Semester I

MB P101 BACTERIOLOGY AND VIROLOGY

Part I - Bacteriology

Part II – VIROLOGY

6. General Virology: Brief outline on discovery of viruses, origin of viruses, nomenclature and classification of viruses- Baltimore classification; distinctive properties of viruses; morphology and ultra structure; capsids and their arrangements; types of envelopes and their composition-viral genome, their types and structures; virus related agents (viroids, prions).

7. General Methods of Diagnosis and Serology. Cultivation of viruses in embryonated eggs, experimental animals, and cell cultures; primary and secondary cell structures; suspension cell cultures and monolayer cell cultures; cell stains, cell lines and transgenic systems; serological methods, assay of viruses - Physical and chemical methods (Proteins, nucleic acid, radioactivity tracers, electron microscopy) - Infectivity assay (plaque method, end point method) - Infectivity assay of plant viruses. 

8. Bacterial Viruses

Bacteriophage structural organization; life cycle; discovery, identification, nomenclature - Lysogenic and Lytic, Bacterial transformation, host cell restriction transduction one step growth curve; transcription; DNA replication; elipse phase; phage production; burst size; lysogenic cycle, application in bacterial genetics; Bacteriophage based vectors for cDNA and genetic libraties. Brief details on M13, Mu, T3, T4, and Lambda, P1.

9. Plant Viruses

Classification and nomenclature; effect of viruses on plants; common virus disease of plants; paddy, cotton, tomato, and sugarcane, life cycle; type species of plant viruses like TMV, Cauliflower mosaic virus and potato virus X; transmission of plant viruses with vectors (insects, nematodes, fungi) and without vectors (Contact, seed and pollens); diagnostic techniques in seeds; seed stocks and diseased plants (seed morphology, seedling symptomatology, indicator plants, prevention of crop loss due to virus infection - virus - free planting material; vector control.

10. Animal Viruses
Classification and nomenclature of animal human viruses; epidemiology, lifecycle, pathogen city, diagnosis, prevention and treatment of:
RNA Viruses: - Paramyxoviridae (measles), Orthomyx0viridae influenza), Rhabdoviridae (rabies), filoviridae (Ebola virus), Retroviridae (HIV), Reoviridae (rotavirus), Picorna viridae (poliovirus & FMDV of cattle), Flaviviridae (Dengue virus).

Part I - PHYCOLOGY

1. Distribution morphology and classification of Algae; Isolation from soil; algal ecology and Biotechnology-

Cyanobacteria: Distribution, thallus construction and reproduction in general, Ultrastructure of typical cyanophycean cell; Symbiosis; Economic importance of cyanobacteria; stydy of structure and reproduction of Microsystis, Gloesocapsa, spirulina, Oscillatoria, Rivularia, Nostoc, Anabaena and Scytonema. Media and methods used for culturing algae, measurement of algal growth, strain selection and large scale cultivation; applications - therapeutic uses, heavy metal removal, use of immobilized and labeled algae, economic importance of algae.

Part II - MYCOLOGY

2. Historical introduction to Mycology; Evolutionary tendencies in lower fungi- salient features and examples of Divisions Myxomycota - Acrasiomycetes, Hydromyxomycets, Myxomycetes, Plasmodiophoromycetes; Zoosporic fungi - Chytridiomycetes, Hyphochytridiomycetes, Oomycetes, Zygomycotina - Zygomycetes, Trichomycetes- Study of higher fungi - salient features and examples of Divisions Ascomycotina- Hemiascomycetes, Plectomycetes, Pyrenomycetes, Discomycetes, Laboulbeniomycetes, Loculoascomycetes; Basidiomycotina - Teliomycetes, Hymenomycetes; Deuteromycotina - Hyphomycetes, Coelomycetes, Blastomycetes.

3. Structure of Fungal cells and growth - Hyphae and non-motile unicells, motile cells, spores, dormancy, growth of population and colonies, mechanics of growth in fungi, Measurement and Kinetics of growth, nutritional and environmental requirements; effect of environment on growth - pH, temperature; prevention of fungal growth.

4. Fungi & Ecosystem - substrate groups - saprophytic, parasitic, Keratinophilic, coprophilous; substrate successions, parasitism, mutualism and symbiosis with plants and animals.

5. Heterothallism, Sex hormones in fungi, Physiological specialization, Phylogeny of fungi; fungal diseases - mycoses, systemic and subcutaneous, candidiasis, pnemocystis, blastomycoses, dermatomycoses.

MB P 103 MICROBIAL DIVERSITY AND CELLULAR MICROBIOLOGY

Part I - MICROBIAL DIVERSITY

1. Introduction to microbial diversity - distribution - abundance - ecological niche. Oxygenic photosynthetic microbes - anoxygenic photosynthetic microbes- oxidative transformation of metals - sulphur oxidation, iron oxidation, ammonia oxidation and hydrogen oxidation. Adaptation to the nutritional and physical environment.

2. Bacterial Systematics and nomenclature: Biological Classification and the Linnaean system, Nature System of Classification. Taxon, strain, Binomial nomenclature, Bacterial species. Haeckal's Three-kingdom classification, Whittakers Five kingdom classification. Molecular phylogeny, Three-domain concept of Carl Whose. Criteria used for classification. Classification according to Bergeys Manual of Systematic Bacteriology. Cyanobacteria, Phochlorons and Cyanelles. Recent trends in Microbial Taxonomy-

a) Chemo taxonomy [cell wall components, lipid composition, isoprenoid - quinines, cytochrome composition, DNA homology, DNA-RNA homology, G+C ratio, rRNA sequencing etc., 
b) Numerical Taxonomy

c) Genetic methods in taxonomy

d) Serological methods. Bacterial nomenclature. Phylogenetic Trees - evolutionary models, homology, gene trees versus species trees, methods from Tree building, maximum likelihood, organizing data on a tree, evaluating phylogenies. Dichotomous key. International Code of Nomenclature of prokaryote.

3. Unculturable and culturable bacteria - conventional and molecular methods of studying microbial diversity. Culture dependent methods of community analysis - Dilution plating and culturing methods, community level physiological profiles. Culture independent methods of community analysis - phospholipid fatty acid analysis, nucleic acid techniques, phylogenetic analysis, fluorescent in situ hydrization (FISH)

6. Subterranean microbes - ground water contamination and microbial transformations. Biomagnification, bioaccumulation and bioremediation. Catabolic pathway of recalcitrant molecule degradation and mineralization.   

1. Introduction: Bacterial diseases. Emergence of cellular microbiology, cellular biology underlying prokaryotic and eucaryotic interactions: ultra structure.

2. Infection and cell - cell interactions; bacterial adherence; basic principles, effect of adhesion on bacteria, effect of adhesion on host cells, bacterial invasion of host cells, mechanism, consequence of invasion survival after invasion. Protein toxins: agents of diseases.

3. Prokaryotic and eukaryotic signaling mechanisms: eukaryotic cell-to-cell signaling endocrine signaling, cylikins prokaryotic signaling: quorum sensing and bacterial pheromones intracellular signaling. Concepts of receptor, example of hormone, growth factors, Histidine kinase signaling in bacteria, Tyrosine kinase coupled receptor in Proteins and G-proteins receptor coupled signaling, Mitogen activated protein kinase pathway, Signaling pathways.  

MB P 104  BIOPHYSICS AND BIOSTATISTICS

SECTION A: BIOPHYSICS

1. Chemical and physical forces involved in chemical bond formation between atoms and molecules; mechanisms of bond formation based on electronic orbitals; sigma, pibonds, covalent, ionic, electrostatic, co-ordinate bonds; hydrophobic and Vanderwaals interaction and their properties, chemical bonds between carbon and carbon, Carbon and Nitrogen and Carbon with other atoms such as Hydrogen, Oxygen, Phosphorous and Sulphur, Conformation of proteins and polypeptides; Reverse turns and Ramachandran plot.

2. Physical methods of determining the sizes and shapes of molecules; Hydrodynamic Properties, surface tension, diffusion, osmotic pressure, sedimentation.

3. Isotopes and Radioactivity; dosimetry; Radioactivity, delay laws, production of radio-isotopes, isotopic tracer method, assay using radioactive active substrates, biological life time, Metabolic and physiological tracer studies, dose response relationship, non-radioactive labels, labeling and detection methods using fluorescent molecules like diagnosing biotin, rhodomin and other chromogenic compounds. Cerenkov radiation; Liquid Scintillation spectrometry.

4. Instrumental methods analysis: Ultra-centrifugation, Viscometry, Flame Photometry; Hydrodynamic methods; Atomic absorption and plasma emission spectrophotometry, Chromatography, Electrophoresis, X-ray crystallography, NMR and ESR spectroscopy.

SECTION B: BIOSTATISTICS

1. Scope of statistical methods in scientific studies; samples and populations; Frequency distribution-preparation of frequency table: relative and cumulative frequencies: diagrammatic representation of frequency distribution; histograms, frequency polygons, frequency curve and ogives.

2. Measures of central tendency: Arithmetic mean, mode, median and percentiles. Measures of variability: Range, mean deviation, and analysis of variance. Standard deviation and Co-efficient of variation. Skewness and Kurtosis

3. Probability: Definitions; events; sample space; addition and multiplication rules of probability; conditional probability (simple problems).

4. Probability Distributions: Discrete, Continuous, binomial, Poisson and normal distribution; Simple correlation of regression.

5. Population and Sample: Random sample, use of table of random numbers, parameter and statistics, sampling distribution of sample means, Standard error; confidence intervals.

6. Hypothesis testing: Basic concepts and definitions, tests based on normal, student T, chi square and F distribution (no proofs)

7. Design of experiments: Basic principles of scientific experimentation: randomization, replication and local control, completely randomized and randomized block designs

Practicals

MB P 105 BACTERIOLOGY AND VIROLOGY

1. Staining Techniques: Differential, acid - fast, endospore, capsule, cell wall, cytoplasmic inclusion,
Vital Stains: Negative Staining - flagella, spore, nuclear

2. Study of Cultural Characteristics of Bacterial [Autotrophic & Heterotrophic], selective - differential media.
Growth on NA, blood agar, MRS, Chocolate agar, DCA, Mac Conkey's, Dorsett's egg growth medium.

3. Study of nutritional needs of bacterial growth at 37oC and 20o C. [Growth in presence of different sugars - C source, N source]

4. Pure culture techniques - preparation of slants, sub culturing type of streaking.

5. Biochemical tests for identification of bacteria.
Catalise, Oxidase, IMViC, motility at 37oC and 20o C, pigmentation, spreading growth, resistance to 60 C for 20m, gelatin test, urease, levan formed form glucose, H2S in TISA, H2S in lead acetate paper, coagulase, optochin sensitivity, lecithinase, nitrate reduction, acid and gas from glucose, arabinose, inositol, lactose, ONPG acid, maltose, mannitol, rhamnose, salicin, trehalose, sucrose, xylose, fructose,hippurate hydrolysis, chitin, starch, casein, Tween 80 hydrolysis, pectin, organine, dehydrolysis, lysine decarbosylase, ornithine, esculin hydrolysis.

6. Microbial growth measurements - cell count, turbidometry measurements, serial dilutions, Plate count.

7. Identification of bacteria by API system

8. Isolation of bacteriophages from sewage.

9. Isolation of bacteriophages from files.

10. One step growth curve of bacteriophages.     

MB P 106  PHYCOLOGY, MICOLOGY AND MICROBIAL DIVERSITY

1. Isolation of fungi from different substrates (Saprophytic, Parasitic, Copraphilous, Keratinophilous)

2. Study of environmental requirements of Fungi (pH, Temperature) by linear growth and biomass.

3. Study of antifungals (antibiotics/ Chemicals/ Plant extracts)

4. Study of antagonism

5. Isolation of algal pigments

6. Separation of algal pigments

7. Isolation of Cyanobacteria

8. Gel electrophoresis of Cyanobacterial pigments.

9. Isolation and quantification of Cyanobacterial pigments.

10. Enrichment and isolation of sulphate reducing bacteria.

11. Isolation of alkalophilic bacteria from different soil samples.

12. Isolation of Thermophilic Anaerobic cellulolytic bacteria.

13. Effect of temperatures on growth of different fungal cultures.

14. Estimation of Alkaline protease production by Bacillus licheniformis in solid state fermentation.

15. Demonstration of Biofilm formation on different substrates by pseudomonas aeruginosa and klebsiella pneumoniae.

16. Isolation and characterization of acidophiles and halophiles.

17. Isolation of bacteria from Groundwater samples.           

Semester II

1. Microbial Nutrition - Organic and Inorganic nutrients: Nutritional types Basic aspects of energetics - entropy - enthalpy, Electron transport chain, artificial electron donors un couplers - energy bond - phosphorylation

2. Brief account of photosynthetic and accessory pigments-Chlorophy-II-bacterial Chlorophy - II - rhodospin - carotenoids - phycobiliproteins; carbohydrates-anabolism - autotrophy - oxygenic-anoxygenic photosynthesis; autotrophic generation of ATP; fixation of CO2 - Calvin Cycle - C3 and C4 pathways; chemolithotropy - sulphur - iron - hydrogen - nitrogen oxidations, methanogenesis - luminescence; Biosynthesis of fatty acids; pigments and secondary metabolites. Co-ordinated control of metabolism;

3. Enzymes - definition, structure, Enzymes as biocatalysts, properties and classification, specificity, active sites, Coenzymes: Activators and inhibitors, activity unit, Isozymes, Enzyme Kinetics (negative and positive cooperatively); Michaelis Menten equation for simple enzymes inhibition, allosterism, kinetic analysis of allosteric enzymes, principles of allopsteric regulation, Ribozyme and abzyme.
Respiratory Metabolism: Embden Meyerhoff Parnas pathway: Entner Doudoroff pathway; glyoxalate pathway; krebs cycle; Oxidative and substrate level phosphorylation, reverse TCA cycle; gluconeogenesis, Pasteur effect; fermentation of carbohydrates-homo and heterolactic fermentation.

4. Assimilation of nitrogen-dinitrogen, nitrate nitrogen, ammonia, synthesis of major amino acids, polyamines; synthesis of polysaccharides-peptidoglycan-biopolymers as cell components; endospores-structure, properties and germination. Biosynthesis of purines and pyrimidines.

5. Sporulation and morphogenesis-hyphal and yeast forms and their significance Multicellular organization of selected microbes; Dormancy   

MB P 202  MICROBIAL GENETICS   

1. Structural polymorphism of DNA: DNA structure: A, B and Z DNA, super coiled DNA, DNA binding proteins, DNA replication in Prokaryotes, the rolling circle replication, semi conservative replication, primer or template, origin of replications, relication fork-leading and lagging strands, enzymes involved at different steps of replicationh, DNA proof reading and repair mechanisms. Fine structure of gene, DNA amplification and rearrangements. The law of DNA constancy and C-Value paradox.

2. Gene as unit of mutation, Molecular basis of spontaneous and induced mutations and their role in evolution; Mutagens, types of mutations, Transposon mutagenesis, site directed mutagenesis, AME's test; environmental mutagenesis and toxicity testing.

3. Genetic recombination at molecular level: Genetic recombination in bacteriophages and E.coli, Synapsis of homologous duplexes, breakages and re-union, role of Rec A in recombination, generalized & specialized transduction, transformation and conjugation, Legitimate & illegitimate recombination gene conversion, overview of bacterial genetic map.

4. Gene transfer mechanisms-Bacterial transformation; Host cell restriction; Transduction; Complementation; Conjugation and transfection, Mechanisms and applications, Genetic analysis of microbes, Bacteria and Yeast.

5. Plasmids, F-factors - description and their uses in genetic analysis, Colicins and Col Factors R plasmids as vectors for gene cloning, Replication of selected plasmids: compatibility, Transposons and their use in genetic analysis.

6. Bacteriophages: Lysogeny and lytic cycle in bacteriophages, Life cycle and their uses in microbial genetics. Lytic phages-T7 and T4, Lysogenic phages I and P1. M13 and fX174.   

MB P 203  IMMUNOLOGY

1. Immune System and Immunity

History of immunology; Structures, composition and functions of cells and organs involved in immune system - T- cells, B-cells, macrophages, antigen-processing cells, Eosinophls, Neutrophils, Mast cells and killer T - cells; microbial infections and immune reponses - innate Immunity, acquired Immunity; Clonal nature of immune response; Immunohaematology- blood groups, blood transfusion and Rh incompatibilities.

2. Antigens and Antibodies

Antigens - structure and properties - types - Iso and all antigens - haptens; adjuvants - antigen specificity. Immunoglobulins - Structure - heterogenecity - types and subtypes - properties (physico-chemical and biological); Complement system - Structure, components, properties and functions of complement; Complement pathways and biological consequences of complement activation. Generation of immunological diversity; Effector mechanisms.

3. Antigen-antibody Reactions

In vitro Methods-Agglutination, precipitation, complement fixation, immunofluorescence, immuncelectrophoresis, ELISA, RadioImmunoassays; In vivo Methods: skin tests and immune complex tissue demonstrations. Applications of these methods in diagnosis of microbial infections.

4. Major Histocompatibility Complex and Tumer Immunology

Structure and functions of MHC and HL-A systems. Gene regulation and Ir-genes HL-A and tissue transplantation - Tissue typing methods for organ and tissue transplantations in humans; graft versus host reaction and rejection; autoimmunity Tumor immunity - tumor specific antigens, Immune response to tumors, immunodiagnosis of tumors - detection of tumor markers - alphafoetal proteins, carcinoembryonic antigen etc., Genetic Control of Immune response.

5. Hypersensitivity Reactions

Definition of allergy, Anibody-mediated Type L anaphylaxis; Type II. Antibody dependent cell cytotoxicity; Type III. Immune complex mediated reactions; Type IV. Cell mediated hypersensitivity reactions. The respective symptoms, immunological methods of their diagnosis. Lymphokines and cytokines - their assay methods. Immunological tolerance.

6. Immunization

Vaccines (Conventional; submit vaccines; DNA vaccines) toxoids; antisera; polyclonal and monoclonal antibodies; hybridoma technology to produce monoclonal antibodies; Catalytic monoclonal antibodies; common immunizations: small pox, DDT, polio, measles, hepatitis-B, vaccines from plants-banana, watermelon etc (plantibodies)   

MB P 204  COMPUTER APPLICATIONS & BIOINFORMATICS   

1. Computer basics, Operating systems, Windows and Unix

2. Hardware, Software, disk operating system, multimedia network concepts

3. C-programming: Object Oriented Programming, tabulation of date, & graphical representation, flow charts

4. Internet, Local Area Network, Wide Area Network, Web Servers, Web Access, Security HTML, HTTP & XML concepts.

5. Overview of Bioinformatics - Database types, sequence databases - nucleotide and protein sequence databases, Primary and secondary databases, Gene Bank, Structure databases- Protein Data Bank (PBD), visualization of structural information

6. Genomics and the genome project, computer tools for sequence analysis, finding and retrieving sequences, similarity searching, Pairwise and multiple alignment

7. Structure function relationships

8. Sequencing and sequence assembling using computers phylogenesis

PRACTICALS

MB P 205 MICROBIAL PHYSIOLOGY

1. Determination of Growth - Growth curve and generation time.

2. Colorimetric of Spectrophotometric estimations of proteins DNA, RNA and sugars.

3. Qualitative analysis of Cyanobacterial photosynthesis.

4. Estimation of microbial enzymes-amylase, protease, invertase, cellulase, lipase, catalase and phosphatase.

5. Lipids, Saponification value of fat, iodine number of fatty acids.

6. Determination of Km and Vmax.

7. Polyacrylamide Gel Electrophorsis (SDS - PAGE) for proteins.

8. Extraction and seperation of aflatoxins by paper chromatography

MB P 206  IMMUNOLOGY & MICROBIAL GENETICS   

1. Slide agglutination tests

2. Precipitation test

3. Blood Grouping and RH typing.

4. Serological tests of diagnostic importance 

5. Mutagenesis: Identification and Isolation of fungal and bacterial mutants - Chemical and UV

6. Study of Replica plating technique.

7. Agarose gel electrophoresis

8. Isolation of plasmid DNA, genomic DNA

9. Phage titration

10. Ame's test for detecting possible chemical carcinogens.

11. ELISA

12. Demonstration of bacterial transformation.

13. Demonstration of nonspecific resistance to bacteria.

14. Determination of bactericidal activity of normal serum.   

BT P301-Molecular Biology

1. DNA Replication: Initiation of replication of ssDNA; retrovirus ant their unique mode of DNA synthesis, relationship between replication and cell cycle, inhibitors of DNA replication (blocking precursor synthesis, nucleotide polymerization, altering DNA structure) DNA damage and repair : Types of DNA damage – deamination, oxidative damage, alkylation, pyrimidine dimmers; replicative pathways- photo-reactivation, excision repair, post replication repair, SOS repair, methyl directed mismatched repair, very short patch repair.

2.Structural features of rRNA, tRNA  and mRNA  and their relation to function. Initiator and elongator class of tRNA, ribosome binding site on the mRNA and corresponding site on rRNA. Peptidyltransferase activity of 23srRNA. 

3. Transcription: RNA polymerase in prokaryotes- its molecular composition, role of each component of RNA polymerase, mechanism of transcription- initiation, elongation and termination, global regulatory responses, heat shock responses, stringent response and regulation by small molecules such as ppGpp and cAMP, regulation of rRNA and tRNA synthesis, inhibitors of RNA synthesis and their mechanism of action, polycistronic and monocistronic RNAs; Control of transcription by interaction between RNA polymerase and promotor regions, use of alternate sigma factors; controlled termination – attenuation and anti-termination.

4. Translation: Role of ribosomes and different types of RNA in protein synthesis; basic feature of genetic code; Amino acid activation, mechanism of intitiation, elongation and termination. Inhibitors of protein synthesis; synthesis of transport proteins on membrane bound ribosome; signal hypothesis; invitro transcription and system translation.  

5. Regulation of gene expression in prokaryotes: Transcriptional control; enzyme induction and repression, constitutive synthesis of enzymes. The operon concept, catabolic repression, instability of bacterial RNA, inducers and corepressors; Negative gene regulation- E.Coli lac operon; regulation by attenuation- his and trp operons, anti termination- N protein and rr1 sites, DNA binding protein, enhancer sequences, identification of protein binding site on DNA Maturation and processing of RNA- methylation, cutting and trimming of rRNA, capping, polyadenylation and splicing of Mrna; cutting and modification of tRNA degradation system, catalytic RNA, GI and GII in trans splicing, Rnase P .

6. Genetic control and lambda phage. Switching between lytic and lysogenic cycles. Control of transcription. RNA polymerase, repressor, Cro gene; control of integration and excision.

MB P 302 Recombinant DNA Technology

1.  Recombinant DNA Technology – introduction, definition, scope and importance

2.Restirction endonucleases – type, nomenclature, recognition sequences, cleavage pattern

DNA ligases – properties and function of T4 DNA ligase and NAD-dependant DNA ligase of E, coli, ligation techniques, 

Vector-general characteristics, cloning and expression. Types of vectors- plasmids (pBR 322, PUC Vectors, Ti), Phages, Cosmids, Phagemids, shuttle vectors, ARS, Mini-Chromosome, yeast artificial chromosomal vectors (YAC). Promotor probe vectors, vectors for library construction.

3.Cloning strategies – cloning and selection of  individual gene libraries: cDNA libraries and genomic libraries, cosmid, bacterial artificial library, shot gun library, agrobacterium mediated gene transfer.  

5. Ligation and transformation : DNA ligase, properties and function of T4 DNA ligase and NAD dependent DNA ligase of E coli; ligation techniques, introduction of vectors into suitable hosts, selection of recombinant clones, purification of recombinant DNA.

6. PCR methods and application, DNA microarray- Printing as oligonucleotides and PCR product on glass slides, nitrocellulose membranes. Whole genome analysis for global patterns of gene expression, fluorescent labeled cDNA and labeled RNA probes. Analysis of single nucleotide polymorphisms using DNA chips. Advantages and disadvantages of DNA  microarray.

7. DNA sequencing methods- Dideoxy and chemical methods; sequence assembly, automated sequencing, genome sequencing and physical mapping of genomes.

8. Chemical synthesis of genes: Phosphodiester, phosphotriester, phosphite triester approaches. Enzyme synthesis of DNA; Application of synthetic oligonucleotides, synthesis of complete gene.

9.Application of rDNA technology in  development of various resistance and tolerance. Production of growth hormones, interferon, insulin, recombinant vaccines, gene therapy, antisense RNA technology. Requirement of  recombinant molecules in health, pharmaceuticals, agriculture and industrial sectors, research labs.

MB P303  Environmental Microbiology

1. Aerobiology: droplet nuclei, aerosal, assessment of air quality- solid and liquid impingement method. Brief account of air born transmission of microbes (viruses, bacteria, fungi and actinomycetes), microbiology of indoors and outdoor environments, diseases caused by air born microbes (infection, allergy and non allergic) and preventive measures. Airborne endotoxin in occupational environments, their detection and enumeration. Biohazards caused by endotoxins.

2.Aquatic microbiology: water ecosystem- fresh water (ponds, lakes, streams), marine habitats (estuaries, mangroves, deep  sea, hydrothermal vents, salt pans, coral reefs). Zonation of water ecosystems; upwelling, eutrophication; food chain in aquatic ecosystems, portability of water, microbial assessment of water, water purification. Brief account of major water borne diseases and their control measures.

3. Soil microbiology: characterization  based on physical and chemical characteristics, microorganisms in various soil types: rhizosphere, rhizoplane and phyllosphere. Microbial interactions- symbiosis, mutualism, commensalisms, competition, amensalism, synergism, parasitism,  predation; biogeochemical cycle- (carbon, nitrogen, phosphorus and sulphor) and  microorganisms.

4. Waste  treatment : solids and liquid wastes and their characterization, Treatment- physical, chemical, biological, solid waste  treatment: Saccharification, gasification, composting; Utilization of solid wastes- food  ( SCP, mushroom, yeast) : fuel ( ethanol, methane), fertilizer (Composting); Effluent treatment- physical, chemical and biological, sewage treatment, activated sludge, trickling filters, oxidation ponds and ditch. Subterranean  microbes..

5. Positive and  negative  roles of microbes in environment: biodegradation of recalcitrant compounds- lignin, pesticides, hydrocarbons and plastic; bioaccumulation of metals and detoxification. Bioremediation (in situ and exsitu) of contaminated soil and water (oil slicks, pesticides, heavy metals). Biodeterioration of paper, leather, wood, textiles, mode of deterioration, organisms involved , mode of prevention, metal corrosion.    

MB P304 FOOD MICROBIOLOGY                                                                                                    60 hrs

Unit 1 Introduction: Development of food microbiology as a science and the importance of studying food microbiology.                                                                                                                                                             2 hrs

Unit 2:Food as the substrate for micro organisms: Important micro organism food (molds,yeasts, bacteria,)and their source (air,soil,water,plants and animals)                                  
                                                                                                                                                  9 hrs

Unit 3:Contamination and  spoilage: Principles of food spoilage ;Spoilage of cereals, sugar products, vegetables, fruits, meat and meat products, milk and milk products, fish and sea foods,poultry,Spoilage of canned foods; Detection of spoilage and characterization.

                                                                                                                                                   10 hrs

Unit 4:Food Peservation:Principle and methods of food preservation-low temperature, high temperature,canning,drying,use of chemicals, irradiation and food additives; Processing for heat treatment-D,Z and F values and working out treatment parameters.

                                                                                                                                                   10 hrs

Unit 5:Food borne infections and intoxication:

Bacterial-Bruella,Bacillus,Clostridium,Escherichia,Salmonella,Shigella,Staphylococcus,Vibrio,Yersinia and Listeria.

Nematodes,Protozoa,Algae,Viruses,Molds.Mycotoxin-aflatoxin,ochratoxin,trichothecenes,zealenone,ergot,alkaloids;Food borne outbreaks and lab testing procedures, Preventive Measures.                                                                                                                                                    12 hrs

Unit 6:Fermented and microbial foods: Microbial activity in foods-vegetables (olive, cucumber), meat (sausages),bread,idli,cocoa and coffee; Dairy foods-kefir, kumis, acidophilus milk: therapeutic and nutritional value of fermented foods, spoilage and defects of fermented dairy products, oriental fermented foods, their quality standard and control. Microbial foods- single cell proteins (bacterial, fungal, algal and actinomycetous), single cell oil (fungal and algal): experimental and industrial production method.                                                                                                                                                     12 hrs

Unit 7:Food and sanitation: Sanitation in manufacture and retail trade; food control agencies and their regulation, hazard analysis and critical control points (HACCP): plant sanitation- employee's health standard, waste treatment, disposal quality control                                          5 hrs

MB P305 MOLECULAR BIOLOGY & RECOMBINANT DNA TECHNOLOGY

8X 15=120hrs

1. P1 Transduction in E.coli
2. Conjugation in E.coli
3. Chromosomal mapping by interruption mating
4. Isolation of antibiotic resistant mutants by gradient technique
5. Mutagenesis with ethyl methyl sulfonate
6. Ampicillin selection for enrichment of auxotrophs
7. Screening for nonsense mutations
8. Transposon Tn 10 d Cm mutagenesis & identification of Lac mutants
9. In vivo mutagenesis of plasmid with MNNG
10. Generation of DNA fragments by restriction digestion & ligation
11. Transformation by electroporation/ CaCl₂ method
12. Western & Southern blotting
13. Induction & assay of β-galactosidase

MB P306 ENVIRONMENTAL & FOOD MICROBIOLOGY

8X 15=120hrs

1. Quantification of microorganisms in air(out door & indoor occupational environment) solid impaction & liquid impingement techniques
2. Sampling & quantification of airborne endotoxins by Limulus Amoebocyte Assay
3. Isolation, cultivation & characterization of iron & manganese reducing bacteria
4. Isolation methanogenes from enrichments
5. Isolation of Nucleic acids from environmental samples
6. Physical, chemical & microbial assessment of water & portability test for water Microbiological- Heterotrophic plate count, MPN index, presumptive, Confirmatory & completed tests, membrane filter technique for total coliforms, fecal coliforms, Clostridia, Pseudomonas
7. Study of microbial tolerance/resistance to heavy metals by agar dilution method, agar diffusion method
8. Chemical characterization of bacterial exopolymers produced in a biofilm
9. Screening of microorganisms for biodegradation of recalcitrant compounds
10. Study of microflora of industrial wastes & effluents
11. Study of milk milk product(hydrated/dehydrated) cell count by Breed's method, isolation of mastitis organisms, Yeasts & molds, determination of hygienic  quality of milk. Isolation of Salmonella, Staphlococcus
12. Study of fermented food including vegetables ( Isolation of aerobic mesophilic counts, Total coliforms Staphylococcus, Listeria, mold count)
13. Study of fruit juice, squashes beverages (viable count of yeasts molds, osmophilic count, spore count, ascospore count)
14. Examination of flour( anaerobic counts, mesophilic, thermophilic & viable counts)
15. Study of bottled water, mineral water & carbonated water( total coliform, fecal coliform, aerobic, mesophilic & psychrophilic counts, examination for Pseudomonas aeroginosa, Aeromonas.

MB P401 AGRICULTURAL MICROBIOLOGY       60 hrs

Unit 1: Decomposition of organic matter by microbes- cellulose, hemicelluloses, lignin, starch and pectin.                                                                                                                                                                            7hrs

Unit 2: Biological nitrogen fixation: mineralization and immobilization of nitrogen, nitrification, denitrification, symbiotic nitrogen fixation (rhizobium, frankia), nonsymbiotic nitrogen fixation (azotobacter, azospirillum)- VAM: nitrogenase enzyme, nif gene.                                                                                                                                              8 hrs

Unit 3: Microbiology of rhizosphere microbes in the rhizosphere and rhizoplane, methods of umeration, rhizosphere effect, factors influencing rhizosphere microbes
                                                                                                                                                     15 hrs  

Unit 4: Biofertilizers-cultivation and mass production of azotobacter, rhizobium, rhizospirillum, cyanobacteria, phosphate solubilising bacteria; carrier based inoculants, methods of application and agronomic importance. Microbiology and biochemistry of biogas production. Biopesticides-types and application. Mushroom cultivation                               
                                                                                                                                                      15 hrs

Unit 5: Plant disease: mode of entry of pathogens, symptomatology; physiology of parasitism, host pathogen relationship; control measures- physical chemical and biological: detail study of fungal (Tikka disease of groundnut, redroot of sugarcane, wilt disease), viral (tobacco mosaic virus, tomato spotted wilt, bunchy top of banana) and mycoplasma (sandal spike, grassy shoot) diseases and viroids
(potato spindle fibre disease, citrus viroidal disease). Phytotoxins and phytoelexin
                                                                                                                                                       15 hrs

UNIT-1
Classification of medically important microorganisms.normal micro flora of human &its significance,tissue tropism.

UNIT-2
Establishment,spreading,tissue damage and anti phagocytic factors;mechanism of adhesion,colonization & invasion of mucous membrane of respiratory,enteric and tracts.role of agreesins,invasinr,de polymerizing enzymes,orgonotropism,virulent variations,evasion of host defences,non-spesific host defences,toxigenisisbacterial toxins & endotoxin.

UNIT-3
Study of pathogenic bacteria- staphylococcus, streptococcus, pneumococcus, Neisseria corynebacterium, bacillus, clostridium, non –sporing anaerobes, enterobacteriaceae, mycobacterium, spirochetes, actinomycetes, rickettsiae, chlamydiae, mycoplasmaProtozoal infections-malaria and filariasis.

Unit-4
Description and classification of pathogenic fungi – their diagnosis and control, mycosis superficial and systemic mycoses, mycetoma, chromomycosis, porotrichosis, rhino spirocosis, cryptococcosis, blastomycosis, coccodiodomycosis,  histoplasmosis, oppourtunistic systemic mycoses. Mycotoxins allergic fungal diseases.

UNIT-5
General properties of viruses: virus-host interactions; pox virus, herpes virus, adeno virus, picoma virus, orthomyxo viruses, paramyxo viruses, arbo viruses, rhabdo viruses, hepatitis viruses, onco genic viruses, HIV, yellow fever, dengue, encephalitis, prion infection – CJD, kuru, viroid infection a brief note

UNIT-6
Antimicrobial therapy: various methods of testing drug susceptibility, antibiotic assay in body fluids, Brief account of available vaccines and schedules; passive prophylactic measurement, nosocomial infections-diagnosis and control.

UNIT-7
Microbial Diagnostics- Tests to identify microbial infections.

  MB P403 Microbial Technology

1. Definition and scope of microbial technology; Historical developments; economics of microbial technology; Biotechnological innovation in the chemical industry. Biocatalyst in organo chemical regulation, P/O quotients, metabolite over production and growth efficiency.

2. Industrial strains: Criteria for selection and strategies for improvement; maintanence and containment of recombinant organisms, product recovery.

3. Microbial products of commercial use: Industrial production of organic acids (Citric acid, acetic acid, gluconic acid), production of amino acids, solvents (ethanol, glycerol), antibiotics, penicillin, streptomycin), microbial polysaccharides and polyesters. 6-hydroxyalcanote production and diversification of antibodies: steroids.

4. Microbial Enzymes: Industrial production of microbial enzymes (pectinases, proteases, cellulases), use of enzymes in starch processing, cheese making, textile making, textile industries, detergent industries, leather industries, breweries, pharmaceuticals.

5. Replication of microbial systems, rationale for the design of vectors for the expression of recombinant proteins, selection of suitable promoter sequences, ribosomes binding sites, transcription terminator, fusion protein tags, purification tags, protease clevage sites and plasmid copy number, inducible expression system, determination of purity of over expressed proteins

6. Immobilized cells and enzymes: potential benefits of immobilization. Two dimensional seperation of total cellular proteins. Protein microarray.

7. Microbes in mineral recovery; metal leeching by bacteria from ores (Copper, iron, uranium).

 MB P404 FERMENTATION TECHNOLOGY                                                                                                             60 hrs

Unit 1: Concept and scope of fermentation technology                                                                                                    2 hrs

Unit 2: Fermentation in batch and continuous: Batch growth, Microbial growth kinetics, kinetics of thermal death of micro organisms, Effects of environment-temperature, pH  and high nutrient concentration, monitoring microbial growth in culture cell number, direct and indirect methods; Continuous culture system, growth kinetics, productivity and product formation. Power requirements, oxygen transfer kinetics, concepts of Newtonian and Non-Newtonian fluids. Plastic fluids, apparent viscosities, foam and antifoam.                                                                                                           10 hrs

Unit 3:Fermenters: Basicfunctions, design and components-body construction and temperature control, aeration, agitation systems, baffles. Different types of fomenters. Sterilization of fomenter,Medium and air supply. Metabolic pathways and metabolic control mechanisms, primary and secondary metabolites, aseptic inoculation and sampling methods; Scale up of fermentation process-parameters used in scale up, problems associated.           18 hrs

Unit 4:Formation media: Media formulation strategies, sources of carbon, nitrogen, vitamins and minerals; role of buffers, precursors, inhibitor and inducers.
12 hrs

Unit 5:Downstream processing: Objectives and criteria, foam sepration, precipitation methods, filtration, centrifugation, cell disruption methods, liquid-liquid extraction, membrane filtration, chromatography, drying devices, crystallization .solvent recovery
15 hrs

Unit 6:Quality control of fermented products-chemical and microbial analysis
3 hrs

 PRACTICALS

MB P405 AGRICULTURAL AND MEDICAL MICROBIOLOGY

                                                                                                            8*15=120 hrs   

1. Isolation of cellulose,hemicellulose,starch,lignin,pectin degrading microorganism.
2. Isolation of Symbiotic and Non-symbiotic nitrogen fixing micro organisms.
3. Demonstration of nitrogen fixation by nitrate reduction.
4. Assay of Biofertilizers included in the theory.
5. Isolation of methanogens and hydrogen producing bacteria.
6. demonstration of biogas production using different substrate like cattel ung,water hyacinth,sewage
7. Demonstration of effect of biopesticide(Bacillus thuringinsis)on lepidopteran larvae.
8. Mushroom cultivation and evaluation of protein content.
9. study of host parasite relationship of plant pathogens included in the syllabus
10. different staining technique
    1)      ziehl- Neelsen method for AFB
    2)      Flurochorome staining
    3)      Leishman's staining
    4)      Geimsa's staining
    5)      Special staining methods to demonstrate granules, capsules and spores.
11.   Anaerobic culture method for Anaerobes of clinical importance.
12. Presumptive identification of pathogen using colony morphology on selective/differential/selective- differential /enrichment media
13. Isolation and characterization of clinically significant species of staphylococcus,streptococcus,corneybacterum,Bacillus,Nocardia,Neisseria,Enterobacteriaceae,Vibrio,Pseudomonas,Aeromonas.
14. Drug susceptibility testing by various methods according to NCCLS.Determination of MIC  for selected antibiotics(Kirby-Bauer methods,T test,checkerboard method)
15. Demonstration of checkerboard synergy tests for combination of antibiotics.
16. Diagnostic immunologic principles and method      

- Precipitation Methods                        -Immunociffusion
                                                            -Immunoelectrophoresis                                                        

-Agglutination methods                         -Widal test,VDRL test
                                                           -Haemagglutination
                                                            -ELISA method

 MB P406 MICROBIAL AND FERMENTATION TECHNOLOGY

                                                                                                            8*15=120hrs

1. demonstration of hydrogen evolution by bacteria using biomass as substrate
2. study of the microbial aspects of anaerobic digestion
3. entrapment of bioicatalysts including microbial cells using gel method
4. demonstration of metal leaching, lab  extraction
5. Production of citric acid from A.niger,T.viridae,P.luteum.
6. production of peniilin and ethanol by small scale solid state fermentations
7. Detection and quantification of pigments produced by certain fungi and bacteria (Penicillium,Chrysogemum,Serratic marcescens,Pseudomonas spp.
8. Detection and quantification of siderophore produced by pseudomonas spp.
9. Microbial assay of antibiotics, vitamins, aminoacids.
10. sterility testing of fermented, pharmaceutical products
11. Alcoholic fermentation and calculation of total and volatile acidity.
12. role of yeast in bread making
13. Bradford Protein asay and calculation of protein concentrations
14. immunological analysis of the purified protein
15. Production of ά-amylase, using Aspergillus oryzae/Bacillus licheniformis using wheat bran as substrate in Small Scale Solid State Fermentation and its asay.
16. production of protein by Aspergillus niger using wheat bran,dried skim milk by SSF and its assay
17. Industrial/Institutional visits-report to be submitted