Madridge Journal of Food Technology

ISSN: 2577-4182

International Probiotics and Antimicrobial Proteins Conference

November 6-8, 2017, Barcelona, Spain
Accepted Abstracts
DOI: 10.18689/2577-4182.a1.004

Effect of Maize-Based Corn Fibre Combined with Lactobacillus rhamnosus LGG and the Pilus-Deficient Derivative GG-PB12 on Fecal Microbiota, Immune Function and Metabolism in Healthy Elderly (Saimes Study)

Adele Costabile*, Triana Bergillos-Meca, Pia Rasinkangas, Katri Korpela, Willem M. de Vos and Glenn R Gibson

Health Sciences Research Centre, Life Sciences Department, University of Roehampton, UK

The aging process leads to a potential decline in immune function and adversely affects the gut microbiota. Dietary intervention is one approach to affect gut microbiota composition and improved functioning of the immune system. We investigated the effects of a probiotic Lactobacillus rhamnosus GG (LGG) and pilus-deficient L. rhamnosus GG-PB12 combined with PromitorTM Soluble Corn Fibre (SCF, a candidate prebiotic) on fecal microbiota, metabolism, immunity and blood lipids in healthy elderly persons. A prospective, double-blind, placebo controlled, randomized, single centered, cross-over study in 40 healthy elderly subjects (aged 60-80 y) was carried out. Volunteers were randomized to consume either probiotic and prebiotic as synbiotic, prebiotic or placebo (maltodextrin) during 3-weeks. Three-week wash-out periods separated all the treatments. We assessed effects upon blood lipids, glucose, cytokines, NK cell activity, phenotype and intestinal microbiota composition. SCF decreased IL-6, which was not observed with the synbiotics. Consumption of L. rhamnosus GG combined with SCF increased NK cell activity compared to baseline in females and the older group. In the fecal microbiota analyses, the strongest community shifts were due to L. rhamnosus GG combined with SCF and SCF treatments. L. rhamnosus GG combined with SCF and L. rhamnosus GG-PB12 combined with SCF significantly increased the genus Parabacteroides. L. rhamnosus GG combined with SCF and SCF increased concentrations of Ruminococcaceae Incertae sedis. Oscillospira and Desulfovibrio slightly decreased in the L. rhamnosus GG combined with SCF group, whereas Desulfovibrio decreased also in the L. rhamnosus GG-PB12 combined with SCF group. L. rhamnosus GG combined with SCF reduced total cholesterol and LDL-cholesterol in volunteers with initially elevated concentrations. CRP decreased during L. rhamnosus GG-PB12 combined with SCF intervention compared to baseline.

Conclusions: Piliated L. rhamnosus GG was more potent at inducing beneficial effects compared to L. rhamnosus GG-PB12 by modulating the microbiome, increasing NK cell activity compared to SCF alone in older volunteers and decrease of the proinflammatory cytokine IL-6.

Dr. Costabile joined the University of Roehampton, Health Sciences Research Centre in August 2015 as a Lecturer in Nutrition. Dr Costabile research interest and publications focus on the interactions between human nutrition and gut microbiology. Her publication record has been built through ten years of research fellowship in Professor Glenn Gibson group at the University of Reading where she has lead several human feeding studies, often built on data from initial in vitro microbiology experiments which have resulted in a number of functional food products becoming established within the European market. Dr Costabile research outcomes have broadened our understanding of the interaction between gut microbiome and the host in both health and disease, as well as the modulation of gut microbiome through functional foods. Specifically, Dr Costabile has formed strategic alliances with leading researchers in the fields of human nutrition, metabonomics, lipidomics, immunology and food. Bio- processing. These collaborations have led to joint publications, grant applications and financial support.

Use of Probiotics to Control Gastrointestinal Nematodes in Sheep under Grazing Conditions, in Uruguay

America Mederos1*, Rafael Orihuela2, Alberto Bozzo3 and Georgget Banchero1

1Instituto Nacional de Investigación Agropecuaria, Uruguay
2CALSAL, Uruguay
3Private Sector, Uruguay

In Uruguay, anthelmintic resistance is a wide-spread phenomenon among sheep farms and there is a need to search for alternative strategies. The aim of this study was to evaluate non-chemical alternatives to control GIN in sheep under grazing conditions in Uruguay. Trial1 was conducted at the Research Station “La Estanzuela”, Colonia using 110 lambs distributed in five groups (n=22) with two repetitions each: G1= feed 16% protein concentrate (Control); G2=16% protein concentrate + 1% condensed tannins; G3= % protein concentrate + 4% condensed tannins; G4= 16% protein concentrate + condensed tannins extract as oral drench; G5= 16% protein concentrate + Saccharomyces cerevisiae CNCM I-1077 (LEVUCELL®SC).Trial2 was conducted at a commercial farm in Artigas using 90 lambs. Three experimental groups (n=30) were formed grazing native grasses: G1= feed 20% protein concentrate + Saccharomyces cerevisiae CNCM I-1077 (LEVUCELL®SC); G2= feed 20% protein concentrate (Control) and G3= feed 20% protein concentrate + 1% condensed tannins. Trial3 was similar to Trial 2 conducted during 2016.

The follow up period for Trial1 and Trial2 was from January- June 2015 and for Trial3 from January-June 2016. Production traits were recorded and fecal egg counts (FECs) were performed on individual animals. Infective larvae were recorded in animals and pasture.

Briefly, the main effect of probiotics treatment was a significant reduction in the number of treatments the animals required to thrive in Trial2 and Trial3 (31 vs 44 and 11 vs 30 respectively), in comparison with the control (p<0.05). In Trial1, FECs tended to be lower in G5 compared with G1 (2448 vs 2898) and G1 compared with G2in Trial2 (900 vs 1100).

Although probiotics have proven to be effective in controlling microbes, the results obtained from these exploratory trials are promising for further studies.

América Mederos has a Veterinary Medicine degree from the Veterinary School in Montevideo, Uruguay. During 1993 she completed her MSs at Reading University (UK) and his PhD at the University of Guelph in 2010 in Veterinary Epidemiology. Since 1991 she joined the National Research Institute for Agriculture and has been conducting research programs on Animal Health. Her main interest has been in the study of anthelmintic resistance and searching for alternative methods to control gastrointestinal nematodes in ruminants using non-chemical drugs such as bioactive forages and probiotics.

The Effects of Lactobacillus plantarum BSL and Lactobacillus rhamnosus R23 on the Blood Lipid Profile of Streptozotocin-Induced Diabetic Rats

Betty Sri Laksmi Jenie1*, Eko Farida2, Lilis Nuraida1 and Puspo Edi Giriwono1

1Department of Food Science and Technology, Bogor Agriculture University, Indonesia
2Department of Public Health, Semarang State University, Indonesia

Previous studies showed that intake of some lactic acid bacteria was able to inhibit the progression of diabetes mellitus by reducing blood glucose level and associated symptoms, for example, the lipid profile. The objective of this study was to investigate the effect of Lactobacillus plantarum BSL and Lactobacillus rhamnosus R23 isolated from different sources (sauerkraut and breast milk, respectively), on the blood lipid profile of diabetic rats. Diabetic rats were prepared by inducing rats using streptozotocin (40 mg/kg BB). The rats were divided into four groups (n=6), non-diabetic rats that received only normal diet (negative control), diabetic rats that received normal diet and phosphate buffer saline (PBS) as positive control and diabetic rats that received normal diet and L. plantarum BSL or L. rhamnosus R.23. During 30 day periods, the amount of food intake and the body weight gain of rats were measured. On day 30, all rats were sacrified and lipid profiles of rats were determined by CHOD-PAP enzymatic method. The results showed that administration of Lactobacillus plantarum BSL and Lactobacillus rhamnosus R23 decreased the total cholesterol and total trygliceride of ratsʼ serum. L. plantarum BSL increased HDL-c, but L. rhamnosus R23 decreased HDL-c. Administration of Lactobacillus plantarum BSL and Lactobacillus rhamnosus R23 also reduced the ratios of TC: HDL-c, TG: HDL-c, and LDL-c: HDL-c which are normally used as a predictor of cardiovascular diseases (CVD). These results suggest that Lactobacillus plantarum BSL and Lactobacillus rhamnosus R23 are potential as probiotic to improve blood lipid profiles in diabetic condition and developed as functional foods.

Keywords: Lactobacillus plantarum, Lactobacillus rhamnosus, diabetic rats, blood lipid profile

Lactobacillus plantarum Inhibits Mouse Colonic Inflammatory Response by Activating NLRP6

Chunfeng Wang*, Wentao Yang, Guilian Yang, Keyan Huang and Yanlong Jiang

Jilin Provincial Engineering Research Center of Animal Probiotics, Jilin Agricultural University, China

Many experimental with animal models, clinical trials, and cell culture studies they have been confirmed that Lactobacillus plantarum has the ability to inhibit the inflammatory response, however, its mechanism to inhibit the inflammatory response is unclear. In this study we used LPS to treat BMDM and then added Lactobacillus plantarum in it. The results suggest that Lactobacillus plantarum could promote the expression of NLRP6 in BMDM cells and form inflammatory complexes with Caspase-1 and ASC, which inhibit the secretion of extracellular interleukin (IL)-1β, and promote the secretion of extracellular IL-18. We also used siRNA to interfere with the expression of NLRP6 in BMDM, the result showed that promote the secretion of IL-1β and inhibit the secretion of IL-18. Importantly, pre-inoculation with Lactobacillus plantarum can effectively attenuate DSS-induced colitis in mice through the result of colonic scoring, colon pathological changes and weight loss indicators. In conclusion, Lactobacillus plantarum inhibits the inflammatory response by activating the Nod-like receptor (NLR) family member, NLRP6. This study has laid the foundation for the application of lactic acid bacteria in the treatment of colitis.

Professor Wang graduated from Jilin Agricultural University with masterʼs degree in Preventive Veterinary Medicine. Then she attended China Agricultural University to earn the Ph.D. degree. After the doctoral career in Beijing, she was admitted to Harvard University to pursue her post-doctoral research. During the last two decades, she has devoted herself in the research work for the development of novel animal micro ecological agents, construction of a mucosal delivery platform to express heterologous proteins utilizing engineering lactic acid bacteria which have been regarded as safe microorganisms, exploration of molecular immune mechanisms for the new functional lactic acid bacteria, preparations of environmentally friendly animal micro ecological formulations. All the initial innovative academic achievements have been published in the journals, such as Cell Transplantation, Virology, and Scientific Reports.

Characteristics of Lactic Acid Bacteria Isolated from Dadih Batu Bajanjang, Lembang Jaya, Solok District, West Sumatra

Endang Purwati1*, Hendri Purwanto1, Salam N Aritonang1, Fika Lindryani2 and Ferawati1

1Laboratory Biotechnology/ Technology of Product Husbandry, University of Andalas, Indonesia
2Students of Post Graduated Biotechnology Campus Limau Manis University of Andalas Indonesia

Dadih is a fermented product from buffalo milk using bamboo tubes. It is one of the innovative product that provides positive effects for human health with probiotics as requirements of fungsional food. The study aimed to characterize and determine the species of lactic acid bacteria isolated from dadih which has proven produced the best quality of probiotics located in Batu Bajanjang, Lembang Jaya, Solok District, West Sumatra, Indonesia. The research had been performed using the sample from different location (AN, AS, KI). This method identifying by macroscopic, microscopic and molecular using 16S rRNA technique. Furthermore, this research also determine nutritional content such as protein, fat, water content, pH and acidity. The result of nutritional content was not significant different for all sampel. The protein content are AN 5.58%, AS 6.08%, KI 6.68%, fat content are AN 6.4%, AS 7.0%, KI 7.2%, water content are AN 80%, AS 73%, KI 65%, pH are AN 4.14, AS 4.07, KI 4.02 and acidity are AN 1.35%, AS 1.71%, KI 2.12%, respectively. The number of lactic acid bacteria are AN 20.8x107CFU/g, AS 15.3x107CFU/g, KI 3.8x107CFU/g. However, the number of aerobic bacteria are AN 14.8x104CFU/g, AS 46.4x104CFU/g, KI 50x104CFU/g. The results showed the isolate is namely Lactobacillus fermentum strain NCC2970 with morphology bacil, gram-positive with catalase negative. It was potential a commercial starter to produce another fermented food.

Keywords: Dadih, fungsional food, lactic acid bacteria, 16S rRNA, probiotik

Prevalence of Bovine Trypanosomosis in and Around Nekemte Areas, East Wollega Zone, Ethiopia

Feyera Gemeda Dima* and Adem Abdellah

Jimma University, Ethiopia

A cross-sectional study was conducted from November 2010 to April 2011 to determine the prevalence of trypanosomosis and to identify the prevailing species of trypanosomes in cattle present in and around Nekemte. Blood sample were collected from ear vein of 400 cattle and then examined using thin and thick smear method followed by Buffy coat examination. Anemic status was determined by Packed Cell Volume (PCV). Out of 400 samples were examined, 36 (9%) were positive, out of which 26 (0.065%) had Trypanosoma vivax and 10 (0.025%) had T. congolense. The mean PCV of the infected animal is 19.36 and that of non-infected animal is 27.54, which indicates a significant difference between these animals. Trypanosomosis is a fatal and economically devastating disease and the major constraint to production by causing loss of the livestock. Therefore, the better strategies to prevent this disease includes: avoidance of animals from tsetse-infested areas, tsetse fly control by using different scientific methods (such as, sterile insect technique, use of accaricides), prophylactic use of trypanocidal drugs, keeping of trypanoresistant breeds and good husbandry practice for the prevention of the disease.

Keywords: Prevalence, Trypanosomosis, Bovine, East Wollega Zone, Nekemte Town, T. vivax, T. congolense.

Feyera Gemeda Dima is in Doctor of Veterinary Medicine in ground. He had done a research (DVM Thesis) on bovine Trypanosomosis in and around Nekemte areas.
He is working as a Lecturer for Jimma University with Five Years of work experience. He is a member of International Research organizations and also involved many publications.

Can Chronic Probiotic Intake Modulate Psychological Profile, Gut Microbiota and Body Composition of Women Affected by Normal Weight Obese Syndrome and Obesity? A Double Blind Randomized Clinical Trial

Giuseppe Merra2*, Antonino De Lorenzo1, Santino Gaudio1, Paola Gualtieri1, Silivia Barruco1, Massimiliano Marchetti3 and Laura Di Renzo1

1Department of Biomedicine and Prevention, University of Rome Tor Vergata, Italy
2Catholic University of Sacred Heart, Italy
3University Sapienza, Policlinico Umberto I, Italy

Background: Evidence of probiotics effects on gut function, brain activity and emotional behaviour were provided. Lactobacillus appears to reduce body fat mass, anxiety and dysphoria, and improve insulin sensitivity and glucose tolerance. Body dissatisfaction has been observed to be a risk factor for obesity and eating disorders. Probiotics can have dramatic effects on behavior through the microbiome-gut-brain axis, through vagus nerve. They produce a variety of neurochemicals, analogs of mammalian hormones involved in mood and behavior.

Aim: Given the link between gut microbiota, body composition, and the risk of psychiatric illness, we investigated whether chronic probiotic intake could modulate psychological state, eating behavior, and body composition of normal weight obese (NWO) and preobese-obese (PreOB/OB) compared to normal weight lean women (NWL).

Methods: 60 women were enrolled. Exclusion criteria included presence of intestinal bacterial overgrowth, history of chronic degenerative or infectious diseases, medication, smoke, drug or alcohol abuse. Subjects with acute diseases, severe liver, heart or kidney dysfunctions, endocrine disorders, cancer or other conditions capable of altering body composition were excluded. We categorized the subjects according to BMI, and % of total body fat (TBFat) into three groups: NWL, (BMI <25kg/m2, %TBFat<30); NWO (BMI <25 kg/m2, %TBFat ≥30); PreOB/OB, (BMI ≥25 kg/m2, %TBFat ≥30). At baseline and after 4 week of a probiotic oral suspension (POS) intake all subjects underwent to: anthropometric evaluation (body weight, height, waist and hip circumferences); body composition by dual X-ray absorptiometry (DXA); gut microbiota evaluation by Glucose Breath Hydrogen test (GHBT). Moreover, all patients were also assessed by means of self-report questionnaries (i.e. Eating Disorder Inventory, Beck Depression Inventory, State and Trait Anxiety inventory, and Body Uneasiness Test). POS contained Streptococcus thermophilus, Bifidobacterium animalissubspLactis, Streptococcus thermophiles, Lactobacillus bulgaricus, Lactococcus lactis subspLactis, Lactobacillus acidophilus, Lactobacillus Plantarum, Lactobacillus Reuteri (1.5 ×1010 colony-forming unit CFU for each/day). Multivariate of interest analyses, Pearson or Spearmanʼs rank correlation, and Partial Least Squares (PLS) were performed. Trial Registration: Id: NCT01890070

Results: Of the 60 women initially recruited, 8 did not meet inclusion criteria, 4 dropped out of the study voluntarily. We found a 22% of NWO, 26% of NWL, and 48% of PreOB/OB women. Significant differences (p<0.05) were highlighted between: NWL and NWO (TBFat, total body Lean, TBLean); NWO and PreOB/OB (Weight, BMI, TBFat, TBLean); NWL and PreOB/OB (Weight, BMI, TBFat, TBLean). After POS treatment, a reduction of TBFat (p<0.001) and syndrome of bacterial overgrowth (p<0.05), as well as lower psychopathological scores (i.e. depression, state anxiety, body dissatisfaction and bulimia) (p<0.05) were observed in NWO and PreOB/OB. BMI, and body composition variables co-varied with gut microbiota and psychological responses (p<0.001).

Conclusion: Four-week intake of selected probiotic, by modulating body composition, bacterial contamination, psychopathological scores and eating behavior of women affected by NWO syndrome and obesity, offers a tractable approach to problems related to obesity, psychological state and unhealthy eating. Further research is needed on a larger population and for longer period of treatment before definitive conclusions can be made.

The Effects of a Probiotic Protocol on Salivary Biomarkers, Heart Rate Variability, and Daily Wellness Scores in Elite Australian Athletes - Results of a Double-Blind Randomised Controlled Trial

Joanna Harnett1*, Andrew McKune2, Haydn Masters3 and Kate Pumpa2,3

1The University of Sydney, Australia
2University of Canberra Research Institute for Sport and Exercise, Australia
3Australian Rugby Union, Australia

Objective: To evaluate the effects of a probiotic protocol formulation on the salivary biomarkers sIgA, cortisol, alpha-amylase and psychometric measurements in elite athletes.

Methods: A double-blind randomised controlled trial was conducted over 17 weeks. Elite male Australian Rugby Union athletes were randomly assigned to receive either aprobiotic (n=11) or a placebo (n=10) supplement. The probiotic Ultrabiotic 60 or placebo was taken with food twice daily for 13weeks and SB Floractiv™ 250mg twice daily during athletes international travel period (9 weeks). The main outcome measured included salivary alpha-amylasesAA, cortisol, testosterone and secretory immunoglobulin A (sIgA), heart rate variability (HRV) and daily wellness scores including sleep quality, motivation and muscle soreness.

Results: There were no significant differences between the groups at baseline: age p=0.77, and weight (p=0.96). Multiple linear regression analysis identified alpha amylase and LF/HF ratio (HRV) were both independent predictors of general muscle pain and together account for 15.31% of the variance in general muscle pain in the placebo group (p < 0.0001). Sleep quality and motivation were both independent predictors of reducedmuscle pain and together account for 40.43% of the variance in general muscle pain (p< 0.0001). HRV and alpha-amylase were weakly associated with general muscle pain in the probiotic group.

Conclusion: The probiotic protocol trialed in this study resulted in a positive effect on sleep quality and motivation scores that is associated with a reduction in perceived muscle pain. Further research is required to evaluate this association.

Australian and New Zealand Clinical Trials Register: ACTRN12616000555459

Dr. Joanna Harnett holds a PhD in nutritional pharmacology and a bachelor and master degree in health science. She currently holds a fulltime teaching and research position within the Faculty of Pharmacy at The University of Sydney Australia. Her PhD (2013) explored the association between the intestinal microbiota and coeliac disease and included a randomized controlled trial evaluating the effects of a probiotic formulation on the composition of the faecal microbiota and symptom scores. Her current research activities include collaborating with Associate Professors Kate Pumpa (Sports Dietician) and Andrew McKune (Exercise Physiologist) exploring the role of probiotics in elite athletes.

Lactobacillus rhamnosus can interfere with Candida albicans Pathogenicity

Mariella Leaeo

Universidade de Taubaté – UNITAU, Brazil

Based on previous studies, which demonstrated that the consumption of probiotics was able to reduce the prevalence and amounts of Candida in the oral cavity of young and elderly individuals, we have been studied the effects of Lactobacillus rhamnosus on the pathogenicity of Candida albicans.

Aiming to investigate the influence of Lactobacillus rhamnosus intake on the development of candidiasis and cytokines release, in vivo experiments were performed. Candida suspensions were inoculated into the oral cavity of experimentally immunosuppressed mice for candidiasis induction. The animals were divided into experimental groups: candidiasis with no probiotic intake (F), candidiasis with probiotic intake during Candida inoculation (FP) and candidiasis with probiotic intake 14 days before inoculation with Candida (FPP); and control groups: (C), (CP) and (CPP) without inducing candidiasis with probiotic intake in the same manner as groups F, FP and FPP, respectively. After these periods, samples were collected from the oral cavity for yeast counts and, after euthanasia; the tongues of the animals were removed for histological analysis. Sera samples were also collected for analysis of IL-1 beta, TNF-alpha, INF-gamma, IL-12, IL-4, and IL-10. FP group showed lower Candida counts in the oral cavity, and the presence of Candida was almost not detected in FPP group. In tissues, the counts of fungi were significantly lower in FPP group, followed by FP. Groups that consumed probiotics also had lower histological and inflammatory infiltrates compared to F. Cytokines analysis demonstrated low concentrations of TNF-α, IL-12, IL-4 and IL-10 in all the groups, and no statistical difference between them. The production of IL- 6 could be better detected and the experimental groups that consumed the probiotic showed significant lower levels of this cytokine.

Other experiments investigated, in vitro, whether the interaction with Lactobacillus rhamnosus could interfere with the expression of virulence factors by Candida albicans. Therefore, these microorganisms were grown in biofilms, Candida strains were isolated and the expressions of the major virulence factors were investigated. The production of phospholipase, protease and hemolysin were observed in appropriate media; filamentation, after growth in serum and observation of germ tubes formation; biofilm formation, after growth in microtiter plates and reading in spectrophotometer; and hydrophobicity, by the use of xylene. The strains were also tested for antifungal sensitivity to amphotericin B, fluconazole and ketoconazole. The results were compared with strains of Candida grown in the absence of lactobacilli (control group). It was observed that Candida cells which interacted with L. rhamnosus (test group) showed significantly lower proteinase and hemolysin activity, when compared with control group. The germ tube formation and biofilm formation capacity also decreased significantly in tested groups, which also demonstrated alterations in susceptibility to antifungical drugs.

These results suggest that L. rhamnosus intake, especially preventively, may avoid or decrease the development of candidiasis, probably by interfering with the virulence factors of C. albicans and reducing its pathogenicity.

A Meta-Analysis to Establish the Effects of Probiotics on Growth Performance and Diarrhea Reduction in Swine

Marilen P. Balolong1*, Howell T. Ho2 and Dae-Kyung Kang3

1Department of Biology, University of the Philippines, Philippines
2Department of Biological Science and Biotechnology, Hannam University, Republic of Korea
3Department of Animal Resource Science, Republic of Korea

We conducted a meta-analysis to evaluate the effects of probioticson the growth performance and reduction of diarrhea in swine. We searched databases (e.g., PubMed, Scopus, and EMBase [ScienceDirect]) for papers written in English from 2005 to 2016. The inclusion criteria were as follows: studies using randomized and controlled experimental designswith pigs at any stage of development, with or without a pathogen challenge, published in peer-reviewed journals. Twenty six papers covering more than 800 experiments were considered for evaluation to establish the effects of probiotics on growth performance as measured by average daily gain (ADG), average daily feed intake (ADFI) and feed efficiency (FE). In addition, probiotic effects on diarrhea incidence (DI) and total coliform population (TC) and E. coli (EC) were also assessed.Probiotic supplementation of feeds increased ADG (mean difference = 23.4 g/day) and has improved feed efficiency (mean difference = −0.0188 kg feed/kg body weight). Although antibiotic supplementation contributed to more gain ADG (mean difference = 39.3 g/day), feed efficiency from administered probiotics was better (mean difference = 0.0313 kg feed/kg body weight). Mono-strain probiotics in feeds also favored growth performance ADG (mean difference = 21.4 g/day)and feed efficiency (mean difference = −0.0504 kg feed/kg body weight) than multi-strain probiotics with ADG (mean difference = 19.7 g/day) and (mean difference = −0.0037 kg feed/kg body weight), respectively,when added to feeds. Effects of probiotics on daily gain were more prominent in piglets but feed efficiency was more noticeable in adult pigs. Mono-strain probiotics were more effective in significantly reducing DIand TCs, whereas multi-strain probiotic preparations were effective in reducing EC. Probiotic supplementation also significantly reduced DI and TCs in adult pigs as well as EC in piglets. The magnitude of probiotic effects was similar to that of antibiotics in reducing DI when added to feed, regardless of the age of the pigs. Additionally, probiotics performed similarly to antibiotics in experiments using mono-strain and adult pigs to reduceTCs. We observed significant evidence of inter-experiment heterogeneity, which may have resulted from differences in study designs among experiments or from other factors not considered in our analysis. Overall, our results affirm the benefits of probiotic administration in swine for improved growth performance and reducing diarrhea.

Probiotics and/in P4: Personalization, Prevention, Prediction and Patients

Nadiya Boyko1*, Tamara Meleshko1, Viktoriia Bati1 and Olga Levchuk1, 2

1Uzhhorod National University, Ukraine
2Astra-Dia, Diagnostic Centre, Ukraine

New ideas about the uniqueness of the human microbiome and its decisive role in the health of the population today substantiate the idea of individualization of treatment approaches for patients (patient stratification) and individualized prevention. The latter being the prevention of diseases of the population through the specific implementation of personalized nutrition. Embodied in this is the selection of different biotics (pro-, pre-, syn-, pharma- and immuno-biotics).They need to be 1) unique to each individual; 2) specific, and therefore diagnostic for each type of disease.

Based on these approaches we created the new human pharmabiotics. At the core of both biotics modifiers is the initial composition of synergistically selected microorganisms of the normal commensal microbiota and prebiotic ingredients prescreened and extracted from edible plants. Their composition can be changed and adapted according to the individual patientsʼ requirements and to any nosology. This is done by selecting the necessary components and their specific combination in accordance with existing bioactive substances databases and their impact on representatives of commensal, pathogenic and opportunistically pathogenic microbiota.

We had tested their ability specifically modulate local and systemic immune response in experiments in numerous animal models and also on human dendritic cells derived from peripheral blood monocytes. One of the key aspects in the development of such specified, truly personalized bioticsis the geographical aspect, because on the one hand the microbiome state is adjusted by different local epigenetic factors including diet (composition, properties, content of biologically active compounds and impact) and on the other hand, determined by local factors.

In light of the aforementioned, we believe that such biotics are as promising as mono- or fixed structure pro-, pre- and synbiotics due to the fact that in addition to the impact of functionality regulation, their selectivity also helps to restore the individualsʼ microbiome biodiversity.

Nadiya Boyko, Ph.D., DSc. in Microbiology, 5 years sabbatical Research Fellow in Laboratory of Mucosal Immunology, University of Pennsylvania, USA, now Professor at the Faculty of Medicine of Uzhhorod National University, Director of the R&D Centre of Molecular Microbiology and Mucosal Immunology. She has published more than 200 papers, including chapter in Mucosal immunology Elsevier press. She is an expert is pharmabiotics and personalised nutrition, regulation of human [gut] microbiota for prevention of non communicable diseases. She is member of SMI, SOMED, ASA, SMU.
International project: FP7 (BaSeFood, JSO-ERA), CEE: CAPINFOOD, FoodWARD (Erasmus+), COST: BacFoodNet, ODiNand SKIN

Eubiotics: Future in Biotechnology Industry

Naheed Mojgani

Biotechnology Department, Razi vaccine and Serum Research Institute-Agriculture Research Education and Extension Organization & Research & Development Department, IR Iran

With the ban on the use of antibiotics as growth promoters and increasing consumer awareness with respect to preventive health care and nutrition, high demand for safe human food has fueled eubiotic market. The term Eubiotics is mainly used in the feed industry where it refers to a healthy balance of the micro-flora in the gastrointestinal tract. Bacteritic preparations intended for correction of biocenosis of the mucous membranes may be referred to as eubiotics. Poultry feed dominates the eubiotic market owing mainly to the large number of population consuming poultry meat in the world. The demand for eubiotics in animal feed is projected to grow during the forecast period, owing to the rapidly increasing global demand for quality animal products. Probiotics, prebiotics, essential oils and organic acids are often referred to as eubiotics. In this context, Probiotics account for the major share in poultry feed. It is hypothesized that different eubiotic combinations would have a synergistic effect in improving animal performance.

In Iran, in last decade a number of local probiotic manufacturers have evolved which are producing high quality probiotic feed additives for animals. While, this trend is still in its developing and pillaring stages, a number of researches on eubiotic products with medical and animal applications are also on its rise. According to global estimates, Europe dominates the eubiotics market followed by North America, while, China, India, and Brazil are the developing countries where eubiotic market is also growing.

The biotechnology industry is now creating an opportunity for growth in the eubiotics sector. A number of eubiotic drugs are being developed especially for treating dysbiosis. However, factors such as low shelf life of eubiotics and high cost of eubiotics leading to the rise in cost of production are restraining the growth of the market. With high tech approaches and biotechnological innovations we foresee a bright future for eubiotic products in Iran. Hopefully with entering the new era of Biotechnology we will soon witness highlight eubiotic products in the country with wide applications.

Dr. Mojgani is an Associate professor at Razi Vaccine and Serum Research Institute has vast experience in the field of Medical Microbiology, Molecular Biology and Vaccinology. Her main area of research is on Probiotics, antibacterial peptides, designing and formulation of probiotic products, bacterial conjugate vaccines, tuberculin production, nano-encapsulation etc. She has successfully contributed in the production of Probiotic supplements for animals and humans and transferred the know-how to local manufacturers in the country. Her continuous efforts, dedication and hard work in the field of Probiotic have led her to be recognized in the country as an eminent specialist in the field.

Inhibition of Salmonellosis with Produced Conjugated Linoleic Acid by Linoleate Isomerase of Rumen Fungus

Rabar M. Abdulrahman

Department of applied Science, Kahramanmaras Sutucu Imam University, Turkey

During the last two decades, the epidemiology of food borne pathogens has changed rapidly along with the alterations in the social atmosphere and the ability of pathogens to adapt to new niches. Food is a source of transmitting diseases through which more than 200 diseases are transmitted. Salmonella is the most commonly suggested cause of food borne disease which constitutes a major public health problem in many countries. In this study, we have isolated anaerobic fungi (Orpinomyces sp. and Neocallimastix sp.) from rumens feces. Then by growing them within medium containing only Linoleic Acid as a source of energy, we have identified those that are capable of converting Linoleic acid into Conjugated linoleic acids. The produces CLA (conjugated linoleic acid) has been used as antibacterial to show their effects of Salmonellosis. The result of this study showed that the produced CLA has antibacterial effect on all four strains of Salmonella sp. (Salmonella enterica, Salmonella Typhi ATCC 14081, Salmonella sp.1 and Salmonlla sp.2) used within this study.

Keywords: Salmonellosis, Linoleic Acid, Conjugated linoleic acid, antibacterial.

Rabar M. Abdulrahman is a PhD Student. He completed his Masters in Medical Microbiology from the University of Ulster. He then worked at the Medical Microbiology Department, Koya University, and served as Assistant Lecturer at the University in Iraq.

Safety Evaluation and Antimicrobial Properties of Lactobacillus pentosus 22C Isolated from Iranian Traditional Food

Saeed Mirdamadi*, PariaMotahari and Mehran Kianirad

Department of Biotechnology, Iranian Research Organization For Science and Technology (IROST), Iran

Today, safety is a priority in food and dairy industry and an important step for introducing the traditional products for industrial production.

Determination of antimicrobialactivity and evaluation of the safety of candidatestrains based on antibiotics susceptibility and virulence potential genes are the most important safety assessment steps. The ability to inhibit other bacteria may allow the probioticto inhabit a niche; and increase its ability to competitivelyinhibit other gastrointestinal microbes and pathogenicbacteria. Moreover, antimicrobial activity may control fermentation and increase the shelf life of foodproducts. Purified antimicrobial compounds can be used as additives, but In situ production of antimicrobialcompounds using a producer starter culture is regarded as amore commercially attractive strategy in fermented food.

This study aimed toevaluate the safety and antimicrobial potential of Lactobacillus pentosus22C, isolated from traditional yogurt fromKermanshah province, Iran. Strain 22C showed no undesirableamino acid decarboxylase and β-hemolytic activities. The isolate was assessed for the incidence of virulence genes (gelE, efaAfm, efaAfs, ace, espfs, cylM, cylA and cylB), sensitivity to various antibiotics and virulence phenotypes. The strain produced an antimicrobial molecule named pentocin22C, a small peptide with a relative mass between 5 and 10 kDa. Bacterial inhibition was pH-independent, with greater activity at pH 4-6. Purified or semipurified antimicrobialagents can be used as bio-preservatives, and theproducing strain can be used as a Bacteriocin producing starter culture to improve food safety.

Probiotics Reduce the Risk of Antibiotic-Associated Diarrhea in Adults (18–64 Years) but not the Elderly (>65 Years): A Meta-Analysis

Sadegh Jafarnejad1*, Sakineh Shab-Bidar2, John R. Speakman3, Karim Parastui4, Milad Daneshi-Maskooni2 and Kurosh Djafarian1

1Department of Clinical Nutrition, Tehran University of Medical Sciences, Iran
2Department of Community Nutrition, Tehran University of Medical Sciences, Iran
3Institute of Biological and Environmental Sciences, University of Aberdeen, Scotland
4Department of Cellular and Molecular Nutrition, Tehran University of Medical Sciences, Iran

Background: Antibiotic-Associated Diarrhea (AAD) is a common problem in adults and elderly patients due to the widespread use of antibiotics in this population. Multiple previous systematic reviews have demonstrated an association between specific probiotics and decrease of AAD, especially in children. As there is no specific analysis concerning the elderly patients, we decided to focus on adults, especially elderly people.

Methods: We performed a systematic review of the literature regarding the use of probiotics in the treatment of AAD in adults (18–64 years old) and elderly subjects (≥65 years old). We identified 436 articles that met the search criteria. Thirty randomized controlled trials met the predefined inclusion criteria and were included in the meta-analysis.

Results: There was considerable heterogeneity among the trials (p < .001); thus, subgroup analyses were performed. The metaanalysis resulted in a pooled relative risk (RR) of AAD of 0.69 (95% confidence interval [95% CI]: 0.62–0.76) in a fixed effects model and 0.58 (95% CI: 0.48–0.71) in a random effects model, as compared with placebo. The positive association between intake of probiotic and reduced risk of AAD was observed in adults (RR, 0.47; 95% CI: 0.4–0.56). In contrast, in elderly patients, there was no positive effect (RR, 0.94; 95% CI: 0.76–1.15) of probiotic use and AAD.

Conclusion: In summary, the results emerging from our meta-analysis suggested that adjunct probiotic administration is associated with a reduced risk of AAD in adults but not in elderly people.

Keywords: Probiotics; Antibiotic-Associated Diarrhea; Diarrhea; Elderly; Adults; Aged

Developing a Vaccine against the Porcine Rotavirus using Lactobacillus Plantaram Cells

Seria Shonyela

Jilin Agricultural University, China

Porcine rotavirus infection is a significant cause of morbidity and mortality in the pig industry necessitating the development of effective vaccines for the prevention of infection. Gut mucosal immune responses are likely to play significant role in protective immunity against rotavirus infection because rotaviruses are enteric pathogens.

Lactic acid bacteria (LAB) are Gram-positive, nonpathogenic microorganisms that are gaining to a great concern as antigen producers for improvement of live vaccine vectors. Heterologous proteins of dissimilar source have been effectively expressed in various LAB species. Recombinant L. Plantaram NC8 strains have been shown to induce specific local and systemic immune responses against a range of antigens.

The objective of this study was constructing a Lactobacillus Plantaram NC8 strain expressing the heterologous VP7 porcine rotavirus protein and investigating its outcome and its ability acting as an antigen delivery system for oral vaccinations on mice.

The expression of recombinant pSIP409-VP7-DCpep was confirmed by SDS-PAGE and Western blot analysis and surface-displayed expression on L. Plantarum was verified by immunofluorescence. Mice orally immunized with recombinant protein-expressing L. Plantarum produced high levels of serum immunoglobulin G (IgG) and mucosal IgA. The IgA titters from mice immunized with NC8-pSIP409-VP7-DCpep were higher than titters from pSIP409-VP7-DCpep -immunized mice. The induced antibodies demonstrated neutralizing effects on RV infection.

The outcome of this study is a hopeful step toward developing a vaccine against the porcine rotavirus using Lactobacillus plantaram cells as bioreactors for competent antigen production and delivery to the mucosal surface.

In vitro and In vivo Safety Analysis of Enterococcus facieum 2C Isolated from Breast Milk

Soodabeh Khalkhali1,2* and Naheed Mojgani3,4

1Department of Microbiology, Islamic Azad University, Shiraz, Iran
2Department of Microbiology, Islamic Azad University, Iran
3Biotechnology Department, Razi vaccine and Serum Research Institute, Iran
4Research & Development Department, IR Iran

Safety analysis of the probiotic bacteria is an obligatory characteristic evaluated prior to introducing them to food or pharmacological industry. This study was designed to evaluate in vitro and in vivo safety of Enterococcus faecium, a probiotic candidate isolated from human breast milk.

E.faecium isolated from breast milk was studied for its hemolytic activities, phenotypic antibiotics resistance profile and antibacterial activities in in vitro condition. During in vivo investigations, the oral toxicity of the mentioned probiotic strain was evaluated in Wistar Male rats. The animals were fed daily with dose of 1 x 1011 CFU/kg of body weight, respectively, for 21 consecutive days and their hematological, biochemical parameters, organ weight, body weight and common health features were recorded.

The results revealed that E.faecium 2C was non-hemolytic, sensitive to majority of tested antibiotics and was able to inhibit the growth of several pathogenic bacteria. During in vivo investigations, the Wistar male rats fed orally fed rats survived during the test period and showed normal growth and development. There were no adverse effects on the general condition, behavior, growth, feed and water consumption, hematology, clinical chemistry values, organ weights and histopathologic analysis of the rats. Results of this study demonstrate that consumption of strain E. faecium2C, even in large quantities, is not associated with any obvious signs of toxicity in Wistar rats. All physiological and biological health parameters including body and organ weights, biochemical blood and serum analysis revealed the safety of E. facieum isolate in study. None of the vital organs showed the sign of bacteremia or infectivity in the tested rat models.

E.faecium strain isolated from human breast milk is a safe probiotic with several beneficial properties; hence, it can be introduced for use as supplement for man and animals.

Ms. Soodabeh Khalkhali, born on January 2, 1981 in Tehran Iran, is a Ph.D. candidate (Microbiology) at Islamic Azad University Shiraz Iran. Her masters and PhD thesis has been on Probiotic bacteria and has published couple of papers on the importance of enterococcus species and significance of candidate probionts in mothersʼ milk. Ms. Khalkhali is also a part time teacher at a university in Tehran.

Purification and Characterization of Β-Galactosidase from Probiotic Pediococcus acidilactici and its use in Milk Lactose Hydrolysis and Galactooligosaccharide Synthesis

Suman Singh*, Preeti Chanalia, Dimpi Gandhi and Pooja Attri

Department of Biochemistry, Kurukshetra University, India

β- galactosidase is a commercially important enzyme that was purified from probiotic Pediococcus acidilactici. The enzyme was extracted from cells using sonication and subsequently purified using ammonium sulphate fractionation and successive chromatographies on Sephadex G-100 and Q-Sepharose. The enzyme was purified up to electrophoretic homogeneity by 3.06 fold with specific activity of 0.883 U/mg and yield of 28.26%. Molecular mass of β-galactosidase estimated by SDS-PAGE and MALDI-TOF was 39.07 kDa. The enzyme is a heterodimer with subunit mass of 15.55 and 19.58 kDa. The purified enzyme was optimally active at pH 6.0 and stable with more than 97% activity in a pH range of 5.8-7.0. Purified β-galactosidase was optimally active at 50°C. Kinetic parameters Km and Vmax for purified enzyme were 400µM and 1.22×10-1 U respectively. Its inactivation by PMSF confirmed the presence of serine at the active site. The metal ions had different effects on enzyme. Ca2+, Mg2+ and Mn2+ slightly activated the enzyme whereas NH4+, Co2+ and Fe3+ slightly decreased the enzyme activity. Determination of thermodynamic parameters revealed that the thermostability of β-galactosidase is less at higher temperature (60°C). Purified enzyme effectively hydrolysed milk lactose with lactose hydrolysing rate of 0.047 min-1 and t1/2 of 14.74 min. This is better than other studied β-galactosidases. Both sonicated Pediococcus acidilactici cells and purified β-galactosidase synthesized galactooligosaccharides (GOSs) as studied by TLC at 30% and 50% of lactose concentration at 47.5°C. These findings indicate the use of β-galactosidase from probiotic bacteria for producing delactosed milk for lactose intolerant population and prebiotic synthesis. pH and temperature optima and its activation by Ca2+ shows that it is suitable for milk processing.

Dr. Suman Singh is presently working as an Associate Professor, Dept of Biochemistry Kurukshetra University, Kurukshetra. She did her masterʼs from National Dairy Research Institute, Karnal (India) and subsequently did Ph.D. in Biochemistry from Kurukshetra University, Kurukshetra. For the last 14 years, she is teaching at post graduate level and guiding research. She has also worked at the German Institute of Human Nutrition, Potsdam-Rehbrucke, Germany. She has completed two research projects and currently running a Young Scientist Award cum research project funded by Department of Science and Technology, New Delhi India. She has published more than 30 research papers in accredited scientific Journals. She is working on strain characterization Pediococcus species after exploring its in-vitro probiotic potential. Purified enzymes from the strain under study are being studied to be used in dairy, food and meat industries. Her focus is to find out action molecules and biomarkers of probiotics.

Oligosaccharides Released from Milk Glycoproteins are Selective Growth Substrates for Infant-Associated Bifidobacteria

Sercan Karav

Canakkale Onsekiz Mart University, Turkey

Milk, in addition to nourishing the neonate, provides a range of complex glycans whose construction ensures a specific enrichment of key members of the gut microbiota in the nursing infant, a consortium known as the milk-oriented microbiome. Milk glycoproteins are thought to function similarly, as specific growth substrates for bifidobacteria common to the breast fed infant gut. Recently, a cell wall-associated endo-β-N-acetylglucosaminidase (EndoBI-1) found in various infant-borne bifidobacteria was shown to remove a range of intact N-linked glycans. We hypothesized that these released oligosaccharide structures can serve as a sole source for the selective growth of bifidobacteria. Here, EndoBI-1 was used to release these N-glycans from concentrated bovine colostrum at the pilot scale. EndoBI-1-released N-glycans supported the rapid growth of Bifidobacterium longum subsp. infantis, a species that grows well on human milk oligosaccharides, but did not support growth of Bifidobacteriumanimalis subsp. lactis, a species which does not. Conversely Bifidobacterium longum subsp. infantis ATCC 15697 did not grow on the deglycosylated milk protein fraction clearly demonstrating that the glycan portion of milk glycoproteins provides the key substrate for growth. Mass spectrometry-based profiling revealed that B.longum subsp. infantis consumed 73% of neutral and 92% of sialylated N-glycans, while B.animalis subsp. lactis only degraded 11% of neutral and virtually no (<1%) sialylated N-glycans. These results provide mechanistic support that N-linked glycoproteins from milk serve as selective substrates for the enrichment of infant-borne bifidobacteria capable of carrying out the initial deglycosylation. Moreover, released N-glycans are better growth substrates than the intact milk glycoproteins suggesting that EndoBI-1 cleavage is a key initial step in consumption of glycoproteins. Finally, the variety of N-glycans released from bovine milk glycoproteins suggests they may serve as novel prebiotic substrates with selective properties similar to those of human milk oligosaccharides.

Probiotic Lectin Systems Potential: The Glycoconjugate Binding Strategies

Lakhtin V.M., Lakhtin M.V., Afanasiev S.S. and Aleshkin V.A.

G.N.Gabrichevsky Research Institute for Epidemiology and Microbiology, Moscow, Russia

Innate protection involving lectin systems is of increasing attention of investigators. The human protection using glycoconjugates (GC) recognizing systems is important immune ingredient against infectious and autoimmune diseases. Probiotic lectin systems (PLS) possess multiple directions of actions useful for human. Aim: To propose the GC-strategies for evaluation the strain potential to select strain ingredients and construct probiotics. Materials and methods: Probiotic bacteria were from collection of G.N.Gabrichevsky Research Institute for Epidemiology and Microbiology. PLS were identified and isolated using isoelectrofocusing in polyacrylamide gel. They were identified on blots using GC-biotin ( and streptavidin-peroxidase in the presence chemiluminescent substrate in a real time in BioChemi System (UVP). Proteins were localized with SYPRO protein blot stain (Bio-Rad). Results and discussion: Different GC-binding PLS (> 27 kD, pI 4-8) were identified and isolated. PLS were localised as mosaics within protein massifs. The use of GC ( creates basis for constructing metabiotic and metabolomebiotic therapeutics of system action. The following activities of strain metabolites can be predicted: antipathogenic activities (the use of GC imitating microbial cell surface and wall structures: peptidoglycans, sialylated glycans, glycoantigens, others); prebiotic/ synbiotic actions (GC imitating prebiotic structures: derivatives of L-fucans; modified D-galactoside oligomers, etc.); increasing human protection (GC regulating peritoneal macrophages, complement system lectin components: (phospho) mannans and other polysaccharides, glycoantigens); on-duty support against cancerogenesis (normal cell surface décor panels of GC delivered); on-duty support of immunomodulation (panels of GC as antigens: peptidoglycans, others). PLS effectively imitated antimicrobial activities of bifidobacterial and lactobacillar cell probiotics. Conclusions: Results indicate the prospects of PLS as the new important functional metabolite ingredients of probiotics to use for prophylaxis and therapy of infectious and non-infectious diseases. The choice of fine specific (PLS—GC)-type interactions needed can be extended using a broad panel of GC.

Vladimir Lakhtin has his expertise in study of glycoproteins, lectins and their complexes, multistrain probiotics, antimicrobials, and innate immunity (complement system, etc.) upon infectious and non-infectious diseases. His functional evaluation models based on glycoconjugate recognition and responses of human biotope microbiocenoses and innate protective systems creates new perspective possibilities in diagnostics and prognostics in the field of medical biotechnology and clinical microbiology

Screening of Purine Nucleosides Degrading Probiotic Lactic Acid Bacteria as a Novel Approach in Management of Hyperuricemia

Neha Pandey and Shilpa Vij

National Dairy Research Institute, Karnal, Haryana, 132001

Out of 20 different curd and fecal samples, 76 Gram positive and catalase negative rods were isolated and screened for purine nucleoside degradation ability. A HPLC device equipped with a Photo diode array (PDA) detector was used to evaluate the ability of these isolates to degrade inosine and guanosine, the two key intermediates in purine metabolism by incubating these isolates with inosine-guanosine solution (1.25mM) for 2 h at 37o C. Contents of remaining inosine and guanosine were identified at 254 nm by retention time of 7.48 and 6.46 min, respectively, and quantified by interpolation of calibration curves. To evaluate the degradation of purine compounds by cell-free extracts of LAB, the cells were sonicated for 30ʼX3 (pulse rate- 5sec on/ 5 sec off) and activity evaluated through HPLC. Eleven isolates viz. D3A, D1AB, D1F, D3E, D6F, D2A, D5F, D3D, D3B, D1AC, D1AA were screened as positive for purine nucleoside degradation ability. These organisms were then evaluated for their potential probiotic attributes. Out of these 11 isolates only six isolates viz. D3A, D1AB, D2A, D6F, D5F and D1AA were found to be most acid tolerant and selected for further in vitro evaluation. When tested for bile tolerance only three isolates viz. D6f, D5F, D1AA and D3A were found to resist 2% bile and showed highest viability among the tested six strains. Further evaluation of the isolate for antimicrobial activity showed that the all the screened four isolates showed a good antimicrobial activity against Gram positive bacteria viz. Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microccus luteus and other Lactobacillus species. Our work shows that our isolates apart from purine degradation are good probiotics as proved by their acid and bile tolerance. Also, the antimicrobial activity showed by the isolate can give an added advantage to the isolate to compete and survive in the GI tract.

Dr. Neha Pandey is working as DST Women Scientist-A in Dairy Microbiology (DM) Division in National Dairy Research Institute (NDRI), Karnal, Haryana, India since 24th June 2016 under the mentorship of Dr. Shilpa Vij (Principal Scientist, NDRI, Karnal). She has completed her Ph.D in Dairy Microbiology from NDRI Karnal in 2013 and has eight publications in journals of national and international repute. For carrying out present study the authors highly acknowledge the funds received from Department of Science and Technology (DST), India.