Day 2 :
Keynote Forum
Jory Lange
Robins Cloud, USA
Keynote: Blockchain: Accelerating traceback investigations in food poisoning outbreaks
Time : 09:00-09:30
Biography:
Jory Lange became a lawyer to help make our communities safer. He helps families who have been harmed by large corporations. Through his cases, he works to deter corporations from harming other families in the future. After 14 years of practice, Jory has wide-ranging experience in litigating cases involving dangerous products. As coleadof Robins Cloud’s Food Safety team, he helps victims of food poisoning. After graduating with honors from the University of Chicago, he earned his law degree at the University of Texas School of Law. While at the University of Texas, he was awarded the Deans’ Achievement Award for achieving the highest grade in Constitutional Law.
Abstract:
Statement of the problem: International food supply chains have made tracing contaminated food products back to their source difficult and time-consuming. Once a contaminated food product has been identified, investigators must first retrace its journey backwards from the retailer through one or more distributors to its original manufacturer, farm, or ranch. Once investigators have discovered the original source, investigators must then reverse course and trace the product’s route forwards through the supply chain this time from farm to manufacturer, through distributors, and to all its ultimate retailers. Currently, traceback investigations can take weeks or even months. Traceback investigations are complicated because: (1) there is no single database for tracking food products through supply chains from their original source to their ultimate consumers; (2) not all supply chain information is digital and critical supply chain information exists only in paper documents; (3) paper documents must be physically gathered to reviewed by investigators; (4) our food supply chains are now international, with food products routinely shipped from farms in one country to markets in another country or continent; (5) often investigators live in the market country, not the country of the product’s origin; (6) there may be language or interpretation barriers; and (7) the food supply chain has grown so complex that some retailers have great difficulty in guaranteeing the provenance of the food products they sell.
Blockchain’s Application to Supply Chain Management and Food Safety: Blockchain technology creates a way for investigators to rapidly trace contaminated food products backwards from fork to farm and then forwards from farm to fork. Traceback investigations that currently take weeks to months could be completed in minutes. By accelerating traceback investigations, blockchain technology can lead to much more rapid product recalls, reduce the spread of outbreaks, and ultimately, to fewer consumers being injured by contaminated food products.
Blockchain Explained: A blockchain is simply a continuously growing list of records (called “blocks”). Once data is recorded in a block, the data cannot be changed without also changing all subsequent blocks in the chain. This means there is a permanent, unalterable record. The blocks exist on a distributed ledger in which the records are distributed through a vast network of computers spanning the globe. This permanent, unalterable record can be accessed from anywhere on the planet.
Industry Experiments with Blockchain in Traceback Investigations: In May of 2017, Walmart reported the results of its collaboration with IBM on blockchain technology. Using blockchain technology, Walmart traced Chinese pork and U.S. mangoes back to their original sources. This traceback process, in the past, would take two weeks. But with blockchain technology, the traceback took 2.2 seconds. Walmart, Nestle, and Unilever are currently collaborating with IBM to apply blockchain to the global food supply.
- Oral Session 1
Location: Goya
Session Introduction
Natalija Sverchkova
Belarus National Academy of Sciences, Belarus
Title: Contribution of applied microbiology into national economy of Belarus Republic
Time : 11:05-11:25
Biography:
Natalija Sverchkova
Abstract:
Significance of applied microbiology is determined by the 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 in the evaluation of microbial biodiversity, physiology, and biochemistry; genetic engineering of microorganisms producing bioactive substances; development of biotechnologies for an industry, agriculture, medicine, environmental protection. A crucial aspect of microbiological processes is the provision of superactive strains synthesizing bioactive substances. Both conventional selection methods and genetic engineering are used to attain this goal. The Center for Analytical and Genetic-Engineering Studies set up at the Institute is equipped with up-to-date instrumentation allowing to derive strains with defined properties engaged in an 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 the preservation of genomes of promising biotechnological objects; early diagnostics of pathologies affecting forest and agricultural cultures; bioremediation of natural and technogenic environments; evaluation of diverse materials bioresistance. Technologies and production generated at the Institute find the outlet in numeric branches of national economy, including agriculture – biopesticides, microbial fertilizers, probiotics, biodisinfectants, biopreservatives, enzyme preparations; medicine – pharmaceutical technologies, diagnostic kits; industry – food additives, organic acids, alcohols, and makes a solid contribution to its progress.
Linda K Medlin
Marine Biological Association, UK
Title: Advances in molecular tools for routine monitoring of toxic algae and pathogens in aquatic ecosystems
Time : 11:25-11:45
Biography:
Linda K Medlin obtained her PhD 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 two additional FP7 projects Aqua 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 longterm 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 five countries for the toxic algae and six 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.
John Alderete
Washington State University, USA
Title: A recombinant chimeric protein comprised of immunogenic epitopes of metabolic enzymes is serodiagnostic target for Trichomonas vaginalis sexually transmitted infections
Time : 11:45-12:05
Biography:
J F Alderete received his PhD from The University of Kansas in 1978. He did Postdoctoral research at The University of North Carolina at Chapel Hill. He has published 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 nonviral 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 our investigations of the T. vaginalishost 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. 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.
Mariateresa Volpicella
Bari University, Italy
Title: Defining the prokaryotic biodiversity of the salterns of Margherita di Savoia (Italy) by amplicon based next generation sequencing analysis
Time : 12:05-12:25
Biography:
Mariateresa Volpicella has completed her 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:
Background: 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.
Objective: Defining the microbiota composition of the salterns of MdS in ponds with increasing salt concentrations. Methodology: 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 & 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.
Mihai Nita-Lazar
National Research and Development Institute for Industrial Ecology-ECOIND, Romania
Title: Detection of waterborne pathogenic bacteria using immunofluorescence technique
Time : 12:25-12:45
Biography:
Mihai Nita-Lazar has a wide expertise in the fields of Microbiology, Molecular Biology and Biochemistry mainly in prestigious laboratories such as the Institute of Microbiology, ETH, Zurich, Switzerland, Department of Biochemistry and Cellular Biology of Stony Brook University, New York, SUA, 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 published more than 40 ISI papers, four 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., Vibrio 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 antibodyantigen interactions. Three pathogenic bacteria such as Pseudomonas aeruginosa, Escherichia coli and Legionella spp. were detectedby immunofluorescence 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 techniques proved to be a reliable one, time-effective and sensitive for diagnosis and prevention of drinking water quality and waterborne bacterial disease.
Catalina Stoica
National Research and Development Institute for Industrial Ecology-ECOIND, Romania
Title: New cellulosic economical enzyme-substrate based diet of pullets
Time : 12:45-13:05
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. Nowadays, 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 celluloses based nutrients (substrates) and 2 cellulase food supplements (enzymes). The results showed a 50% efficiency of cellulose degradation rate during 1h for a 1:1 enzyme-substrate ratio, especially for the BioZyme M6000 food supplement. Moreover, the nutrients and the food supplements were microbiologically tested for the presence of Escherichia coli and Salmonella spp. The results showed the absence of Salmonella spp., 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 nutrient supplemented with cellulase food supplement.
- Special Session
Location: Goya
Session Introduction
Marie Filteau
Laval University, Canada
Title: A systems biology approach to reveal the sweet secrets of maple syrup quality
Time : 13:50-14:20
Biography:
Marie Filteau has her expertise in Microbial Ecology, Systems Biology and Evolution. She is a Professor at the Department of Food Science at Laval University in Quebec City, Canada. She has fundamental interests in microbial ecosystems, in microbial interactions, their role in microbiome functions, and their impact on food quality. Her applied research is focused towards maple and meat products.
Abstract:
Maple syrup is a natural sweetener produced from the concentration of maple sap during the North-American spring season. Over the last decades, maple syrup production transitioned from artisanal to industrial, leading to annual productions of nearly ten million gallons in Canada. The increased volumes now justify the need to address quality variation in the product. A range of color grades, but also flavors and defects are encountered in maple syrup. We previously showed that maple sap microbial contamination and predominant populations are correlated with syrup quality. However, the causality underlying these correlations remains to be explored. Are microorganisms directly responsible for changes in quality, or are they only reflecting changes in maple sap chemical composition driven by the tree metabolism? To improve our understanding of maple sap composition variation, we applied a systems biology approach using a collection of barcoded Saccharomyces cerevisiae deletion strains as a biological reporter. By competing these genetically modified yeasts in various maple sap samples, we identified metabolic pathways linked to nutrient utilization that vary over the collecting season, one of which is associated with a major flavor defect. We also identified allantoic acid as the principal nitrogen source in map sap, which may play a major role in modulating microbial contamination. The next step is to identify how these molecules directly contribute to maple syrup quality and how maple sap microorganisms respond to variations in these limiting nutrient sources. Such knowledge may ultimately help devise microbial-based strategies to improve maple syrup quality.
- Oral Session 2
Location: Goya
Session Introduction
Shaukat Iqbal Hashmi
National Aquaculture Group, Saudi Arabia
Title: Validation for total aflatoxins (aflatoxins B1, B2, G1 and G2) in shrimp & fish feed, shrimp hepatopancreaz and fish liver by ELISA
Time : 14:20-14:40
Biography:
Shaukat Iqbal Hashmi is working as head of Central Analytical Services at National Aquaculture Group, Saudi Arabia. He has completed his M.Sc. from University of Agriculture Faisalabad, Pakistan in the year of 2002. He has more than 13 years of experience in ISO 9001,ISO 14001, ISO 22000, ISO 17025 & HACCP, Gloab GAP, BAP certified organizations.
Abstract:
Aflatoxins are toxic substances and are fungal metabolites (Mycotoxins) produced by certain molds of the genus Aspergillus growing on several raw food commodities. Aflatoxins are highly toxic compounds and can cause both acute and chronic toxicity in humans and many other animals. Aflatoxins consist of about 20 similar compounds but only four are naturally found in foods. These are aflatoxins B1, B2, G1 and G2. Aflatoxin B1 is the most commonly found in food and the most toxic e.g. when lactating cattle and other animals ingest aflatoxins in contaminated feed, toxic metabolites can be formed and may be present in milk. These metabolites, aflatoxin M1 and M2, are potentially important contaminants in dairy products. US food safety regulations include a limit of 20 μg/kg for total aflatoxins (B1, B2, G1 and G2) in all foods except milk and a limit of 0.5 μg /kg for M1 in milk. Higher limits apply in animal feeds. If fish or shrimp are fed by contaminated feed, ingestion of such feed can cause toxic metabolites contamination and can be found in liver of the fish and hepatopancreaz of the shrimp. It is very important to assess the quality of these products by monitoring the aflatoxins residue in fish liver and shrimp hepatopancreaz. There are a variety of methods available for aflatoxins determination e.g. by ELISA or by HPLC but the real challenge is to extract the aflatoxins from liver or hepatopancreaz of fish & shrimp. For extraction of aflatoxins from meat matrix, liver, kidneys etc., 70 % methanol is recommended. Helica Total Aflatoxins protocol for ELISA was followed for analysis and validation. Total aflatoxin Assay is a solid phase direct competitive enzyme immunoassay. An aflatoxin specific antibody optimized to cross react with all four subtypes of aflatoxin was coated to a polystyrene micro well of the ELISA plate. Aflatoxins were extracted from sample with 70% methanol. The extracted sample and HRP-conjugated aflatoxin B1 were mixed and added to the antibody-coated micro well. Aflatoxin from the extracted sample and HRP-conjugated aflatoxin B1 compete to bind with the antibody coated to the micro well. Micro well contents were decanted, and non-specific reactants were removed by washing. An enzyme substrate (TMB) was added and color (blue) was developed. The intensity of the color was directly proportional to the amount of bound conjugate and inversely proportional to the concentration of aflatoxin in the sample or standard. Therefore, as the concentration of aflatoxin in the sample or standard increased, the intensity of the blue color decreased. An acidic stop solution was added which changed the chromogen color from blue to yellow. The micro wells were measured optically by a microplate reader (ELISA reader) with an absorbance filter of 450nm (OD450). The optical densities of the samples were compared to the OD's of the kit standards and an interpretative result was determined.
Alejandro Garrido-Maestu
International Iberian Nanotechnology Laboratory, Portugal
Title: Isothermal DNA amplification for Salmonella spp. detection and characterization
Time : 14:40-15:00
Biography:
Alejandro Garrido Maestu is a Research Fellow working at International Iberian Nanotechnology Laboratory, with the expertise in Microbiology and Molecular Biology. At present he is working in a project based on a novel DNA amplification technique (loop-mediated isothermal amplification) for foodborne pathogens detection, combining with microfluidics and gold nanoparticles. This project is expected to have a great impact in the food industry as it will allow fast and accurate detection of the main foodborne pathogens. He has participated in several research projects involving bacterial agents detection, back in Spain and in the University of Florida where he started his Postdoctoral training. He has authored 10 peer-review papers with an h-index of 8, along with 1 book, all focused on food microbiology and bacterial pathogens.
Abstract:
Statement of the Problem: The genus Salmonella continues to be a major health issue. In Europe, more than 90000 of salmonellosis cases were confirmed in 2015 (1.9% increase respect to 2014), being serovar Enteritidis and Typhimurium the most prevalent ones, representing 45.7% and 15.8%. Conventional culture methods are lengthy and time-consuming, for this reason, the development of new methods that allow early detection of these pathogens is of high interest. Molecular methods have the ability of overcoming the previous limitations, being DNA amplification techniques the most attractive approach. In this sense, in recent years isothermal DNA amplification techniques have gained a lot of interest over other more stablished techniques, such as PCR/ qPCR, as providing several added-on advantages (no need of expensive equipment, many alternatives for product detection, etc.) One of these techniques with increase interest is loop-mediated isothermal amplification (LAMP).
Methodology: A selection of the most appropriate genetic targets was done, based on previously published studies, being selected invA for Salmonella spp. detection, STM4497 and safA for Typhimurium and Enteritidis serotyping respectively. Specificity and sensitivity assays were accomplished to ensure correct performance, and finally were implemented as the detection strategy after a simple pre-enrichment and DNA extraction. These new methods were compared against qPCR.
Findings: The newly designed assays performed excellent in terms of specificity, but presented lower sensitivity than qPCR. This problem was overcome with a pre-enrichment step. The evaluation of the performance of the assays in spiked food samples (turkey, chicken and egg) indicated that they may be implemented in routine food-testing. Conclusion & Significance: The developed methodology is suitable for routine food analysis, and may be used, either in a sequential mode or for directly assessing the presence of a specific serovar. LAMP represents a valid alternative to conventional DNA amplification techniques, and will allow costs reduction.
Rui Feng Mao
Guangxi University, China
Title: Analysis of microbial harm in sugar cleaning area of sugar mill
Time : 15:00-15:20
Biography:
Rui-feng Mao, was born in 1963, Guilin. In 1983, he has graduated from the Department of biology, Sichuan University, and received a bachelor of science degree in microbiology. In 2011, he has completed his doctoral degree from the school of chemistry and chemical engineering of Guangxi University and received his Ph.D. in engineering. He is currently an associate professor of food science at the school of light industry and food engineering of Guangxi University. He has long been engaged in the teaching and research work of microbiology and biochemistry. The main research areas include microbial isolation, screening, utilization of renewable resources.
Abstract:
In Guangxi of China, GMP (Good Manufacturing Practice) have been introduced in sugar mills since 2005 to comply with the requirements of the government administration, the clean zone was defined, and physical isolation was enforced as a procedure in sugar production. However, the rate of microbial detection in sugars did not reduce as expected. In this paper, the microbiological investigation and microbiological harm analysis of sugar clean zone in three sugar mills in Guangxi were reported. The methodology of investigating microbial species isolated from sugar cane clean area and its surrounding environment in sugarcane mill, as well as the distribution pattern of environmental microorganisms combined with air flow analysis and microbiological analysis were introduced. The research results provide the basis for control theory and control technology in the analysis of microbial harm in sugarcane mill.
Maria Turtoi
Dunarea de Jos University of Galati, Romania
Title: Pulsed light inactivation of wheat grain native microbiota and effect on germination capacity
Time : 15:20-15:40
Biography:
Maria Turtoi is a staff member of Dunarea de Jos University of Galati, Romania. With a solid background training in food science and engineering, she teaches courses such as food unit operations, food packaging, nonconventional technologies and food safety in minimal processing. Her research activity closely follows the teaching activity. However, she has expertise in emerging technologies such as pulsed light and ultraviolet light treatments used to decontaminate the surface of food and food contact materials with the purpose of extending the shelf life of treated food, and enhancing the safety of minimally processed food.
Abstract:
Statement of the problem: Wheat grains are naturally contaminated with bacteria, yeasts and moulds while in the field and during storage, which represent the major cause of grain spoilage. In particular, fungi produce losses related to heating, biochemical changes, production of mycotoxins, decrease of germination capacity, and loss of dry matter. Growth of microorganisms can be prevented by drying of grains to reduce the moisture content and using of chemical preservatives. However, drying methods are expensive and chemical preservatives affect the germination capacity of wheat grains. Pulsed light (PL) is an emerging non-thermal technology able to reduce the microbial population on the surface of food and food contact materials.
Methodology: Wheat grains have been exposed to PL at different fluencies (2.23–30.05 J/cm2) and different number of pulses (10–60) in an experimental set-up able to rotate the wheat grains and expose the entire surface to PL. Survival populations of fungi and bacteria have been counted after each PL treatment and compared with fungi and bacteria load of control sample. Germination tests have been performed for PL treated samples and control. Subsequently, wheat grains used to determine the germination capacity have been dried, ground and subjected to analyse antioxidant capacity and enzymatic activity.
Findings: PL exposure of wheat grains resulted in reductions of up to 2.3 log CFU/g and 1.2 log CFU/g for fungi and bacteria respectively. Germination capacity of wheat grains has not been affected by PL treatment, but germination rate has been improved due to reduction of microbial load and enhanced of enzymatic activity.
Conclusion & Significance: Results confirm the positive effect of PL treatment on wheat grain decontamination and enhancement of germination rate which extend the shelf life of grains and use in food industry.
- Workshop
Location: Goya
Session Introduction
Amparo Gamero
Institute of Food Science and Technology– Spanish Research Council, Spain
Title: Non-conventional yeasts in bakery
Time : 15:55-16:45
Biography:
Amparo Gamero holds a PhD in Food Science and Technology. She works at the Institute of Food Science and Technology of the Spanish Research Council. Her research topic deals with the study of the role of Saccharomyces and non-conventional yeasts in aroma production during food fermentations. She has several SCI publications and has active participation in international conferences.
Abstract:
Saccharomyces cerevisiae is the most common species used in bakery due to its rapid sugar consumption and CO2 production, the most important attributes required to leaven the dough. These attributes have been shown not to be unique to Saccharomyces cerevisiae, but also found in several non-conventional yeast species. Only a reduced number of these non-conventional yeast species presenting potential to be used in bakery have been described. The rest of them remain poorly studied, being a vast untapped potential for the use as leavening agents and flavor producers. In this research work, the potential of several non-conventional yeasts to be used in bakery was assessed employing dough-like conditions in the microbread platform. This platform allows fast screenings of different recipes. In this case, the capabilities of bread leavening and aroma formation of different non-conventional yeast species were tested. The results showed that bread leavened with the non-conventional yeasts Kazachstania gamospora and Wickerhamomyces subpelliculosus had better overall results compared to control baker’s yeast, especially regarding aroma profiles. In this way, this study points out Kazachstania gamospora and Wickerhamomyces subpelliculosus as interesting alternative baker’s yeasts.
- Workshop
Location: Goya
Session Introduction
Santiago Benito
Polytechnic University of Madrid, Spain
Title: Non-Saccharomyces yeasts Lachancea thermotolerans and Schizosaccharomyces pombe applications in wine food microbiology. Influence in wine quality
Time : 09:30-10:10
Biography:
Santiago Benito has his expertise in Wine Microbiology. He is Vice Director of the Polytechnic University of Madrid Chemistry and Food Technology Department and Director of the Polytechnic University of Madrid Winery Center. He has written more than 40 indexed publications regarding the wine microbiology topic.
Abstract:
The classical way to make red wine is based on the use of Saccharomyces cerevisiae yeasts during alcoholic fermentation and Oenococcus oeni bacteria during malolactic fermentation. This traditional winemaking methodology produces commercial stable red wines from a microbiological point of view. This work explains the use of a new red winemaking biotechnology that uses the combination of Lachancea thermotolerans and Schizosaccharomyces pombe yeasts as an alternative to the conventional alcoholic and malolactic fermentations. Schizosaccharomyces pombe consumes malic acid while Lachancea thermotolerans produces lactic acid to avoid an unnecessary deacidification in low acidic musts from warm viticulture areas such as the south of Spain. This methodology also reduces some malolactic fermentation hazards for human health such as biogenic amines and ethyl carbamate while it improves the colour and aroma profile.
- Oral Session 1
Location: Goya
Session Introduction
Ala Mohan
The New Zealand Institute for Plant & Food Research Limited, New Zealand
Title: Comparative analysis of New Zealand Listeria monocytogenes isolates using PFGE, MLST, MVLST and whole genome sequence analysis
Time : 10:25-10:45
Biography:
Ala Mohan is working as a research scientist at The New Zealand Institute for Plant & Food Research Limited, New Zealand.
Abstract:
Listeria monocytogenes is a human food-borne pathogen that causes serious illnesses including encephalitis, meningitis, septicemia, abortion and still birth but the disease is also manifested as gastrointestinal illness. Listeria monocytogenes is widespread in the environment, has been isolated from various ready-to-eat food products and most of the infections have been found to be foodborne. Molecular typing tools, population genetics and evolutionary studies have played major roles in understanding the transmission, different subtypes, the virulence and evolution of Listeria monocytogenes in different niches. Here, we compare 123 Listeria monocytogenes isolates from New Zealand seafood, horticultural and clinical sources with publicly available international isolates to better understand the characteristics of New Zealand isolates and their relationship with the international isolates. We conducted pulse field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), multi-virulence-locus sequence typing (MVLST) and whole genome sequence analysis (WGS) using single nucleotide polymorphism (SNPs) of NZ isolates and compared them with publicly available typings. PFGE analysis showed that the seafood isolates from NZ grouped separately from the global ones. The minimum spanning tree conducted using seven housekeeping MLST alleles revealed that the NZ isolates formed unique complexes or sequence types (ST) in comparison with the international isolates although some of the seafood isolates grouped closely with international human pathogenic lineage I group strains. Likewise, WGS analysis revealed that most of the seafood isolates clustered with lineage II of the international isolates and some with lineage I while some of the horticultural isolates were lineage III/IV. MVLST also showed unique STs from NZ that were not reported from other countries. Results from the MVLST analysis raises a speculation that the virulence genes may have evolved differently to adapt to NZ conditions and may involve a niche adaptation mechanism.
Phillipe P Minnaar
Agricultural Research Council, South Africa
Title: Schizosaccharomyces pombe and Saccharomyces cerevisiae yeasts in sequential fermentations: Effect on phenolic acids of fermented kei-apple (Dovyalis caffra L.) juice
Time : 10:45-11:05
Biography:
Phillip P Minnaar is an expert in the study of polyphenols in fruit, fermented fruit and alcoholic beverages with a focus on the prevention of the depletion of polyphenols during fermentation. He also studies the impact of agricultural practices on the polyphenols on fruit crop. He is an Applications Chemist specializing in Chromatography.
Abstract:
Kei-apple (Dovyalis caffra) is an evergreen tree indigenous to Southern Africa. The fruit contains high concentrations of L-malic acid, ascorbic acid, and phenolic acids. Kei-apple juice was sequentially inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae yeasts. A reference fermentation using only S. cerevisiae was included. The fermentation was monitored by recording mass loss. At the end of fermentation, twelve untrained judges conducted free choice aroma profiling on the fruit wines. The kei-apple juice and wines were analyzed for total titratable acidity, total soluble solids, pH, alcohol, L-malic acid, and phenolic acids. Total titratable acidity was ca. 70% lower in Kei-apple wines produced with Sc. pombe+S. cerevisiae than in kei-apple juice. Kei-apple wines produced with Sc. pombe+S. cerevisiae showed substantially lower concentrations of L-malic acid than kei-apple wines produced with S. cerevisiae only. Wines produced with S. cerevisiae only proved higher in phenolic acid concentrations than wines produced with Sc. pombe+S. cerevisiae. Chlorogenic acid was the most abundant phenolic acid measured in the kei-apple wines, followed by protocatechuic acid. Judges described the kei-apple wines produced with Sc. pombe+S. cerevisiae as having noticeable off-odours, while wines produced with S. cerevisiae were described as fresh and fruity. Kei-apple wines (Sc. pombe+S. cerevisiae and S. cerevisiae) were of comparable vegetative and organic character. Saccharomyces cerevisiae produced kei-apple wine with increased caffeic, chlorogenic, protocatechuic, and sinapic acids, whereas Sc. pombe+S. cerevisiae produced kei-applewines with increased ferulic, and p-coumaric acids and low L-malic acid.
Michela Favretti
Istituto Zooprofilattico Sperimentale delle Venezie, Italy
Title: Efficacy of domestic cooking of experimentally infected bivalve shellfish: A case study
Time : 11:05-11:25
Biography:
Michela Favretti graduated in Veterinary Medicine in 2001 from the University of Padua. She is specialized in “farming, hygiene, disease control of aquatic species and related products” (2005, University of Padua). She is currently a Veterinary Manager and Quality Manager at the IZSVe laboratory in San Donà di Piave (Venice). Her fields of interest are food safety, microbiological analysis of foodstuffs and the simplified own-check systems to control hygiene. Her research interests include food safety, the promotion of local traditional products, simplification of the own-check system to control hygiene in small businesses (bars, restaurants, butchers, fishmongers, etc.) and small dairies. She is also a Trainer for food industry professionals and trade associations. She drafts guides of good manufacturing practices for small local producers about vegetables, fresh and processed meats, honey, milk and dairy products, processing in alpine hut.
Abstract:
Bivalve shellfish filter large volumes of water and in addition to retaining food particles they can accumulate chemical and biological contaminants from their surrounding environment. The consumption of raw or lightly cooked molluscs represents a health risk due to the possible presence of Escherichia coli, Vibrio spp., norovirus, hepatitis A virus, Salmonella and other pathogens, which could be present in the marine environment as result of fecal contamination. The present study was aimed to validate the instructions for domestic cooking to be labeled on clams (Tapes semidecussatus) and mussels (Mytilus edulis) in order to reduce or eliminate microorganisms artificially inoculated in the bivalve shellfish. A mix of Salmonella Senftenberg, Escherichia coli and Vibrio parahaemolyticus was dissolved in a bin containing 70 liters of sea water collected from the harvesting area of clams and mussels, and kept at 15°C; the bivalves were dipped in the contaminated water for 90 minutes. Inoculated microorganisms were counted in sea water before the immersion of molluscs, and in bivalve samples at the end of dipping time to verify the contamination levels. The absence of the same bacteria in non-contaminated bivalves (negative controls) was also confirmed. Contaminated clams and mussels were cooked in a pan with cover for 5 minutes once a temperature of 90°C was reached as suggested by the producer. At the end of the set time all the clams and mussels had opened valves; immediately they were shelled and pooled to be tested for the surviving microorganisms. The experiment was repeated in two sessions and two different batches of molluscs were used. The findings confirmed the total inactivation of artificially inoculated microorganisms in both bivalve species if cooked at 90°C for 5 minutes; so this time-temperature combination represents correct cooking instructions that can be labeled and guarantees a safe consumption of bivalve shellfish at domestic level.
Margit Olle
Estonian Crop Research Institute, Estonia
Title: Effective microorganisms improve the growth and quality of vegetables and soybeans
Time : 11:25-11:45
Biography:
Margit Olle finished Estonian Agricultural University in 1993 Cum Laude passing all exams excellently. She pursued her Doctoral studies in 1995 – 1999 in Norway at the Agricultural University of Norway and defended her Doctor Scientiarum degree. Since 2008, she has worked as a Senior Researcher at the Estonian Crop Research Institute (former Jogeva Plant Breeding Institute). She has published numerous review articles in addition to research articles in her field. She has published four books, out of them three in English and one in Estonian. She has been invited speaker in South Korea, Poland and Greece. Her biography is present in Who is Who in the World 2015 and 2016 (32, 33 Edition). Her CV is in the 2000 outstanding intellectuals of the 21st Century, 9th Edition, 2015.
Abstract:
Background: EM (effective microorganisms) is a combination of beneficial naturally occurring microorganisms, is a liquid concentrate. EM is produced from cultivations of over 80 varieties of microorganisms. Beneficial effects of EM are: EM promotes germination, flowering, fruiting, and ripening in plants and it improves the physical, chemical, and biological environments of the soil and suppresses soil-borne pathogens and pests.
Open land experiments results: The yield of beet root and swede increased with EM treatment, while yield of white and Chinese cabbage was not different from control. Beet roots and cabbages had less storage loss by EM treatment, while swede storage loss was not different from control. The content of calcium was higher in EM treated vegetables. The content of sugar and C-vitamin was higher in EM treated vegetables. The content of nitrates was higher in EM treated vegetables. Winter garlic yield was higher in EM treated vegetables and soil decomposing in fields using EM was much easier. The clover has much more roots and more Rhizobium nodules on roots in EM treated field.
Greenhouse experiments results: EM improve the quality of tomato transplants, as they remained more compact with a greater stem diameter (my discovery, not a patent unfortunately). In tomato transplant leaves the contents of nitrates, nitrogen, phosphorus, potassium, calcium and magnesium were higher in EM treatment. Cucumber, squash and pumpkin height was shorter and stem diameter greater by EM treatment and Onions developed more leaves by EM treatment.
Soybean experiments results: Soybean gave more yields by EM treatment and soybean germination was much better by EM treatment.
Zhu Lixia
NNSFC Foundation, China
Title: Diversity and transmission of Saccharomyces cerevisiae population in Xinjiang province, China
Time : 11:45-12:05
Biography:
Zhu Lixia has been engaged in research work of food microbiology and traditional fermented food for more than 10 years. She has her expertise in Xinjiang Msalais, a traditional wine spontaneously fermented boiled grape juice, including in wild yeast diversity and fermentation characteristics, aroma characters, chemical profile and their evolutions during Msalais process. She has more than 20 paper published in English and Chinese.
Abstract:
Statement of the Problem: Saccharomyces cerevisiae is the main driving force of traditional food fermentation; Both geographic differentiation and human activities drive the global evolution of S. cerevisiae, but its domestication origin is keeping unknown as an academic dispute. The Silk Road combined diverse geographical features and diversity civilization is most possible to be pregnant with unique genotypic - phenotypic features of indigenous S. cerevisiae population. Xinjiang in China is an indispensable part of Silk Road.
Methodology & Theoretical Orientation: S. cerevisiae regional characteristic was studied from Xinjiang, China, with the methods of SSR and multiple gene sequencing.
Findings: The diversity of S. cerevisiae associated with grape in Xinjiang was very rich beyond our thoughts; there were tree unique groups different from European wine strains and Chinese S. cerevisiae. The regional relationship of S. cerevisiae population in northern Xinjiang was closer than in southern Xinjiang, the migration direction of S. cerevisiae population was referenced from east to west and from north to south in Xinjiang and was more impacted on by human activity than climates, although both of them were important for the relational characters of indigenous S. cerevisiae populations.
Conclusion & Significance: The relationships of S. cerevisiae population in Xinjiang tested in the study would afford a firm scientific reference to identify the terrio grape or wine and to analyze the evolutionary characters of S. cerevisiae.
- Oral Session 2
Location: Goya
Session Introduction
Sanin Musovic
Danish Technological Institute, Life Science, Denmark
Title: The Trojan Horse: A new biotechnology for pesticide removal at drinking water sand filters
Time : 12:05-12:25
Biography:
Sanin Musovic is a Molecular Microbiologist, completed his PhD in 2010 and 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 a reviewer for five international journals, external/invited expert in EU commissions LIF Team, has over 30 publications in 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 a significant role in the removal of pollutants, where a sudden change in e.g. ammonium removal suggests a misbalance in a 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 the 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 the bacterial population. AOB was 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 copper appeared to be the product of increased catabolic abilities of the existing AOB cells.
Sergio Giannattasio
Institute of Biomembranes, Bioenergetics and Molecular Biotechnologies, Italy
Title: Identification of active compounds for the development of new anti-prostate cancer drugs: Comparative study in yeast and human cells
Time : 12:25-12:45
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 include mitochondria-to-nucleus cross-talk in cell stress response and the role in tumorigenesis of onco-suppressor 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 onco-suppressor 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.
Iris Yedidia
Plant Sciences – Agricultural Research Organization, Volcani Center, Israel
Title: Interkingdom signaling: Interference of plant derived small molecules with bacterial communication and virulence
Time : 12:45-13:05
Biography:
Iris Yedidia is the Head of the Ornamental Plants and Agricultural Biotechnology Department at the Agricultural Research Organization (ARO), Volcani center, Israel. Since 2004, She is 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 a diverse array of low molecular-mass compounds, with more than 8,000 phenolics. Phenolic compounds play a pivotal role in plant defense and fitness. Here, we provide an example of the 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 the interaction of the plant compounds with bacterial QS targets, drug discovery tools were occupied using the schrodinger® molecular docking suit. Pectobacterium central QS proteins ExpI/ExpR were used as targets for salicylic acid and carvacrol (plant phenolics). Finally using isothermal calorimetry (ITC), SA and CAR were directly 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 signaling shown here supports a rising field of research endeavoring interkingdom communication between plants and bacterial pathogens.
David Sartory
SWM Consulting Ltd, UK
Title: Evaluation of a rapid and simple MPN method for enumerating Legionella pneumophila from water
Time : 13:50-14:10
Biography:
David Sartory is an Independent Consultant, and 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 eight years in South Africa, where he obtained his MSc in Limnology. Prior to his current role, he was a 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 seven 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.
Sanin Musovic
Danish Technological Institute, Denmark
Title: Integration of natural microorganisms at drinking water sand filters with groundwater composition
Time : 14:10-14:30
Biography:
S Musovic is a Molecular Microbiologist, completed his PhD in 2010 and 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 a reviewer for five international journals, external/ invited expert in EU commissions LIF Team, has over 30 publications in 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 (MGEP) would ensure transfer and permanent establishment of introduced MGEP 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).
Rasih Felek
Akdeniz University Hospital Central Laboratory, Turkey
Title: Evaluation of Aspergillus immunochromatographic lateral flow device and galactomannan antigen test for the presence of invasive aspergillozis in patients with haematologic malignancies
Time : 14:30-14:50
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.
- Young Research Forum
Location: Goya
Session Introduction
Tugce Ulutasdemir
Sakarya University, Turkey
Title: Investigation of effect of edible coating containing Williopsis saturnus var. saturnus on aflatoxin production in peanuts
Time : 14:50-15:05
Biography:
Tugce Ulutasdemir graduated from ManisaCelal BayarUniversity, Department of Food Engineering in 2015. She started her graduate studies in Sakarya University Department of Food Engineering. She is currently working as a food engineer at BagdatlilarCerez. Work areas include food microbiology, edible films, biodegradable packaging, biocontrol applications.
Abstract:
During harvest, drying, and storage process of peanut (Arachis hypogaea L.), if its water activity exceeds the critique level fungus will grow and produce secondary metabolite called aflatoxin. Aflatoxin produced by Aspergillus flavus is mycotoxin which is carcinogenic, teratogenic and hepatoxic. Many scientific studies have shown that Williopsis saturnus var. saturnus yeast as a killer yeast has antagonistic antifungal effects. In this study, effect of edible coating incorporated with Williopsis. saturnus var. saturnus on the growth of A. flavus and aflatoxin production on peanuts. Whey protein concentrate (10%) and 2% glycerol were dissolved in water and heated for 30 min. After cooling, Williopsis. saturnus var. saturnus (10 logs CFU/mL) was added into the coating solution. The roasted peanuts were coated with coating solution with and without yeast, distilled water containing yeast (10 logs CFU/mL) or water as control by spraying. The peanuts were dried at 40 ºC in an oven. Following drying, the surface of peanuts was contaminated with A. flavus (4 logs CFU/g). The contaminated peanuts were dried, packed and stored for 90 days at 25 ºC. Every other week, in the peanut samples, the number of yeast and mold, the amount of aflatoxin, TBA (thiobarbituric acid), and weight lost, and water activity were observed. The results showed that the number of Williopsis saturnus var. saturnus stayed stable for 84 days (8 logs CFU/mL) in the samples. Application of coating with and without yeast and only yeast solution reduced aflatoxin production in the peanuts samples approximately 1.7, 0.4 and 0.6 ppb comparing with control (water) during 84 days of storage, respectively. Oxidation of peanut samples were depressed about 40 and 60% by application of coating and coating containing yeast cells comparing with control at the end of storage, respectively. Both water activity and weight lost at all film groups slightly increased. Therefore, the results showed that application of edible coating with the yeast cells has potential to use in industrial base to prevent aflatoxin production in peanuts.
Parul Thapar
Indira Gandhi National Open University, India
Title: Application of biosensors for detection of contaminants in milk and milk products
Time : 15:05-15:20
Biography:
Parul Thapar is Research Scholar in Dairy Science and Technology from Indira Gandhi National Open University, New Delhi, India. She has contributed in Food and Health Sector in the form of published papers and articles. She has also achieved Young Scholar Award and Young Achievers Award from BRICPL Pvt. Ltd., India for her contributions in Science and Technology. She is currently working as Field Investigator at Maulana Azad Medical College, New Delhi, India in an ICMR Project on “Efficiency of child-to-child approach in preventing childhood injuries and their consequences”.
Abstract:
Statement of the problem: There are certain thermoduric organisms in milk and milk products which survive pasteurization temperature. Also, besides pasteurization, there are certain pathogenic organisms which gets activated during storage and packaging of milk products. Therefore, it is very necessary to keep the quality and nutritional aspects of food safe to consume it for the human being before it gets packaged. The advancement in technology is being carried out to in dairy industry, which has emerged out with the use of biosensors for determining the quality of milk and milk products. Biosensors are devices that can combine a biochemical molecule with a physical signal that can be translated into an indication of the safety or quality of food. These devices are evolving as substitutes for the existing conventional techniques. There are different types of biosensors depending upon the substrate to be used. Example- Electrical biosensors, Immobilized based biosensors, Optical biosensors, nano-material based biosensors and microbial biosensors.
Methodology & Theoretical orientation: Various techniques like enzyme substrate assay and spore based biosensors have been experimented at various research institutes in India for detection of thermoduric organisms like Micrococci spp, Listeria spp and broad-spectrum antibiotics in milk and milk products.
Findings: These technologies have come out to be cost-effective, highly-sensitive, wider applicability and a good replacement for the existing conventional techniques.
Conclusion and significance: India is the major producer and exporter of milk and milk products. The use of this technology can be beneficial for keeping the nutritional and quality aspects of milk and other food products. This will also reduce packaging costs and there is a potential in carrying out online monitoring of raw materials, trace compounds, vitamins, flavours, additives and contaminants in future.
Javeria Samad
Habib University, Pakistan
Title: Prevalence and sources of food borne infections attributed to drug resistant pathogens in street food of Karachi, Pakistan-A risk to public health
Time : 15:20-15:35
Biography:
Javeria Samad is a Biotechnologist by degree and has been working in Infectious Disease Research Laboratory (Molecular Biology Section) at The Aga Khan University, for the last seven years, mainly on Molecular Epidemiology of Enteric Diseases and Public Health aspects in pediatric population. She has a vast experience of Molecular Biology and Microbiology techniques and has been involved in multiple international research projects in collaboration with WHO, CDC, NIH, Bill & Melinda Gates foundation, University of Virginia (USA), University of Maryland (USA), University of Emory (USA). She was also part of biosafety activities and QC related activities in the research lab. Her work has been presented in some of the national & international conferences including IMED 2016, ASM Microbe 2016, 63rd ASTMH, Keynote symposia conference and IICE in April 2017. Currently she is enrolled in M.Phil./Ph.D. program as well as serving in an academic institute, Habib University (Karachi, Pakistan) as a Microbiology Researcher and Lab Instructor too.
Abstract:
Introduction: Foodborne illnesses are a major cause of public health concern in developing countries. In the metropolitan city of Karachi, Pakistan, street vended food is very popular, acting as a major contributing factor of gastrointestinal diseases mainly due to contaminated food and water. This qualitative study was carried out to assess the microbial quality of street food and to determine the factors contributing to foodborne infections.
Methodology: A brief survey about hygiene and sanitation practices, demographics, food storage conditions and location of carts was conducted with twenty-six randomly selected street food vendors from average socioeconomic areas of Karachi, Pakistan. Hand washing water samples were collected in sterile bottles as a sample. Food items included lentil burger, chick peas, juices, fried items, meat curry items, ice-cream and fruit cream. Vendors were educated about basic hand and food hygiene practices and provided with soap bars. Samples (n=52) were enriched and inoculated on differential and selective media, results were recorded based on growth, morphology and biochemical tests. Antibiotic susceptibility was measured using Kirby Bauer test.
Results: Survey revealed that most of the vendors were shabbily dressed and were illiterate without any food hygiene training, showing poor practices. All the hand washed water samples and food samples were contaminated with known human pathogens. All samples exhibited high resistance to antibiotics, Amoxicillin (80%) being highest, Augmentin (45%), Gentamicin (39%), Ceftazidime (34%), Ceftriaxone (11%) and Ciprofloxacin (35%). MRSA exhibited additional resistance to Vancomycin, Oxacillin and Linezolid. Organisms isolated from food showed more resistance than organisms isolated from hands.
Conclusion: This study provides evidence about deteriorating public health conditions due to unrestricted dissemination of food borne pathogens. Presence of drug resistant organisms should be a great concern for the government authorities, suggesting the need for proper implementation of food safety measures.
Grettel Aviles Sayas
The Universidad Veracruzana, Mexico
Title: Nickel and vanadium toxicity in plant growth promoting microorganisms
Time : 15:35-15:50
Biography:
Grettel Aviles Sayas is currently a graduate student in Agricultural Science at the Universidad Veracruzana, Mexico. He has experience in the fields of biotechnology and microbiology, having worked in the Microbiology laboratorie of the Biological Research Institute of the Universidad Veracruzana, Xalapa, Mexico, in the department of in vitro culture of INBIOTECA, Xalapa, Mexico, and in the laboratory of Microbiology of the Colegio de Postgraduados, Texcoco, México. He worked in projects of genetic improvement in plants, and of degradation of hydrocarbons by bacteria. He did my degree in Biology (2015) and He is currently doing his Master's degree in Agricultural Sciences, both at the Universidad Veracruzana. At the moment he is working on the realization of two scientific articles.
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 NiSO4 (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.
Nadeem Hafeez
Center for Excellence in Molecular Biology (CEMB), Pakistan
Title: Hertrologous expression of uncharacterized wild type and mutated Universal Stress Protein-2 (USP-2) gene from Gossypium arboreum-FDH-171 confers osmotic and salt resistance to Pichia pastoris and Escherichia coli
Time : 15:50-16:05
Biography:
Nadeem Hafeez is currently working as a Researcher at Center for Excellence in Molecular Biology (CEMB), Lahore, Pakistan. He has expertise in gene editing, cloning and plant transformation; have developed a first triple gene genetically engineered cotton variety of Pakistan, which confers significant resistance against biotic and abiotic stresses. He has utilized E. coli and yeast expression models to screen mutated genes before transformation in plants. He is 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 37-45°C, Cold 4°C). The expression of 1st mutant (M1usp-2) was noted as 8.3 fold under NaCl stress and 9.7 fold 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 2nd 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 3rd 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 1st and 3rd mutant forms of USP-2 may directly involve in stress adaptive mechanism or it might be functional as a signalling molecule to initiate stress mechanism.
- Poster Session
Location: Goya
Session Introduction
Ahmed Marroki
University Djillali Liabes, Algeria
Title: Heterogeneity in technological traits of lactobacilli isolated from Algerian goat’s milk
Biography:
Ahmed Marroki is a Research Professor full time in Department of Biology-University Djillali Liabès of Sidi Bel Abbès-Algeria. He obtained PhD in Microbiology at the Faculty of Sciences-University of Oran1-Algeria and is a Member in the Laboratory of Microbial genetic Group-" Bacterial consotium, zootechnics performance and sanitary prevention in poultry production" University of Oran1-Algeria. His academic and research background was focused on teaching courses and conferences for students and contributing in several scientific papers including, scientific articles in international journals and book chapter, participation in national and international conferences and congress. His current research focuses on characterization of lactic acid bacteria including, phenotypic and molecular identification, probiotic strain selection, production of bacteriocin. He has also a particular interest in microbiology diversity of human and animals ecosystems. Currently, he is reviewing manuscripts and papers in different high-quality journals, specialized in Biology, Sciences technology, Biotechnology, Agriculture and Microbiology; and member of the editorial board of international journal.
Abstract:
Lactobacilli are rods belonging to the Lactic Acid Bacteria (LAB), and includes: Gram-positive, catalase negative, non-motile, non-sporulating, facultative anaerobes, growing under microaerophilic to strictly anaerobic conditions. Recently, some research has been focused for selected strains from goat’s milk with a technological potential and safety, susceptible to be used as adjunct cultures in dairy products strains intended for the use in food systems as starters or probiotics adjunct culture should therefore be carefully examined following important criteria. This study is a widespread of our research that has of objective to identify and study in vitro the technological traits heterogeneity and their safety aspect of lactobacilli strains isolated from goat's milk samples collected in different regions of Algeria. Using API50 CHL system and 16S rDNA sequencing, 51 % of strains were assigned as L. plantarum, 34 % as L. pentosus, 7 % as L. rhamnosus and 8 % as L. fermentum. A large variability was noted for the acidifying capacity in skimmed milk after 6, 12 and 24 h of incubation. All strains expressed aminopeptidase activity against alanine-ρ-NA and leucine-ρ-NA at different levels. All strains were resistant to vancomycin and most of strains showed more susceptibility to β-lactam antibiotic. High susceptibility toward the inhibitors of protein synthesis was also observed. Minimum inhibitory concentrations data obtained revealed that isolates were susceptible to penicillin and chloramphenicol, and resistant to gentamicin and vancomycin. Minimum inhibitory concentration distribution of other antibiotics showed variability. The analysis of graphical representation of principal component analysis of technological properties of L. plantarum and L. pentosus strains showed diversity among the isolates. Finally, eight L. plantarum, four L. pentosus and two L. rhamnosus strains, could be good candidates as adjunct culture in dairy product in Algeria.
Alejandro Garrido-Maestu
International Iberian Nanotechnology Laboratory, Portugal
Title: Isothermal DNA amplification for Salmonella spp. detection and characterization
Biography:
Alejandro Garrido Maestu is a Research Fellow working at International Iberian Nanotechnology Laboratory, with the expertise in Microbiology and Molecular Biology. At present he is working in a project based on a novel DNA amplification technique (loop-mediated isothermal amplification) for foodborne pathogens detection, combining with microfluidics and gold nanoparticles. This project is expected to have a great impact in the food industry as it will allow fast and accurate detection of the main foodborne pathogens. He has participated in several research projects involving bacterial agents detection, back in Spain and in the University of Florida where he started his Postdoctoral training. He has authored 10 peer-review papers with an h-index of 8, along with 1 book, all focused on food microbiology and bacterial pathogens.
Abstract:
Statement of the Problem: The genus Salmonella continues to be a major health issue. In Europe, more than 90000 of salmonellosis cases were confirmed in 2015 (1.9% increase respect to 2014), being serovar Enteritidis and Typhimurium the most prevalent ones, representing 45.7% and 15.8%. Conventional culture methods are lengthy and time-consuming, for this reason, the development of new methods that allow early detection of these pathogens is of high interest. Molecular methods have the ability of overcoming the previous limitations, being DNA amplification techniques the most attractive approach. In this sense, in recent years isothermal DNA amplification techniques have gained a lot of interest over other more stablished techniques, such as PCR/ qPCR, as providing several added-on advantages (no need of expensive equipment, many alternatives for product detection, etc.) One of these techniques with increase interest is loop-mediated isothermal amplification (LAMP).
Methodology: A selection of the most appropriate genetic targets was done, based on previously published studies, being selected invA for Salmonella spp. detection, STM4497 and safA for Typhimurium and Enteritidis serotyping respectively. Specificity and sensitivity assays were accomplished to ensure correct performance, and finally were implemented as the detection strategy after a simple pre-enrichment and DNA extraction. These new methods were compared against qPCR.
Findings: The newly designed assays performed excellent in terms of specificity, but presented lower sensitivity than qPCR. This problem was overcome with a pre-enrichment step. The evaluation of the performance of the assays in spiked food samples (Turkey, chicken and egg) indicated that they may be implemented in routine food-testing.
Conclusion & Significance: The developed methodology is suitable for routine food analysis, and may be used, either in a sequential mode or for directly assessing the presence of a specific serovar. LAMP represents a valid alternative to conventional DNA amplification techniques, and will allow costs reduction.
Carmen Candel-Perez
Universidad de Murcia, Spain
Title: Occurrence of Clostridium difficile in edible bivalve molluscs in Spain
Biography:
Carmen Candel Perez is a candidate for a PhD in Food Science And Nutrition at Murcia University, Murcia, Spain. Her research interest involves the occurrence of C. difficile in food products.
Abstract:
Statement of the Problem: Bivalve molluscs are a frequent source of viral and bacterial pathogens due to, among other factors, their filtering nature. Toxigenic C. difficile has been isolated from seafood samples with potential implications of transfer to humans.
Purpose: The purpose of this study is to assess the occurrence of C. difficile as well as bacterial indicators (Salmonella spp, Escherichia coli and reducers) in edible bivalve molluscs in Spain.
Methodology & Theoretical Orientation: A total of 129 samples, consisting in 123 samples of mussels (Mytilus galloprovincialis) and 6 oysters (Crassostrea cornucopiae), were purchased from outlets in the city of Murcia (southeast Spain). The isolation of C. difficile strains was carried out using enrichment broth suplemented with sodium taurocholate followed by etanol shock, prior to the culture on a selective media. The identification was carried out detecting Tpi gene using molecular techniques. The isolation and enumeration of bacterial indicators were investigated according to the ISO norm.
Findings: C. difficile was isolated from 8.94% (11/123) of the mussels investigated, what supposes 20.8% (5/24) from Mediterranean Sea and 6.45% (6/93) from Northwest Atlantic. No C. difficile was detected in modified atmosphere packs or in cooked mussels. All oysters samples were negative for C. difficile. About the associated flora, Clostridium sulphite reducers appeared in 31% (40/129) of the analysed samples and E. coli appeared in 1.5% (2/129). Salmonella spp. was not present in any sample. These bacterial indicators were in accordance with the current legal requirements.
Conclusion & Significance: These findings indicate that edible bivalve molluscs could be a potential source of C. difficile, with a slightly higher isolation rates than other studies. The intake of raw or poorly cooked contaminated bivalve molluscs with its spores could represent a risk for human health.
Giovanna La Salandra
Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Italy
Title: Molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in southern Italy
Biography:
Giovanna La Salandra, Head of Scientific Research and Development of IZS PB, has her expertise in molecular microbiology applied in the field of food safety. She is Manager of many scientific projects in the field of food safety, funded by Italian Ministry of Health. She has published many papers in peer reviewed scientific journals as author and co-author. She has participated in national and international congress/conferences focused on food safety and hygiene food control.
Abstract:
Methicillin resistant Staphylococcus aureus (MRSA) is a significant and costly public health concern because it may enter the human food chain and contaminate milk and dairy products causing foodborne illness. The purpose of this study was to determine the occurrence and the characteristics (SCCmec-, spa-, multi locus sequence typing) of MRSA isolated from dairy food, with respect to the genes encoding Panton Valentine leukocidin (PVL) and ability to form biofilm. Nine (7.5%) MRSA strains were isolated from 120 samples of milk and dairy products produced in Apulia region (south Italy) from January to June 2017. Three different spa-types and STs were identified: the most frequently recovered spa-type and ST was t355-ST152 (55.6%), followed by t899-ST398 (22.2%) and t127-ST1 (22.2%). All isolates harbored the SCCmec type V (66.7%) or IVa (22.3%) and in one isolate (11.1%), ST398/t899, the SCCmec resulted was not detected. MRSA isolates with genotype t355/ ST152/SCCmec type V were PVL-positive, whereas t899/ST398/SCCmec type V and t127/ST1/SCCmec type IVa MRSA isolates were PVL-negative. All MRSA isolates were low grade biofilm positive using a semi-quantitative adherence assay by microtiter plate (MTP). The most common MRSA strain (t355-ST152) is a CA-MRSA clone associated to human infections, and reported in many countries, including Italy. MRSA ST 398 (t899) is considered an important livestock-associated lineage associated to several cases of infections of variable clinical relevance. MRSA t127-ST1 can cause bovine mastitis and is associated with pig carriage in Italy and other European countries. The detection of MRSA in food of animal origin is a potential health hazard related to some factors such as the number of colonizer/infected food handlers, the level of hygiene in food plants and transport systems, and the level of raw milk contamination; thus it is necessary monitoring food-producing animals and improving hygiene in food practices in order to reduce the microbiological risk to minimum.
Gizem Cufaoglu
Kirikkale University, Turkey
Title: Presence of Staphylococcus aureus and Shiga toxigenic Escherichia coli O157:H7 in raw meat in Agri, Turkey
Biography:
Gizem Cufaoglu is a Research Assistant in Kirikkale University Faculty of Veterinary Medicine, Department of Food Hygiene and Technology. She is interested in food microbiology, bacteriophages and biocontrol of foodborne pathogens. She is currently working on her PhD, titled ‘A study on the prevalence and characterization of Listeria monocytogenes in chicken neck skin, and biocontrol of the isolates by lytic bacteriophages on chicken leg food model’.
Abstract:
Staphylococcus aureus and Shiga toxigenic Escherichia coli (STEC) O157:H7 are both important foodborne pathogens around the globe. S. aureus can cause a wide range of infections from skin infection to life-threatening diseases including endocarditis, pneumonia, sepsis as well as toxin-mediated conditions such as toxic shock syndrome; on the other hand Escherichia coli O157:H7 can cause symptoms from mild diarrhea to severe hemorrhagic colitis (HC), hemolytic uremic syndrome (HUS), thrombotic thrombocytopenic purpura (TTP). In this study it was aimed to find out the prevalence and seasonal distribution of Staphylococcus aureus in 214 frozen raw meat (74 turkey, 70 chicken and 70 beef) and the prevalence of Escherichia coli O157:H7 in 70 raw beef with the characterization of the E. coli O157:H7 isolate by multiplex PCR. Out of 214 meat samples, 25.7% (18/70) of the beef, 11.4% (8/74) of the chicken meat, and 5.4% (4/70) of the turkey meat samples were contaminated with S. aureus. Out of 70 frozen raw beef samples, only 1 (1.4%) was identified as both shiga toxin 1 and 2 producing E. coli O157:H7 by the detection of stx1, stx2, eaeA, hly, and fliCh7 according to multiplex PCR analysis. Our findings showed that occurrence frequency of S. aureus was higher in frozen raw beef than in raw chicken and turkey meat samples and the highest prevalences were detected in autumn months. Although the prevalence of E. coli O157:H7 was low in beef, the presence of virulence genes, especially toxin genes remain a significant public health concern.
Biography:
Gulsah Karabulut graduated from Ankara University, Department of Food Engineering in 2011. She started her graduate studies in Sakarya University Department of Food Engineering. She is currently working as a research assistant at the Department of Food Engineering at Sakarya University. Work areas include food microbiology, edible films, biodegradable packaging, biocontrol applications.
Abstract:
The mold growth leading physical, chemical and sensory changing in food products and carrying health risks with toxin production is a main problem in the food industry. In our study, as an alternative solution, bioactive materials were developed by incorporating Williopsis saturnus var. saturnus at different concentrations (0; 3.2; 6.8 and 8.8 cfu/cm2) into whey protein concentrate (WPC) based edible films. Viability of W. saturnus var. saturnus in the films was investigated during the time of the storage. Moreover, the antifungal activity of the films incorporated with W. saturnus var. saturnus was also tested against Penicillium expansum and Aspergillus niger. The effect of the antagonist yeast occurrence in the film on physical properties and microstructural properties of the films were analyzed. The optical properties and the mechanical properties of the film samples were also observed by SEM and Instron, respectively. The results showed that bioactive WPC films containing 8.8 log cfu/cm2 W. saturnus var. saturnus were able to maintain more than 60% of the initial antagonist yeast population. In addition, the films incorporated with the antagonist yeasts reduced mycelium growth of P. expansum and A. niger on the medium by more than 29% and 19% at pH 4.5, respectively. Incorporation of more than 3.2 log cfu/cm2 antagonist yeasts significantly increased water vapor permeability of the films (P<0.05). Increased population of the yeasts also significantly increased percent water solubility of the films (P<0.05). Moreover, the distribution of the antagonistic yeasts in the films presented homogenous microstructure at SEM. Increasing concentration of the killer yeasts in the films presented a significant tendency to greenness and yellowness as well as lightness values (P<0.05). The concentrations more than 3.2 log cfu/cm2 of killer yeast population in the films significantly increased tensile strength but decreased percent elongation at break (P<0.05).
Horia Radid
National Institute of Hygiene-Rabat, Morocco
Title: Microbiological quality evaluation of raw cow’s milk taken from the farms in Sale, Morocco
Biography:
Horia Radid is currently working as a Faculty of Sciences, University Mohammed V Rabat, Morocco. She possess laboratory experiments in “microbiology and genomic biology, medical, bacteriological and microbiological of food, water and food hygiene analysis”. Her research work mainly focuses to evaluate the microbiological quality of foods and food hygiene in order to raise the consumer awareness and to establish in the whole country a successful information system for the investigation and monitoring of the diseases that have a feeding origin.
Abstract:
Statement of the problem: Among the most popular origins of diseases that have relation with feeding, we find, the perishable commodities and particularly the milk and its products especially during the very hot summer days. The matched methods for conservation of milk and the hygiene measures had never been respected.
Aim: The objective of this study allows estimating the microbiological quality of raw cow’s milk of 40 taken samples, at Sale, from four farms, during spring 2013.
Methodology: In all the samples that we analyzed, we looked for many micro-organisms, like the total aerobic mesophilic flora, the total coliforms and fecal coliforms, Escherichia coli, Staphylococcus aureus, Streptococcus β-hemolyticus, Listeria monocytogenes, Salmonella and Brucella abortus.
Findings: The synthesis of the obtained global results during the microbiological tests of the cow’s raw milk which is collected from farms, doesn’t show any specific fluctuations during all the way long of the trial period. The microbiological quality test is always the same whether it is qualitatively or quantitatively.
Conclusion: It is then necessary to create some effective control measures, in order to protect the health of the consumer. For the best milk quality, the dairy farmers must submit the most efficient hygienic methods.
Biography:
Ilknur Civelek graduated from Afyon Kocatepe University, Department of Food Engineering in 2013. She started her graduate studies in Sakarya University Department of Food Engineering. She is currently working as a food engineer at SAKVA. Work areas include food microbiology, edible films, biodegradable packaging, biocontrol applications.
Abstract:
One of the most important problems of Kashar cheese producers is the mold growth on the surface during storage. Williopsis saturnus var. saturnus killer yeast has been reported to have antagonistic effect on mold and yeast reproduction in many studies. In this project, to investigate antifungal effect of W. saturnus var. saturnus yeast, it was applied directly on the cheese or with cheese whey protein based edible coatings to prevent mold growth. The reason for the use of yeast cultivar with edible coating is to produce edible films that contain antifungal effects by encapsulating yeast. In our study, 50 g of cheese samples were coated with edible films based on whey protein with or without W. saturnus var. saturnus or yeast without coating at a concentration of 6.9 log cfu/g and dried for 1 hour. They were packed in vacuum and without vacuum packages, stored at 4°C for 56 days. The number of W. saturnus var. saturnus, total bacteria, lactic acid bacteria, molds and yeast counts were observed throughout the storage period. Water activity, color and pH of Kashar cheese samples were also observed. The results showed that in the 56 days of the storage period; total bacteria and lactic acid bacteria populations were not affected by coating application with yeast; however, the number W. saturnus var. saturnus yeast was increased by 1 log cfu/g. Application of W. saturnus var. saturnus was observed to slight decrease the growth of other yeasts and molds on the cheese samples. The color of all cheese samples did not change with coating and yeast application. The pH and water activity of the cheeses showed an increase. In a conclusion, application of whey protein coating containing W. saturnus var. saturnus on the cheese samples have a potential to inhibit mold growth during shelf life.
Biography:
Javeria Samad is a Biotechnologist by degree and has been working in Infectious Disease Research Laboratory (Molecular Biology Section) at The Aga Khan University, for the last seven years, mainly on Molecular Epidemiology of Enteric Diseases and Public Health aspects in pediatric population. She has a vast experience of Molecular Biology and Microbiology techniques and has been involved in multiple international research projects in collaboration with WHO, CDC, NIH, Bill & Melinda Gates foundation, University of Virginia (USA), University of Maryland (USA), University of Emory (USA). She was also part of biosafety activities and QC related activities in the research lab. Her work has been presented in some of the national & international conferences including IMED 2016, ASM Microbe 2016, 63rd ASTMH, Keynote symposia conference and IICE in April 2017. Currently she is enrolled in M.Phil./Ph.D. program as well as serving in an academic institute, Habib University (Karachi, Pakistan) as a Microbiology Researcher and Lab Instructor too.
Abstract:
Introduction: Foodborne illnesses are a major cause of public health concern in developing countries. In the metropolitan city of Karachi, Pakistan, street vended food is very popular, acting as a major contributing factor of gastrointestinal diseases mainly due to contaminated food and water. This qualitative study was carried out to assess the microbial quality of street food and to determine the factors contributing to foodborne infections.
Methodology: A brief survey about hygiene and sanitation practices, demographics, food storage conditions and location of carts was conducted with twenty-six randomly selected street food vendors from average socioeconomic areas of Karachi, Pakistan. Hand washing water samples were collected in sterile bottles as a sample. Food items included lentil burger, chick peas, juices, fried items, meat curry items, ice-cream and fruit cream. Vendors were educated about basic hand and food hygiene practices and provided with soap bars. Samples (n=52) were enriched and inoculated on differential and selective media, results were recorded based on growth, morphology and biochemical tests. Antibiotic susceptibility was measured using Kirby Bauer test.
Results: Survey revealed that most of the vendors were shabbily dressed and were illiterate without any food hygiene training, showing poor practices. All the hand washed water samples and food samples were contaminated with known human pathogens. All samples exhibited high resistance to antibiotics, Amoxicillin (80%) being highest, Augmentin (45%), Gentamicin (39%), Ceftazidime (34%), Ceftriaxone (11%) and Ciprofloxacin (35%). MRSA exhibited additional resistance to Vancomycin, Oxacillin and Linezolid. Organisms isolated from food showed more resistance than organisms isolated from hands.
Conclusion: This study provides evidence about deteriorating public health conditions due to unrestricted dissemination of food borne pathogens. Presence of drug resistant organisms should be a great concern for the government authorities, suggesting the need for proper implementation of food safety measures.
Jiseon Lee
Korea University, South Korea
Title: Anti-anxiety effects of lactic acid bacteria via regulation of gut microbiota and metabolic alteration in chronic stress-induced anxiety mice
Biography:
Jiseon Lee has a Bachelor's degree in Food Engineering from Dongguk University in Korea. During the Undergraduate course, she has prepared a thesis that measured the benzopyrine content in fried food. Currently, she is majoring in Food Microbiology in her Master's course at Korea University, and studying probiotics and diseases. Probiotics are being studied to prevent not only human diseases but also aging and prevention of aging. Research on probiotics is essential for the development of food engineering, food industry and food microbiology.
Abstract:
Background: The realization that stress alters brain-gut interaction which is known to play a critical role in health and disease, including neuropsychiatric disorders, is rapidly advancing. Here we tested whether fermented Maillard reaction product (f-MRP) treatment modifies behavior across domains relevant to anxiety, depression, cognition, stress response, and social behavior. Furthermore, the fermentation of MRP by Lactobacillus rhamnosus 4B15 synergistically enhanced the above activities compared to those of unfermented MRP.
Methods: C57BL/6J male mice were administered. In addition, casein, MRP, f-MRP or normal diet-treated mice were also exposed to chronic psychosocial stress, and behavior was assessed. Murine restraint stress (RST) is regularly employed in the study of behavioral and biological symptoms associated with depressive disorders. Compared to baseline, the mice exposed to grid floor housing changed the amount of time spent in the open field, rotarod, light & dark box, elevated plus maze, forced swimming, novel object recognition 24 hr and spatial recognition 24 hr behavior. The f-MRP treated mice had significantly longer immobility duration in the open field test and increased their time in the center zone episodes between Cas and MRP. Mice treated with f-MRP for more than 11 weeks exhibited significantly decreased anxiety in the 7 types of behavior test, but these mice exhibited similar level of anxiety in few behavioral tests.
Conclusions: Chronic stress promoted depressive-like behavior throughout the 8 weeks stress period and these depressive-like complications will bring about gut barrier dysfunction. These results suggest that glycation-induced modification of casein and fermentation correlated strongly with the enhanced functional properties. L. rhamnosus 4B15 strain reduces stress-related anxiety by normalizing behavior via a synergistic interplay with the MRP compound and/or metabolites contained in this product.
Judith Jimenez-Guzman
Metropolitan Autonomous University, Mexico
Title: Authentic cotija cheese as a functional food: Study of peptides released during ripening
Biography:
Judith Jimenez Guzman is a Food Engineer working at the Food Science Department of the Metropolitan Autonomous University in Lerma, Mexico. She has her expertise in dairy biotechnology, including aspects of enzymology, fermentations and dairy products as functional foods. Besides research work, she does collaborative research and consultant work for several dairy enterprises in Mexico.
Abstract:
Authentic Cotija cheese is an artisanal ripened cheese produced in the mountains of central Mexico, made with raw milk and ripened for at least 3 months; due to its quality, it was awarded best foreign cheese in the 2007 Cheese World Championship. Because of the wide variety of microorganisms fermenting Cotija cheese, several metabolites are produced during ripening that may affect consumers’ health, such as bioactive peptides. The aim of this work was to study bioactive peptide production during Cotija cheese ripening and the effect of weather conditions in peptide production. We studied cheeses made in 4 different regions with different microclimates, and followed ripening for 6 months to determine peptide production. Results showed production of peptides with molecular weights ranging from 1 to 6 kDa in all cheeses. Furthermore, when antihypertensive activity of cheese extracts was compared with captopril® a high inhibition of up to 97% was observed, demonstrating that bioactive peptides are produced during Cotija cheese ripening and that authentic Cotija cheese has functional properties that give it a greater added value.
Lila Boulekbache -Makhlouf
University of Bejaia, Algeria
Title: Essential oils composition, antibacterial and antioxidant activities of hydrodistillated extract of Eucalyptus globulus fruits
Biography:
Lila Boulekbache Makhlouf has expertise in the extraction, characterization and evaluation of the biological activities of natural substances for their use in the food and pharmaceutical industries. She is a Team Leader in the research laboratory: Laboratory of Biomathematics, Biophysics, Biochemistry, and Scientometry (L3BS) at the University of Bejaia in Algeria and responsible of the doctoral training: Bio-resources, Environment and Technology Agro-Food.
Abstract:
Statement of the Problem: Aromatic plants and their essential oils have been used since antiquity in flavor and fragrances, as condiments or spices, in medicines, as antimicrobial/insecticidal agents, and to protect stored products. Eucalyptus globulus plant is known for its richness on bioactive compounds such as essential oils, phenolic acids, flavonoids and hydrolysable tannins. Because of their multiple biological activities essential oils of E. globulus are used in medicine, perfume and food industry.
Methodology & Theoretical Orientation: The present study was undertaken: to determine the chemical composition of essential oils extract from Eucalyptus globulus (E. globulus) fruits, using gas-chromatography coupled with mass spectrometry (GC/MS) method and to examine their antibacterial effects against reference pathogenic strains: Staphylococcus aureus (S. aureus), Bacillus subtilis (B. subtilis), Listeria innocua (L. innocua), Escherichia coli (E. coli), Pseudomonas aerugenosa (P. aerugenosa), compared to that of two antibiotics tetracycline and gentamicin.
Findings: Twenty eight volatile compounds were identified, with the predominance of sesquiterpenes and oxygenated sesquiterpenes compounds (61.2%). The antibacterial activity shows an inhibition effect of essential oils extracts against all the tested bacteria with MIC of 3 and 4 mg mL-1. A bactericidal effect is observed, with MBC varying between 3.62 and 9.05 mg mL-1, which demonstrates the sensibility of all tested bacteria to the essential oils of E. globulus fruits.
Conclusion & Significance: The high yield and the results of antibacterial activity found for the studied essential oil extracts from E. globulus fruit made them good candidates for their use in pharmaceutical and food industries.
Maria J Cantalejo
Public University of Navarre, Spain
Title: Relationship between microbial counts and lipid oxidation during ageing process of foal meat
Biography:
Maria J Cantalejo is a tenured Professor in the Department of Food Technology at the Public University of Navarre in Spain. She has her expertise in developing new food products and optimizing non-thermal process conditions to improve preservation of foods. She believes that a change in people's standard of life could be considered, mainly in the case of underdeveloped countries, since they could obtain some life essential raw materials. Therefore, one of her main projects is to develop new high quality products, safe, with a high nutritional value, with no additives added and long lasting at room temperature. This approach is responsive to eradicate hunger from the world.
Abstract:
Statement of the Problem: Ageing is a very important process, the goal of which is for meat to become tenderer. Foal meat stands out especially because of its own tenderness just 24 hours post-mortem. The lack of reference values in aging process has led to different studies about the most appropriate management of foal meat during that period. However, the impact of ageing is inconclusive given the absence of a measurement methodology and reliable statistics. Therefore, the aim of this study was to determine the shelf-life of foal meat (Longissimus dorsi) aged at two different times (1 and 8 days outside of the carcass -T1 & T8-, at 4 ºC) and the effect of the slicing process on day 1, 3, 6 and 9.
Methodology: The microbiological analyses were determined according to their corresponding ISO norms. Findings: Based on the total mesophilic & psychrotrophic aerobic counts, Pseudomonas sp., moulds and yeasts, significant differences were found between samples T1 and T8, the shelf-life of foal meat T1 being 3 days. On the contrary, neither LAB nor enterobacteriaceae were affected by the aging period. There was a negative correlation between microbial counts and lipid oxidation depending on the aging period. In fact, lipid oxidation was significantly higher (p≤0.001) in samples T1 than those T8. Foal meat is rich in polyunsaturated fatty acids, which are highly prone to oxidation. Thus, ageing time is a direct cause of lipid oxidation and the no interaction between loin and steaks ageing showed that the trend in both kinds of meat was the same with or without a loin ageing.
Conclusion & Significance: Contrary to other species, a loin ageing process outside the carcass may not be necessary in foal meat. Consequently, a longer preservation time of foal steaks may be achieved, if the free oxygen is controlled.
Martina Bohacova
Gaziosmanpasa University, Czech Republic
Title: Characterisation of extracellular DNA in Listeria monocytogenes biofilm with the use of fluorescent techniques
Biography:
Martina Bohacova is a PhD student in Microbiology who is studying biofilm matrix dynamics in L. monocytogenes biofilms. Currently, she is gaining experience with fluorescence based techniques like confocal laser scanning microscopy, general statistic methods, image analysis and risk assessment in food microbiology.
Abstract:
Extracellular DNA (eDNA) is an abundant matrix component that protects biofilm from the environmental stress, facilitate the horizontal gene transfer and serve as source of nutrients. eDNA is also found in Listeria monocytogenes biofilm, but it is unknown to which extent its importance as a matrix component varies from strain to strain. This study aims to determine if strain-specific variations exist, and if the role of eDNA in L. monocytogenes biofilms is dependent on the conditions under which the biofilm was formed. Biofilm forming capacity of 10 selected Listeria monocytogenes strains originating from food of different phylogenetic lineages and serotypes was examined using crystal violet assay. The strains were cultured under static condition in nutrient rich environment at 37 °C, and in low nutrient environment at ambient temperature. Biofilm forming capacity was evaluated by crystal violet assay, and the 3D structure of biofilm was studied by fluorescence staining and confocal microscopy. Extracellular DNA was quantified by PicoGreen® staining and fluorometric detection. Significant differences in biofilm forming capacity were found between the culturing conditions. There was a positive correlation between extracellular DNA production and total biomass under both culturing conditions and the significant difference was shown in extracellular DNA release between the conditions. Although extracellular DNA concentration was affected by biomass, confocal laser scanning microscopy (CLSM) images revealed information about spatial distribution in biofilm. Extracellular DNA signal distribution was found to be influenced by strain and lineage.
Biography:
Radovan Cobanovic has his expertise in microbiological food safety analysis. During the ten years of work experience in the field of microbiological analysis of food he has participated in various activities related to the prevention of outbreaks by pathogenic microorganisms and improving the quality of production of various products. He went through all steps of microbiological laboratory from performing standard analysis to the development and validation of new lab developed methods. Now a days he is expanding his expertise in the field of virology, especially foodborne viruses Hepatitis A (HAV), Norovirus (NoV) genogroups I (GI) and II (GII) through participation in the opening of the department of virology analysis of food in SP Laboratory.
Abstract:
Tea drinks are popular so their consumption is growing all over the world and becoming part of many lifestyles. During harvesting, handling, storage and distribution, teas are subjected to contamination by various molds, which may be responsible for spoilage and production of mycotoxins. According to mentioned the goal of this study was to evaluate presence of toxigenic molds and their secondary metabolites − ochratoxin A (OTA) in black and herbal teas on the Serbian market. Analyzed samples were collected from various retail shops from January 2015 until December 2016. Total number of analyzed samples was 787 (336 samples of black teas and 451 samples of herbal teas) and they were tested on molds according to ISO methodology (ISO 21527-2) and identified by dichotomous keys. The most predominant molds detected were: Aspergillus, Mucor, Rhizopus and Cladosporium. Ochratoxin A (OTA) is produced by Aspergillus spp. (A. ochraceus, A. westerdijkiae, A. steynii, A. carbonarius, A. foetidus, A. lacticoffeatus, A. niger, A. sclerotioniger, A. tubingensis) so further analysis on OTA were conducted on samples in which were detected Aspergillus spp. by validated method (based on BS EN 14132). Aspergillus niger and Aspergillus flavus have been associated with black tea (56 samples). Aspergillus section Nigri, including Aspergillus niger, A. acidus, A. tubingensis and A. carbonarius were found in herbal teas (91 samples) available on the Serbian market. Production of OTA was quantified in mold isolate from black tea in 5% of samples (1, 2-4, 6 μg/kg). As far as isolates from herbal teas are concerned it was confirmed that 16% of A. niger isolates were able to produce OTA in quantity of 0.89-6.1 μg/kg. In order to minimize the health risks resulting from the consumption of teas contaminated by toxigenic mold and mycotoxins, it is imperative to monitor the presence of these contaminants prior their use.
Rui Feng Mao
Guangxi University, China
Title: Analysis of microbial harm in sugar cleaning area of sugar mill
Biography:
Rui-feng Mao, was born in 1963, Guilin. In 1983, he has graduated from the Department of biology, Sichuan University, and received a bachelor of science degree in microbiology. In 2011, he has completed his doctoral degree from the school of chemistry and chemical engineering of Guangxi University and received his Ph. D. in engineering. He is currently an associate professor of food science at the school of light industry and food engineering of Guangxi University. He has long been engaged in the teaching and research work of microbiology and biochemistry. The main research areas include microbial isolation, screening, utilization of renewable resources.
Abstract:
In Guangxi of China, GMP (Good Manufacturing Practice) have been introduced in sugar mills since 2005 to comply with the requirements of the government administration, the clean zone was defined, and physical isolation was enforced as a procedure in sugar production. However, the rate of microbial detection in sugars did not reduce as expected. In this paper, the microbiological investigation and microbiological harm analysis of sugar clean zone in three sugar mills in Guangxi were reported. The methodology of investigating microbial species isolated from sugar cane clean area and its surrounding environment in sugarcane mill, as well as the distribution pattern of environmental microorganisms combined with air flow analysis and microbiological analysis were introduced. The research results provide the basis for control theory and control technology in the analysis of microbial harm in sugarcane mill.
Sabrina Hellali
BIPEA, France
Title: Production of external reference materials in food microbiology
Biography:
Sabrina Hellali
Abstract:
BIPEA organizes regular proficiency testing schemes (PTS) in many analytical domains, including microbiology in food. In order to help laboratories with their regulatory requirements issues, BIPEA developed a production of external reference materials (ERM) for microbiology in food. These samples should allow laboratories to check the trueness of their results at any time, outside the regular P-tests. For that purpose, BIPEA produced stable and homogeneous samples of meat contaminated with Escherichia coli, Clostridium perfringens, Bacillus cereus, Staphylococcus aureus and Listeria monocytogenes, at levels between 102 to 104 colony forming unity per gram (CFU/g). To check the production, controls were performed by analysing some samples all along the production process at the beginning of the study and at regular intervals during a 6 month period. The homogeneity was checked by calculating coefficients of variation, which were inferior to 30% for all the analytical parameters. The stability was characterized by comparing means of three samples to the mean obtained at the beginning of the study. The samples produced were thus considered as being sufficiently homogeneous and stable to meet the requirements of ERM: the results of enumeration of the different micro-organisms present in the minced meat after 6 months of storage at (-24±6) °C showed a good stability, with a maximum deviation inferior to 0.5 log (CFU/g).
Stanley Kelechi Dike
Imo State University, Nigeria
Title: Antibiotic resistance and plasmid profile of bacterial pathogens isolated from street vended foods in Owerri, Nigeria
Biography:
Stanley Kelechi Dike completed his PhD from Nnamdi Azikiwe University Awka, Nigeria. He began his academic career with Madonna University Elele, Nigera. He is a peer reviewer of several reputable journals and has contributed more than 16 papers in reputed journals. He is currently an Academic Staff and a Post-graduate departmental coordinator at the Department of Microbiology, Imo State University Owerri. He is due for his Postdoctoral studies in May 2018.
Abstract:
Street foods are common in Nigeria serving the purposes of supplementing family income and meeting food demands of low income urban dwellers. However, these foods can become vehicles for transmitting food-borne pathogens due to poor processing and handling. This study is aimed at assessing the bacteriological safety of three foods [‘abacha’ (African salad), roasted pork and ‘ukwa’ (Treculia africana)] commonly vended in Owerri, Nigeria. Twelve samples [four each of ‘abacha’, roasted pork and ‘ukwa’] were purchased from different vendors in Owerri metropolis. Bacterial were isolated from the food samples by the pour plate method on nutrient, MacConkey, eosin methylene blue and Salmonella-Shigella agar plates and characterized using a combination of cultural/biochemical and 16S rRNA sequencing technologies. The antibiotic susceptibility test was performed for each isolate by the disk diffusion method. Plasmid profiles of isolates resistant to four or more antibiotics were determined followed by plasmid curing and a repeat of antibiotic susceptibility test to indicate whether resistance is chromosomal or carried on a mobile element. Thirty- one isolates comprising seven species [Acinetobacter baumannii, Bacillus sp., Enterobacter sp., Escherichia coli, Klebsiella sp., Kurthia gibsonii and Salmonella sp.] were obtained from the food samples. ‘Ukwa’ showed the highest microbial load and multi drug resistance was observed in 11 (35.5%) of the isolates, seven of which harboured plasmids ranging 0.5-48.5 kb. Plasmid curing improved sensitivity of isolates to all antibiotics except ampicillin and nalidixic acid. The presence of pathogens and multi drug resistant isolates in these street foods renders them unsafe for human consumption.
Tianxiang Yang
Korea University, South Korea
Title: Characterization of combi-CLEAs for biogenic amine detection
Biography:
Tianxiang Yang is currently working at Department of Food and Biotechnology, Korea University, Korea.
Abstract:
Biogenic amines (BAs) are a group of low-molecular mass organic bases produced through decarboxylation of free amino acids during microbial fermentation. Due to the toxicological effects of BAs on human health, it is necessary to monitor the levels of BAs in foods. In order to conduct the rapid and sensitive detection of BAs in foods, an enzymatic assay using copper containing monoamine oxidase (AMAO2) and flavin adenine dinucleotide containing putrescine oxidase (APUO) from Arthrobacter aurescens has been developed. To improve the stability and combine the activity of both enzyme, cross-linked enzyme aggregates (CLEAs) technique was applied, AMAO and APUO have been combined as cross linked enzyme aggregates (combi-CLEAs) using glutaraldehyde, as cross linker. Through the optimization of the formation condition, the combi-CLEAs were produced with a yield of 82% at a large scale preparation. Comparing to free enzyme, combi-CLEAs exhibited a broader pH range, pH 6-8 for tyramine, pH 7-10 for putrescine, with higher activity. The stability of combi-CLEAs was also improved by more than 2-fold compared to that of free enzymes. Kinetics analysis indicated that combi-CLEAs had the similar kinetic patterns with free enzyme. Although a reduced catalytic efficiency was detected in combi-CLEAs, the substrate inhibition was mitigated. The combi-CLEAs was further tested to determine the concentration of mixed BAs, the enzymatic assay result showed that the change of kinetics parameters has no effects on the measurement. Thus, enzymatic assay using combi-CLEAs could be utilized for the BA detection in foods.
Tinatin Elbakidze
Laboratory of the Ministry of Agriculture, Georgia
Title: A survey for grapevine leafroll-associated virus-1 and -3 in Georgian vineyards
Biography:
Tinatin Elbakidze has her expertise in Microbiology and Molecular Biology. She has PhD in Immunology and Applied Microbiology. She has been working in a field of bacteriophages studying application of bacterial viruses against the highly pathogenic bacteria. Nowadays as a member of Laboratory of the Ministry of Agriculture of Georgia where the wine industry is of high importance, she is focused on agriculture and evaluates best diagnostic approaches to apply for a better diagnostic tool for screening a large-scale number of samples to manage the strategy to eliminate the spread of GLD viruses in future perspectives. Her study is under development in the frames of her project: “Detection of Grapevine Leafroll-Associated Viruses: GLRaV-1 and - GLRaV-3 in Grapevine Cultivars Distributed in the East Part of Georgia”.
Abstract:
Grapevine leafroll associated virus 1 (GLRaV-1) and -3 (GLRaV-3) are the most widespread and dangerous causal viral agents of severe grapevine viral disease - Grapevine leafroll disease (GLD). Georgian vineyards according to its importance as a wine-producing region is focused on increasing the quality of wines produced from indigenous grape varieties (whiteberried cultivar-Rkatsiteli and red-berried cultivar-Saperavi), making them in a clean international style. Main objectives of this study were to define which member of GLD is presented predominantly in Georgian vineyards also Application of serological approaches for the surveillance of serological types of GLRaV-s for a better diagnostic tool for the analysis of large number of samples. Survey for the detection of GLRaV-1 and GLRaV-3 viruses was conducted from widely distributed grapevine cultivars (Rkatsiteli and Saperavi) growing in vineyards located in traditional wine-producing region Kakheti, the east part of Georgia. A total of 600 samples (300- Rkatsiteli and 300 Saperavi) were collected and analyzed (Das Elisa kits, Bioreba, Switzerland) from ten vineyards during 2016 and 2017 growing seasons. The study determined the incidence of both virus serotypes (34.2%) as either single or mixed infections. Virus profiles were determined for individual vines. Analysis showed that GLRaV-3 virus was predominantly distributed in Saperavi cultivar (20.7%) while in case of Rkatsiteli both viruses were presented (GLRaV-1-18%, GLRaV-3- 17.7%) equally. The overall analysis showed that GLRaV-3 virus was the prevalent and contributed 19.2% of total samples while GLRaV-1 virus was presented in 15% cases respectively. Mixed infections were found in 3% of samples. Annually testing of vines used for further propagation in nurseries might be recommended in order to be sure in quality of new plant material. Diagnostic procedures should be comprehensive to define whether infected vine is removed from the vineyard or virus free rootstocks are used for establishing new vineyards.
Yeonjeong Seo
Korea University, South Korea
Title: Effects of lactic acid bacteria on intestinal barrier and inflammation in chronic stressinduced mice
Biography:
Yeonjeong Seo has completed her Bachelor’s degree in Food Engineering from Dongguk University in Korea. Currently, she is pursuing her Master's degree in Food Microbiology at Korea University, and studying probiotics and gut microbiota. Probiotics are live bacteria and yeasts that are good for human’s health, especially digestive system. When people lose good bacteria in the body like after taking antibiotics, for example, probiotics can help replace them. They can help balance good and bad bacteria to keep the body working like it should. She is trying to figure out exactly how probiotics work for human body and diseases.
Abstract:
Background: The realization that stress alters brain–gut interaction which is known to play a critical role in health and disease is rapidly advancing. In a stress model, we aimed at evaluating the effect of fermented maillard reaction product treatment on intestinal barrier and inflammation. Moreover, the fermentation of MRP by Lactobacillus rhamnosus 4B15 synergistically enhanced the above effect compared to those of unfermented MRP.
Methods: C57BL/6J male mice were administered. In addition, casein, MRP, f-MRP or normal diet-treated mice were also exposed to chronic psychosocial stress, then tight junction protein and inflammatory cytokines are evaluated by H&E staining and q-PCR. Compared to baseline, the mice exposed to chronic mild stress changed intestinal barrier and inflammation response. The f-MRP prevented the increase in intestinal permeability induced by chronic stress and restored tight junction protein such as occludin and ZO-1 expressions to control levels. In addition, expression of inflammatory cytokines was changed compared to baseline.
Conclusions: Chronic psychological stress contributes significantly to symptoms of functional bowel disorders and can predispose patients to gastrointestinal infection. Chronic psychological stress can also induce visceral hyperalgesia. In animal models, psychological stress appears to alter gut barrier function. It is conceivable that f-MRP alters gut bacterial communities, preventing intestinal inflammation and impaired permeability induced by chronic stress. These results suggest that glycationinduced modification of casein and fermentation correlated strongly with the enhanced functional properties. And L. rhamnosus 4B15 strain reduces gut barrier disruption and inflammation via a synergistic interplay with the MRP compound and/or metabolites contained in this product.
Yong Zhao
Guangxi University, China
Title: Differentiation of Listeria monocytogenes strains base on multi complete gene sequence
Biography:
Yong Zhao obtained his Doctorate of Science in Microbiology at the Nanjing Agricultural University in 2005. In 2007, he was selected for the young pioneer project of Shanghai College of Science and Technology, and he was selected for the “ShuGuang project" in Shanghai in 2015. His main research areas are Molecular Ecology of Food Microbiology, Food Quality and Safety Risk Assessment and Food Quality Safety and System Biology Research. He has hosted more than 20 scientific research projects, including the National Natural Science Foundation of China, and participated in more than 10 scientific research projects.
Abstract:
The Gram-positive bacterium Listeria monocytogenes is the causative agent of listeriosis, a severe foodborne infection. A multi complete gene sequencing scheme was developed for accurate, and sensitive subtyping L. monocytogenes. Complete DNA sequences for six virulence genes (prfA, plcA, hly, mpl, actA, and plcB) and two housekeeping genes (prs and ldh) in eight American Type Culture Collection (ATCC) L. monocytogenes strains with seven serotypes were sequenced and analyzed. Results showed multiple DNA sequence alignment identified 8 (prs), 5 (prfA), 8 (plcA), 8 (hly), 8 (mpl),8 (actA), 7 (plcB), and 8 (ldh) allelic types and a total of eight unique sequence types. mpl was the most polymorphic gene and the phylogenetic tree based on its sequence differentiated four groups with high bootstrap values. The mpl and ldh genes appear to be good molecular markers for further epidemiological investigation of L. monocytogenes. A combined virulence gene and the housekeeping genes multi complete DNA sequences scheme will be a promising method enhanced discriminatory power for L. monocytogenes.
Biography:
Youngbae Noh has majored in Dairy Science at Gyeongsang National University and obtained another major the Dairy Microbiology in Graduate School of Gyeongsang National University. He was joined in Dairy Team at Lotte R&D Center in Korea in 2016. He is a senior researcher of Dairy Team and in charge of research about probiotic strain and developing yogurt and fermented food. His major topic is fermentation of lactic acid bacteria and he has developed many probiotic foods such as LB-9 (Lotte’s probiotics brand) yogurt.
Abstract:
The plant-originated probiotics are known as beneficial bacteria that lived in fruit or vegetable fermented food. They were also known that exhibits various biological effects and has higher stability in stressed conditions than probiotics derived from dairy products. In this study, many lactic acid bacteria that isolated from Kimchi were tested for their characterization including tolerance to acidic and bile acidic conditions, adhesion to intestinal cells. Finally, LB-9, two novel lactic acid bacteria, L. Plantarum LRCC5193 and LRCC5273 were selected for application to many food products such as biscuit, chocolate, wafers, and milk. Also, we report that both the LB-9 probiotics ameliorate DSS-induced colitis using a mouse model. Under our experimental condition, oral administration of probiotic LB-9 and milk product of LB-9 was performed twice in a day for 7 days and the mice were followed by DSS treatment in order to induce pathological colitis. We found that DSS-induced intestinal apoptotic death was reduced in the mice treated with LB-9 compared to control. In addition, reduction of the apoptotic death was correlated with inhibition of proinflammatory cytokine-induced apoptotic signaling pathways by LB-9 treatment. Especially, NF-kB-mediated activation of the apoptotic index is significantly down-regulated in the DSS-induced colitis mice with LB-9. The present study supports the rationale for treatment of LB-9 as an important strategy to improve IBD, particularly colitis.
Zhang Ruili
NNSFC Foundation, China
Title: Inhibitory activity of the lactic acid bacteria from Xinjiang dairy products on Alternaria
Biography:
Zhang Ruili has been engaged in research work of food microbiology and inducing resistance in fruit for more than 10 years. He has his expertise in inhibiting postharvest diseases in Xinjiang fruit and the possible defense mechanisms involved. He has published more than 10 papers published in Chinese.
Abstract:
Statement of the Problem: Alternaria is one of the main pathogenic bacteria of fruit black spot in postharvest storage. The huge economic loss in Xinjiang province has been resulted by Alternaria. There were many fermented dairy stuffs in Xinjiang, China. We first tried to utilize the local lactic acid bacteria(LAB) strains from fermented dairy products to control the local pathogenic Alternaria.
Methodology & Theoretical Orientation: Two strains of A. tenuissima isolated from the pear and apple in south Xinjiang and A. tenuissima CGMCC3.3546 standard strain was used as pathogenic bacteria to detect the inhibitory activity of 110 local LAB strains’ cultures on the MRS agar, and in stationary and shaked MRS broth. The antifungal substances in the cultures of LAB strains with high inhibitory activity were detected by UPLC-Q-TOF.
Findings: Three Lactobacillus pentosus strains and one L. paracasei shown high inhibitory activity on A. tenuissima Ap1, A. tenuissima Am1 and A. tenuissima CGMCC3.3546. The shaked broth of three LAB strains had the higher inhibitory activity on the three Alternaria stains than cultures on MRS agar and in stationary broth. The main antifungal substances in shaked broth of the representative LAB 86 strain included organic acids, esters, N-heterocyclic compounds, alkaloids, and benzene derivatives, they had molecular less than 1000Da and synergistic effect of antifungal activities on Alternaria.
Conclusion & Significance: The shaked broth of three local LAB strains from Xinjiang dairy products could be used to control the black spot disease caused by Alternaria in the postharvest.