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Biology 15 - General Microbiology > Lab Exam 2 > Flashcards

Lab Exam 2 Flashcards

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1
Q

Pure Culture Techniques

What is a culture?
What is inoculate?
What is inoculum?

A
  • A culture is growing microbes in a laboratory setting.
  • Inoculate: add bacteria to a sample
  • Inoculum: Microbes in a growth medium
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2
Q

Pure Culture Techniques

What is the difference between a broth and agar?

A

Broth is a liquid media, while agar is a solid media.

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3
Q

Pure Culture Techniques

What are the types of agars that we use in class?

A
  • Flat top (slab)
  • Angled top (slant)
  • Petri dish (plates)
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4
Q

Pure Culture Techniques

What were the two tools used to inoculate new cultures?

A
  1. Cotton Swab
  2. Inoculating loop
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5
Q

Pure Culture Techniques

What does TSA and TSB stand for?

A
  • TSA = Tryptic Soy Agar
  • TSB = Tryptic Soy Broth

Tryptic contains amino acids.

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6
Q

Pure Culture Techniques

What is the difference between complex and defined media?

A

Complex media: We do not know the exact amount of ingredients.
Defined media: We do KNOW the exact the amount of ingredients.

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7
Q

Pure Culture Techniques

What are examples of nutrients that can be used in complex media?

A
  • Egg yolk
  • Yeast
  • Soy

TSA/TSB

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8
Q

Pure Culture Techniques

Is our standard media for our lab complex or defined?

A

Our media is complex

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9
Q

Pure Culture Techniques

What is the enrichment medium (even though we didn’t use them in our lab)?

A

They’re used to grow rare oragnisms. In order to do this we have to make growth conditions specific for the microbe type.

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10
Q

Pure Culture Techniques

What have been the two plating techniques introduced in the lab?

A
  1. Streak plate
  2. Lawn plate
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11
Q

Pure Culture Techniques

Which plating technique is used to produce pure, isolated colonies from a mixed sample?

A

Streak plate

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12
Q

Pure Culture Techniques

Why do we flame the loop between each zone when making a streak plate?

A

We want to reduce the number of bacterial cells. We would like to further dilute the sample.

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13
Q

Effect of Temperature on Growth

What are the organisms used for this exercise?

5 of them

A
  • Mycobacterium smegmatis
  • Escherichia coli
  • Staphylococcus aurues
  • Pseudomonas aeruginosa
  • Bacillus subtilis
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14
Q

Effect of Temperature on Growth

How did we apply our microorganism to the surface of this plate?

A

Used lawn plate technique/ cotton swab

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15
Q

Effect of Temperature on Growth

What are the temperatures that were chosen for this lab?

In celsius

A
  • 4 degrees celsius
  • 25 degrees celsius
  • 35 degrees celsius
  • 37 degrees celsius
  • 45 degrees celsius
  • 60 degrees celsius
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16
Q

Effect of Temperature on Growth

Which temperature in degrees celsius, is room temperature?

A

25 degrees celsius

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17
Q

Effect of Temperature on Growth

Which temperature in degrees celsius, is body temperature?

A

37 degrees celsius

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18
Q

Effect of Temperature on Growth

What temperature in degrees celsius, is refrigerator temperature?

A

4 degrees celsius

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19
Q

Effect of Temperature on Growth

What is the equation to convert celsius to fahrenheit?
What is the equation to convert fahrenheit to celsius?

A
  • 1.8(X celsius) + 32 = Fahrenheit or (X celsius multiplied by 9/5) + 32 = Fahrenheit
  • (X Fahrenheit - 32) x 5/9 = Celsius
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20
Q

Effect of Temperature on Growth

What optimal growth group did the temperatures grow best at?

A

35-45 degrees celsius

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21
Q

Effect of Temperature on Growth

What GENUS did not grow well at room temperature?

A

Mycobacterium

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22
Q

Effect of Temperature on Growth

Why didn’t the bacteria grow well at refrigerator temperature?

A

Because a refrigerator is at 4 degrees celsius. At this temp, it has a bacteriostasis effect on bacteria. This does not allow them to grow.

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23
Q

Evaluation of Antiseptics

Which organisms were used in this lab exercise?

The same 5 as the “Effects of Temperature”

A
  • Mycobacterium smegmatis
  • Escherichia coli
  • Staphylococcus aureus
  • Pseudomonas aeruginosa
  • Bacillus subtilis
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24
Q

Evaluation of Antiseptics

How did we apply our microorgansim to the surface of this plate?

A

Used lawn plate technique/ cotton swab

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25
Q

Evaluation of Antiseptics

What were the chemicals used in lab?

6 of them

A
  • Iodine
  • Hydrogen peroxide
  • Chlorhexidine
  • Isopropyl alcohol
  • Formaldehyde
  • Silver nitrate
26
Q

Evaluation of Antiseptics

How did we evaluate whether the antiseptic was effective or not?

A
  1. We can look at the data table to see how big or small the death zones are in mm.
  2. We can visually see the plate and see if there is a death zone or not

Death zone means that the anitseptic killed bacteria.

Big death zone = susceptible
Small death zone = resistant

27
Q

Evaluation of Antiseptics

Which organisms were the most and least resistant?

A
  • Most resistant: Escherichia coli
  • Least resistant: Mycobacterium smegmatis
28
Q

Evaluation of Antiseptics

Which chemcials were most and least effective?

A
  • Most effective: Formaldehyde
  • Least effective: Isopropyl alcohol
29
Q

Evaluation of Antiseptics

Each antiseptic has a chemical type, what are they?

A
  • Iodine = Halogen
  • Hydrogen peroxide = Oxidizing agent
  • Chlorohexidine = Biguanide
  • Isopropyl alcohol = Alcohol
  • Formaldehyde = Aldehyde
  • Silver nitrate = Heavy metal
30
Q

Evaluation of antibiotics

Which organisms were used in this lab exercise?

Same 5 as “Effects on temperature” and “Evaluation of antiseptics”

A
  • Mycobacterium smegmatis
  • Escherichia coli
  • Staphylococcus aureus
  • Pseudomonas aerugniosa
  • Bacillus subtillis
31
Q

Evaluation of antibiotics

How did we apply our microorganism to the surface of this plate?

A

Lawn plate technique/ cotton swab

32
Q

Evaluation of antibiotics

What were the six antibiotics used in this lab exercise?

A
  • Chloramphenicol
  • Cirprofloxacin
  • Gentamicin
  • Penicillin
  • Colistin
  • Bacitracin
33
Q

Evaluation of antibiotics

What are the basic modes of action for each antibiotic that was used?

6 of them

A
  • Chloramphenicol = Inhibiting protein synthesis
  • Cirprofloxacin = Nucleic acid inhibitor
  • Gentamicin = Inhibiting protein synthesis
  • Penicillin = Inhibiting cell wall synthesis
  • Colistin = Inhibiting cell wall synthesis
  • Bacitracin = Inhibiting cell wall synthesis
34
Q

Evaluation of antibiotics

How did we evaluate whether the antibiotic was effective or not?

Should be able to come to proper conclusions if given data table/agar plate

A
  1. We can look at data table to read the death zone in mm
  2. We can visually see the agar plate and spot the death zone
35
Q

Evaluation of antibiotics

Which antibiotic did not kill a few of the bacteria used in the experiment? How do we know that it did not kill? Does this selectively kill Gram + bacteria or Gram - bacteria?

A
  • Penicillin did not kill a few of the bacteria in the experiment. We know this because we don’t see a death zone for two of the bacteria (E. Coli & P. aeruginosa are gram -). This means that they selectively kill gram + bacteria.
  • Colistin did not kill a few of the bacteria in the experiment. We know this because we don’t see death zone for two of the bacteria (B. subtillis & S. aureus are gram +). This means they selectively kill gram - bacteria.
36
Q

Evaluation of antibiotics

Which organisims were most and least resistant?

A
  • Most resistant = Escherichia coli
  • Least resistant = Mycobacterium smegmatis
37
Q

Evaluation of antibiotics

Which antibiotics were most and least effective?

A
  • Most effective: Chloramphenicol
  • Least effective: Colistin
38
Q

Bacterial Transformation Lab

What bacteria did we use for this lab?

A

Escherichia Coli

39
Q

Bacterial Transformation Lab

What genes are found on the pGLO plasmid?

A
  • B-lactamase gene
  • GFP gene
  • araC gene
40
Q

Bacterial Transformation Lab

Which gene, when expressed, allows the colonies to glow?

A

GFP

41
Q

Bacterial Transformation Lab

What is the purpose of the LB in the plates?

A

To grow bacteria and provide nutrients

42
Q

Bacterial Transformation Lab

What inoculated plates or side of plates showed transformation?

A
  • LB/AMP+
  • LB/AMP/ARA

“ARA” because of arabinose

43
Q

Bacterial Transformation Lab

What is needed in the media for the colonies to glow?

A

Arabinose? I’m unsure of this answer.

44
Q

Bacterial Transformation Lab

What plate shows GFP expression?

A

LB/AMP/ARA

45
Q

Bacterial Transformation Lab

In the LB/AMP plate, what side (+ or -) should show growth?

A

In the LB/AMP plate, the + side should show growth.

Unsure about this answer, ask for help.

46
Q

Bacterial Transformation Lab

What is the name of the antibiotic resistance gene? What antibiotic is it resistant?

A
  • B-Lactamase
  • It is resistant against the amipicillin antibiotic.
47
Q

Bacterial Transformation Lab

What is transformation?

A

The bacterial cells ability to uptake naked DNA.

48
Q

Bacterial Transformation Lab

What are competent cells?

A

Competent cells are bacteria that can uptake naked DNA naturally. An example of this is Bacillus Subtilis.

Escherichia Coli is not naturally competent. Must be made competent.

49
Q

Bacterial Transformation Lab

How did we make the bacteria competent in the lab?

A

Cells were treated with calcium chloride and subjected to heat shock.

50
Q

Bacterial Transformation Lab

What is a plasmid?

A

Circular DNA found in bacteria cells.

51
Q

Bacterial Transformation Lab

What organism did GFP originate from?

A

GFP originates from marine jellyfish

52
Q

Selective and DIfferential Media

What is the definition of a selective media?

A

Inhibits certain bacteria while others survive.

53
Q

Selective and DIfferential Media

What is the definition of differential media?

A

Bacteria will grow but look different (color change).

54
Q

Selective and DIfferential Media

What were the three specific plates that were used in our lab?

A
  1. Eosin Methylene Blue
  2. Mannitol-Salt Agar
  3. MacConkey Agar
55
Q

Selective and DIfferential Media

Which plate used in lab this semester is not selective or differential?

This plate would not be part of the S&D experiment.

A

Hektoen Enteric plate

56
Q

Selective and DIfferential Media

Which organisms used in this exercise are Gram + ?

A

Staphylococcus auerus, Staphylococcus epidermidis

57
Q

Selective and DIfferential Media

Which organisms used in this exercise are Gram - ?

A

Escherichia coli, Enterobacter aerogenes, Proteus vulgaris

58
Q

Selective and DIfferential Media

What are the original colors of each type of plate before the experiment starts?

A
  1. Eosin Methylene Blue: Dark purple
  2. Mannitol-Salt Agar: Red
  3. MacConkey Agar: Berry
59
Q

Selective and DIfferential Media

How is each plate selective?

MSA, EMB, MAC

A
  • Mannitol Salt agar selects for Gram +, it is selective against Gram -
  • Eosin Methylene Blue and MacConkey agar selects for Gram -, it is selective against Gram +
60
Q

Selective and DIfferential Media

How is each plate differential?

MSA, EMB, MAC

A
  • Mannitol-Salt agar is differential based on mannitol fermentation.
  • Eosin Methylene Blue & MacConkey agar are differential based on lactose fermentation.
61
Q

Selective and DIfferential Media

Visually identify the results for the lab exam.

A

Biology 15 - General Microbiology (29 decks)

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