Day :
- Plenary Session
Location: Edinburgh, Scotland
Session Introduction
Gemma Saucedo Pagès
Aigües de Barcelona, Spain
Title: Implementation of new concentration technologies for microbiological recoveries in a drinking water system from Aquavalens Project
Biography:
Abstract:
A new concentration method for microbiological analysis has been implemented in a routine sampling and event or emergency situation in a Drinking Water System (DWS), including Drinking Water Treatment Plant (DWTP) and Distribution Network (DN). This concentration method has been developed during European Aquavalens Project (an EU funded -7th Framework Programme- that aims to protect the health of Europeans by improving methods for the detection of pathogens in drinking water and water used in food preparation). This new protocol is based on a hemodialyzer membrane filter, able to concentrate high water volumes and recover three different kingdoms (viruses, bacteria and protozoa). Field samples were taken from DWTP and DN on a monthly period during one year. Parametres such as E. coli, Campylobacter spp, Legionella spp and Legionella pneumophila, Norovirus GI/GII, Hepatitis A virus, Giardia spp and Cryptosporidium were tested by using molecular methods (qPCR and FISH). Obtained results from these new methodologies were compared to standarized and/or validated methods, showing better sensibility and recoveries in most cases.
Data is used to calculate treatment efficiency at DWTP (logarithmic reduction of the microbiological load at different treatment steps), to assess DN quality water through Water Safety Plans (WPS) and improving Standard Operational Protocols (SOPs). It is also a less time- consuming (sampling and analyzing) method, saving money and human resources.
Acknowledgments: This study has been funded by the European Union through the project AQUAVALENS (EU grant number 311846 www.aquavalens.org)
Xiu-Ling Wang
Agricultural University of Hebei, China
Title: Isolation, Oxygen-tolerant Domestication of Anaerobic Soy Isoflavone Metabolizing Bacteria and the Application of the Microbial Metabolites of Soy Isoflavones
Biography:
Xiu-Ling Wang went to South Korea as a Chinese and Korean Government Exchange Program Scholarship Student and has completed her PhD at Seoul National University. She is the disciplined Leader of Microbiology in Agricultural University of Hebei. In 2015, she was awarded the National Prestigious and Moral Standard Teacher by the Chinese Ministry of Education. She has published more than 20 papers in reputed journals and has been serving as an Editorial Board Member of both Chinese and English journals. She has obtained twelve government issued patents (the first inventor) since 2011.
Abstract:
Epidemiologic as well as laboratory studies have shown that dietary soy isoflavones can exert multiple biological activities, including anticarcinogenic, antimutagenic and reduced risk of cardiovascular disease and potential protective role for osteoporosis and menopausal symptoms. Soy isoflavones are mainly composed of genistein and daidzein and undergo extensive metabolism in the intestinal tract prior to absorption. There has been growing interest in the microbial metabolites of soy isoflavones due to their wider or stronger bioactivities. To date, numerous particular bacteria responsible for isoflavone metabolism have been isolated and identified. However, all the reported isoflavone metabolizing bacteria are obligate anaerobes, which lack the ability to grow and convert isoflavones aerobically. After a long-term domestication, we obtained several oxygen-tolerant bacteria, which not only grew aerobically but also convert soy isoflavones to different metabolites in the presence of atmospheric oxygen. The obtained oxygen tolerant mutant strains differed from the original obligate anaerobic bacterial strains by various characteristics, including change in bacterial shape, in biochemical traits, etc. Enhanced production of the soy isoflavone metabolites by the oxygen-tolerant mutant strains was investigated. Moreover, our results demonstrated that equol, one metabolite of soy isoflavone daidzein, induced apoptosis in human hepatocellular carcinoma through the intrinsic pathway and the endoplasmic reticulum stress pathway. 5-hydroxy-equol, one metabolite of soy isoflavone genistein, significantly enhanced the lifespan and stress resistance of Caenorhabditis elegans.
Guzin Candan Gurukan- Middle East
Technical University, Turkey
Title: Diversity of wild non-Saccharomyces yeast population in must and wine during spontaneous wine fermentation
Biography:
Guzin Candan Gurakan (Gultekin) is a Professor at Food Engineering Department, Middle East Technical University since 2010. She has completed experimental part of her PhD in German Culture Collection, Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany. She held short-term post-doc positions at Kent University, UK, TNO, The Netherlands.
Abstract:
There is an increasing trend to isolate, identify and use non-Saccharomyces yeasts in winemaking due to the improvement in aroma quality and complexity of the final wine products. In this study, presence of industrially important selected non-Saccharomyces yeasts were analyzed in must and during spontaneous fermentation intervals using real-time PCR. In addition, non-Saccharomyces yeasts were isolated from must and spontaneous wine made from 4 different grape types of 4 different provinces, in Turkey and identified by DNA sequencing. The Ä°dentifications were completed by sequencing analysis of 5.8S-ITS region (5.8S gene and intergenic regions ITS1 and ITS2) using ITS1 and ITS 4 primers and D1/D2 domain (rDNA regions of 26S gene) using NL1 and NL4 primers. 11 NS species belonging to 6 genera were identified. Metschikowia pulcherrima, Metschikowia sinensis, Metschikowia chrysoperlae, Lachancea thermotolerans, Wickerhamomyces anomalus, Hanseniaspora uvarum, Hanseniaspora opuntiae, Hanseniaspora guilliermondii, Hanseniaspora meyeri, Rhodotorula mucilaginosa, Starmerella bacillaris were the isolates of identified non- Saccharomyces yeasts from spontaneous wines of 4 different grape types.
Abdelfateh Kerrouche
Edinburgh Napier University, UK
Title: Towards the Development of New Integrated Ultrasonic Filtration Systems for Monitoring Waterborne Pathogens
Biography:
Abdelfateh Kerrouche received his PhD degree in Measurement and Instrumentation from City University London in 2009. He is a Lecturer and Sensors/Systems Consultant at Edinburgh Napier University. He has published more than 15 papers in reputed journals and international conferences. His area of interest includes: Pathogen detection, Sample prepearation, Sensors and Aquaculture.
Abstract:
The detection of waterborne pathogens mainly relies on sample processing. This process concentrates large volume of water to very small volume that can be used under microscopes. The current approved concentration techniques are slow, very expensive, time-consuming and often results in low recovery rates of pathogens. This research explores the use of high frequency sonication which is attached to two different types of filters to improve the recovery rate and work towards automated filtration/elution systems. Both systems were designed and tested with latex beads and live Cryptosporidium parvum oocysts. Results show that the recovery rate has improved by 22% on average when the ultrasonic transducer was used for the beads tests. Also, both systems show higher recovery rates with the use of megasonic energy; about 18% for the tangential flow filtration system and about 12% for the dead-end filtration system.
Habibu U. Abdu
Federal College of Education, Nigeria
Title: Agro-allied chemicals, Environmental Xenobiotics and Insecticides Resistance in Anopheles gambiae in Nigeria
Biography:
Habibu U Abdu received his BSc and MSc in Medical Microbiology from Bayero University Kano in Nigeria and a PhD in Molecular Entomology from University of Abertay Dundee, Scotland, UK. His major areas of research interest are medical microbiology and biotechnology, molecular entomology (insecticides resistance mechanisms in principal malaria vector in northern nigeria) and general biology.
Abstract:
Mosquito breeding sites were grouped into two different study zones (A and B) on the basis of human related activities taking place in and around the breeding sites. An. gambiae larvae collected from ecologically contrasting breeding sites were reared to adults in the laboratory. Adults from the F1 progeny were assayed for resistance against 4% DDT, 0.75% permethrin and 0.1% bendiocarb using the WHO adult insecticide susceptibility bioassay protocol. During mosquito sampling a survey was carried out in each site with the aim of documenting the most widely used insecticide. The levels of the physicochemical environmental factors were measured from the anopheline breeding sites. Results shows that pyrethroids (cypermethrin, lambda-cyhalothrin and cyfluthrin) and organophosphates (dichlovos, dimethoate and chloropyrifos) were most commonly used for crop protection in the agricultural sites, organochlorine (endosulfan and fipronil) and carbamates (carbofuran and carbaryl) were also used to a lesser extent. On the other hand, interview in the residential sites revealed indoor residual sprays (IRS), Piya Piya sprays (Piya Piya sprays are formulations produced locally as insecticeds sprays and without government approval) and coils containing pyrethroid insecticides with cypermethrin, lambda-cyhalothrin and cyfluthrin as common active ingredients were mainly used for personal protection. The results of measurement of physicochemical parameters showed little variation in the levels of the physical environmental factors (pH and temperature) across the sampling sites in the two zones studied. However, the levels of nitrates, nitrites, phosphates, sulphates and carbon content were higher in sites located in zone A than those in zone B. Overall, zone A is significantly different from zone B (p=0.000). There was evidence of high insecticides resistance among the mosquitoes tested from all the sampling sites. However, mosquitoes from agricultural sites (zone A) recorded higher insecticide resistance when compared to those from residential sites (zone B). These high levels of resistance are probably related to extensive pesticide usage in the zone. This is further supported by higher levels of the environmental chemicals recorded in zone A compared to zone B. These observations could have a significant impact on the environmental management and insecticide based approach to malaria vector control in Nigeria.
Samyah Jastaniah
King Abdulaziz University, Saudi Arabia
Title: Isolation, molecular characterization and extracellular enzymatic activity of culturable halophilic bacteria from hypersaline natural habitats
Biography:
Samyah D Jastaniah is currently an Assistant Professor in Microbiology, Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. He got his PhD from KAU with the thesis entitled: “Biosynthesis, purification and characterization of L-asparaginase from actinomycetes, isolated from Kingdom Saudi Arabia” in 2009. He has published many papers in reviewed journals. He is the supervisor of many MSc students on complementary researches. His areas of research interests are industrial microbiology and biotechnology, taxonomy of bacteria and the role of actinomycetes in biotechnology and medicine.
Abstract:
Saline habitats like Dead Sea have unusual extreme environments, due to their extreme salinity. Most saline habitats originate from the evaporation of seawater and have nearly neutral to slightly alkaline pH (such as the Red Sea pH 8.3 and Arabian Gulf, pH 8.3). Ten halophilic bacterial strains (two gram-negative) belonging to the family of Halomonadaceae and (eight gram-positive) belonging to the family of Bacillaceae were isolated from the Red Sea, Arabian Gulf and Dead Sea by subjecting the isolates to a high salinity medium followed by identification using 16S rRNA gene sequencing. Four of isolates were designated on the basis of their tolerance to high salinity; SBR1 exhibited 97% homology to Halomonas aquamarina, SBR2 showed 97% homology to Sediminibacillus sp., (Red Sea), SBA9 exhibited 94% homology to Halobacillus sp., (Arabian Gulf) and SBD17 gave 98% homology to Halobacillus dabanensis (Dead Sea). The isolates were also characterized by their physiological parameters, SBR1 showed optimum growth at 30°C, pH 8.5 and 1.5 M NaCl, SBR2 at 30°C, pH 6.0 and 1 M NaCl. Optimum conditions for SBA9 were 35°C, pH 6.5 and 1 M NaCl and for SBD17, 37°C, pH 7.0 and 1 M NaCl.
Reda Hassan Amasha
King Abdulaziz University, Saudi Arabia
Title: Use of NMR to Determine Compatible Solutes in Halophilic Bacteria Isolated from Highly Saline Areas
Biography:
Reda Hassan Amasha received her PhD in Molecular Biology and Biotechnology at University of Sheffield, UK in 2012 and she has done Diploma of Professional Development in teaching and learning in Center for Teaching and Learning Development from King Abdul Aziz University, Saudi Arabia, 2016. Her areas of research interest are in the occurrence of extremophile microorganism in non-extreme environments; molecular microbial diversity in caves and desert varnish; halophilic microorganisms and their environments and accumulation of compatible solutes as a strategy for adapting to salinity stress by using nuclear magnetic resonance (NMR) spectroscopy. She has published several papers in reputed journals.
Abstract:
Ten halophilic bacteria (two gram-negative) belonging to the Halomonadaceae and (eight gram-positive), belonging to the Bacillaceae, were isolated from the Red Sea, Arabian Gulf and Dead Sea using a high salinity medium, followed by identification using 16S rRNA. Four of the isolates were designated on the basis of their tolerance to high salinity. The isolates respectively exhibited 97% homology to Halomonas aquamarina, 97% homology to Sediminibacillus sp., (Red Sea), 94% homology to Halobacillus sp., (Arabian Gulf) and 98% homology to Halobacillus dabanensis (Dead Sea). 1H-NMR spectroscopy was used to determine the osmolytes accumulated by H. aquamarina, Sediminibacillus sp., Halobacillus sp. and H. dabanensis grown in a saline nutrient medium at varying concentrations of NaCl and a range of organic sources. In the case of H. aquamarina, betaine and ectoine concentrations increased at high salinities. In contrast, betaine was found when casein and peptone were used as nutrient sources, while ectoine was produced in the presence of peptone. In the case of Sediminibacillus sp., betaine was the only osmolyte produced at high salinities, while betaine and ectoine was produced when peptone and casein were used. In Halobacillus sp., betaine was the only osmolyte produced at high salinities, whereas betaine and ectoine were produced in the presence of peptone and casein. Finally, in the case of H. dabanensis, only betaine accumulated at high salinities and in the presence of all organic nutrient sources.
- Speaker Session
Location: Edinburgh, Scotland
Session Introduction
Sophie Vazulka
University of Natural Resources and Life Sciences, Austria
Title: Production of Antibody Fragments with Plasmid-based and Genome Integrated T7 E. coli Expression Systems – Evaluation of Systems Performance in Microtiter Fed-batch like Cultivations
Biography:
Sophie Vazulka is currently a PhD candidate at the University of Natural Resources and Life Sciences, Vienna in the Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli at the Department of Biotechnology (fermentation technology group), Vienna.
Abstract:
Although E. coli is the most prominent bacterial production host for recombinant proteins, some proteins with high economic potential can still hardly be produced at remunerative levels. We selected four different fabs (fragment antigen binding) (BIBH1, BIWA4, CIMZIA and FabX) with identical constant domains representing such challenging proteins. Fab yield can be affected by miss-folding, aggregation or unbalanced expression, translation and translocation levels of sub-units making it still challenging to efficiently design expression systems and production processes. For translocation to the periplasm a post-translational (OmpA) and a co-translational (DsbA) leader sequence were used. E. coli BL21(DE3) and E. coli HMS174 (DE3) were transformed either via pET vectors or genome integration. The resulting 32 clones were cultivated under fed-batch like conditions in the BioLector. Cell growth was not affected by leader/fab combinations but the yield of correctly folded fab ranged from 0 to 12.5 mg/g CDM. Higher expression rates caused higher amounts of free light chain and K12 strain reached higher yields. Except of CIMZIA with DsbA leader, genome integrated versions showed higher fab yields, reduced levels of free light chain and basal expression than plasmid-based systems. Independent of the expression system used, highest yields were obtained with CIMZIA, followed by BIWA4, BIBH1 and FabX. Leader sequence cleavage efficiency for DsbA was significantly lower than for OmpA, both showed lowest with CIMZIA. Summarizing, we showed that the selected set of host/gene dosage/leader/fab combinations resulted in a broad range of variation in terms of fab yields and processing and will be studied in detail during bench-scale fermentations.
Biography:
P Branco completed her Master’s in Molecular Biology at the Institute of Health Egas Moniz, Monte da Caparica, Portugal and PhD in Food Engineering by the Instituto Superior de Agronomia (ISA), School of Agriculture, University of Lisbon, Portugal. She has published eight papers in peer-reviewed journals. She is a Researcher in the HERCULES Laboratory at Évora University since 2017, where she focused on the development of new RNA-FISH probes for identification and detection of microorganisms involved in biodeterioration of cultural heritage in the Microtech - Art project funded by FCT.
Abstract:
Bio deterioration of cultural heritage (CH) materials (e.g., paper, marble, lime, mortar, parchment, metal, glass) was neglected for a long time since it was previously believed that it was only due to chemical and physical processes. However, over the last decades, it has been proven that the action of microorganisms is a critical factor in the deterioration process. Biodeteriogenic microorganisms cause serious aesthetical and structural damages in CH materials of inestimable value. Some of them can synthesize carotenoid compounds causing pink staining such as Rhodotorula sp., this yeast has been associated with the deteriorative effects observed in the Évora Cathedral, Portugal. To distinguish Rhodotorula sp., from other microorganisms that produce the same type of alterations on CH materials, proper identification methods must be applied. RNA-fluorescence and in situ hybridization (RNA-FISH) has the potential to specifically identify the target microorganism of interest in complex microbial communities (it is based on hybridization of fluorescently-labeled oligonucleotide probes targeting to specific regions of the ribosomal RNA). Thus, the aim of this study was to design a novel genus specific RNA-FISH probe against Rhodotorula sp., and to evaluate its specificity and performance both in silico and experimentally. This will contribute for facilitating Rhodotorula sp., identification in degraded CH materials by RNA-FISH. A novel probe for Rhodotorula sp., (L-S-Rh160-a-A-19-ATTO 647N, Rh160-ATTO 647N) was designed using decipher program. Its specificity was analysed in silico by nucleotide BLAST and its performance in terms of characteristics of the probe e.g., GC content and temperature of hairpin structures by oligonucleotide properties calculator oligocalc and hybridization efficiency by math FISH program. The experimental performance and specificity were evaluated by constructing the fluorescence-signal-response/formamide concentration curve for the target and non-target yeast (Cryptococcus adeliensis) and by testing the probe against several other non-target CH biodeteriogenic microorganisms. To do this, a previously described RNA-FISH procedure was applied and the results were analysed by flow cytometry (FC) and by epifluorescence microscopy (EM). In silico analyses of Rh160-ATTO 647N indicated that this probe has a high coverage and specificity for the target genus (796 matches of the target organism in 1000 sequences and only one match for organisms from the same ecosystem of the target organism, Cryptococcus sp.,); fulfills the criteria for being an RNA-FISH probe e.g., 52.60% of GC content and temperature of hairpin structures below hybridization temperature and shows a high maximal theoretical hybridization efficiency with Rhodotorula sp., (99.92% with 0% of formamide). The experimental results were in agreement with these in silico analyses revealing that Rh160-ATTO 647 N probe has a high specificity and performance without formamide. Maximal strong and intense fluorescence signals were detected by FC for the target and absence of signal for all the non-target microorganisms tested. Therefore, this study contributes to an easy and fast identification of Rhodotorula sp., yeast involved in the CH bio deterioration process by RNA-FISH. This will be advantageous for CH safeguard.
Iram Fatima
Quaid-i-Azam University, Pakistan
Title: Antimicrobial potential of aerial parts of Dichanthium annulatum and Ochthocloa compressa
Biography:
Iram Fatima is doing her PhD from Quaid-e-Azam University, Islamabad. She is currently doing part of her PhD research work at Newcastle University, UK under HEC (Higher Education Commission of Pakistan) International Research Support Initiative Program (IRSIP). She has published five papers in reputed journals and has presented her work in various international conferences.
Abstract:
Bahawalpur region is situated in the southern Punjab province in Pakistan. Present study was done to evaluate the biological potential of the methanol and n-hexane extracts of two species of Bahawalpur i.e., Dichanthium annulatum and Ochthocloa compressa belonging to family Poaceae. Phytochemical studies revealed that various secondary metabolites were strongly present in the methanol extracts of both species. Maximum phenols (263.345±16.35 mg GAE/g) and flavonoids (28.147±0.32 mg QE/g) contents were detected in the methanol extracts of O. compressa species. Antioxidant potential of plants was determined via free radical scavenging assays and the antimicrobial assays were performed using agar disc diffusion method. Results revealed maximum antioxidant potential in both extracts of O. compressa species as compared to the D. annulatum. Similarly, in case of antimicrobial assay, maximum inhibition zones were formed by the extracts of O. compressa against most of the tested bacterial and fungal strains. However, S. typhi, W. anomalus, A. niger and F. oxysporum were revealed as the most resistant strains and L. monocytogenes, B. spizizenii and A. flavus were revealed as the most sensitive strains. Overall, extracts of O. compressa showed significant biological potential as compared to the D. annulatum plant extracts. Hence, its crude extract can be used in pharmaceutical industries to cure various infectious diseases caused by resistant microbes.
- Poster Presentations
Location: Edinburgh, Scotland
Session Introduction
Varintip Srinon
Mahidol University, Thailand
Title: Burkholderia pseudomallei BimC is required for actin-based motility, intracellular survival and virulence
Biography:
Varintip Srinon has completed her PhD in 2018 from Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. She is the young researcher at Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand. She has published more than 4 papers in reputed journals.
Abstract:
The intracellular pathogen Burkholderia pseudomallei, the etiological agent of melioidosis in human and various animals, is capable of survival and movement within the cytoplasm of host cells by a process known as actin-based motility. The bacterial factor BimA is required for actin-based motility through its direct interaction with actin and by mediating actin polymerization at a single pole of the bacterium to promote movement both within and between cells. However, little is known about the other bacterial proteins required for this process. Here, we have investigated the role of the bimC gene (bpss1491) which lies immediately upstream of the bimA gene (bpss1492) on the B. pseudomallei chromosome 2. We have constructed a B. pseudomallei bimC deletion mutant and demonstrate that it is defective in intracellular survival in HeLa cells. This defect in intracellular motility in HeLa cells correlates with ablation of plaque and multi-nucleated giant cell (MNGC) formation. These defects in intracellular survival and cell to cell spread are not due to the loss of expression and polar localization of the BimA protein inside infected cells; however they do correlate with an inability of the bacteria to recruit and polymerize actin. We also establish a role for BimC in virulence of B. pseudomallei using a Galleria mellonella larvae model of infection. Taken together, our findings indicate that B. pseudomallei BimC plays an important role in intracellular behavior and virulence of this emerging pathogen.
Thamer Tbeez
Al Mustansiriya University,Iraq
Title: Correlation between Tansforming Growth factor Beta With Habitual abortion in women infected with Cytomegalovirus
Biography:
Thamer Mutlag Jasim completed his graduation in Veterinary Medicine at the University of Mosul, 1979-1980. He completed his Master’s degree in Medical Microbiology at College of Medicine, University of Tikrit in 1996. He completed his PhD at the University College of Medicine in Medical Microbiology in 2001. He has published more than 40 papers in peer reviewed journals and book titled Microbial Food Poisoning. He is an Assistant Professor at the College of Pharmacy, Al Mustansiriya University, Department of Laboratory Clinical Science since 2002. He has his expertise in infectious disease, counter immuno electrophoresis test, antibacterial nanotechnology, resistant bacteria, biofilm bacteria, herbs as antibacterial and anticancer and parasitic infection especially Toxoplasma gondii
Abstract:
Recurrent pregnancy loss (RPL) is the most frustrating and challenging field in reproductive medicine because the aetiology is often unknown and there are few evidence based diagnoses and treatment. The cytomegalovirus (CMV) has a ubiquitous DNA herpes virus, as with other herpes viruses, it becomes latent after primary infection but can reactivate with renewed viral shedding. The aim of the present study is to estimate the role of transforming growth factor beta 1 (TGFB1) to CMV immunoglobulin. The study was done in Kamal Al- Smarrai hospital in Baghdad, Iraq, from the period of October 2016 to February 2017. This study was performed on 88 pregnant women attended, 24 with unsuccessful abortion (two or more abortion) and 27 had single abortion and compared with 37 women with normal pregnancy were control, no recurrent abortion). Serum levels of TGF, IgG, IgM and IgG avidity for anti - CMV virus were measured in the serum. They used ELISA reader and electrochemiluminescence for CMV IgG avidity. There were no significant differences between the studied groups in their age, family history of abortion. Serum anti- CMV IgG was significantly higher in RPL and single abortion group compared to IgG TGFB1 in the studied groups. There was no significant difference in the median of IgG and IgM among different groups. There was no significant difference among different groups in their IgG avidity. There is inverse weak correlation between IgM and anti CMV IgG with TGF B1 in control group. There was no correlation between IgG IgM and IgG avidity with TGF B in recurrent abortion group. The current study showed a high proportion of pregnant women with past CMV infection. The RPL, anti-CMV IgM and TGFB were correlated directly with RPL patient compared with healthy control.
Alysaa Decker
The Pennsylvania State University, USA
Title: Root microbiomes of tropical trees: fungal dynamics affecting seedling performance
Biography:
Alyssa L Decker is a third-year undergraduate student at the Pennslyvania State University. She is pursuing a major in Biology and a minor in Microbiology. Her research is under the direction of James H Marden and focuses on the integration of soil microbiology and ecology.
Abstract:
Interactions between fungi and plant roots affect community dynamics and diversity. Tropical trees in Panama have shown to be negatively affected by soil from conspecific trees, apparently because of shared susceptibility to pathogens. Here, we test the hypothesis that root microbiomes that establish in Virola surinamensis seedlings vary depending on relatedness to the soil inoculum source and affect seedling growth performance. In a greenhouse experiment, seedlings were grown in: mother, male, other female (conspecific) and heterospecific. We isolated and sequenced fungal RNA from the ITS2 region in roots of 158 V. surinamensis seedlings using QIIME, MOTHUR and other bioinformatics tools in a custom pipeline, we assigned fungal taxonomy and relative abundance values. The abundance of specific fungal taxa within the community including Rhizophagus clarus, Chaetothyriales and Sordariomycetes were significantly elevated in particular source inocula. Abundances of some possibly pathogenic fungi (e.g., Sympoventuriaceae, Fusarium sp.,) were positively associated with maternal soil and hence candidates for negative effects of growing near a parent tree. Multiple fungal species were identified as being transmitted in a species-specific and/or sex-specific fashion. Abundance of certain fungal taxa was positively associated with the microbiome diversity and seedling growth performance (as was microbiome diversity itself), but there were no clear negative associations between particular fungal taxa and seedling growth. This work in conjunction with haplotyping of resistance genes in the same seedlings and mother is revealing mechanisms underlying important components of the community ecology and dynamics of tropical trees.
Selva Manikandan Athi Narayanan
Edinburgh Napier University, UK
Title: Biodesulphurisation of benzothiophene and dibenziothiophene by rhodococci isolated from oil contaminated soil
Biography:
Selva M Athi Narayanan is a PhD student at the Edinburgh Napier University, UK. He has worked on applied microbiology research projects including bacterial biodesulphurisation, peptide antibiotic producing bacteria for several years and has co-authored two papers in reputed journals during the time.
Abstract:
Biodesulphurisation (BDS) is an emerging technology that utilizes microorganisms for the removal of sulphur from fossil fuels. In this research, two rhodococci that were previously isolated from oil contaminated soils in Russia were found to possess robust desulphurisation activity against benzothiophene (BT) and dibenzothiophene (DBT) respectively. One strain was able to convert DBT to hydroxybiphenyl (2-HBP) with DBTO and DBTO2 as intermediates, whereas the other strain was able to convert BT into benzofuran, indicating that BDS reaction followed the well-known 4S pathway of desulphurisation. The species identity of the BT desulphurising strain and DBT desulphurising strain was confirmed as R. opacus and R. erythropolis respectively by 16S rRNA and gyrB gene sequence analysis and by whole genome sequence based OrthoANIu values (>95%). The DBT desulphurisation genes (dsz) of the R. erythropolis strain occurred as cluster sharing high similarity with the dsz operon of R. erythropolis IGTS8. The putative genes encoding the BT desulphurisation activity of the R. opacus strain was identified using comparative genomics. When cultured directly in a biphasic growth medium containing 10% model oil (hexadecane) or diesel containing 300 ppm sulphur, the R. erythropolis culture formed into an emulsion by interacting with the oil making it unsuitable for direct industrial application despite its desulphurisation potential. Whereas, the R. opacus culture formed distinct oil, biomass and aqueous phases which enabled easy extraction of the desulphurised oil with 80 fold reduced sulphur level, as measured by inductively coupled plasma - optical emission spectrometry (ICP-OES), making it a desirable strain for commercial application.
Yuichiro Otsuka & Masaya Nakamura
Forestry and Forest Products Research Institute, Japan
Title: Large Scale Production of 2-pyrone-4,6-dicarboxylic acid as a platform chemical from Lignin
Biography:
Yuichiro Otsuka has completed his PhD from Tokyo University of Agriculture and Technology and Postdoctoral studies at Jikei University School of Medicine. He is the Senior Researcher of Forestry and Forest Products Research Institute.
Abstract:
Lignin is the most abundant aromatic biomass polymers in nature. Utilization of this abundant biomass for high value products however is limited and associated with lignin processing. One practical way to utilize lignin is to convert it using the chemical or enzymatic reactions to produce utilizable monomers such as vanillin for flavorings or even bio-based feedstocks for bioplastics and other useful chemicals. Typically, though lignin extraction processes produce a wide range of aromatic compounds ranging from vanillin to vanillic acid, syringaldehyde, syringic acid and other mono- or di- aromatic compounds. Although there is some value to these chemicals, the target compounds must be purified, often requiring complex separation procedures. A more efficient system to utilize waste lignin, while producing a purer stream of high value products would be desirable. The soil bacterium, Sphingobium sp., SYK-6, is able to degrade various low molecular weight compounds derived from lignin and to metabolize these low molecular weight compounds to produce 2-pyrone-4,6-dicarboxylic acid (PDC). We focused on this metabolic intermediate as a raw material for the potential production of novel bio-based polymers. We have established a technology to produce PDC as a platform chemical from lignin monomers using bacterial metabolic engineering technology. In this study, we have also established large scale process for producing PDC from vanillic acid by using metabolome technology. Finally, we have successfully produced PDC about 80 kg from vanillic acid by single batch fermentation.
Biography:
Abstract:
Canine brucellosis caused by Brucella canis is a worldwide distributed zoonosis. Infection often results in abortion, orchitis, epididymitis, and discospondylitis. The 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) is currently the gold standard diagnostic tool for B. canis. Although it has been a widely used test, it detects IgG and IgM antibodies and has low sensitivity and specificity. The antigen used in this diagnostic test commonly cross-reacts with other pathogens like Escherichia coli O157: H7, Francisella tularensis, Vibrio cholerae, Salmonella N group y Pseudomonas maltophilia, and its production require a level III biosafety laboratory. As a limiting factor, this test is not commercially available in our country. For this reason, it is necessary to seek additional antigen candidates for the diagnosis of canine brucellosis with a methodology of easy access for use in the veterinary clinic. Our group has demonstrated high levels of the GroEL protein in the serum of animals experimentally infected with B. canis, suggesting its role as a candidate protein for detection in diagnostic tests. For recombinant protein preparation, specific primers were designed to amplify the GroEL gene and later cloning into the pQE60 plasmid. Further protein expression requires the E. coli M15 strain that harbors the plasmid pREP4 (lacIq repressor protein). Protein semiquantification by Western Blot will compare recombinant GroEL with native protein. Purification by FPLC (fast protein liquid chromatography) and assessment as the capture antigen by indirect ELISA will be performed with serum from experimentally infected dogs.