Madridge Journal of Food Technology

ISSN: 2577-4182

2nd International Probiotics, Nutrition & Microbiome Conference

October 10-12, 2018, Amsterdam, Netherlands
Scientific Session Abstracts
DOI: 10.18689/2577-4182.a2.010

Dysbiosis of Microbiome and Probiotic Treatment in a Genetic Model of Autism Spectrum Disorders

Tabouy L1*, Getselter D1, Ziv O2, Maayouf R1, Karpuj M3, Tabouy T4, Werbner N2, Mizrahi H2, Nuriel-Ohayon M2, Koren O2 and Elliott E1

1Bar-Ilan University, Israel
2Microbiome Research Lab, Israel
3Bar-Ilan University, Israel
4Paris-Saclay University, France

Background: The gut microbiome may influence brain development and behavior, mainly through the modulation of physiological metabolism and the immune system. Recent studies have determined that the microbiome has direct effects on behavior, and may be dysregulated in neurodevelopmental conditions. Considering that neurodevelopmental conditions, such as autism, have a strong genetic etiology, it is necessary to understand if genes associated with neurodevelopmental disorders, such as Shank3, can influence the gut microbiome, and if probiotics can be a therapeutic tool. Methods/Results: Using 16S high-throughput sequencing, we have determined the gut microbiome community of the Shank3 KO mouse model, and itsrelative controls. In this study, we have identified dysregulation of several genera and species of bacteria in both the gut and colon of Shank3 KO mice, in addition to a sex-dependent dysregulation of the immune system. L. reuteri, a species with decreased relative abundance in the Shank3 KO mice, positively correlated with the expression of GABA receptor subunits in the brain. Treatment of Shank3 KO mice with L. reuteri induced an attenuation of unsocial behavior and a decrease in repetitive behaviors, in males and just decrease in repetitive behaviors in females, without affecting anxiety. L. reuteri treatment also induced an increase in GABA receptor expression in multiple brain regions, and affected serum immune system markers.

Perspectives: This study has confirmed that genetic differences associated with autism can induce changes in the microbiota profile. In addition, this study identifies bacterial species that are sensitive to an autism-related mutation, and further suggests a therapeutic potential for probiotic treatment.

After sustained her PhD in 2012 in molecular and cellular neuroscience and structural biology within the functional and adaptive biology unit of Paris Diderot-Paris 7 University, Dr. Laure Tabouy taught at the Faculty of Sciences of the Franche-Comté University, then went for a post-doctorate in Israel, working on the role of the microbiome and the Gut-Brain axis in autism and the establishment of a probiotic treatment, in the Molecular and Behavioral Neuroscience laboratory of Dr. Evan Elliott, in the faculty of Medicine in the Galilee of the Bar-Ilan University. She continues to work on the microbiota in France at the Neuroscience Institute of Paris Saclay in France, (NeuroPSI)

Effect of Probiotics and Prebiotics on Gut Integrity in Critically Ill Patients

Daniyah Alkhawtani

Prince Sultan Military Medical City, Saudi Arabia

Gut failure is common in ICU patients which characterized by lack of bowel sounds, regurgitation, vomiting, high gastric residual volumes (>500 mL/day), diarrhea, constipation, abdominal distension or GI bleeding. During critical illness, several factors might affect gut microflora that involve changes in stress hormones, gut ischemia, use of antibiotics and immune suppression, gut micrbiota, lack of nutrients and enteral feeding failure. Unfortunately, clinical evaluation of the gut function is difficult therefore gut dysfunction usually goes unrecognized related to poor clinical outcomes and high morbidity and mortality rate. Diet has the strongest effects on gut microbial colonization that could modify the profile of dominant species in human gut and offer different consequences of health. Recent data suggested to preserve or reestablish a healthy gut microbiota during and after critical diseases through targeted interventions such as probiotics, prebiotics, fecal microbial transplants (FMT), and synthetic ‘stool pillsʼ. This review developed to evaluate the efficacy of probiotic/prebiotics in critical ill patients. Probiotics offers many benefits to the host including effective in the treatment or prevention of acute gastroenteritis, GI dysfunctions, antibiotic associated diarrhea (AAD), certain pediatric allergic disorders, necrotizing enterocolitis and inflammatory bowel disease (IBD). Briefly, probiotics use in the ICU remains widespread and no definitive recommendation for the routinely probiotics use in critical ill patients. Further studies are required to define the dose, types and safety of pro- and prebiotics in critically illness.

Keywords: Probiotics, prebiotics, critically ill patients, Antibiotic associated diarrhea, Clostridium difficile-associated diarrhea, Ventilator-associated pneumonia.

Daniyah Abdullah Alkhawtani is a clinical dietitian at Prince Sultan Military Medical City, Riyadh, Saudi Arabia. She obtained Bachelor of clinical nutrition from King Saud University, College of Applied Medical Sciences, Riyadh in 2011. She began her Master of clinical nutrition in 2013. Furthermore, she obtained Diploma in Sport and Exercise Nutrition. She Participated as a speaker in the advance course for residentʼs level 3(2015), 5th Clinical and Community Nutrition Update in Riyadh, Saudi Arabia (2016), 8th International Conference of Clinical Nutrition in Dubai (2016), 6th International Conference and Exhibition on Probiotics, Functional and Baby foods. London, UK, October 2017 and in 1st Annual Scientific Meeting of Saudi Arabia Parentral and Enteral Nutrition (SAPEN) Madareem Crown Hotel. Riyadh, Saudi Arabia April 2018.

Mechanisms Underlying Anti-Diarrheal Effects of Probiotics in Models of Infectious and Inflammatory Bowel Diseases (IBD) Associated Diarrhea

Pradeep K Dudeja

University of Illinois at Chicago, USA

Introduction: Many Lactobacillus species have been successfully used in clinical trials to treat diarrhea in children. However, mechanisms underlying anti-diarrheal effects of probiotics are not well understood. Recent studies from our group have shown that certain lactobacillus and bifidobacterial species upregulated electrolyteabsorption in the intestine. Since Intestinal epithelial apical membrane NHE3 (Na+/H+ exchanger 3) and DRA (Down Regulated in Adenoma, a key Cl-/HCO3- exchanger) play key roles in mediating intestinal electroneutral NaCl absorption, we sought to evaluate the efficacy of Lactobacillus acidophilus (LA) in counteracting NHE3 and DRA inhibition and ameliorating diarrhea in a model of C rodentium infection and DSS induced colitis.

Methods: FVBN mice challenged with C. rodentium (1x109 CFU) with or without administration of live LA (3x109 CFU) were assessed for NHE3 and DRA mRNA and protein expressio, mRNA levels of carbonic anhydrase, diarrheal phenotype (assessed by colonic weight/length ratio), myeloperoxidase (MPO) activity and proinflammatory cytokines. For colitis studies, dextran sulfate sodium (DSS in drinking water for 7 days) was used to induce colitis in C57BL/6J mice.

Results: LA counteracted C. rodentium-induced inhibition of colonic DRA, NHE3 and carbonic anhydrase I and IV expression, attenuated diarrheal phenotype and MPO activity. Further, LA completely blocked C. rodentium induction of IL-1β, IFN-γ and CXCL1 mRNA and C. rodentium-induced STAT3 phosphorylation. Oral gavage of live LA also showed evident alleviation of inflammatory state by reducing the weight loss, decreased colon length and colon weight in both the models. Also the inflammation as well as the decrease in DRA mRNA and protein levels caused by DSS colitis was blocked by LA.

Conclusions: Our data provide mechanistic insights into anti-diarrheal and anti-inflammatory effects of probiotics in models of infectious and chemical injury colitis.

Dr. Dudeja is a Professor of Physiology in the Department of Medicine at University of Illinois at Chicago and a Senior Research career Scientist at the Jesse Brown VA Medical Center. His group focuses on pathophysiology of diarrheal diseases as it pertains to infectious & IBD associated diarrhea and to develop better therapeutic interventions. His recent studies have focused on defining the mechanisms underlying potential antidiarrheal effects of probiotics. He has published about ~220 original articles and has been supported by multiple grants from NIH and the Department of Veterans affairs. He serves as an Editor for “Intestinal Absorption” for Comprehensive Physiology journal and on many editorial boards including: Gastroenterology, Amer. J. of Physiol., Physiological reports and Cell. & Mol. Gastroenterology & Hepatology. He has also served on many peer review committees including National Iinstitute for Health, and Department of Veterans Affairs, USA. He has been honored by Mario Toppo Distinguished Scientist Award from the ASIOA (Assoc. of Scientists of Indian Origin in America) and is a winner of Takeda Distinguished Scientist Award of American Physiological Society for 2018. These awards honor life-time achievements of Dr. Dudeja in the area of gut-microbe interactions in pathophysiology of diarrheal diseases.

Saccharum officinarium (Sugarcane) Molasses Enhances TGF-β Secretion and FoxP3 Expression by Probiotic Yogurt Culture Bifidobacterium animalis subsp. lactis BB-12 Stimulated PBMCs from Patients with Ulcerative Colitis

Sheikhi Abdolkarim1*, Mirzapour Majeed2, Shakournia Abdolhosain3, Baharifar Vahid3, Baharifar Narges1, Aqamohammadi Nima4, Sheikhi Mehdi5, Nowroozzadeh Enayatollah2, Matinrad Mehdi2, Mousavinasab S. Nouraddin6 and Sheikhi Razieh7

1Dezful University of Medical Sciences, Iran
2Caruncaneco, Iran
3Ahvaz University of Medical Sciences, Iran
5Dezful University of Medical Sciences, Iran & Kazeroon Azad University, Iran
6Sari University of Medical Sciences, Iran
7Tehran University of Medical Sciences, Iran

Running title: Sugarcane molasses enhances TGF-β secretion and FoxP3 expression by B. lactis stimulated PBMCs of UC Patients.

Background: Ulcerative colitis (UC) is one of the inflammatory diseases of the gut with frequent bloody diarrhea leads to increased rates of anemia. Evidences indicate the immunomodulation disorders in the response to intestinal microbiota in UC. Although sugarcane molasses, rich in necessary minerals and vitamins could be a good support nutrient but its effect on immune system of UC patients is unknown. To determine how the immune system of UC patients responds to molasses this study planned.

Methods: Bifidobacteriumlactis (B. lactis) were cultivated on MRS broth. Peripheral blood mononuclear cells (PBMCs) of 12 UC patients separated by Ficoll-Hypaque centrifugation and co-cultured with UV killed bacteria and/or molasses in RPMI-1640 plus 10% FCS. The FoxP3 gene expression measured by real-time PCR. TGF-β and TNF-α were measured in supernatant of PBMCs by ELISA.

Results: Sugarcane molasses and B. lactis significantly augmented TGF-β compared to control (p<0.01 and p<0.001 respectively). The secretion levels of TGF-β by B. lactis plus molasses compared to B. lactis stimulated PBMCs was significantly higher (p<0.05) but the level of TNF-α after 2/4/12 h incubation of PBMCs with B. lactis plus molasses compared to B. lactis alone wasnʼt changed (p>0.2). The level of FoxP3 expression after treatment with molasses was increased significantly (p<0.05). Although FoxP3 expression after treatment with the molasses plus B. lactis was increased but it wasnʼt significant compared to control.

Conclusion: These data show that if sugarcane molasses added to B. lactis, not only would not increase the pro-inflammatory cytokine, TNF-α, but also augments the anti-inflammatory cytokine, TGF-β by PBMCs. Increasing the expression of the FoxP3 could be due to stimulating the activity of the Treglymphocytes. Therefore, sugarcane molasses could be a safe support to compensate the lost nutrients in UC patients.

Keywords: Sugarcane molasses, Ulcerative colitis, B. lactis, Immunomodulation, TGF-β, FoxP3, TNF-α

Dr. Sheikhi started his B.Sc. in Shahid Chamran University in Biological Sciences and then his master in Medical Immunology in Tarbiat Modarres University. He did his Ph.D. at Immunology Institute, Kiel, Germany and Department of Immunology, Faculty of Medicine, Shiraz University of Medical Sciences (UMSc). Currently he is the professor of Immunology, head of department of Immunology and Microbiology, Dezful University of Medical Sciences. He currently studying the effect of yogurt derived probiotics and some support nutrients on the immune response of ulcerative colitis (UC) patients.

Lactobacillus acidophilus W37 Supports Intestinal Health and Immune Function Function

Nicole de Wit

Wageningen University and Research, The Netherlands

Lactobacillus strains are interesting probiotic strains which might have immune modulatory and other beneficial health effects, but each strain can be different in its activity. Here we tested three Lactobacillus strains, L. acidophilus W37, L. brevis W63 and L. casei W56, for their modulatory effects towards small intestinal epithelial cells in vitro and whether the most potent strain L. acidophilus W37 could have immune supportive effects in neonatal piglets. Caco-2 cells differentiated to small intestinal epithelial cells were exposed to L. acidophilus W37, L. brevis W63 and L. casei W56 (107 CFU/mL) for 6 hours. During exposure, trans-epithelial electrical resistance (TEER) was measured to determine the integrity of the confluent epithelialmonolayer. To determine the effect of the Lactobacillus strains on gene expression and related canonical pathways, RNA of Caco-2 cells was harvested after 6 hours incubation hybridized to Affymetrix Human Gene 1.1 ST arrays and Ingenuity Pathway Analysis (IPA) analysis was performed. Additionally, the potential beneficial effect of L. acidophilus W37 on intestinal integrity was studied using Caco-2 cells challenged with Salmonella Typhimurium (STM) DT12. In neonatal piglets, L. acidophilus W37 combined with long chain inulin type fructan Frutafit®TEX! was tested to enhance efficacy of vaccination against Salmonella Typhimurium. Treatments were given daily via oral drenches as of day 2 after birth until sacrifice. Piglets were weaned on day 24 and vaccinated with a single dose of Salmoporc STM®. To analyze the effect on protection against STM, animals were challenged with this pathogen before sacrifice. Blood was sampled prior and post vaccination to quantify antibody titers and characterisation of immune cells.

In the Caco-2 cell model, L. acidophilus W37 enhanced TEER by 15% (p<0.01) after 6 hours, while the strains L. brevis W63 and L. casei W56 had almost no effect on TEER. Transcriptomic data also showed that L. acidophilusW37 induced much more pronounced effects then the other 2 Lactobacillus strains. Most genes modulated by L. acidophilus W37 were related to intestinal barrier function, inflammation and bacterial stimulation of the epithelial layer. Many genes involved in the regulation of tight junctions were upregulated (e.g. CLDN 3, -4, Rab13)which fits the change in TEER. L. acidophilus W37 seems to impact epithelialimmune interactions via NFkB signaling and the upregulation of markers of follicle-associated epithelium (FAE) indicates that L. Acidophilus W37 might support epithelial cell differentiation towards tissue with enhanced intestinalimmune sampling capacity. Challenging Caco-2 cells with STM showed a drop in TEER of 24% (p<0.01) compared to medium control. This drop was only 3.9% (p<0.05) in cells pretreated for 17 hours with L. acidophilusW37 compared to medium control, illustrating the beneficial effect of this strain during STM infection. In piglets, the combination of L. acidophilus W37 and Frutafit® TEX! enhanced antibody titters against STM. This positive effect of L. acidophilus W37 on vaccination efficacy is indicative for its immunomodulatory potential. These results demonstrate that L. acidophilus W37can have beneficial effects on intestinal barrier function and aninteresting immune supportive activity. L. acidophilus W37 is therefore potentially useful to support intestinal health to e.g. STM burden in society and immune health for those groups that have a low immune fitness.

Dr. Nicole de Wit is a Senior Research Scientist at Wageningen Food & Biobased Research with major expertise in gut physiology. At the department of Human Nutrition of Wageningen University, she previously explored the role of nutrition, and especially high fat diets, on intestinal physiology at the molecular level in various mouse models. In her current job at Wageningen Food & Biobased Research, she is involved in multiple projects that study the role of fibres and pre- and probiotics in gastrointestinal health using in vitro models as well as animal models (piglets) and human intervention trials. For instance, she studied the immunomodulatory effect of probiotic strains by exploring their potential to support vaccination efficacy. She is also involved in public private partnership (PPP) projects in which the role of nutrition is studied to improve quality of life of irritable bowel syndrome (IBS) patients and how personalized dietary advice on fibre intake can empower consumers to make the right choices for improved health or reduced health risk.

Probiotics Dedicated to Honeybees

Aneta A. Ptaszyńska

Maria Curie-Skłodowska University, Poland

Probiotics – live microbial food supplements – are believed to be beneficial for their consumers by improving intestinal microbial balance, protecting against gastrointestinal disorders, and digesting some food components. Probiotic strains dedicated to some group of organisms should originate from healthy specimens belonging to this group (Sararela et al. 2000). Nowadays, probiotics are omnipresent as functional food and medicaments. Unfortunately, strains isolated from one group of organisms might not turn out to be beneficial for another, thus probiotics dedicated to vertebrates are harmful to or have no beneficial effects on insects (Whitten and Coates 2017, Ptaszyńska et al. 2016, Johanson et al. 2014, Mirjanic et al. 2013). Digestive system of honeybees is mainly inhabited by Lactobacillus kunkeei, L.apinorum, L. mellifer, L. apis, L. mellis and Fructobacillus fructosus (Hroncova et al. 2015).

Lactobacilluskunkeei and Fructobacillus fructosus bacteria make up the bulk of the natural, beneficial bacterial flora of honeybees in the summer. Owing to their properties, they occur in any environment with considerable amount of fructose, e.g. pollen, fruit, grape must, etc. Consequently, they also occur in the hive environment. Furthermore, these bacteria are also able to thrive in the honeybee gut, which produces numerous beneficial effects: they help digest and absorb necessary compounds and microelements, protect against mild poisonings with xenobiotics, acidify their environment, which protects the gut from any developments of microorganisms that can be pathogenic for bees, e.g. Paenibacillus larvae that cause foulbrood, or Nosemacerenae fungi that cause nosemosis. They also have immunomodulation effects improving beesʼ immunity, strengthening the condition of whole bee families and prolonging beesʼ lives.

In winter, bees are devoid of contacts with their outdoor environment, also with naturally occurring probiotic bacteria. Hence a whole bee family, especially young winter bees, ought to be in touch with a probiotic.

Dr hab. Aneta A. Ptaszyńska, a lecturer at the Institute of Biology and Biochemistry, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Poland has been researching honeybees and their diseases. She has been doing experimental studies into synthetic and natural substances that could make up the basis for remedies improving beesʼ condition, fighting bee diseases, mainly fungal diseases, such as nosemosis. Dr hab. Ptaszyńska has been cooperating with numerous scientific centres in Poland and abroad and her studies have been the basis for three patent applications, two industrial implementations, and numerous scientific publications.

Health-Platform End-Points: Application of New High Throughput Screening Technologies

Joe Hart

YBM International Ltd. Oxford, UK

Functional ingredients identified for health, nutrition and aging platforms are correlated with positive outcomes including cognitive health. Screening of botanical extracts has identified classes of components with positive outcomes including modifications of the microbiome. Traditional broad-spectrum functional extracts contain several active components. Individually these may also act on multiple targets such that overall positive outcomes integrate a variety of underlying mechanisms. EFSA health-related ingredient marketing claims, now require that defined endpoint efficacy be demonstrated and that a plausible mechanistic link be established.

Inherent variability in potency of biological material require measurements of biological efficacy with a strong predictive capacity. Different sage extracts, traditionally used for anti-inflammatory and estrogenic properties, were was tested witha proprietary, high throughput screening tool, Chronoscreen, which integrates outcomes at a whole organism level using a C.elegans nematode-based technology. It was configured with an array of endpoints indicative of cognitive health benefits. This enabled quantitative differences to be observed and to rank performance against established pharmaceutical actives.

In response to sage extract administration, gene expression studies allowed identification of up and down regulated genes and their associated pathways. Comparison with responses to Donepezil(a cognitive health therapeutic), allowed identification of differentially expressed genes and potential test dosages for human clinical trialconfirmation studies.

Further mapping to human health conditions suggested potential mechanisms of action consistent with neuro-transmitter modulation, longevity, autophagy and lipid homeostasis pathways and a new understanding of the biology of cognitive health.

A human clinical trial was carried out with 4 doses and a placebo given in a single daily dose. Cognitive health end point measurements over a 6-hour period following administration showed a time and dose-dependent significant positive differences with the placebo.

Conclusion: This quantitative screening approach showed predictive value for cognitive health performance and suggested potentially unanticipated modes of action.

Joe Hart completed his Undergraduate Biological Sciences (UEA); PhD program (Bacterial plant-pathogen virulence factors at UKC); Post-doctoral research (Secondary-metabolite molecular genetics in lactic-acid bacteria at NIRD, Univ. Reading): lead to microbiome-related role at Nestléʼs Swiss fundamental research centre. Across 17 yrs within Nestecʼs global R&D network, technical/marketing roles: infant nutrition; first validated pet food prebiotic claim; new functional ingredients. Identifying and consolidating ingredientʼs strategic benefits for Health and Nutrition platforms (human / animal / aquaculture) and production: Joe Hart has an experience in global product management (Daniscoʼs $35m natural preservatives / probiotics portfolio) and independent, ingredient-related strategic & business development consulting (together totaling 18yrs), and now leading YBM International Ltd. team.

Microbiota Composition of Dadih – A Traditional Fermented Buffalo-Milk of West Sumatra

Surono I S1* and Venema K2

1Bina Nusantara University, Indonesia
2Maastricht University – Campus Venlo, The Netherlands

Dadih is an Indonesian traditional spontaneous fermented buffalo-milk, produced in West-Sumatra, which is nutritious and has health-benefits. The mechanism of action behind the health-benefits is largely unknown, but several probiotic strains have been isolated from dadih, which may contribute to its health properties. To identify the composition of its microbiota, Two artisanal dadih samples (n=8) were collected from four producers. The raw buffalo-milk used for fermentation was either pasteurized (n=4) or not (n=4), and back-slopping was used as a starter-culture (n=5) or not (n=3). DNA was extracted from each sample in duplicate and the microbiota composition was determined by 16S-rRNA-gene amplicon-sequencing of the V3-V4 region. PCoA analysis showed clear separation of the samples by producer, but no separation due to pasteurization or use of back-slopping. Lactococcus (52%-83%) predominated in all samples, followed by Klebsiella (5%-26%), and Lactobacillaceae, Bifidobacterium (particularly high (~18%) in the non-pasteurized, back-slopped product from Palupuh), Streptococcus and Leuconostoc. Back-slopping practice correlated significantly with higher abundance of Lactobacillaceae, Pediococcus, species of the order Burkholderiales, and Serratia, but with lower abundance of several other Enterobacteriaceae (including Klebsiella), Streptococcaceae, Staphylococcus and Brachybacterium. Pasteurization was not significantly correlated with presence of certain members of the final microbiota. Taken together, fermentation results differ significantly from producer to producer and back-slopping practice would be advisable.

Keywords: Dadih, microbiota, fermentation, back-slopping, pasteurization, buffalo milk

Ingrid S Surono, is the Head of Food Technology Department, Faculty of Engineering at BINA NUSANTARA University, Jakarta, Indonesia. She earned MSc in Food Processing from Asian Institute of Technology (AIT), Bangkok-Thailand, in 1984. PhD was obtained from The United Graduate School, Gifu University-Japan, in 1996. She dedicated herself to probiotic and prebiotic research since 1992, and her research interests are in child nutrition, functional foods and food safety. She is co-promoter of 15 PhD students from Post Graduate Programs at Faculty of Medicine Airlangga University, Faculty of Medicine University of Indonesia, Faculty of Human Ecology Bogor Agricultural University, and Faculty of Medicine Airlangga University and of Andalas University. She has published 8 books, and wrote 2 chapters in text book published by CRC and Taylor Francis, and wrote 3 chapters in Encyclopedia of Dairy Sciences. She is an author of Probiotic, Microbiome and Functional Foods, published in 2016 in Indonesian. In 2011, Science and Technology Award have been granted to her from Toray Foundation. In 2012, she was nominated as ten best researcher of RISTEK KALBE SCIENCE AWARD 2012. She is President of Indonesian Scientific Society for Probiotics and Prebiotics (ISSPP), and one of Scientific Members at Danone Institute Indonesia since 2013-2017 and appointed as Head of Food Technology Department, Faculty of Engineering, BINA NUSANTARA University, Alam Sutera Campus, Tangerang, Indonesia.

Periodontal Muscle Training can Strength the Periodontal Support Feet your Teeth

Nima Sabzchamanara

National medical university, Ukraine

Previous research on periodontal structure and function has shown a significant relationship between periodontal tissue and teeth. This study assessed dentistʼs beliefs about the relative efficacy of the health of periodontal tissue. A total of 505 patients in general pracrice were asked to respond to a list of 25 obligatory nourishment for a child while going to have the first teeth, for its effectiveness in dealing with patientʼs periodontal health especially include chewing hard food. They were also asked to select the three most effective nutrition for periodontal tissue. The imdices of patient perceived importance of the periodontal health were derived and each compared with actual effectiveness as determined from a sample of 250 patients.

Although the majority of patientʼs rated 18 of 25 nutrition as being very effective, there was no significant association between patient perceived nourishment effectiveness and actual effectiveness. The implications for patient training are discussed.

Nima Sabzchamanara has completed his dental study from National Medical University Kiev Ukraine. He is the student of first year of Residency program, in the Department for Therapeutic dentistry. He has published one abstract in a dental journal.

Tackling Malnutrition, Obesity and Diabetes in Asia through Public Private Partnerships

B. Kistner

Defugo Bio, Germany

Introduction: It is estimated that the worldʼs population will cap in the year 2100 at 11 bio people of which 90 % will live in Asia and Africa. The pressure on air, space, water and food supply will be immense. For a company in the food and beverage industry it is evident that business growth will come from Asia and Africa. Today both regions face nutrition related health issues, both a problem for society but also an opportunity for the F&B industry to develop innovate and healthy food solutions and join the public sector in Public Private Partnerships to develop product innovation, new business models and form alliances.

Background: Aiming to contribute to Sustainable Development Goals SDG 2 & 3 – improving nutrition and SDG 17 partnerships. Today approximately 2/3 of humanity is affected by malnutrition – the fact that people have no access to food variety. Most devastating effects are during the first 1000 days of a humanʼs life – from conception to the 2nd birthday. If a human being has no access to a balanced diet during this time the person will either suffer from under-nutrition, stunting and wasting, have impaired brain capacity or will be overweight. In most Asian countries 30 % of children are stunted – too short for their age.

Obesity is lowest in Asia, between 10 and 15 % of the population; however, the increase in Asia is the highest with almost 30 % in the last 4 years as a by-product of economic development. Obesity often results in other health problems like diabetes II, colon cancer or heart disease. The cost of obesity to the healthcare systems in Asia are up to 20 % of the total healthcare expenditure.

Another health problem in Asia is osteoporosis – it is estimated that one in three women in Asia over 50 will suffer from it – this is mainly due to low milk consumption and no exposure to sunlight.

Mission: The public debate tends to be dominated by single solution discussions, which often polarize groups, fan outrage and make constructive collaborations difficult. What is needed is a system change to facilitate healthy food choices for a population in general. We must engage in multi-sectoral dialogues between industry, academia, government and non-governmental organizations. Further each involved party needs to be ready to change, this is essential for development.

Multiple, complementary interventions are necessary to show results. Studies are required on how to implement such interventions in different geo-political, social, physical, economic and cultural context. Interventions not as a linear process but rather holistic as a more circular process involving continuous learning, adaption and mainstream transparency.

Public Private Partnership Process

  • Start with data
  • Identify shared interest
  • Design for impact
  • Mainstream transparency
  • Measure outcomes

Used Sources:
Hans Rosling TED talk
Mc Kinsey Global Institute Report 2014
Economic Intelligence Unit Report 2017

Bruno Kistner is the Director Strategic Alliances at Defugo Bioceuticals, Singapore. He has worked in the UK as export coordinator with Roche as Head of Logistics, Marketing Manager in Middle East Region & Head of Emerging Markets Region HNH (India, Eastern Europe, Middle East and Africa). He is the Member of the Board Wuxi NutriRice Ltd. China.

Modulation of Gut Microbiome - From Phenomenology to Dietary Recommendations to Tackle Obesity

Stoffer Loman

NutriClaim BV, The Netherlands

The potential role of intestinal microbiota in the etiology of various human diseases has attracted massive attention in the last decade. Experiments in animal models have produced evidence for a causal role of intestinal microbiota in the etiology of obesity and insulin resistance. However, with a few exceptions, such causal relation is lacking for humans and most publications merely report associations between intestinal microbial composition and metabolic disorders such as obesity and type 2 diabetes.

Dietary recommendations are generally based on epidemiological evidence of an association between a food, food component or diet with the prevalence of a disease. However for microbiome-targeted dietary recommendations, little or no epidemiological evidence is available. In fact, world-wide no microbiome-related dietary recommendations have been proposed.

Nevertheless, diet-induced changes in the microbiota have been shown to be associated with insulin resistance and development of diabetes. However, it is as yet an enigma as to which extend the altered microbiota is causally related to insulin resistance and diabetes or whether these changes in the microbiota are secondary to the development of insulin resistance and diabetes, the latter two induced by non-microbiota dependent pathophysiological mechanisms. Thus, the reciprocal relationship between the gut bacteria and these metabolic disorders remains a matter of debate. In addition, and in particular with respect to fiber, the health benefits of its consumption may be conferred even without the apparent involvement of the microbiome.

In particular, e.g. studies into the role of fiber in its capacity to modify the microbiota to confer a health benefit have until now solely relied on measuring the changes in the microbiota at the start and completion of the study. Therefore, changes in the microbiota by itself are not sensitive enough to provide for a causal explanation of the observed metabolic effects. Unless multiple time points are included in the study design, kinetic of changes in the microbiota, combined with simultaneously revealing the kinetics of gene, protein and metabolome expression, applying multiple -omics techniques, microbiome-targeted dietary recommendations will remain elusive.

Stoffer has acquired a BSc-degree in Tropical Agriculture at the Royal Dutch College for Tropical Agriculture (Deventer, the Netherlands, was trained a Nutritionist (MSc) at Wageningen University (1992) and has obtained his PhD in Medical Sciences/Immunology at the Academic Medical Centre of the University of Amsterdam/ (1998). Following a career in the food supplement industry as science communicator and health educator he founded NutriClaim in 2007 (, providing specialist services pertaining to the scientific substantiation of health claims made on food, and marketing authorization of Novel Foods in the EU. Currently, Stoffer is also Work Package Leader in the EU FP7-funded project “MyNewGut”.

Healing Autoimmune Diseases with Plant—Based Skincare

Paul Joseph Griffith

Vegan Skin, USA

Vegan Skin by Paul Joseph is an all natural, plant-based, e-commerce, skincare, company that specializes in healing and hydrating the skin and scalp. Founder and C.E.O. Paul Joseph Griffith was diagnosed with a rare, autoimmune, skin, disease known as Hidradenitis Suppurativa when he was 23 years old. There is no known clinical cure or effective treatment for the disease. It is also degenerative and believed to affect approximately 1-2% of the American population and possibly globally. Throughout his formative years, the disease progressed and eventually spread across half of his body. He was covered in boils from the waist to his ankles and contemplated ending his own life. He decided in that moment to figure out a solution for himself driven by the research question, “How did our ancestors take care of themselves before modern medicine and technology?” Armed with this driving question, for four years he researched the healing properties of plants, focusing on their microbiology, and created three, signature, products for the skin and scalp. The products assisted in his healing and as a result became popular amongst friends and family members. Before long, the products became popular amongst customers across America with reports of assisted healing with autoimmune diseases such as eczema, psoriasis, dermatitis, folliculitis, cystic acne, chronically dry skin, rashes, and more. Thus, the focus of the oral presentation will be the intersection of the following:

1. The power of self—healing
2. The market for plant—based, natural, and wholistic solutions for skin and health
3. Plant microbiome and autoimmune disease treatment

Paul Joseph Griffith has completed his Masters at Tufts University, serves as the C.E.O. of Vegan Skin by Paul Joseph, is a part time professional dancer, a former public high school teacher and lives a busy vegan lifestyle in Washington D.C., U.S.A.

Fermented Plant and Seaweed Products – Prebiotics, Probiotics and Bioactives

Søren Kjærulff

FermBiotics ApS, Denmark

FermBiotics have developed several fermented plant and seaweed products for animal and human health. The products consist of prebiotics, probiotics and bioactives from the fermented plant and seaweed material. Metabolomic studies of the products have revealed several very health related metabolites and bioactives. We have demonstrated very good in vitro anti-microbial and anti-inflammatory activities of the fermented material. FermBiotics has performed several intervention studies in animal species and performed microbiome analysis of the gut bacteria and results have shown significant microbiome modulation. An additional pig trial showed a high (58%) reduction of LDL-cholesterol in pigs given 4 % fermented canola meal diet compared with normal diet. FermBiotics will in collaboration with Silkeborg Hospital, Denmark in the EU project Macro Cascade run a randomized double-blinded matched cross trial using a dose of 5 g fermented canola and seaweed material in 2h of 2018. The clinical trial will be performed in N=80 human patients with mild-moderate state of Inflammatory Bowel Disease (IBD) and/or Spondyloarthropathy (SPA) including both gender.

Søren Kj ærulff, CEO of Fermbiotics and CTO Fermentation experts. He is the responsible for development of fermented plant and seaweed food and pet food products with microbiome modulatory activity. He is responsible for introducing the product to the market, investigate and develop its functionality, identify the regulatory requirements, as well as go-to-market strategies. Former Novozymes senior director of biopharma R&D responsible of anti-microbial and anti-inflammatory peptides and albumin half-life extension of peptide drugs. He is the former vice president of R&D at Pantheco and Santaris Pharma.

Bacteriocinogenic Lactobacillus plantarum Inhibit Adhesion of Gastrointestinal Pathogens onto Extracellular Matrix and Intestinal Cells

Sandipan Mukherjee* and Aiyagari Ramesh

Indian Institute of Technology Guwahati, India

Probiotic lactic acid bacteria (LAB) can be explored to develop a safe and niche-specific therapeutic regimen against bacterial infections based on their ability to thwart adhesion of pathogens onto intestinal cells. The present investigation reports the potential of native Lactobacillus plantarum strains to inhibit adhesion of model gastrointestinal pathogens, Enterococcus faecalis and Staphylococcus aureus on extracellular matrix (ECM) and cultured intestinal cells (HT-29 cells). A native L. plantarum CRA21 strain rendered significant inhibition of S. aureus MTCC 96 and E. faecalis MTCC 439 adhesion onto ECM molecules, collagen and mucin, with the highest inhibition of S. aureus MTCC 96 adhered on collagen observed in the exclusion mode (83.61%) as compared to competition (62.4%) and displacement (38.58%) mode of adhesion assay. A dual-color flow cytometry (FCM) based adhesion assay indicated that in the exclusion mode, L. plantarum DF9 rendered notable inhibition of pathogen adhesion onto HT-29 cells, with the relative adhered population of E. faecalis MTCC 439 being 3.94% and that of L. plantarum DF9 being 77.56%. FCM along with principal component analysis (PCA) revealed that the native strains of L. plantarum DF9 and L. plantarum CRA38 could significantly affect both the adhesion process parameters kd and em of pathogen, similar to the standard probiotic L. rhamnosus GG. Interestingly, addition of the bacteriocin plantaricin A obtained from L. plantarum could reduce the viability of ECM-adhered pathogens. Host cell adhesion assays indicated that addition of plantaricin A on LAB and pathogen adhered onto HT-29 cells led to a prominent reduction in the adhered E. faecalis MTCC 439 cells (17.4%) as compared to L. plantarum DF9 (70.5%), which highlighted the benefit of using LAB bacteriocin for selective eradication of pathogen and minimal collateral damage. It is envisaged that the native probiotic L. plantarum strains can be used in conjunction with LAB bacteriocins as niche-specific anti-adhesion therapeutic agents against gastrointestinal pathogens.

Sandipan Mukherjee is a doctoral student in the Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, India. During his doctoral tenure, he has essentially ascertained the probiotic attributes of native lactic acid bacteria (LAB) and its secreted bacteriocin in inhibiting pathogen adhesion onto extracellular matrix and cultured human intestinal cells.

Proteomic Characterization of Bioactive Peptide from Endophytic Bacillus cereus

Rachna Pandey*, Laxmi Barapatre and Shivani Bhairamkar

Dr. D. Y. Patil Biotechnology and Bioinformatics Institute Tathawade, Dr. D. Y. Patil Vidyapeeth, India

Bioactive peptides or antimicrobial peptides (AMPs) characterization has received great attention in the recent past due to their application as therapeutic agents and also as food preservatives without any toxic effects on host. At present, food preservation is a serious concern for almost all countries across the world. A serious challenge to antimicrobial therapies is the rapid increase in antibiotic resistant infections. Therefore, an urgent need to develop other biocontrol agents is the need of the hour. Staphylococcus aureus is a major bacterial pathogen that causes clinical infection and foodborne illnesses. Food preservatives are widely used to reduce the risk of food poisoning. With growing consumer demand for natural preservatives to replace chemical compounds, new antimicrobial products of various origins are being developed. Endophytes from enormous number of diverse plants and environmental conditions are potential source for isolation of bioactive compounds. While a wide range of biologically active compounds have been isolated from endophytic organisms, they still remain a relatively untapped source of novel natural product. Endophytic microbes seem to fit perfectly into this natural warehouse, only a small part of which we have been able to tap in so far.

In this study, a Bacillus strain showed antimicrobial activity characterized by Kirby Bauer method of well/disc diffusion and soft agar overlay assay. The bacterial supernatant showed zone of inhibition after 16hr of incubation at 370C against Staphylococcus. The culture supernatant was precipitated with ammoniumsulphate (80% saturation) and column chromatography was performed with Sephadex G-50 followed by DEAE column purification. The purified fraction was used for characterization by Sodium Dodecyl–Polyacrylamide Gel electrophoresis (SDS –PAGE). The protein band thus resolved was taken for analysis by mass spectrometry (MALDI-TOF). Peptide mass fingerprint Analysis with MALDI showed bioactivepeptide of low molecular weight.

Keywords: Antimicrobia lpeptides (AMPs), Endophytes, antibiotic resistance, bioactive compounds, probiotics

Dr. Rachna Pandey is currently working as Assistant Professor at Dr. D. Y. Patil Biotechnology & Bioinformatics Institute, Pune, India. She earned her Bachelor of Science (B.Sc.) & Master of Science (M.Sc) at one of the premier institutes, Banaras Hindu University (BHU) India in 1993, and her Ph.D (Biotechnology) from the same University in 2001. In her thesis work she worked on Plant Growth Promoting Rhizobacteria Azospirillum sp. & itʼs Carbon Source Utilization. Her area of expertise has been Microbial Molecular Biology &Genomics. She worked as Research Associate (RA) in various Governments funded Projects in India.
After completing her PhD, in 2005 Dr. Rachna got Post Doctoral Fellowship from prestigious Department of Biotechnology (DBT) India and worked at National Chemical Laboratory (NCL) Pune. Where she worked on various aspects of carbon source utilization i.e catabolite repression (CR) & aspartic protease inhibitors (API).
In 2008, Dr. Rachna joined as an Associate Professor position at Assistant Professor at Dr. D. Y. Patil Biotechnology & Bioinformatics Institute, Pune, India, where she has been since that time. While at DYPBBI, she continued to work on Plant growth Promoting Rhizobacteria & also diversified towards the study of characterization & Bioprospecting of bacterial endophytes from medicinal & non medicinal and cereal plants. With constantly getting insight into bioprospecting of endophytes she characterized a bacterial endophyte with strong antimicrobial activity against different bacterial and fungal pathogens, addressing ever growing problem of antibiotic resistance among medically & agriculturally important bacterial & fungal pathogens. Recently, she is focusing her interest in characterization of unculturable bacterial diversity in some endemic plants. This work might help conserve plants which are on verge of extinction or are in rare or endangerd category. Dr. Pandey also work on Methylobacteriaum which play significant role mitigating methane levels emanating from deep water rice cultivation and coal bed mines. In her early years at DYPBBI she has characterized many PGPR strains including nonfluorescent Pseudomonas stutzeri an endophyte inhabiting Sunflower (Halianthusannus) roots.
Part of Dr. Pandeyʼs motivation for moving to characterization of Antimicrobial Peptide (AMPs) was because she got interested in addressing the issue of crop spoilage and heavy toll of crops due to pathogenic infestations. Also, already having realized the detrimental effects of use of chemical pesticides & chemical fertilizers in environment & aquatic habitat. Use of these deleterious chemicals is also carcinogenic and associated with other health hazards. Not only its use but also its production causes global warming and &unsustainable burden of the fiscal system. Biodiversity has always been her preferred areaof study. With 3,80,000 plants reported so far, they are promising candidates for exploring endophytic microbial diversity. As endophytes are poorly investigated group of microorganisms that represents an abundant and dependable source of bioactive and chemically novel compounds with potential for exploitation in a wide variety of medical, agricultural & industrial areas.
Dr. Rachna has over 30 refereed publications in International & National Journals.