Laboratory Manual of Microbiology and Biotechnology [Paperback]

ISBN: 9789381714553

Edition: 1st Edition

Author : K.R. Aneja

Year: 2014

Pages: 424

Size: 21.5 x 28 cm

Publisher Name: Medtech

Rs. 595.00
Rs. 476.00

The intent of the manual is to provide basic and recent information and in depth methodology in a simple language about a wide spectrum of 144 exercises in a single tool essential for undergraduate and postgraduate courses in diverse areas of microbiology, biotechnology, biosciences, health-sciences, environmental science, agriculture, nutrition, pharmacy, medical laboratory technology, food processing units, and diagnostic laboratories. This clearly written, easy to use reference allows you to find what you need quickly.

Unique features of this manual are: each exercise provides useful background information; an in depth discussion of the principle involved; easy to follow the pattern of methodology with explanatory diagrams; expected results with interpretation profusely illustrated with coloured photographs to help students to perform practicals with their utmost ease in quest of achieving hundred per cent success; and the often asked questions at the time of viva-voce examination. This book is unique having experiments of multi-disciplinary nature and has a potential for a wide application. 

Unit I: Laboratory Safety Rules and Regulations

Unit II: Principle and Working of Basic Equipment

1.     Principle and Working of Laminar Air Flow Bench

2.     Principle and Working of an Autoclave

3.     Principle and Working of a Hot-Air Oven

4.     Principle and Working of a Bunsen Burner

5.     Principle and Working of an Incubator

6.     Principle and Working of a Colony Counter

7.     Principle and Working of Hemacytometer

8.     Principle and Working of a pH Meter

9.     Principle and Working of Water Bath

10.   Principle and Working of a Spectrophotometer

11.   Principle and Working of a Vacuum Filtration Assembly

Unit III: Microscopy and Basic Microbiological Techniques

12.   Bright-Field Microscopy

13.   Dark-Field Microscopy

14.   Fluorescence Microscopy

15.   Ocular Calibration and Microscopic Measurements

16.   Wet Mount Technique

17.    Hanging - Drop Preparation

Unit IV: Staining Techniques for Bacteria

18.   Preparation of Bacterial Smear

19.   Simple Staining

20.   Gram Staining

 21.   Acid-Fast Staining

 22.   Negative Staining

23.    Capsule Staining

24.    Endospore Staining

25.    Flagella Staining

Unit V: Biochemical Tests

26.    Durham Tube Sugar Fermentation Test

27.    Methyl Red Test

28.    Voges-Proskauer Test (V-P Test)

29.    Catalase Test

30.    Oxidase Test (Cytochrome Oxidase Test)

31.    Nitrate Reduction Test (Denitrification Test)

32.    Oxidation-Fermentation (O-F) Test

33.    Starch Hydrolysis Test

34.    asein Hydrolysis Test or (Caseinase Test or Skim Milk Utilisation Test)

35.    Gelatin Hydrolysis Test

36.    Urea Hydrolysis Test or Urease Test

37.    Indole Test (Tryptophan Hydrolysis Test)

38.    DNase Test (DNA Hydrolysis Test)

39.    Hydrogen Sulfide (H2S) Production Test

40.    Triple Sugar Iron (TSI) Agar Test

41.    Citrate Utilisation Test

42.    Decarboxylase Test

43.    Phenylalanine Deaminase (PAD) Test

44.    Litmus Milk Reactions

Unit VI: Culture Media Preparation

45.    Preparation of Nutrient Broth, Agar Plates, Stabs and Slants

46.    Preparation of Blood Agar

47.    Preparation of Chocolate Agar

48.    Preparation of MacConkey (MAC) Agar

49.    Preparation of Eosin Methylene Blue (EMB) Agar

Unit VII: Basic Microbiological Techniques for Culturing Bacteria

50.    Aseptic Transfer of Cultures

51.    Streak Plate Method

52.    Pour Plate Method

53.    Spread Plate Method

54.    Standard Plate Count

55.    Culture of Anaerobic Bacteria

Unit VIII: Methods used for Identifying unknown Bacterial Isolates

56.    Identification of Unknown Bacterial Isolates Using Conventional Methods

57.    Identification of Unknown Bacterial Isolates Using Multitest Systems Analytical Profile Index (API) System

58.    Identification of Unknown Bacterial Isolates Using Multitest Systems The Enterotube II System

 59     Identification of Unknown Bacterial Isolates Using Automated Methods

Unit IX: Study of Environmental Influences on Microbial Growth

60.     Bacterial Growth Cycle by Viable Counts or Plate Counts Technique

61.     Bacterial Growth Cycle by Measuring Turbidity (Growth Curve)

62.     Effect of Temperature on Growth

63.     Effect of pH on Growth

64.     Effect of Oxygen on Growth

Unit X: Evaluation of Antimicrobial Agents

65.     The Kirby-Bauer Method

66.     Epsilometer Test (E- Test)

67.     Broth Dilution Method

68.     Microdilution or Microtiter Method for MIC Testing

69.     Automated Antibiotic Susceptibility Test

70.     Evaluation of Ethanol as a Skin Antiseptic

71.     Evaluation of Disinfectants and Antiseptics: The Filter Paper Disk Method

72.     Effectiveness of Ultraviolet Light as a Disinfectant

Unit XI: Microbial Genetics and Molecular Techniques in Biotechnology

73.    Streptomycin-resistant Mutant Isolation by Replica Plating

74.    Antibiotic-resistant Mutant Isolation by Gradient-Plate Technique

75.    Mutant Production by UV and Isolation by Replica Plating

76.    Detection of Mutagens by Ames Test

77.    Genomic DNA Extraction from Bacteria

78.    Agarose Gel Electrophoresis for DNA Separation

79.    Isolation of Plasmid DNA from Bacteria

80.    Bacterial Conjugation

Unit XII: The Fungi: Molds, Yeasts and Mushrooms

81.    Cultivation and Identification of Molds

82.    Slide Culture of a Mold

83.    Cover Slip Culture Technique

84.    Cultivation of Fungi from Clinical Samples

85.    Cultivation of Yeasts

86.    Lactophenol Cotton Blue Wet Mount-Tease Mount Preparation

87.    Lactophenol Cotton Blue Wet Mount-Scotch Tape Preparation

88.    Potassium Hydroxide Wet Mount

89.    Potassium Hydroxide Lactophenol Cotton Blue Wet Mount

90.    India Ink Wet Mount

91.    Calcofluor White (CFW) Stain

92.    Morphology of a Mushroom

Unit XIII: Parasitic Protozoa and Helminths

93.   Study of Intestinal Protozoan Pathogens by Wet Mounts in Stool

94.   Acid-fast Staining of Stool Smears to Study Protozoan Parasites

95.   Leishman's Staining for Peripheral Blood Smears for Protozoan Parasites

 96    Saline and Iodine Wet Mount of Stool for Helminth Gastrointestinal Diseases

Unit XIV: Virology

97.    Enumeration of Bacteriophages by Plaque Assay Using the Agar OverlayTechnique

98     Bacteriophage Typing

99.    Culturing of Animal Viruses in Cell Cultures

100.  Isolation and Enumeration of Bacteriophages from Soil by Plaque Assay

101.  Study of Plant Viruses

Unit XV: Agricultural Microbiology

102.  Isolation of Plant Pathogenic Bacteria

103.  Examination of Plant Pathogenic Fungi in Moist Chamber

104.  Isolation of Plant Pathogenic Fungi

105.  Proving of Koch's Postulates for Plant Pathogenic Fungi

106.  Proving of Koch's Postulates for Plant Pathogenic Bacteria

107.  Isolation of Rhizobia from Root Nodules

109.  Nodulation Ability of Rhizobia

Unit XVI: Environmental (Soil, Air, Water) Microbiology

109.  Ubiquity of Microorganisms in the Environment

110.  Enumeration of Soil Bacteria by Plate Count Method

111   Enumeration of Soil Fungi

112.  Bacterial Interactions in Soil-Antagonism

113.  Bacterial Interactions in Soil-Commensalism

114.  Bacterial Interactions in Soil-Synergism

115.  Enumeration of Fungal and Bacterial Aerospora (Settle Plate Method)

116.  Bacterial Counts of Water by SPC

117.  Detection of Coliforms in Water (MPN Test)

118.  Determination of BOD of Water

Unit XVII: Applied Microbiology and Biotechnology

119. Bacterial Counts of Food

120. Detection of Foodborne Bacterial Pathogens

121. Methylene Blue Reduction Time Test

122. Microbial Spoilage of Canned Foods

123. Microbiology of Sauerkraut Production

124. Microbiology of Bread Production

125. Microbiology of Wine Production

126. Production of Ethyl Alcohol

127. Production of Penicillin

128. Production of Rhizobium as a Biofertiliser

Unit XVIII: Medical Microbiology and Immunology

 129. Proving of Koch's Postulates for Mice

 130. Normal Microbiota of the Skin

131.  Normal Microbiota of the Mouth

132.  Dental Caries Susceptibility Test

133.  Study of Sputum for TB

134.  Total Blood Cells Study

135.  Red Blood Cells Count in Blood

136.  White Blood Cells Count in Blood

137.  Differential White Blood Cell Count

138.  Collection and Preservation of Serum and Plasma

139.  Estimation of Haemoglobin in Blood by Haemoglobinometer

140.  ABO Blood Grouping by Hemagglutination Test

141.  Rh Blood Grouping

142.  Serological Typing of Bacterial Pathogens by Slide Agglutination Test

143.  Widal Test

144.  Enzyme-linked Immunosorbent Assay (ELISA)

Appendices

A.       Metric Measurements, Conversions, Math Tools, Generation Time and pH Equation

B.       Commonly used pH Indicators-pH Range, Colour Change and their Preparation

C.       Preparation of Solutions

D.       Preparation of Buffers

E.       Preparation of Biochemical Reagents

F.        Preparation of Stain's/Dyes

G.       Composition of Culture Media for Bacteria

H.       Composition of Culture Media for Fungi

I.         Sources for Microbial Cultures

J.        Methods for Collecting Clinical Specimens

K.       Classification of Prokaryotes (Bacteria and Archaea) based on Bergey's Manual of Systemic Bacteriology, 2nd            Edition (2004)

L.        Human Diseases and the Organisms that Cause them 

Dr. K.R. Aneja, former Professor and Chairman, Department of Microbiology, Kurukshetra University, Kurukshetra, is currently Chairman, Vaidyanath Research, Training and Diagnostic Centre (VRTDC), Vaidyanath Research and Healthcare Complex, Urban Estate Shopping
Centre, Sector-3, Kurukshetra (Haryana). He has a vast PG teaching and research experience of 38 years of which more than 11 years as Chairman of the Microbiology Department. Dr. Aneja has completed six major research projects sponsored by the UGC and Ministry of Environment and Forests, New Delhi and DFID, UK (on socio-economic impact and management of parthenium weed in India using indigenous fungal pathogens). Dr. Aneja and his research students have worked on fungal taxonomy, enzymology, microbial decomposition, biopesticides, and antimicrobial activity of fungi, plants and organic compounds against human and plant pathogens. He has published 155 research papers and reviews in International and National iournals of re ute.

He has attended many conferences and presented invited papers, and chaired technical sessions, and organised an International conference in 1997. He is a recipient of INSA-Royal Society Academic Exchange Fellowship and worked on protein profiling and PCR of thermophilic fungi in the University of Abertay, Dundee, Scotland. He is an active member of over 10 National and International societies and is a Fellow of five societies: Indian Botanical Society, Indian Phytopathological Society, Indian Mycological Society, Society for Biocontrol Advancement, and Mycological Society of India. He is the elected President of the Mycological Society of India for 2013. He is the elected Recorder of Plant Sciences Section of the Indian Science Congress (ISCA) for 2008 and 2009. He is an awardee of Rashtriya Gaurav Award and Best Citizens of India Award.

Dr. Aneja has authored/edited six books: "From Ethnomycology to Fungal Biotechnology" Kluwer Academic/Plenum Publishers, USA; "Glimpses in Plant Sciences" Pragati Prakashan, Meerut; "An Introduction to Mycology" Wiley Eastern Ltd., New Delhi; "A Textbook of Basic and Applied Microbiology", "Fungal Diversity and Biotechnology", and "Experiments in Microbiology, Plant Pathology and Biotechnology" New Age International Publishers. New Delhi.

Dr. Aneja is presently involved in environmental protection by applying the current strategies for controlling weeds so that the environmental pollution is minimised and humans get pesticide free food, and awakening the rural and urban people about the detrimental effects of pesticides. His research interests include: Fungal diversity. Environmental microbiology. Agricultural microbiology. Medical/Pharmaceutical/Diagnostic microbiology and Microbial biotechnology. 

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