International conference on Probiotics and Prebiotics October 30-31,2018
San Francisco, California, USA

Arthur C Ouwehand received his B.Sc. in biology and chemistry in Utrecht (the Netherlands) 1987, his M.Sc. degree (1992) in cell biology from Wageningen University (the Netherlands) and his Ph.D. degree (1996) in microbiology from Göteborg University (Sweden). Since 1999 he is Adjunct Professor in Applied Microbiology at the Turku University (Finland) and since 2004 he has been working for Danisco; now DuPont Nutrition and Health. He is the author of more than 250 journal articles and book chapters; he is the editor of four books on lactic acid bacteria and the intestinal microbiota and co-inventor on 15 approved patents.

Probiotics and to lesser extent prebiotics, have been widely used over the past 25 years. However, with the increased capabilities in microbiome research and associated fields such as proteomics and metabolomics, these product groups

have gained interest and new potential targets are being identified. There is also an increased understanding in the mechanism by which these ingredients work, which is expected to lead to the development of better targeted, more efficacious and more versatile probiotics and prebiotics. Microbiome techniques are also providing opportunities for identifying novel so-called next

generation probiotics, from new genera and species. Similarly, new prebiotic components will be identified; targeting different organisms then the usual lactobacilli and bifidobacteria. To understand where the pro- and prebiotic research fields are going, one can look data bases such as PubMed or in patent data bases; these, however, only allow us to look back at research that has been done. Doing a similar exercise with clinical trial registries may provide us a peek into the future; what intervention studies are being planned. We will see that some clinical end-points have reached maturity while other continue to attract attention and new end-points gain interest. In any case, the area of pro and prebiotics is very much alive and follows the technological developments. It is expected that this will translate itself into the products we will see on the market in the future.

Robert H Schiestl received his PhD from the University of Vienna, Austria at the age of 23. He did Postdoctoral work in Edmonton, Alberta, Rochester NY and Chapel Hill, NC. He was Professor at Harvard with the age of 31 where he stayed for 10 years. Since 18 years he is Professor at UCLA.

When our lab moved from Harvard to UCLA we found a huge difference in genetic instability and longevity in our Atm deficient mice after 5 years. When we changed the intestinal microbiota back to conventional microbiota we could reproduce the phenotype at Harvard. We tested Atm deficient mice for genotoxicity, genetic instability, DNA damage, inflammation markers, cancer latency and longevity and high throughput sequencing of the intestinal microbiota. Isogenic mice with different microbiota showed a four fold difference in life expectancy, a 4.5 fold difference in genetic instability and DNA damage. The onset of lymphomas was significantly 2.5 fold different. We sequenced the microbiota and found a Lactobacillus johnsonii 456 (LBJSupra) strain as dominant bacterial strain in the health beneficial microbiota. Just this bacterium by itself reduced genotoxicity, reduced inflammatory cytokines, induced anti-inflammatory cytokines and significantly reduced levels of cytotoxic T, CD3 and natural killer cells in the spleen, liver and blood. We also found similar differences in Trp53 deficient and even in wildtype mice. The underlying mechanism is due to inflammation promotion or suppression mediated by the intestinal microbiota. We did a clinical trial with this Lactobacillus strain that makes a great yogurt. 13 people took it for 7 days

and all of them had an increase of Lactobacilli in their feces, 7 of them had the same increase after 30 days and 4 of them 60 after days which is unique. This is because it expresses 16 proteins that bind to mucus which is also unique and it binds to human intestinal cells and thus inhibits binding of pathogens like Salmonella and pathogenic E. coli. It also expresses bacteriocins that kill pathogenic bacteria like Salmonella and pathogenic E. coli but not beneficial bacteria. LBJsupra is completely resistant up to PH 2 and pepsin, is at 100% viability in yogurt after 250 days in the fridge and can be scaled up for production. Inflammation is involved in most deadly diseases such as heart disease, cancer, neurodegenerative disease, inflammatory bowel disease,ulcerative colitis, Lupus, Crohn’s disease, Celiac disease, artheriosclerosis, arthritis, fibrosis, asthma and Diabetes and autism.

Amira A Ayad, Ph.D, is a Research Associate in the Food and Nutrition Sciences program at the North Carolina Agricultural and Technical State University-Center for Excellence in Post-Harvest Technologies. In this position, she taught several courses related to food analysis and food microbiology. Her research has focused on two main areas: Food Fermentation and Food Safety. Our research group focused on studying the fermentation processes of microorganisms like bacteria and how probiotic cultures as functional foods could influence the gut health and eventually human health. In addition to that, our research groups focus on studying natural antimicrobial properties against foodborne pathogens.

Bifidobacterium spp. and Lactobacillus spp. are probiotic bacteria have been incorporated in dairy-fermented products as live cultures to assist in maintaining the balance and health of human intestinal microflora. The aim of this study is to examine the ability of three Bifidobacterium (bifidum, breve and infantis) and Lactobacillus paracasei 441 strains to survive at

different (1.5, 2.0, 2.5 and 3.0) pH values and different (0.5 and 1.0%) bile salts concentrations for 3h, also to determine the stimulatory effect of L. paracasei 441 strain in combination with inulin on the viability of B. (bifidum, breve and infantis) strains in fermented milk during storage condition. Each strain of B. (bifidum, breve and infantis) were cultured with and without L. paracasei 441, then each treatment was split into three groups [0, 1.0 and 3.0 inulin (w/v %)] and incubated at 37°C for 3h. The viable bacterial counts of all strains were cultured in fermented milk during 15 days at 4°C was determined. The initial bacterial population reached 8.00 ±0.0 Log CFU/mL on an average. Our results showed that the bacterial populations of B. (bifidum, breve and infantis) strains in the control samples [without co-cultured with (L. paracasei 441 and inulin)] reached (7.45±0.0, 7.11±0.0 and 7.22±0.0) Log CFU/mL on an average, respectively. However, when the tested strains co-cultured with

L. paracasei strain 441 strain in combination with 1.0 % of inulin the bacterial population reached (8.67±0.0, 8.24±0.0 and 8.70±0.0). These results were indicated that there was a significant difference (P > 0.05) in the growth of B. (bifidum, breve and infantis) in fermented milk co-culturing with Lactobacillus paracasei 441 and Inulin. Therefore, our results indicated that the presents of L. paracasei 441 with 1% inulin was maintained and stimulated the viability of Bifidobacterium strains during 15 days at 4°C.

Cristina Bogsan is Pharmaceutical-biochemistry, formed by the Faculty of Pharmacy and Biochemistry of the Paulista University (1999). She holds a master's degree in Microbiology and Immunology from the Federal University of São Paulo (2002) and a Ph.D. in Sciences from the University of São Paulo (2012). Professor in the area of Food Technology in the Department of Biochemical Technology - Pharmaceutical, Faculty of Pharmaceutical Sciences, University of São Paulo since 2015, has experience in Microbiology and Immunology and Food Science and Technology, working mainly in the following subjects: B-1 cell, inflammation, fermented milk, matrix-mucosal-probiotic interaction, mucosal immunity, and probiotics. Development and functional characterization of probiotic products: in vitro and in vivo, development of novel functional foods.

With the increasing development of new functional dairy foods, there was a need to understand which mechanisms of action these foods present to confer health benefits. In addition to the nutritional effect of the food matrix and the

probiotic effect conferred by the fermented bacteria, the effect of diet-induced host epigenetic modulation could be evidenced. Within these epigenetic effects, interference by microRNA's inter-kingdom has particular attention. Epigenetics corresponds to a set of mechanisms that promote the regulation of gene expression by chemical modifications, such as methylation of DNA and chromatin, modification of histones and non-coding RNAs. These results in phenotypic change without alteration of the base sequence of the genomic DNA. Immune response regulators similar to human milk were identified in bovine and buffalo milk. However, food processing and probiotic interference in matrix composition may degrade and modify the presence and quantity of these miRNAs in the final product. Consequently, the modulation exerted by the consumption of the fermented milk. Among the biological processes modulated by miRNAs could be identified the differentiation of T cells, B cells, dendritic cells, and macrophages; Prevention of bone deterioration; Release of inflammatory mediators; Although this inter-kingdom regulation remains unclear. Even then, the benefits of probiotic fermented foods are greater than the sum of their individual microbial, nutritional or bioactive components.

Paulo Bastos has a degree in Biomedicine obtained at University of Aveiro (Portugal) and a degree in Biochemistry obtained at Biomedical Research Foundation, Academy of Athens (Greece), in partnership with University of Aveiro (Portugal). He has worked on a Neuroimmunology research group at Institute of Science and Technology (IST) in Vienna (Austria), a Microbiology research group at Institute Pasteur in Paris (France), a Proteomics Unit for the Academy of Athens (Greece), a Proteomics and Cell Signaling research group at the iBiMED, Institute for Biomedicine (Portugal) and a Clinical Pathology Laboratory at the Baixo Vouga Hospital Center (Portugal). He is the first author several peer reviewed publications. Currently, he is a PhD researcher at Institute Pasteur in Paris working on host microorganism interactions. The main project at the moment focuses on Mechanisms for the Dissemination of Bacteria and their Products from the Gut to the Brain and the Consequences Thereof.

Variations in gut microbiota composition and altered production of gut microbiota-derived products have been shown to influence a plethora of mammalian host processes, including gastrointestinal, immune and neuronal functioning and development. However, it is unknown whether and to what extent such perturbations can be mediated by peptidoglycan the major and fundamental constituent of the bacterial cell wall. We have thus set out to establish the in vivo uptake and distribution of gut bacteria peptidoglycan throughout the murine mammalian host. By employing fluorescently and radiolabeled bacteria peptidoglycan we have discovered a constitutive, physiological, uptake of gut bacteria peptidoglycan molecules from the intestinal lumen. We have subsequently established which organs are targeted the most and identified the brain as the major reservoir for gut bacteria peptidoglycan. The cortex was identified as the major reservoir of bacterial peptidoglycan, but the olfactory bulb is where peptidoglycan is concentrated the most. We are in the process of identifying which brain cell populations take up peptidoglycan the most. Most peptidoglycan taken up from the gut travels to distant organs via the bloodstream where we have established its proportional distribution (e.g. cells, exosomes, etc.). Other intriguing features were uncovered and are currently being studied, including the possibility for entero-hepatic recirculation and putative vehicles for peptidoglycan shuttling inside the mammalian host. We are in the process of demonstrating that gut bacteria derived peptidoglycan is functional in distant host target organs using an NFkB luciferase reporter mouse model. Alternative methods for in vivo tracking of gut bacteria peptidoglycan are in parallel being developed and herein presented.

Jong H Kim is a Research Molecular Biologist in the Agricultural Research Service (ARS), US Department of Agriculture, Albany, California. His research focuses on the development of intervention strategies for the control of mycotoxigenic and phytopathogenic fungi. He provides chemo-biological expertise, particularly in the identification of cellular targets, mechanisms of action and compound interaction and participates in resistance management in collaboration with producers, industry and academia.

Treatment of fungal pathogens such as Aspergillus fumigatus or producers of toxic secondary metabolites, viz., mycotoxins, is increasingly problematic due to the limited number of effective drugs or fungicides available for fungal control. Moreover, the expansion of fungal resistance to commercial drugs or fungicides is a global public health issue. For example, certain azole fungicides that are applied to agricultural fields have the same antifungal mechanism of action as clinical azole drugs. Such long-term application of azole fungicides to farms could provide selection pressure for the emergence of pan-azole-resistant fungal pathogens. Therefore, there is persistent need to enhance the effectiveness of conventional antifungal agents or discover/develop new intervention strategies. Current industry estimation indicates that development of new agrochemicals from discovery to the first sale requires 159,574 compounds to be screened on average, where the time and costs for this new development exceed 11 years and $286 million, respectively. We developed chemo-genetic approaches for compound screening to expedite the identification of new, safe antifungal agents. Screening United States Food and Drug Administration (FDA)-classified generally recognized as safe compounds led to the identification of chemicals targeting fungal antioxidant or cell wall integrity systems, which effectively inhibit the growth of pathogens. They possess antifungal, anti-mycotoxigenic or chemosensitizing capability to enhance the efficacy of conventional antifungal agents. Therefore, our methods can reduce costs, abate resistance and alleviate negative side effects associated with current antifungal treatments.

Shrilakshmi Desiraju received her Doctorate in Natural Product Chemistry from Vikram University, Ujjain M.P. She continued her Post-PhD research venture in peptide synthesis in NCL Pune & IISc Bangalore. She published 35 research articles and business articles till date. She worked on Pfizer projects in Cardio vascular division at NAEJA Pharmaceutical. Inc. Edmonton, Canada as Sr. Scientist and later she pursued Master of Business Administration (MBA) in Technology Commercialization at University of Alberta, Canada. After her MBA, she started R&D based nutraceuticals company Triphase Pharmaceuticals Pvt. Ltd. in Mysore, India in 2009. Under her supervision Triphase filed its 1st Indian and US patents in process development for thermostable probiotics in 2015.

Development of foods with adequate doses of probiotics at the time of consumption is a challenge, because several factors during processing and storage affect the viability of probiotic organisms. The food and beverage market is still beset by issues of strain stability, low product shelf life and challenges in the development of probiotic application technology. Several attempts have been made during the last few decades to improve the viability of probiotics in different food products during their production until the time of consumption. Major emphasis has been given to protect the microorganisms with the help of encapsulation technique, by addition of different protectants and by alteration of processing and storage conditions. Triphase’s Thermostable Probiotic strains (L. acidophilus & L. plantarum) that have the ability to withstand harsh manufacturing process, especially required during making Food, Beverage, Bakery and Pharma industries. These innovative strains especially have been developed using a proprietary process to ensure delivery of claimed dosage levels of probiotic. These strains don’t require any encapsulation. The present disclosure relates to thermo-stable strain(s) of micro-organisms, obtaining such thermo-stable strains and incorporating them into food products. These strains are able to survive high temperatures for longer periods of time and thus food products containing such strains are therapeutically effective and beneficial for general health.

Arnab Guha has done B.A.Sc. in Food Technology from Bhaskaracharya College of Applied Sciences, Delhi University & M.Sc. Food Science & Technology from CCS Haryana Agricultural University, Hisar. He is the Director of IIPL, a startup which provides techno-commercial life-sciences consulting services to food industry & agriculture. He is principal inventor in 5 applied patents. Before starting his entrepreneurial journey in Feb-2016, he had worked in Heinz, Britannia & Novozymes for ~10 years in domains like R&D, Technology transfer & Production. Earlier, he had undergone training for brief stints in Nestle, Coca Cola, Unilever (Kwality Walls) & Heinz.

Diverse culture of India brings out the best in the diverse ethnic cuisine of healthy fermented foods like Idly, Dosa, Dahi, Gundruk, Sinki, Nan, Kulcha, Bhatura, Dhokla, Lassi, Chenna, Handva etc. Idli dosa batter preparation process includes five major steps – Soaking (of ingredients like rice, black gram, millets etc.), Decanting (excess water), Grinding, Natural Fermentation, Packing and Storage (at 4-8°C). The science behind various physico-chemical property of idly dosa batter viz. bulk density, percent volume rise, pH, titrable acidity & fermentation time etc, is quite delicate. Processed packaged food even with natural ingredients is not considered as a fresh and healthy product. Fermentation is known to improve nutrition, palatability and digestibility. It suppresses food borne pathogens. Rishta Idli Dosa Batter from India is marching ahead on all these aspects. Our challenge becomes multifold when we work in super-tasty low-shelf-life fresh food product segment like Nutritious Packaged Idli Dosa Batter which requires cold chain in tropical climate. Being an affordable staple south Indian food, it’s an ideal vehicle to alleviate rural and urban malnutrition as well has improves digestion and mental well-being.