Molecular characterization and probiotic potential of Kimere lactobacilli:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Göttingen
Cuvillier
2009
|
| Ausgabe: | 1. Aufl. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | IX, 117 S. Ill., graph. Darst. |
| ISBN: | 9783869550565 |
Internformat
MARC
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Datensatz im Suchindex
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| adam_text | Titel: Molecular characterization and probiotic potential of Kimere lactobacilli
Autor: Njeru, Patrisio Njiru
Jahr: 2009
TABLE OF CONTENTS
TABLE OF CONTENTS
Dedication.....................................................................................................................................................ii
TABLE OF CONTENTS.............................................................................................................................iii
LIST OF ABBREVIATIONS......................................................................................................................vi
ABSTRACT...............................................................................................................................................viii
ZUSAMMENFASSUNG............................................................................................................................ix
CHAPTER 1.........................................................................................................................1
MOLECULAR METHODS IN PROBIOTIC RESEARCH........................................................................1
ABSTRACT..................................................................................................................................................1
1 INTRODUCTION............................................................................................................................1
2 GENETIC FINGER PRINTING METHODS..................................................................................3
3 POLYMERASE CHAIN REACTION (PCR)-BASED METHODS...............................................5
4 HYBRIDIZATION METHODS.......................................................................................................6
5 REFERENCES...............................................................................................................................11
Chapter II........................................................................................................................20
Microbiological quality and biodiversity of lactobacilli from Kimere, a spontaneously fermented pearl
millet dough from Mbeere, Kenya (East Africa)........................................................................................20
ABSTRACT................................................................................................................................................21
1 INTRODUCTION..........................................................................................................................22
1.1 Kimere Production.........................................................................................................................23
2.2 Microbiological Quality of Kimere................................................................................................26
2 MATERIALS AND METHODS....................................................................................................28
2.1 Kimere sampling and sample preparation......................................................................................28
2.2 pH determination in Kimere...........................................................................................................28
2.3 Microbiological analysis of Kimere...............................................................................................29
2.4 Isolation and biochemical characterization of lactobacilli.............................................................29
2.5 Molecular characterization of lactobacilli......................................................................................30
2.5.1 Amplified ribosomal DNA restriction analysis (ARDRA)..............................................................................30
2.5.2 Species-specific PCR for L. fermentum.........................................................................................................31
2.5.3 16SrDNA sequencing....................................................................................................................................31
2.5.4 Pulsed-Field Gel Electrophoresis (PFGE)....................................................................................................31
2.6 Data analysis..................................................................................................................................32
Thesis Njeru P.N 2009
3 RESULTS.......................................................................................................................................32
3.1 Microbiological quality of Kimere.................................................................................................32
3.2 Biochemical characterization of lactobacilli..................................................................................33
3.3 Species-Specific PCR identification..............................................................................................38
3.4 ARDRA of Kimere isolates and type strains..................................................................................38
3.5 Partial sequence of the 16S rDNA.................................................................................................40
3.6 PFGE profiles of Kimere isolates...................................................................................................42
4 DISCUSSION.................................................................................................................................43
5 ACKNOWLEDGEMENTS............................................................................................................47
6 REFERENCES...............................................................................................................................47
Chapter III......................................................................................................................53
Strain specific immunomodulation by Lactobacillus fermentum strains isolated from Kimere; an African
fermented pearl millet (Pennisetum glaucum) dough...............................................................................53
Abstract.......................................................................................................................................................54
1 INTRODUCTION..........................................................................................................................55
2 MATERIAL AND METHODS......................................................................................................58
2.1 Bacteria cultures.............................................................................................................................58
2.2 Resistance to bile salts...................................................................................................................59
2.3 Acid tolerance................................................................................................................................60
2.4 Testing immunomodulation...........................................................................................................60
2.4.1 PBMCs Isolation...........................................................................................................................................60
2.4.2 Co-incubation of PBMCs and bacteria.........................................................................................................61
2.4.3 Cytokines assay.............................................................................................................................................61
2.5 Statistics.........................................................................................................................................61
3 RESULTS.......................................................................................................................................62
3.1 Resistance to bile salts and low pH................................................................................................62
3.2 Cytokine production patterns.........................................................................................................64
4 DISCUSSIONS...............................................................................................................................67
5 ACKNOWLEDGEMENTS............................................................................................................70
6 REFERENCES...............................................................................................................................70
CHAPTER IV....................................................................................................................74
Suppression Subtractive Hybridization of Kimere Lactobaciilus strains....................................................74
Abstract............................................................................... 75
iv
TABLE OF CONTENTS
1 INTRODUCTION..........................................................................................................................76
2 MATERIALS AND METHODS....................................................................................................79
2.1 Bacterial strains..............................................................................................................................79
2.2 Isolation of genomic DNA.............................................................................................................79
2.3 Suppression subtractive hybridization (SSH)................................................................................80
2.4 Cloning into a TA vector................................................................................................................81
2.5 Plasmid DNA isolation and Recombinants screening....................................................................81
2.6 PCR screening of isolates...............................................................................................................81
2.7 Dot hybridization...........................................................................................................................82
2.8 DNA Sequencing and analysis.......................................................................................................82
3 RESULTS.......................................................................................................................................83
3.1 Construction of SSH library...........................................................................................................83
3.2 Sequence Protein homology and DNA matches............................................................................85
4 DISCUSSION.................................................................................................................................88
4.1 Relationship between Kl-Lbl SSH library and cell-surface structures.........................................89
4.1.1 Exopolysaccharides (EPS)............................................................................................................................89
4.1.2 Lipoteichoic and Wall teichoic acids.............................................................................................................90
4.1.3 Peptidogfycans..............................................................................................................................................90
4.2 Relationship between Kl-Lbl SSH library and Immune modulation...........................................91
5 ACKNOWLEDGEMENTS............................................................................................................93
6 REFERENCES...............................................................................................................................93
Chapter V......................................................................................................................101
1 GENERAL DISCUSSIONS AND APPLICATION OF RESULTS............................................101
1.1 Probiotics and Allergy..................................................................................................................103
1.2 Probiotics and Inflammatory Bowel Diseases (IBD)...................................................................104
1.3 Probiotics and Autoimmune Diseases..........................................................................................106
1.4 Superantigens and Thl/Th2 balance............................................................................................106
1.5 Conclusions and recommendations..............................................................................................107
2 REFERENCES.............................................................................................................................108
Acknowledgement....................................................................................................................................113
List of Tables............................................................................................................................................115
List of Figures...........................................................................................................................................116
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| spelling | Njeru, Patrisio Njiru 1975- Verfasser (DE-588)139457585 aut Molecular characterization and probiotic potential of Kimere lactobacilli vorgelegt von Patrisio Njiru Njeru 1. Aufl. Göttingen Cuvillier 2009 IX, 117 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Kiel, Univ., Diss., 2009 probiotics cabt lactobacillus cabt Kenya cabt genetics cabt characterization cabt Pennisetum glaucum cabt Perlhirse (DE-588)4209339-9 gnd rswk-swf Probiotikum (DE-588)4338112-1 gnd rswk-swf Isolierung Mikrobiologie (DE-588)4193882-3 gnd rswk-swf Lactobacillus (DE-588)4166370-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Perlhirse (DE-588)4209339-9 s Lactobacillus (DE-588)4166370-6 s Probiotikum (DE-588)4338112-1 s Isolierung Mikrobiologie (DE-588)4193882-3 s DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022272852&sequence=000004&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Njeru, Patrisio Njiru 1975- Molecular characterization and probiotic potential of Kimere lactobacilli probiotics cabt lactobacillus cabt Kenya cabt genetics cabt characterization cabt Pennisetum glaucum cabt Perlhirse (DE-588)4209339-9 gnd Probiotikum (DE-588)4338112-1 gnd Isolierung Mikrobiologie (DE-588)4193882-3 gnd Lactobacillus (DE-588)4166370-6 gnd |
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| title | Molecular characterization and probiotic potential of Kimere lactobacilli |
| title_auth | Molecular characterization and probiotic potential of Kimere lactobacilli |
| title_exact_search | Molecular characterization and probiotic potential of Kimere lactobacilli |
| title_full | Molecular characterization and probiotic potential of Kimere lactobacilli vorgelegt von Patrisio Njiru Njeru |
| title_fullStr | Molecular characterization and probiotic potential of Kimere lactobacilli vorgelegt von Patrisio Njiru Njeru |
| title_full_unstemmed | Molecular characterization and probiotic potential of Kimere lactobacilli vorgelegt von Patrisio Njiru Njeru |
| title_short | Molecular characterization and probiotic potential of Kimere lactobacilli |
| title_sort | molecular characterization and probiotic potential of kimere lactobacilli |
| topic | probiotics cabt lactobacillus cabt Kenya cabt genetics cabt characterization cabt Pennisetum glaucum cabt Perlhirse (DE-588)4209339-9 gnd Probiotikum (DE-588)4338112-1 gnd Isolierung Mikrobiologie (DE-588)4193882-3 gnd Lactobacillus (DE-588)4166370-6 gnd |
| topic_facet | probiotics lactobacillus Kenya genetics characterization Pennisetum glaucum Perlhirse Probiotikum Isolierung Mikrobiologie Lactobacillus Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022272852&sequence=000004&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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