Term
Exercise 1.1
Name the parts of a microscope and state the function of each. |
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Definition
Base - Support for microscope
Substage light - Illumination source located on base of microscope which shines light up through the condenser to the slide and object being viewed
Stage - platform that supports slides for viewing
Mechanical stage and control knobs - Apparatus which holds slides in place and which is used to adjust the (X and Y) position of the slide on the stage
Condenser - Substage lens that focuses the light on the specimen so that each of its points are evenly illuminated
Condenser knob - used to adjust the condenser up and down
Iris diaphragm lever - Opens and closes the condenser, thus regulating the amount of light shining through the condenser onto the object. Helps control contrast on the specimen
Coarse adjustment knob - Used with lower power objective lens to bring an object roughly in focus
Fine adjustment knob - used for fine focusing of all objective lenses
Head - Supports the ocular and objective lenses
Arm - Portion of microscope connecting the head and base
Ocular or eyepiece - Lens closest to the eye which magnifies the image produced by the objective lens
Rotating nosepiece (turret)- Structure on the head that holds all the objective lenses and which may be rotated to change from one lens to another
Objective lenses - lens closest to the specimen which magnifies the specimen
Scanning power - Generally about 4x magnification
Low power - generally about 10x magnification
High power - generally about 40x magnification
Oil immersion lens - Generally about 100x magnification and utilizes oil so that a more highly magnified and sharper image may be obtained
Rheostat - Used to vary the intensity of light |
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Term
Exercise 1.2
What are the total magnifications obtained with each objective lens? |
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Definition
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Term
Exercise 1.3
What is the resolution of this type of microscope? |
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Definition
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Term
Exercise 1.4
Why is blue light used when possible? |
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Definition
| Because blue light has the shortest wavelength of all visible light and resolution is inversely proportional to wavelength of light |
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Term
Exercise 1.5
What is the proper method to carry a microscope? |
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Definition
| With two hands. One hand holds the base and the other is on the arm of the microscope |
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Term
Exercise 1.6
What has to be used with every 100x objective lens? Why? |
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Definition
| Immersion oil because it increases the numerical aperture of the objective lens and prevents wide angles of light from being refracted away from the lens |
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Term
Exercise 1.7
This scope is parfocal. What does that mean? |
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Definition
| Once focused with a low power objective, the image remains always in focus for the other objective lenses, or very nearly in focus. This makes finding your specimen at high power much easier |
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Term
Exercise 1.8
Which objective lens has the smallest working distance? |
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Definition
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Term
Exercise 1.9
What are the three basic bacterial shapes? |
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Definition
| Bacilli (rod), cocci (spherical), spiral (helical) |
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Term
Exercise 1.10
What is always done to a microscope after using the 100x objective lens before it is put away in the cabinet? |
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Definition
| Clean off the oil from the lens |
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Term
Exercise 2.1
Will the media prepared from this powder be a liquid or solid media?
How do you know (besides looking at the name of the medium)? |
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Definition
| Solid. Look at the list of ingredients and look for the solidifying agent, agar |
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Term
Exercise 2.2
How should this medium be sterilized prior to its use? |
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Definition
| Autoclave at 15psi for 15 min, at 121 degrees C |
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Term
Exercise 2.3
To make up 500 ml of this media, how much of the following would you need?
a)water
b)Dehydrated media |
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Definition
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Term
Exercise 2.4
What percentage of casein will be in the final medium? |
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Definition
| 1.5% casein (15 grams in 1000 ml is like 1.5 in 100 ml, which would be 1.5%) |
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Term
Exercise 3.1
Be sure you know the basic aseptic microbiological transfer techniques you learned in Exercise 3 |
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Definition
| See Exercise 3 for specific details; review the proper techniques and sequences for transferring organisms from one type of media to another |
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Term
Exercise 3.2
If you wanted to isolate single colonies from a mixed culture, what type of culture would you use (e.g. streak plate, agar slant, broth)? |
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Definition
| Streak plate method (isolated colonies on an agar plate) |
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Term
Exercise 3.3
If you wanted to keep a pure culture of bacterium for a long time (weeks or a couple of months), what type of culture would you use (streak plate, agar slant, or broth)? |
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Definition
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Term
Exercise 4.1.1
What reagents did you use (and in what order) for each of the following stains?
a)Gram stain (Lab 4)
b)Schaffer-Fulton Endospore stain (Lab 8)
c)Capsule stain (Lab 11) |
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Definition
a) Crystal Violet, Iodine, Ethanol, Safranin
b)Malachite Green, Safranin
c)Capsule stain:
1)Mix equal amounts of India ink and water on glass slide, then add a loop of culture. Gently stir and spread mixture.
2)Allow smear to dry without heat fixation
3)Counterstain with methylene blue for 3 minutes
4)Rinse the slide very carefully and dry
5)Oil immersion. Blue cells w/ clear capsules on a black background |
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Term
Exercise 4.1.2
What is the function of each of the reagents for these stains |
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Definition
a)Gram Stain
Crystal violet - Primary stain
Iodine - Mordant (fixes stain)
Ethanol - Decolorizer
Safranin - Counterstain
b)Schaffer-Fulton Endospore Stain
Malachite Green - Primary stain
Safranin - Counterstain
c)Capsule stain
India ink - stain?
Methylene blue - counterstain |
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Term
Exercise 4.1.3
What procedure is used to apply and fix a bacterial smear for staining? |
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Definition
| Bacterial smears are prepared by placing a drop of broth culture on a clean microscope slide and spreading it in a circular motion. The smear is allowed to air dry and fixed by lightly passing it through the flame. If a smear is made from bacterial growth on an agar surface, a small drop of water is first placed on the slide and a very small amount of bacterial growth is emulsified in the drop to make a lightly turbid smear. It is also dried and fixed by flaming. |
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Term
Exercise 4.1.4
What about the bacterial cell is responsible for the characteristic response of a cell wall to the Gram stain? |
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Definition
| Gram Stain - The Gram stain reaction is dependent upon the nature of the cell wall of the bacterium. The Gram positive bacteria have a thick layer of peptidoglycan, which is able to hold the crystal violet-iodine complex within the cell wall when alcohol is applied. The Gram negative cell wall has only a thin layer of peptidoglycan with a lipid membrane around it. When the alcohol is applied it is not thick enough to hold the crystal violet-iodine complex. |
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Term
Exercise 4.2.1
Describe the arrangement of flagella on the bacterium shown in the figure below:
[image] |
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Definition
| Peritrichous - Many randomly all over (non-polar) |
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Term
Exercise 4.2.2
Can you name and describe the other types of flagellar arrangements? |
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Definition
Flagella Arrangements:
a)Monotrichous - Single flagella at one end (polar)
b)Lophotrichous - Many from one end (polar)
c)Amphitrichous - One or more at each end (bi-polar)
d) Peritrichous - Many randomly all over (non-polar) |
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Term
Exercise 4.3.1
What does the clear zone represent in this India ink preparation?
[image] |
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Definition
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Term
Exercise 4.3.2
[image]
What is the function? |
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Definition
| Protection against phagocytosis (inside the body) and dehydration (outside the body) |
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Term
Exercise 4.3.3
[image]
What two pneumonia-causing bacteria are noted for their large capsules? |
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Definition
| Streptococcus pneumonia and Klebsiella pneumoniae |
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Term
Exercise 4.3.4
[image]
Which one of the two would this one be? support your answer |
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Definition
| This one would be Klebsiella pneumoniae because Klebsiella is a bacillus and Streptococcus is a coccus |
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Term
Exercise 5.1.1
[image]
What fungus produced this special type of spore?
a)What is the function of fungal spores?
b)How does this function differ from the primary function of bacterial endospores? |
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Definition
Histoplasma capsulatum
a)Spores of fungi are primarily used by these microbes for reproduction. One mold can produce millions of spores, each of which can produce a new fungus colony. The spores also serve for dispersal and survival in dry situations.
b)The function of fungal spores contrasts with that of bacterial endospores whose essential function is for survival in unfavorable environments. Bacterial endospores are not used for reproduction/increases in number. One cell can only produce one endospore that germinate into one bacterial cell |
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Term
Exercise 5.1.2
[image]
Name the disease cause by this fungus |
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Definition
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Term
Exercise 5.1.3
[image]
At what temperature would the fungus have to grow to produce these spores? |
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Definition
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Term
Exercise 5.1.4
[image]
What is the phase of growth called? |
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Definition
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Term
Exercise 5.1.5
[image]
What is the specific name given to this spore type? |
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Definition
| Tuberculated chlamydospore |
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Term
Exercise 5.1.6
[image]
In what part of the country is this fungus found? |
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Definition
| U.S.A. (Eastern) Ohio and Mississippi River Valleys |
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Term
Exercise 5.1.7
[image]
How does one acquire this fungus infection? |
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Definition
| Inhalation of the infectious tuberculated chlamydospores |
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Term
Exercise 5.2.1
Look at the photo provided with this station ("Histoplasma capsulatum" page on fb) and notice the large, encapsulated yeast cells.
Name the following:
a)Disease
b)Fungus
c)Phase of growth |
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Definition
a)Cryptococcosis
b)Cryptococcus neoformans
c)Yeast phase |
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Term
Exercise 5.3
These are opportunistic yeast cells
[image]
1)What genus and species would be seen in this slide?
2)What disease could these cells cause?
3)How do yeast reproduce? |
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Definition
1)Candida albicans
2)Thrush, vaginitis, cutaneous candidiasis
3)Budding from mother cell |
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Term
Exercise 5.4.1
What type of organism is this? What is the name of this organism?
[image] |
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Definition
| This is a fungus known as Aspergillus |
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Term
Exercise 5.4.2
Is this organism a human pathogen?
[image] |
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Definition
| Aspergillus is an opportunistic pathogen of humans causing primarily respiratory tract infections (Aspergillosis) |
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Term
Exercise 5.4.3
What is the name of the toxin produced by this organism?
[image] |
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Definition
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Term
Exercise 5.4.4
What are some of the complications of this toxin?
[image] |
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Definition
| Can cause acute necrosis and cirrhosis of the liver leading to liver cancer |
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Term
Exercise 6.1.1
This is a tube of semisolid motility medium:
[image]
a)If there are two tubes displayed, which of the two tubes contains the motile bacteria?
b)If there is only one tube displayed, are the bacteria in this tube motile? How can you tell? |
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Definition
a) The motile bacteria would be in the tube where there is turbidity extending from the center inoculation stab. The one with the bacteria growing only down the center in the inoculation stab is the non-motile bacteria
b) No, one can tell because turbidity has not spread from the center stab line. Non-motile bacteria grow only along the stab and do not "swim" out from it to make the surrounding media turbid. |
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Term
Exercise 6.1.2
[image]
What other techniques can be used to determine bacterial motility? |
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Definition
| Flagella stain, hanging drop slide |
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Term
Exercise 6.1.3
[image]
By what means are most bacteria motile? |
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Definition
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Term
Exercise 6.1.4
[image]
What organelles are used by spirochetes for motility? |
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Definition
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Term
Exercise 6.2.1
What is this slide called?
[image] |
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Definition
| Concave or concavity slide |
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Term
Exercise 6.2.2
What is it used for?
[image] |
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Definition
| Motility observations using hanging drop preparations |
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Term
Exercise 6.2.3
What other techniques are used to demonstrate the same thing?
[image] |
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Definition
| Flagella stain, semisolid motility medium |
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Term
Exercise 7.1.1
Does ordinary hand washing remove all bacteria from the skin? Does a surgical scrub? |
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Definition
| Ordinary hand washing does not remove all bacteria from the skin neither does a surgical scrub. |
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Term
Exercise 7.1.2
Explain why hand washing is still done to prevent the transmission of disease |
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Definition
| To remove transient microbes (bacteria, viruses, fungi, etc.) that could be pathogenic and to reduce the contamination level of the skin |
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Term
Exercise 7.2.1
If surgical scrubbing does not remove all the microbes, why do a surgical scrub? |
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Definition
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Term
Exercise 7.2.2
What type of medium is blood agar? |
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Definition
| Enriched medium. It is also differential media because it allows one to determine the hemolytic patterns |
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Term
Exercise 7.2.3
Acquiring an infection from the rim of a drinking glass would be an example of transmission via a/an... |
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Definition
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Term
Exercise 8.1.1
What stain(s)/ staining procedure was used on this slide?
[image] |
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Definition
| Schaffer-Fulton Endospore stain |
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Term
Exercise 8.1.2
[image]
What genus of bacterium are these? What disease does this genus cause? |
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Definition
| Bacillus, anthrax (or Clostridium, gas gangrene, tetanus, botulism, pseudomembranous colitis) |
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Term
Exercise 8.1.3
What is the function of the green structures?
[image] |
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Definition
| The green structures are the bacterial endospores, used as a survival strategy during harsh conditions/lack of nutrients/lack of water |
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Term
Exercise 8.1.4
What special procedure has to be followed to stain the spores?
[image] |
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Definition
| Heating for 4 minutes in the presence of Malachite green stain |
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Term
Exercise 8.1.5
What dye in the stain was taken up by the following?
[image]
a)Vegetative cells
b)Endospore |
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Definition
Dye components take up by different parts:
a)Safranin
b)Malachite green |
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Term
Exercise 8.2.1
These bacteria (reddish bacilli) and tissue were taken from a diseased cow. Upon growth in the laboratory, endospores were formed
[image]
Name the disease that most likely killed this cow. |
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Definition
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Term
Exercise 8.2.2
These bacteria (reddish bacilli) and tissue were taken from a diseased cow. Upon growth in the laboratory, endospores were formed
[image]
What bacteria caused this disease? |
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Definition
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Term
Exercise 8.3.1
[image]
Name this apparatus |
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Definition
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Term
Exercise 8.3.2
[image]
What is its function? |
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Definition
| To provide anaerobic conditions for bacteria that require this type of atmospheric environment |
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Term
Exercise 8.3.3
What bacterial genus we studied this semester is grown in this type of apparatus?
[image] |
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Definition
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Term
Exercise 8.3.4
What diseases are caused by these bacteria (Clostridium)? |
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Definition
| Gas gangrene, tetanus, botulism |
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Term
Exercise 8.3.5
What other techniques are used to provide anaerobic conditions? |
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Definition
| Cooked meat medium, thioglycollate medium, anaerobic incubator |
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Term
Exercise 8.3.6
Can a candle jar be used? Why or why not? What is it used for? |
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Definition
| No, a candle jar cannot be used because the candle goes out before all the oxygen is burned up. Candle jars are used to provide an atmosphere with increased carbon dioxide for those bacteria such as Neisseria that grow better with it. |
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Term
Exercise 9.1.1
Review the Kinyoun Acid Fast Stain procedure |
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Definition
Kinyoun Acid Fast Stain procedure:
a)Flood air-dryed smear with Kinyoun Carbol Fuchsin for 5 min.
b)Rinse with water
c)Decolorize with Acidified alcohol
d)Rinse with water
e)Counterstain with Brilliant Green (or methylene blue) for 30 seconds
f)Rinse with water, blot dry, observe under oil-immersion |
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Term
Exercise 9.1.2
What color do Acid fast cells appear after this staining procedure? |
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Definition
| Acid fast cells (mycobacteria) should appear red |
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Term
Exercise 9.1.3
What is the name of the "waxy" lipid in the cell walls of Mycobacteria? |
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Definition
| The walls of mycobacteria contain Mycolic Acid |
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Term
Exercise 9.1.4
Name at least two diseases caused by Mycobacterium |
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Definition
Diseases:
a)Tuberculosis caused by Mycobacterium tuberculosis
b)Leprosy (Hansen's disease) caused by Mycobacterium leprae |
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Term
Exercise 10.1.1
Did regrigeration kill the bacteria in the experiment? How was this shown? |
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Definition
| No. When placed in the incubator after refrigeration the bacteria grew into colonies demonstrating that they were not killed but were merely inhibited from growing. |
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Term
Exercise 10.1.2
What term is used for a procedure that inhibits the growth of bacteria rather than killing them? |
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Definition
| The term bacteriostatic is used for inhibition of bacterial growth (microbiostatic would be used for any microbe, fungistatic for fungi, etc.) |
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Term
Exercise 10.1.3
Which of the two bacteria in this experiment was not killed by heating in the steam bath? Why? |
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Definition
| The Bacillus culture was able to grow after being exposed to 70C+ because it forms endospores that are resistant to temperatures this high. Vegetative cells cannot survive temperature this high |
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Term
Exercise 10.1.4
What term is used for a procedure that kills or destroys bacteria? |
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Definition
| The term bacteriocidal is used for procedures that destroy or kill bacteria (microbiocidal for microbes in general, virucidal for viruses, etc.) |
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Term
Exercise 10.1.5
Which of the bacteria were killed by autoclaving? |
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Definition
| Both bacteria were killed by autoclaving. Autoclaving destroys all microorganisms |
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Term
Exercise 10.1.6
What is the normal time and temperature (and pressure if applicable) of the following methods of microbial control?
a)Autoclave
b)Dry oven
c)Boiling |
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Definition
a)Autoclave - 121C, 15 mins, 15psi
b)Dry oven - 160-170C, 120-180 mins
c)Boiling - 100C, 15-20 mins |
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Term
Exercise 10.2.1
This is a photo of an antibiotic disk susceptibility test:
[image]
What is the formal name of this test? |
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Definition
| Kirby Bauer Disk susceptibility Test |
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Term
Exercise 10.2.2
This is a photo of an antibiotic disk susceptibility test
[image]
How would you determine if the bacteria cultured on these plates were susceptible to the antibiotics contained in the paper disks? |
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Definition
| Check the ones with the zones. To be accurate you would need a table of values to determine susceptible, intermediate, or resistant |
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Term
Exercise 10.2.3
This is a photo of an antibiotic disk susceptibility test
[image]
Why are some zones larger than others? Does a larger zone necessarily mean that the antibiotic is more effective? |
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Definition
| Because the size of the zone is inversely proportional to the size of the antibiotic molecule. Smaller molecules will diffuse faster and create larger zones of inhibition. Remember: One cannot compare the effectiveness of different antibiotics by their zones of inhibition in this test |
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Term
General Concepts 1
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Bacterial Capsules |
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Definition
Exercise 11:
Learned Gin's method of staining which consisted of India ink and Methylene blue stain |
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Term
General Concepts 2
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Bacterial motility |
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Definition
Chapter 6:
Learned how to do a hanging drop slide using a concavity slide |
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Term
General Concepts 3
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Resistance of bacteria to antibiotics |
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Definition
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Term
General Concepts 4
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Destruction of bacteria by autoclave and oven |
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Definition
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Term
General Concepts 5
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Effectiveness of hand washing on hand disinfection |
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Definition
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Term
General Concepts 6
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Safety of cafeteria eating utensils |
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Definition
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Term
General Concepts 7
General Concepts from Selected Laboratory Exercises
How were the following features of bacteria demonstrated in the laboratory?
Bacterial spores |
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Definition
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Term
Exercise 1.1.1
Referring to a microscope, state the function of the base |
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Definition
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Term
Exercise 1.1.2
Referring to a microscope, state the function of the substage light |
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Definition
| Illumination source located on base of microscope which shines light up through the condenser to the slide and object being viewed |
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Term
Exercise 1.1.3
Referring to a microscope, state the function of the stage |
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Definition
| Platform that supports the slide for viewing |
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Term
Exercise 1.1.4
Referring to a microscope, state the function of the mechanical stage and control knobs |
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Definition
| Apparatus which holds slides in place and which is used to adjust the (X and Y) position of the slide on the stage |
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Term
Exercise 1.1.5
Referring to a microscope, state the function of the condenser |
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Definition
| Substage lens that focuses the light on the specimen so that each of its points are evenly illuminated |
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Term
Exercise 1.1.6
Referring to a microscope, state the function of the condenser knob |
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Definition
| used to adjust the condenser up and down |
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Term
Exercise 1.1.7
Referring to a microscope, state the function of the iris diaphragm lever |
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Definition
| Opens and closes the condenser, thus regulating the amount of light shining through the condenser onto the object. Helps control contrast on the specimen |
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Term
Exercise 1.1.8
Referring to a microscope, state the function of the coarse adjustment knob |
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Definition
| Used with lower power objective lens to bring an object roughly in focus |
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Term
Exercise 1.1.9
Referring to a microscope, state the function of the fine adjustment knob |
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Definition
| used for fine focusing of all objective lenses |
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Term
Exercise 1.1.10
Referring to a microscope, state the function of the head |
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Definition
| Supports the ocular and objective lenses |
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Term
Exercise 1.1.11
Referring to a microscope, state the function of the arm |
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Definition
| portion of microscope connecting the head and base |
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Term
Exercise 1.1.12
Referring to a microscope, state the function of the ocular or eyepiece |
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Definition
| Lens closest to the eye which magnifies the image produced by the objective lens |
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Term
Exercise 1.1.13
Referring to a microscope, state the function of the rotating nosepiece (turret) |
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Definition
| Structure on the head that holds all the objective lenses and which may be rotated to change from one lens to the other |
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Term
Exercise 1.1.14
Referring to a microscope, state the function of the objective lens |
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Definition
| lens closest to the specimen which magnifies the specimen |
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Term
Exercise 1.1.15
Referring to a microscope, state the function of the scanning power |
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Definition
| Generally about 4x magnification |
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Term
Exercise 1.1.16
Referring to a microscope, state the function of the low power |
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Definition
| Generally about 10x magnification |
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Term
Exercise 1.1.17
Referring to a microscope, state the function of the high power |
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Definition
| Generally about 40x magnification |
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Term
Exercise 1.1.18
Referring to a microscope, state the function of the oil immersion lens |
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Definition
| Generally about 100x magnification and utilizes oil so that a more highly magnified and sharper image may be obtained |
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Term
Exercise 1.1.19
Referring to a microscope, state the function of the rheostat |
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Definition
| used to vary the intensity of light |
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