1. Technology And Engineering Expertise Interest Chemical analysis

2. Analysis Of Applied Microbiology SMK Negeri 13 Bandung

3. Test Of Contaminant Microbe Total Plate Count ( TPC) Method SMK Negeri 13 Bandung

4. Teknologi dan Rekayasa Learning Objectives  After learning process, the students are expected to be able to:  1. Description the base theory about test of contaminant microbe method Total Plate Count ( TPC).  2. Works admittedly, accurately, aseptic, sterile and having responsibility and care to vicinity area.  3. Do preparation of determination of TPC covers: making media, preparation of practicum equipments, retrieval & preparation of sample.

5. Teknologi dan Rekayasa Learning Objective 4. Do the technical of determination of the germ number method TPC. 5. Do the incubation process of sample TPC. 6. Observes the result of incubation TPC. 7. Do calculation germ number and make conclusions. 8. Report the result of observation

6. Teknologi dan Rekayasa Determination of Cells Number Method and Microbe Colony To determine number of cells and microbe colony can be done with various methods, inter alia : 1. Decants Cup method and diluting serial of which more common is conceived as Total Plate Count method( TPC). 2. Direct microscopic method with apply counting chamber. 3. Cell coloration method in microscopic. 4. Screening method ( Filtration). 5. Turbidity method ( measurement suspension turbidity)

7. Teknologi dan Rekayasa Understanding of Total Plate Count Method (TPC) Total Plate Count is a test of contaminant microbe method with aim to calculate colony total microbe in example of solid and also liquid with cup decants method and serial of diluting

8. Teknologi dan Rekayasa Another Terms From TPC  At some literatures, Total Plate Count method also called as : 1. Determination of Germ Number 2. Determination of Total Plate ( PLT) 3. Total Plate Number ( ALT) 4. Test Contaminant Microbe

9. Teknologi dan Rekayasa Sample Types for TPC Sample Types liquid Sample powder Sample viscid sample Solid Sample congelation Sample  In general all sample types which is in the form of  solid as well as liquid can be counted its number of colonies  microbe ( ALT) use TPC method.  Powder sample types, viscid, solid and congelation  commonly are included into solid sample types.

10. Teknologi dan Rekayasa Solvents, Diluents, Media SolventsDiluentsMedia 1. sterile water 2. Peptone Water 3. Buffered Peptone Water 4. Buffered Distilled Water 5. Phosphate Buffered Distilled Water sterile water 9 mL in test tube 1. Nutrien jell ( SODIUM), to determine total microbe 2. Plate Count Agar ( PCA), to determine total microbe 3. PCA + TTC ( Trifenil Tetrazolium Chlorida) 1%, to determine total bacterium 4. Potato Dextrose jell ( Personal digital assistant) + CHCL3 1%, to determine total fungy

11. Teknologi dan Rekayasa Procedures  Sampling  preparation of Sample  Processing of serial diluting method and cup decants according to schema ( enclosed)  Making of Blanco control media & sterile water  Incubation  Calculation of number of colonies use colony counter  Calculation of Total Plate Number ( ALT)  Description : All processing is done in aseptic

12. Teknologi dan Rekayasa Equipments And Materials Equipments Materials  Sterile Petri dish according to dilution  Graduated pipette sterile 2 ml, according to dilution  Sterile mortar 1 set, for solid sample  Alumunium foil measure 8x8 cm ( sterile) 1 thing, for solid sample  Burner  Incubator  Sterile spatula  Beaker glass 50 ml, 1 thing  Colony Counter  Example of solid food, for example biscuit  Example of beverage  Sterile water 9 ml in test tube according to dilution  PCA 15 ml in test tube according to dilution.  Sterile Water or solvent appropriate for solid sample type

13. Teknologi dan Rekayasa Job Schema of TPC Liquid Sample

14. Teknologi dan Rekayasa Job Schema of TPC Solid Sample  After obtaining dilution of 10-1 continuing the process of liquid sample as job schema. Number of dilutions done depends on to level of sample contaminant. For sample with level of high contaminant, dilution done from starting low dilution until dilution of 10-7, while for sample with level of low contaminant, maximum dilution is 10-4 or 10-5.

15. Teknologi dan Rekayasa More and Less of TPC Method  This calculation method is the most sensitive way to count microbe, because the reasons as follows : 1. Only cell that is above the ground calculated. 2. Some types of microbe can be counted at the same time. 3. Applicable to isolate and identification of microbe because colony formed comes out of one microbe cells with specific growth vision.

16. Teknologi dan Rekayasa More And Less Of TPC Method  Less of TPC method:  1. Result of calculation doesn't show the number of microbe cells actually, because some nearby cells possibly forms one colony.  2. different Medium and condition possibly yields different value.  3. Microbe grown must be able grow at solid medium and forms compact colony and clear, which disseminates.  4. Requires a few days for preparation and incubation time, so that growth of countable colony.

17. Teknologi dan Rekayasa Calculation Of ALT  calculate colony microbe at each cup use colony counter.  If there are two or more cups with level of successive dilution contained 30-300 colonies, hence :  If amounts of colonies at higher dilution ( e.g., 10-3) 2 times bigger than amounts of colonies at dilution under it ( e.g., 10-2), hence total plate numbers determined by level of lower dilution ( 10-2).  If amounts of colonies at higher dilution ( e.g., 10-3) 2 times smaller than amounts of colonies at dilution under it ( e.g., 10-2), hence total plate number determined by both level of the dilution that is 10-2 and 10-3.

18. Teknologi dan Rekayasa Calculation Of ALT  If there is not one of colony which grow at Petri dish, hence ALT expressed : smaller than ( 1) one x factor low dilution; example:  Low dilution is null ( P=100) ALT<1  Low dilution is 10-1 ( P=10-1) ALT<10  Total Plate Number ( ALT) only written in two numbers, the next is rounded up :  If > 5 is rounded up upward  If < 5 is rounded up below

19. Teknologi dan Rekayasa Calculation of ALT  The explanation of calculation of ALT in short can be seen at the table below: ExamplesDilutionsAmount of Colonies ALT a.P=10 -2 P=10 -3 140.10 2 (=14.10 3 ) 32.10 3 32.10 3 > 2x14.10 3 hence ALT = 14.10 3 colony/gram b.P=10 -2 P=10 -3 293.10 2 (=29,3.10 3 ) 41.10 3 41.10 3 < 2x29,3.10 3 (29,3+41/2) (10 3 ) hence ALT = 35,12.10 3 c.P=10 0 (lowest) P=10 -1 (lowest) 0000 ALT < 1 ALT < 10 d.If ALT = 35,12.10 3, written by; ALT = 35.10 3 If ALT = 35,62.10 3, written by; ALT = 36.10 3

20. Teknologi dan Rekayasa Excercises  Count ALT from the data as follows! Dilution10 0 10 -1 10 -2 10 -3 10 -4 10 -5 Amount of Colony>300 150502510  Answer : a. Data entering condition calculation is: 10 -2 = 150 & 10 -3 = 50 b. Amount of colony is multiplied by dilution factor: 150 X 1/10 -2 = 150.10 -2 = 15.10 -3 50 X 1/10 -3 = 50.10 -3 c.50.10 -3 > 2 X 15.10 -3 d.Hence ALT = 15.10 -3 colony/ml

21. Teknologi dan Rekayasa Picture Of Colony Counter

22. Teknologi dan Rekayasa