Full transcript of “Pengujian Antimikroba/Antibakteri/Kirby Bauer/Paper Disc/Uji Daya Hambat/Mc Farland/Turbidimetri”. Click a timestamp to open the video at that moment. Need another video? Use the YouTube transcript generator.
0:00 Hello [Music] Assalamualaikum friends Today we will conduct antimicrobial testing using the Kirby-Bauer method as usual before starting the test we first clean our work table using 70 percent alcohol in this test we use Muller hinten agar or MH medium which has been sterile then we also need a
0:45 sterile petri dish Hi and a bunsen burner Hi then before that we first clean the outside of the medium bottle using 70% alcohol pour the medium aseptically by placing the mouth of the bottle over the bunsen burner then we pour the medium into a sterile petri dish like this we pour the main in Um with a depth of approximately five MM Hi pour the medium into all the petri dishes hi hi Hi
1:34 then we let it sit until the medium pauses Hi while waiting for the medium Now we will make a fresh spice extract here there is fresh ginger and garlic that we have peeled and washed using boiled water then we drain it here there is also a mortar and pestle that we sterilize in the oven cut the ginger and garlic like this to make it easier in the grinding process
2:20 Hi prepare a mortar and Then we puree the garlic and ginger, first we will puree the garlic, we will crush it near the Bunsen burner until it is smooth like this, now we will puree the ginger, after the garlic is smooth, then we add 1 sterile Wade, approximately 10 milliliters,
3:14 after that we homogenize it and grow it again, the fresh extract is transferred into a small petri dish, then we label the ginger, the same, after it is smooth, then we add 1 like this, hehehehe, now we will make a suspension of test bacteria, where the test bacteria used are pathogenic bacteria, Escherichia coli and Staphylococcus aureus,
4:00 now this is the test bacteria that we have incubated for approximately one day, its regeneration can be seen in the previous video, yes, the characteristics of bacterial growth on slant agar are the formation of a layer of yellowish-white cream on the surface of the agar, so that we will make a bacterial suspension by dissolving it with a physiological NaCl solution of 0.85 percent and comparing it visually with a standard McFarland solution of 0.5. These tubes contain a solution of NaCl 0.85 percent which we will use for
4:45 dissolve bacteria and this is a mcfarlane solution, the turbidity of the suspension will be measured using a spectrophotometer with a wavelength of 600 nanometers and an absorbance of 0.1, an absorbance value of 0.1 is equivalent to 1.5 * 10 ^ 8 c [ __ ] per milli Now let's pay attention to the technique of making a bacterial suspension with a standard mcfarland solution of 0.5, a physiological 0.85 percent NaCl solution
5:34 will be poured into the slanted agar Now let's pay attention to the technique that Indonesia does pour the NaCl solution by doing it over a bunsen flame, then we pour the solution approximately half the height of the slanted agar, then don't forget to close it using a bokep stopper, then we stir or homogenize it
6:20 slowly, be careful not to destroy the agar, we can see the bacteria that grow on the surface of the slanted agar, now it has been dissolved by the NaCl solution, now we see the color becomes more cloudy, friends Hi, next, take a new NaCl tube, then pour the suspension into the tube that was in the slanted agar, we poured a few drops,
7:10 then we homogenize the agar, then compare the turbidity with the naked eye with the tube reaction containing 0.5 mcfarland standard solution now we compare its turbidity with NaCl solution after adding bacterial suspension a few drops of NaCl solution become cloudy Now we compare it with Mcfarlane standard solution If it is more cloudy than the standard then we have to add NaCl solution
7:56 If it is more faded then add more suspension solution that is on the slant agar maybe if we see the ecoli suspension that we made is more cloudy compared to Mcfarland then we have to add NaCl solution pour NaCl a few drops until the turbidity = Mcfarland 0.5 A mogen kan again then compare it with the standard solution
8:45 Hi if we see the suspension that we made the turbidity is close to the turbidity of Mcfarlane then we label it Ice Okay so that it is not mixed up then we will make a suspension of Staphylococcus aureus bacteria the method is the same as before yes friends we pour NaCl until half of the test tube from the slant agar then
9:30 we homogenize it slowly so that the test bacteria Dissolve by the NaCl solution no sir a few drops of suspension on the slant agar into the test tube containing the new NaCl then homogenize it back to Yes, I said the same as earlier we compared it with the standard solution visually hello hello Launcher then we see the
10:18 suspension solution that was made is more turbid compared to mcfarlane then we add physiological NaCl solution After that we homogenize and compare it again if it feels the same then give a label then we will check the absorbance using a spectrophotometer this is the spectrophotometer instrument that we will use first we set the wavelength at 666 m then click the 0 ABS button
11:04 then the display will show the Blank setting and absorbance 0.000 a.in is the suspension of the bacterium solution that we made earlier and we will measure its absorbance using a spectrophotometer this is a cuvette the suspension will be poured into the cuvette to then measure its absorbance with before we measure the test bacterial suspension then the first thing we measure is the mcfarland solution as a blank we pour the blank solution into the cuvette up to the
11:50 specified limit then we put it in the spectrophotometer we close it and we click the no Labs button then the Blend setting appears and 0.0000 now we pour the ecoli bacteria suspension into the cuvette then we measure absorbance Hi according to the literature the absorbance reaches a value of 0.1 so it is equivalent to mcfarland 0.5 Sir bacterial suspension into the cuvette up to the cuvette boundary mark
12:43 insert the cuvette into the spectro then we see the absorbance read on the display adjust the measured aids It turns out 0.01 one while the appropriate Standard is absorbance 0.1 then we have to add a concentrated suspension so that it tilts earlier so that the absorbance increases after and sih is added homogenized Then pour the suspension back into the cuvette After that we measure it in the spectrophotometer read the a value on this display here we
13:31 read the a value of 0.1 15 record it in the log book because the absorbance has been achieved or in accordance with the standard then we measure for the Staphylococcus aureus bacterial suspension the method is the same as before yes after being measured using a spectrophotometer it can be seen on the display the a value obtained for Staphylococcus aureus is 0.1 08 because it is in accordance with the standard then we just have to record it in the log now we have entered At the antimicrobial testing stage we see the MH medium that we poured earlier is already there
14:19 now we label the icon P3 using a permanent marker with the code SC for eserciacoli and Esa for Staphylococcus aureus we write on the top of the petri dish like this Indonesia then Strike each dish with the suspension that we have measured earlier with the absorbance that we have Cakra trick using Katon sweet sterile pay
15:07 attention to how to open the tube stopper using your finger beginning then do it on the bunsen dip the bribe up and down then squeeze it against the wall of the tube OK Google take a petri dish containing MH agar then we Strike with the direct or zigzag method pay attention to how to
15:53 open the petri dish do it in the fire then we Strike in a vertical direction from top to bottom zigzag like this close the dish then we turn it then we Strike vertically again like this finally we Strike the edge of the petri dish Hi for the next answer dip the swap back into the bacterial suspension then Strike with the technique as before the
16:40 swap with the same bacteria does not need to be replaced yes if there is a change of bacteria then the swap must be replaced with a new one Indonesian film if it is finished nickname bribe into the tube Hi break some then close the tube using the tube stopper should not be
17:27 washed immediately yes must be described first first because it contains pathogenic bacteria let the cup stand for about five minutes in addition to the fresh ginger and garlic extracts that we have prepared earlier we also use positive controls, namely antibiotics and negative controls, namely sterile distilled water, this is a round disk type with a diameter of 6 mm that we have sterilized pepper here later we will soak it in the four solutions above this is sterile tweezers used to take paper discs 1 tweezers are used for one solution or sample
18:14 that Hi the next second we label the bottom of the cup with a permanent marker with the code Kak positive for positive control Kak negative for negative control J for ginger and BP for garlic take the paper disk using sterile tweezers then insert it into the cup containing the sample solution or extract for about 10 minutes only
19:03 Hi take that this that has been soaked using tweezers put it on the lid of the cup then tap it so that the extract is j & t Hi Place the paper according to the code or label give space between samples press the paper disk slowly so that it sticks to the surface of the cup do the same thing Place the sample according to the code that we have written [Music]
20:22 Hi this [Music] after all is done then we wrap the petri dish using cling wrap like this finally we put in the incubator we incubate at 37° Celsius for 24
21:09 hours [Music] Yes [Music] indeed [Music] The following are the results of positive and negative controls we can see there is a clear area on the surface so that this means that there is inhibition of the test bacteria by antibiotics the inhibition zone is a clear area
21:56 around the paper disk which can indicate that there is activity of the test bacteria the diameter of the clear zone of antibiotics against Staphylococcus aureus here we see very wide the diameter of the clear zone of antibiotics against escherichia colli bacteria is smaller compared to the size us inhibition zone now we see the diameter of the clear zone of fresh garlic extract if we look at the clear zone formed in both test bacteria it is
22:41 almost the same The following are the results of observations of ginger extract we can see the clear area for Staphylococcus aureus is larger than for escherichia colli bacteria use a ruler to measure the diameter of the clear zone How to measure it we see in the picture yes here one cup has two clear zones this is simple and duplicate data yes pay attention to one of the clear zones for the test bacteria Staphylococcus aureus here there are two yellow lines of clear zones calculated vertically and horizontally measure the area of the clear Dona vertically from top
23:29 to bottom including the paper disk then calculate the horizontal line the formula for calculating the inhibition zone is the diameter of the vertical clear zone minus the diameter of the paper disk plus the diameter of the horizontal clear zone minus the diameter of the paper disk divided by two the inhibition zone of ginger extract against saw Reus 29 minus 6 diameter perdif Then add 30 minus 6 divided by two equals 25 mm diameter of the inhibition zone for fresh ginger extract The following are the measurement results of the vertical and horizontal clear zones
24:17 for fresh ginger garlic extract and positive controls please calculate how much the inhibition zone is according to the previous formula After you calculate the inhibition zone from the previous table now compare the observation results with the standard table Determine the antimicrobial effectiveness of fresh garlic and ginger extract whether it is a compound that is resistant intermediate or sensitive [Music] [Applause]
25:03 [Music]
