LABORATORY OUTLINE
NOTE: Due to the live cultures and complicated media preparation required for each lab, labs cannot be made up if missed.
A knee length lab coat must be brought to each week’s lab session. Lab coats cannot be left in the lab. It is recommended that you wash your lab coat after each lab.
Exercise # ( page)
WEEK # ACTIVITIES Benathen Brancaccio Taras/Muzio
1 Introduction, Safety Guidelines, Check-in iv- xviii v- vii
Culture Transfer Instruments, Techniques
& Isolation & Maintenance of Pure Cultures
(Aseptic Technique only) 5 (p. 25) 10 (p. 57)
MICROBIAL MORPHOLOGY
Smear Preparation and Simple Staining 6 (p. 33) 2 (p. 12)
Gram Stain 8 (p. 39) 4 (p. 25)
Bright Field Light Microscope 1 (p. 3) 1 (p. 3)
2 Gram Stain Practical
Acid-fast Staining (demonstration or class activity) 9 (p. 51) 5 (p. 31)
BACTERIAL ANATOMY
Endospore Staining 10 (p. 43) 7 (p. 41)
Capsule Staining (Graham & Evans procedure) 11 (p. 51) 6 (p. 36)
Bacterial Motility (demonstration or class activity) 12 (p. 53) 8 (p. 46)
The Effectiveness of Handwashing 25 (p. 117) 22 (p. 119)
3 MICROBIAL GROWTH
Determination of a Bacterial Growth Curve 16 (p. 75) 13 (p. 76)
(Two-hour method)
Environmental Factors Affecting Growth of
Microorganisms: Temperature, pH,
& Osmotic Pressure 20, 23-24 (p. 99) 15-17 (p. 89)
Cultivation of Anaerobic Bacteria (Broth &
Plate cultures- demonstration or class activity) 18 (p. 87) 18 (p. 101)
Exercise # ( page)
WEEK # ACTIVITIES Benathen Brancaccio Taras/Muzio
4 ISOLATION AND ENUMERATION OF MICROORGANISMS
Spread Plate Technique 14 (p. 63) 11 (p. 67)
Streak Plate Technique 14 (p. 63) 11 (p. 63)
Determination of Bacterial Numbers 15 (p. 71) 12 (p. 70)
Bacterial Count of a Food Product 30 (p. 151) 42 (p. 301)
Lethal Effects of Temperature (optional) 21 (p. 103)
5 CONTROL OF MICROBIAL GROWTH
Examination of Milk for Bacteria (Methylene Blue Reductase Test) 35 (p. 163)
Radiation (Ultraviolet Light) 45 (p. 249) 21 (p. 115)
The Effects of Chemical Agents on Bacteria I: Disinfectants 40 (p. 227) 19 (p. 105)
The Effects of Chemical Agents on Bacteria I:
Antimicrobial Agents (Kirby-Bauer method) 41 (p. 231) 20 (p. 109)
6 NORMAL FLORA, HOST DEFENSES,
AND UNKNOWN IDENTIFICATION
Nonspecific Host Defenses: Lysozyme (demonstration) 47 (p. 263) 44 (p. 315)
Isolation of
(Nose, Throat, or Skin) 48,49 (p. 267) 37 (p. 266)
Throat Culture Unknown Activity - Control and unknown
specimens inoculated
Proteins, Amino Acid & Enzymes V: catalase activity 36 (p. 184) 25 (p. 157)
Proteins, Amino Acid & Enzymes VI: coagulase
& DNase activity 49 (p. 271) 25 (p. 156,154)
7 TECHNIQUES TO ISOLATE AND IDENTIFY BACTERIA
Use of Selective and Differential Media 14 (p. 80)
Throat Culture Unknown Activity- Observe & Record results
Serology 45 (p. 324)
Exercise # ( page)
WEEK # ACTIVITIES Benathen Brancaccio Taras/Muzio
8 UNKNOWN IDENTIFICATION
Gastrointestinal (GI) Unknown Activity- Control & Unknown
specimens inoculated
Carbohydrate Fermentation 34 (p. 173) 23 (p. 128)
The IMViC Tests 37 (p. 195) 23,24 (p. 132,138)
Lysine Decarboxlyase Test 24 (p. 142)
9 GI Unknown Activity- Observe & Record results
Enterotube II System (demonstration) 39 (p. 215) 26 (p. 164)
10 Urinary Tract (UT) Unknown Activity- Control and Unknown
specimens inoculated
Hydrogen Sulfide Production using Kligler’s Iron Agar 37 (p. 195) 24 (p. 146)
IMViC Tests (Indole production only) 37 (p. 195) 24 (p. 139)
Gelatin Hydrolysis
Oxidase Test 36 (p. 184) 25 (p. 158)
Urease Activity 37 (p. 195) 24 (p. 143)
Phenylalanine Deamination 37 (p. 195) 24 (p. 140) Nitrate Reduction 36 (p. 184) 25 (p. 159)
ENVIRONMENTAL MICROBIOLOGY
Water Testing: COLILERT method 29 (p. 141) 30 (p. 205)
11 UT Unknown Activity- Observe & Record results
12 Final Laboratory Practical*
Check out**
*The final laboratory practical cannot be made up if missed. The point value for this practical (5%) will be deducted from your final grade.
**For check out, empty your drawer of all slides and media and place them in the discard bin. Clean empty drawer with microquat. Once the drawer has been cleaned, reusable materials can be returned to the drawer. Material in the slide discard bowls is placed in the discard bin. All paper trash is placed in the proper receptacle. Lastly, wash the laboratory bench top with microquat.