Lab practical 1 Flashcards
(150 cards)
1
Q
What is appropriate PPE?
A
Gloves, lab coats, closed toe shoes, long hair tied
2
Q
Where do gloves and used plate cultures go for disposal?
A
Biohazard bag
3
Q
Moves stage up/down and side to side
A
Stage controls
4
Q
Moves stage closer/further from object lens
A
Course focus
5
Q
Brings specimens into sharp focus
A
Fine focus
6
Q
Focuses light
A
Condenser
7
Q
Varies the intensity and size of the cone of light
A
Iris
8
Q
Total magnification of light compound microscope =
A
Objective lens x eyepiece
9
Q
Used to describe the sharpness and detail of the image
A
Resolution
10
Q
Used to describe the enlargement of an object under the microscope
A
Magnification
11
Q
Sphere microorganisms
A
Coccus
12
Q
Rod microorganisms
A
Bacillus
13
Q
Spiral microorganisms
A
Spirillum
14
Q
Chain microorganisms
A
Strepto
15
Q
Cluster microorganisms
A
Staphylo
16
Q
Types of microscopy: light goes through the condenser
Dark object on light background
A
Bright field microscopy
17
Q
Types of microscopy: light object on a dark background
Unstained organisms
A
Dark field microscopy
18
Q
Types of microscopy: hollow cone of light
Improves contrast between object/background
A
Phase control microscopy
19
Q
Types of microscopy: bright object dark background
Chemiluminescence
Phosphorescence
A
Fluorescence microscopy
20
Q
Types of microscopy: electrons bounce off specimen
Visualization of the surface of specimen
Specimens coated in electron dense material
A
Scanning electron miscroscopy
21
Q
Types of microscopy: beam of electrons pass through the specimen
A
Transmission electron microscopy
22
Q
Multicellular fungi
Hyphae
A
Multicellular filamentous molds
23
Q
Portion that obtains nutrients
A
Vegetative hyphae
24
Q
Projects above the surface of the fungi, reproduction
A
Aerial hyphae
25
Incomplete budding of yeast cell
Pseudohyphae
26
Eukaryotes: fungi
Grow best at room temp and acidic ph 5.6
Fruiting bodies visible above ground
Macroscopic filamentous fungi
27
Eukaryotes: fungi
Single-celled
Multiple through budding
Sometimes opportunistic pathogens
Bread, brewing
Yeasts
28
Eukaryotes: algae
Unicellular, green
Photosynthetic
Found in temperature soils
Chlamydomonas
29
More closely related to eukaryotes than bacteria
Not pathogenic
Minority of exteemophiles
Archaea
30
Bacteria motility: corkscrew motion
Spirochetes
31
Bacteria motility: “run” and “tumble”
Flagellar
32
Bacteria motility: twitching/gliding motion
Pilli
33
Bacteria non-motile movement
Random
Collisions
Brownian movement
34
Staining: penetrates cell wall
Cell wall negatively charged
Positive charged any basic can be used (crystal violet, methylene blue)
Simple staining
35
Staining: stains background, not cell
Acidic dye, negatively charged
Negative staining
36
Staining: allows to differential between cell types
Gram staining
Differential staining
37
Gram stain: Purple stain
Gram positive
38
Gram stain: pink stain
Gram negative
39
Two extremes: oxygen is 100% intolerable, oxygen is 100% required
Obligate anaerobe or aerobe
40
Oxygen not necessary but can be tolerated
Oxygen may be beneficial
Facultative anaerobe
41
Cannot use oxygen
Oxygen not harmful
Aerotolerant anaerobes
42
Need oxygen but at lower levels
Microaerophiles
43
Kills microbes from tools, media, tables, other surfaces, gloves, etc.
Sterilization
44
Methods of sterilization
Autoclaves (moist heat)
Oven (dry heat)
Filtration
Radiation (UV and gamma)
45
Types of media: allows for fast growth
High density of microbes
Liquid (broth)
46
Types of media: used to check for motility
Anaerobic growth
Semi-solid
47
Types of media: used for colony isolation
Solid (agar)
Any broth can be made solid
48
Types of media: selects for certain types of bacteria
Selective media
49
Types of media: differentiates between bacteria
Differential media
50
Plating techniques: plate liquid sample containing bacteria
Can determine amount of bacterial cells in original liquid culture
Spread plate
51
Micropipette ranges: 0.1-2.0
P2
52
Micropipette ranges: 0.5-20.0
P20
53
Micropipette ranges: 20.0-200.0
P200
54
Micropipette ranges: 100-1000
P1000
55
Counts love and dead to get total number
Direct counting
56
Only counts living
Viable counting
57
Viable counting range
(TFTC) 30-300 (TNTC)
58
Light is inversely proportional to number of cells
Spectrophotometer
Turbidity
59
Dilute culture in stepwise manner
Serial dilution
60
Measures light absorbance at specific wavelength
Spectrophotometer
61
Phases of bacterial growth: No cell division
Lag phase
62
Phases of bacterial growth: generation time constant in
Exponential growth phase
63
Phases of bacterial growth: reproduction and death are balanced in
Stationary phase
64
Phases of bacterial growth: death rate exceeds reproduction in
Death phase
65
Common storage methods for culture samples
Agar slants
Permanents
Freezing cultures
Lyophilized cultures
66
Recommended storage time: less than 2 weeks
Agar plates
67
Recommended storage time: 3weeks to 6 months
Agar slants
68
Recommended storage time: 3 weeks to 12 months
Stored at room temp or cooler
Stab culture
69
Long term storage 1-3 years
-20C or -80C
Freezing cultures
70
Long term storage 15+ years
Culture is dried and frozen
Lyophilization
71
Common stressors
Temperature
pH
Osmotic pressure
Radiation exposure
72
Temperature classifications: 0-20C
Psychrophiles
73
Temperature classifications: 20-45C
Mesophiles
74
Temperature classifications: 45-80C
Thermophiles
75
Temperature classifications: 75-130C
Hyperthermophiles
76
pH classifications: pH 0 to 5.5
Vinegar, lemon juice
Acidophiles
77
pH classifications: pH 5.5 to 8
Water, blood
Neutrophiles
78
pH classifications: pH 8 to 14
Bleach, drain cleaner
Alkalphiles
79
Osmotic pressure classifications: low (solute) outside cell compared to inside
Hypotonic
80
Osmotic pressure classifications: high (solute) outside cell compared to inside
Hypertonic
81
Osmotic pressure classifications: equal (solute) inside/outside
Isotonic
82
DNA repair: need light for activation
Photolyase
Repairs thymine dimers
Less likely to cause mutations
Photoreactivation
83
DNA repair: recombination repair
DNA polymerase repairs new gap
Prone to mutations
RecA
84
Several different species occupying the same area
Community
85
Amplifies known sequence of DNA via ‘primers’
Applications: sequencing, molecular cloning, forensics, diagnostics (COVID)
Polymerase chain reaction
86
Derived from a thermophilic bacterium
Thermostable up to 95C
Taq polymerase
87
Utilizes temperatures and duration that are programmed into the machine
Thermocycler
88
Control that isn’t expected to give result
Negative control
89
Control that is expected to give result
Positive control
90
Highly concerved
All microbes will have the same NT sequence
Positive control
fis
91
Variable gene
Different microbes have different NT sequence for this gene
Allow us to identify the organism
dps
92
Relative ability of microbes to survive and grow in a particular environment
Fitness
93
Can evolve to increase their fitness:
Horizontal gene transfer
Recombination
Mutation
94
Separates nucleic acids/proteins based on size
Molecules are mobilized in an electric field - RUN TO RED
Gel electrophoresis
95
Made of agarose and buffer
Increased % is better for smaller molecules
Decreased % is better for larger molecules
Agarose gel
96
Adds weight to the DNA to keep it in the well
Easier to see
Loading dye
97
Contains fragments of known sizes
DNA ladder
98
Binds to DNA and allows visualization once placed on blue light
Gel stain
99
Soil Horizons: O =
Humus layer
100
Soil Horizons: A =
Topsoil
101
Soil Horizons: B =
Subsoil
102
Soil Horizons: C =
Regolith
103
Soil Horizons: R=
Unweathered bedrock
104
What have the largest biomass
Fungi
105
What produce around 75% of natural antibiotics
Actinomycetes
106
Secondary metabolites
Antibiotics
107
Antibiotics are not effective against
Fungi
108
The buildup and breakdown of nutrients within a cell
Reactions catalyzed by enzymes
Metabolism
109
Binding sites specific for substrate
Typically proteins
Enzymes
110
Biochemical reaction: Breakdown of large molecules that not smaller, more simple molecules
Makes energy
Catabolic
111
Biochemical reaction: takes smaller molecules and builds them into larger molecules
Uses energy
Anabolic
112
Building blocs of proteins
Some are essential and must be obtained externally
Amino acids
113
Organic molecules that contain carbon, hydrogen, oxygen, and nitrogen
Inactive without the correct shape
Proteins
114
Protein structure: a chain of AAs (polypeptide chain)
Primary structure
115
Protein structure: repetitive twisting/folding of the polypeptide chain
Creates two different structures: alpha helixes or beta sheets
Secondary structure
116
Protein structure: alpha helices or beta sheets fold into a 3D structure
Tertiary structure
117
Protein structure: multiple, folded polypeptide chains coming together to form a protein
Quaternary structure
118
SIMs test: production of hydrogen sulfide
Sulfur
119
SIMs test: hydrolysis of tryptophan
Indole
120
SIMs test: motile bacteria
Motility
121
IMVic test: hydrolysis of tryptophan
Indole
122
IMVic test: organic acid production
Methyl red
123
IMVic test: presence of acetoin
Voges proskauer
124
IMVic test: citrate as a carbon source
Citrate
125
Metabolic test: Multiple sugars, fermentation
Glucose, lactose, fructose
Sugar fermentation test
126
Metabolic test: indicates if microbe can ferment lactose
Beta-galactosidase
127
Metabolic test: breakdown of starch to glucose
Carbon source
Starch hydrolysis
128
Test the microbe’s ability to complete sulfur reaction
Most useful for determining presence/absence of enteric bacteria within a culture
SIMs test
129
Test if the microbe can utilize tryptophan as a carbon and nitrogen source vi tryptophase
SIMs test
130
Semisolid media used in this test allows for the identification of motile bacteria
SIMs test
131
Can only be used on gram negative bacteria
Determines if the microbe can use citrate as a carbon source
IMViC test
132
What are the types of microscopy
Bright field
Dark field
Phase control
SEM
TEM
133
Parts of the compound light microscope
Eyepiece
Observation tube
Neck
Coaxial stage controls
Course focus
Fine focus
Nosepiece
Objective lens
Condenser
Iris
Light source
134
What tool do you use to measure microbes
Spread plate or pipette?
135
What are the types of microbial motility
Flagellar
Pilli
Spirochetes
136
Why do we stack bacteria
Helps with differentiating microorganisms
137
What is a smear preparation
Spreading bacteria across a glass slide
138
What is heat fixing
Toughens structures for staining and inactivates enzymes
139
Compare and contrast simple staining to differential staining
Simple: only one stain used
Differential: multiple stains used
140
What are the three domains of life
Eukarya
Bacteria
Archaea
141
CFU/ml =
Number of colonies per ml plated / total dilution factor
142
What are thermoduric bacteria
Heat resistant bacteria that can survive pasteurization
143
Can tolerate high concentrations of salt
Halotolerant
144
How can UV radiation affect microbes
Stop them from repairing themselvez
145
What enzymes repair DNA damage
DNA polymerase
146
What is the SOS response and when does it occur
Stops DNA synthesis when damage is too great
147
What are taxa
Communities that consist of two or more different species
148
What does does PCR stand for
Polymerase chain reaction
149
Why do we use PCR
Able to obtain billions of copies of DNA quickly
150
What produces antibiotics
Soil microorganisms
