Microbiology Exercise 14
CHAPTER 14
To make sure bacteria culture is present
Why is it important to record the presence of growth?
allows time for fermentation
incubate the tubes at 35 C for 48 hrs. or longer. Why is the time of incubation important.
It produces ammonia which raises the PH and turns the agar red. This is done aerobically so only the top portion will be red.
What happens when the organism uses peptones in the agar?
The organism needs time to ferment the glucose otherwise you could get a false postive and the tube would be all yellow.
Why would you not analyze results too early?
If all the sugar get use up when all the peptones are used up it would turn back to red giving you a false negative.
Why not analyze the results too late?
agree, if a bacterium cannot ferment glucose which is a simple sugar, it cannot ferment lactose either because lactose is a complex
a bacterium that doesnt ferment glucose will not ferment lactose.
red (neutral)
color of uninoculated medium
E.coli produces large amounts of acid from pyruvic acid (pH < 5.5)
En. aerogenes makes acetoin (pH > 6.0 neutral)
E.coli and Enterbacter aerogenes are small gram-negative rods. How can you differentiate them?
fermentation occurs 1-24 hrs
fermentation that results in acid production will turn the indicator yellow (pH of 6.8 or below)
prolong incubation periods greater then 24 hrs, bacteria will begin growing oxidatively on the peptone after using up the carbohydrate supplied
causing neutralization of the indicator and turning it red because of ammonia production.
Why are fermentation tubes evaluated at 24 and 48 hrs?
the organism would oxidase peptone and produce ammonia resulting in turning the indicator from yellow(production of acid) to red (neutral)
What would happen if an organism used up all the carbohydrates in a fermentation tube.
gas will be trapped in the inverted or Durham tube. Bubbles will appear
if an organism metabolizes glucose aerobically, what result will occur in the fermentation tubes?
is a differential media, used to distinquish organisms that produce large amounts of acid from glucose and organisms that produce the neutral product acetoin
large amount of acid from glucose + methyl red = red (MR positive; pH is below 4.4)
if neutral products are produced + methyl red = yellow (pH above 6.0)
production of acetoin is detected by the addition of potassium hydroxide and a-naphthol = upper part of the medium will turn red
negative V-P test = light brown
MRVP test
is a selective media, allows the growth of certain type of organisms, while inhibiting the growth of other organisms.
For example, organisms that have the ability to utilize a given sugar are screened easily by making that particular sugar the only carbon source in the medium for the growth of the microorganism
Fermentation Tubes
Simmon's Citrate Agar Slant (SC)
CHAPTER 25
Bacteriostatic, until we can perform a subculture to determine if it is bacteriocidal.
is the disk-diffusion technique measuring bacteriostatic or bacterialcidal activity? Explain
During the log phase when the cells are dividing exponentially
In which growth phase is an organism most sensitive to an antibiotic?
Other factors contribute to performance in vivo. These include, variations in PH, patient nutrition, side effect, other medical conditions, additional drug interactions.
Why is the disk-diffusion technique not a perfect indicationof how the drug will perform in vivo? What other factors are consdered before using the antimicrobial agent in vivo?
tetracycline would inhibit effectiveness of PCN (no reproduction) PCN needs bacteria to grow to inhibit cell wall synthesis, but tetracycline is a bacteriostatic
what effect woiuld presence of tetracycline in the body have on penicillin therapy?
Each antimicrobial agent inhibits growth differently by:
Cell wall structure
Permeability
Species
Why isn't one antimicrobial agent equally effective against all three bacteria?
A=1:100
B=1:160
Broth dilution: What is the minimum bactericidal concentration of each antibiotic?
A=1:80
B=1:150
Broth dilution: What is the minimum bactericidal concentration of each antibiotic?
B
Which antibiotic is more effective against Staphylococcus aureus?
N/A
Which antibiotic is more effective against Salmonella enterica?
if its a salt tolerant organism or mannitol-fermenting organism
After you have observed a gram-pos coccus, what is the additional info you need before perfoming a coagulase test?
It is both a selective and differential medium. The salt concentration doesn't let some bacteria grow. The mannitol component is used for differential analysis for microorganisms that ferment mannitol
why is mannitol salt agar used as a selective medium for normal skin microbiota?
gram-positive
salt tolerant
produce coagulase
list three characteristics of Staphylococcus aureus
dry layer of epidermis
sebum
salt;hypertonic environment
list the three factors that protects the skin from infection
coaguluase is an enzyme that coagulates (clots) the fibrin in blood
cell bound coagulase could provide an antigenic disguise if it clotted fibrin on the cell surface
could make the bacterial cells resistant to phagocytes or tissue bactericides or even drugs which might be unable to diffuse to their bacterial target
what is coagulase? how does it relate to pathogenicity?
disk-diffusion method
assume that you isolated S. aureus from your skin. How would you determine whether it is penicillan-resistant?
CHAPTER 46
S. pneumoniae cannot be differentiated from other alpha-hemolytic streptococci on blood agar.
optochin inhibition is used to identify this pathogen; inhibition zone greater then 15mm or larger indicates optochin sensitivity
subculturing to a blood agar plate and placing bacitracin and optochin disks on the area subcultured will help determine whether pathogenic streptococci is present. Explain