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BIO 286 - Microbiology > Lab Practical Review (Bio 286 - Microbiology) > Flashcards

Lab Practical Review (Bio 286 - Microbiology) Flashcards

1
Q

types of pathogens used in the laboratory

A

BSL-1 (most common; minimal hazard; examples include Escherichia coli, Bacillus cereus, and Saccharomyces cerevisiae) or BSL-2 (less common; moderate hazard; example includes Staphylococcus aureus)

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2
Q

general procedures followed in laboratory

A

wear appropriate lab attire (including lab coat)… no eating/drinking in lab… report accidents or incidents to lab professor… behave appropriately… bench cleaning (beginning and end of lab session)… handwashing (after lab sessions)… clean up spills… proper waste disposal (non-contaminated trash into regular trash bins, glass waste into glass container, staining waste into stain receptacle, contaminated waste into red bag containers)

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3
Q

dangerous equipment used in laboratory

A

bunsen burner (most dangerous); bacterial stocks

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4
Q

ocular lenses

A

eye pieces of microscope; each has a magnifying power of 10X

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5
Q

cord wrap

A

brackets to store the power cord around when done using the scope

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6
Q

objective lenses

A

secondary lenses to magnify the sample; range from 4X to 100X

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7
Q

course focus knob

A

used to rapidly move the stage (a sample) to focus

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8
Q

fine focus knob

A

used to tweak the focus… used with 40X and 100X objectives

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9
Q

mechanical stage controls

A

move the sample in the X and Y planes, with the focus on the Z plane

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10
Q

condenser iris

A

lever controlling how much light passes through the sample

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11
Q

power switch

A

turns the lamp on and off

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12
Q

light source

A

halogen lamp

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13
Q

total magnification

A

ocular lens x objective lens

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14
Q

total magnification achieved by different objectives

A

objective 4X x ocular 10X = 40X…

objective 10X x ocular 10X = 100X…

objective 40X x ocular 10X = 400X…

objective 100X x ocular 10X = 1000X

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15
Q

immersion oil was used on the

A

100X objective

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16
Q

why is immersion oil used?

A

to keep light from bending… to reduce light loss between the slide and the lens… to improve the resolution

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17
Q

calibrate microscope using a stage micrometer

A

scale divisions x 10 / # ocular divisions

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18
Q

use the reticle to measure a cell size

A

after calibrating the microscope to determine how many micrometers each division of the ocular ruler is, count how many ocular lines it takes to span the bacterial cell

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19
Q

gram negative bacteria

A

pink/red

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20
Q

gram positive bacteria

A

purple

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21
Q

cocci

A

spherical bacteria

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22
Q

streptococci

A

spherical bacteria that form a chain

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23
Q

staphylococci

A

Spherical shaped bacteria that form grape-like clusters

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24
Q

bacilli

A

Rod shaped bacteria

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25
Q

coccobacilli

A

a bacterium that is an oval rod

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26
Q

streptobacilli

A

rod-shaped bacteria occurring in chains

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27
Q

spirochete

A

spiral shaped bacteria

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28
Q

firmicutes

A

bacteria with gram positive cell wall

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29
Q

proteobacteria

A

bacteria with gram negative cell wall

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30
Q

eukaryotes

A

bacteria with gram positive cell wall but irregular bacterial shape

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31
Q

gram stain primary stain

A

crystal violet

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32
Q

gram stain mordant

A

gram’s iodine

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33
Q

gram stain decolorizer

A

acetone alcohol

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34
Q

gram stain secondary stain

A

safranin

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35
Q

crystal violet

A

used as a primary basic dye which can be taken up by the majority of bacteria due to its ability to rapidly permeate all cell walls

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36
Q

gram’s iodine

A

used as mordant to “seal” the Crystal Violet stain into the cell walls of the bacteria

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37
Q

acetone alcohol

A

used in gram stain to wash away crystal violet from gram negative bacteria; used as a decolorizer

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38
Q

safranin

A

counterstain in gram stain; used as a secondary stain after the initial crystal violet is washed away

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39
Q

capsules

A

clear structures surrounding the cells

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40
Q

capsules

A

clear structures surrounding the cells… function is to evade phagocytosis… composed of polysaccharides

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41
Q

capsule stain

A

stain used to stain cells that form capsules

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42
Q

india ink

A

stain used to visualize the capsule by making the background of the slide dark

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43
Q

india ink

A

stain used to visualize the capsule by making the background of the slide dark

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44
Q

endospore

A

function as “survival” cells to withstand harsh conditions… appear as greenish blue structures

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45
Q

vegetative cell

A

a cell that has not formed spores or other resting stages… appear as pink structures

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46
Q

gram stain

A

crystal violet -> iodine -> acetone alcohol -> safranin

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47
Q

endospore stain

A

malachite green (primary stain) -> heat (mordant) -> water (decolorizer) -> safranin (secondary stain)

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48
Q

organisms forming endospores

A

microbes of the Firmicutes (primarily Bacillus and Clostridium)

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49
Q

more endospores were made when grown on TSA

A

because TSA is less nutrient rich than BHIA, causing the cells to need to form more endospores in response to starvation conditions

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50
Q

acid-fast stain

A

a differential stain used to visualize acid-fast bacteria

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51
Q

acid-fast primary stain

A

basic fuchsin (+ phenol mordant, performed simultaneously)

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52
Q

acid-fast secondary stain

A

methylene blue

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53
Q

two pathogens that would appear violet/red with acid fast stain

A

Mycobacterium tuberculosis and Mycobacterium leprae

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54
Q

mycolic acid

A

compound within the cell walls of acid-fast bacteria (Mycobacterium tuberculosis and Mycobacterium leprae) that makes them waxy

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55
Q

flagella

A

external bacterial structures that function by providing motility to the cell

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56
Q

monotrichous flagella

A

single flagellum at one end

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57
Q

amphitrichous flagella

A

flagella at both ends

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58
Q

peritrichous flagella

A

flagella that cover the surface of a cell

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59
Q

lophotrichous flagella

A

cluster of flagella at one end of the cell

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60
Q

flagella provide motility through

A

spinning (like an electric motor)…rapidly rotating and providing thrust

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61
Q

flagella in eukaryotes

A

provide motility through wave-like motion

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62
Q

entamoeba histolytica

A

Causative agent of amoebic dysentery– a severe diarrheal disease associated with ingestion of contaminated food or water – part of SARCODINA

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63
Q

balantidium coli

A

The only ciliate human pathogen… these cells are transmitted by fecal contamination – part of CILIOPHORA

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64
Q

trichomonas vaginalis

A

Causes vaginitis and is an example of a sexually transmitted protozoan; only has a trophozoite form – part of MASTIGOPHORA

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65
Q

giardia lamblia

A

has a much reduced (nearly absent) mitochondria… Causes diarrheal diseases – part of EXCAVATA

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66
Q

plasmodium falciparum

A

causes malaria… ring stage trophozoite can be seen in some red blood cells – part of APICOMPLEXANS

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67
Q

fusarium verticilliodes

A

filamentous fungus widely distributed on plants and in the soil… a member of the class ASCOMYCETES and has SEPTATE hyphae… “football-shaped” macroconidia…may cause various infections in humans–one of the emerging causes of opportunistic mycoses

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68
Q

penicillium chrysogenum

A

filamentous fungus that is widespread… it can be found in soil, decaying vegetation, and the air…. It is a member of the class ASCOMYCETES and has SEPTATE hyphae… The conidia form on finger-like projections… may cause infections, particularly in immunocompromised hosts, but also produces the ANTIBIOTIC PENICILLIN

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69
Q

rhizopus stolonifer

A

cosmopolitan filamentous fungus found in soil, decaying fruit and vegetables, animal feces, and old bread (BLACK BREAD MOLD)… It is a member of the class ZYGOMYCETES and has COENCYTIC hyphae… common contaminants, but can also occasional causes of serious (and often fatal) infections in humans

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70
Q

syncephalastrum racemosum

A

filamentous fungus that is commonly isolated from soil and animal feces particularly in tropical and subtropical areas… It is a member of the class ZYGOMYCETES and has COENCYTIC hyphae. This heterothallic fungus requires a mating strain to produce zygospores… commonly considered as a contaminant and is very rarely associated with human disease

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71
Q

tube dilution factor (TDF)

A

sample + diluent / sample

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72
Q

serial dilution factor (SDF)

A

TDFa x TDFb x TDFc … (product of all TDFs)

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73
Q

CFU/mL

A

CFU x SDF / volume (mL) plated

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74
Q

plaques

A

holes in the bacterial lawn… caused by viral propagation overwhelming the host cells

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75
Q

microbial interaction represented by plaques

A

parasitism

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76
Q

virus (bacteriophage, T4)

A

caused the formation of plaques

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77
Q

lag phase

A

“flat” period of adjustment, enlargement; little growth

78
Q

log phase

A

The period of exponential growth of bacterial population

79
Q

stationary phase

A

period of equilibrium; microbial deaths balance production of new cells

80
Q

death phase

A

population is decreasing at a logarithmic rate

81
Q

growth rate (g)

A

log(2)/m [m = slope]

82
Q

generation time (k)

A

1/g (x 60 if in hours)

83
Q

m

A

log(y1) - log(y2) / x1 - x2

84
Q

non-halotolerant

A

cannot withstand high salt concentrations

85
Q

halotolerant

A

a microorganism that does not require NaCl for growth but can grow in the presence of NaCl

86
Q

halophile

A

an organism that can grow in, or favors environments that have very high salt concentrations

87
Q

acidophile

A

an organism that grows best at low pH; typically below pH 6

88
Q

neutrophile

A

an organism that grows best at neutral pH, between pH 5.5 and 8

89
Q

alkalinophile

A

an organism that grows best at basic/alkaline pH, between pH 8 - 11

90
Q

psychrophile

A

bacteria that prefer cold, thriving at temperatures between 0 C and 25 C.

91
Q

mesophile

A

bacteria that prefers moderate temperature and develops best at temperatures between 25 C and 40 C

92
Q

thermophile

A

bacteria that thrive best at high temperatures, between 40 C and 70 C

93
Q

GasPak chamber

A

sodium citrate and sodium bicarbonate react to release carbon dioxide, and the activated charcoal in the packet binds free oxygen gas – removing oxygen and creating a carbon dioxide atmosphere, ie anaerobic conditions

94
Q

organisms that can grow in GasPak chamber

A

anaerobes, facultative anaerobes, aerotolerant anaerobes

95
Q

organisms that cannot grow in GasPak chamber

A

aerobes

96
Q

organisms that could grow in GasPak chamber or in air

A

facultative anaerobes, aerotolerant anaerobes

97
Q

methylene blue

A

oxygen indicator used in BAA plates

98
Q

resazurin (or methylene blue)

A

oxygen indicator used in fluid thioglycolate tubes

99
Q

aerobe

A

an organism that requires oxygen for respiration and can live only in the presence of oxygen

100
Q

aerotolerant

A

an organism that does not utilize oxygen but can survive and grow in its presence

101
Q

anaerobe

A

an organism that grows without air, or requires oxygen-free conditions to live

102
Q

microaerophilic

A

an organism that can require a lower oxygen concentration than what is in air

103
Q

facultative anaerobe

A

an organism that prefers an oxygen environment but is capable of living and growing in its absence

104
Q

zone of inhibition

A

region around a chemical saturated disc, where bacteria are unable to grow due to adverse effects of the compound in the disc

105
Q

most effective drug

A

forms the largest zone of inhibition

106
Q

least effective drug

A

forms the smallest/no zone of inhibition

107
Q

antiseptics

A

used on living surfaces (like skin); tend to be less toxic… include isopropanol (alcohol), iodine (oxidizer), dish soap (surfactant), and germicidal soap (phenolic)

108
Q

antibiotics

A

produced by other microorganisms and tend to be used chemotherapeutically… include peptidoglycan attackers (ampicillin, cephalothin, penicillin) and 70s ribosome attackers (erythromycin, streptomycin, tetracycline)

109
Q

disinfectants

A

used on inanimate objects and tend to be more toxic… include lysol (phenolic), roccal (quaternary ammonium compound), bleach (oxidizer), and aldehyde (alkylator)

110
Q

resistant bacteria

A

small/no zone of inhibition

111
Q

intermediate bacteria

A

intermediate sized zone of inhibition

112
Q

susceptible bacteria

A

large zone of inhibition

113
Q

thermal death time

A

the shortest amount of time needed to kill all microbes at a given temperature

114
Q

thermal death point

A

the lowest temperature needed to kill all microbes in a given time

115
Q

most heat-resistant bacteria

A

bacillus or clostridium because they form endospores

116
Q

fecal coliforms

A

lactose-fermenting gamma proteobacteria; typically found in mammalian feces… appears blue on mFC plate

117
Q

fecal streptococci

A

typically found in feces of animals (like ducks, not humans)… appears red on mEA plate

118
Q

human contamination source

A

blue (coliforms) / red (streptococci) > 4:1

119
Q

mixed contamination source

A

2:1 > blue (coliforms) / red (streptococci) < 4:1

120
Q

unknown contamination source

A

0.7:1 > blue (coliforms) / red (streptococci) < 2:1

121
Q

animal contamination source

A

blue (coliforms) / red (streptococci) < 0.7

122
Q

mFC plate

A

used to detect fecal coliforms… contains lactose and resorcinol dye that makes coliforms appear blue… contains bile salts that inhibit growth of non-coliforms

123
Q

mEA plate

A

used to detect fecal streptococci… contains metabolic poison sodium azide and tetrazolium red that makes streptococci appear red… sodium azide prevents growth of non-streptococci

124
Q

transformation

A

process in which exogenous DNA is incorporated into another cell’s genome

125
Q

in the transformation experiment, what microbe was used?

A

two strains of Acinetobacter calcoaceticus, one strain resistant to streptomycin and the other strain susceptible to streptomycin

126
Q

in the transformation experiment, what antibiotic was used?

A

streptomycin

127
Q

streptomycin targets

A

70S ribosome

128
Q

does streptomycin work against yeast?

A

no because yeast are eukaryotes that have 80S ribosomes

129
Q

why does UV kill?

A

it forms pyrimidine dimers, preventing proper base pairing in DNA… causing mutations in DNA sequence where enough mutations will kill the cell

130
Q

UV wavelength used in germicidal lamps

A

260 nm (UVC)

131
Q

where is UV light used at the wastewater treatment plant?

A

wastewater disinfection

132
Q

what organisms were the most resistant to UV killing?

A

Bacillus cereus (most resistant because it forms endospores) > pseudomonas aeruginosa (because the green pigment protects it) > serratia marcescens (because the red pigment protects it) > staphylococcus aureus (most susceptible)

133
Q

lichens exhibit

A

mutualism behaviour

134
Q

termites and their hindgut microbiota exhibit

A

mutualism behaviour

135
Q

cow and their ruminants exhibit

A

mutualism behaviour

136
Q

staphylococcus aureus and clostridium sporogenes exhibit

A

commensalism behaviour

137
Q

lactose fermentation (MacConkey plate) positive result

A

RED

138
Q

lactose fermentation (MacConkey plate) negative result

A

tan/brown

139
Q

citrate positive result

A

BLUE

140
Q

citrate negative result

A

green

141
Q

vogues proskauer positive result

A

RED

142
Q

vogues proskauer negative result

A

brown

143
Q

TSI H2S positive result

A

BLACK

144
Q

TSI H2S negative result

A

no change

145
Q

indole positive result

A

RED

146
Q

indole negative result

A

yellow

147
Q

urea positive result

A

PINK

148
Q

urea negative result

A

yellow

149
Q

MacConkey Agar

A

if the microbes ferment lactose then the acids they produce turn the indicator dyes red… a positive result is red and a negative result is no change

150
Q

indole

A

if microbes breakdown amino acid tryptophan to indole, the indole reacts with Kovac’s reagent to turn bright red… a positive result is red and a negative result is yellow

151
Q

Triple Sugar Iron Agar (slant)

A

if ammonia is produced then the pH of the media becomes neutral – red and if lactose/sucrose is fermented then the tube will remain yellow… a positive result is yellow (A) and a negative result is red (K)

152
Q

triple sugar iron agar (butt)

A

the butt of the TSI is always yellow – a positive result is yellow (A)

153
Q

triple sugar iron agar (gas)

A

gases produced during fermentation will appear as bubbles or cracks… a positive result is cracking and a negative result is no change/no cracking

154
Q

triple sugar iron agar (H2S)

A

hydrogen sulfide produced from breakdown of amino acid cysteine reacts with the iron to form ferric sulfide which is black… a positive result is black and a negative result is no change

155
Q

MRVP (methyl red)

A

turns red in the presence of organic acids… a positive result is red and a negative result is yellow

156
Q

MRVP (voges proskauer)

A

detects acetoin to turn red in its presence… a positive result is red and a negative result is coppery brown/no change

157
Q

citrate

A

at acidic pH (citrate present) the media is green and if the citrate is metabolized the pH will turn blue… a positive result is blue and a negative result is green

158
Q

NaCl broth positive result

A

GROWTH – CLOUDY

159
Q

NaCl broth positive result

A

GROWTH – CLOUDY

160
Q

NaCl broth negative result

A

no growth – clear

161
Q

bile esculin (BE) slant positive result

A

BLACK

162
Q

bile esculin (BE) slant negative result

A

tan

163
Q

optochin positive result

A

SUSCEPTIBLE

164
Q

optochin negative result

A

resistant

165
Q

DNase (HCl) production positive result

A

CLEARING

166
Q

DNase (HCl) production negative result

A

no clearing

167
Q

mannitol salt agar

A

selective and differential media; contains 6.5% NaCl, phenol red dye, and mannitol…. microbes that are able to grow may ferment mannitol or not (if they do, the acids they produce turn the indicator dyes in the media yellow)

168
Q

catalase

A

enzyme that decomposes hydrogen peroxide to water and oxygen gas– when H2O2 is placed on the microbes, the production of bubbles indicates a positive reaction

169
Q

EF broth

A

media containing a fermentable sugar (glucose) and an acid indicator (bromcresol purple)– glucose fermentation results in acid end products, which cause the media to turn yellow… the media also contains a metabolic poison sodium azide, allowing only azide-resistant microbes to grow

170
Q

NaCl broth

A

contains 6.5% NaCl, allowing only halotolerant microbes to grow and turn the media cloudy

171
Q

bile esculin slant

A

media used to detect the metabolism of a compound called esculin– if it is converted to esculetin, it will react with iron in the media and turn completely black… bile acids in this media inhibit the growth of most gram positive organisms

172
Q

DNase

A

breaks down DNA to nucleotides by adding HCl to the DNA plate after microbes have been cultured– the low pH causes DNA to precipitate within the media, turning the late cloudy. any areas where DNA has hydrolyzed will not precipitate, leaving clear areas around growth of positive cultures

173
Q

sheep blood agar

A

some bacteria break down blood cells using toxins called hemolysins… resulting in three patterns of hemolysis clearings- alpha, beta, and gamma… additionally, bacitracin and optochin are antibiotics used to determine susceptibility

174
Q

alpha hemolysis

A

partial breakdown of red blood cells– resulting in smoky green color (no longer red but no clearing)

175
Q

beta hemolysis

A

complete breakdown of red blood cells, resulting in complete clearing– can read text through the plates

176
Q

gamma hemolysis

A

no clearing, the plates have growth on them but are just as red

177
Q

bacitracin

A

an inhibitor of peptidoglycan biosynthesis that group A streptococci are particularly susceptible to

178
Q

optochin

A

a copper containing compound that pneumococci are particularly susceptible to

179
Q

amylase exoenzyme

A

exoenzyme used to break down polymer amylose (starch) [polysaccharide]… visualized by a clear zone around bacteria after addition of iodine… performed by bacillus cereus

180
Q

DNase exoenzyme

A

exoenzyme used to break down polymer DNA [polynucleotide]… visualized by clear zone around colonies after addition of HCl… performed by staphylococcus aureus

181
Q

gelatinase exoenzyme

A

exoenzyme used to break down polymer gelatin [protein]… visualized by gelatin not solidifying when on ice… performed by staphylococcus aureus, bacillus cereus, and pseudomonas aeruginosa

182
Q

lipase exoenzyme

A

exoenzyme used to break down polymer lipid (triglyceride) [lipid]… visualized by less opaque media and a halo around the bacterial growth… performed by pseudomonas aeruginosa and staphylococcus aureus

183
Q

lysozyme

A

targets peptidoglycan of bacteria… an important component of immune defense… found in eggs, tears, saliva, and can be commercially made

184
Q

E.coli is resistant to lysozyme

A

because it is gram negative and has less peptidoglycan in its cell wall

185
Q

M.luteus

A

gram positive bacterium that is a normal inhabitant of human microbiota

186
Q

neutrophil

A

most common white blood cell… main phagocytic cells in circulation

187
Q

lymphocyte

A

second most common white blood cell… includes B-cells and T-cells of adaptive immune response

188
Q

monocyte

A

immune cells that differentiate into tissue macrophage and dendritic cells… important antigen presenting cells

189
Q

eosinophil

A

cells filled with pink stained granules… deal with worm infections and are also associated with type I hypersensitivity (allergies)

190
Q

basophile

A

least common/rarest white blood cell… filled with purple stained granules… associated with anemia and some microbial infections

BIO 286 - Microbiology (21 decks)

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