Laboratory Exercises in Microbiology,9780314872623

Laboratory Exercises in Microbiology

by ;
ISBN13: 9780314872623
ISBN10: 0314872620
Format: Paperback
Pub. Date: 1986-01-01
Publisher(s): Cengage Learning
List Price: $191.47

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Summary

This well-rounded lab manual provides the student with detailed insight into the theory behind each exercise, with careful instructions with helpful hints so that each procedure will work well. Accompanying explanations foster an understanding of why something is being done in the lab. The breadth of topics covered in this manner allow the manual to be tailored toward various types of students.

Table of Contents

Preface vii
I. MICROSCOPIC TECHNIQUES 1(22)
Use and care of the microscope
7(4)
Using the bright field light microscope
7(3)
Using the oil immersion lens directly
10(1)
Measuring cells using the microscope
11(2)
Observation of living microorganisms
13(3)
Making a wet mount
14(1)
Making a hanging drop mount
15(1)
Specialized light microscopes
16(7)
Dark field light microscope
16(2)
Phase contrast light microscope
18(2)
Fluorescence microscope
20(3)
II. CULTIVATION OF MICROORGANISMS 23(16)
Preparation of culture media
24(4)
Procedure for preparing a glucose minimal salts broth (a chemically-defined medium)
25(1)
Procedure for preparing a nutrient broth or agar (a chemically-complex medium)
26(1)
Procedure for autoclaving the medium
26(1)
Procedure for using the culture medium
26(1)
Procedure for preparing agar plates
27(1)
Microorganisms in the laboratory environment
28(2)
Pure culture techniques
30(9)
Procedure for making a streak plate
30(3)
Procedure for subculturing a pure culture into broth
33(1)
Procedure for subculturing a pure culture onto an agar slant
33(2)
Procedure for aseptic transfers using pipets
35(2)
Selective and differential media
37(2)
III. STAINING 39(24)
Positive and negative staining
41(3)
Procedure for positive staining
42(1)
Procedure for negative staining
43(1)
Differential staining
44(5)
Gram stain
44(3)
Acid-fast stain
47(2)
Structural staining
49(8)
Capsule stain
49(2)
Endospore stain
51(3)
Flagella stain
54(3)
Detection of inclusions
57(2)
Metachromatic (volutin) granules
57(1)
Poly-β-hydroxybutyric acid (PHB)
58(1)
Starch and glycogen
59(1)
Morphological unknown
59(4)
IV. ENUMERATION OF MICROORGANISMS 63(10)
Direct microscopic counts with a hemocytometer
64(2)
Viable counts
66(3)
Procedure for the pour plate method
67(1)
Procedure for the spread plate method
67(2)
Optical density measurements
69(4)
Procedure for determining population size using the pour plate method
70(1)
Procedure for determining the OD of the culture
70(3)
V. ENVIRONMENTAL FACTORS AFFECTING GROWTH 73(14)
Effect of temperature on microbial growth
75(1)
Effect of pH on microbial growth
76(1)
Effect of osmotic pressure on microbial growth
77(2)
Effect of NaCl on microorganisms
79(1)
Effect of sucrose on micro-organisms
79(1)
Effect of oxygen on microbial growth
79(6)
Growing organisms in an anaerobic gaspak jar
82(1)
Growing organisms in agar shake-cultures
83(1)
Growing organisms in a pyrogallic acid NaOH system
83(1)
Growing organisms in a candle jar
84(1)
Effect of light on microbial growth
85(2)
VI. CONTROL OF MICROBIAL GROWTH 87(10)
Physical methods of control
90(3)
The effectiveness of hand washing
90(1)
The effectiveness of heating
90(1)
The effectiveness of filtering
91(1)
The effectiveness of ultraviolet light
92(1)
Chemical methods of control
93(4)
Studying the factors that make an antimicrobial effective
93(1)
Determining the phenol coefficient
94(1)
Testing the bactericidal or bacteriostatic effect of antibiotics and drugs
95(1)
Determining the selective toxicity of antibiotics and drugs
95(2)
VII. METABOLIC ACTIVITIES OF MICROORGANISMS 97(26)
Hydrolysis of large extracellular molecules
98(6)
Hydrolysis of polysaccharides
98(2)
Hydrolysis of proteins
100(1)
Hydrolysis of nucleic acids (DNA)
101(1)
Hydrolysis of fats
102(2)
Fermentation of carbohydrates
104(3)
Fermentation of glucose, lactose, sucrose, maltose, and mannitol
104(2)
Methyl red test
106(1)
Voges-Proskauer test
106(1)
Respiration of carbohydrates
107(4)
Oxidase test
108(1)
Catalase test
109(1)
Nitrate reduction
109(2)
Utilization of amino acids
111(4)
Indole production
111(1)
Hydrogen sulfide production
111(1)
Lysine decarboxylase test
112(3)
Phenylalanine deamination
115(1)
Utilization of citrate, gelatin, and urea
115(3)
Citrate utilization test
116(1)
Gelatin hydrolysis
116(1)
Urea hydrolysis
117(1)
Identification of an unknown bacterium
118(5)
VIII. SURVEY OF EUKARYOTES AND CYANOBACTERIA 123(16)
The yeasts and molds
124(7)
Examination of cultures
124(2)
Coverslip preparation
126(3)
Sexual reproduction in Rhizopus
129(2)
The protozoa
131(3)
Examination of cultures
132(1)
Examination of pond water for protozoa
132(1)
Examination of prepared slides
133(1)
The algae
134(5)
Examination of algal cultures
135(2)
Examination of pond water for algae
137(2)
IX. THE VIRUSES 139(14)
Phase assay
143(1)
Bacteriophage isolation
144(1)
Plant virus isolation
145(1)
One-step growth curve
146(4)
Phage typing of Staphylococcus aureus
150(3)
X. GENETICS OF MICROORGANISMS 153(22)
Phenotypic and genotypic variation
154(5)
Illustrating phenotypic variation
154(1)
Isolating a mutant and illustrating genotypic variation
155(1)
Isolating streptomycin-resistant mutants (gradient plate technique)
155(2)
Replica plating
157(2)
Gene regulation
159(3)
Transformation
162(1)
Transduction
163(3)
Conjugation
166(5)
Mapping mutations and genes with F' and F- mutants
166(3)
Mapping genes by interrupting matings between Hfr and F- mutants
169(2)
Ames test
171(4)
XI. MICROBIAL ECOLOGY 175(20)
Population dynamics (growth curve)
176(1)
Microbial antagonism
177(3)
Competition
177(2)
Amensalism
179(1)
Microbial synergism
180(1)
Microbial populations in soil
180(4)
Isolating microbial populations
181(1)
Isolation of an endospore former
182(1)
Isolation of an antibiotic producer
183(1)
The nitrogen cycle
184(4)
Ammonification
185(1)
Nitrification
185(1)
Denitrification
186(1)
Nitrogen fixation
187(1)
The sulfur cycle
188(4)
The Winogradsky column
188(1)
Examination of photosynthetic sulfur bacteria
189(1)
Isolation of Thiobacillus
190(1)
Isolation of sulfate reducers
191(1)
Luminescent bacteria
192(1)
Periphytic bacteria
193(2)
XII. WATER SANITARY ANALYSIS 195(10)
Most probable number method for coliform analysis
196(2)
Membrane filter method for coliform analysis
198(2)
Noncoliform indicators of recreational water quality
200(2)
Isolation of Actinomyces from water and wastewater treatment plants
202(3)
XIII. FOOD MICROBIOLOGY 205(18)
Food spoilage
206(5)
Enumeration of food spoilage organisms
206(1)
Enumeration of proteolytic bacteria in food
207(1)
Enumeration of lipolytic bacteria
208(1)
Enumeration of molds and yeasts from fruit
208(3)
Food poisoning
211(1)
Evaluating the sanitary quality of foods
212(4)
Coliform analysis of foods
213(1)
Fecal streptococcal analysis of foods
214(2)
Examination of eating utensils and equipment
216(2)
Microbes and food production
218(5)
Sauerkraut production
218(3)
Vinegar production
221(1)
Microorganisms in sourdough production
221(2)
XIV. DAIRY MICROBIOLOGY 223(18)
Analysis of the sanitary quality of milk and dairy products
224(8)
Coliform analysis
224(1)
Dye reduction tests
224(1)
Direct microscopic counts
225(7)
Mastitis milk
232(3)
California mastitis test
232(1)
Modified Whiteside test
232(1)
Direct microscopic somatic cell count
233(1)
Cultural methods for mastitis milk
233(2)
Pasteurization of milk
235(2)
Microbes and the production of dairy foods
237(4)
Yogurt production
238(1)
Microorganisms and cheese production
238(3)
XV. AGRICULTURAL MICROBIOLOGY 241(8)
Plant pathogens
242(4)
Rusts and smuts
242(1)
Soft rot
242(2)
Crown gall
244(1)
Tobacco mosaic virus
245(1)
Mycorrhizae
246(1)
Bioinsecticides
246(1)
Examination of rumen fluid
247(2)
XVI. MEDICAL MICROBIOLOGY 249
Normal flora of the body
250
The oral cavity and cariogenic bacteria
250
Skin flora
251
Throat flora
251
Antibiotic sensitivity and the Kirby-Bauer procedure
252
ABO blood typing
255
Serological reactions
257
Bacterial agglutination test
257
The precipitin test
258
Immunofluorescence
262
The rapid plasma reagin (RPR) test for syphilis
264
Rapid multitest procedures for the identification of Enterobacteriaceae
266
The Enterotube II system
266
The API 20E system
268
APPENDICES
A. How to calculate dilutions
A1
B. Ranges and color changes of common pH indicators
B1
C. Organisms used in the laboratory manual
C1
D. Culture media
D1
E. Stains, indicators, and reagents
E1
Index I1

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