Midterm Flashcards
1
Q
what kind of lab is Microbio 2120?
A
BSL-2 lab
2
Q
what is the guesting policy?
A
if a lab is missed you can guest in another lab that week
3
Q
what do you need to do in order to be allowed to guest?
A
- Notify by email
-YOU’RE TA
-guesting lab TA
-cc Knapp
4
Q
how much is each lab exercise turned in worth?
A
10 pts
5
Q
How much is each lab canvas quiz worth?
A
10 pts
6
Q
how many points do you get for attendance?
A
2 pts
7
Q
how many points is the unknown project worth?
A
85 pts
8
Q
how many points is the midterm worth?
A
100 pts
9
Q
how many points is the final worth?
A
200 pts
10
Q
where are slides disposed of?
A
place in cans on the ends on the end of benches
11
Q
where are plates & gloves disposed of?
A
placed in a biohazard waste container
12
Q
where do you dispose of plastic pipettes?
A
biohazard waste container
13
Q
where are test tubes disposed of?
A
place upright in racks in the test tube disposal tub on the counter in room 212
14
Q
where do you dispose of broken glass?
A
place in the glass waste disposal
15
Q
where do you dispose of the paper waste?
A
in regular
if used to clean up throw in the biohazard
16
Q
what kind of microbes does BSL-2 use?
A
use microbes that pose moderate hazards to laboratorians and the environment
17
Q
what are the biosafety levels?
A
Low risk:
BSL 1
BSL-2
BSL-3
BSL-4
High-risk microbes
18
Q
what the BSL-2 laboratories requirements
A
- access to the laboratory is restricted when work is being done
- lab coats and gloves, eye goggles, and face shields as needed
- procedures that can cause infection from aerosols or splashes are performed in a Biological safety cabinet
- an autoclave or an alternative method of decontamination is available or proper disposals
- lab is self-closing
- eyewash and sink
19
Q
what is the purpose of lab 1?
A
- to show the ubiquity of microorganisms
- to view patterns of bacterial growth on different forms of growth media: plates, slants, broth
- to understand the purposes for each media growth media type
20
Q
what types of microorganisms will likely be isolated on the plates?
A
bacteria found in dust particles
fungi in the air
21
Q
what types of microorganisms will NOT likely be isolated on the plates?
A
protozoan types
22
Q
what are the steps to lab 1:
A
- Obtain 6 plates and label them on the bottom (side with the agar) with your name, time, and the surface you will sample.
- Obtain one sterile swab.
- To obtain a sample, roll the sterile swab back and forth across the area you wish to sample.
- To inoculate the agar, roll the swab gently across the surface of the agar plate. Do not break the surface of the agar.
- When you are finished, dispose of the swab in the biohazard container and place the inoculated plates 1 & 2 at 25 C upside down & 3-6 in the 37°C incubator.
- Incubate the plates for 48 hours.
- After incubation, observe the plates for microbial growth
23
Q
what is used to describe elevation colony?
A
convex
umbonate
plateau
flat
raised
raised(spreading edge)
flat raised margin
growth into the medium
24
Q
what is used to describe the margin of a colony?
A
smooth entire
rhizoid
irregular lobate
filamentous
25
what is used to describe the whole colony?
round
irregular
filamentous
rhizoid
26
describe the growth in nutrient broth culture: Enterobacter aerogens
motile
produce uniform fine tumidity
27
describe the growth in nutrient broth culture: staphylococcus epidermis
non-motile
sediment
28
describe the growth in nutrient broth culture: mycobacterium smegatis
non-motile
waxy layer of bacteria leads to pellicle formation at the surface
29
what does flocculent mean?
suspended chunks or pieces
30
what does sediment mean?
growth on the bottom
31
what does ring growth mean?
growth at the top around the edge
32
what does pellicle mean?
membrane at the top
33
what is uniform turbidity?
evenly cloudy throughout
34
lab 1
-plate 1: lab air @25C
-plate 2: lab work bench @ 25C
-plate 3: lab work bench @37C
-plate 4: cough/sputum @37C
-plate 5: fingertips @37C
-plate 6: scalp/dander @37C
- anything at 37C would grow the most -> body temp
35
what factors would (or would not) influence growth and how the growth might appear on solid media
-warmth, moisture, pH, and O2
-colonies form
36
what is slant media used for?
-long term maintenance of isolates
37
what is liquid media used for?
-for growth studies or biochem analysis
38
what is solid media is used for?
obtain colonies
39
describe mycobacterium smegmatis:
fried egg appearance due to waxy cell membrane (mycolic acids)
40
describe streptomyces griseus:
-dry texture, has mold-like appearance
41
what happened to the proteus mirablilis?
-colonies spread(highly motile microbe)
42
what happened to the serrate marscens ?
-formed red colored pigment (when grown below 30C)
43
describe bacillus subtilis:
-solid odor; endospore forming bacteria
44
describe pseudomonas aeruginosa
-produce pigments; blue-green (pyocyanin) and yellow-green (pyoverdine)
45
what is defined media?
exact chem comp is known
46
what is complex media?
-contains one or more complex nutrients of undefined comp -> yeast extract; or enzymatic digests of beef or plant
47
what is pure cultures?
isolated bacterial species from environmental sample of mixed culture
48
what is the aseptic technique?
wire loop in fire
49
what is staining used for?
enhances contrast and resolution;
reveals details of cellular morphology and arrangement
-stain binds to different cell structures/molecules
50
what are the acidic dyes?
-nigrosin
-background of cell is stained
-negative chromatophores
51
basic dyes:
-crystal violet
-cell is stained
- positive chromatophores
52
-cocci
spherical
53
spirillum
spiral
54
bacillus
oval shaped
55
vibro
banna shaped
56
spirochete
spiral shape
57
pleomorphic
odd shaped
58
di-
pair
59
strepto-
chain
60
sarcinae
cube
61
staphylococci
grape like clusters
62
coccobacillus
short and plum
63
what does a heat fix do?
dries out cell wall and opens cells to dyes
64
simple stain:
-highlights cells to observe morphology and structures
-only uses one staining agent
-heat fixing
-basic dye: positively charged -> attracted to negatively charged cells
65
simple stain types
crystal violet, safranin, methylene blue
66
negative staining types
nigrosine, eosin
67
When does one use negative staining?
How is this method performed?
With what kind of stain?
-determine morphology and cellular arrangement
-for cells sensitive to heat-fixation during smear prep
-minimal distortion of cells
-two plate smear technique
68
differential staining
-allows for detection of differences between bacteria and serves as aid to ID bacteria
68
what the the types of differential staining
gram stain, acid-fast stain, spore stain, capsule stain
69
primary stain
stains gram postive
70
type of primary stains
crystal violet
71
counter stain
stains gram negative
72
what are the counter stain types?
safranin
73
what is the decolorizing agent?
alcohol
74
what does the decolonizing agent do?
washes crystal violet from gram negative
75
what does the mordant do?
binds crystal violet to gram positive
76
what are the types of mordant?
iodine
77
gram-postive
higher content of peptidoglycan and cross-linkage;
allows for retention of primary stain and resists decolorization
78
gram-negative
-less peptidoglycan, higher lipid content;
decolorization w/alcohol extracts lipids increasing porousness of cell;
results in loss of primary stain
79
endospores
A resistant, dormant structure formed inside of some bacteria that can withstand adverse conditions
80
acid fast
-carbolfuchsin stain for 5 min
-rinse with H2O and acid-alcohol
-counterstain with brilliant green
-rise with H2O and blot
81
acid fast appears
pink
82
nonacid fast appears
blue
83
describe nonacid fast cell envelope
· Cell envelope of acid‐fast bacteria contains mycolic acids (waxy)
84
PEA growth:
-organism is not inhibited by phenylethyl
85
PEA no growth
-organism is inhibited by PEA
86
growth with clearing CNA
-organism is not inhibited by CNA and completely hemolyzes RBCs
87
growth with greening of medium CNA
-organism is not inhibited by CNA and partially hemolyzes RBCs
88
growth with no change of medium color CNA
organism is not inhibited by CNA and does not hemolyze RBCs
89
no growth CNA
-organism is inhibited by CNA
90
no growth MSA
organism inhibited by NaCl
91
yellow growth or halo means
-organism not inhibited MSA and produced acid from mannitol ferm
92
red growth (no halo) means
-organism not inhibited by MSA and does not ferment mannitol, no reaction
93
no growth MAC
organism inhibited by crystal violet or bile
94
pink to red growth with or without bile precipitate MAC
organism is not inhibited and produces acid from lactose ferm
95
growth is "colorless" (not red or pink) MAC
organism not inhibited by crystal violet and does not ferment lactose, no reaction
96
growth is pink and mucoid EMB
organism is not inhibited by EMB and ferments lactose with little acid production
97
no growth EMB
growth is pink and mucoid EMB
98
Growth is dark (purple to black, with or without green metallic sheen) EMB
organism not inhibited by EMB and ferments lactose and/or sucrose with acid production
99
growth is "colorless" (no pink, purple, or metallic sheen) EMB
organism not inhibited by EMB and does not ferment lactose or sucrose, no reaction
100
-no growth HEA
-organism is inhibited by bile and/or one of the dyes
-gram positive
101
pink to orange growth HEA
-gram positive
-organism is not inhibited by bile and/or one of the dyes
- produces acid from lactose ferm
102
blue green growth with black precipitate HEA
-gram positive
-organism is not inhibited by bile and/or one of the dyes
-organism does not ferment lactose, but reduces sulfur to H2S
103
blue green growth without black precipitate HEA
-gram positive
-organism is not inhibited by bile and/or one of the dyes
-does not ferment lactose or reduce sulfur, no reaction
104
-reaction is gram positive:
PEA & CNA
105
reaction is gram negative:
HEA, EMB, MAC
106
MSA
selective for gram positive-high salt favors staphylococcus sp.
107
CNA
-selective for gram positive
-differential for hemolytic
108
MAC
-differential and selective
-inhibits gram positive ( via bile salts) and distinguishes between lactose (purple) and non lactose (colorless) fermenters
109
EMB
-differential and selective
-contains dyes eosin and methylene blue that inhibit gram positive
110
HEA
-selective and differential
-salmonella and shigella
-inhibitors of gram positive
111
condenser
focuses light through the specimen
112
objective lens
enlarges the image of the specimen
113
ocular lens
10X
114
light intensity knob
controls intensity of light
115
fine focus knob
used for precision focusing
116
coarse focus knob
used to focus the image on low power
117
stage
Supports the slide
118
iris diaphragm
controls the amount of light passing through the specimen
119
base of the microscope
supports the microscope
120
lamp of microscope
light source
121
parfocal
objective that can be changed with minimal or no refocusing
122
resolution
clarity/ sharpness of image
123
numerical aperture
light gathering ability of the objective lens and condenser
124
light wavelength
the distance between peaks in light waves
125
oil immersion
-used for observation of bacterial cells
126
limit of resolution
-minimum distance two points can be to still be viewed as separate points (resolving power)
127
total magnification:
-objective x ocular lens
128
scanning lens
X4
129
low power lens
X10
130
high-dry lens
X40
131
oil immersion lens
X100
132
transport of microscope
carry with two hands, one on arm of microscope, one under base;
place gently on table
133
cleaning of microscope
use only lens paper to clean lenses, including condenser, not paper towels
134
operation of microscope
-raise condenser to highest position; open iris diaphragm
-turn microscope lamp on; adjust to max intensity
-move the scanning (X4) or low power (X10) objective lens into place
-place slide on stage; center specimen over opening
-adjust iris diaphragm to yield optimum illumination
-to focus image, first coarse focus, then fine focus
135
prokaryote
-chromosomes are nonmembrane bound nucleoid region
-size: 1-10 um diameter
-lack organelles; has cytoskeleton elements
-domains eubacteria and archaea
136
eukaryote
-chromosomes located in membrane bound nucleus
-size: 10-100 um diameter
-organelles of specific form and function
-possess fully developed cytoskeleton
-domain eukarya
137
bacteria
prokaryotic
138
protozoans
eukaryotic
139
algae
eukaryotic
140
molds/yeast
eukaryotic
141
procedure for pure colonies
-dilution streaking method: plate is "seeded" with cells;
mechanical dilution of cells as they are spread on plate surface
-grows visible colonies
142
