Protocols in Actinobacterial Research: Springer Protocols Handbooks
Autor Ramasamy Balagurunathan, Manikkam Radhakrishnan, Thangavel Shanmugasundaram, Venugopal Gopikrishnan, Joseph Jerrineen Limba Engleză Paperback – 12 iul 2021
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Specificații
ISBN-13: 9781071607305
ISBN-10: 1071607308
Ilustrații: XXXI, 225 p. 27 illus.
Dimensiuni: 178 x 254 mm
Greutate: 0.46 kg
Ediția:1st ed. 2020
Editura: Springer Us
Colecția Springer
Seria Springer Protocols Handbooks
Locul publicării:New York, NY, United States
ISBN-10: 1071607308
Ilustrații: XXXI, 225 p. 27 illus.
Dimensiuni: 178 x 254 mm
Greutate: 0.46 kg
Ediția:1st ed. 2020
Editura: Springer Us
Colecția Springer
Seria Springer Protocols Handbooks
Locul publicării:New York, NY, United States
Cuprins
I. Sample collection and isolation of actinobacteria
o Collection and transportation of samples
o Soil
o Sediment
o Water
o Plants
o Insects
o Fisheso Animals
o Earthworms
o Processing and pretreatment of samples
o Soil
o Sediment
o Water
o Plants
o Animals
o Insects
o Earthworms
o Fishes
o Sample pre-treatment for the isolation of actinobacteria
o Physical treatment
o Chemical treatment
o Biological treatment
o Isolation and enumeration of actinobacteria -o Standard spread plate method
o Recover and purification of actinobacterial cultures
II. Dereplication and ex situ conservation of actinobacteria
o Dereplication
o Colony morphology
o Micromorphologyo FT-IR analysis
o RFLP analysis
o Ex situ conservation
o Slant stock
o Glycerol stock
o Soil stock
o Lyophilisation
o Immobilized beadsIII. Characterization and identification of actinobacteria
o Phenotypic characterization
o Microscopic
o Cultural
o Physiological
o Chemotaxonomic analysis
o Cell wall amino acids
o Cell wall sugars
o Cell wall lipids
o FAME analysis
o Menaquinone analysis
o Molecular characterization
o DNA isolation
o PCR Amplification of 16s rRNA gene
o Sequencing
o BLAST analysis
o GenBank Submission
o Phylogenetic tree construction
IV. Metagenomic analysis of uncultured actinobacterial diversity
o Sample collection
o DNA isolationo Amplification
o Clone library construction
o Sequencing
o Metagenomic analysis
o GenBank submission
V. Screening of actinobacteria for biological activities
o Screening for antimicrobial activity
o Cross streak method
o Agar plug method
o Well diffusion
o Disc diffusion
o Broth dilution method - MTP Assay
o MABA
o Screening for antiinfective or anti-inflammatory activity
o Screening for anti TB activity
o Agar dilution
o LRP assay
o Broth dilution method
o Bactec MGIT 960
o Screening for anticancer activity
o MTT assay
o Dual staining
o Neutral red uptake assay
o Lactic acid dehydrogenase assay
o Tryphan blue uptake assay
o DNA fragmentation
o Screening for antiviral activity
o MTT assay
o P24 Assay
o Plague assay
o Screening for antioxidant activity
o DPPH assay
o Nitric oxide scavenging activity
o Metal chelating assay
o Total antioxidant assay
o Hydrogen peroxide scavenging assay
o Screening for pigment production
o Screening for enzyme inhibitors
o Screening for mosquitocidal activity
o Screening for enzyme production
o Screening for biosurfactant production - please mention different methods
Hemolytic assay, bacterial adhesion to hydrocarbons (BATH) assay, drop collapse assay, oil
spreading assay, emulsification assay and surface tension measuremento Screening for Quorum Sensing (QSI)
o Screening for sideromycins production
o Screening for antibiofilm activity
VI. Production of bioproducts from actinobacteria
o Fermentation methods
o Agar surface fermentation
o Solid state fermentation
o Submerged fermentation
o Production using immobilized cells
o Extraction of bioactive metabolites
o Liquid liquid extraction
o Solid liquid extraction
o Precipitationo Production optimization
o Classical one-factor-at-a-time method
o Plackett-Burman model
o Response Surface Methodology (RSM)
o Kinetics of antibiotic production - add few more kinetic experiments
VII. Eliciting cryptic metabolite productiono OSMAC approach
o Inducing cryptic metabolite production using elicitors
o Epigenetic modifiers
o Microparticle enhanced cultivation
VIII. Metabolite profiling of actinobacterial extracts
o TLCo HPTLC
o UV spectral analysis
o GC-MS analysis
o LC-MS analysis
IX. Bioassay guidedisolation and characterization of metabolites from actinobacteria
o Bioassay guided isolation
o Preparative TLC
o Column chromatography
o Preparative HPLC
o Characterization of bioactive metabolites from actinobacteria
o Physico-chemical
¿ Colour, consistency, solubility
¿ Melting point analysis
¿ Chemical screening¿ Test for functional groups
¿ CHN analysis
o Spectral analysis - UV, IR and NMR
o XRD analysis
X. Evaluation of actinobacteria for environmental applications
o Heavy metal resistant/accumulation
o Pesticide degradation (qualitative & quantitative)
o Dye decolourization (qualitative & quantitative)
o Plastic degradation(qualitative & quantitative)
o Bioleaching of heavy metals from e-waste
o Anticorrosion activity
o Antibiofouling activity - antimicrobial, inhibition of mollusc foot adherence, inhibition of
algal spore germination, field testing
o Screening of Co2 sequestration
XI. Evaluation of actinobacteria for agricultural applications
o Pre-experimental screening
o Antagonistic activity against phyto pathogens
o PGP properties (IAA production, siderophore production)
o Soil fertility enhancement (N2 fixation, P & Zn solubilisation, extracellular enzyme
production)
o Experimental screening
o Non-pathogenic test in plants
o Green house experiment
o Field experiment
XII. Evaluation of actinobacteria for aquaculture applications
o Preexperimental screening
o Antagonistic activity
o Acid tolerance
o Bile tolerance
o Cell adherence property
o Non haemolytic property
o pH tolerance
o Experimental screening
o Non pathogenic test
o Lab scale challenging test for probiotic properties & response measurement
XIII.Evaluation of actinobacteria for nanoparticle synthesis
o Screening of actinobacteria for nanoparticle synthesis using
o Biomass
o Cell free supernatanto Extracts
o Characterization of nanoparticles
o Spectral
o Microscopic
XIV. Determination of cytotoxicity of actinobacterial extracts & metabolites
o Cell line based assays
o Zebra fish model
o In vivo Animal model
Notă biografică
Dr. Ramasamy Balagurunathan has been working for the past three decades exclusively in actinobacteria research for production of bioactive compounds. He has obtained his Doctoral degree in Marine Biology from Annamalai University for his research on bioactive compounds from marine Streptomyces. He pursued his post doctoral research on rare actinobacteria in reputed research Institutes in China and Malaysia. In 2019, he has obtained Doctor of Science degree in Microbiology from Periyar University for his research contribution on Bioprospecting of actinobacteria from rare ecosystems in India. He started his teaching career in 2000. He has handled eleven research projects on actinobacteria worth around Rs. 2.5 Crore. His research team discovered novel anti–TB and anti–HIV molecule from marine Streptomyces sp. R2 and named as “TRANSITMYCIN”. He is the Co-inventor in the US & Indian patent filed on Transitmycin. He has isolated Leptomycin – an anti-leptospiral compound from marine actinobacteria. With the goal of developing a centre for collection of actinobacteria from less explored habitats such as marine, desert, forest and mountain ecosystems, presently his team has isolated and are maintaining more than 500 actinobacteria isolated from various extreme environments including Thar desert, Himalayas, western Ghats, rubber forest, Andaman & Nicobar islands, which are the sources for production of antibiotics and other high value products. He has authored 145 publications including research articles, reviews, books, book chapters and magazine articles around the single focused aspect of actinobacteria for various applications like bioactive compound and bio-nanotechnology. He has an H index 19 and I10-index 28. He has guided 80 M. Sc., 25 M. Phil. Projects, 13 Ph.D thesis awarded and is guiding 6 Ph.D., scholars and 1 Post-doctoral fellow.
Dr. Manikkam Radhakrishnan is a microbiologist pursuing research on Ecology, Systematics and Bioprospecting of actinobacteria from under studied ecosystems across India for the past two decades. He obtained his Doctoral degree in Microbiology from National Institute for Research in Tuberculosis under University of Madras on “Antitubercular metabolites from Desert soil Streptomyces”. After his Post graduation, he started his career as Assistant Professor in Microbiology. He is passionate about research and joined at Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology in the year 2013. He set up a consortium of >500 actinobacterial cultures under his headship in Bioprospecting division. He has isolated a novel chromopeptide molecule, Transitmycin which is effective against TB and HIV, from a novel marine Streptomyces species R2 isolated from Coral reef ecosystem of South India.. At present he is working on Diversity and Taxonomy of Psychrophilic and psychrotolerant actinobacteria from Indian sector of Southern Ocean (Antarctic). He is participating as the scientific member (Microbiology) in the 11th Indian Scientific Expedition to Southern Ocean. He has authored 110 publications including research articles, reviews, book chapters and magazine articles and has also three patents on actinobacteria to his credit. It is his vision that this gold mine of Actinobacterial resource should be best exploited for the welfare of humans, animals, aquaculture and environmental applications since it has proven therapeutic potential and translational scope.
Dr. Thangavel Shanmugasundaram has completed his Masters in Microbiology with gold medal from Periyar University. He has received prestigious INSPIRE fellowship from DST-SERB to pursue his doctoral degree on Exploring magnesite soil actinobacteria for metal and metal alloy nanoparticles for its biomedical and environmental applications. He also received Best Ph.D Thesis award for his meritorious Doctoral thesis. He has published 31 research articles on actinobacteria in high impact. Based on his outstanding publications on actinobacteria and nanotechnology he received DST-National Post doctoral fellowship to work on actinobacteria and its enzymes for nanoparticle synthesis and applications at the DRDO Bharathiar University Centre for Life Sciences, He has a great scientific acumen and aptitude and dedicated research in the field for a decade.
Dr. Venugopal Gopikrishnan is working in the field of actinobacterial research for the past 8 years. He obtained his Doctoral degree from Actinobacterial Research Lab, Department of Microbiology, Periyar University. During his doctoral degree, he isolated about 200 actinobacterial cultures from marine and mangrove ecosystems of South India. He has identified Quercetin and Taxifolin two active metabolites from two potential Streptomyces sp PE7 and PM33 which have showed promising antifouling and antimicrobial activity. The active molecules have evidenced good antimicrobial, anti TB and anti-cancer activities. He started his career as scientist in Bioprospecting division of Centre for Drug Discovery and Development at Sathyabama Institute of Science and Technology. At present he is continuing his research on exploring the fish gut associated actinobacteria for quorum sensing inhibitors and antibiofilm compounds. He has published 29 articles including research articles, reviews, book chapters and magazine articles. His aim is to explore the host or symbiotic actinobacteria marine and terrestrial organisms for bioprospecting purposes.
Dr. Joseph Jerrine has completed her under graduation with gold medal from St. Ann’s College, Visakhapatnam, Andhra University majoring in Biochemistry, Botany and Chemistry. She has done her Masters in Life sciences from Pondicherry Central University. She also cleared GATE 2000 with 98.6 percentile and UGC-NET. She obtained the Doctor of Philosophy from University of Madras in 2011 for her research on Human Pappiloma Virus (HPV) 16 & 18 with respect to cervical cancer. She is the recipient of the ICMR –Senior Research Fellowship for “Establishment of National Database on TB”. Further, she has been awarded ICMR-Post Doctoral Fellowship for “Anti TB metabolites from marine Streptomycetes. She has published 40 research articles on actinobacteria and plants with reference to anti TB and other biomedical activities, cancer and asthma. She is recipient of three research projects from central and state funding agencies to work on actinobacteria with special reference to anti TB metabolites. Presently she is working as Scientist D and heading the Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology, TamilNadu with focused research on bioprospecting of actinobacteria and plants from rare ecosystems in India with reference to anti TB and anti cancer molecules.
Dr. Manikkam Radhakrishnan is a microbiologist pursuing research on Ecology, Systematics and Bioprospecting of actinobacteria from under studied ecosystems across India for the past two decades. He obtained his Doctoral degree in Microbiology from National Institute for Research in Tuberculosis under University of Madras on “Antitubercular metabolites from Desert soil Streptomyces”. After his Post graduation, he started his career as Assistant Professor in Microbiology. He is passionate about research and joined at Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology in the year 2013. He set up a consortium of >500 actinobacterial cultures under his headship in Bioprospecting division. He has isolated a novel chromopeptide molecule, Transitmycin which is effective against TB and HIV, from a novel marine Streptomyces species R2 isolated from Coral reef ecosystem of South India.. At present he is working on Diversity and Taxonomy of Psychrophilic and psychrotolerant actinobacteria from Indian sector of Southern Ocean (Antarctic). He is participating as the scientific member (Microbiology) in the 11th Indian Scientific Expedition to Southern Ocean. He has authored 110 publications including research articles, reviews, book chapters and magazine articles and has also three patents on actinobacteria to his credit. It is his vision that this gold mine of Actinobacterial resource should be best exploited for the welfare of humans, animals, aquaculture and environmental applications since it has proven therapeutic potential and translational scope.
Dr. Thangavel Shanmugasundaram has completed his Masters in Microbiology with gold medal from Periyar University. He has received prestigious INSPIRE fellowship from DST-SERB to pursue his doctoral degree on Exploring magnesite soil actinobacteria for metal and metal alloy nanoparticles for its biomedical and environmental applications. He also received Best Ph.D Thesis award for his meritorious Doctoral thesis. He has published 31 research articles on actinobacteria in high impact. Based on his outstanding publications on actinobacteria and nanotechnology he received DST-National Post doctoral fellowship to work on actinobacteria and its enzymes for nanoparticle synthesis and applications at the DRDO Bharathiar University Centre for Life Sciences, He has a great scientific acumen and aptitude and dedicated research in the field for a decade.
Dr. Venugopal Gopikrishnan is working in the field of actinobacterial research for the past 8 years. He obtained his Doctoral degree from Actinobacterial Research Lab, Department of Microbiology, Periyar University. During his doctoral degree, he isolated about 200 actinobacterial cultures from marine and mangrove ecosystems of South India. He has identified Quercetin and Taxifolin two active metabolites from two potential Streptomyces sp PE7 and PM33 which have showed promising antifouling and antimicrobial activity. The active molecules have evidenced good antimicrobial, anti TB and anti-cancer activities. He started his career as scientist in Bioprospecting division of Centre for Drug Discovery and Development at Sathyabama Institute of Science and Technology. At present he is continuing his research on exploring the fish gut associated actinobacteria for quorum sensing inhibitors and antibiofilm compounds. He has published 29 articles including research articles, reviews, book chapters and magazine articles. His aim is to explore the host or symbiotic actinobacteria marine and terrestrial organisms for bioprospecting purposes.
Dr. Joseph Jerrine has completed her under graduation with gold medal from St. Ann’s College, Visakhapatnam, Andhra University majoring in Biochemistry, Botany and Chemistry. She has done her Masters in Life sciences from Pondicherry Central University. She also cleared GATE 2000 with 98.6 percentile and UGC-NET. She obtained the Doctor of Philosophy from University of Madras in 2011 for her research on Human Pappiloma Virus (HPV) 16 & 18 with respect to cervical cancer. She is the recipient of the ICMR –Senior Research Fellowship for “Establishment of National Database on TB”. Further, she has been awarded ICMR-Post Doctoral Fellowship for “Anti TB metabolites from marine Streptomycetes. She has published 40 research articles on actinobacteria and plants with reference to anti TB and other biomedical activities, cancer and asthma. She is recipient of three research projects from central and state funding agencies to work on actinobacteria with special reference to anti TB metabolites. Presently she is working as Scientist D and heading the Centre for Drug Discovery and Development, Sathyabama Institute of Science and Technology, TamilNadu with focused research on bioprospecting of actinobacteria and plants from rare ecosystems in India with reference to anti TB and anti cancer molecules.
Textul de pe ultima copertă
This comprehensive laboratory manual describes the various protocols involved in Actinobacterial research. The content is divided into fifteen major sections, each of which is further divided into sub-sections describing the respective aim, principles, materials & methods, protocol, expected results and diagrams. Readers will find essential protocols for e.g. sample collection, isolation, characterization, analysis, profiling and evaluation of Actinobacteria for various applications. Gathering all relevant protocols concerning Actinobacteria, and written by a team of experienced Actinobacterial researchers, it is the first book of its kind.
Caracteristici
Deals comprehensively with all the relevant laboratory techniques in Actinobacterial research Draws attention to evaluation of Actinobacteria in diverse applications like metabolite synthesis, bioenergy synthesis, aquaculture, agriculture Provides diagrams, graphs, troubleshooting tips, to enable readers to replicate the experiments