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evaluation of probiotic properties of lactobacillus reuteri strain using animal model 2012 PDF

180 Pages·2012·8.36 MB·English
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Preview evaluation of probiotic properties of lactobacillus reuteri strain using animal model 2012

EVALUATION OF PROBIOTIC PROPERTIES OF LACTOBACILLUS REUTERI STRAIN USING ANIMAL MODEL THESIS SUBMITTED TO THE NATIONAL DAIRY RESEARCH INSTITUTE, KARNAL (DEEMED UNIVERSITY) IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF THE DEGREE OF MASTER OF TECHNOLOGY IN DAIRY MICROBIOLOGY BY KATKAMWAR SNEHAL GOPALRAO B.Tech.(Dairy Technology) DIVISION OF DAIRY MICROBIOLOGY NATIONAL DAIRY RESEARCH INSTITUTE (DEEMED UNIVERSITY) KARNAL - 132001 (HARYANA), INDIA 2012 Regn. No. 2011006 Dedicated To My Lovely Family and Guide DAIRY MICROBIOLOGY DIVISION NATIONAL DAIRY RESEARCH INSTITUTE (DEEMED UNIVERSITY) KARNAL-132001 (HARYANA), INDIA Dr. R.K. Malik Principal Scientist CERTIFICATE This is to certify that the thesis entitled, “EVALUATION OF PROBIOTIC PROPERTIES OF LACTOBACILLUS REUTERI STRAIN USING ANIMAL MODEL ” submitted by Miss Katkamwar Snehal Gopalrao towards the partial fulfilment of the award of the degree of Master of Technology (Dairy Microbiology) of the National Dairy Research Institute (Deemed University), Karnal (Haryana), India, is a bonafide research work carried out by her under my supervision, and no part of the thesis has been submitted for any other degree or diploma. Dated: Acknowledgements….. “We think of the effective teachers we have had over the years with a sense of recognition, but those who have touched our humanity we remember with a deep sense of gratitude’. This was much more than a blessing God bestowed upon me when I was obliged to work under the guidance of my major advisor Dr. R.K. Malik, Principal Scientist, Dairy Microbiology Division, NDRI, Karnal, who as a compass activated the magnets of curiosity, knowledge, and wisdom in me. Words fail to elucidate my profound sense of veneration and indebtness for him for the constant encouragement, sagacious guidance, and inspiration for making right decisions throughout the study. I gratefully acknowledge the valuable suggestions given from time to time, by the members of my advisory committee: Dr. Sunita Grover, Dr. S. K. Tomar, Dr. Latha Sabikhi, and Dr. Suman Kapila. Their critical evaluation along with prolific ideas and suggestions helped me in the successful execution of my work. I also offer my sincere thanks to Dr T. K. Mohanty, Dr A. K Puniya, Dr. Sandip Khokhar, Dr. Sachidanad De, Dr. R. P. Singh, Dr. Malhotra, Dr. Gyan Singh and other faculty members of the Division for their unconditional help without any reservation throughout the study. My sincere thanks are humbly submitted to Dr. A.K.Srivastava, Director, NDRI, Karnal and Dr. Rameshwar Singh, Head, Dairy Microbiology Division, for providing all the necessary facilities to carry out the research. Financial assistance from ICAR in the form of institute fellowship is gratefully acknowledged. A formal line of appreciation would hardly meet the end of justice to record my gratitude for Purti ji for his fatherly affection, blessings and the technical assistance. My thanks are also extended to Randhir ji and Pravin Bhaiya for constant help without any reservation Words fail to express my deepest sense of gratitude and genuine love to the pillars of my strength, Santosh sir, T. P. Sir, Diksha, Neha mam, Renuka didi, Satvinder sir, Preeti di, Garima di, Deepika mam, Geetika di, Neha Tanwar di, and Gurpreet mam for having stood by me for the past two years during my good and bad times. Their inevitable love, care and critical but positive friendly suggestions made me a better human being. Special thanks to all of my seniours and juniours. I am also thankful from the very core of my heart to my friend Bhawna, Vandana, Vaibhao, Prity di, Naren and Renuka di for their wonderful support, company, friendship and immense help in the final drafting of this piece of work. My thanks are also extended to Samridhi, Ritu di, Kesha, Prachi, Pranoti, Pritee, Bulbul di, Sneha for appreciating my achievements, enthusiasm and helping me in all possible ways. A special note of acknowledgement is presented with courtesy to my Classmates Vaibhao, Narendra, Shashank, Sukirti, Dewashish, Bhanu, Vandna, Vaishali, Upasana, Ayesha and Bhawna again for their utmost help, care and concern. I express my heartfelt appreciation and thanks to my best friends Ashish G, Vinu Dada, Sarang, Nilay, Nishant for their friendly help and required cooperation. I feel much indebted to technical staff of our Division R. P. Singh sir for availing me the instrument facility. Thanks are also due to staff of animal house for providing the required facilities. I honorably express my deepest gratefulness towards my parents and my whole family for their godly influence upon my life and their countless blessings. I gratefully acknowledge the concern, affection and understanding of my brothers, which can only be felt and not expressed in words. Last but not the least, my humble and absolute obeisance to Almighty God who has guided me through all good and bad times and also for blessing me abundantly. Thanks a lot!!! Dated: July, 2012 Place: Karnal ABSTRACT Probiotics are live organisms which when administered in adequate amount confers health benefit to host by maintaining gut homeostasis. Probiotics exerts their beneficial effects beyond gastrointestinal health, as they have been found to improve certain inflammatory metabolic disorders such as hypertension, cardiovascular diseases, diabetes etc. there by reducing the biological risk factors associated with these life-style diseases. In the present investigation, based on a previous in vitro study in our lab, a total number of 8 Lactobacillus reuteri isolates were selected and reconfirmed as L. reuteri on the basis of physiological, biochemical and molecular characterization. Most of the isolate precipitated sodium taurocholate and sodium tauroglycholate indicating good bile salt hydrolase activity. Molecular identification revelaed the presence of bsh gene in one isolate i.e. L. reuteri LR 6. LR 6 was also found to exhibit maximum hydrophobicity for xylene (48.83%) and n-octane (36.37%). All the isolates exhibited good spectrum of antimicrobial activity against selected pathogens and have moderate antioxidative activity. In in vitro cholesterol assimilation studies, LR 9 exhibited maximum cholesterol assimilation followed by LR 6. Hypocholesterolemic activity of LR 6 was studied in vivo using rats as an animal model. After 60 days of feeding, significant reduction in serum cholesterol level was noticed in probiotic fed group, with reduction in total cholesterol by 33.36 % (79.24mg/dl), enhancement of HDL cholesterol by 29.30% (46.33mg/dl), reduction in triglycerides level by 25.25% (59.24mg/dl), reduction in LDL cholesterol by 68% (21.06mg/dl), reduction of VLDL by 25.23% (11.85mg/dl) and reduction in Atherogenic index by 79.93% (0.45). during immunomodulatory studies feeding of LR 6 caused significant increase in IgA concentration on 5th day. Maximum decrease in S. typhi counts was noticed in group IV on 8th day in different organs like liver, spleen and large intestine. The overall investigation shows that LR 6 can be used to develop functional foods to control hypercholesterolemia. र र | र र र र | : र , र , र , र र र | र 8 र र | र र र र र र | र र र र | र र र र 6 | र र र और | र र र र र 9 र र र 6 र | र र र 6 | 60 , र र | 33.36 % , 29.30% , 68% , र 25.25% , 25.23% और 79.93% | र र र र र र 6 5 | 8 र , , और | र 6 र र | CONTENTS Page Chapter Title No. 1.0 INTRODUCTION 1-4 2.0 REVIEW OF LITERATURE 5-36 2.1 Cardiovascular Diseases (CVD) 5 2.2 Cholesterol and Human Health 6 2.3 Background on Probiotics 8 2.3.1 History of Probiotic Organism 8 2.3.2 Definition of probiotics 9 2.4 Selection of Probiotic Strain 11 2.5 Proposed Mechanism of Action of Probiotics 12 2.6 Lactobacilli as Probiotics 13 2.6.1 Lactobacilli – General Characteristics 14 2.7 Suitability of Lactobacilli as Probiotics: 16 Selection Criteria 2.7.1 Acid and Bile Salt Tolerance 16 2.7.2 Antimicrobial Action 17 2.7.3 Adherence Properties 19 2.7.4 BSH Activity of Probiotics 21 2.8 Anticholesterolemic Activity of Probiotic 22 Organisms 2.9 Lactobacillus reuteri as a Probiotics 26 2.9.1 Historical Background of L. reuteri 27 A Morphology 27 B Ecology 27 C Biochemistry 27 2.9.2 Probiotic Attributes of L. reuteri 28 2.9.3 L. reuteri: Safety Aspects 29 2.9.4 Applications of Lactobacillus reuteri 31 2.10 Immunomodulatory Activity of Lactobacillus 32 Reuteri 2.11 Hypocholesterolemic Activity of Lactobacillus 35 Reuteri 3. MATERIALS AND METHODS 3.1 Bacterial Cultures 37 3.2 Maintenance and Propagation of Cultures 38 3.3 Purity and Confirmation of Cultures 39 3.4 Selection of L. Reuteri Isolates 39 3.4.1 Identification and Characterization of 39 Lactobacilli 3.4.1.1 Microscopic examination 39 3.4.1.2 Catalase test 39 3.4.1.3 Physiological 40 characterization a Growth at 15 and 40 450C 3.4.1.4 Biochemical 40 characterization of lactobacilli a Gas from glucose 40 b Argininie hydrolysis 40

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Probiotics are live organisms which when administered in adequate amount significant reduction in serum cholesterol level was noticed in probiotic fed .. treatment of body ailments has been mentioned even in Biblical scriptures.
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