Preparation of Culture Media, Inoculation & Incubation Techniques Flashcards
1
Q
Increase in cell number through binary fission
A
Microbial growth
2
Q
cell simply splits into two cells, producing two identical cells
A
Binary fission
3
Q
Difference of binary fission from mitosis
A
No mitotic spindle form
4
Q
the average time between two consecutive generations in the lineages of a population
A
generation time
5
Q
generation time E.coli
A
30 to 180 mins
6
Q
generation time M. tuberculosis
A
12 hours
7
Q
T. pallidum
A
33 hours in rabbit testes
8
Q
M. leprae
A
10 days in armadillos
9
Q
Cocci synthesize new cell walls
A
septal region
10
Q
Synthesize new cell wall entire surface of the cell
A
Rod shape bacteria
11
Q
Logarithmic representation of bacterial growth
A
2^n
12
Q
cell form an initial outgrowth that enlarges until its size approaches that of a parent cell and then it separates
A
Budding
13
Q
the initial outgrowth
A
a bud
14
Q
microbes that grow and multiply in or on a culture medium
A
culture
15
Q
A group of descendants of an original cell
A
colony
16
Q
contains a single microbial species
A
pure culture
17
Q
other name of pure culture
A
axenic culture
18
Q
how the surface of the colony appears
A
texture
19
Q
Texture types
A
smooth
glistening
mucoid
slimy
dry
powdery
flaky
20
Q
Transparency
A
Transparent
Translucent
Opaque
21
Q
You can see through them
A
Transparent
22
Q
Light pass through them
A
Translucent
23
Q
solid-appearing
A
opaque
24
Q
Intracellular pigments
A
color/pigmentation
25
Many bacteria do not produce any pigment and appear __________
white or gray
26
Magnifying tool
Colony counter
Dissecting microscope
27
Colony size
Punctiform <1 mm
Small 1-2 mm
Medium 3-4 mm
Large >5 mm
28
Colony forms
Circular
Rhizoid
Irregular
Filamentous
29
Colony Margin
Entire
Undulate
Lobate
Filamentous
Curled
30
Colony elevation
Flat
Raised
Convex
Pulvinate
Umbonate
31
Rely on organic compounds as source of food to provide energy and carbon
Heterotrophs
32
Most lab bacterias we work with
Mesosphile
33
Mesophile/Mesophilic temperature
20 to 40 degrees celsius
34
Body Temperature
37
35
Room temperature
25
36
The lowest temperature at which an organism is able to conduct metabolism
Minimum growth temperature
37
Temperature at which an organism's metabolic activities produce the highest growth rate
optimum growth temperature
38
The highest temperature at which an organism continues to metabolize
maximum growth temperature
39
when temperature exceed maximum growth temperature, the organism's _________
protein are permanently denatured and dies
40
P S Y C H R O P H I L E S other name
cryophiles
41
sensitive to temperatures over 20 ̊C
optimum growth at
15 ̊C or below
P S Y C H R O P H I L E S
42
− optimum growth at 20 ̊C to 30 ̊C
P S Y C H R O T R O P H S
43
− optimum growth at 25 ̊C to 40 ̊C
M E S O P H I L E S
44
− optimum growth at 50 ̊C to 60 ̊C
T H E R M O P H I L E S
45
P S Y C H R O T R O P H S Other name
Cold-tolerant bacteria or archaea
46
grow at 80 ̊C or higher
Extreme Thermophiles
47
M E S O P H I L E S other name
moderate temperature-loving
48
T H E R M O P H I L E S other name
Heat-loving
49
T H E R M O P H I L E S example
Bacillus stearothermophilus
50
Extreme Thermophiles Example
Geogemma barosii
51
Geogemma barosii
85 to 121
2 hours in 130
52
Can destroy most microbes
> 60 degrees
53
slow microbial growth
50 to 60
54
rapid microbial growth; produce toxins
25 to 50
55
grow at very low pH (0.1 to 5.4)
A C I D O P H I L E S
56
Organs with low pH
Vagina and stomach
57
causative agent of peptic ulcer
Helicobacter pylori
58
A C I D O P H I L E S example
Lactobacillus acidophilus
59
− grow at pH 5.4 to 8.5
N E U T R O P H I L E S
60
Lactobacillus produce
lactic acid
61
− grow at high pH (7 to 12)
A L K A L I P H I L E S
62
Most bacteria, protozoa, and pathogens grow best at what pH
Neutral
63
A L K A L I P H I L E S example
− Vibrio cholerae
cholera
− Alkaligenes faecalis
− Agrobacterium spp.
64
Pressure difference needed to stop the flow of solvent across a semipermeable membrane
Osmotic pressure
65
require very high salt conc. 20 to 30%
O B L I G A T E
H A L O P H I L E S
66
O B L I G A T E
H A L O P H I L E S
other name
extreme halophiles
strict halophiles
67
Can survive month in dry conditions due to the ability of its cell wall to retain water
Mycobacterium tuberculosis
68
− do not require high salt conc. for growth
− tolerates 2-15% salt concentration
F A C U L T A T I V E
H A L O P H I L E S
69
Salt-water principle
Water follows salt
Sodium attracts water
70
No net movement of water
Cells in isotonic solution
71
Shrink because of the net movement of water out of the cell
Cell in hypertonic solution
72
Undergo gain of water
Cells in hypotonic solution
73
Solute concentration inside and outside are the same
Isotonic solution
74
Have higher solute concentration than inside the cell
Hypertonic solution
75
Has a lower solute concentration
Hypotonic solution
76
Hypo comes from
Latin word under or low
77
− organisms which derive energy from organic carbon sources
− may need to consume other living organisms to
live
C H E M O H E T E R O T R O P H S
78
Organisms that obtain energy by the oxidation of electron donors in their environment
Chemotrophs
79
Organism that cannot manufacture its own food by carbon fixation and therefore derives its intake from other sources of organic carbon, mainly plant or animal matter
Heterotroph
80
− organisms which use inorganic energy sources to synthesize organic compounds
− primary carbon source is carbon dioxide
C H E M O A U T O T R O P H S
81
organism that can produce its own food using light, water, carbon dioxide, other chemicals
Autotroph
82
makes up 14% of
dry cell weight
N I T R O G E N
83
Nitrogen is to build __________
Proteins and nucleotides
84
N I T R O G E N sources
proteins,
ammonium,
nitrogen gas,
nitrates
85
Some bacteria can reduce nitrogen gas to
ammonia
86
− used to form proteins and some vitamins
S U L F U R
87
S U L F U R sources
proteins,
hydrogen sulfide,
sulfates
88
Sulfur is used to synthesize sulfur-containing amino acids and vitamins like
Thiamine - Vitamin B1
Biotin - Vitamin H
89
used to form DNA, RNA, ATP, and phospholipids
P H O S P H O R U S
90
P H O S P H O R U S sources
Inorganic
phosphate salts,
Buffers (from culture medium)
91
phospholipids are found in _______
cell membranes of bacteria
92
Growth requirement for aerobic microorganisms
O X Y G E N
93
O X Y G E N sources
Atmosphere,
Water
94
Require oxygen to live
O B L I G A T E
A E R O B E S
95
O B L I G A T E
A E R O B E S
example
Mycobacterium tuberculosis
96
Can use oxygen, but can grow in its absence
F A C U L T A T I V E
A N A E R O B E S
97
F A C U L T A T I V E
A N A E R O B E S
example
Escherichia coli
98
F A C U L T A T I V E
A N A E R O B E S
has ________ enzymes that allow toxic forms of oxygen to be neutralized
catalase
SOD (superooxide dismutase)
99
− Can’t use oxygen and are harmed by the
presence of toxic forms of oxygen
O B L I G A T E
A N A E R O B E S
100
O B L I G A T E
A N A E R O B E S
example
Clostridium tetani
101
Can’t use oxygen, but can grow in its absence
Tolerates presence of oxygen
A E R O T O L E R A N T
A N A E R O B E S
102
A E R O T O L E R A N T
A N A E R O B E S
example
Streptococcus pneumoniae
103
− Require oxygen at low concentrations
M I C R O -
A E R O P H I L E S
104
A E R O T O L E R A N T
A N A E R O B E S
also have _________
SOD
105
M I C R O -
A E R O P H I L E S
example
Helicobacter pylori
Campylobacter jejuni
106
Toxic forms of oxygen
Hydrogen peroxide
Superoxide radicals
107
converts superoxide to oxygen & hydrogen peroxide
Superoxide dismutase:
108
converts hydrogen peroxide to water & oxygen
Catalase:
109
breaks down hydrogen peroxide
Peroxidase:
110
− CO2-loving
− require increased concentration of CO2 and lower levels of O2
C A P N O P H I L E S
111
C A P N O P H I L E S
Example
Neisseria gonorrhoeae
Haemophilus influenzae
112
Growing capnophiles
Candle jar
CO2 generating packet
113
Trace elements
− K
− Mg
− Ca
− Fe
− Cu
− Mo
− Zn
114
Used for cell wall synthesis of gram positive bacteria
Calcium
115
BACTERIAL GROWTH CURVE
1. Lag phase
2. Log phase
3. Stationary phase
4. Death or decline phase
116
No increase in number of living bacterial cells
(Phase of intense metabolic activity)
1. Lag phase
(1 hour to several days)
117
Exponential increase in number of living bacterial cells
(Period of most rapid growth)
2. Log phase
118
plateau in number of living bacterial cells; rate of cell division and death roughly equal
3. Stationary phase
119
exponential decrease in number of living bacterial cells
4. Death or decline phase
120
CULTURE MEDIA Characteristics:
i. Contains appropriate nutrients
ii. Sterile
iii. Incubated at appropriate temperature
121
act of introducing microorganism or suspension of microorganisms into a culture medium
Inoculation
122
substance containing nutrients in which cells or microorganisms can be grown; may be solid, liquid or gel
CULTURE MEDIA
123
microbial cells added to a medium in order to start a culture
Inoculum:
124
nutrient material that contains a solidifying agent
SOLID MEDIA
125
− Allows for isolation and examination of colonies
− Difficult to transport and store
Agar Plate
126
Free from any microorganism
Sterile
127
− Inoculating needle is used to “stab” the solid medium
− For microorganisms that prefer reduced O2
Agar Deep
128
− Tube of solid medium at an angle for higher surface area
− For easy storage and transportation
− Use for long term maintenance of isolates
Agar Slant
129
CULTURE MEDIA common ingredient
Peptone
Beef extract
Yeast extract
Distilled water
agar
130
− Melts at above 95 ̊C; once melted, does not solidify until it reaches 40 ̊C
− Cannot be degraded by most bacteria
A G A R
131
The manner in which the inoculum is spread out into the cultured medium
Streaking
132
A G A R is the ____________ obtained from the cell walls of some species of red algae
Unbranched polysaccharide
133
some species of red algae
i. Gracilaria compressa
ii. Gelidium cartilagineum
iii. Gelidium amansii
134
nutrient material that lacks a solidifying agent
LIQUID MEDIA
135
− Liquid medium in a tube, bottle,
or flask
− Applicable if high concentration
of bacteria is desired
Broth
136
Who first used agar
Robert Koch
137
nutrient material that contains <1% solidifying agent
SEMISOLID MEDIA:
138
Typical agar concentration
1.5 to 2 %
139
the concentration of solid media
3%
140
Agar solidifies at what temperature
37
141
Another solidifying agent
Gellan gum
142
Solid media function
1. Used to grow microorganism in their full physical form
2. Used to prepare bacterial pure culture
3. Use to isolate bacteria to study colony characteristic
143
Inoculating techniques
Stabbing
Stab and streak
Streaking
144
Irish moss (Red Algae)
Chondrus crispus
145
may be done in media with dilute agar concentration
motility test
146
useful in demonstrating motility of bacteria
SEMISOLID MEDIA:
147
For general purposes of
preservation, isolation, storage of
cultures and antibiotic sensitivity
tests
ALL - P U R P O S E
M E D I A
148
ALL - P U R P O S E
M E D I A
examples
− Peptone water
− Nutrient broth
− Nutrient agar
− Bismuth Sulfite Agar
149
Allows growth of desired
microorganism while suppressing
the growth of unwanted
microorganisms
S E L E C T I V E
M E D I A
150
S E L E C T I V E
M E D I A
example
− Saboraud’s dextrose agar
− Brilliant green agar
− Thayer-Martin agar
− Lowenstein Jensen Media
151
pH 5.6 discourages bacterial growth
for fungi
Saboraud’s dextrose agar
152
inhibits G+ bacteria and allows for the growth of G- salmonella
Brilliant green agar
153
for the growth of Neisseria gonorrheae
Thayer-Martin agar
154
Used to isolate Salmonella typhi
Bismuth Sulfite Agar
155
Used to isolate Mycobacterium tuberculosis
Lowenstein Jensen Media
156
Used for differentiating between
bacteria by using an identification
marker for a specific type of
microorganism
D I F F E R E N T I A L
M E D I A
157
D I F F E R E N T I A L
M E D I A
examples
− Mannitol salt agar
− MacConkey agar
− Blood agar
158
Mannitol salt agar
Staphyloccocus aureus -> halophile
159
MacConkey agar
Salmonella typhi
160
Uses hemolytic reaction
Used to distinguish bacteria that destroy red blood cells
Blood agar
161
Blood agar usually contains
sheep or horse blood
162
contains 7.5 % sodium chloride
Both selective and differential
has a pH indicator
Mannitol salt agar
163
Mannitol fermentation
pink to yellow positive
164
consider as complex, selective, and differential media
contains bile salt, and crystal violet that inhibits growth of g+ bacteria
use lactose
MacConkey Agar
165
The growth in _______ indicates _______ , while _______ does not
pink; lactose utilization; yellow
166
Lactose fermenters examples
Enterobacter spp
Citrobacter spp
Klebsiella spp
167
Lactose fermenters will produce
acid
168
test the ability of an organism to produce hemolysin, enzyme that damage/lyse red blood cells
Blood agar media
169
Blood agar media members
Staphylococcus
Streptococcus
Enterococcus
170
Partial hemolysis
Alpha hemolysis
171
Complete hemolysis
Beta hemolysis
172
No hemolysis
Gamma hemolysis
173
the destruction of red blood cells
hemolysis
174
Alpha hemolysis examples
Streptococcus pneumoniae
Streptococcus mutans
175
Beta hemolysis examples
Staphylococcus aureus
Streptococcus pyogenes
Listeria ivanovii
176
Gamma hemolysis examples
Escherichia coli
Klebsiella pneumoniae
Enterococcus faecalis
177
Favors growth of a particular
microorganism over others by
providing essential nutrients
E N R I C H M E N T
M E D I A
178
E N R I C H M E N T
M E D I A
examples
− Chocolate agar
− Loeffler’s medium
− Brain heart infusion agar
179
complex or particular nutrient requirement
fastidious organism
180
Chocolate agar example
Haemophilus influezae
181
Loeffler’s medium example
Corynebacterium diphtheriae (Corynebacteria)
182
Brain heart infusion agar examples
Streptococci
Pneumococci
Meningococci
183
Made up of lysed RBCs
Chocolate agar
184
Haemophilus influenzae
Pneumonia and sepsis
185
Prepared from pure chemical
substances of known composition
and concentration
C H E M I C A L L Y
D E F I N E D
M E D I A
186
C H E M I C A L L Y
D E F I N E D
M E D I A
example
− Peptone water
− Modified Korthof’s medium
− Firm agar
187
Exact components are difficult to
estimate because of complex
ingredients
C O M P L E X
M E D I A
188
C O M P L E X
M E D I A
example
− Nutrient agar
− Trypticase soy agar
− Eosin methylene blue agar
189
use to culture most chemoheterotrophic microorganism
C O M P L E X
M E D I A
190
Eosin methylene blue agar is used for
Isolation of the fecal coliforms (E.coli)
191
Eosin methylene blue agar contains lactose and dyes _______ and _______
Eosin and methylene blue
192
Microorganism that cannot grow in culture media
Obligate parasites
193
SPECIAL CULTURE TECHNIQUES armadillos
Mycobacterium leprae (leprosy)
194
SPECIAL CULTURE TECHNIQUES rabbit testicles
Treponema pallidum (syphilis)
195
SPECIAL CULTURE TECHNIQUES host cells only
Obligate parasites
STD causing
chlamydia
196
SPECIAL CULTURE TECHNIQUES hen's egg
Influenza virus
197
SPECIAL CULTURE TECHNIQUES Cell culture
Mycoplasma spp
198
A method of isolating a culture by spreading microorganisms over the surface of
a solid culture medium.
S T R E A K
P L A T E
M E T H O D
199
A method of inoculating a solid nutrient medium by mixing bacteria in the melted
medium and pouring the medium into a Petri dish to solidify.
P O U R
P L A T E
M E T H O D
200
A plate count method in which inoculum is spread over the surface of a solid
culture medium.
S P R E A D
P L A T E
M E T H O D
