mbio exam 2 Flashcards
(188 cards)
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must Have O2 to survive
Obligate aerobes
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can grow in the presence or absence of O2
Facultative anarobes
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Do not need O2 but can survive in presece of O2
Aerotolerant & anarobes
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Survive only when levels of O2 are low
microaerophiles
46
Survive only when levels of CO2 are high
Caphophiles
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Any o2 eill kill the bacteria
Obligate Anarobes
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the ability or inablity to live in the presence of O2
aerotolerance
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Bacteria will grow in the tube where the O2 level is _ for that organism.
optimal
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Autoclaving a capped tube of media will remove most of the free O2 . as the media cools O2 will start to _ back into the media. Creating a __ where there will be __ O2 at the surface of the media while the bottem of the tube will have ___ O2.
Diffuse
Concentration Gradiant
Higher
No
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Adjusting to new enviroment ___ metabolic activity ( no division) and length of lag depends on how extream of an enviroment change.
This is the-Lag Phase
High
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Bacteria begin to divide at maximum growth rate the growthis __ and will last as long as sufficent __ and favorable conditions are present
this is the- Log Phase
exponitial
nurients
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Eventually nutrients become limited and ___ products begin to accumulate. In this phase the ___ rate equals the ___ .
this is the- Stationary phase
Wast
growth
death rate
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All ___ have been depleted and the accumulated wast products make the enviroment \_\_\_.
this is the -Death Phase
Nutrients
Toxic
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Phases of the bacterial growth curve
lag phase
log phase
Stationary phase
Death phase
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the bacterial growth curve provides ______ information about a bacterial species.
length fo time in each phase of growth
optimal growth conditions
mean generation time of the organism.
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bacterial are ___ and can be found in many diffrent habitates.
Ubiquitous
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Some Possible conditions that bacteria may encounter \_\_\_\_\_,\_\_\_\_, and \_\_\_\_\_
temps
PH
Osmotic pressure
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Minimal, optimal and maximal temperatures at which bacterium can grow
Cardial Temperatures
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Grow @ range of \< 20 C
found in ___ , and \_\_\_\_.
Human Pathogen?
Psychrophiles
23-86 F
alpine soil
Ice feilds
No
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Grow at range of 0 C- 30 C \_\_\_\_F. found in \_\_\_\_, ____ and \_\_\_\_.
Human Pathogens?
Psychrophiles
23-86 f
soil, surface water, and food.
Yes can be human pathogens
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Grow at range of ~15 c to~40 c \_\_\_\_\_F
Human Pathogens?
Mesophiles
59-130 F
Yes can be human pathogens and cause deisease.
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Range ~40 to ~75 C \_\_\_\_\_\_F found\_\_\_\_.
Thermophiles
104-167 F
found in hot springs
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Grow at a range of ~65C to ~110 C \_\_\_\_\_F. where are they found?
Hyperthermophiles
149-230 F
ocean floor
thermal vents and ridges
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PH is the ______ of ______ ions measured on the _______ scale?
Concentration
H+
logmarihmic
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As H+ increases the PH ______ and the solution becomes \_\_\_\_\_\_?
goes down
acidic
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As H+ decreases the PH ______ and the solution becomes \_\_\_\_\_\_?
goes up
Basic
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three major catogories for bacteria based on PH?
Acidophile
Nuetrophile
Alkaphile
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grow well in acidic enviroments...
Ph of \<= 4.5-5.5
Acidophile
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Stomach acid has a PH Of?
1-2=PH
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Grow well at neutral Ph between 5.5 & 8
Nuetrophile
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Most human pathogens are \_\_\_\_\_\_
Neutrophiles.
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Human blood has a PH of ?
PH- 7.4
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Grow well in basic/ a;la;ome PH of \>= 8-8.5
Alkaphile
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H2o is essential for all forms of life including bacteri. Bacteria need water to maintain\_\_\_\_\_\_\_. this is necessary so that cells do not _____ or \_\_\_\_\_\_.
Turgor pressure
lysis
plasmolysis
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To regulate this turgor pressure bacteria use _____ and _____ ions to maintain a _______ in which there are more ions inside the cell than outside.
Na+
K+
Concentration gradiunt
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We know that through the process of _____ water moves from areas of ____ ion concentration to areas of \_\_\_\_- concentration to maintain/ achieve equilibrium.
Osmosis
low
high
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\_\_\_\_\_\_ is the mesure of the force with which H2o flows during osmosis.
Osmotic Pressure.
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three types of Osmotic enviroments a bacteria can encounter.
Hypotonic
Isotonic
Hypertonic
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Higher solute inside the cell than out
low osmotic pressure
\_h20 moves into the cell
cell wall lysis
Hypotonic
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equal solute can go inside and outside the cell
Isotonic
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Low solute can be inside than outside the cell
High osmotic pressure
H20 moves out of cell
cell wall plasmolysis
Hypertonic
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Salinity amount of salt in a solution can affect the growth of bacteria. most bacteria grow in concentration less than ____ %
3%
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Human pathogens prefer the \_\_\_\_\_% of ______ and \_\_\_\_\_.
.9% nacl
blood
Urine
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certain bacteria love and grow optimally in Nacl concentrations greater than 3% and are called \_\_\_\_\_\_\_\_.
Haleophiles.
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In addition you also find Extream Halophiles that only survive in Nacl concentration of ____ to \_\_\_\_.
15
25
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If bacteria can grow over a wide rage of Nacl cnacentrations it is called \_\_\_\_\_\_\_\_\_.
osmotolerant bacteria.
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Direct count metohods
Plate count
Filtration
Most probable Numbers
Direct Counts
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indirect count metods
Metabolic activity
dry weight measures
Turbidity
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a Spectrophotometer is used to measure __________ of a sample.. A _________ of light is passed through the sample where the photo detector converts the light energy into an ________ which can be used to mesure the amount of light ________ by the sample.
turbidity
Monochromatic ( single wavelenght)
electric signal
transmittence % of absorbance
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\_\_\_\_\_\_ or _______ is the amount of light neither transmitted or reflected by the sample.
absorbance
optical density
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absorbance range from 0-1.99 A anything above that needs to be \_\_\_\_\_\_.
diluted
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There is a ______ relationship between the amount of light absorbed and the concentration of cells in a sample.
Direct -linerar
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Low absorbance =
low # of bacteria
Lowest turbidity
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high absorbance
High # of bacteria
Higher turbidity
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what do Mc farland standards repersent?
A set of refrence smaples of diffrent turbidity that can be used to estimate the # of bacterial cells in a sample.
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Bacteria replicate by
Bianery fission
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is the time it takes bacteria to get from 2^x to 2^x+1
doubling /generation time
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it takes _____ salmonella to cause illness and theier doubling time is about _____ min.
500
20
100
NF=Ni(2^n)
Nf=#of cells at final time point
Ni= # of cells at initial time point
N=#of generations
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doubling times vary by species and enviroment factors and can range from ______ min to ______ days . the average doubling time is _____ to ____ min.
10 min
30 days
20 min
60 min
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\_\_\_\_\_\_\_ can be used to check the various effects of enviromental changes to a bacteriums growth.
closed growth system
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In a closed growth System No ______ are added and No ________ are removed
nutrients
waste products
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Mesaures a bacterial population
Mesaures the number of viable cells (cfuS)
Plate counts
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Stattistical estimates metod for mesuring a population ( mpn /100 ml)
Most portable Numbers
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Mesaures a bacteria population based on the fact that bacterium cannot pass through pors of membran filter.
filtration
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Mesaure the number of bacteria cells in a sample.
manual counts using Petroff Hausser counting chamber
Direct Counts
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estimates bacterial #'s by measuring the metabolic activity of a bacterial population... production of O2 and acide.
Metabolic activity
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Estimate bacterial #'s by mesuring the weight of bacterial population
dry weight mesure
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Estimate bacterial #'s by mesureing turbidity of growing bacteria.
turbidity.
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what are the three fluid thioglycollate medium
Sodium Thioglycollate
L-cystine
Resazurin
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reducing agent for Sodium Thioglycollate
reduces O2----\>H2O
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reducing agent for L-cystine
reduces O2----\> H2O
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reducing agent for resazurin
oxidation reduction indicator
red/pink= presence of O2
yellow/straw= absence of O2
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can aerotolerance be determined with agar plates? how?
yes however plates have to be incubated under arobic and anarobic conditions.
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Methylene Blue indicator strip in gas jar turns\_\_\_\_\_ in the ____ of \_\_\_\_\_.
Blue
presence
O2
white = absence of O2
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In the Anarobic Jar experiment if conditions are not achived what happens?
strip remains blue - means there is a hole in the bag
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what does the Oxidation Fermintation test tell Us?
which catabolic pathway a bacteria uses
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O2 is the final electron acceptor
Must be in an AEROBIC enviroment- oxygen
Aerobic respiration/Oxidation
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Other ORGANIC molecule is the final electron acceptor
either AEROBIC or ANAEROBIC
(presence) or ( absence)
Fermentation
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occures only in the presence of O2
Aerobic respiration
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can occure in the presence or absence of O2
Fermentation.
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produces small acidic intermediat compounds
Arobic respiration.
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large acid compounds produced
Fermentation
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Media has two important Componets
Carbohydrates- glucose, lactose, sucrose ect..
Ph indicator- bromothmol Blue
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Oxidation Media is \_\_\_\_\_\_... with a PH of\_\_\_\_
Green
PH 7.1
oil layer is added to make enviroment anaerobic and promote fermentation.
when ph drops below 6 = more acidic.... the media turns yellow.
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Oxidation results ( only pathway used)
Oil
Green
no breakdown
no acid= PH
glucose cannot be broken down under anarobic conditions.
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Oxidation test possible outcomes
No Oil
Yellow @ Top ( O2 present)
Some breakdown
Some Acid (lower PH)
glucose broke down ----\> moderate amounts of acidic compounds made =( drop in PH)
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Fermentation Results
Oil
Yellow
Breakdown
Acid ( lower PH)
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Fermentation Test results
No Oil
Yellow
breakdown
Acid Lower PH
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No utilization of Carbohydrate test results
Oil
green
no breakdown
No Acid= PH
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No utilization of carbohydrate
No Oil
Green
no breakdown
No Acid=PH
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No utilization of Carbohydrate
or
Blue- green
indicating breakdown of protiens
basic/ alkaline Higher PH
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what does the phenol Red Broth tell us?
it determins what carbohydrates a bacterium can ferment.
this is a differental test since the ability of one species to ferment a carbohydrate may be diffrent than that of another.
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\_\_\_\_\_\_\_ is not the only sugar that can be used to start glycolysis.
Glucose
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the end products for fermentation are usally \_\_\_\_\_\_\_\_\_.
acidic compunds
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\_\_\_\_\_\_ can also be an end product of fermentation
gas
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three componets of Phenol red broth
Base- no carbohydrate in the media ( so you can add the one you want to test.
PH indicator- phenol Red
Durham Tube- to check for gas production- places upside down to see if gas bubbles are produced
139
Four possible outcomes for Phenol red Broth
Fermentation - no gas
fermentation - gas produced
no fermentation -no gas
no fermentation - protien breakdown
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test results for fermentation - no Gas
yellow
lower in PH acid
No Gas
Record A/-
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test results fermentation Gas produced
yellow
Acid ( ower in PH)
Gas
Record- A/ G
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Fermentation test results
No fermentation - No Gas
Red
No Acid = PH
Record -/-
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fermentation test results
no fermentation - protien breakdown
bright pink
Basic/alkaline ( higher PH)
record K
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Catalase Test tells us what?
determins the presence or absence of the enzyme catalase in the bacterium
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what is the diffrence in Aerobic and Anaerobic Respiration?
the diffrence is the final electron acceptor.
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the final electron acceptor in Aerobic respiration is ?
O2
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the final electron acceptor in Anaerobic Respiration
Nonoxygen molecule
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electrons are passed down a chain of molecules that alternate between becoming _______ or \_\_\_\_\_\_\_\_.
oxidized
reduced
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During this passing of electrons some can escape called\_\_\_\_\_\_\_\_. when this occurs a ____________ will be produced.
premature electron leakage
superoxide O2
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\_\_\_\_\_ is extreamly toxic to bacteria and they must be able to _____ to survive.
O2
break it down
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Bateria use a enzyme called\_\_\_\_\_\_\_\_\_\_ the reaction is?
SuperOxide Dismutase
O2 + SOD ---\> O2 + H2O2
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\_\_\_\_\_ is not quite as toxic as \_\_\_\_- but bacteria must still break it down further. the reaction is?
H2O2
O2
H2O2 + Catalase ---\> H2O2 + O2 ( gas)
153
catalse test has two important components
bacteria - smered on glass slide
H2O2- test reagent
two possible outcomes = catalase positive
catalase negative
154
results for catalase negative
no catalase enzyme
No O2 gas produced
155
results for catales test
catalase positive
catalase enzyme present
O2 gas produced
156
what does the oxidase test tell us?
the oxidase test determins the presence or Absence of cytochrome C oxidase in the ETC of the bacterium.
157
Etc's can very from organisum to organisum. some are similar to those found in eukaryotic mitochondria. These mitochondria like ETC's consist of 4 enzyme complexes used to transfer electrons. The last complex is Complex IV ( cytochrome C Oxidase) and specifically transfers from cytochrome C to O2.
oxidize - loss of electron
reduction - gain of an electron
158
\_\_\_\_\_Catalyzes movement of an electron from one molecule to another?
Cytochrome C Oxidase
159
\_\_\_\_\_\_ is a chemical that donats an electron and when doing so changes color. In our experiment it was Tetramethyl-p-phenylenediamine (TMPD)
Chromogenic Reducing Agent
160
reaction of oxidase test
TMPD + cytochrome C Oxidase -----\> TMPD is OXidized ( loss of electron)
161
TMPD turns what color when it is oxidized and loses an electron?
Blue
162
three possible outcomes for Oxidase test?
Oxidase Negative
Oxidase positive
False positive
163
what do the test results mean
Oxidase Negative
TmPD on Window
No cytochrome C Oxidase
No Oxidation of TMPD
No color change
164
what do the test results mean?
Oxidase Positive
TMPD on Window
Cytochrome C Oxidase
Oxidation of TMPD
Blue
165
what do the test results mean?
False Positive
TMPD is unstable
and can lose its electron
with or without Cytochrome C
Oxidase
to prevent test must be read in 20-30 secounds
166
what does the nitrate reduction test tell us?
determins is the organisum can use nitrate for denitrification or ammonification.
167
Aerobic Respiration
aerobic and facultative aerobic organisms
O2 is the final electron acceptor
168
Anaerobic Respiration
Nonoxygen molecule is final electron acceptor.
169
If ______ is used as the final electron acceptor then the bacteria can perform either.
NO3
170
\_\_\_\_\_\_ is a multiestep process performed by a group of enzymes that leads to the production of N2 (Nitrogen Gas)
Denitrification
171
\_\_\_\_\_\_\_ is a multistep process performed by a group of enzymes that lead to the production of NH4(ammonia)
Ammonification
172
what is the pricipal of the Nitrate reduction test?
1st step in either denitrification or ammonification is the reduction of nitrate (No2) to Nitrite ( No2) by the enzyme Nitrate Reductase.
173
Nitrate Broth contains
Potassium Nitrate ( KNO3) - source of Nitrate
Duraham Tube - to monitor gas production
there are no color indicators in this test
174
three possible outcomes for the Nitrate Reduuction Test are?
first you need to know the results of the OF test or Phenol Red Broth test is the organisum a Fermenter or Non fermenter.
1. organisum is a Non fermenter
2. organisum is a fermenter.
3. organisum is a fermenter
175
Nitrate reduction test results
Organisum is a Non Fermenter
Gas-Yes
Nitrate reductase present
Denitrification- source of gas
Nitrate Reductase Positive
176
Nitrate reduction test results
Organisum is a Fermenter
Gas-yes-----------\> type of gas is unknown and test must proceed
Addition of Reagent A
Addition of Reagent B
Red color
Nitrate Reductase Positive ---------\> Nitrate ( NO3) converted into nitrate ( NO2) which reacted with water in broth to form (HNO2) nitrous acid which Reagent A & B reacted with.
177
Nitrate reduction test results
Organisum is a Fermenter
Gas--yes Or No
Addition of Reagent A
Addition of reagent B
No Red Color-----\> possibility #1 orgnisum does not have Nitrate Reductase
----------\> possiblity # 2 .. organisum reduced all nitrate to nitrite and all the nitrite was converted to other nitrogen compounds.
After adding Zinc powder
red color----\>nitrate still present---\>Nitrate Reductase Negative
No Red Color---\>no Nitrate present--\> Nitrate Reductase Positive
178
what does the Decarboxylation Test tell us?
Determins the presence or absence of a specific Decarboxylase in a bacterium.
179
Is an enzyme that catalyzes the removel of an amino acids carboxyl group.
Decarboxylase
it acts on specific ameno acids
End products of decarboxilation are Polyamines ( compunds with 2 or more amine groups.)
A co enzyme is required called Pyridoxyl Phosphate
180
are important in maintaining the correct internal PH of a bacterial cell.\_\_\_\_\_\_\_\_ are alkaline compounds that raise \_\_\_\_\_
POLYAMINS
Polyamines
PH
181
other importances of Polyamines
Regulation of gene transcription and translation
role in biofilm formation
182
Principal of the Decarboxylation Test.
Mollers Decarboxylase Base medium contains :
protiens ( beef extract and peptones
Carbohydrates( glucose)
Pyridoxal Phosphate (co enzyme)
Ph indicator ( bromocresol Purple)
183
Principal of the Decarboxylation Test.
----Also specific amino acids are added to test for corresponding decarboxylasae.
HCL is added so that the initial PH is 5.2-6.2 since decarboxylases are induced /made under acidic conditions.
After media is inoculated a layer of oil is added to make an anaerobic enviroment since it stimulates fermentation( and acide production which futher lowers PH)
184
three possible outcomes for Decarboxylation Test.
1. decarboxylase Negative- no fermentation
2. Decarboxylase Negative- fermentation
3. Decarboxylase Positive.- fermentation
185
Results for Decarboxylation Test
Decarboxylase Negative
no fermentation
No acid produced = PH
No decarboxylation induced
no alkaline products = PH
No color change ( copper)
186
Decarboxylase Negative
fermentation
Acid produced ( lower PH) yellow
No decarboxylation induced.
No alkaline products = PH
Tube remains Yellow
187
Decarboxylase Positive
Fermentation
Acid produced induced
decarboxylation induced
alkaline products produced ( raises PH)
tube turns purple
188