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Micro5 > Lab Exam 2 > Flashcards

Lab Exam 2 Flashcards

1
Q

What are Koch postulates?

A
  1. Diseased organisms are diseased while healthy individuals are healthy
  2. Disease must be isolated and grown in the lab
  3. Inoculate a healthy host with the cultured disease to see if it causes the same disease
  4. reisolate disease and see if it is the same as original disease
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2
Q

What type of bacteria did we try to isolate from the yogurt?

A

Streptococcus thermophilus

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3
Q

Why did we only use 1 single colony of bacteria to inoculate in to the tube of milk?

A

To see if the “disease” yogurt will cause the same disease in the healthy (milk)

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4
Q

Why was it important to use only one type of colony to work with when viewing under the microscope?

A

To ensure we are looking at the same pathogen that is causing the disease

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5
Q

What is Koch postulates used for?

A

To determine the causative pathogen in animals, humans, and plants

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6
Q

Characteristics of healthy state milk

A
  • no smell, liquid, white w/ ph of 6.2
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7
Q

Characteristic of diseased yogurt

A
  • sour like smell, thick, white and ph of 4.0
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8
Q

What did the inoculation of milk do to it?

A

Caused it to become diseased - turned into yogurt

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9
Q

What characteristics (physical/chemical) cause a diseased and healthy individual different?

A

Color, smell, texture, ph

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10
Q

What could be done using modern technology to find out for sure that the same bacteria inoculated is the same bacteria causing the disease?

A

Sequence them!!

Take the diseased original and the inoculated disease and sequence the genome to see if the are the same pathogen

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11
Q

How did we know the reisolated bacterium was the same as the original bacterium causing disease?

A

They has the same color, consistency, ph and smell
(Same characteristics)

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12
Q

What are the 3 forms of physical media

A

Solid , semi solid and broth

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13
Q

What are the 2 types of chemical forms of media

A

Complex media & chemically defined media

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14
Q

Complex media

A

Media (nutrient broth & niftiest agar) that is enriched with yeast extract or beef extract

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15
Q

What is complex media used for?

A

Routine lab works to provide food for heterotrophic bacteria

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16
Q

Chemically defined media

A

More specific!!

Media with known amount of chemicals used

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17
Q

What is chemically defined media used for?

A

Grow organisms that require special requirements or exclude certain organisms

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18
Q

What are the 3 special purpose medias types.

A

Selective, differential and enriched media

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19
Q

Selective media

A

Suppress the growth of unwanted bacteria by encouraging growth of wanted bacteria

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20
Q

Differential media

A

Allows to distinguish between 2 different bacterial species growing

More physical!

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21
Q

Enriched media

A

Contain nutrients to Favor the growth of a certain bacteria

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22
Q

Example of selective media

A

MacConkey agar

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23
Q

Example of enriched media

A

Blood agar and chocolate agar

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24
Q

Differential media example

A

Blood agar

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25
Q

What are the 5 ways of sterilization?

A

Autoclaving, incineration, filteration, ultraviolet irradiation, chemical sterilization

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26
Q

Autoclaving

A

Expose steam at high temperatures and pressure in chamber to sterilize glassware and media

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27
Q

Incineration

A

Dry heat to destroy contaminated material

  • such as sterilizing loop/needle
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28
Q

Uv (ultraviolet) irradiation

A

Uv lights that damage dna

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29
Q

Chemical sterilization

A

Uses ethylene gas to sterilize heat sensitive items (plastic) by denaturing proteins

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30
Q

Filtration

A

Excludes microbes by being passed through a filter with pores

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31
Q

When is filtration used for?

A

To sterilize media that is inactivated by heat like antibiotics

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32
Q

Do disinfectants completely sterilize

TRUE or FALSE

A

No!!
It doesn’t kill them but rather reduces their growth

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33
Q

Disinfectants include

A

Microban, anti bacterial soap, bleach

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34
Q

What was the boiling water bath used to melt the agar in the media lab not sufficient enough to sterilize the media?

A

The temperature was 100 degrees Celsius which isn’t high enough to kill microbial life

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35
Q

Difference between sterilization and disinfection

A

Sterilization- removal of microbial life

Disinfection- reducing the amount of microbes present

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36
Q

What bacteria in the soil lab produces antibiotics

A

Streptomyces

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37
Q

What chemical produces the soil smell

A

Geosmin

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38
Q

What antibiotic was produced from streptomyces griseus

A

Streptomycin

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39
Q

What group does streptomyces belong to

A

Actinomycetes

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40
Q

What type of media was used in the soil lab? And why was it used?

A

Selective agar - AIA

— favors growth of actinomycetes and suppresses growth of molds

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41
Q

What are the characteristics of streptomyces colonies

A

Small, opaque, white, dry, powdery

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42
Q

Is streptomycin an narrow or broad spectrum of antibiotic?

A

Broad spectrum since it inhibits the growth of more then one broth culture

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43
Q

Total dilution =

A

Serial dilution x plating dilution

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44
Q

Plate dilution in soil lab

A

50ml on each plate -
1/100

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45
Q

What are the 4 other antibiotics produced by the genus streptomyces

A

Tetracycline, erythromycin, gentamicin, streptomycin

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46
Q

What are common food pathogens

A

E. coli, salmonella, shigella

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47
Q

How do you prevent food borne illness?

A

Properly washing vegetables, cooking meat all the way and using clean utensils

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48
Q

Do all food with a high CFU/g make you sick?

A

No!!
Some can contain good or healthy bacteria

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49
Q

Colony morphology

A

General appearance of bacterial colony ( shape, color, texture, size)

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50
Q

Why is colony morphology used?

A

To determine the identity of bacteria species and distinguish between differ t bacteria growing on the same plate

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51
Q

Why can bacteria colonies very in appearance?

A

Due to growing at different temperatures or growing on a different media type

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52
Q

What are the 8 ways to describe colony morphology

A

Configuration
Margin
Elevation
Size
Color
Texture
Optimal properties

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53
Q

Configuration describes

A

The shape

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54
Q

Margin describes

A

The edge or boarders

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55
Q

Elevation describes

A

The height of the colony

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56
Q

Examples of describing configuration

A

Round, irregular, rhizoid, filamentous

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57
Q

Examples of explaining the margin of colonies

A

Smooth, wavy, rhizoid, filamentous, curled, lobate

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58
Q

Examples of explains the elevation of bacterial colonies

A

Flat, raised, convex, droplike, fried egg

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59
Q

Examples of color to describe bacterial colonies

A

White, clear

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60
Q

Examples of sizes to describe colony morphology

A

Small, medium, large, x large

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61
Q

Examples of optimal properties to describe colony morphology

A

Translucent, opaque, shinny, dull, transparent

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62
Q

Examples that describe the surface texture

A

Smooth, rough, wrinkled, dry, moist and shiny

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63
Q

Chemical disinfectants are used to prevent/ reduce the growth of microbes on surfaces

TRUE or FALSE

A

True!!

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64
Q

Sterilization

A

Destruction of all microbial life enclosing endospores

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65
Q

Disinfection

A

Destruction of pathogens on inanimate objects

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66
Q

Antisepsis

A

Destruction of pathogens on living tissue

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67
Q

Sanitization

A

Lowering microbial counts on surface to safe level ( eating utensils)

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68
Q

Bactericidal

A

Substance that kills bacteria

69
Q

Sporicidal

A

Substance that kills spores

70
Q

What chemical agent is microban made out of?

A

Quat and alcohol

71
Q

What are the 4 chemical methods (types) used to control microbial growth?

A

Halogens, alcohol, quats, peroxide

72
Q

WhT are some of the most resistant microbes

A

Prions, endospores or bacteria & mycobacteria

73
Q

Why are mycobacteria resistant to disinfection?

A

They have a waxy cell wall that resists disinfectants

74
Q

What are more resistant to a disinfectant gram - or gram +?

A

Gram negative are more resistant due to having an outer membrane

75
Q

What are some of the least resistant microbes to disinfection?

A

Viruses and gram +

76
Q

Clorox wipes and antibacterial soap are what chemical agent

A

Quats

77
Q

Quats (quartering ammonium compounds) mechanism

A

Protein denature
Disruption of plasma membrane

78
Q

Bleach is an example of what chemical agent

A

Halogen

79
Q

Halogens mechanism of action

A

Alter cellular components
➡️ strong oxidizing agent

80
Q

Alcohol based hand sanitizer is an example of what chemical agent

A

Alcohol

81
Q

Alcohol’s mechanism of action

A

Denatures proteins
Disrupts plasma membrane

82
Q

Regular hand soap is an example of what chemical agent

A

Surfactant

83
Q

Surfactant mechanism of action

A

Washes away microbes ➡️ inserting into membranes

84
Q

Providence iodine is an example of what chemical agent

A

Halogen

85
Q

Hydrogen peroxide is an example of what chemical agent

A

Hydrogen peroxide

86
Q

Hydrogen peroxide mechanism of action

A

Alters cellular components by removing e- ( oxidizing agent)

87
Q

What are some ways to collect bacteria from a person?

A

Body surfaces - skin
Blood
Urine, stool or saliva

88
Q

How is bacteria collected from body surfaces?

A

Sterile swabs

89
Q

How is urine or stool collected?

A

Put in a sterile container

90
Q

How is blood collected for analysis?

A

Collected via sterile tube
- one aerobically and one anaerobically
- one w/ positive growth is streak plated

91
Q

What are three common types of agar media used to grow bacteria in a lab?

A

Blood agar, chocolate agar, macConkey agar

92
Q

Blood agar

A
  • enriched agar : containing animal blood
93
Q

How is blood agar both an enriched and a differential media?

A

It has the ability to distinguish pathogens by their ability to break down red blood cells and it’s made of animal blood

94
Q

Chocolate agar

A

Enriched media - used to culture certain medias such as heamophilus influenzae & neisseria gonorrhoeae

95
Q

How is chocolate agar an enriched he’d media

A

Lysed with blood cells that bacteria need in order to grow

96
Q

How is macConkey agar a selective and differential media?

A

Selective - inhibits growth of gram + so gram - can grow

Differential - distinguishes pathogens on their ability to ferment lactose

97
Q

What is added to macConkey agar to inhibit the growth of gram + bacteria?

A

Bile salts & crystal violet dye

98
Q

What is an example of lactose fermenting bacteria

A

E. coli

99
Q

What is blood agar used to culture

A

Pathogenic organisms to see if they can break down red blood cells

100
Q

Hymolysis

A

The ability for a bacteria to break down red blood cells

101
Q

Alpha hemolysis

A

Partial breakdown of red blood cells - looks yellow

102
Q

Beta hemolysis

A

Complete breakdown of red blood cells - looks green

103
Q

Gamma hemolysis

A

No breakdown of red blood cells - looks clear

➡️ example : staphylococcus epidermis

104
Q

5 steps in isolation and identification of bacteria from clinical samples

A
  1. Culture bacteria
  2. Obtain single colony
  3. Gram staining
  4. Metabolic test to complete plate bacterial Id
  5. Antibiotic sensitivity test
105
Q

Lactose fermenting bacteria produces acid that turns colonies pink

TRUE or FALSE

A

True

White colonies = no fermenting lactose

106
Q

Indigenous microbiota

A

Microbes that live on or in our bodies

107
Q

Do indigenous microbiota cause disease

A

No - not under regular circumstances

➡️ opportunistic pathogens can - if in the wrong place t the wrong time ( not where they belong)

108
Q

The Kirby Bauer method

A

Determines if bacteria is resistant to a particular drug

109
Q

Zones of inhibition

A

The area in which bacteria can not grow

110
Q

Broad spectrum of antibiotic

A

The antibiotic is effective against many types of microbes

111
Q

Narrow spectrum regarding antibiotics

A

Effective against only a couple microbes

112
Q

Antibiosis

A

Interaction between microbes where at least one is harmed

113
Q

What are 3 tests used to test antibiotics effectiveness?

A

E test
Vitek susceptibility test
Kirby Bauer method

114
Q

An example of a narrow spectrum antibiotic seen in lab

A

Penicillin, polymyxin B , Erythromycin

115
Q

Example of broad spectrum antibiotic seen in lab

A

Ampicillin, gentamicin, streptomycin and tetracycline

116
Q

If a bacteria is resistant to an antibiotic is there a zone of inhibition.

A

No - antibiotic did not stop growth

117
Q

Which antibiotics we tested are peptidoglycan synthesis inhibitors?

A

Ampicillin & penicillin

118
Q

What antibiotics did we test are protein synthesis inhibitors?

A

Streptomyces, tetracycline, chloramphenicol, Erythromycin & gentamicin

119
Q

What antibiotic targets the plasma membrane

A

Polymyxin B

120
Q

What antibiotic targets gram + bacteria

A

Erythromycin & penicillin

121
Q

Why are bacteria becoming resistant to many antibiotics

A

Antibiotics kill heathy and bad bacteria making antibiotics resistant bacteria more able to survive

122
Q

Kirby Bauer method

A

Test to determine the effectiveness of antibiotics

123
Q

E test

A

Plastic strip containing different concentrations of one type of antibiotic to find the minimum concentration it inhibits growth

124
Q

Vitek susceptibility test

A

A card containing wells with different antibiotics

125
Q

Sensitive means

A

Zone of inhibition is large

126
Q

Resistant means

A

Zone of inhibition is small

127
Q

What are the 5 things needed for the master mix - put into per tube

A

Template dna
Buffer
DNTPs (nucleotides)
DNA polymerase
RNA primer

128
Q

Taq DNA polymerase

A

Heat tolerant enzyme used to copy dna

129
Q

Primers

A

Short segment of dna that gives dna polymerase starting point

130
Q

Why does taq polymerase require a primer (rna)

A

It allows the adding of nucleotides to a new strand since taq polymerase can’t do it on its own

131
Q

What does pcr stand for?

A

Polymerase chain reaction

132
Q

What does a pcr do

A

Amplifies any dna sequence even from a small sample

133
Q

Amplify means

A

To make copies

134
Q

What is pcr tests used for

A

Infectious diseases, forensics and paternity tests

135
Q

What dna did you amplify in pcr lab?

A

Chromosome 16 from the cheek cell

136
Q

What was the site that was being tested in the pcr test?

A

Pv92

137
Q

3 steps to pcr test

A
  1. Denature (94c)
  2. Annealing (50-65c)
  3. Extension (75c)
138
Q

What is the alu sequence

A

An extra 300 base pairs w/ no known function

139
Q

Why is taq polymerase used in pcr?

A

It is stable at high temperatures so you don’t need to keep adding it

140
Q

If an individual is Alu +/+ where will the markers be after denaturing stage

A

One mark at the 941 base pair

141
Q

If an individual is +/- where will the markers be after the gel electrophoresis

A

One at the 941 base pair and one at the 641 base pair

142
Q

If an individual is Alu -/- where will the markers be after the gel electrophoresis

A

One at the 641 marker

143
Q

Electrophoresis

A

Process where dna fragments are separated according to size using electoral charge

144
Q

What about dna molecules allows you to separate them on gel

A

They have a negative phosphate backbone

145
Q

What is the purpose of adding loading dye in electrophoresis tank

A

To color the solution of sample

146
Q

What are the two components of the loading dye

A

Glycerol and bromophenol blue

147
Q

What was the role of glycerol in the loading dye ?

A

It helps the sample sink to the bottom of the well

148
Q

What is the role of putting bromophenol blue in the loading dye?

A

It allows us to see the dna on the gel

149
Q

What is the molecular marker purpose

A

It’s the template used to determine the size of our dna

150
Q

The purpose of the fast blast

A

To visualize dna

151
Q

What does ELISA stand for

A

Enzyme linked immunosorbet assay

152
Q

What is an antibody

A

A protein that acts against a specific antigen by attaching to it so the immune system can attack it

153
Q

What is an antigen

A

A toxic or foreign substance that causes an immune response in the body

154
Q

Each antibody binds to only a single matching antigen

TRUE or FALSE

A

True - they bind with specificity

155
Q

How are antibodies produced?

A

By B cells

156
Q

Where are antigens found

A

Surface of a pathogen

157
Q

What is tested to see if a patient has had a past infection

A

Antibody

158
Q

Where are antibodies found in

A

Plasma membrane

159
Q

What diseases are diagnosed with ELISA

A

Zika virus, hiv, Lyme disease

160
Q

What non disease is diagnosed with ELISA

A

Pregnancy

161
Q

If a test is not sensitive enough it might lead to what

A

A false negative

162
Q

Specificity of a test means

A

The test react to what it is supposed to

163
Q

If a test is not specific enough it might lead to what

A

False positive

164
Q

What was the role of horseradish peroxidase [HRP] in the ELISA experiment?

A

It linked to secondary antibody : produces a blue color when the substrate is added

165
Q

What does the intensity of the blue color indicate in ELISA test

A

How much antigen is present : darker the blue the more present

166
Q

Primary antibody

A

Binds to antigen

167
Q

Secondary antibody

A

Detects binding reaction of primary antibody

168
Q

Why did you assay positive and negative control samples in addition to your experimental samples?

A

The controls allowed is to know if it was done correctly- if contamination occurred or possible step skipped

Micro5 (14 decks)

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