Kirby-Bauer test and
Endospore Staining
Kirby-Bauer or Disk Diffusion Test
• Designed to test the effectiveness of antibiotic
agents and chemicals at inhibiting bacterial
growth
• Uses Mueller-Hinton agar, allows agent to
diffuse freely from disk
• Primary test preformed in clinical laboratories
with strict quality control
Materials
•
•
•
•
•
Petri plates with isolated colonies
Petri plate with Mueller-Hinton agar
Sterile cotton swab
Sterile DI water
Dispenser, antimicrobial disks and/or disks
impregnated with other agents
• Forceps
• Alcohol
• Ruler
Method
• Soak tip of sterile cotton swap in sterile water
• Pick up single colony from designated plate
• Swab entire target plate with cotton swab, make
sure to crisscross the plate multiple times from
multiple directions to ensure full coverage
• Allow plate to sit for 5 minutes
• Using sterile forceps (via microincinerator) Place
impregnated disks evenly spaced around petri
plate
• Gently tap the disk to ensure full contact with
agar
• Write a code for each of the disks on the bottom
of the petri plate to reference results and records
• Invert and incubate
Recording
• Use the table below as an example to record
results
Bacterium 1
Agent
Code
Zone
Size
Results
Bacterium 2
Zone
Size
Results
1.
2.
3.
4.
5.
6.
7.
*S – susceptible; I – Intermediate; R – Resistant
Bacterium 3
Zone
Size
Results
Interpretation of Zones
Report Format
•
Title: Top of 1st page
•
Authors: list authors in alphabetical order
•
Abstract: a brief summary of the experiment and results
•
Material and Methods: Describe the actions taken in each
step, formulas used both math and media.
•
–
Break down by steps i.e. media prep, inoculation, cell counts
etc…
–
Include tables or figures as needed to summarize data sets like
nutrient targets and masses used.
Cross-Reference: When including tables and figures make
sure to cross reference them in the document where they
are mentioned i.e. “the target concentrations of the
media can be found in Table 1”, or nutrient mass were
weight out (Table 1) on a MT SI-114 balance”
Report Format
•
•
•
Results: Keep same formatting as material and
methods, but describe the results from each step.
–
Do not make conclusions or explain why the result was
observed
–
Include cross referenced figures/tables at end of section
Conclusions: Explain why your results came out as they
did. Make observation referencing your results, and
hypothesize what could be done to change your results
–
If odd things need to be explained at each step then break
down by sections
–
If everything went according to materials and methods then
a paragraph is fine
References: If any references are used as a descriptor
for a technique used or to compare results include
them here.
Schaeffer-Fulton Endospore Staining
• Spores are resistant due to the protein keratin
that makes up the outerlayer
• Keratin is also resistant to stains
• Uses Malachite green to stain the spore
• Malachite green is water soluble and has low
affinity for cellular material i.e. washes out of
bacterial cell walls but not spore cell walls
Method
• Prepare a smear of
Bacillus and allow to air
dry
• Heat fix the smear by
holding back side of slide
against microincinerator
mouth
• Flood the heat smear
with malachite green and
allow to stand for 5
minutes then apply heat
for 10 second
• Wash smear with gentle
stream of water
• Counterstain with
safranin for 45 seconds
Purchase answer to see full
attachment
Why Choose Us
- 100% non-plagiarized Papers
- 24/7 /365 Service Available
- Affordable Prices
- Any Paper, Urgency, and Subject
- Will complete your papers in 6 hours
- On-time Delivery
- Money-back and Privacy guarantees
- Unlimited Amendments upon request
- Satisfaction guarantee
How it Works
- Click on the “Place Order” tab at the top menu or “Order Now” icon at the bottom and a new page will appear with an order form to be filled.
- Fill in your paper’s requirements in the "PAPER DETAILS" section.
- Fill in your paper’s academic level, deadline, and the required number of pages from the drop-down menus.
- Click “CREATE ACCOUNT & SIGN IN” to enter your registration details and get an account with us for record-keeping and then, click on “PROCEED TO CHECKOUT” at the bottom of the page.
- From there, the payment sections will show, follow the guided payment process and your order will be available for our writing team to work on it.