Practical Sessions Flashcards
Susceptibility of a microrganism to anitbiotics cannot…
Be predicted sometimes. This is can be due to the resistance that they form towards antibiotic
Why do microrganisms form resistance towards antibiotics?
a) Exposure of the microbe to many agents especially micro-flora that reside in the body
Drugs selcted to treat patients must be….
a) must be effective in treating
b) have less impact of the patient
c) should not disruot the microflora of the body
A laboratory test done to determine the effectiveness of the antibiotic administeration against specific pathogens
Antibiotic Sensitivity Test
Why is antibiotic sensitivity testing done?
a) Allow control of antibiotic use in clinics
b) Helps with the knowledge of which drug to prescribe
c) Allows one to view changes in antibiotic therapy of certain microbes (resistance)
In which cases should antibiotic senstivity test be performed?
a) When the patient is immunodeficient therefore can contract to many pathogens
b) When the condition being treated can cause death
c) When the pattern of antimicrobial sensitivity is unknown
d) To check resistance patterns of microbes to specific drugs
In which cases is antibiotic sensitivity test not perfomed?
a) When the pathogen has a known sensitivity pattern
b) When there are mixed cultures therefore contamination is present
What is a disadvantage of the E-test?
Is that individual test strip are needed for each antibiotic thus can be very costly
What is the deal with the Broth Dilution Method?
It is beneficial in the determinning of the minimum inhibitory concentration of the antibiotic
It’s disadvantages is that it will be time consuming in the sense that one has to expose the microbe to varing amounts of antibiotic one at a time
What is the limit of the Disk Diffusion?
It cannot be used to test slow growing bacteria
Why is there a limitation in the Antibiotic Sensitivity Test?
It is because it doesn’t reflect the same sensitivity because the patient’s exposure to the pathogen maybe extensive in terms of development of new ways in which the pathogen evades the immune system
In the identification of bacteria what does one start with?
Gram staining and then observing the shape of the bacteria either cooci or rod
pH indicators serve what purpose?
They are used in fermentation reaction where there is change of colour depending on the fermentation - acidic or alkaline
Glucose, lactose, …. and …. are used in carbohydrate fermentation
Mannitol and maltose
Which agar utilize carbohydrate fermentation?
- MacConkey Agar
- Mannitol Salt Agar
- Triple Sugar Iron Agar
- Mueller Hinton Agar
Why is the Mannitol Salt Agar a selective and differential test?
Because it consists of NaCl which is used to only allow for the growth of Staphylococcus spp
It is differential because it consists of mannitol which is a sugar that Staphylococcus aureus can ferment. So it allows for differentiation of S. aureus and other S spp
The MacConkey Agar consists of which carbohydrate?
Lactose
How can one differentiate lactose fermentators to non lactose fermenters?
Lactose fermenters : They produce lactic acid therefore causing the decrease in pH and cause a change in colour: They form Red colonies
Non lactose fermenters: They do not produce lactose therefore no change in colour : they from cream colonies
What is the function of the catalase enzyme?
It catalyzes the breakdown of hydrogen peroxide to water and O2.
For the catalase test how would one know that the test is + or - ?
+ Catalase = formation of bubbles
- Catalase = no bubbles are formed
In the catalase test what can produce a false positive?
Enterococci, consists of peroxidase enzyme which can catalyse H2O2 to H2O and Oz but at a much slower rate
What is the purpose of the coagulase test?
It is used to differentiate Staphylcoccus aureus from other Staphylococci spp
S aureus produces two forms of coagulase, what are they?
Bound and Free
What is bound coagulase?
An enzyme located on the cell wall of the S aureus and it reacts with fibrinogen directly. After reaction is causes the fibrinogen to attach itself onto the wall of the S aurerus thus causing it to clump when mixed with plasma
What is free coagulase?
It is an outside of the cell protein that causes clumping when S aureus is placed with plasma
What coagulase enzyme causes activation of plasma coagulasee reacting factor (CRF), which is then altered to form coagulase CRF complex?
Free coagulase
How would one know the coagulase test is +?
There will be clumping in less than 10 seconds
When the coagulase test comes back -, how would one confirm 100% that it is -?
Use the tube test.
Explain the concept of the tube test?
It is used to for confirming if the coagulase test is really negative.
If the test is negative after 4 hrs then place it again for another 24 hrs
if the - then no clots form
Describe the mechanism of the DNAase test?
It is used to differentiate S aureus from other S species
It works in such a way that pathogen consists of enzymes known as DNAses that break down DNA in the presence of HCl in order to use the DNA for energy and growth
If the DNAse test is + then….
There are clear zones formed
Functions to
a) Differentiate S. agalacataie from other Streptococci species
b) Used to identify L. monocytogenes
The CAMP test
Wht does CAMP factor do?
It is used to enlarge the area of hemolysis formed
Novobiocin Sensitivity Test
It is used to differentiate coagulase - Staphylococcus
It is used to identify S. epidermis from S. saprophyticus
Has a clear zone more than 12 mm is…
Novobiocin sensitive
Staphylococcus epidermis