Laboratory Manual of Microbiology

Description:
This laboratory manual of microbiology has been written to meet the needs of students taking microbiology as major or subsidiary subject. The intention is to provide the students with well organized, user-friendly tool to better enable them to understand laboratory aspects of microbiology as well as to hopefully make learning laboratory material and preparing for independent player of a given experiment. Each exercise provides step-by-step procedure to complete the assignment successfully and easily. The lab exercises are designed to give the student hands-on laboratory experience to better reinforce certain topics discussed in exercise. The glossary is included covering terms as well as basic, discipline-specific terminology from microbiology that will be helpful to its readers. The main contents of the manual are: Microbiology laboratory practices and safety rules, Basic laboratory techniques, Microscopy, Staining and motility techniques, Environmental microbiology, Microbiological culture techniques, Growth of lactose fermenting and non fermenting microbes, Medical microbiology, Environmental effect on bacterial growth, Application of microbiology, Microbiology of milk and Appendices. The academic level of the book is graduate, post graduate students, research workers, teachers and scientists dealing with basic and applied aspects of microbiology.

Contents:
1 Microbiology laboratory practices and safety rules
I BASIC LABORATORY TECHNIQUES
2. Media preparation
2.1. Sub culturing or picking off:
2.2. Broth cultures
3. Sterilization and disinfection
4. Inoculation and other aseptic procedures
4.1. Using a wire loop
4.2. Flaming procedure
4.3. Using a pipette
4.4. Flaming the neck of bottles and test tubes
5. Inoculation using a pipette
6. Pouring the media in Petri plate
7. Use of spreader
8. Spread plate technique
II MICROSCOPY
9. Use and care of microscope
10. Dark field microscopy
10.1. Bright field microscopy
10.2. Resolving power of microscope
10.3. Phase contrast field microscopy
10.4. Fluorescence microscopy
10.5. Electron microscopy
11. Ocular micrometery
III STAINING AND MOTILITY TECHNIQUES
12. Staining preparations
13. Smear making
14. Simple staining
15. Gram staining of bacteria
16. Capsule staining of bacteria
17. Capsule staining by negative, indirect or background staining
18. Acid fast (Ziehl-Neelsen) staining of bacteria
19. Endospore staining of bacteria
20. Dorner’s method of endospore staining
21. Bacterial flagella staining
22. Viability staining technique of bacteria
23. Lactophenol cotton blue staining of fungi
24. Staining of Vesicular Arbuscular Mycorrhiza (VAM)
25. Staining of actinomycetes culture
26. Hanging drop slide for bacterial motility
27. Motility Determination (Tube method)
28. Slide technique for protozoa motility
IV BIOCHEMICAL ACTIVITIES OF MICROBES
29. Urease activity of bacteria
30. Nitrate reduction test
31. Catalase test
32. Lipid hydrolysis test
33. Oxidase test
34. DNase production test
35. Starch hydrolysis test or amylase production test
36. Gelatin hydrolysis or production of gelatinase
37. IMVIC tests
37.1. Indole Test
37.2. Methyl Red and Voges-Proskauer Tests
37.3. Citrate Test
38. Hydrogen sulfide production and motility
39. TSI (Triple Sugar Iron) and KIA (Kligler's Iron Agar)
40. Casein hydrolysis
41. Estimation of phytohormone IAA production from bacteria
42. Estimation of ammonia production by bacteria
43. Solubilization of mineral phosphate by microbes
V ENVIRONMENTAL MICROBIOLOGY
44. Multiple tube fermentation tests
44.1. Presumptive coliform test
44.2. Confirmed coliform test
44.3. Completed coliform test
45. Counting of Ascaris eggs in raw or treated sewage water
46. Determination of dissolved oxygen (DO) of water
47. Determination of biological oxygen demand (BOD) of water
48. Determination of chemical oxygen demand (COD) of water
49. Detection of coliforms in water using membrane filtration technique
VI MICROBIOLOGAL CULTURE TECHNIQUES
50. Single Colony Isolation
51. Viable plate count or Pour plate or serial dilution agar technique
52. Counting of soil microorganisms
53. Isolation of Rhizobium from root nodules
54. Testing of nodulation ability by Rhizobia
55. Isolation of Azotobacter from soil
56. Carrier material for biofertilizers
57. Seed treatment with biofertilizers
58. Selection of bio-control agent
59. Isolation of pure culture and their preservation
60. Cultivation of anaerobic bacteria
VII GROWTH OF LACTOSE FERMENTING AND NON- FERMENTING MICROBES
61. Growth of bacteria using selective and differential media
62. EMB (Eosine methylene blue) agar for E. coli
63. MacConkey agar for enterobacterieaceae
64. Salmonella-Shigella (SS) agar for Salmonella and Shigella
65. XLD (Xylose Lysine Deoxycholate) agar for enterobacteriaceae
VIII MEDICAL MICROBIOLOGY
66. Micro-flora of upper respiratory tract
67. Enumeration of urine bacteria using pour plate technique
68. Use of urine dip slide for bacteria examination
69. Counting of bactiurea using calibrated loop method
70. Widal test: The tube agglutination test
71. Slide agglutination test for rapid diagnosis of pathogen
72. The Ouchterlony technique (double immuno-diffusion method)
73. Determination of Rh factor
IX ENVIRONMENTAL EFFECT ON BACTERIAL GROWTH
74. Effect of pH on bacterial growth
75. Effect of oxygen on the growth of microorganisms
76. Effect of temperature on growth of microorganisms
77. Effect of osmotic pressure on growth of microorganisms
78. Effect of radiation (Ultraviolet light) on growth of microorganism
X APPLICATION OF MICROBIOLOGY
79. Determination of viral plaque assay
80. The one step bacteriophage growth curve
81. Kirby-Bauer disk diffusion susceptibility test
82. Determination of phenol coefficient of a chemical or effect of disinfectant on bacteria
83. Determination of bacterial growth curve
84. Demonstration of Koch's postulates
85. Identifying Gram-negative enteric bacteria using API 20E system
86. Identifying Gram-negative, oxidase negative enteric bacteria using Enterotube II system
XI MICROBIOLOGY OF MILK
87. Examination of milk quality by methylene blue reduction test
88. Counting of number of bacteria in milk using Breed method