Lab Practicum 3 Flashcards
(290 cards)
1
Q
What is the morphology description of a: coccus, bacillus, spirillum, vibrio, and spirochete
A
Coccus: spherical
Bacillus: rod
Spirillum: rigid spirals
Vibrio: comma
Spirochete: flexible spiral
2
Q
What is the arrangement description of a: diplo, staphylo, and strepto
A
Diplo: two
Staphylo: grape like clusters
Strepto: chains
3
Q
What arrangement and morphology pair does not exist
A
staphylobacillus
4
Q
What is the purpose of the capsule stain?
A
To differentiate between capsule producing cells and unencapsulated cell
5
Q
What is the function of the capsule
A
Acts as an adherence factor
Protects the bacteria from dehydration, nutrient loss, and phagocytosis (anti phagocytic)
6
Q
What is the composition of a capsule?
A
Mucoid polysaccharide (carbohydrate)
OR polypeptide
7
Q
What are you looking at when doing a capsule stain and what dyes are used?
A
Clear area = capsule
Cell
Basic stain: safranin
Background
Negative stain (acidic): congo red
8
Q
What is a sample organism that you are looking at when doing a capsule stain?
A
Klebsiella pneumoniae - causes pneumonia
9
Q
What is the purpose of a gram stain?
A
To stain and differentiate gram + bacteria from gram - bacteria
10
Q
What dyes do we use to stain a gram stain and what do they stain?
A
Basic dyes
Crystal violet: gram positive - purple
Safranin: gram negative - pinkish red
11
Q
Why do gram positive stain differently than gram negative
A
Gram positive have a thick layer of peptidoglycan which helps to retain the crystal violet
Gram negative have a thin layer of peptidoglycan that loses the crystal violet during decolorization
Secondary stain will make appearance pink red with safranin
12
Q
What tool do we use to transfer bacteria to slide when doing a heat fix and why?
A
We use a needle
We do not want too much bacteria
13
Q
Why do we heat fix?
A
Kill the cells
Adheres bacteria to the slide
14
Q
What is the mechanism behind heat fixing?
A
Proteins denature and coagulate
15
Q
What is the procedure of the gram stain
A
1) Primary stain: crystal violet for 1 minute and rinse
2) Mordant: iodine for 1 minute and rinse
3) Decolorization: acetone/alcohol until solution runs clear and rinse
4) Secondary stain (counterstain): safranin for 1 minute and then rinse
5) use kim wipe to blot off top (you can wipe the bottom)
16
Q
What is the most important step in the gram stain?
A
Decolorization
17
Q
What is the purpose of the iodine?
A
Creates a crystal violet iodine complex that makes the dye molecules larger so that crystal violet is retained in the cell of the gram positive bacteria
18
Q
What would the cells look like if you over or under decolorized?
A
Over: both will appear pinkish red as crystal violet will not be retained in either
Under: both will be purple from crystal violet being retained in both
19
Q
What is an example of a gram negative rod?
A
Escherichia coli
20
Q
What is an example of a gram positive cocci?
A
Staphylococcus aureus
21
Q
What is the purpose of the endospore stain?
A
To stain and visualize endospores and differentiate spore forming cells from non-spore forming cells (a differential stain)
22
Q
When will endospores form?
A
When conditions become harsh:
Temperature
pH
Dehydration
UV
Nutrient loss
Oxygen concentrations
23
Q
What is the endospore made of
A
A tough keratin protein
Makes endospore very resistant
24
Q
What is an example of an endospore producing bacteria?
A
Bacillus anthracis: causes anthrax
25
What genus are most common in producing endospores
Bacillus - aerobic
Clostridoum - anaerobic
26
Are most endospore producing bacteria gram + or gram -
Gram +
27
What dye is used to stain the endospore and what is the procedure when doing this?
Malachite green
We steam to help get the dye into the tough keratin protein
28
What is the counter stain in the endospore stain?
Basic dye - safranin
29
What is the end result of an endospore stain for a spore producing and non spore producing bacteria
Spore producing: red orange vegetative cell and green endospore in middle
Nonspore producing: red orange ONLY vegetative cell
30
What is the purpose of an acid fast stain?
To stain acid fast bacterial cell walls and differentiate acid fast bacteria from non acid fast bacteria
31
What is mycobacterium cell wall made of
Has a cell wall made of 60 percent mycolic acid
32
What is the purpose of having a mycolic acid cell wall
Adherence factor:
sticky/waxy to protect from:
Phagocytic digestion
Dehydration
Chemicals (antibiotics/disinfectants)
33
What dye do you need to steam when doing the acid fast stain and why?
Carbolfuchsin - to penetrate mycolic acid cell wall
34
What is the counter stain in the acid fast stain
Methylene blue
35
What color are acid fast bacteria and acid fast negative bacteria
Mycobacterium - pink
Non acid fast bacteria - blue
36
What is an example organism for an acid fast stain
Mycobacterium tuberculosis
Causes lung tb
37
What is the purpose of the phenyl red test?
To determine if bacteria can metabolize a specific carbohydrate
38
What is the purpose of the durham tube in the phenyl red test?
To detect the production of gas
39
Why are peptones included in the phenol red broth?
Food for general growth
40
Why is sodium chloride included in the phenol red broth?
Salt balance
41
Why is carbohydrate included in the phenol red broth
Substrate for metabolism of carbohydrate
Glucose
Or lactose
42
Why is phenol red included in the phenol red broth
Ph indicator
Yellow = acidic
red/orange = neutral
Pink = alkaline
43
What is the enzyme in the phenol red test?
Various endoenzymes
44
What is the product in the phenol red test?
Acids, alcohols, +/- gases
45
What does a yellow tube with turbidity mean for the phenol red test
Positive metabolism of sugar (with or without gas)
46
What does a red/pink tube with turbidity mean for the phenol red test
Negative metabolism of sugar (used peptones as food)
47
What does no growth in the tube mean for the phenol red test
No result (inconclusive)
48
Why must the readout be done between 24 and 48 hours after inoculation in the phenol red test?
Bacteria metabolize sugar first and lowers the pH in the first 24 hours
After sugar runs out, bacteria start to metabolize proteins and produce alkaline products and pH turns towards orange (or pink)
49
What does it mean to say bacteria are non-saccharolytic
Bacteria does not metabolize sugar
Appearance will be alkaline (pink) for both glucose and lactose
50
What is the purpose of the OF test?
Determining whether bacteria oxidize and/or ferment glucose
51
How do you inoculate the media for the OF test?
Stab inoculate the agar deep
52
Why do we use mineral oil in one of the tubes for the OF test
To seal off air for the anaerobic tube testing for fermentation
53
What buffer is used and what is the purpose of the buffer in the OF test
Dipotassium phosphate - keep the pH constant
54
What pH indicator was used in the OF test
Bromothymol blue
Yellow = acidic
Green = neutral
Blue = alkaline
55
What is the substrate in the OF test
Glucose
56
What is the enzyme in the OF test
Various endoenzymes
57
What is the product in the OF test
Aerobic tube (no mineral oil) - acids
Anaerobic tube (mineral oil) - acids, alcohol, +/- gases
58
What does a yellow on top and green on the bottom but anaerobic tube is green throughout mean for the OF test
Positive for oxidation and negative for fermentation
Strict oxidizer of glucose
59
What does yellow throughout mean in both tubes for the OF test
Positive for oxidation and negative for fermentation
Oxidizer and fermenter of glucose
Facultative anaerobe: grow with or without oxygen, but grows better with
60
What does blue on top but green on the bottom in the aerobic tube and green throughout on the anaerobic tube mean for the OF test
Negative for oxidation and negative for fermentation
Nonoxidizer and nonfermenter of glucose
Non saccharolytic: doesn’t use sugar, but uses peptones aerobically
61
How do we inoculate the citrate slant
Use a needle
Inoculate using a very light inoculum
Perform a zig zag inoculation
62
What tool do we use to inoculate the MRVP and Indole broths
Loop
63
What does each letter in the IMViC stand for
I: indole
M: methyl red
V: voges proskauer
C: citrate
64
What is the name of the media for the three tubes used in the IMViC test
MRVP broth
Tryptone broth
Citrate slant
65
What is the purpose of the indole test?
To determine whether bacteria can produce indole from tryptophan (an amino acid)
66
What is the substrate used in the indole test
Tryptophan
67
What is the enzyme used in the indole test
Tryptophanase
68
What is the product in the indole test
Indole, ammonia, and pyruvate
69
Why can’t you use a pH indicator as a readout for the indole test
Ammonia is a base and pyruvate is an acid which cancel each other out
70
What reagent do we use in the indole test
Kovac’s reagent - detects the presence of indole
71
What does a positive and negative look like for the indole test
Positive: turns pink (indole is present)
Negative: stays yellow (no indole produced)
72
What is the purpose of the methyl red and vogues proskauer test
To determine whether bacteria ferment glucose into mixed acids or acetoin
73
What is the substrate used in MRVP
Glucose
74
What is the enzyme used in MRVP
Various endoenzymes
75
What is the product for the methyl red test
Mixed acids (ph 4.4)
Decrease the pH
76
What is the product for the voges proskauer test
Acids (24 hours)
Acetoin (48 hours)
Increase the pH
77
What is used to differentiate in the MR test
Methyl red reagent (pH indicator)
Turns pink/red if the pH decreases (positive)
78
What is used to differentiate in the VP test
VPA and VPB (reagent)
Turns pink/red if acetoin is present (positive)
79
Why can’t the readout for VP be done before 24 hours
Produces acids initially and lowers the pH then produces acetoin, which raises the pH
80
Why are organisms that are MR+ usually VP-, and VP+ organisms usually MR-
If an organism is VP+, then it means that they produce acetoin, which raises the pH. Therefore, it shouldn’t test positive for MR too (which detects a low pH)
81
What does a positive and negative look like in the MR test
Positive: pink red - acids produced
pH less than 4.4
Negative: not pink red = acids not produced
Ph greater than 4.4
82
What does a positive and negative look like in the VP test
Positive: pink red - acetoin detected
negative : not pink red - acetoin not detected
Color at top for a negative could be a result of reagents reacting to one another
83
What is the purpose of the citrate utilization test
To determine whether bacteria can utilize citrate as a sole carbon source
84
Why do we add citrate to the citrate utilization test
Sole source of carbon - organisms need carbon to make ALL macromolecules
85
Why is ammonium phosphate included in the citrate utilization test
Source of nitrogen; if organism can use citrate as sole carbon source, it will break down the ammonium phosphate into basic products
86
What is the pH indicator used in the citrate test
Bromothymol blue
Yellow = acidic
green = neutral
Blue = alkaline
87
What is the substrate for the citrate test
Two substrates
Citrate
Ammonium phosphate
88
What is the enzyme used in the citrate test
Two endoenzymes
Citrate permease and citrate lyase
will perform citrate fermentation using citrate lyase and will grow on the agar
89
What is the product of citrate test
Ammonia + ammonia hydroxide
90
What is a positive and negative of the citrate test
Positive: blue (even a small amount) OR green with growth (bacteria ONLY grows if they can use citrate)
Negative: green with NO growth (bacteria ONLY grows if they can use citrate)
91
What is the purpose of the decarboxylase test
To determine whether bacteria can decarboxylase lysine or ornithine
92
Why is glucose included in the recipe for the decarboxylase test
For fermentation (bacteria use glucose 1st (at a low concentration) to lower the pH)
93
What is the pH indicator for the decarboxylase test
Bromocresol purple
Yellow = acidic
Orange = neutral
Purple = alkaline
94
Why is pyridoxal included in the decarboxylase recipe
Vitamin B6- required coenzyme
95
Why is L-lysine or L-ornithine included in the decarboxylase recipe
Amino acid substrate
96
What is the substrate in the decarboxylase test
Lysine or ornithine
97
What is the enzyme used in the decarboxylase test
Endoenzyme - decarboxylase
98
What is the product for the decarboxylase test
Cadaverine or putrescine
99
Why does glucose need to be fermented into acids first in the decarboxylase test
Need to ferment glucose to lower the pH for decarboxylase to be active
100
What are the 3 requirements for decarboxylase to be active
Low pH
Pyridoxyl is a required coenzyme
Anaerobic
101
What is a positive and negative result for the decarboxylase test
Negative
Yellow tube
Acidic pH
Bacteria metabolized sugar and produced acids which lowered the pH
Positive
Purple tube
Alkaline pH
Bacteria decarboxylated amino acid and produced diamine which raised the pH
102
What is the associated product if lysine or ornithine is used as the substrate in the decarboxylase test
lysine = cadaverine
Ornithine = putrescine
103
What is the purpose of the phenylalanine deaminase test
To determine whether bacteria can deaminate phenylalanine
104
What is the name of the media used in the phenylalanine deaminase test
Phenylalanine slant
105
What is the substrate used in the phenylalanine deaminase test
Amino acid substrate - phenylalanine
106
What is the endoenzyme used in the phenylalanine deaminase test
Phenylalanine deaminase
107
What is the product in the phenylalanine deaminase test
Ammonia + phenylpyruvic acid
108
Why can’t we use a pH indicator as a readout for the phenylalanine deaminase test
Ammonia is a base and phenylpyruvic acid is an acid
They cancel each other out
109
What is the reagent used in the phenylalanine deaminase test
Ferric chloride
Detects phenylpyruvic acid
Turns green
110
What is a positive and negative result for the phenylalanine deaminase test
Positive: turns green (phenylpyruvic acid present)
Negative: stays yellow (no phenylpyruvic acid produced)
111
If aerobic, are proteins deaminated or decarboxylated and vice versa
Aerobic: proteins are deaminated
Anaerobic: proteins are decarboxylated
112
What is the purpose of the kirby bauer method
To perform a culture and antibiotic sensitivity test on an organism
113
What makes the mueller hinton agar standardized
pH: 7.2-7.4
pH of blood
Soft agar for diffusion
4 mm in depth (lateral diffusion)
Plates incubated at 37 degrees C
Incubate for 18 hours
Amount of drug
114
How do you measure the zone of inhibition
Measure diameter of ZOI in mm
Clearing to clearing
115
What are the 3 levels of susceptibility in the kirby bauer method
Sensitive = a normal dose of an antibiotic is effective
Intermediate = a higher than normal dose of an antibiotic is required
Resistant = no acceptable dose of an antibiotic is effective
116
Are all bacteria found within the clear area around the disks dead
No they are not
They could be endospore producing and appear dormant in presence of antibiotic
117
What does it mean to be a broad spectrum drug
Antibiotic is effective to both gram + and gram - bacteria
118
What does it mean to be a narrow spectrum drug
Antibiotic is effective to just gram + not gram -
119
What organisms are most resistant to least resistant
Prions
Endospores of bacteria
Mycobacteria
Cysts of protozoa
Vegetative protozoa
Gram negative bacteria
Fungi. Including most fungal spores
Viruses without envelopes
Gram positive bacteria
Viruses with lipid envelopes
120
What is drug synergy
Two drugs display greater effectiveness together than either drug alone
121
What is the shape of staph and strep?
Cocci for both
122
What is the gram reaction of staph and strep?
gram + for both
123
What is the arrangement of staph and strep?
strap: clusters
strep: chains
124
Where is staph and strep found in the body?
staph: skin, nasal passage
strep: mouth, throat, vaginal area, GI tract
125
Is staph and strep catalase positive or negative?
staph: catalase positive (facultative anaerobe)
strep: catalase negative (aerotolerant anaerobe)
126
Reaction to cold or dryness for staph and strep?
staph: resistant
strep: sensitive
127
What is the salt tolerance for staph and strep?
staph: osmotolerant
strep: not osmotolerant
128
What is the percent of humans that carry S. epidermidis and S. aureus?
S. epidermidis: 100 percent
S. aureus: 25-30 percent of people (50 percent of healthcare workers)
129
What is the pathogenicity of S. epidermis and S. aureus?
S. epidermidis: not pathogenic
S. aureus: pathogenic
130
Is S. epidermidis and S. aureus normal flora?
S. epidermidis: normal flora
S. aureus: 2/3 of people carry it sometimes
131
Is S. epidermidis and S. aureus a fermenter of mannitol (sugar)?
S. epidermidis: no
S. aureus: yes
132
Is there a coagulase reaction with S. epidermidis or S. aureus?
S. epidermidis: negative
S. aureus: positive
133
What are the three main virulence factors of staph?
coagulase
b-hemolysin
Staphylokinase
134
What is coagulase?
forms clots (97% of staph)
- forms a clot and helps to protect bacteria from immune system
135
What is B-hemolysin?
digests red blood cells and uses the contents for food and multiplies
- digests RBCs for food
136
What is staphylokinase?
dissolves clot
- allows for spread to neighboring tissues
137
What levels of staph infection are there?
-skin diseases
- sepsis
- pneumonia
- necrotizing fasciitis
- food poisoning
138
What is the purpose of the mannitol salt agar?
to presumptively identify staphylococcus aureus and select for the growth of salt tolerant bacteria
139
Why is beef extract included in the MSA recipe?
peptones, general growth
140
Why is sodium chloride included in the MSA recipe?
selective for osmotolerant organisms
141
Why is D-mannitol included in the MSA recipe?
fermentation to produce acids, differential
142
Why is phenol red included in the MSA recipe?
pH indicator; pink = alkaline, yellow is acidic
143
What does a yellow agar on the MSA mean?
acidic pH -> bacteria metabolized the mannitol and produced acidic products
- presumptive S. aureus
- could be micrococcus (non-pathogenic)
- can determine S. aureus by coagulase
144
What does a red or pink agar on the MSA mean?
alkaline pH -> bacteria did NOT metabolize mannitol, but instead used peptones as a food source (osmotolerant)
- could be S. epidermis (-)
145
What does no growth mean on the MSA?
no osmotolerant bacteria (-)
146
What is the purpose of the coagulase test?
to test whether bacteria produce coagulase (like S. aureus)
- confirmatory test after MSA agar
147
What is the substrate for the coagulase test?
Fibrinogen
- clotting factor that is liquid and the source is rabbit plasma
148
What is the exoenzyme in the coagulase test?
Coagulase
149
What is the products for the coagulase test?
Fibrin clot (solid)
150
What is a positive and negative of the coagulase test?
positive: solid product
negative: liquid product
151
What is the purpose of the CAMP test?
to identify and confirm the presence of group B strep
- streptococcus agalactiae
152
What were the 5 organisms tested in the CAMP test?
Streptococcus agalactiae: causes serious infection in newborns
Streptococcus pyogenes: causes strep throat
Streptococcus mutans: causes cavities
Enterococcus faecalis: in GI tract
Streptococcus pneumoniae: causes pneumonia
153
What is the purpose of the bile esculin agar?
to select for the growth of enterococcus for presumptive identification
154
What are the ingredients used in the recipe for the bile esculin agar and what is each used for?
peptones; general growth
oxgall (bile salts); inhibit gram +, except enterococcus (selective)
sodium azide; inhibits gram - (selective)
esculin: has glucose and is a special food source; differential for E. faecalis
ferric citrate: turns brown with esculetin (product of esculin digestion)
155
What does growth, but not brown on the bile esculin agar mean?
growth, but not brown = enterococcus but not E. faecalis
156
What does growth and brown on the bile esculin agar mean?
presumptive E. faecalis
157
What is the M protein?
It was classified by rebecca lancefield and it has antiphagocytic properties that allow bacteria to stick to surfaces
158
What is beta hemolysis?
complete destruction of RBCs (get clear zones around colonies)
- group A and B strep
159
What is alpha hemolysis?
partial destruction of RBCs (get green zones around colonies)
- streptococcus pneumoniae
160
What is gamma hemolysis?
no damage to RBCs (no change to agar)
- S. mutans and Group D strep
161
What diseases are associated with streptococcus pyogenes and what classification of strep is it?
strep throat, scarlet fever, necrotizing faciitis
group A: beta hemolytic
162
What diseases are associated with streptococcus agalactiae and what classification of strep is it?
major neonatal pathogen, leading to pneumonia, sepsis, meningitis
group B: beta hemolytic
163
What diseases are associated with enterococcus faecalis and what classification of strep is it?
found normally in gut and infections may result in areas of the body outside of the GI tract
group D: gamma hemolytic
164
What disease are associated with streptococcus mutans and what classification of strep is it?
cavities
non-lancefield: gamma hemolytic
165
What diseases are associated with streptococcus pneumoniae and what classification of strep is it?
causes pneumonia
non-lancefield: alpha hemolytic
166
What is a positive test for the CAMP test?
look for triangular zone of enhanced beta hemolysis (synergistic)
167
What is a positive for antibiotic sensitivity in the CAMP test?
clearing around the disk
168
What is used to identify strep pneumoniae?
optochin sensitivity
169
What is used to identify strep pyogenes?
bacitracin sensitivity
170
What is used to identify strep agalactiae?
CAMP positive
171
What is used to identify enterococcus faecalis?
bile esculin positive
172
What is used to identify strep mutans?
negative to all tests
173
What is mycology?
study of fungi
174
What are the differences between yeast and mold?
yeast:
- unicellular
-colonies resemble bacteria
- reproduce by budding (asymmetrical division)
molds:
- multicellular
- colonies have velvety or fuzzy appearance
- hyphae: filaments for nutrient absorption
- filamentous - thread like
- reproduce using spores either asexually or sexually
175
What is a medically important yeast?
Candida albicans
- causes yeast infection = candidiasis
176
Who is most susceptible to candida infections?
diabetics
those with immunodeficiency
catheterized patients
infants
individuals taking antimicrobial medications
177
What is a nonpathogenic yeast?
Saccharomyces cerevisiae
- brewers yeast - used to produce beer, wine, and bread
- fermentation produces CO2 gas and ethanol
178
What are the three types of molds?
rhizopus stolonifer
- black bread mold
- hyphae are coenocytic (multinucleated cells)
- reproduce asexually (sporangiospore) or sexually (zygospore)
penicillium notatum
- medically important mold
- spores are green and hyphae are white
aspergillum niger
179
Why is fungi grown on saburaud’s dextrose agar (SAB)?
low pH of 5 or 6 encourages fungi to grow and not bacteria
180
What is the conversion for milliliters to microliters
1.0 ml = 1000 micro liters
181
What is the purpose of the plaque assay?
To determine the titer (PFU/mL) of the original phage stock solution
182
What is the clear zone in the plaque assay
Area of cell death known as a plaque because virus killed the bacteria
183
Why did we use strain B E. coli in the plaque assay
It is very sensitive to phage infection
184
What does PFU stand for?
Plaque forming unit (the number of clear zones that develop on the plate)
185
How do we know which plate to use in the plaque assay
Count the number of plaques, on a plate that has between 30-300 plaques
186
What is the equation for calculating the titer
Titer = PFU/Final Dilution Factor
115 PFU/ 10^-6 = 115 x10^6 = 1.15 x 10^8 PFU/mL
187
What are the general characteristics of helminths
Worms that are heterotrophic
Adult worms may be dioecious (male reproductive organs in one individual and female reproductive organs in another individual) or monoecious (hermaphroditic, one animal has both male AND female reproductive organs)
188
What are the two phylum for helminths
Platyheminthes (flatworms)
- Cestodes (tapeworms) flat and segmented
- Trematodes (flukes) - flat and unsegmented
Nematoda (round worms)
- Nematodes (round worms
189
What is the purpose of the hurbi biplate urinalysis
To grow bacteria from a urine sample and determine whether bacteriuria (UTI) is present and presumptively identify the cause
190
What is bacteriuria
Abnormally high levels of bacteria in the urine
191
What are the risk factors for UTI
- Catherization (put a tube in)
- female (most bacteria comes from anus, shorter route to urethra)
- sexual activity (lots of sex)
- infrequent urination (dehydration)
- age (immune system weakens as we age)
192
What causes 90% of UTIs
E. coli
193
What agar do we use for the hurbi biplate
Chromagar
Agar growth medium that turns different colors based on differences in species-specific metabolism
194
What are the selective ingredients used in the hurbi biplate
gram +
- sodium azide
- phenylethyl alcohol agar
inhibit gram -
gram -
- sodium sulfite
- oxgall
inhibit gram +
195
What are we looking for in the readout of the hurbi biplate
Look for colony type which is most prevalent and count that one type
Less than 10 = negative bacteriuria
10–100 = borderline
More than 100 = positive bacteriuria
196
How do we find the original cell density
OCD= CFU/loop volume = CFU x 1000
Ocd = 100 / 0.001 mL = 1.05 x 10^5 CFU/mL
197
What is the purpose of the transformation lab
To transform ampicillin sensitive E. coli DH5 alpha into an ampicillin resistant strain with the uptake of plasmid pBR322
198
How does DNA transformation occur
When an organism takes up foreign DNA from its surroundings
DNA is taken up through the cell wall and cell membrane
DNA must be maintained (replicated)
Organism must survive the process
199
What is a plasmid
Small, circular DNA that is not part of an organism’s chromosome (extrachromosomal)
Used to propagate foreign DNA in organisms such as E. coli
200
What are the parts of a plasmid
Origin of replication
Requirement
Selectable marker
Requirement
Multiple cloning site
Helpful but not required
201
Give a rundown of the procedure of the transformation lab
Make the bacteria competent for transformation by incubating in ice cold calcium chloride
Heat shock the cell with the plasmid DNA at 42 degrees C
Spread the E. coli on plates that contain selective growth media
LB with ampicillin
202
Predict if growth should be present on the following plates: LB + AMP: e. Coli: yes, CaCl2: yes, Plasmid: yes
Some growth (only transformed bacteria will grow)
203
Predict if growth should be present on the following plates: LB: e. Coli: yes, CaCl2: yes, Plasmid: yes
Lots of growth (ALL bacteria will grow, not just transformed bacteria)
204
Predict if growth should be present on the following plates: LB + AMP: e. Coli: yes, CaCl2: yes, Plasmid: no
No growth (no antibiotic resistance)
205
Predict if growth should be present on the following plates: LB + AMP: e. Coli: yes, CaCl2: no, Plasmid: yes
No growth or very little growth (need CaCL2 to make bacteria…)
206
What is the purpose of the differential blood cell count
To determine the percentages of the various WBCs (leukocytes) in a blood sample
207
What are the two groups of leukocytes
Granulocytes (which have prominent cytoplasmic granules)
Neutrophils
Eosinophils
Basophils
Phil = filled with granules
208
Agranulocytes (which lack granules)
Lymphocyte
Monocyte
209
What is the general relative abundance of white blood cells
Never Let Monkeys Eat Bananas
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
210
Neutrophil: What is the abundance in blood (%)
55-65
211
Neutrophil: What is the diameter
12-15 micro meters
212
Neutrophil: Nucleus
2-5 lobes
213
Neutrophil: cytoplasmic granules
Present but stain poorly; contains antimicrobial chemicals
214
Neutrophil: functions
Phagocytosis and digestion of (usually) bacteria
215
Eosinophil: what is the abundance in blood percentage
1-3
216
Eosinophil: What is the diameter
12-15 micro meters
217
Eosinophil: What is the nucleus
2 lobes
218
Eosinophil: cytoplasmic granules
Present and stain red; contains antimicrobial chemicals and histamine
219
Eosinophil: function
Phagocytic, fight against worm infections, some involvement in allergy response
220
Basophil: abundance in blood percentage
0.5 -1
221
Basophil: diameter
12-15 micrometers
222
Basophil: nucleus
Unlobed or 2 lobes
223
Basophil: cytoplasmic granules
Present and stain dark purple; contains histamine and other chemicals
224
Basophil: functions
Release histamine during allergic response
225
Lymphocyte: abundance in blood
25-33
226
Lymphocyte: diameter
7-18 micrometers
227
Lymphocyte: nucleus
Spherical (little cytoplasm)
228
Lymphocyte: cytoplasmic granules
Absent
229
Lymphocyte: functions
Specific acquired immunity
B cells: produce antibodies
T cells: direct immune response
230
Monocyte: abundance in blood percentage
3-7
231
Monocyte: diameter
12-20 micrometers
232
Monocyte: nucleus
Horseshoe (cytoplasm is prominent)
233
Monocyte: cytoplasmic granules
Absent
234
Monocyte: functions
Phagocytosis
Macrophages
235
What is the purpose of blood typing
To determine the blood type of a person
236
What is an antigen
Any substance that can elicit an immune response
237
What do you need to have an O blood type
Two recessive alleles
238
What antibodies are produced in blood type O
Anti A and Anti B
239
What antibodies are produced in blood type A
Anti B
240
What antibodies are produced in blood type B
Anti A
241
What is the universal donor and universal recipient
Donor: O
Recipient: AB
242
What is hemagglutination
A general term applied to any agglutination test in which the clumping of red blood cells indicates a positive reaction
243
What does agglutionation indicate
Type A blood of donor + anti A antibody of type B recipient = binding
Clumping indicates A antigens are present
Unsuccessful blood typing
244
What does Rh+ indicate
Has clumping factor
Does not produce anti-Rh antibodies
Can receive rh+ or rh- blood
Genotype can be +/- or +/+
245
What does Rh- indicate
Lacks clumping factor
Produces anti-Rh antibodies
Can receive Rh- blood ONLY
Genotype is -/-
246
Enterobius vermicularis: Disease
Enterobiasis (pinworm) - itchy rectum
247
Enterobius vermicularis: Cestode, Trematode, Nematode
Nematode
248
Enterobius vermicularis: Monoecious or dioecious
Dioecious
249
Enterobius vermicularis: Stages to identify on exam
Female adult
Eggs
250
Enterobius vermicularis: Infectious stages and Mode of Transmission
Eggs; swallow or inhalation of eggs through contaminated clothing, bedding, or fingers
251
Enterobius vermicularis: Approximate size of the adult worm
about size of grain of rice 1cm
252
Enterobius vermicularis: Symptoms
itchy rectum
many people asymptomatic
253
Enterobius vermicularis: Treatment
albendazole
mebandazole
(2 doses, 2 weeks apart)
254
Enterobius vermicularis: Diagnosis and Additional Notes
cellophone tape test looking for eggs from the perianal region under the microscope
255
Ascaris lumbricoides: Disease
Ascariasis + Ascariasis pneumonia
256
Ascaris lumbricoides: Cestode, Trematode, Nematode
nematode
257
Ascaris lumbricoides: Monoecious or dioecious
dioecious
258
Ascaris lumbricoides: Stages to identify on exam
male and female adult
eggs
259
Ascaris lumbricoides: Infectious stages and Mode of Transmission
Ingestion of fertilized eggs in fecally contaminated uncooked food or water
260
Ascaris lumbricoides: Approximate size of the adult worm
males: 31 cm
females: 35 cm
261
Ascaris lumbricoides: Symptoms
usually asymptomatic OR
abdominal discomfort, heavy infection can cause intestinal blockage, or cough
262
Ascaris lumbricoides: Treatment
albendazole
mebandazole
(1-3 days)
263
Ascaris lumbricoides: Diagnosis and Additional Notes
identifying eggs in stool sample
264
Necator americanus: Disease
Hookworm -> infection leads to anemia
265
Necator americanus: Cestode, Trematode, Nematode
Nematode
266
Necator americanus: Monoecious or dioecious
Dioecious
267
Necator americanus: Stages to identify on exam
adult
filariform larvae
eggs
268
Necator americanus: Infectious stages and Mode of Transmission
filariform larvae; walking barefoot on contaminated soil and filariform larva penetrate skin
269
Necator americanus: Approximate size of the adult worm
males: 0.5-0.9 cm
females: 1 cm
270
Necator americanus: Symptoms
itching and localized rash, may be asymptomatic, OR abdominal pain, diarrhea, loss of appetite, weight loss
271
Necator americanus: Treatment
albendazole
mebendazole
(1-3 days)
272
Necator americanus: Diagnosis and Additional Notes
larva penetrate skin -> blood bessels > heart > lungs > swallowed and adults develop in intestine; identifying eggs in stool sample
273
Schistosoma mansoni: Disease
Schistomiasis
274
Schistosoma mansoni: Cestode, Trematode, Nematode
Trematode
275
Schistosoma mansoni: Monoecious or dioecious
Dioecious
276
Schistosoma mansoni: Stages to identify on exam
males and female adult
cercaria
eggs
277
Schistosoma mansoni: Infectious stages and Mode of Transmission
cercaria can penetrate skin of persons who came in contact with contaminated water when wading, swimming, or bathing
278
Schistosoma mansoni: Approximate size of the adult worm
1.5 - 2cm
279
Schistosoma mansoni: Symptoms
fatigue and anemia
most are asymptomatic, itching and rash, fever, chills, cough, and muscle aches; chronic ab pain, enlarged lungs, blood in stool or urine, or problem passing urine
280
Schistosoma mansoni: Treatment
praziquantel for 1-2 days
281
Schistosoma mansoni: Diagnosis and Additional Notes
identifying eggs in stool or urine sample under the microscope
282
Taenia solium: Disease
Taeniasis + Cysticerosis
283
Taenia solium: Cestode, Trematode, Nematode
Cestode
284
Taenia solium: Monoecious or dioecious
Monoecious
285
Taenia solium: Stages to identify on exam
adult scolex
adult gravid proglottid
cystericus
eggs
286
Taenia solium: Infectious stages and Mode of Transmission
Taeniasis: ingestion of raw or uncooked meat containing the infective cystericerol
Cysticercosis: ingestion of egg of T. solium, get cysticercosis of brain, eyes, and muscles
287
Taenia solium: Approximate size of the adult worm
up to 5 meters
or up to 25 m
288
Taenia solium: Symptoms
taeniasis: many asymptomatic but can cause ab pain, loss of appetite, weight loss, and upset stomach
cysticercosis: seizures and muscles or eye damage
289
Taenia solium: Treatment
praziquantel or niclosamide
290
Taenia solium: Diagnosis and Additional Notes
identifying eggs or proglotting in stool sample
