Laboratory Exercises in Microbiology

by Cano, Paul J.; Colome, Jaime S.

• ISBN13: 9780314872623
• ISBN10: 0314872620
• Format: Paperback
• Copyright: 1986-01-01
• Publisher: Cengage Learning

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