Day 2 :
- Track 1: Introduction to Probiotics
Track 2: Probiotics, Functional Foods in Economy and Business
Track 3: Current research and Future research perspectives on Probiotics
Location: SALA Auditorium
Cristina Stewart Bogsan
University of São Paulo, Brazil
Svetla T. Danova
The Stephan Angel of Institute of Microbiology associated with the Institute Pasteur International Network, Bulgaria
University of Sao Paulo, Brazil
Time : 12:15 - 12:35
Cristina Stewart Bittencourt Bogsan, Professor of Food Technology, Department of Biochemical Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, University of São Paulo and Researcher from TecLaFA, Technology of Dairy Functional Food and Analogues Lab, has graduated at Pharmacy and Biochemistry from Universidade Paulista in 1999, obtained her Master’s degree at Immunology and Microbiology from Universidade Federal de São Paulo in 2002 and has completed her PhD from Universidade de São Paulo in 2012. She has experience in Science and Food Technology, Microbiology and Immunology, focusing on Immunology and in Science and Technology of the Food, for the most part on Science and Technology of the Dairy Functional Food, acting on the following subjects: fermented milk, matrix-probiotic-mucosa interaction, Bifidobacterium animalis subsp. lactis, B-1 cells, immune stimulation and inflammation.
Diabetes mellitus has become a serious public health problem that affects millions of individuals worldwide. The World Health Organization predicts that 439 million people will have this disease in 2030, and Brazil was listed 5th of 10 countries estimated to have the highest number of people with diabetes, affecting approximately 12.7 million Brazilians in 2030. Kefir is a beverage made from milk that is fermented by a complex mixture of bacteria, including various species of lactobacilli and yeasts. It has been considered a probiotic due to its antioxidant and anti-inflammatory properties. The aim of this study was to investigate the effects of Kefir on the production of nitric oxide and oxidative stress and renal damage in STZ-induced diabetic rats. Diabetes was induced in adult male Wistar rats with streptozotocin (45 mg/Kg, iv). The animals received Kefir (K) or its vehicle 1.8 mL/day by gavage, starting from 5th day after induction of DM for 8 weeks. The animals were distributed into 4 groups (n=4 each): control (CTL); control Kefir (CTRK); diabetic (DM) and diabetic Kefir (DMK). Before and after treatment, blood and urine were collected for 24 hours to determine the thiobarbituric acid reactive substances (TBARS), NO, C-reactive protein (CRP), creatinine, urea and proteinuria. The data were processed statistically by one-way ANOVA with post-Newman-Keuls test (P<0.05). After sacrificing the animals, the renal cortex was removed for histology, oxidative stress and NOS evaluation. Comparing to CTL rats, DM rats had shown increased levels of glycemia, plasmatic urea, proteinuria, renal NO, superoxide anion, TBARS, and plasmatic CRP; also had been demonstrated a reduction in urinary urea, creatinine, and NO. However, DMK rats had shown a significant improvement in most of these parameters. Despite of lack of differences observed in the expression of endothelial NO synthase (eNOS), the expression of inducible NO synthase (iNOS) was significantly lower in DMK group when compared to DM rats, as assessed by Western blot analysis. Besides that the DMK group presented a significant reduction of glycogen accumulation within the renal tubules when compared to the DM group. These results indicate that Kefir treatment may contribute to better control of glycemia and oxidative stress, which is associated with the amelioration of renal function, suggesting its use as a non-pharmacological adjuvant to delay the progression of diabetic complications. Thus, Kefir may play a role in slowing the metabolic changes that contribute to DN.
Indian Institute of Technology, India
Title: Potential Clinical Applications of Probioticsration reduced progression of renal injury in STZdiabetic rats
Time : 12:35-12:55
Vikas Kumar completed his PhD from Banaras Hindu University, India, and Postdoctoral studies from School of Pharmacy, Texas Tech University, Amarillo, TX. He is now an Associate Professor of Pharmacology at IIT (BHU), Varanasi, India. He has published more than 100 papers in national and international journals and has been serving as an editorial board member of many scientific journals. He has mentored several PhD and MPharm students. He is recipient of many national and international awards and is a consultant to some pharmaceutical companies and research institutes. He has visited nine countries for his professional projects.
In the beginning of the twentieth century, the beneficial roles of nonpathogenic bacteria in the gastrointestinal tract were studied. A significant number of scientific studies have been apprised in support of the concept and evidences for the use of nonpathogenic organisms (probiotics) for clinical benefits. The cases of gastrointestinal diseases are growing around the world, and most of the gastrointestinal diseases occur because of alteration of intestinal microbiota ecosystem. Modification of the intestinal flora by increasing the predominance of specific probiotics would seem a reasonable approach of attaining a prophylactic or therapeutic effect against enteropathogens. The potential benefits of modifying the intestinal flora composition of certain high-risk groups, viz. premature infants, travelers, and children receiving antibiotics are emerging in modern days. An increasing number of healthcare professionals now routinely use condition-specific probiotic products against different diseases, either to treat them or for prophylactic purposes. Probiotics seem to play an important role in the gut that produces antibacterial peptides such as acidophilin bacteriocins to inhibit pathogenic microorganisms and improve the abnormalities of both the colonic flora and the intestinal microflora. Even though there was some evidence of the safety and efficacy of probiotics for treatment of diseases, recent clinical trials have proven its efficacies against many disease conditions.
University of Naples Federico II, Italy
Title: Lactobacillus paracasei CBA L74 prevents entrance of undigested gliadin peptides and rotavirus in Caco-2 cells
Time : 12:55 - 13:15
Objectives and study: Recent reports describe a role of probiotics as therapeutic approach for Celiac Disease (CD). Besides, rotavirus (RV) infections are described as a potential risk factor for CD development. Undigested A-gliadin peptides P31-43 and P57-68 are central to CD pathogenesis, entering in enterocytes in vesicular compartments by endocytosis and inducing an innate and an adaptive immune response respectively. Our aim is to test the effect of probiotic Heinz proprietary strain Lactobacillus paracasei CBA L74 (International Depository Accession Number LMG P-24778) and its supernatant on P31-43, P57-68 and RV entrance in Caco-2 cells to verify its protective effect. The effect of supernatant on ROS production in RV infected Caco-2 was also tested. Methods: We cultivated LP CBA L74, obtaining 108 CFU/ml, and its supernatant was obtained by centrifugation and filtration. Caco-2 cells were treated with LP CBA L74 or with its filtered supernatant at 37°C, CO2 5%, for 30 minutes, and then labeled P31-43/P57-68 or RV were added to cells cultures. We studied entrance of labeled peptides by fluorescence assay. RV entrance was assessed by direct immunofluorescence and the reactive oxygen species (ROS) production by dichlorofluorescein fluorimetric assay in Caco-2 cells infected with RV with or without bacterial supernatant. Results: LP CBA L74 inhibited both P31-43 (FI reduction: 67.28%, P<0.001) and P57-68 (FI reduction: 37,05%, P<0.001) entrance respect to control. LP CBA L74 supernatant was also able to induce decrease of both gliadin peptides entrance in Caco-2 cells (FI reduction: 49.38% and 29.67% respectively, P<0.001), indicating that this biological effect was due to some product included in LP CBA L74 supernatant. Supernatant significantly prevented RV entrance and ROS production (reduction of 56%, P<0.001) in RV infected Caco-2 cells, showing a potential protective effect in RV infections. Conclusion: LP CBA L74 and its supernatant reduce P31-43, P57-68 and RV entrance in Caco-2 cells probably acting on the endocytic trafficking. Moreover supernatant can protect Caco2 cells from RV mediated increase of ROS. This study is the first attempt to explain the molecular mechanisms of probiotic effects in the prevention of both RV infections and undigested gliadin peptides toxic effects
Shanghai Children’s Medical Center, China
Title: Characterization of the intestinal microbiome of Hirschsprung’s disease with and without enterocolitis
Time : 13:45 - 14:05
Li Hong is Associate Professor of Pediatrics, and the Director of the Department of Clinical Nutrition. She graduated from Fu-Dan University School of Medicine in 1994, following which she continued her pediatric surgical training until 1997. She has played a major role in the development of Nutrition support strategies for Children in China. She has consequently received many awards for her work, and published widely in peer reviewed journals. Her special interests include nutrition support in critical ill children, congenital GI malformations and intestinal rehabilitation, and microbiome research in children.
Hirschsprung’s disease (HD) is a congenital malformation of the gastrointestinal tract characterized by the absence of the distal enteric nervous system. Hirschsprung-associated enterocolitis (HAEC) is a severe life threatening complication of HD. The disease pathogenesis is still unclear, but evidences suggest that the intestinal microbiota may play important role in the development of HAEC. Because microbial abundance and diversity might differ in HD patients with enterocolitis, we sought to generate comparative metagenomic signatures to characterize the structure of the microbiome in HD patients with and without enterocolitis. Our experimental design is to enroll four HD patients (two with enterocolitis and two without enterocolitis). The microbiome was characterized by 16S rRNA gene, and the data obtained will be used to taxonomically classify and compare community structure among different samples. We found that the structure of the microbiome within HAEC patients differ from those without enterocolitis. Identifying the microbiome differences between HAEC and HD patients provides us a framework for future researches of determining the role of specific bacteria in inducing inflammation in HAEC. This study helps us to understand microbial contributions to the etiology of Hirschsprung associated enterocolitis, and provide a basis for early intervention of HAEC risk in Hirschsprung’s disease.
Selcuk University, Turkey
Time : 14:05 - 14:25
Nihat Akın has finished his PhD at Loughborough University Of Technology, Chemical Engineering Department-United Kingdom. He has been working as Professor for about fifteen years and been working as Head of the Department of Food Engineering in SelcukUniversity. He has published more than 100 scientific papers about milk products and microbiology.
Several fruit and vegetable juices have just been trying for probiotic vehicle. For this purpose pineapple juice, cabbage juice, cashew apple and carrot juices etc. were tried in the past. Study aimed to investigate the suitability of black carrot juice as Lb. Casei vehicle. Black carrots were provided by black carrot producers as a concentrate and it was diluted (1:10). Pasterization were applied to freshly prepared black carrot juices at 80°C for 20 min. For the purpose of decrease microbial population to be low the detection limit. 30°C and 48 hours were used for fermentation temperature and duration, respectively. İnoculum quantity were detected as 6,53 log CFU/ml. After two days of fermentation this number reached to 7,97 log CFU/ml. Optical density were determined before, after and at the end of the first day fermentation at 590 nm and by the way microbial growth was monitored during fermentation. After that storage was made at +4°C for 42 days. Analysis of viable cell count, color, Ph and titre table acidity were performed at each days.While pH was measured as 3,785 after inoculation, it was 3,695 at the end of the storage. Any loss was not observed for characteristic black carrot colour. Viable cell count of Lb. Casei were higher than 7.40 log CFU/ml during the storage period. This study shows that black carrot juice is convenient medium for L.casei and it could be used in functional food industry.
Selcuk University, Turkey
Title: Exopolysaccharide production of Lactic Acid Bacteria: Genetic background and future perspectives
Time : 14:25 - 14:45
Enes Dertli conducts research in the field of microbiology. His aim is to gain a better understanding of how probiotics show their beneficial effects and what are the molecular factors behind these effects. His research is focused on finding new strains for human consumption as probiotics and potential new starter cultures for food industry. After studying Food Engineering at Selçuk University, he finished his Master with the scholarship of Turkish National Science Council at same university (2008). Then he received a scholarship from Turkish National Education Ministry and moved to Institute of Food Research, UK where he finished his PhD in Molecular Biology and Microbiology (2014). At present he works as an Assistant Professor in Bayburt University, Turkey.
Several lactic acid bacteria (LAB) were shown to produce exopolysaccharides (EPS) which either forms a capsular layer around them or being directly secreted to the environment. These biopolymers were shown to be important for LAB for their putative roles in colonisation, adhesion, stress resistance, immunomodulation and host-bacteria interactions. They are also of considerable interest to food industry due to their essential roles in rheological properties of food products and food formulations. Bacterial EPS structure has a wide diversity among different LAB species due to the different sugar monomers in their repeating units and the glycosidic linkages present among these sugar monomers. Structurally, EPS are divided into two groups as homopolysaccharides which are formed by only one type of sugar molecule and heteropolysaccharides which consists of several sugar monomers such as glucose, galactose and rhamnose. Recent studies revealed that only single gene is responsible for the homopolymeric EPS production whereas eps gene clusters were shown to be responsible for the heteropolymeric EPS production. The structural properties of EPS are crucial for their role in colonisation and regulation of host responses as well as their role in food rheological properties. Several factors such as incubation temperature and time, sugar source and environmental conditions affect the levels of EPS production by LAB. This study will summarise recent findings in genetic and structural analysis of EPS production by LAB and their role in potential probiotic properties as well as recent applications in the food industry.
The Stephan Angeloff Institute of Microbiology, associated with the Institute Pasteur International Network,
Time : 14:45 - 15:05
Svetla T Danova is a molecular biologist with a strong background in microbiology and genetics of medically and industrially important groups of microorganisms (beneficial Lactic acid bacteria; antibiotic producing Streptomyces). She has graduated and completed her MSc and PhD thesis at the Biological Faculty, Sofia University, Sofia, Bulgaria. After the post-doc training in France, South Africa and Belgium she become a head of the laboratory “Genetics of Lactic Acid bacteria & Probiotics” at the Department of General Microbiology, The Stephan Angeloff Institute of Microbiology, a member of the International Network of the Institute Pasteur, France. She has published more than 50 articles on taxonomy, physiology and biological activity of lactic acid bacteria and probiotics and a student book “The Probiotics”.
Vaginal and urogenital flora plays a central role in maintaining both the wellbeing and illnesses of women. Recently, the lactobacilli that commonly are presented in the healthy vagina have attracted considerable attention. The understanding of protective role and probiotic properties of these beneficial bacteria is a promising way to advance the prevention, therapy and prophylaxis of urogenital infections (UTI) and related with risks. A complex study on Lactobacillus microbiota of healthy Bulgarian volunteers at childbearing age is presented. Using different culturable and non-culturable DNA methods, the Lactobacillus biodiversity were characterized. In addition, 35 newly isolated lactobacilli have been identified. Among the dominant species (Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus crispatus and Lactobacillus salivarius), several active strains, with a broad spectrum of antagonistic activity were pre-selected. In vitro tests in simulated vaginal fluid media showed the capacity of two vaginal strains to inhibit the growth and the biofilm-formation of uro-pathogenic Escherichia coli. The antagonistic and immunomodulation effects, established in vitro, were dose- and strain-dependent. The strain-specific antibiotic susceptibility, together with the adhesion ability to HeLa cell-line and the capacity for protective biofilm-formation, in conditions similar to those in vivo, were estimated as important part of mechanisms of stable colonization in the vagina. Moreover, some of vaginal lactobacilli combine biological activity with technological relevance. Obtained in vitro results implied that selected vaginal lactobacilli are appropriate as new multifunctional compounds for vaginal probiotic formula, with potential to maintain the urogenital health.
- Track 4: Probiotics Role in Control of Diseases
Track 5: Probiotics in Gastroenterology
Auburn University, USA
Auburn University, USA
Time : 15:05 - 15:25
Iryna Sorokulova, Professor of Microbiology received her MS degree in Microbiology from Taras Shevchenko Kiev State University (Ukraine), PhD and DSc degrees in Microbiology from the Institute of Microbiology and Virology, National Academy of Sciences of Ukraine. She has received a solid background in formulation and regulation of biological products during the training courses in Bureau of Biologics (Ottawa, Canada) and Massachusetts Biologic Laboratories (Boston, USA). She is an editorial board member of several reputed journals. She is the author of 82 refereed scientific publications, 4 books, and 23 patents.
Exposure to high environmental temperatures is considered to be one of the most deadly conditions, affecting human and animal health. Relatively small increase in internal temperature can lead to injury and even death. It was shown that heat stress results in a significant damage of the barrier function of the gut causing increased permeability of the gut epithelium. This increased gastrointestinal permeability is a key factor in producing symptoms of heatstroke by triggering the systemic inflammatory response, which lead to necrosis of organ tissues and multi-organ failure. Prevention of the gut permeability would offer a significant benefit in mitigation of the heat stress impact. Gut microbiota is critical in keeping mucosal barrier function. Stability of gut microbiota determines the ability of the organism to tolerate stress. Thus, modulating the intestinal microbiota by probiotics may be a novel, non-invasive therapeutic approach for prevention of heat stress-related adverse effects.We demonstrated that oral pretreatment of rats with Bacillus subtilis probiotic before exposure to heat protects animals from damage of intestine, translocation of bacteria from the gut, elevation of lipopolysaccharides and cytokines levels in the blood, increasing vesiculation of erythrocytes. We can speculate that some substances, produced by B. subtilis strain can contribute to the gut integrity and protection of the mucosal barrier function.
National Research Council, Italy
Title: Efficacy of readytoeat probiotic artichokes in modulating faecal microbial parameters in healthy subjects and patients with functional constipation
Time : 15:25 - 15:45
Paola Lavermicocca is a Biologist, Research Director and Leader of the Research Unit "Microbiology and quality of food productions". She is responsible for more than 20 scientific projects in the field of agro-food microbiology funded by European Commission/MIUR/Italian Regional Bodies/ Foundations/private funding bodies. Her research activities are focused on the microbiological aspects of food to improve their nutritional/functional quality and shelf life. She is an expert for European Commission, GSRT of Greece, Italian Ministry of Education, University and Research, Italian Ministry for Productive Activities and for Italian Regional Bodies. She is an author of about 140 publications (most ISI ranked) and has patent ownership on the application of bacterial strains in the field of functional foods.
A new line of probiotic products has been developed as an appealing option to the milk-based products to allow the daily consumption of probiotics. To evaluate the effects of ready-to-eat artichokes containing the probiotic strain Lactobacillus paracasei LMGP22043 on fecal microbial composition, biochemical parameters and symptom profile, clinical trials on healthy subjects and patients suffering from functional constipation were performed. Preliminary results provided evidences on the ability of the ready-to-eat probiotic food (daily dose of about 1x1010 CFU live probiotic cells) in modulating microbial parameters in constipated subjects. The administration for 15 days of the probiotic product to healthy volunteers demonstrated that the L. paracasei strain transiently colonized the gut of 17/20 subjects (median 6.87 log CFU/g feces), determined a general reduction in Enterobacteriaceae, antagonized Escherichia coli and Clostridium spp. and increased the genetic diversity of lactic population (Valerio et al., 2011). As concerns the patient group, 20 constipated patients were studied. Symptom profile was investigated using the Bristol stool form chart and the Gastrointestinal Symptom Rating Scale (GSRS) questionnaire. After 15-day administration of probiotic ready-to-eat artichokes, satisfactory relief of symptoms was recorded. Besides, Bristol chart cluster scores were significantly higher and GSRS constipation scores significantly lower at the end of administration compared to baseline. As for SCFAs production, a significant increase in propionic acid concentration was found. Present results suggest that the association of the probiotic L. paracasei with a food carrier rich in fibers such as artichokes, may represent a new strategy for favoring a daily supply of probiotics and attracting more consumers to vegetable food fortified with probiotic strains
State University of Campinas, Brazil
Time : 15:45 - 16:05
Mary L S Queiroz has completed her PhD from the University of Manchester, England, and her postdoctoral studies from WEHI, Melbourne, Australia. She is Full professor, director of the Laboratory of Immunopharmacology, Medical Faculty, UNICAMP, Campinas, Brasil. She has published more than 80 papers in reputed journals and presented several pioneer and original results in the literature, starting with the focus of her studies on the modulation by medicinal plants of immunohematopoietic regulatory mechanisms, aiming to increase or restore the host's own defenses which can inhibit infectious and malignant processes.
Purpose: Investigation of modulating effects of Chlorella on the medullar and extramedullar hematopoiesis and cytokine production of obese mice. Methods: Interaction between stromal cells and hematopoietic progenitor cells by long-term bone marrow culture. Growth and differentiation of bone marrow and spleen progenitors (CFU-GM) were obtained by clonal culture and cytokines by ELISA. Results: In obese mice we observed reduced capacity of stromal cell layer to support CFU-GM, decreased numbers of total non-adherent stromal cells, increased levels of IL-1, IL-6, TNF-alfa, TGF-beta, reduced levels of IL-10, and extramedular hematopoiesis. This latter finding is pioneer in the literature and might be related to the accumulation of macrophage in adipose tissue, a common feature in human and experimental obesity, which is considered responsible for the majority of complications observed in this disease. CV treatment restored all these changes to normal values. Conclusion: Additional findings, already published, were also pioneer in literature showing that prevention by CV of high-fat diet-induced insulin resistance in obese mice is due to improvement in insulin signaling pathway by increasing phosphorylation levels of IR, IRS-1 and Akt and reducing phosphorylation levels of IRS-1ser307 . We also found that CV prevents high-fat diet-induced dyslipidemia by reducing triglyceride, cholesterol and free fatty acid levels. Altogether our findings suggest that prevention by CV of the deleterious effects induced by high-fat diet is a good indicator for its use as a prophylactic agent against obesity-related complications.
Nazarbayev University, Kazakhstan
Time : 16:05 - 16:30
Introduction: Currently, the functional food is recognized as an alternative to pharmacotherapy. The objective of research is a synbiotic product "NAR", which includes probiotics and prebiotics. Probiotics are presented by cultures of Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus acidophilus, Bifidobacteriumlongum, Bifidobacteriumbifidum. The product is also enriched with fish collagen.The goal of this work is to investigate the clinical efficacy of a new synbiotic. Method: Double-blind, randomised, placebo-controlled trial, including 180 adult (age 30-89) with symptomes of metabolic syndrome whowere allocated to placebo or synbiotic group. We enrolled 90 adult in placebo group and 90 in synbiotic group. The two groups were similar for demographic and clinical characteristics. All patients signed a consent letter. The main criteria for inclusion were: Blood pressure: 130/90 mmHg; Raised fastingplasma glucose (FPG): >100 mg/dL (5.6 mmol/L), or previously diagnosed type 2 diabetes; Dyslipidemia: triglycerides (TG): =1.695 mmol/L and high-density lipoprotein cholesterol (HDL-C) =0.9 mmol/L (male), =1.0 mmol/L (female); Central obesity:waist:hip ratio >0.90 (male); >0.85 (female), or body mass index >30 kg/m2. All patients underwent clinical and laboratory evaluation, including completeblood tests, glucose test, glycosylated hemoglobin, total cholesterol + triglycerides, cholesterol, LDL, HDL plasma, immunogram, co-program. All patients were interviewed, the questionnaire included 200 questions related to diet, lifestyle and health. Synbiotic were used by patients in dose of 200 grams twice a day. The duration of applying the symbiotic were 90 days. Results: In the study group the percentage of colds were significantly lower than in placebo group, the majority of patients reported an improvement in abdominal discomfort, reduction in serum total cholesterol and low-density lipoprotein, weight loss of 8-9 percent in different age groups, stabilized frequency and consistency of stool (judging by the Bristol scale). Conclusion: As a result was exhibited the efficacy of a new synbiotic in adult with metabolic syndrome.