14^th International Conference on Microbial Interactions & Microbial Ecology August 19-20, 2019 , Mercure Hotel Wien Westbahnhof ,Vienna, Austria

Guliko Dvali has completed her PhD in Microbiology from Ivane Javajishvili Tbilisi State University. She is the Head of mcrobilogy research group in Georgian Technical University, Biotechnology Center. She has published more than 20 papers in reputed journals and has been a great experience in plant miocrobiology field.

Study of the biopreparation influence on pathogens, which causes the disease of soil micro-flora and plant, is one of the main issues for environmental recovery and ecologically clean product. The purpose of the research was to study the influence of the pathogen Fusarium causing the disease of tomato root and the study of the influence of biopreparation “Phytocatena” (Pseudomonas fluorenscens), produced by the Plant Biological protection Center in Georgia, on the pathogen. The tomato variety “Slivka” has been studied for the experiment. Microbiological analysis was conducted by M. Wozniakovskaya’s method. The cultivation of microbes was occurred in potato, Chapek, Suslo’s food areas, with various suspensions (1:10, 1:100, 1:1000). The total quantity of pathogenic fungi was estimated by thousands of 1 gram on dry ground. As a result of research, the total quantity of pathogenic fungi in the untreatment soil was 400 thousand. The dominant pathogen was Fusarium (205 thousand). Soil and tomato root system was processed by the 2% solution of bio-preparation "Phytocatena" before the planting, and then by 2-4 times with 15-20 days interval. It has been determined, that the total quantity of pathogenic fungi in untreatment soil was 261 thousand (140 thousand on plant’s roots,121 thousand in the rhizosphere). The total quantity of Fusarium was 120 thousand (65 thousand on plant’s roots, 55 thousand in rhizosphere). From samples collected in the phase of tomato blossom, it has been found, that after the treat by the biopreparation "Phytocatena", the total quantity of Fusarium was significantly reduced-20 thousand (11 thousand on plant’s roots, 9 thousand in the rhizosphere). The total quantity of pathogenic fungi was also reduced by 49 thousand (25 thousand on the plant’s root, 24,000 in the rhizosphere).

Tamar Shamatava has completed her PhD at St. Andrew the First Called Georgian University of the Patriarchate of Georgia from 2010-2015. She is the Senior Scientist at Georgian Technical University Biotechnology Center. She has published more than 17 papers in reputed journals. She has a great experience in agriculture and biotechnology field

Background &Aims: The loss causing of pathogenic microbiological diseases during storage has a high economic impact. The research aim was to study effect of eucalyptus extract with combination calcium chloride on microflora of grapevine during storage. Materials & Methods: Two grapevine varieties were selected for study: Alphonse Levallée and Italia. Two combinations of eucalyptus extract and calcium chloride were selected for experiment: I.1 % CaCl2 and 2% eucalyptus extract II. 2% CaCl2 and 1% eucalyptus extract III. Control-untreatment grapevine. Treatment and control both were stored storage refrigerator -POLAIR Standard (temperature-0-10C, humidity- 85-90%). Findings: Pathogenic clear cultures were extracted from infected grapevine during storage (60-120 day). It was revealed that Botrytis cinerea and Penicilium expansum were two major infected agent which causing microbiological disease of grapevine varieties Alphonso levalee and Italia. Characterization and identification of fungi carried out using 40X-2500X professional infinity Trinocular Compoud Microscope (SKU:T690C). As a result showed the loss caused from phytopatohenic fungi were different-Control for grapevine varieties Italia with Botrytis cinerea was-55.3% and Penicilium expansum-37.6%. For Alphonso Levalée by Botrytis cinerea-54.1% and Penicilium expansum-35.2%. The best result for grapevine varieties Italia was showed 2% CaCl2 and 1% eucalyptus extract, in this case loss causing by Botrytis cinerea-42. 8%, and Penicilium expansum- 32.4%, but inhibition effect caused from Botrytis cinerea for Alphonso Levalee was-45.1% and Penicilium expansum-30.4%. Conclusions: Thus, the combination of 2% CaCl2 and 1% eucalyptus extract had inhibition influence on developments of Botrytis cinerea and Penicilium expansum, especially on Botrytis cinerea.

Vitalij Kolodynskij is a phD student of Environmental protection in Vilnius Gediminas technical university.

Bioreactor-special device is used for biogas production from various organic materials under anaerobic conditions. In this research, a batch bioreactor with a mechanical mixer was used for biogas production from sewage sludge and chicken manure bioloadings. The process of anaerobic digestion was mesophilic (35oC). Produced biogas was stored in a gasholder and the concentration of its components was measured with INCA 4000 biogas analyser. Also, a specific additive (pine wood biochar) was applied to prepare bioloadings. The application of wood biochar in bioloading increases the CH4 concentration in the produced gas by 6-7%. The highest concentrations of CH4 were found in biogas produced during the decomposition of sewage sludge bioloadings. The maximum CH4 reached 77.4%. Studies have shown that the application of biochar in bioloadings also reduces average CO2 and H2S concentrations in biogas.

Ting Zhao is a Research Assistant of Immunology at Chinese Academy of Medical Sciences and Peking Union Medical College. She is a Biotechnology Undergraduate Major by training at Yunnan University. She holds a Master’s degree in Immunology and earned her Doctorate in Immunology at Chinese Academy of Medical Sciences and Peking Union Medical College (China). She is especially interested in the mechanism of virus infection and the development of new vaccine. Her previous work is focused on Enterovirus 71 and development of Enterovirus 71 vaccine, especially about the neonatal rhesus macaque model of Enterovirus 71 infection model. Currently, her focus is on the comparative study of poliovirus immunization strategies in China during a transitional period.

Identification of mechanisms that improve mucosal antibody response to poliovirus vaccine and limit poliovirus replication is crucial for achieving the aim of global polio eradication. Since 2016, trivalent OPV using in developing areas has been required to switch to the bivalent OPV in association with one or two dose of IPV administered. Compared with IPV, oral poliovirus vaccine (OPV) not only induces serum antibodies against paralysis, but also induces a high level of intestinal immunity to prevent fecal-oral spread of poliovirus, which is the primary transmission route. Thus, after cessation of poliovirus type 2 (PV2), several studies indicated that mucosal neutralizing activity to PV2 decreased and virus shedding increased after a one-dose challenge with monovalent OPV (mOPV) type 2 in the bivalent OPV (bOPV) and bOPV-IPV groups compared with the tOPV group. In this study, we examined the levels of mucosal immunity to poliovirus in China by measuring the IgA antibody levels in stool samples collected from infants after sequential vaccination combining inactivated polio vaccine (IPV) with OPV. And then the pre-vaccination bacterial microbiota was identified using 16s ribosomal RNA sequencing. Our study shows that the new routine vaccination schedule reduced the induction probability of anti-poliovirus type 2 IgA as a result of the lack of type 2 components in bOPV. These results also detect Clostridia enteropathogens that could be linked with the decreased IgA conversion rate to poliovirus vaccine, which is a biomarker of mucosal antibody response. In addition to, rich diversity of the intestinal microbiota was a disadvantage in terms of the mucosal antibody response to OPV vaccine

Ahmet Volkan Kurtoglu is pursuing his Medicine at Bezmialem Vakif University. He has a special interest on infection diseases and predatory bacterias, which is an effective alternative to antibiotic therapies for challenging with drug resistance. He has worked on predatory bacterias for two years both in Bezmialem Vakif University clinics and Beykoz Institute of Life Sciences and Biotechnology (BILSAB) under the mentorship of Prof. Dr. Mehmet Ziya Doymaz. His methodology of the experiment was based on the method created by Prof. Elizabeth Sockett. The study indicates the importance of predatory bacterias and is one of the projects done in this restricted area.

Introduction: Misuse of antibiotics has caused increase in drug-resistance among bacteria and this forced the researchers to find new strategies and tools to encounter the growing threat of public health at a global scale. And predatory bacteria are one of these options. Bdellovibrio bacteriovorus and Agromyces ramosus species are two examples of such predatory bacteria with traits proven on several clinical pathogens. Aims: The aim of the project was to assess the predatory abilities of predator bacteria Agromyces ramosus and Bdellovibrio bacteriovorus, against clinical pathogens, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa, isolated in Bezmialem Clinical Microbiology Laboratories. Method: Assay for B.bacteriovorus: For creating the control group, each pathogen K. pneumoniae, A. baumannii and P. aeruginosa, was cultivated one night before, and prepared in falcon tubes at OD (600)=1 in 10 ml HepesCa++Mg++ and incubated for 2 days. Bacteria were counted as CFU on LB agars before and after incubation by serial dilutions. Likewise, one sample of each pathogen was prepared in tubes at OD (600=1) in 10 ml HepesCa++Mg++. Each tube was mixed with 300μl of Bdellovibrio bacteriovorus which had been prepared in HepesCa++Mg++ with using E.coli as prey. Prepared B. bacteriovorus co-cultures also incubated for 2 days. After 2 days, pathogen bacteria numbers were counted as CFU on LB agars before and after incubation by serial dilutions. Assay for A.ramosus: For control group, one sample of each pathogen, K. pneumoniae, A. baumannii and P. aeruginosa, were cultivated in for 24 hr before, and prepared in falcon tubes at McFarland=0.5 in 10 ml Brain Heart Infusion buffer (BHI), and incubated for 2 days. Bacteria were counted as CFU on BHI agars before and after incubation by serial dilutions. Similarly, each pathogen was diluted to McFarland=0.5 in 10 ml BHI buffer and mixed with 2ml A. ramosus diluted to McFarland=0.5 in BHI buffer. A. ramosus co-cultures were also incubated for 2 days. After 2 days, the pathogens were counted as CFU on BHI agars before and after incubation by serial dilutions. Result: B. bacteriovorus reduced all tested pathogen cell densities significantly (p<0.05). A. ramosus was significantly effective against Acinetobacter baumannii isolates (p<0.005). However, no such effect was noticed against Klebsiella pneumoniae and Pseudomonas aeruginosa. Conclusion: According to results, B. bacteriovorus could be considered as a predatory agent against Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa alternative to antibiotics. Further research on the applicability of these predatory agents in actual clinical settings is needed. A. ramosus could potentially be a future antimicrobial agent against Acinetobacter baumannii infections. Further research is needed to establish the actual conditions of such application. Also, the reason behind the lack of observation for the predation of A. ramosus against K. pneumoniae and P. aeruginosa remains to be established. Overall, our study demonstrates that predatory bacterias Bdellovibrio bacteriovorus and Agromyces ramosus species could be one alternative to the antibiotics which are increasingly becoming useless in the war against multidrug resistant bacteria and this study indicates that this option deserves further investigations

Sai Guo is a PhD student in soil microorganism and bio-organic fertilizer research innovation team of Nanjing Agriculture University, China. His supervisors are professor Qirong Shen and associate professor Rong Li. Dr Sai Guo has her expertise in soil microorganism ecology and interaction mechanism between microorganism and plants. He specializes in high-throughput data analysis of soil bacteria, fungi and protozoa. He and His team have been committed to improving the quality of Chinese soil arable land and rationally using bio-organic fertilizers to regulate the activity of soil microflora in the rhizosphere

Soil protists, a critical component of soil microbiome and useful bio-indicators for soil quality, are constantly overlooked in field soils, thus investigation of this important group will bring unpredictable contribution to agriculture industry. In this study, long-term field experiment with cucumber monoculture was adopted to disentangle the influence of Trichoderma enriched bio-fertilizers and organic fertilizers on soil protist community, compared with chemical fertilizers without extra microbial inoculation through the Illumina HiSeq sequencing. Results showed application of chemical fertilizers revealed lower protist richness and diversity than organic fertilizers and biofertilizers in both bulk and rhizosphere soils. Bray-Curtis distance principal coordinate analysis revealed significant differences in bulk and rhizosphere soil protist community structures, and the structures of chemical fertilizers were obviously separated from organic fertilizers and bio-fertilizers in both bulk and rhizosphere soils. Moreover, in bulk soils, organic fertilizers had the higher relative abundances of Alveolata, Amoebozoa, Archaeplastida and Opisthokonta than chemical fertilizers and bio-fertilizers. Compared to rhizosphere soils, Alveolata and Amoebozoa increased in bulk soil from chemical fertilizers, while the later one decreased in organic fertilizers. In addition, top 50 protist OTUs were assigned into functional groups, and OTU_144 (Acanthamoeba arachisporum) increased and OTU_54 (Hypotrichia: belonging to Omnivores) decreased from bulk to rhizosphere soils in all treatments. In sum, we concluded that field soil persistent management such as organic fertilizer and bio-fertilizer application could greatly alter bulk soil protist community and plant could further select protist community in the rhizosphere, which might be related with plant health and plant growth in the agriculture system.

Maria Pavlova is a senior assistant at the National Reference Laboratory of Enteropathogenic Diseases. She has experience in research, teaching and development of rapid molecular methods for diagnosis of campylobacteriosis and sallmonelosis direct from stool. Optimization and application of genetic methods for typing and detection of food- and water-borne epidemics. She has been a National Coordinator of Salmonellosis and Campylobacteriosis for the European Centre for Disease Prevention and Control. She is a research project leader and participant in two more

Introduction: The most frequent food and waterborne diseases affecting huge amount of people annually is due to Salmonella enterica subsp. enterica according to data from ECDC, CDC and WHO. In the summer of 2018 salmonella outbreak was recorded on the territory of the city of Ruse, affecting only medical surgical staff of two different healthcare establishments. The etiological agent of this outbreak is an exotic serotype for Bulgaria - S. London. The confirmation of epidemiological relatedness of the Salmonella isolates was made in the National Reference Laboratory of Pathogenic Diseases of the NCIPD through phenotypic and genetic methods. Materials & Methods: The total number of the surveyed persons (medical surgical staff of the two healthcare establishments) is 144. From 18 (18/144) of the tested suspicious persons were isolated etiological agent S. London, 2 of them (2/18) have had an acute clinical form and 16 (16/18) were asymptomatic. The serotype of isolates is phenotype confirmed as O:E: l, v : 1,6 by anti-sallmonella serums (BUL-BIO, NCIPD, Bulgaria). X-bal I PFGE analysis has been carried out to provide genetic evidence of the epidemiological relatedness of the salmonella isolates. Results & Discussion: The profiles of the PFGE analysis have shown that S. London isolates are epidemically related. The epidemic link between the affected medical surgical staff of two different healthcare establishments is determined by common factors - the food delivered in the two medical establishments is prepared by the same company, some of the affected persons are working in both medical establishments. There are no reported cases of laboratory confirmed salmonellosis among hospitalized patients in the affected surgical departments.

Bon Fabienne is a Research Teacher at the University of Burgundy. Her work aims to improve knowledge on the pathophysiology of Candida albicans and more specifically on the interaction of this agent with enterocyte cells. The objective is to understand the cellular and molecular mechanisms involved in the passage of Candida albicans from commensalism to the pathogenic state

Formerly a commensal resident of the gut microbiota in healthy humans, Candida albicans is an opportunistic pathogen causing infections ranging from superficial to the more life-threatening disseminated infections, especially in the ever-growing population of vulnerable patients in hospital settings. Because overwhelming evidences suggest that the gastrointestinal tract is the main source of disseminated C. albicans infections, deciphering the cellular and molecular mechanisms of the interaction of C. albicans with enterocytes is necessary to better understand the switch from commensalism to pathogenicity of this fungus and to improve the management of disseminated candidiasis. Different routes of the gut barrier translocation by C. albicans have been reported, including transcytosis through epithelial cells and the paracellular route where the fungus can cross the gut barrier without damaging the tissue barrier. The intestinal epithelium indeed consists in a monolayer of enterocytes where adjacent cells are jointed each other by cell junctions. Among those, tight junctions ensure integrity and impermeability of the intestinal barrier, limiting invasion of the gut barrier by Candida albicans. We propose to investigate the modulation of tight junction integrity during Candida albicans interaction with enterocytes and specify the mechanisms allowing the fungus to cross the gut barrier through the paracellular route.

Maka Muradashvili has been studying plant bacterial disease with different aspect. She is a Young Scientist and the subject of her PhD project was to study distribution and biodiversity of strains of Ralstonia solanacearum (causing plant bacterial wilt) in Georgia and to investigate control mechanisms. By the framework of a joint project with the NCDC, Georgia she has deposited in the GenBank the whole genome sequence based analysis of eight (GEO_6, GEO_55, GEO_57, GEO_81, GEO_96, GEO_99, GEO_230 and GEO_304) strains of R. solanacearum, which isolated in Georgia. The genome sequence data are a valuable resource for the evolutionary, epidemiological studies of this phytopathogen. Now her interesting areal is to researching plant antimicrobial extract as a biological control. She has been studying R. solanacearum specific bacteriophages collaborated with the Bacteriophage Institute, Georgia. They are actively involved in the research, which is still underway

Nowadays, the fight against plant diseases is a global problem, even though there are different approaches involving physical, chemical, biological methods have been elaborated and used. At the same time, the number of resistant microorganisms significantly increases. Plant extracts and phytochemicals with known antimicrobial properties can be of great significance in therapeutic treatments. In this regard, one of the potent members of the Asteraceae family is Stevia rebaudiana, which is popular in the world not only as a low-calorie, medicinal, natural sweetener, also known for its antioxidant activity of leaf extract. Stevia was introduced in Georgia in the 80s of XX century. In the implemented research was studied antimicrobial activity of Stevia plant extract against plant phytopathogens, which stored in the culture collection of the BSU, Institute of Phytopatology and Biodiversity, Georgia. The survey was conducted by the Ralstonia solanacearum, Pseudomonas syringae pv. Actinidiae and Erwinia Amylovora strains and fungal pathogen. To action of the extracts of Stevia leaves on the growth of bacterial strains was showed that the highest inhibitory activity (measured by zone of inhibition) was the ethyl alcohol and Chloroform. The largest zones of inhibition were observed for ethyl alcohol extract against Pseudomonas syringae pv. Actinidiae, KW1 strains, (20 mm) and Ralstonia solanacearum KhT88 and KhPe90 strains (18 mm).The results of the tests conducted on fungal pathogens also showed high activity chloroform and absolute ethanol extracts. Observations on 96 h cultures have shown that sensitive fungal pathogens are characterized by a decrease in growth compared to the control, spores and mycelium characterized deformation. Especially sensitive was Fusarium moniliforma to chloroform extract. The presented results allow to continue research in this direction and try these extracts not only in vitro but in vivo enviroment condintion

Baisuo Zhao is from Graduate School of Chinese Academy of Agricultural Sciences, China

The obligately anaerobic haloalkaliphilic bacterium Alkalitalea saponilacus can use xylan as the sole carbon source and produce propionate as the main fermentation product. Using mixed carbon sources of 0.4% (w/v) sucrose and 0.1% (w/v) birch xylan, xylanase production from A. saponilacus was 3.2-fold greater than that of individual carbon sources of 0.5% (w/v) sucrose or 0.5% (w/v) birch xylan. The xylanse is halostable and exhibits optimal activity over a broad salt concentration (2–6% NaCl). Its activity increased approximately 1.16-fold by adding 0.2% (v/v) Tween 20. To understand the potential genetic mechanisms of xylan degradation and molecular adaptation to saline-alkali extremes, the complete genome sequence of A. saponilacus was performed with the pacBio singlemolecule real-time (SMRT) and Illumina Misseq platforms. The genome contained one chromosome with a total size of 4,775,573 bps, and a G+C genomic content of 39.27%. Ten genes relating to the pathway for complete xylan degradation were systematically identified. Furthermore, various genes were predicted to be involved in isosmotic cytoplasm via the “compatible-solutes strategy” and cytoplasmic pH homeostasis though the “influx of hydrogen ions”. The halostable xylanase from A. saponilacusand its genomic sequence information provide some insight for potential applications in industry under double extreme conditions.

Yan Yanchun is a Professor and Doctoral Supervisor; She was granted Special government allowances of the State Council; She works in the Graduate School of Chinese Academy of Agricultural Sciences. Over last 20 years, she has completed seven national and provincial projects, obtained seven national invention patents and provincial awards for scientific and technological achievements. She has published more than 100 papers in influential academic journals in her research field, including 51 SCI papers and 1 monograph. Her team has been focused on bioremediation of environmental pollution for 20 years. More than 60 strains of bacteria or fungi capable of degrading pesticides or environmental estrogen chemicals have been isolated.

Phthalic acid esters (PAEs) are widely used as plasticizers to improve the flexibility of plastic products. The PAEs are carcinogenic, estrogenic and extremely difficult to be degraded under natural condition. The isolated strains Gordonia alkanivorans YC-RL2 and Mycobacterium sp. YC-RL4 were capable of degrading PAEs effectively, including dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), dicyclohexyl phthalate (DCHP), and di-(2-ethylhexyl) phthalate (DEHP). The optimal temperature and pH for DEHP degrading was 30°C and 8.0. Based on metabolites detected by HPLC-MS, the degradation pathway of DEHP was deduced where the strains transformed DEHP into benzoic acid (BA) via phthalic acid (PA) and mono (2-ethylhexyl) phthalate (MEHP). The alpha/beta hydrolase (DphM1) responsible for PAEs hydrolysis was identified from genomic library of YC-RL4. This enzyme had identity of 30%-40% with the known PAEs esterase, such as EstSP1, EstS1, EstG, M673 PAEs hydrolase, DphB and M11 PAEs hydrolase. DphM1 could hydrolyze all of 13 kinds of PAEs tested, especially ones with bulky side chain. The optimal catalytic condition of DphM1 towards PAEs was 30°C and pH8.0. Kinetic analysis showed that DphM1 preferred to DMP, DEP and DCHP with high catalytic efficiency, and also could degrade recalcitrant substance DEHP, DOP and BBP, which was the vantage of DphM1 compared to counterparts reported. Based on the result of molecular docking, DphM1 and DMP could interact perfectly with binding energy of -69.125/mol. Some hydrophilic amino acid such as His148, Ser210, Asp209 and His336 might contribute to binding substrate or catalysis. A series of mutants will be constructed to evaluate the function of putative active residues

She has her expertise in isolation and analysis of biological entities from complex environmental samples. She developed a protocol in which a target virus could be isolated from complex environmental mixtures and analyzed

Giant virus (GVs) parasitizing environmental acanthamoebae have been isolated from various humidic habitates (1, 2). We analyzed Pithovirus lacustris (alias KC5/2) and Urceolovirus corneum (alias KlaHel) which was first isolated from a sweet water reservoir Koblenz, Germany, and the cornea of a keratitis patient respectively, for their prevalence and immunogenicity. The lack of visibility of distinct cytoplasmic structures such as ribosomes against the background of their boarded corked keg-like bacteria size appearance rendered them as being of archea-like. Despite containing a bacterial genome sized dsDNA genome, their reproduction depends on infection and propagation within an amoeba host. Although DNA isolated from various global samples were PCR positive, culturing of these giant viruses succeeded merely for one sample collected by us. However, we observed substantial seropositiveness for both viruses from healthy blood donor samples, indicating potential ubiquity of the GVs after human contact with them in their environmental habitats. Despite lack of a capacity to elicit any pathology in mice, GVs challenge, elicits seropositivity and activates macrophages indicating carriage of an innate immune stimulatory potential. Specific aspects of it have been observed by us.