antimicrobial technique
TRANSCRIPT
![Page 1: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/1.jpg)
MICROBIOLOGY
ANTIMICROBIAL TECHNIQUE
MEMBERS :IMRAN KHAN SULAIMAN
CHE ABDUL HALIMMUHD HUZAIFAH
NURUL ATIQA IBRAHIMNADHIRAHTUL HUSNA
NOR ZAFIFAHROSE AMALINA
![Page 2: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/2.jpg)
• kills or inhibits the growth
of microorganism such
as bacteria , fungi, or protozoa
What is antimicrobial ?
![Page 3: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/3.jpg)
kill microbes
(microbiocidal)
prevent growth of microbes
(microbiostatic).
![Page 4: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/4.jpg)
antimicrobial substances used on non-living objects or
outside the body.
![Page 5: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/5.jpg)
Why antimicrobial technique is needed ?
• Bacteria have the ability to develop resistance following repeated or subclinical (insufficient) doses
• so more advanced antibiotics and synthetic antimicrobials are continually required to overcome them.
![Page 6: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/6.jpg)
![Page 7: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/7.jpg)
Technique in testing for Antibiotic sensitivity
• The method includes several steps including :• obtaining a bacterial sample• identifying the type of bacteria in the bacterial
sample• selecting a set of antibiotics based on the
identity of the bacteria in the bacterial sample• obtaining a control sample from the bacterial
sample
![Page 8: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/8.jpg)
Antimicrobial technique . . .
![Page 9: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/9.jpg)
Antimicrobial agent placed on the surface of an
agar plate
medium that has been inoculated with the disease
agent being tested, which will grow and
fill the disk.
killing some of the disease agent around where the
anitmicrobial agent was
innoculated
disease agent shows how
effective the antimicrobial
agent is.
It containing
Antimicrobial agent diffuse into medium
size of the area cleared
Disk diffusion
![Page 10: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/10.jpg)
For example . . .
• Small wafers containing antibiotics are placed onto a plate which bacteria are growing.
• If the bacteria are sensitive to the antibiotic, a clear ring, or zone of inhibition, is seen around the wafer indicating poor growth
Antibiotic sensitivity testing
![Page 11: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/11.jpg)
Zone sizes differ on sensitivity pattern
• It has been determined that zones of inhibition of a certain diameter (varies for antibiotic and to a lesser extent, bacterial species) correlate with sensitivity or resistance to the antibiotic tested
![Page 12: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/12.jpg)
Steps for disk difussion method
![Page 13: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/13.jpg)
• to determine whether or not a specific strain of bacterium or fungus is susceptible to the action of a specific antibiotic.
• commonly used in the setting of medicine, where a particular organism has been found to infect a patient, and the doctor treating the patient is seeking guidance on what concentration of antibiotic is suitable.
Epsilometer test
![Page 14: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/14.jpg)
rectangular strip that has been
impregnated with the drug
A lawn of bacteria is spread and
grown on an agar plate
Etest strip is laid on topinto the agar
an exponential gradient of the drug
to be tested
Etest utilise by then
Drug diffuse out
andproducing
Epsilometer test
![Page 15: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/15.jpg)
As a result . . .
• an exponential scale printed on the strip .
• After 24 hours of incubation, an elliptical zone of inhibition is produced
• the point at which the ellipse meets the strip gives a reading for the minimun inhibitory concentration (MIC) of the drug.
![Page 16: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/16.jpg)
Dilution plating technique
• to reduce the concentration of microscopic organisms or cells in a sample
• A series of cultures is tested with various concentration pf an antibiotic to determine minimum inhibiting concentration of antibiotic
![Page 17: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/17.jpg)
spread bacteria over a wide area
count the number of colonies that grow
bacterial cell in the original sample should produce a single colony
The numbers of bacteria colonies that grow on
each plate are counted
![Page 18: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/18.jpg)
Stokes’ Method• In original Stokes’ method
the inoculum of the control strain is evenly spread over the upper and lower thirds of a plate
• the test strain over the central third,uninoculated (gaps 2 – 3 mm wide are left to test from the control areas.
![Page 19: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/19.jpg)
For example . .
• In the Stokes controlled sensitivity test, a control organism is inoculated on part of a plate
• the test organism is plated on the remainder.
• Disks are placed at the interface and the zones of inhibition are compared.
• The use of a sensitive control shows that the antibiotic is active
• if the test organism grows up to the disk it may safely be assumed that the test organism is resistant to that drug.
![Page 20: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/20.jpg)
Testing Minimum Inhibitory Concentration
• In alternative measure of susceptibility is to determine the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) of a drug.
• Both are mixed with serially diluted antibiotic solutions and a standard inoculum is applied.
• After incubation, the MIC is the first broth in which growth of the organism has been
![Page 21: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/21.jpg)
What is Minimum Inhibitory
concentration• Minimum inhibitory concentration (MIC), in is the lowest concentration of an antimicrobial that will inhibit the visible growth of a micro organism after overnight incubation.
• Minimum inhibitory concentrations are important in diagnostic laboratories to confirm resistance of micro organisms to an antimicrobial agent and also to monitor the activity of new antimicrobial agents.
![Page 22: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/22.jpg)
The Antibiotics are diluted to various dilution to test the minimum inhibitory concentration
![Page 23: antimicrobial technique](https://reader035.vdocuments.us/reader035/viewer/2022062312/5570188ed8b42ac0178b538d/html5/thumbnails/23.jpg)