BIO 3722 / ES 3112 EXAM REVIEW SHEET

Please use this as a guide, while studying for the final exam. This should
supplement your notes and information in your lab manual. :o)

Safety
Recognize appropriate waste receptacles
->PUT NOTHING THAT IS CONTAMINATED INTO THE

REGULAR WASTEPAPER BASKETS. USED RUBBER GLOVES

ARE CONSIDERED TO BE CONTAMINATED.
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A step-wise procedure for cleaning broken contaminated glass.

-> IN THE CASE OF A SPILL OR ACCIDENT WITH ANY

CONTAMINATING MATERIAL (PRESENCE OF ANY

ORGANISMS) INFORM YOUR INSTRUCTORIMMEDIATELY,
AND THEN FLOOD THE CONTAMINATED AREA WITH
WESCODYNE AND COVER WITH PAPER TOWELS. AFTER
FIFTEEN MINUTES, PUT ON RUBBER GLOVES, PICK UP THE
TOWELS AND CONTAMINATED MATERIAL AND PLACE THEM
INTO A SMALL PLASTIC SACK. BROKEN GLASSWARE MUST
NOT BE HANDLED DIRECTLY BY HAND. CONTAMINATED
GLASS WILL BE PICKED UP USING A SMALL DUST PAN AND
BRUSH AND ALSO PLACED INTO THE PLASTIC SACK. THEN
PUT THE SACK INTO ONE OF THE LARGE RED (LAB 40)
OR LARGE WHITE (LAB 42) BIOHAZARD CONTAINERS. THE
DUST PAN AND BRUSH MUST BE RINSED WITH WATER AND
SPRAYED WITH LYSOL AFTER USE. CLEAN UP ALL
WESCODYNE WITH ADDITIONAL TOWELS. WASH YOUR
HANDS AS DESCRIBED IN PARAGRAPH FIVE.
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Proper hand washing technique

-> If you have a known spill of culture onto any skin surface wash the affected
area twice w/antimicrobial soap, rinsing with 70% ethanol btw washing. (enter
the lab, before starting lab, depart lab, remove gloves, sneeze or cough)

What PPE is required and when in this BSL II lab?

PASS
P- pull
A- aim

S- squeeze
s- sweep

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Section One
Pure culture is
Three physical states of media used in this lab.
Agars function, properties and purpose.
List transfer instruments used in this lab.

Ex. 1-4- Cx Transfer Techniques

Order of operationaseptic technique.

Why is it necessary to subculture microorganisms?

Ex. 1-5- Isolation Techniques

Four-way streak-plate: Rapid qualitative isolation method.

Zigzag method
Ex. 1-__: Sample Collection and Transport

Site, organism and destination.

Labeling
Ex. 1-6

Spread-plate: Quantitative isolation method.

Section 2: Controlling Microbial Growth
Chemical Methods vs. Physical Methods
Germicidal System Categories (pg 101)
Section 2: Microbial Growth
Ex. 2-1: Ubiquity of Microorganism

Terminology
Ex. 2-2: Colony Morphology

Agar Plates: 7 C.C.s (single isolated colony).

Ex. 2-4: Cultural Characteristics

Nutrient broth: precaution and observation

Ex. 2-5: Evaluation of Media

Biochemical families of life

Undefined vs. defined media

Fastidious organisms

Independent vs. dependent variable

Section 2: Controlling Microbial Growth
Chemical Methods vs. Physical Methods
Germicidal System Categories (pg 101)
Microbicidal effect vs. microbistatic effect
Chemical Methods: Antiseptics, Disinfectant, Chemotherapeutic agent
Range of microbicidal activity.
Ex. 2-6: Fluid Thioglycollate Medium
Ingredients of interest: sodium thioglycollate and resazurin
Why is a small amount of agar added to this medium?
Pre-analytical and post analytical precautions.
Aerotolerance categories (6)
Ex. 2-7: Anaerobic Jar
GasPak Anaerobic System BBLTM
Indicator tablet/strip reagent.

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EX. 2-8: The Effects of Temperature on Microbial Growth

Minimum, maximum and optimum growth temperature.

Psychrophiles, mesophiles and thermophiles (examples of each).

Prodigiosin

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Ex. 2-12: The Lethal Effects of Ultraviolet Light on Microbial Growth

Electromagnetic Spectrum
UV Radiation
Penetrability, distance vs. dosage, Thymine dimerization & Repair systems
(Ex. 8-2)
Ex. 2-11: Steam Sterilization

Parameters

Biological Indicators & Quality Control

Section Three- Microscopy and Staining
Ex. 3-1
Light microscopes vs. Electron microscopes
What type of microscope do we use in our lab?
Microscope components and their function
Function of immersion oil, purpose of Koehler illumination
The microscopes as precision instruments
Cell morphology

Depict or recognize shape and arrangement of microorganisms.

Monotrichous, lophotrichous, amphitrichous and peritrichous.

Ex. 3-4 Simple Stains
Chemical composition of a stain.
Acidic vs. Basic stains
Simple vs. Differential staining technique
Why are the air-drying and heat-fixing steps separate?
Ex. 3-5: Negative Staining
Practical advantages
Procedure and Observations.
Ex. 3-6 Gram Stain
Procedure and purpose of each reagent
Cell morphology
GP vs. GN cellular envelope
Ex. 3-7 Acid-Fast Stain
Procedure and purpose of each reagent
Ziehl-Nelson method (Z: hot) vs. Kinyoun method (K: cold)
Mycolic Acid
Genera of interest
Ex. 3-8- Capsule stain
Clinical significance
Procedure and purpose of each reagent
Cell morphology
Ex. 3-9- Spore stain
Sporogenesis vs. germination
Procedure and purpose of each reagent
Cell morphology
Ex. 3-11- Flagella stain

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Procedure and purpose of each reagent

Cell morphology
Section Four- Selective Media
When is its use appropriate?
Ex. 4-1 through 6
Special purpose, ingredients and relevant observations and conclusions.

Selective:

Differential:
Section Five- Differential Tests
Exoenzyme
Endoenzyme
Ex. 5-2,3, 4, 5, 6,7,11,12,13,14,18,19
Medium used for the test and ingredients that make the test possible.
Indicator and the end product of interest it detects.
Relevant Observation, Results and Conclusions (ORCs).
Presumptive Identification
False positives vs. false negative
1. Medium
2. Substrate
3. Enzyme
4. Mode of Action
5. End Product of Interest
6. Indicator/Reagent
7. Observations, Results, Conclusions
Ex. 5-4: Catalase Test
The great divider of Gram Positive Cocci!
Indicator and the end product of interest it detects.
Relevant ORCs.
False positives
Exp. 5-5: Oxidase Test
Tetramethyl-p-phenylenediamine(red)where is it and what is it doing?
Indicator and the end product of interest it detects.
Relevant ORCs.
False positives
Ex. 5-19: Triple Sugar Iron Agar
Medium used for the test and ingredients that make the test possible.
Indicator and the end product of interest it detects.
Relevant ORCs.
Please use the shorthand.
Ex. 5-21: BA
Streptococcal pathogens pathogens.
Media used for the test and ingredients that make the test possible.
Relevant Observation, Result and Conclusion (ORC)
Section Five- Differential Tests Ex. 5-10, 15, 20, 21, 22, & 23:
BEA, DNAse, Taxo A, CAMP, BA, and Coagulase
Streptococcal pathogens and Staphylococcal pathogens.
Media used for the test and ingredients that make the test possible.
Indicator and the end product of interest it detects.
Relevant Observation, Result and Conclusion (ORC)

Section Three- Microscopy and Staining

Ex. 3-3/11-1,2,3: Examination of Eukaryotic Microbes
The Helminths
Class, Transmission, Morphology, Adults, Infection, Treatment
Ex. T. solium: Cestoda, undercooked pork, circular egg, scolex w/ hooks and
suckers; monoecious, tapeworm infection, praziquantel.
Ex. E. vermicularis, Nematoda, fecal-oral route, bread loaf-like
eggs, dioecious, pinworm infection, mebendazole.
The Protozoa
Free-living: Phylum, mode of locomotion, morphology.

Ex. Vorticella: Ciliophora, cilia, contractile stalk.

Parasitic: Phylum, mode of locomotion, morphology, infection,
host(s), diagnositics.

Ex. Trypanosoma cruzi, Sarcomastigophora,

flagella, trypomastigote; Chagas Disease, Kissing bug (IH) & human (DH),
peripheral blood.

Ex. B. coli, Ciliophora, cilia, csyt vs. trophozoite, macro

vs. micronucleus; balantidiasis, human, stool sample.

The Fungi
Major characteristics: Nutrition, morphology, reproduction and
classification.
Organisms of interest: Rhizopus spp., Aspergillus spp., Penicillium
spp.,Candida albicans, Saccharomyces cerevisiae & Claviceps purpurea
Part Six: Quantitative Techniques
Ex. 6-1: Standard Plate Count (Viable Count)
Procedure, observations and results. All mental mathno calculators!
TV = vol. sample + vol. diluent
10-2, 1:100, .01, 1/100 J
OCD = CFU / FDF
Ex. 6-4: Differential Blood Cell Count
Blood: Liquid constituents and cellular constituents
Procedure: collection, smear prep, staining and diff (normal vs. abnormal)
Part Seven
Medical Microbiology: TO RAPIDLY ____________________,
___________________ & DETERMINE THE SUSCEPTIBILITY
OF ____________________________.
Ex. 7-1: Synder Test
Media used for the test and ingredients that make the test possible.
Indicator and the end product of interest it detects.
Relevant ORCs.
Dental-decay causing bacteria: _____________________________
Ex. 7-2: Antimicrobial Susceptibility Test (Kirby-Bauer Method)
Antibiotics vs. Synthetic Agents

Drugs of interest: Penicillin, ceftriaxone, ciprofloxacin, tetracycline,

chloramphenicol, rifampin, colistin, nalidixic acid, Augmentin (AmC),
Bactrim (SMX-TMP) and daptomycin. Mechanism of action, clinical
uses and adverse effects
Antibiotic sensitivity testing: Media, inoculation, incubation, observation
and interpretation.
Bergey's Manual of Determinative Bacterialogy: navigation,
interpretation, and unknown identification.
Terminology: Halophiles, baraophiles, acidophiles, neutrophiles and
alkaliphiles .
Please use this as a guide, while studying for the exam. This should
supplement your notes and the information in your lab manual. If you
have any questions, concerns or comments please call or e-mail
me. Study smart and get a good nights sleep before the exam. Im
rooting for all of you. :o)
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