrations of VOSO4 (0, 15, 30 and 45 mg · L-1) and NiSO 4 (0, 25, 50 and 75 mg · L-1). They were shaken at 140 rpm at 28 ± 2 ° C for 5 days. Their growth kinetics were measured by the turbidity method in a microplate spectrophotometer. Every 12 h, 200 μl of each sample was taken in triplicate and read at 600 nm, until 120 h. The data were processed by ANOVA, and Tukey's mean comparison test (α = 0.05), with the SAS program for Windows 9.0. The growth of C. orthopsilosis B20 was severely affected by the presence of the two metals, R. tropici Ciat889 was highly sensitive to nickel in all concentrations, while in vanadium, in its lower concentration, it promoted its growth. Both strains of S. marcescens C753 and P. tolasii P61 showed tolerance to PTE and their growth was promoted in low concentrations.

Biography:

Nadeem Hafeez from Lahore, Pakistan. Currently, I am working as Researcher at Center for Excellence in Molecular Biology (CEMB), Lahore, Pakistan. I have expertise in gene editing, cloning and plant transformation; I have developed a first triple gene genetically engineered cotton variety of Pakistan, which confers significant resistance against biotic and abiotic stresses. I have utilized E.coli and yeast expression models to screen mutated genes before transformation in plants. I am currently doing a research project in abiotic tolerance of cotton to learn more about reverse genetics. In this project we are using CRISPR cas9 genome editing to study genes associated with abiotic stress tolerance.

Abstract:

Cotton is a cash crop of Pakistan and Gossypium arboreum is a locally cultivated variety, which has considerable resistance against various biotic and abiotic stresses. This variety of cotton is considered as good reservoir of stress tolerance genes, while based on EST data mostly of its genes are uncharacterized. Universal stress protein-2 (USP-2) gene was identified in 15 days drought stressed leaves of G.arboreum-FDH-171. Full length of this gene was mutated at three different (M₁usp-2, M₂usp-2, M₃usp-2) positions (fig: 1) in three separate clones in E.coli-uspABC-mutant and Pichia pastoris-gs115 strains for its functional validation under various abiotic stress treatments (NaCl 800mM, PEG 8%, Heat 37-45⁰C, Cold 4⁰C). The expression of 1^st mutant (M₁usp-2) was noted as 8.3fold under NaCl stress and 9.7fold under PEG stress treatments, recombinant cells showed higher growth up to 10^-5 dilution in spot assay as compared to control and other genes. The 2^nd mutant form of USP-2 was expressed on induction but it was failed to initiate stress tolerant mechanism in both organisms. No significant difference was noted in between 3^rd mutant form and wild type USP-2. However, all mutant forms showed little tolerance against heat and cold stresses. The results of this study showed that activity of USP-2 was enhanced in M₁usp-2 by enhancing its ATP binding capacity at 2X but wipe out in M₂usp-2 with zero ATP-binding ability and 4X enhanced CMP capacity has no effect on activity of M3-usp-2. In silico analysis showed that 1^st and 3^rd mutant forms of USP-2 may directly involved in stress adaptive mechanism or it might be function as a signaling molecule to initiate stress mechanism.