1. Technology And Engineering
Expertise Interest
Chemical analysis

2. Analysis Of Applied Microbiology
SMK Negeri 13 Bandung

3. TESY OF ANTIBIOTIC POTENTIAL
SMK Negeri 13 Bandung

4. LEARNING OBJECTIVES
After learning process, the students are expected to be able to:
1. Description the basic theory of water analysis and food in MPN Method.
2. Works admittedly, accurately, aseptic, sterile and having responsibility and care to vicinity area.
3. Do preparations of Antibiotic Potential test, covers of making media and preparation of sterile equipments.
4. Preparation of microbe suspension test.
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5. Learning Objectives 5. Do Antibiotic Potential Test.
6. Observe, make a note, and calculation of antibiotic potential test.
7. Report the result of observation and calculation of antibiotic potential test.
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6. Understanding of Antimicrobial
Antimicrobial is the character of material which can pursue or kills microorganism growth.
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7. Types of Antimicrobial
Natural Materials
Chemical Materials or Compounds
Antibiotics
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8. Understanding of Antibiotic
Antibiotic is a matter yielded by a microorganism or its generation which at low concentration can pursue or annihilates growth of other microorganism.
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9. Mechanism of Antibiotic Activity
Cell wall breakdown
Changes cell permeability
Changes protein molecule and nucleic acid
Pursues enzyme activity
Pursues nucleic acid synthesis and protein
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10. Clacification of Antibiotics
Based on the antibiotic activities, it is grouped into two, that is :
1. Broad spectrum antibiotic ( antibiotic is earning pursues or kills growth of Gram-positive bacteria and Gram negativity).
2. narrow spectrum antibiotic ( antibiotic which only can pursue or kills one of group of bacterium Gram only).
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11. Factors which Influencing Antibiotic Energy Activity
Density of microorganism population
Sensitivity to antibiotic material
Material volume sterilized
The duration antibiotic material application at microorganism
Concentration of antibiotic material
Temperature
Organic material content
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12. Understanding Of Antibiotic Potential Test
Antibiotic potential test is a series of works with aim to know ability an antibiotic material in pursuing or kills a microorganism.
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13. Method of Antibiotic Potential Test
Determination method with cylinder plate ( Jelly Diffusion method)
This method based on diffusion antibiotic from cylinder attached by perpendicular at jelly solid layer in Petri dish or plate, so that microbe added pursued its growth at circle area or zone around cylinder which is containing antibiotic solution.
Turbidity method
This method based on growth resistance of microbe breeding in equal antibiotic solution, in liquid medium which can grow microbe swiftly if there are no antibiotics.
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14. Procedures of Antibiotic Potential Test
Prepare test microbe suspension.
Prepare primer standard solution by the way: weight 25gram antibiotic standard then dissolves in volumetric flask 250 mL with solvent LDF2 ( quantitatively and aseptic).
Create various concentration of standard solutions( S1, S2, S3 ) by the way :
S1 : enter 5 mL primer standard solution into volumetric flask 100 mL then dilute by using sterile water.
S2 : enter 4 mL primer standard solution into volumetric flask 100 mL then dilute by using sterile water.
S3 : enter 3 mL primer standard solution into volumetric flask 100 mL then dilute by using sterile water.
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15. Procedures of Antibiotic Potential Test
Prepares condensation of mains sample by the way : deliberates equivalent antibiotic sample with 25 active matter grams then dissolves in volumetric flask 250 mL with solvent LDF2 ( quantitatively and aseptis).
Create various concentration of sample (C1, C2, C3) by the way :
C1 : enters 5 mL prime sample into volumetric flask 100 mL then dilute by using sterile water.
C2 : enters 4 mL prime sample into volumetric flask 100 mL then dilute by using sterile water.
C3 : enters 3 mL prime sample into volumetric flask 100 mL then dilute by using sterile water.
Prepares 2 sterile Petri dishes and give pattern according to picture under.
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16. Procedures of Antibiotic Potential Test
Enters 1mL bacterium suspension test into 100 mL medium antibiotik2 ( AB2) then homogenizes.
Enters ± 15 mL bacterium mixture with AB2 into each Petri dish which has been given pattern, smooths down and lets frost.
Well create at each pattern by using media punch.
Enters ± 2 drip solutions S1,S2,S3,C1,C2,C3 into each well as according to pattern.
Second incubation of Petri dish at temperature 370C during 48 hours
Measure Resistance zona ( transparent area around antibiotic solution at Petri dish) which is formed.
Calculate Ratio Potensi Antibiotik from sample tested.
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17. Storage Pattern of solution at Well
Descriptions :
S1 = Standard 1
S2 = Standard 2
S3 = Standard 3
C1 = Example 1
C2 = Example 2
C3 = Example 3
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18. Calculation of Antibiotic Potential Ratio
RPa1 = Φ average C1/ Φ average S1
RPa2 = Φ average C2/ Φ average S2
RPa3 = Φ average C3/ Φ average S3
Descriptions :
RPa > 1, hence good quality antibiotic
RPa < 1, hence
Bad quality antibiotic
RPa = 1, hence
quality of proportional antibiotic with standard
Φ = resistance zone diameter
RPa Sample :
(RPa1 + RPa2 + RPa3)/3
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19. Example Of Calculation Of
Observation Data Of Incubation Result No
Variation Tested
Diameter Resistance Zone (Φ) (Cm)
Petri 1
Petri 2
averages 1. S1
1,20
1,40
1,30 2. S2
1,50 3. S3
1,60
1,70
1,65 4. C1
1,10
1,15 5. C2
1,25 6. C3
1,45
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20. Example of Calculation of RPa
RPa sample= (0,88 + 0,89 + 0,87)/3
= 0,88
Conclusion: RPa < 1, hence, bad quality antibiotic
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21. Example Of Observation of Resistance Zone
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22. Factors Influencing The Result Of Antibiotic Potential Test
Methods are used
Materials and equipments
Microbe Test is used
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23. terima kasih
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