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Test tube Nutrient broth Culture of micro- Antibiotic: ciprofloxacin. (In Active salt (final concentration Antibiotic: ciprofloxacin (Final concentration in Turbidity observed
no. media (ml) organism from teeth microgram / ml, Final in % w/v) microgram / ml) with final concentration of 0.05%
sample (in micro liter) concentration) w/v Active salt
Ciprofloxacin Cipro+NaCl
TURBIDITY
MIC[Cipro+NaCl]
MIC[Cipro]
0
0 2.5 5 20 40 60
CIPROFLOXACIN CONC. ( G/ML)
DISCUSSION AND CONCLSION OF MIC
Since it was investigated and reported that any kind of biofilm made by the
microorganisms exhibit a marked decrease in susceptibility to antimicrobial agents and
host defence systems compared to their planktonic counterparts, our main investigation
was to study on measure of degree of decreased susceptibility to certain antibiotics by
the biofilm producing microorganisms. The minimum inhibitory concentration and the
zone of inhibition study were performed using different parameter to establish that.
the MIC for only Ciprofloxacin is 40 micrograms/ml and for the combination with Sodium
Chloride, which acts as an active salt, is 5 micrograms/ml. i.e., susceptibility to combined
treatment is 8 times more potent than that of only Ciprofloxacin for the biofilm making
microorganisms.
only salt is not sufficient to reduce the growth of the microorganism. Which clearly
indicates that when bactericides are combined to a salt solution, they are capable of
killing microorganisms which are retrieved as planktonic organisms and no longer
organized as a biofilm due to the salt effect.
ZONE OF INHIBITION
If an antibiotic stops the bacteria from growing or kills the
bacteria, there will be an area around the drug or antbiotic
where the bacteria have not grown enough to be visible.
This is called a zone of inhibition.
The size of this zone depends on how effective the antibiotic is at
stopping the growth of the bacterium.
PRACTICAL WORK
Materials required:
1. Culture of microorganism from teeth sample.
2. Culture of Pseudomonas aeruginosa.
3. Nutrient broth.
4. Antibiotic: ciprofloxacin.
Preparation of nutrient agar media
Preparing agar plates
MEASURMENT OF MICROBIAL ZONE OF INHIBITION
Petridish No. Hole No. Microorganism Antibiotic (ciprofloxacin) concentration* (in microgram / Average Zone of Inhibition in
ml) mm.
ZOI (MM)
200 2.301 26.4 42.2 25
20
100 2.000 22.1 35 15
10
50 1.699 19.9 30.2
5
25 1.398 24.6 0
1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4
12.5 1.097 19.8 LOG[CONC( G/ML)]
DISCUSSION AND CONCLUSION FOR ZOI
The measurement of microbial zone of inhibition experiment was carried out using
teeth sample and Pseudomonas aeruginosa sp.
From the both result and graph it was concluded that micro orgasms from teeth
sample are much more resistant to Ciprofloxacin than Pseudomonas aeruginosa sp.
This clearly indicates that bacterial cell undergo a number of physiological and
phenotypic changes following attachment to a solid surface. These lead to higher cell
resistance to antimicrobial compounds.
These biofilms can induce diseases such as caries, gingivitis and periodontitis which
are among the most common infections of man.
The antibiotic susceptibility is increased many folds when combined with active salt,
this may give a new era to treat different types of teeth disease caused by biofilm
making microorganisms.