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From Flashcardlet > Micro Practical I > Flashcards

Micro Practical I Flashcards

1
Q

Binary Fission

A

single bacterial multiplies many tomes

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2
Q

Colony

A

a group of microbial cells that are visible to the naked eye

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3
Q

Genetically identical

A

cells that make up a colony

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4
Q

Medium

A

material that microbiologists use to grow organisms

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5
Q

Liquid Media

A
  • broth
  • placed in a test tube
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6
Q

Agar

A
  • solid media
  • test tube or petri plate
  • “nutrient” agar
  • melts at 100oC, solidifies at 42oC
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7
Q

Aseptic Technique

A
  1. protects culture from being contaminated
  2. protects you & env’t from being contaminated with microbes
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8
Q

Pure Culture

A
  • only one species of microbes, colonies look identical
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9
Q

Mixed Cultures

A
  • different colonies.
  • must sterilize innoculation needles, for sterilization
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10
Q

Colony Morphology

A
  • Size
  • Pigment
  • Optical Characteristics: translucent, dull, glossy
  • Form
  • Elevation
  • Texture
  • Consistency
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11
Q

Form/elevation

A
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12
Q

Ubiquity Testing

A
  • Bottom of shoe and finger
  • green, yellow
  • orange
  • white
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13
Q

Can you tell a mixed culture from a pure culture?

A

can never know for sure

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14
Q

How many different genetic types of cells are there in a colony?

A

one

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15
Q

why are cells in a colony considered a clone?

A
  • bacteria propogate by binary fission, all are genetic clones.
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16
Q

Resolving Power

A
  • the smallest distance between 2 objects that can be clearily seen
  • RP= wavelength/2 x NA
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17
Q

Purpose of Oil immersion

A

prevents refraction of light

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18
Q

Smallest object that can be seen with your microscope

A

550nm/2x1.25= 220nm

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19
Q

Three types of bacterial staining techniques

A
  1. simple
  2. differential
  3. structural
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20
Q

Differential

A
  • distinguishing between groups
  • Ex: Gram Stain, Acid Fast
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21
Q

Simple Stain

A
  • see overall structure but not cell wall, cytoplasm, flagella
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22
Q

Structural Stain

A
  • if bacteria possess important structures such as
    • flagella, capsules, and endospores
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23
Q

Gram negative Cell Wall

A
  • has an outer membrane
  • smaller peptiglycan
  • periplasmic space
  • lipopolysaccharides (LPS)
  • Pink
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24
Q

Gram Stain Steps

A

Crystal–> 60 sec—>H2O—>Gram Iodine—>60 sec—>H2O—>Alocohol—>10-15 sec—>H2O—> Safranin–>60 sec

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25
Q

Mordant

A
  • Iodine
  • causes dye to form large crystals
    • get trapped in Gram positive in pepti layer
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26
Q

Safranin

A
  • in order to make cells visible once again
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27
Q

Gram stain + Reliable results

A
  • should be younger (24-48 hours old)
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28
Q

Staphyloccus Epidermis Positive Gram Stain

A
  • purple
  • cocci
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29
Q

Escherichia Coli Negative Gram Stain

A
  • pink
  • rods
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30
Q

Acid Fast Test

A
  • Ziehl-Neelsen stain,
  • acid fast bacilli are stained bright red and stand out clearly against a blue background
  • Mycobacterium
  • waxy, similar to gram +
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31
Q

Capsules

A
  • polysaccharides
  • presence of capsule= ability to cause disease
  • act as adhesins to attach to host tissue
  • cannot use HEAT
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32
Q

Kinyoun

A
  • cold acid fast test
    1. Phenol
    2. Acid Alcohol
    3. Methylene blue, counterstain
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33
Q

Flagella

A
  • gently
  • no heat, air dry
  • below RP of microscope 15n diameter
  • move away from toxic compounds
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34
Q

Endo spores

A
  • vegetative cells stain RED
  • Spores stain GREEN
  • resistant to heat, radiation, disinfectant,and desicattion
  • Does not grow or divide, resisitent to temperature
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35
Q

Why is it necessary to heat the slide continusouly during endospore process?

A
  • cooking malachite green into the endospore wall. very resistant to staining
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36
Q

Why is it tricky to stain flagella

A
  • are extremely brittle
  • will break if subjected to mechanical forces
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37
Q

How does bacteria benefit from having a capsule?

A
  • helps to protect bacteria against phagocytosis
  • contain water which protects bacteria against desiccation.
  • exclude bacterial viruses and hydrophobic toxic materials such as detergents.
  • found most commonly among gram-negative bacteria, such as E.coli
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38
Q

Why do microbiologists stain bacteria?

A
  • makes microorganisms easier to see
  • because it adds color and contrast
  • not alive so not dangerous
  • not a source of contamination
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39
Q

What is the purpose of heat fixing bacterial smears before staining?

A
  • make it adhere to the slide
  • make organism accepts stains
  • kill organism
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40
Q

Why do acid-fast bacteria retain the carbolfuschin stain?

A
  • because the dye is more soluble in the cell walls lipids than in the acid alcohol.
  • retain the initial color of the first dye because they are able to resist the destaining by acid alcohol.
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41
Q

Streak Plate

A
  • streaking in quadrants to eventually dilute
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42
Q

Spread Plate

A
  • 1:10, 1:100
  • 30-300 manageable
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43
Q

Isolation Techniques

A
  • Spread Plate
  • Streak Plate
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44
Q

Dilution Factor

A
  • # of bacterial cells = # of colonies on the plat X dilution factor/ ML of diultion plated
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45
Q

Sacromyces Cervesi

A
  • beer yeast
  • single-celled fungi
  • multiply by budding, or
  • by division (fission)
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46
Q

Isolation Technique Class Average

A

564,500 cfu/mL

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47
Q

Environmental Factors that influence the growth rate (x6)

A
  1. oxygen
  2. temperature
  3. osmotic pressure
  4. pH
  5. UV
  6. CO2
48
Q

Thermophiles

A

grow best at 40oC to 90oC

49
Q

Mesophiles

A
  • body temperature
  • 20oC to 40oC
50
Q

Psychrophiles

A
  • cold temps
  • 0oC to 20oC
51
Q

Osmotic Pressure

A
  • move from high concentration to low concentration
52
Q

halophiles

A

love salt

53
Q

pH

A

acidic = high concentration of H+

54
Q

UV

A
  • causes DNA mutations
  • or Microbicidal (kill microbes), over long period
55
Q

Effects of Temperature on P.Fluorescens

A
  • 4oC phile
  • **psychrophile **
56
Q

Effects of Temperature on E.Coli

A
  • 37oC phile
  • 55oC tolerant
  • mesophile
57
Q

Effects of temperature on B.Stearothermophilus

A
  • 37oC tolerant
  • 55oC phile
  • thermophile
58
Q

Effects of Osmotic Pressure on P.Fluorescens

A
  • 0.5% NaCL phile
  • halophilic
59
Q

Effects of Osmotic Pressure on E.Coli

A
  • halotolerant
60
Q

Effects of Osmotic Pressure on S. Epidermidis

A
  • halophile
61
Q

Effects of pH on P.Fluorescens

A
  • Neutra-tolerant
  • Alkala-phile
62
Q

Effects of pH on E.Coli

A
  • acidophile
  • neutratolerant
63
Q

effects of pH on Se.Epidermidis

A
  • neutrophilic
  • alkalitolerant
64
Q

Effects of UV radiation on E.Coli

A

when UV increases exposure, bacteria drops

65
Q

Why are endospore forming bacteria considered thermoduric and not thermophilic

A
  • use spore coat as a defense
66
Q

Hypotonic why does water rush into the cell and cause to swell

A
  • water fills the lower concentration
67
Q

What protects most bacteria from swelling up and bursting in hyptonic solutions?

A
  • cell wall protects microbes growing in natural hypotonic conditions
  • osmotic lysis
68
Q

What type of organism prefers to grow in hyper-tonic solutions?

A
  • 5% and 15% NaCL
69
Q

How many more time acidic is lemon juice pH 2 to tomatoes pH 4

A

103

70
Q

Why would your UV radiation experiment not work if you forgot to remove the lide of the plate while irradiating?

A

because uv light cannot get through plastic

71
Q

Aerobes

A

use oxygen for metabolism

72
Q

Anaerobes

A
  • can grow in the absense of oxygen
73
Q

Facultative Anaerobes

A
  • capable of growing in both oxygen and no oxgen
  • grow better WITH oxygen
74
Q

Obligate Anaerobes

A
  • killed in the presence of oxygen
75
Q

Aerotolerant Anaerobes

A
  • always perform anaerobic metabolism
  • NOT killed in the presence of oxygen
76
Q

Microaerophiles

A
  • low concentrations of oxygen
  • 3-5% CO2
77
Q

Neutralizatino of Superoxide Dismutase

A

202 + 2H –> O2 + H2O2

78
Q

Neutralization of Catalase

A

2H2O2 –> 2H2O + O2

79
Q

Gram Negative

A
  • pink stained
  • rod shaped
80
Q

Endospore Staining

A
  • red stained rods
  • green stained spheres
81
Q

Flagella Stain

A
  • Petrichious Flagella
  • Purple Stained Rods
  • multiple purple lash-like appendages
82
Q

Acid Fast Stain

A
  • Mixed Culture:
  • Blue: Staphylococcus epidermidis, spherical
  • Red: Mycobacterium smegmatis- rod shaped
83
Q

IMViC Tests

A
  1. Indole Production
  2. Methyl-Red
  3. Voges Praskauer
  4. Citrate Utilization
    1. differentiate e.coli
84
Q

Tryptophanase

A
  • removes both the R-indole and amine groups
  • produces pyruvate, ammonia, & indole
  • determined by Kovac’s reagant
85
Q

Kovac’s Reagant

A
  • determines Tryptophan digestion
  • detect production of indole
  • Positive test= RED
  • E.coli +, E.Aerogene _
86
Q

Methyl Red

A
  • Mixed Acidic Pathway
  • inoculated broth with organism
  • Red at acidic pH
  • E.coli +, E.Aerogenes -
87
Q

Voges Proskauer

A
  • Reagant is Barritt’s A (alpha natphol) and B(potassium hydroxide.
  • Pink Burgundy= positive test
  • Butylene Glycol Pathway (no pH)
  • E.coli - , E.Aerogenes +
88
Q

Citrate Utilzation

A
  • Citrate sole carbon source
  • Ammonium dihydrogen phosphate sole Nitrogen source.
  • pH indicator Bromothymol Blue= Reagant
  • Positive= blue
  • E.Coli= Negative. E.aerogenes=Positive
89
Q

Oxygen Radicals

A
  • oxygen molecules have too man or too few electrons
90
Q

ROS

A
  • Reactive oxygen species.
  • by product of oxygen metabolism
  • harm macromolecules
  • Example ; Hydrogen Peroxide
  • Must detox converting to water and oxygen
91
Q

Cytochrome Oxidase

A
  • reduce oxygen at end of electron transport chain
  • differentiate between neisseriaceae, pseudomonadace, enterobacteria
92
Q

Selective Medium

A
  • inhibit growth of certain organisms
93
Q

Differential Medium

A
  • Allow all organisms to grow, but they have components that allow to distinguish between certain groups
94
Q

Levine EMB

A
  • agar
  • selective & differential
  • contains Lactose
  • differentiating from e.coli and e.aerogenes
95
Q

Luisetti’s Medium

A
  • differential
  • detects fluorescence of pseudomonas.
96
Q

Brownian motion

A
  • random motion of molecules in a liquid
97
Q

Why grow faster under aerobic vs anaerobic?

A
  • because it produces more ATP
98
Q

Why did we put resazurin in thioglycolate medium?

A
  • dye indicator of the presence of oxygen
  • Red in the presence of oxygen and colorless under anaerobic conditions
99
Q

Of the 3 fermentation products, which one did we NOt test for carb fermentation?

A
  • Microaerophilic
100
Q

2 possible arrangements of flagella

A
  1. Petrichous- ransomly dispersed
  2. Polar- located at the ends
101
Q

Motility Medium/Result

A
  • small amt of agar, semi-solid
  • +, seen as turbidity
102
Q

What is the substrate catabolized to produce indole?

A

amino acid tryptophan

103
Q

What bacterial enzyme produces indole?

A
  • tryptophanase
104
Q

Phenol red vs Methyl Red

A
  • PR= orange-neutral; yellow-acidic
  • MR= yellow-neutral; red-acidic
105
Q

Why do some cells that lack flagella appear to move?

A
  • Brownian motion
  • due to water molecules continuously striking them
106
Q

Why Levin EMB is selective medium and Luisetti’s is not?

A
  • EMB contains dyes that inhibit the growth of Gram +
  • Luisettis is not b/c many species grow in medium
107
Q

Why Levin EMB is considered differential medium?

A
  • Lactose-Gram Negative
  • Some will ferment some will not
  • e.coli & e.aerogenes.
108
Q

Why organisms can survive/org under aerobic conditions need enzymes and superoxide dismutase

A
  • to protect itself from ROS
109
Q

Fungi

A
  • Mycology
  • Eukaryotes
  • Myceteae kingdom
  • hetertrophs, saprobes
  • decomposers
  • reproduction-spores
110
Q

Unicellular Fungi

A
  • yeast and mildew
111
Q

Macroscopic Colonial fungi

A
  • fuzzy
  • form molds
  • long filamentous cell bodes, hyphae
112
Q

Multi cellular fungi

A
  • form mushrooms.
113
Q

chitin

A
  • thick and rigid walls of mushrooms
  • polysaccharide of N-ace
114
Q

Saprobes

A
  • obtain nutrients from dead plants/animals
115
Q

Sabouraud Dextrose Agar

A
  • selective medium for fungi
  • contains acid and inhibits growth of glucose
116
Q

Sporangia

A
  • spore producing structures
117
Q

Gas Test

A
  • -A/-G= Red
  • +A/-G= Red to Yellow, no bubbles
  • +A/+G= Red to Yellow, bubbles

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