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Pre-health Microbiology > Lab Midterm Lecture Info > Flashcards

Lab Midterm Lecture Info Flashcards

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Biology 101 flashcards
1
Q

first person to observe microbes, including bacteria, which he called “animalcules”

A

Antonie van Leeuwenhoek, 1600s

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2
Q

the use of any kind of microscope that uses visible light to observe specimens

A

light microscopy/compound microscopy

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3
Q

see living organisms, motility, bright objects on a dark background

A

dark-field microscopy

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4
Q

blocks most of the light from the illuminator in dark field microscopy

A

opaque disk

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5
Q

the only light that reaches the objective in dark field microscopy

A

refracted or reflected light by structures in the specimen

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6
Q

causes syphilis

A

treponema pallidum

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7
Q

two sets of light- one directly from the light source, one from light that is reflected or diffracted from a structure in the specimen

A

phase contrast microscopy

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8
Q

structures that differ in features such as ___ ___ will differ in levels of darkness; phase microscopy

A

refractive index

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9
Q

example- 40x objective lens
a. E- type apochromatic lens
b. 40x magnification
c. numerical aperture of 0.65
d. 160 mm mechanical tube length
e. 0.17 mm thickness cover slip

A

important markings on a light microscope objective lense

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10
Q

___ lenses are made in such a way that chromatic aberration is reduced to a minimum

A

apochromatic

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11
Q

describes the capacity of a microscope to enlarge an image
-objective and ocular

A

magnification

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12
Q

the ability to distinguish two adjacent objects as distinct and separate

A

resolution

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13
Q

light-gathering ability of the objective lense

A

numerical aperture

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14
Q

__ wavelength = better resolution

A

shorter wavelength

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15
Q

limit of resolution for a light microscope is about __ um

A

0.2

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16
Q

objects closer together than this value cannot be resolved as distinct and separate

A

limit of resolution

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17
Q

magnification of ocular lense

A

10x

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18
Q

D = wavelength / NAcondenser + NAobjective

A

formula for calculating the actual limit of resolution for a microscope

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19
Q

has the same refractive index as glass

A

immersion oil refractive index

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20
Q

increases the maximum angle at which light leaving the specimen can strike the lense

A

immersion oil

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21
Q

mostly UV or blue light

A

light source of fluorescent microscopy

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22
Q

uses an electron beam to create an image, with electromagnets acting as lenses

A

electron microscopy

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23
Q

image generated using flurescence

A

fluorescent microscopy

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24
Q

uses electron beams to observe small, thin specimens such as tissue sections and sub-cellular structures

A

transmission electron microscope (TEM)

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25
Q

uses electron beams to visualize surface 3D surface details of specimens

A

scanning electron microscope (SEM)

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26
Q

used to clean all lenses

A

dry, clean lens paper

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27
Q

used to remove oil from the stage

A

ethanol

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28
Q

-low-power objective in position and body tube lowered completely
-centered mechanical stage

A

proper set up of microscope

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29
Q

coil the electric cord around the body tube and the stage

A

proper cord position during microscope transfer

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30
Q

three types of bacterial morphology

A

cocci, bacilli, spiral

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31
Q

example of diplococci

A

streptococcus pneumonia, Enterococcus

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32
Q

example of cocci clusters

A

staphylococcus aureus

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33
Q

example of cocci chains

A

streptococcus pyogenes

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34
Q

example of flagellate rods

A

salmonella typhi

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35
Q

example of bacilli chains

A

bacillus anthracis

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36
Q

example of spore formers

A

clostridium botulinium

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37
Q

example of spirochetes

A

treponema pallidum morphology

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38
Q

example of spirilla

A

helicobacter pylori

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39
Q

example of vibrios- aka curved rods

A

vibrio cholera

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40
Q

will occur if a culture plate is left open on a lab bench

A

microorganisms will contaminate

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41
Q

practices and procedures to prevent contamination from pathogens and minimize the risk of infection

A

aseptic technique

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42
Q

6 inches

A

stay within ___ inches of bunsen burner to minimize contamination

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43
Q

a liquid medium

A

broth

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44
Q

usually made with 1.0 % - 2.0% agar in plates or tubes

A

solid medium

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45
Q

usually made with 0.3%-0.5% agar in plates or tubes

A

semi-solid medium

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46
Q

refers to cultivating and growing microorganisms in the lab on various types of media

A

culturing

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47
Q

-solidifying agent
-allow surface growth or restrict mobility
-grow bacteria over a range of temperatures
-liquifies at 100 C and solidifies at ~42C

A

agar

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48
Q

organism, name, section number, date

A

tube labels

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49
Q

how to sterilize an inoculating loop/needle

A

-hold in blue cone of flame at 45 degree angle
-let cool before transfer
-re-flame when finished and place upright back in block container

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50
Q

considered contaminated at all times

A

wire holder of inoculating loop/needle

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51
Q

color of inoculating needle/loop for sterilizing in flame

A

orange/red incandescence

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52
Q

-abundance of growth
-pigmentation
-optical characteristics
-form

A

microbe culture characteristics of agar slants

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53
Q

-growth type and distance from stab
-pigmentation or appearance

A

microbe culture characteristics seen in agar deep tubes

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54
Q

-abundance of growth
-type of growth

A

microbe culture characteristics seen in broth medium

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55
Q

transfer from agar slant to sterile broth tube

A

use loop

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56
Q

from broth stock to sterile agar deep tube

A

use needle and stab inoculation

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57
Q

from broth stick to sterile agar slant tube

A

use loop and inoculate surface of slant

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58
Q

incubation temperature of bacillus cereus and Serratia marcescens

A

25 degrees C for 24 to 48 hours, exercise 2

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59
Q

incubation temperature for staphylococcus epidermidis and Pseudomonas aeruginosa

A

37 degrees C for 24 to 48 hours, exercise 2

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60
Q

bacteria (pure or mixed culture) are diluted until single cells are separated from one another/isolation of distinct colonies

A

streak plate definition (4 quadrant streak)

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61
Q

-to grow into isolated pure colonies/determine purity of culture
-color, morphology and other physical characteristics
-quick first step in the identification of the bacteria being studied

A

purposes of streak plate

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62
Q

heavy confluent growth, light growth, discrete colonies

A

types of growth on streak plate

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63
Q

streak plate but with 3 sections instead of 4

A

t-streak

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64
Q
  1. sterilize loop
  2. touch loop to bacterial culture (broth or colony)
  3. streak heavy in one quadrant
  4. flame loop
  5. streak in and out of 1st quadrant a few times into 2nd quadrant then only streak in 2nd quadrant to fill
  6. repeat steps for the 3rd and 4th quadrants
  7. when done, sterilize loop
A

procedural steps for streak plate- isolating discrete bacterial colonies

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65
Q

-form
-elevation
-pigmentation/color
-size
-optical properties

A

microbe culture characteristics of agar plates

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66
Q

colonies are mucoid, raised, and shiny

A

Klebsiella pneumoniae agar plate properties

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67
Q

sample is pipetted onto surface of agar plate, sample is spread evenly over surface using sterile glass spreader, surface colonies seen

A

spread-plate method

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68
Q

sample is pipetted into sterile plate, sterile medium is added and mixed well, surface and sub-surface colonies seen

A

pour-plate method

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69
Q

24 to 48 hour nutrient broth cultures of a mixture of E. coli and B. subtilis

A

cultures for exercise 3

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70
Q

two trypticase soy (TS) plates per student

A

media used for exercise 3

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71
Q

incubation temperature for exercise

A

37 degrees celcius

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72
Q

practice aseptic technique to transfer cultures
-agar slant to terile broth tube (loop)
-broth to agar deep tube (needle and stab)
-broth to agar slant (loop surface)

A

exercise 2 culture transfer

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73
Q

perform two 3- or 4-quadrant streak plates and two spread plates using a mixed organism broth culture

A

exercise 3- isolating distinct colonies

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74
Q

picking up a single isolated colony

A

pure culture

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75
Q

to prepare a stock culture of an organism using isolates from the mixed cultures prepared on the agar streak plate and or the spread plate in exercise 3

A

purpose of exercise 4- preparation of pure cultures

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76
Q

solution consisting of a solvent (usually water or ethanol) and a colored molecule (often a benzene derivative)- the chromogen

A

stains

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77
Q

positive chromogen, stains cell

A

basic stain

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78
Q

negative chromogen, background is stained

A

acidic stain

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79
Q

auxochrome

A

provides covalent or ionic bonds in stain

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80
Q

colored compound, benzene (colorless) and chromophore (imparts color)

A

chromogen

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81
Q

developed the gram stain

A

Hans Christian Gram 1884

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82
Q

appear purple after staining

A

gram-positive bacteria

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83
Q

appear pink after staining

A

gram negative bacteria

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84
Q

differences in cell wall structure

A

reason for color difference in gram stain

85
Q

has thick cell wall

A

gram-positive bacteria structure

86
Q

has thin cell wall, with outer membrane

A

gram-negative bacteria structure

87
Q
  1. primary stain- crystal violet
  2. mordant- iodine
  3. decolorizing agent- alcohol-acetone
  4. counterstain- safranin
A

order of application for gram stain

88
Q

appears colorless after decolorizing agent

A

gram-negative cells

89
Q

contains notable human pathogens such as M. tuberculosis, M. leprae

A

genus Mycobacterium

90
Q

gram-positive bacteria that are acid-fast because of the waxy mycolic acid in their cell walls

A

mycobacteria

91
Q

detects the presence of cell walls rich in mycolic acid

A

acid-fast staining protocol (Ziehl-Neelsen Method)

92
Q

stains everything strongly in acid-fast staining

A

carbolfuchsin

93
Q

decolorizing agent in acid-fast staining

A

acid alcohol

94
Q

counterstain in acid-fast staining
-stains non acid-fast cells blue

A

methylene blue

95
Q

structures that protect the bacterial genome in a dormant state when environmental conditions are unfavorable

A

endospore

96
Q

bacillus and clostridium

A

endospore-forming, gram-positive bacteria genera

97
Q

the process by which vegetative cells transform into endospores

A

sporulation

98
Q

staining detects endospores

A

Schaeffer-Fulton method

99
Q

malachite green and carbolfuchsin

A

need steam to enter cells for staining

100
Q
  1. primary stain- malachite green
  2. spore retains malachite green while bacterias structures loose the stain- spores resist decolorization with water
  3. counterstain- safranin
A

process of spore staining- Schaeffer-Fulton Method

101
Q

non-specific media configured to culture a wide range of microorganisms without many restrictions

A

general growth media

102
Q

designed to suppress the growth of unwanted bacteria and encourage the growth of the desired microbes

A

selective media

103
Q

make it easier to distinguish colonies of the desired organism from other colonies growing on the same plate

A

differential media

104
Q

Nutrient agar and Luria-Bertani (LB) Broth

A

examples of general growth media

105
Q

similar to selective media but designed to increase numbers of desired microbes to detectable levels

A

enrichment

106
Q

both selective and differential for staphylococcus aureus

A

mannitol salt agar

107
Q

high salt concentration (7.5% NaCl) inhibits growth of most bacteria except staphylococci

A

what makes MSA selective for staphylococci

108
Q

carbohydrate mannitol is fermented by S. aureus, resulting in acidic end products- turns phenol red indicator yellow around growth

A

MSA differential reaction of staphylococcus aureus

109
Q

microbiologists often use this to identify bacterial species that destroy red blood cells

A

blood agar

110
Q

alpha (a) hemolysis

A

partial/incomplete lysis of RBC, zone of partial clearing = green halo around colonies

111
Q

beta (B) hemolysis

A

complete lysis of RBC, complete zone of clearing around colonies

112
Q

gamma (Y) hemolysis

A

no lysis of RBCs, no clearing of medium surrounding colonies, no color change

113
Q

tends to be used to recover fastidious bacteria, often Streptococcus species (pathogens)

A

blood agar bacterial species

114
Q

blood agar is a ___ media, bacteria distinguished by ability to cause hemolysis

A

differential media (hemolysis)

115
Q

selective and differential medium containing lactose, bile salts, neutral red, and crystal violet

A

MacConkey Agar

116
Q

interferes with the growth of many gram-positive bacteria and FAVORS gram-negative bacterial growth, especially Enterobacteriaceae

A

MacConkey agar favorable growth

117
Q

named enterics, reside in the intestine, are adapted to the presence of bile salts

A

Enterobacteriaceae

118
Q

ph indicator in MacConkey agar, colorless above a pH of 6.8 and red at a pH below 6.8
- acid accumulating from lactose fermentation turns dye red

A

neutral red dye

119
Q

-used for the isolation and identification fo fecal coliform bacteria
-sugars act as fermentable substrates, which yield acid by-products
-dyes inhibit growth of gram-positive bacteria and act as pH indicators (only used to see gram-negative)

A

Eosin Methylene Blue Agar

120
Q

eosin and methylene blue inhibit growth of gram-positive bacteria

A

why eosin methylene blue agar is selective

121
Q

distinguishes gram-negative bacteria that can ferment lactose from those that cannot

A

why eosin methylene blue agar is differential

122
Q

lactose fermenters in eosin methylene blue agar

A

produce dark colonies- metallic green shade

123
Q

non lactose fermenters in eosin methylene blue agar

A

produce opaque or translucent colonies

124
Q

example of lactose fermenter

A

Escherichia. coli

125
Q

small amounts of acid production in eosin methylene blue agar

A

results in pink coloration of the growth

126
Q

an undefined, selective medium that allows growth of gram-positive organisms and stops or inhibits growth of most gram-negative orgamisms

A

phenylethyl alcohol agar

127
Q

false negative results and low colony counts

A

viable but non-culturable (VBMN)

128
Q

bacterial species that will grow within a temperature range of -5°C to 20°C
- all will grow between 0° and 5°

A

psychrophiles

129
Q

bacteria species that will grow within a temperature range of 20°C to 45°C
-all can grow at human body temperatures (37°C) and are unable to grow above 45°C
-three distinct groups

A

mesophiles

130
Q

optimal growth at 20-30°C
-mesophile

A

plant sacrophytes

131
Q

optimum growth temperature at 35-40°C
-mesophile

A

organisms that grow in warm blooded hosts

132
Q

optimum growth at 20-40°C but are capable of growing at 0°, typically found in soil and water habitats in temperate regions
-mesophile

A

psychrotolerant

133
Q

bacterial species that will grow at 35°C and above
-two groups exist

A

thermophiles

134
Q

thermophiles that will grow at 37°C but grow optimally at 45-60°C

A

facultative thermophiles

135
Q

thermophiles that will grow only at temperatures above 50°C, optimum growth above 60°C

A

obligate thermophiles

136
Q

detection of gas accumulation

A

air bubble in durham tube (within a culture tube)

137
Q

enzyme that protects cell from toxic H2O2- hydrogen peroxide

A

catalase and peroxidase

138
Q

enzyme that protects cell from toxic O2^- superoxide

A

superoxidedismutase

139
Q

has no enzymes to protect against toxic oxygen- cannot grow in its presence

A

strict anaerobes

140
Q

-requires the presence of atmospheric oxygen for growth
-their enzymatic needs to use oxygen as the final electron acceptor

A

aerobes

141
Q

microaerophiles

A

-require limited amounts of atmospheric oxygen for growth
-excess oxygen blocks the activities of their oxidative enzymes and results in death

142
Q

require the absence of free oxygen for growth because the require the presence of molecules other than oxygen to act as the final electron acceptor
-presence of oxygen is lethal

A

obligate anaerobes

143
Q

fermentative organisms that do not use oxygen as a final electron acceptor
-produce enzyme so they can survive in the presence of toxic oxygen

A

aerotolerant anaerobes

144
Q

can grow in the presence or absence of free oxygen
-preferentially use oxygen for aerobic respiration but can perform cellular respiration anaerobically if necessary
-can use nitrates or sulfates as final hydrogen acceptors or use a fermentative pathway

A

facultative anaerobes

145
Q

proof of life/viable counts

A

show that an organism can replicate and form colonies on an agar plate

146
Q

serial 10 fold dilutions

A

allow estimation of the number of live organisms in the initial sample

147
Q

pros of serial dilution

A

allow for colony isolation, quantifies only live cells, controls for mixed cultures

148
Q

cons of serial dilution

A

takes longer (incubation time), user error, material use

149
Q

less than 30 colonies, TFTC

A

too few to count

150
Q

more than 300 colonies, TNTC

A

too numerous to count

151
Q

colony forming units / milliliter of initial culture
CFU/mL

A

how to present results of serial dilution viable counts

152
Q

CFU calculation formula

A

(number of colonies on plate) x (reciprocal of dilution of sample)

153
Q

standard CFU formula

A

(colony count on agar plate) / (total dilution of tube) x (amount plated)

154
Q

bacteria normally reproduce by __ __

A

binary fission- how bacteria reproduce

155
Q

intense activity preparing for population growth, but no increase in population

A

lag phase of the growth curve

156
Q

logarithmic, or exponential, increase in population

A

log phase of the growth curve

157
Q

period of equilibrium; microbial deaths balance production of new cells

A

stationary phase of the growth curve

158
Q

population is decreasing at a logarithmic rate

A

death phase of the growth curve

159
Q

the logarithmic growth in the log phase is due to reproduction by ___ ___ (bacteria) or __ (yeast)

A

binary fission, mitosis

160
Q

the generation time of a culture

A

the growth rate of a culture, determined by the number of cells at several time points, only calculated during the log phase

161
Q

equation of generation time

A

g = t/n
g - generation time (minutes)
t - time of exponential growth
n - the number of generations

162
Q

turbidity- optical density

A
  • 600 nm light source
    detects “cloudiness” or transmittance through a medium
163
Q

pros of optical density

A

quick and easy

164
Q

cons of optical density

A

do not distinguish between live and dead cells, contamination not taken into account, does not allow for isolation of colonies

165
Q

formula for n

A

(log N - log N0) / log 2

166
Q

axes of graph converting optical density to cell number (original graph)

A

x- optical density
y- cell number

167
Q

axes of graph used to determine generation time (new graph)

A

x- time
y- cell number

168
Q

polymer of collagen that makes up connective tissues, liquid above 25°C

A

gelatin

169
Q

break down collagen
-nutrient acquisition, virulence

A

gelatinases (collagenases)

170
Q

liquefaction after growth followed by refrigeration indicates hydrolysis

A

gelatin hydrolysis test

171
Q

iodine is used to detect the presence of starch, hydrolysis revealed as a clear zone around bacterial growth

A

amylase/starch hydrolysis test

172
Q

tributyrin agar, lipase-positive organisms produce clear zone around growth

A

lipid hydrolysis

173
Q

converts 1 molecule of glucose to 2 molecules of pyruvate

A

glycolysis
Embden-Meyerhof pathway

174
Q

molecular oxygen as the final electron acceptor (more ATP generated)

A

aerobic cellular respiration

175
Q

inorganic ions rather than oxygen are final electron acceptor

A

anaerobic cellular respiration

176
Q

doesn’t require oxygen and organic substrate is the final electron acceptor

A

fermentation

177
Q

may cause acid and gas

A

carbohydrate fermentation

178
Q

indicator of acid production (yellow = acid)

A

phenol red indicator

179
Q

indicator of presence of air bubble

A

durham tube

180
Q

indicates that an organism can metabolize sugar in the tube

A

color change red to yellow, acid production

181
Q

fermentation and gas production

A

color change and bubble in the durham tube

182
Q

color change to a dark pinkish-red for carbohydrate fermentation

A

indicates a basic or alkaline metabolic product due to utilization of the peptone rather than the sugar

183
Q

reacts with indole to produce red/pink color

A

Kovac’s reagent

184
Q

tryptophanase

A

enzyme used to identify bacteria that produce indole

185
Q

produced by certain enterobacteriaceae by two pathways

A

H2S (Hydrogen sulfide gas)

186
Q

combines with hydrogen sulfide gas to form black sulfide precipitate

A

FeSO4 (ferrous ammonium sulfate)

187
Q

a positive result for __ is indicated when radiating outward from the central stab; a negative result shows only along the stab line

A

motility

188
Q

all ferment glucose to organic acids (the acid varies)

A

enteric microorganisms

189
Q

differentiates between E. coli and K. aerogenes final end products

A

MR-VP tests

190
Q

determine ability to oxidize glucose to make acid end products (E. coli)- makes red color

A

methyl red test

191
Q

K. aerogenes converts acids to acetylmethylcarbinol raising the pH brining color back to yellow
-barritt’s reagent turns pink with acetylmethylcarbinol

A

voges proskauer test

192
Q

determine the ability of an organism to use the enzyme citrase to use citrate as a sole carbon source

A

citrate IMViC test

193
Q

growth on the slant and blue color

A

means citrate was utilized as a carbon source

194
Q

differentiate organisms based on their ability to hydrolyze urea with the enzyme urease

A

urease test

195
Q

urinary tract pathogens from the genus __ may be distinguished from other enteric bacteria by their rapid urease activity

A

Proteus

196
Q

broth tube turns bright pink

A

positive urease test

197
Q

fissures or cracks in the clot of litmus tube

A

gas production in litmus tube

198
Q

acid clots solidify the medium and can appear __ or __ with a pink band at the top depending on the oxidation-reduction status of litmus

A

pink, white

199
Q

lactose fermentation acidifies the medium and turns the litmus pink

A

pink in litmus milk test

200
Q

reduced litmus is white

A

white color in litmus test

201
Q

oxidized litmus is purple

A

purple in litmus test

202
Q

alkaline reaction for litmus test

A

blue/purple medium or blue band at the top

203
Q

stormy fermentation

A

heavy gas production that breaks up the clot

204
Q

digestion of peptone

A

milk protein completely digested, clear to brown fluid

205
Q

acid clot formation

A

proteolysis

206
Q

enzyme catalase

A

degrades hydrogen peroxide

207
Q

performed on a glass slide, colony + hydrogen peroxide - bubbles form

A

positive catalase test

208
Q

uses a chromogenic reducing agent as an indicator to detect bacteria that produce cytochrome oxidase

A

oxidase test

209
Q

indicator in oxidase test
- donates electrons to cytochrome oxidase, becomes self oxidized changing from light pink reduced to a dark maroon almost black oxidized compound

A

p-aminodimethylalanine oxalate

Pre-health Microbiology (5 decks)

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