Microsoft word - iii microbiology.doc

Solapur University, 
B.Sc.­III Microbiology 
Syllabus 2009‐ 2010 
(Paper -V)
Microbial Taxonomy and Genetics
Section I: Microbial Taxonomy and Bioinformatics
Bacterial nomenclature and classification
A) Comparative study of Bacteria, Archaea and Eucarya. . B) Classification of prokaryotic organisms – an overview, C) Principles of bacterial nomenclature. D) Microbial taxonomy Classical and Genetic approaches: i) Classical approach:-Morphological, Physiological, metabolic, Ecological and Serological. ii) Genetic approach :-( G+C) content and rRNA sequencing in identification of microorganisms. Classification of viruses
A) Viral classification, LHT system and as per international committee B) Isolation, cultivation, purification and enumeration of viruses Reproduction of bacteriophages
B) Reproduction of ds DNA/ ss DNA / RNA bacteriophages C) Temperate phages and lysogeny of λ phages Types of extremophiles, General characteristics of and their role in
respective extreme environment.
Acidophiles, Alkalophiles, Thermophiles, Hyperthermophiles, Psychrophiles, Barophiles, Osmophiles, Piezophile, Polyextremophile, Xerophile. Bioinformatics
A) Introduction to Bioinformatics. Use of bioinformatics in major B) Major Bioinformatics Resources on Internet: National Centre for Biotechnology Information (NCBI).
The knowledge of various databases and bioinformatics tools
available at NCBI resource
ii.The major content of the NCBI databases.
iii.Purpose and applications in life sciences.
C) Protein data bank (PDB) and Nucleic acid sequence database D) The Basic Local Alignment Search Tool (BLAST), Nucleotide- nucleotide BLAST (blastn), Protein-protein BLAST (blastp), Specialized BLAST and FASTA Section II: Microbial Genetics
Basic concepts of microbial genetics
A) Structural organisation of Escherichia coli chromosome, folded B) Replication of DNA: DNA polymerase enzymes, mechanisms of replication and models – rolling circle and theta model C) Transcription, RNA polymerase enzyme, process and post D) Operon concept – Lac Operon, Nif gene regulation Bacterial mutations
D) Selection, detection & adaptation of mutants Genetic complementation
Genetic engineering and Protein engineering
A) Introduction, tools and techniques of genetic engineering C) Protein engineering – concept and application Techniques in molecular biology
B) DNA sequencing – Maxam and Gilbert’s method, Sanger’s method C) DNA finger printing – methods and applications (Paper VI)
Industrial Microbiology and Microbial Biochemistry
Section I: Industrial Microbiology
Food and diary Microbiology
A) Food as a substrate for microorganisms B) Food Fermentations – i) Idli ii) Bread C) Fermented dairy products: i) Cheese ii) curd iii) Butter iv) Kefir Industrial Microbiology
Industrial production of alcoholic beverages A) Grape wine – Definition, types, production of dry white table wine, Red table wine, Sparkling wines - champagne, California sherry, Microbial and non microbial spoilage of wines, Defects in wine. B) Beer – Definition, types, production of Lager beer and Ales Beer. Industrial production of -
D) Immunoactive products – toxoids, sera, vaccines E) rDNA products – Insulin, Interferon Unit IV Quality control in fermentation industry:
A) Test for sterility, pyrogenicity, allergy, carcinogenicity, toxicity for pharmaceutical and health care and food products filtration, cross flow filtration, flocculation, , whole broth processing solvent extraction, , concentration, Centrifugation, crystallization, distillation, adsorption elution, Precipitation and chromatography, Recent trends in fermentation Industry
A) Good manufacturing practices in fermentation industry B) Computer applications in fermentation technology Section II – Microbial Biochemistry
Enzyme kinetics and regulation (10)
A) Definition, properties, structure, specificity, mechanism of action (Lock and key model, induced fit hypothesis) B) Allosteric enzymes – Definition, models explaining mechanism of action D) Factors affecting catalysis efficiency of enzymes E) Enzyme kinetics – Derivation of Michaelis Menten equation, ii) End product repression – Tryptophan operon Extraction, purification and assay of enzymes
A) Extraction, homogenisation cell disruption and extraction of B) Purification of enzymes on the basis of iv) Adsorption characteristic differences. C) Immobilization of enzymes – method and applications Assimilation of :
i) Carbon ii) Nitrogen – N2 and NH3 (GOGAT) iii) Sulphur Bioenergetics:-Metabolic Pathways
E) Energy utilization in Bioluminescence. Biosynthesis of :
(Paper VII)
Agricultural and Environmental Microbiology
Section I: Agricultural Microbiology
Soil Microbiology
A) Introduction - Definition, approaches to soil microbiology, current C) Soil formation, structures and properties. D) Soil microorganisms, types and their role. E) Qualitative and quantitative methods to study soil fertility. F) Microbial interactions – symbiosis, commensalisms, amensalism, parasitism, predation. Role of microorganisms in
E) Interaction between micro organisms and metals Composting :
A) Compost production with reference to organic waste, availability of microorganisms, aeration, C:N:P ratio, moisture control, temperature, pH and time B) Biodegradation of Cellulose, Lignin, Pesticides and Hydrocarbons. Unit IV Plant pathology
A) Common symptoms produced by plant pathogens. B) Modes of transmission Plant diseases – oily spots on pomegranate – Xanthomonas oxyoposdis, white smut of sugarcane, soft rot of potato. Applications of Biotechnology in Agriculture
Section II: Environmental Microbiology
Air microbiology
A) Microorganisms in air – Launching, transport and deposition of aerosols, survival of microorganisms in air. B) Significance of microorganisms in air (extramural and intramural) C) Methods to study air borne microorganisms. Sampling, qualitative D) Bioaerosal control (ventilation, filtration, biocidal control, UV E) Sources, types, effects, control of air pollution.Depletion of ozone A) Marine microbiology Types of microorganisms in fresh and
marine water, estuaries, methods to study aquatic microorganisms.Chractrestics of marine environment, types of organisms and their role. B) Eutrophication:
Microbiology of potable water: Definition of potable water, standard
for potability, municipal water purification process, microorganisms as bioindicators of faecal pollution, routine bacteriological analysis of Environmental impact assessment:
A) Waste water assessment and management, types of wastes, chemical assessment, microflora, BOD, COD, treatment and disposal of waste water. B) Assessment and management of pollutants – toxic elements, their sources and effects and bioremediation (oris points, mercury, arsenic and radioactive substances) ii) Environmental legislations. iii) Environmental education and awareness A) Bio-leaching: Introduction, Microorganisms involved,
Biochemistry of microbial leaching, Commercial leaching – slope, heap, in situ leaching, Leaching of Copper and Uranium B) Oil recovery: Methods – primary, secondary, and microbialy enhanced, Biotechnological solution of oil recovery. C) Characteristics and treatment of wastes from different industries – paper and pulp,sugar and distillery, textile,dairy and antibiotic producing industries (Paper VIII)
Immunology and Medical Microbiology
Section I: Immunology
Complement system
Components of complement and their properties, Activation of complement – classical and alternate pathway, Biological effects of complement Major Histocompotibility complex
A. Organisation of MHC genes in man and mouse, classes of MHC
B .Adaptive immunology
a)Humeral (antibody) mediated – mechanisms, b)Cell mediated – cellular components, mechanism, cytokines A) Monoclonal antibodies – production (hybridoma technology). In
vivo and in vitro production. applications of monoclonal antibodies in diagnosis, research and treatment and research, B) Basis of antibody diversity: Basic concept of Immunoglobulin
A)Immunological tolerance
B)Autoimmunity – mechanism, types a) hemocytolytic b) organ
C) Hypersensitivity – types i) based on time required for
manifestation-Immediate and delayed. i)Based on pathogenesis-
anaphylactic reaction, atopy Catalytic and cytotoxic reaction. Immune
complex reaction-Arthus reaction, serum sickness. Delayed reaction. - tuberculin reaction- tuberculin test D)Transplantation Immunology – terminology, classes of
transplants, mechanisms of graft rejection, prevention of graft
rejection, HLA typing
Immunohematology: A)ABO blood group system, Rh blood group
B)National immunization programme: need and significance
C)Immunoprophalaxis – a.vaccines, Definition, types – conventional
(traditional),and new generation – a) DNA vaccines b) recombinant
B) Immunotherapy (passive immunization.)
Section II: Medical Microbiology
A) Pathogenicity of viruses, fungi and protozoa B) Hospital infection – factors contributing,,common types, diagnosis Microbial diseases (viral, fungi and bacteria) (Morphology, cultural and biochemical characters, antigenic structures, modes of transmission, pathogenesis, symptoms, laboratory diagnosis, prophylaxis and treatment of following diseases) Microbial diseases (viral and protozoan)
(Morphology, cultural and biochemical characters, antigenic structures, modes of transmission, pathogenesis, symptoms, laboratory diagnosis, prophylaxis and treatment of following diseases) A) Urinary tract infection – etiology, symptoms, diagnosis, B) Biomedical waste management – objective, contents, quality and types of wastes, waste treatment methods. C) Biological warfare –Use of biological agents - bacteria, viruses, or other disease-causing agents as biological weapons. Chemotherapy
B) Mode of action of following antimicrobial drugs: C) Penicillin, Bacitracin, Vancomycin, Isoniazid, Streptomycin, Chloramphenicol, Tetracycline, neomycin, erythromycin, rifampicin, quinolenes (cipro), Sulphonamide, Trimethoprim, Azidothymidine, Amphotericin,. D) Tests to guide chemotherapy a) diffusion tests b) broth dilution A) Reproduction of animal viruses – Adeno and Influenza.
B) Determination of number of viral population.
C) Oncogenesis – types of cancer, characteristics of cancerous cells,
D) Hypothesis of cancer:- Somatic mutation, viral gene, defective
Breed and Buchanan. Bergey’s Manual of Determinative Bacteriology, 9th Edition, 1974. General microbiology – Pawar and Daginawala Vol I and II Molecular Biology of Gene – J.D. Watson Genetics of bacteria and their viruses – William Hays Introduction of Bioinformatics – Affwood, T.K. Principles and techniques of Practical Biochemistry – K. Wilsons J.Walkar. Analytical Chemistry – Robert B. Dilts Chromatographic methods by Brathwaite and White Physiology and Biochemistry of Extremophiles by Charles Gerday and Nicolas Glansdorff (2007) Prescutt, Harley and Klein’s Microbiology, Willey Sherwood Woolverton, McGraw – Hill International Edition, (2008) Paper VI:
Outline of Biochemistry – Cohn and Stump Biological chemistry – Mahler and Cordes Enzyme structure and Mechanism – Alen fereht Principles of fermentation technology – Stanbury and Whitekar Pharmaceutical Microbiology – Huggo and Industrial Microbiology – Prescott and Dunn Food Microbiology – R.C. Dubey, D.K. Mahashwari Biochemistry – A problem approach by Wood, Hood and Weison Paper VII:
Microbial ecology – Fundamentals and applications - Atlas and Bartha Microbiology – Prescott and Harley, 5th edition Advances in Biotechnology – S.W. Jogdand. Textbook of Biotechnology – R.C. Dubey, Microbial dynamics and diversity – Desy Staley Biology of Microorganisms – Brock, Parker, Madigen, 9th edition Paper VIII:
Text book of Medical Microbiology – Ananthnarayan Review of Medical Microbioloyg – Jawetz et al Prescutt, Harley and Klein’s Microbiology, Willey Sherwood Woolverton, McGraw – Hill International Edition, (2008) Roitt M., (1984) Essentials of Immunology, P.G.Publishers Pvt. Ltd. New Delhi Roitt Evan, Brostoff J. Male D. (1993) Immunology 6th Ediction, Mosby and Co., London. Kuby J. (1996) Immunology Ed., 3 W.H. Freeman and Co. Davis and Dulbacco Medical Microbiology. Biochemistry of antibiotics – Franklin and Snow Medical laboratory technology – Ramnaik Sood Diagnostic Microbiology – Bailey’s and Scotts Immunology – a problem approach by Wood, Hood and Weison 16. Medical Bacteriology – Dey and Dey 17. G.P. Talwar (1983) Handbook of Immunology, Vikas Publishing Pvt. Ltd, New PRACTICAL COURSE
Practical I:
Isolation of DNA from bacteria by J. Mamar’s method Electrophoretic separation of protein (serum) & determination of phage cross infectivity. Determination of Optimum dose of U.V. by UV survival curve (comparative study of Escherichia coli and Staphylococcus aureus.) Isolation of Lac negative mutants of E.coli by visual detection method. Isolation of Streptomycin resistant mutant by gradient plate technique. Isolation of Vitamin B12 requiring (auxotrophic) mutants by replica plate technique. Diauxic growth curve of Escherichia coli (glucose and lactose). Browsing of National Centre for Biotechnology Information (NCBI) website.
Exploring protein sequence database (PDB) and GenBank and BLAST. Practical II:
i) DMC ii) Quantitative analysis of milk by SPC (using
Production of wine by using Jaggery medium by S. cerevisiae, examination of
pH, colour, taste.
Sterility testing of dry powder by direct inoculation on Soyabean casein digest medium. Immobilisation of enzyme by using Sodium alginate. Demonstration of crude recovery of amylase enzyme & Amylase assay. Estimation of Citric acid by titration method Practical III:
Isolation of microorganisms from soil. (Identification up to genus level) Rhizobium from root nodules.
Phosphate solubilising bacteria from soil.
Xantomonas from infected plant material
Estimation of available nitrogen from soil. Estimation of available phosphorous from soil (Stannous chloride method) Estimation of Calcium and Magnesium from soil (EDTA method) Determination of organic carbon contents of soil (Walkley and Black method) Setting up Winogradsky’s column and study of various types and groups. Study of Winogradsky’s column Study & analysis of fresh water flora. Microbiological analysis Drinking water: TPC, presumptive confirm and completed test. Determination of potability of water by MPN. Waste water analysis : physical (total solids), chemical (COD), biological (BOD) 17. Qualitative & Quantitative Analysis of Air Flora. & Determination of Practical IV:
Preparation of Tri sugar iron agar (TSI) medium slants and study of biochemical Determination of minimum inhibitory concentration (MIC).of penicillin 0n Staph aureus. Antibiotic sensitivity test (by disc diffusion method) Study of determination of effectiveness of antiseptic agents against selected organisms (Escherichia coli, Staphylococcus aureus, Bacillus), (antiseptic agents – tincture iodine, 3% H2O2, 70% alcohol, 5% chorine bleach) disc diffusion method A.Isolation of pathogen from clinical sample B.Isolation and identification of member of Enterobacteriaceae (Maximum Two) up to species level from biomedical waste (Gram staining, colony characterization, motility and biochemical tests – IMViC, H2S, oxidase, catalase, urea hydrolysis, gelatine hydrolysis, phenylalanine deaminase test, lysine decarboxylase test, sugar fermentation) Haematology – RBC count, WBC count, differential WBC count, Erythrocyte sedimentation rate (ESR) and demonstration of malarial parasite. a) Microscopic examination – pus cells, RBc, bacteria, crystals. b) Chemical examination – glucose (benedict’s method, protein (acetic acid), bile salt (sulphur method) ketone bodies (Rothera’s test) Study of synergistic action of antibiotics A) The practical examination will be conducted on four (4) consecutive days for not less than 6 hours on each day of the practical examination. B) Each candidate must produce a certificate from the head of the department in his/her college stating that he/she has completed in a satisfactory manner the practical course on the guidelines laid down from time to time by Academic council on the recommendation of Board of studies and has been recorded in his/her observation in the laboratory journal and written a report on each exercise performed. Every journal is to be checked and signed periodically by a member teaching staff and certified by the Head of the department at the end of the year. Candidate are to produce their journal at the time of practical examination Candidate have to visit (2) places of Microbiological interest(Pharmaceutical industry, Dairy, Research institutes etc) and submit the report of their visit at rhe time of examination. The report should be duly certified by the Head of the Department. Distribution of Marks for practical examination 1) One major experiment: 20 marks 2) Two minor experiment: 10 marks each 3) Journal : 5 marks. Total marks: Practical I Practical III : 45 Marks
Practical IV : 45 Marks
Tour Report : 20 Marks
Total Marks : 200

Practical wise distribution of marks
Practical – I
A) Major Experiments: (20)
i) Isolation of coli phages/ Mutants. OR
ii) Transformation of competent E.coli cells. OR
iii) Electrophoresis separation of DNA/Proteins.
B) Minor experiments: (20)
i) Diauxic growth / One step growth curve / Optimum dose of U.V by survival
ii) Isolation of chromosomal DNA/ Phage cross infertility /Browsing/ PDB/ Practical II
A) Major Experiments: (20)
i) Bioassay of Penicillin / Vitamin B12 OR ii) Amylase assay/OR iii) TLC of sugars / amino acidsOR iv) SPC of milk.OR Isolation of Lactic acid bacteria
B) Minor experiments: (20)
i) Direct microscopic count of milk / Phosphatase test of milk. & ii) Immobilization of enzymes.OR iii) Estimation of citric acid by titration/ alcohol by k2Cr2O7 Practical III
A) Major Experiments: (20)
i) Isolation of Azotobacter/Rhizobium/Xanthomonas OR
ii)BOD of sewage sample .OR iii).Estimation of nitrogen from soil. B) Minor experiments: (20)
i) COD of sewage sample ii) Estimation of calcium/ Magnesium/ Organic carbon of soil.
Practical – IV
A) Major Experiments: (20)
i) Isolation and identification of:
Pseudomonas aeruginosa/ Klebsiella pneumoniae/ Corynobacterium
ii) Isolation and identification of Enterobacteriaceae up to species level from
biomedical waste (Gram staining, colony characterization, motility and
biochemical tests – IMViC, H2S, oxidase, catalase, urea hydrolysis, gelatine
hydrolysis, phenylalanine deaminase test, lysine decarboxylase test, sugar
fermentation test)
B) Minor experiments: (20)
i) Determination of Antibiotic sensitivity to common pathogens.
ii) Serological tests.
iii) Hematological tests.
iv) Examination of urine sample.

List of the Minimum equipments and related requirements for B.Sc – III
1) Replica plating units for genetics experiments
2) Rotary shaker for fermentation experiments 12) Separate room for fine instruments of size 10’×15’ feet dimension 13) A separate culture room of atleast 10’×10’ feet dimension. 14) Electrophoresis assembly 20) Refrigerator :One 21) TLC UNIT : One 22) Winogradsky colomm


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