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Technology And Engineering
Expertise Interest Chemical analysis
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Analysis Of Applied Microbiology
SMK Negeri 13 Bandung
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TESY OF ANTIBIOTIC POTENTIAL
SMK Negeri 13 Bandung
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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. Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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Understanding of Antimicrobial
Antimicrobial is the character of material which can pursue or kills microorganism growth. Teknologi dan Rekayasa
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Types of Antimicrobial
Natural Materials Chemical Materials or Compounds Antibiotics Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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 Teknologi dan Rekayasa
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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). Teknologi dan Rekayasa
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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 Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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. Teknologi dan Rekayasa
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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 Teknologi dan Rekayasa
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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 Teknologi dan Rekayasa
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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 Teknologi dan Rekayasa
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Example of Calculation of RPa
RPa sample= (0,88 + 0,89 + 0,87)/3 = 0,88 Conclusion: RPa < 1, hence, bad quality antibiotic Teknologi dan Rekayasa
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Example Of Observation of Resistance Zone
Teknologi dan Rekayasa
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Factors Influencing The Result Of Antibiotic Potential Test
Methods are used Materials and equipments Microbe Test is used Teknologi dan Rekayasa
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terima kasih Teknologi dan Rekayasa
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