2. With reference to the table below, determine the antibiotic susceptibilities of different antibiotics against E.coli and S.aureus. E.coli Antimicrobial agent Ampicillin Tetracycline S.aureus Antimicrobial agent Tetracycline Interpretive categories and zone diameter breakpoints, nearest whole mm S ≥17 ≥15 I 14-16 12-14 S ≥19 R ≤13 ≤11 (Reference: CLSI 2018) Interpretive categories and zone diameter breakpoints, nearest whole mm 15-18 R <14

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For a laboratory exercise of antibiotic susceptibility test in a disk-diffusion method. With reference to the table below, determine the antibiotic susceptibilities of different antibiotics against E.coli and S.aureus.
2. With reference to the table below, determine the
antibiotic susceptibilities of different antibiotics
against E.coli and S.aureus.
E.coli
Antimicrobial
agent
Ampicillin
Tetracycline
S.aureus
Antimicrobial
agent
Tetracycline
Interpretive categories and zone diameter breakpoints, nearest
whole mm
S
≥17
≥15
I
14-16
12-14
S
≥19
R
<13
≤11
(Reference: CLSI 2018)
Interpretive categories and zone diameter breakpoints, nearest
whole mm
I
15-18
R
<14
(Reference: CLSI 2018)
Transcribed Image Text:2. With reference to the table below, determine the antibiotic susceptibilities of different antibiotics against E.coli and S.aureus. E.coli Antimicrobial agent Ampicillin Tetracycline S.aureus Antimicrobial agent Tetracycline Interpretive categories and zone diameter breakpoints, nearest whole mm S ≥17 ≥15 I 14-16 12-14 S ≥19 R <13 ≤11 (Reference: CLSI 2018) Interpretive categories and zone diameter breakpoints, nearest whole mm I 15-18 R <14 (Reference: CLSI 2018)
Materials:
Broth cultures of Staphylococcus aureus and
Escherichia coli
• Mueller-Hinton agar plates
. Antibiotics (Ampicillin & Tetracycline)
• Sterile Filter Paper disks
• Sterile Cotton Swabs
• Forceps
Procedure:
1. Label two agar plates with group number, date, name of
the organism used along the edge of the plate. Divide
the bottom of the plates into two sections and label
each section with the name of antibiotic used (AM-
Ampicillin; Te Tetracycline). Duplicates for each
type of bacteria.
2. Dip a sterile cotton swab in the bacterial suspension
being tested. Squeeze the swab against the inner wall
of the tube to remove excess liquid.
3. Swab the entire plate from top to bottom, edge-to-edge
leaving no gaps. Rotate the plate and using the same
swab, again swab the entire plate from top to bottom.
Allow the plates to dry for five minutes.
4. Sterilize the forceps by placing in alcohol. Flame the
forceps to remove the alcohol. (Once they have been
sterilized, do not touch the forceps' tips to anything.
If the tips touch anything, re-sterilize them.)
5. Use the sterile forceps, pick up a filter paper disk and
place on the empty petri dish.
6. Add 20 µL of antibiotic solution being tested on the
center of the paper disk.
7. Using the sterile forceps, carefully place the moist disk
in the center of the appropriate section of the plate
inoculated with S. aureus. Gently press the paper disk
onto the surface of agar.
8. Repeat steps 4-6 with other antibiotics.
9. When you have placed antibiotic paper disks on all
sections of the plates, invert them and incubate the
plates at 37°C for 24
10. Repeat steps 2-8 with E. coli.
11. After incubation, measure the diameter of the zone of
inhibition (mm). Examine the plates carefully for
well-developed colonies within the zone of inhibition.
12. Compare the susceptibility of bacterial cultures to
antibiotics tested.
apap
EDED
Group Ane Date
Figure 5.1 Antibiotic susceptibility test.
Transcribed Image Text:Materials: Broth cultures of Staphylococcus aureus and Escherichia coli • Mueller-Hinton agar plates . Antibiotics (Ampicillin & Tetracycline) • Sterile Filter Paper disks • Sterile Cotton Swabs • Forceps Procedure: 1. Label two agar plates with group number, date, name of the organism used along the edge of the plate. Divide the bottom of the plates into two sections and label each section with the name of antibiotic used (AM- Ampicillin; Te Tetracycline). Duplicates for each type of bacteria. 2. Dip a sterile cotton swab in the bacterial suspension being tested. Squeeze the swab against the inner wall of the tube to remove excess liquid. 3. Swab the entire plate from top to bottom, edge-to-edge leaving no gaps. Rotate the plate and using the same swab, again swab the entire plate from top to bottom. Allow the plates to dry for five minutes. 4. Sterilize the forceps by placing in alcohol. Flame the forceps to remove the alcohol. (Once they have been sterilized, do not touch the forceps' tips to anything. If the tips touch anything, re-sterilize them.) 5. Use the sterile forceps, pick up a filter paper disk and place on the empty petri dish. 6. Add 20 µL of antibiotic solution being tested on the center of the paper disk. 7. Using the sterile forceps, carefully place the moist disk in the center of the appropriate section of the plate inoculated with S. aureus. Gently press the paper disk onto the surface of agar. 8. Repeat steps 4-6 with other antibiotics. 9. When you have placed antibiotic paper disks on all sections of the plates, invert them and incubate the plates at 37°C for 24 10. Repeat steps 2-8 with E. coli. 11. After incubation, measure the diameter of the zone of inhibition (mm). Examine the plates carefully for well-developed colonies within the zone of inhibition. 12. Compare the susceptibility of bacterial cultures to antibiotics tested. apap EDED Group Ane Date Figure 5.1 Antibiotic susceptibility test.
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