Lab Pract. 2 Flashcards
Explain the function of CARBOHYDRATES in broth media.
1.) They are substrates for fermentation (yellow) ( A for acid).
Explain the function of PEPTONES in broth media.
- ) Nutrient source
- ) When peptides break down, ammonia is released and will change color of the broth
- ) Alternative substrate for non-fermenters (hot pink) ( K for alkaline)
Explain the function of PHENOL RED in broth media. What is it made of?
A pH indicator:
- ) Orange/pink at neutral pH 7
- ) Hot pink above 7.4
- ) Yellow bellow pH 6.8
- ) Allows us to determine whether or not an acid has been formed as an end product of fermentation.
5.) Made of peptone (hydrolyzed proteins), carbohydrate, water, phenol red
Explain the function of DURHAM TUBE in broth media.
- ) Turned upside down in broth
2. ) Used to collect gas that is being released by the organism during fermentation
What are the end products of aerobic respiration? Are any of these products acidic? How do they know if the cultures are using aerobic respiration or fermentation for their energy production?
End products:
- ) Water
- ) ATP
- ) Carbon Dioxide
1.) Fermentation pH will drop below 6.8 and turn broth yellow , Aerobic respiration is orange
Do all the organisms use pyruvic acid in the same way for fermentation?
1.) No, metabolism of pyruvate is not the same for all microorganisms. An assortment of end products define their fermentative capabilities
How would you interpret your results if you inoculated a carbohydrate broth tube and after incubating the tube for several days, the broth had turned hot pink in color?
1.) Broth is negative for fermentation and are “K” (alkaline).
How do pH indicators work?
- ) Weak acids that reflect a particular wavelength of light
- ) Placed in solutions of different pH’s, the loss or gain of electrons causes them to absorb a different wavelength of light
- ) Slight chemical change causes them to reflect different wavelength (color) of light then previous
If a particular bacterium is forming acids from a specific carbohydrate during fermentation, the pH will?
- ) Drop below 6.8 and turn broth yellow
2. ) Report this as A (acid)
Some bacteria genera do ferment carbohydrates to form acids, but can utilize the peptone present in the broth as a nutrient source. When these peptides are broken down?
1.) Ammonia is released
Ammonia, which has a pH of 10 will turn a broth what color?
1.) Hot pink (negative for fermentation/reported as K for alkaline)
Broths that remain orange/pink are considered?
1.) Negative for fermentation
Many phenol red broths will have a small tube turned upside down in the broth. What is this called and what does it do?
- ) Durham tube
2. ) Used to collect gas that’s being released from organism during fermentation
When inoculating broth that contain Durham tubes, you should not swish your loops violently b/c this introduces air bubbles into the tube. If your’e careful when inoculating your broth any glass that accumulates in the tube will be interpreted as?
- ) Positive result
2. ) Reported as (G for gas)
An isolate that is turning phenol red broth yellow and releasing gas, which is collecting gas in the Durham tube will be reported as?
1.) AG
(Fermentation pathways)
Escherichia/Shigella begins with and ends with what products?
- ) Begins w/ pyruvate as final electron acceptor
2. ) Ends w/ mixed acids
(Fermentation pathways)
Proteus begins w/ and ends w/ what products?
- ) Begins w/ pyruvate as final electron acceptor
2. ) Ends w/ CO2 and H2
(Fermentation pathways)
Acetobacter begins w/ and ends w/ what products?
- ) Begins w/ pyruvate as final electron acceptor
2. ) Ends w/ Acetic acid
(Fermentation pathways)
Propionibacterium begins w/ and ends w/ what products?
- ) Begins w/ pyruvate as final electron acceptor
2. ) Ends w/ Proprionic acid
(Fermentation pathways)
Enterobacter/Klebsiella/Serratia begins w/ and ends w/ what products?
- ) Begins w/ pyruvate as final electron acceptor
2. ) Ends w/ Butanediol (an alcohol)
Describe reversion.
- ) Due to incubation time, once a bacteria has exhausted the supply of available carbohydrate, they will sometimes start digesting peptone
- ) The medium will reflect an increase in pH w/ a color change from yellow back through orange to hot pink
What would happen if powdered zinc was added to an uninoculated nitrate broth tube?
- ) Catalyze any remaining NO3 (nitrate) to NO2 (nitrite)
2. ) NO2 (nitrite) will combine w/ sulfanilic acid and will form red precipitate
How does nitrogen reducing soil bacteria participate in the nitrogen biogeochemical cycle?
- ) “Fix” nitrogen and convert nitrogen to ammonia NH3
- ) Helps plants get nutrients from air and soil
3) Plants don’t have enzymes to convert Nitrogen into a biologically useful form
In step 2, your P. aeruginosa tube remained clear, indicating that neither NO3- nor NO2- were present in the tube. What happened to the NO3- that was present in the tube before you inoculated your tube?
1.) It gassed out of the tube in the form of NH4+ or N2
Why is it advantageous for enteric bacteria to perform anaerobic respiration?
1.) Allows them to live in environments where the oxygen is low to nonexistent
(Motility of bacteria)
S. typhimurium
Motile (+)or(++) or Non-motile (-):
1.)
Using an ETC? (+) (-):
1.)
(Motility of bacteria)
P. Vulgaris
Motile (+)or(++) or Non-motile (-):
1.)
Using an ETC? (+) (-):
1.)
(Motility of bacteria)
S. aureus
Motile (+)or(++) or Non-motile (-):
1.)
Using an ETC? (+) (-):
1.)
(Motility of bacteria)
M. smegmatis
Motile (+)or(++) or Non-motile (-):
1.)
Using an ETC? (+) (-):
1.)
When observing the results of the motility test, why do aerobes sometimes show inconclusive results?
- ) They are obligate aerobes
- ) Can only survive near the surface of the tube
- ) Inconclusive for motility, positive for aerobic respiration
What is indicated when the bacteria growing along the stab do not turn red?
1.) Bacteria are producing ATP via fermentation
List two findings that can be determined about your unknown from the results of this medium.
- ) Ability to move through watery medium
2. ) Respiratory status
During aerobic respiration________serves as the final electron acceptor? In this catabolic pathway, oxygen bonds w/ two hydrogen ions to form water, an event which is catalyzed by?
- ) Oxygen (catabolic)
2. ) Cytochrome oxidase
Describe cytochrome oxidase.
- ) An enzyme also known as Complex IV
- ) Large transmembrane protein complex found in bacteria and the mitochondrion of eukaryotes.
- ) Last enzyme in the respiratory electron transport chain of mitochondria or bacteria located in the mitochondrial or bacterial membrane.
What is the key difference between aerobic and anaerobic respiration during anaerobic respiration?
1.) An inorganic molecule such as nitrate, nitrite or sulfate serves as an alternate final electron acceptor, rather than oxygen (which is also inorganic)
What is the benefit of using anaerobic respiration as an alternate final electron acceptor even though it generates less ATP for microorganisms?
- ) Allows them to survive in a greater diversity of environments (dry and waterlogged soils)
- ) Environments w/ O2 diffusion may be slow or nil
What’s the benefits of using the motility test?
- ) Allows us to differentiate between species
- ) Its ability to move through a watery medium
- ) Respiratory status of the isolate (is the bacteria using a respiratory chain when producing ATP
As the nutrient level drops,__________bacteria will remain in place, but the___________bacteria will remove away from the stab line in search of nutrients.
- ) Non-motile bacteria
2. ) Motile bacteria
In many motility media, an indicator dye-Triphenyltetrazolium trichloride (TTC) is incorporated into the medium to?
- ) Allow for easier visibility of motile bacteria
2. ) Gives info regarding respirator status of organism
TTC (Triphenyltetrazolium trichloride) is a colorless molecule that is a?
1.) Competitive inhibitor of Complex 1 of the ETC (also called NADH dehydrogenase).
During respiration in the bacterial cell, NADH donates_______to Complex I?
1) Electrons
If TTC is present in the bacterial cell, NADH can donate an electron pair to TTC instead of donating them to?
1.) Complex I
What happens when NADH donates an electron pair to TTC? What happens to the color of the bacteria using this electron transport chain?
- ) TTC is reduced to formazan, a red colored molecule.
2. ) Bacteria turn red
Bacteria that are producing ATP via fermentation will remain?
1.) Colorless