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BIO233 > Observing Microbes & Microbial Structure > Flashcards

Observing Microbes & Microbial Structure Flashcards

1
Q

Nano meter compared to micrometer

A

1 micro meter = 1/1000 nanometer

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2
Q

1 micrometer = X meters

A

10 to the power of -6

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3
Q

1 nanometer = X meters

A

10 to the power of -9

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4
Q

E. coli size comparison to RBCs

A

RBCs are bigger

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5
Q

Microscope capable of seeing the smallest thing

A

Transmission electron microscope

10 picometers

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6
Q

Pathway of light through compound light microscope

A

Illuminator
condensor
Specimen
Objective lens
Ocular lens

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7
Q

Total magification of compound microscope=

A

Magnification of objective lens (4-100x) x magnification of ocular lens (10x)

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8
Q

Microscope objective

A

the different magnification strengths on a microscope

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9
Q

Aperture seettings on a light microscope affect

A

contrast and resolution

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10
Q

Resolution

A

The ability to distinguish bw two points or to see fine details

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11
Q

The higher the magnification of the objective the more _____ is required

A

Light

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12
Q

Longer or shorter waveslengths of light producer greater resolution

A

shorter

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13
Q

Highest resolution of light microscopes

A

.2 micrometers (2000x magnification)

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14
Q

What can be seen using a light microscope

A

Bacteria and fungi

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15
Q

Two forms of light microscopy

A

Bright field
Dark field

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16
Q

Bright field microscopy

A

Direct light enter objective lens
Dark objects visible against bright/white background
Light reflected off the specimen
Staining increases contrast

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17
Q

Is staining a part of bright field or darkfield

A

Bright field

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18
Q

Darkfield microscopy

A

Opaque disk placed bw light and specimen
- Only light reflected (diffracted) by specimen reaches eyepiece
- Emphasises edges of structures against a dark background
- The background is stained, not the object itself

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19
Q

Fluorescent microscopy

A

Uses UV light or other short wavelengths of light

Substances absorb UV light and emit visible light (longer wavelengths)

M/os stained with antibodies combined with fluorescent dyes (fluorochromes

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20
Q

How is bacteria identified under a microscope via fluorescent microscopy

A

To identify unknown bacteria, antibodies specific to the bacteria and fluorochrome are combined and attach to bacteria to light it up

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21
Q

What is lysteria

A

Organisms that contaminates foods in the fridge section, invades host cells and uses the host cell to propel them to the next cell

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22
Q

Two types of elctron microscopy

A

TEM: Transmission electron microscopy
SEM: Scanning electron microscopy

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23
Q

What type of images produced by electron microscopy

A

Black and white

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24
Q

TEM

A

Ultrathin slice of specimen

2D image of internal structures (The best microscope to see internal structures)

Treatment can cause distortion (Thin slicing)

Magnification of 10,000 - 100,000x
Resolution 2.5 nm

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25
Q

Path of light through TEM

A

Electron beam through electromagnetic lens - specimen - electromagnetic lens - screen or film

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26
Q

Why do beam of electrons produce higher resolution than light or UV light

A

shorter wavelength than visible and UV light; therefore, greater resolution

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27
Q

How are specimens stained in electron microscopy

A

Metal salts (eg. Lead & Uranium salts)
Immuno-staining (antibodies coated with gold particles)

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28
Q

SEM

A

Electrons produced by electron gun

scans whole surface of specimen (3D image)
- Useful for studying surface structures

Electrons being scattered off specimen are captured by microscope

Less distortion bc no slicing

1000-10000x magnification
Resolution of 20nm

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29
Q

Does SEM or TEM have higher resolution

A

TEM is 2.5 while SEM is 20nm

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30
Q

What size of specimen is a microscope needed for

A

1mm or smaller

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31
Q

Stains are used to increase contrast in what kind of microscopy

A

Electron

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32
Q

Most frequently used differential stain

A

Gram stain

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33
Q

Characteristics of simple stains

A

They are basic and can be rinsed with water

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34
Q

Most stains used in microbiology are _______ tyoe of stains

A

differentialho

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35
Q

how many steps to gram staining

A

7

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36
Q

Best microscope for visualizing flagella

A

Electron microscope

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37
Q

Why is staining important

A

Most Organisms can be colourless, and therefore will be invisible under the microscope

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38
Q

Why are salts used for staining

A

Bacteria cells are negative and attract the positive ions of salts

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39
Q

How do stains work

A

One of the ions is colored (chromophore)

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40
Q

WHy are bacteria slightly negative

A

Bc of they are phospholipids

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41
Q

Chromophore

A

The stained ion in the salt

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42
Q

Basic dyes (simple stains)

A

Most common dyes
Chromophore is cation (therefore binding to negative bacteria)

Includes crystal violet, methylen blue, safranin

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43
Q

Acidicdyes

A

Chromophore is an anion
Used for negative staining (bacteria remains colourless while background is stained)

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44
Q

3 types of stains

A

simple
differential
special

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45
Q

Simple stains

A

Stains the cell
Increases contrast

Includes crystal violet, methylen blue, safranin

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46
Q

Differntial stains

A

Using more than one type of dye

Differentiate bw organisms or structures

Gram stain MOST FREQUENTLY USED (Differentiates gram neg bacteria from gram positive)

acid fast stains (bw acid fast and non acid fast)

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47
Q

Special stains

A

(isolates specific groups)
Used to differentiete structures

Capsule (negative since capsules remain unstained, instead halo appears around them), endospore (stain to detect presence of), flagella stains

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48
Q

Gram staining

A

A type of differential staining

  • Gram positive bacteria: stainined purple (Crystal violet/iodine)
  • Gram negative bacteria: stained red by safranin (counterstain)
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49
Q

Steps of gram staining

A

Application of crystal violet (wait 60 seconds)
Rinse
Add iodine (mordant = reactive to crystal violet to cause greater adhesion to the stain)
Rinse off excess mordant
Alcohol wash (decolorization)
To wash off any dye not strongly held to cells
Application of safranin (counterstain)

Those that lost the crystal violet will be stained red; however, those already purple will be unaltered by the additional pinkish/red dye

Purple cells = gram positive
Pink/red cells = gram negative

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50
Q

Gram positive cells

A

More peptidoglycan
When dye added, the crystal violet attaches throughout the layer peptidoglycan layer TO THE PLASMA MEMBRANE WHICH IS NEGATIVE

When decolorizer is added, peptidoglycan is compressed and locks in the dye in

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51
Q

Gram negative cells

A

Cell wall Made up of two membranes
In bw two membranes is thin single peptidoglycan layer

Crystal violet attaches to outer membrane and some trickles into peptidoglycan
Decolourizer results in gram violet dye escaping outer lipid layer

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52
Q

What colour is a gram positive stain

A

Purple of blue

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53
Q

What colour is the gram negative stained cells

A

Red/pink

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54
Q

Important gram positive bacteria

A

Corynebacterium diphtheriae

Staphylococcus aureus

Streptococcus pyogenes

Streptococcus pneumoniae

Gardnerella vaginalis (gram variable)

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55
Q

Important gram negative bacteria

A

Escherichia coli

Neissseria gonorhoeae

Gardnerella vaginalis (Gram varariable)

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56
Q

Stains used in acid fast staing

A

Red and blue

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57
Q

Acid fast bacteria appear what colour under staining

A

Red

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58
Q

Non acid fast bacteria appear what colour under staining

A

BLue

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59
Q

What identifies a bacteria as acid fast

A

Acid fast bacteria have thick outer lipid layer

Mycolic acid

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60
Q

Examples of important acid fast bacteria

A

Mycobacterium tuberculosis (TB)
Mycobacterium leprae (leprosy)
Process

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61
Q

Process of acid fast staining

A
  • Primary stain is carbolfuschin (red dye) - CARBOLFUSCHIN stains the mycolic layer
  • Decolorized - by acid alcohol
  • Anything without mycolic layer dye is washed off
  • Counter stain methylene blue is used (Stains the plasma layer)
  • Non-acid fast stained blue
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62
Q

What is the primary stain in acid fast staing

A

Carbolfuchsin (red dye),

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63
Q

Who proposed three domain system

A

Carl Woese

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64
Q

three domains

A

Bacteria, Archaea & Eukarya
Three distinctly different cell groups based on ribosomal RNA (rRNA)

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65
Q

Basic defintion of bacteria

A

Pathogenic and non pathogenic prokaryotes

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66
Q

Basic def of archaea

A

Prokaryotes live in extreme environments, carry out unusual metabolic processes
Extreme halophile and hyperhermophiles (habitatish)

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67
Q

Basic def of Eukarya

A

animals, plants, fungi;

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68
Q

Characteristics of bacteria

A

Prokaryotic
Contains peptidoglycan
Straight carbon chains
Sensitive to antibiotics

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69
Q

Characteristics of Archaea

A

Prokaryotic
Does not contain peptidoglycan
Varies in composition
Not sensitive to antibiotics

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70
Q

Characteristics of Eukarya

A

Vary in composition
Not reactive to antibiotics

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71
Q

Prokaryotes vs Eukaryotes

A

Prokaryotes
One circular chromosome, not membrane bound
No histones
No membrane bound organelles
Peptiodclycan cell walls (bacteria)
Divide by binary fission

Eukaryotes
Paired chromosomes
Histones (DNA wrapped around proteins called histones)
Membrane bounded organelles
Polysaccharide cell wall
Division via mitosis
LARGER than prokaryotes

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72
Q

Structures found in all bacteria

A

Plasma membrane
Cytoplasm
Ribosomes
Nucleoid w/ DNA

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73
Q

Can bacteria be infected by viruses

A

Yes

74
Q

Basic shapes of bacteria

A

Coccus, bacillus, spirillum (rigid), spirochetes (flexible) ALSO intermittent (something in bw two structures)
Shapes determined by cell wall structure

75
Q

Flagellin

A

Forming helical chains that make up the filament in the flagellum

76
Q

Difference bw flagellum in eukaryotes vs prokaryotes

A

Eukaryotic flagella is covered by a membrane

77
Q

Pathogenic bacteria can be identified by

A

differences in flagerllar proteins

78
Q

Basal body similar to what in a car

A

Driveshaft

79
Q

Cell division in Cocci bacteria

A

Can divide on many planes
Diplococci
Streptococci (Chain of spherical cells)
Staphylococci (CLuster of spherical cells)

80
Q

Cell division in bacillus

A

Cell division can involve just one plane, hence fewer arrangements:
Pairs: Diplobacilli
Chains: Streptobacilli
Coccobacillus:

81
Q

Vibrio

A

Curved shapes

82
Q

Spirullum

A

Helical shape (corkscrew)
- Rigid structure

83
Q

Spirochetes

A

Helical shape but flexible
Propelled by an internal axial filament

84
Q

Other shapes bacteria can take

A

Star or rectangualr

85
Q

Monomorphic

A

Maintaining a single state (most bacteria are this)

86
Q

Pleoorphic

A

Having many shapes, usually if lacking a cell wall

87
Q

Two meanings of bacillus

A

can be genus AS WELL AS shape

88
Q

Do all bacteria have a cell wall

A

A few of them are pleomorphic and thus DO NOT

89
Q

Main external structures of prokaryotes

A

Pilia and fimbria
Glycocalyx
Flagella

90
Q

GLycocalyx

A

(outside structure mostly contained of carbs)

External to the cell wall
Extracellular polysaccharide

Usually sticky

Capsule: neatly organized and tightly packed, preventing phaogcytosis. MAKES THE bacteria INVISIBLE to phagocytes

Slime layer: organized and loose

91
Q

Two parts of glycocalyx

A

Capsule and slime layer

92
Q

Flagella

A

Made of chains of flagellin
Attached to a protein hook
Anchored to the wall and membrane by the basal body

93
Q

What can alter the speed and direction of rotation of flagella

A

Bacteria

94
Q

4 kinds of flagella arrangements

A

Peritichous (Attatchment around the perimeter)
Monotrichous
Lophotrichous(several bunched arising from one end) and polar
Amphitrophous (Flagella at both ends)

95
Q

What is a run for a a bacteria with a flagella

A

Movement of bacteria by flagellar propulsion (counterclockwise)

also called a random walk

96
Q

Flagella tumbling

A

Flagella tumbling: When one flagellum spins in the other direction (Clockwise) causing the whole rhythm to be mixed up and random tumbling

97
Q

Biased random walk

A

Resulting from a chemotxis agent, the movement of a cell toward an attractive agent that results in less tumbling than a random walk

98
Q

Chemotaxis

A

Using chemicals as a way to direct cells somewhere
Random walk determined by rotation of the flagella
Run is moving in one direction (counterclockwise)
Tumbling is spinning into a new direction (Caused by clockwise)

99
Q

Spirochete

A

Spirochete bacteria have their flagella embedded in hte middle of the membrane

100
Q

Fimbriae and pili

A

Fimbraie allow attachment
Pili (Pilus) are used to transfer DNA from one ell to another (Longer than fibraie)

Fimbriae is one of the way diseases are spread
UTIs can begin by fimbriae of bacterial cells attaching in geneital area

101
Q

Which part of the phospholipid bilayer is hydrophobic

A

The fatty acid tails

102
Q

Axial filaments are composed of

A

Endoflagella

103
Q

Streptobacillus

A

Chains of rods

104
Q

Correct order for gram staining

A

Primary stain
Mordent
Decolorize
Counterstain

105
Q

What is the difference bw the condensor and the diaphragm on the microscope

A

Diaphragm controls amount of light entering condensor

Condensor focuses light thorugh specimen

106
Q

Objects smaller than _____ must be observed through an electron microscope

A

0.2 micrometers

107
Q

Disadvantages of TEM

A

No 3D aspect
specimens must be fixed, dehydrated, and viewed under a high vacuum to prevent electron scattering. These treatments not only kill the specimen, but also cause shrinkage and distort

108
Q

Smear

A

a thin film of material containing the microorganisms is spread over the surface of the slide

109
Q

Benefits of negative staining

A

valuable for observing overall cell shapes, sizes, and capsules because the cells are made highly visible against a contrasting dark background

Dostrotions mimized bc no fixing necessaru

110
Q

Mordant

A

a chemical is added to the solution to intensify the stain

111
Q

Malachite green stain is used

A

To detect the presence of endospores in bacteria

112
Q

Is crystal violet a basic or acidic dye>

A

Basic

113
Q

Microscope that produces colour images without staining using two beams of light

A

DIC

114
Q

Affect of Gram’s iodine on gram positive cells

A

Forms complexes with crystal violet aking it more difficult for it to wash out

115
Q

How does the age of a culture affect gram staining?

A

. A Gram-positive culture more than 24 hours old contains a substantial fraction of old cells with a thin peptidoglycan layer and may Gram stain pink.

116
Q

How much of the culture should be sampled?

A

The loop should barely swipe the surface of an isolated colony to ensure that the smear is made to the correct level of thickness.

117
Q

What will happen if broth is not resuspended prior to sampling>

A

most cells will settle at the bottom of the tube, leaving few cells at the top of the tube.

118
Q

How is preparing a solid culture different from a liquid one

A

During smear preparation from solid cultures, a droplet of water must first be added to the slide and then mixed with the cells.

119
Q

What is used to view internal cell structures in their native state

A

phase-contrast microscope

120
Q

Which of the more high powered microscopes uses visible light?

A

differential interference contrast microscope

121
Q

Which microscope allows visualization of atoms

A

scanning tunneling microscope

122
Q

How can intracellular detail be viewed in a LIVING cell

A

Two-photon microscope

123
Q

The order of making a smear

A

Make it
Fix it
Stain it

124
Q

What is fluoresence microscopy

A

microorganisms stained with a fluorochrome are examined under a fluorescence microscope with an ultraviolet or near-ultraviolet light source, they appear as luminescent, bright objects against a dark background.

125
Q

How does fluorecent microscopy work

A

Antibodies for a specific bacteria combine with fluorochrome and then bind to the bacteria, lighting it up

126
Q

Scanning Acoustic Microscopy

A

basically consists of interpreting the action of a sound wave sent through a specimen

127
Q

What does fixing a specimen imply

A

by passing it through the flame of a Bunsen burner several times, smear side up, or by covering the slide with methanol for 1 minut

128
Q

Forgetting to fix a specimin

A

Will result in it washing away

129
Q

Gram positive bacteria react to dye how

A

By retaining purple colour

130
Q

Are gram positive or negative bacteria more resitient to antibiotics

A

Gram negative

131
Q

What is alcohol used for in the gram staining process

A

Decolorization

132
Q

Which type of stain is used to identify the bacterial agents responsible for tuberculosis and leprosy?

A

Acid fast staining

133
Q

What is an endospore

A

special resistant, dormant structure formed within a cell that protects a bacterium from adverse environmental conditions

134
Q

Longer, outmost region of flagella

A

Filament

135
Q

Flagellin

A

Globular protein arranged in several chains that intertwine and form a helix around a hollow core.

136
Q

Hook of the flagella

A

Slightly wider part of flagella attatching it to basal body

137
Q

Basal body of flagella

A

anchors the flagellum to the cell wall and plasma membrane

138
Q

How is fimbrae relevant to pathogens?

A

Fimbriae is one of the way diseases are spread
UTIs can begin by fimbriae of bacterial cells attaching in geneital area

A bacteria is determined to be virulent by the fimbriae

139
Q

Example of bacteria with large capsule

A

Streptococcus pneumonia has a large capsule

140
Q

purpose of capsule

A

The capsule makes the bacteria invisible
Can prevent antibiotics from entering bacteria
Protects bacteria

141
Q

Purpose of cell wall

A

Gives bacteria its shape
Prevents osmosis lysis

142
Q

Osmosis

A

Water moving from high to low conc across cell membrane

143
Q

Is water conc higher inside or outside the cell

A

Outside, bc of dissolved solutes inside the cell; therefore water flows in

144
Q

Cell wall in gram + bacteria

A

Composed of peptidoglycan

Repeating NAG-NAM units (long chain of polysachharides)
Crosslinked by peptides, teichoic acid (holds the wall together), and lipteichoic acid (anchors the wall to the plasma membrane)

145
Q

Cell wall in gram - bacteria

A

Cell wall made of pepdidoglycan and outer membrane in G-

Cell wall (Outer membrane + peptidoglycan layer) = protection from phagocytes, complement, and antibiotics

Porins (made of proteins) form channels from inter membrane to outer to allow for easier crossing of the cell wall

146
Q

Penicillin most affective against which type of gram bacteria

A

Gram positive bc penecillin affects peptidoglycan

Blocks peptidoglycan cross-linking of cell wall

147
Q

Lysozymes effect on bacteria cells

A

Breaks peptidoglycan
Sleep in the eyes is often dead bacteria cells

148
Q

Penecillin affect on gram - bacteria

A

Pepditodglycan layer sndwhiched, so penicillin cannot easily get there to affect it
Penecillin is hydrophilic, and the inner layers of membrane are hydrophobic

149
Q

Important Gram + bacteria

A

Corynebacterium diphtheriae
Staphylococcus aureus
Streptococcus pyogenes
Streptococcus pneumoniae
Gardnerella vaginalis

150
Q

Important Gram - Bacteria

A

Escherichia coli
Neissseria gonorhoeae
Gardnerella vaginalis

151
Q

Saccharide

A

Sugar

152
Q

What differentiates Gram - bacteria

A

Lipopolysaccharide within gram negative walls

153
Q

Three parts of Lipopolysaccharide

A

lipid A buried in the outer membrane (is an endotoxin)
Core polysaccaride
Connects to the O polusaccharide tail

154
Q

Endotoxin vs exotoxin

A

Endotoxin is only released to become a toxin when the cell breaks down (destroyed)
Exotoxins are only produced by Gram + that are released to the outside without the bacterias death

155
Q

Which type of gram bacteria are antibiotics potentially dangerous for treating?

A

ANtibiotics can be dangerous to give to those with Gram - bacteria infections bc toxins can be released

156
Q

Mycoplasmas

A

Type of microorganism with an atypical “cell wall”

Lack cell walls (THEREFORE UNAFFECTED BY ANTIBIOTICS)
Pleomorphic
Sterols in plasma membrane (Cholestrol etc. help keep membrane structure intact)

157
Q

Acid fast cell walls

A

Like Gram-positive
Waxy lipid (mycolic acid)
Cells grow slowly bc of wax layer
Mycobacterium, nocardia

158
Q

Parts of phospholipid bilayer

A

Outer is hydrophilic heads
Inner is hydrophobic (tails)
Excludes many things that are hydrophilic
Associated proteins

159
Q

Associated proteins with phospholipid bilayer

A

Perpheral proteins: Not deep/tightly bound, can be stripped from the membrane

Integral proteins: Imbeddded in the membrane, only remobved through destroyign the membrane structure

Transmembrane proteins: Cross the membrane (porins), allow for stuff to cross the membrane (transport proteins, channels etc.)

160
Q

Carbohydrates in microorganisms

A

Located on the outside of the membrane, attached to proteins OR lipids
Used for energy
Cell communication (When cell communication stops it results in cancer cells)
Cell identification

161
Q

What are granules in the cytoplasm for?

A

Primarily phosphate and energy reserves

162
Q

Endospores

A

Highly durable dehydrated dormant cells (withstands boiling for up to 20 hours)
Allows cells to survive adverse conditions

163
Q

Formation of endospores called

A

Sporation

164
Q

Endospore germination

A

AN ENDOSPORE returning to a vegetative (growing) cell
One endospore gives rise to one bacteria cell

165
Q

Medical significance of endospores

A

Baciculls antrhacis - the cause of anthrax
Clostridium (gram positive therefore it is an exotoxin)

166
Q

Major kinds of endospore cholstridium

A

Tetanus (contracts muscles)
Botulism (relaxes muscles) - used for BOTOX
Gas gangrene
C. difficile

167
Q

What protein composes flagella in eukaryotes vs prokaryotes

A

Tubulin in eukaryotes
Flagellin in prokaryotes

168
Q

How can pathogens be identified by their flagellum?

A

Differences in their flagellar proteins

169
Q

Glycoproteins

A

Proteins attatched to carbohydrates

170
Q

Pili are used for

A

Transfer of DNA and motiility

171
Q

Best description of how streptomyocin attacks bacterial cells

A

Streptomycin targets the 70S ribosome and inhibits protein synthesis

172
Q

Why are most Gram-negative bacteria resistant to the actions of penicillin?

A

Penicillin is unable to pass through the outer membrane of the cell wall.

173
Q

What part of E. coli are responsible for the symptoms of a UTI

A

Fimraie

174
Q

A gram-positive bacterium suddenly acquires resistance to the antibiotic methicillin. This trait most likely occurred due to acquisition of new genetic information through

A

conjugation

175
Q

How do spirochetes and spirilla differ?

A

Spirilla have external flagella, but spirochetes have axial filaments.

176
Q

Which kind of gram cells have porins?

A

Gram - bacteria

177
Q

Explain Acid fast staining

A
178
Q

What does the Gram Positive cell wall NOT protect the bacteria from

A

Does not prevent cell lysis in hypertonic environment

179
Q

A bacterium that an grow in the presence or absence of oxygen produces energy by

A

Glycolysis, Krebs cycle, and oxitative phosyphorilation

180
Q
A

BIO233 (18 decks)

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