Uji Koefisien Fenol dan Jumlah Koloni Bakteri Uji Pada Perlakuan Kombinasi Infus Daun Sirih-Kemangi

Summary

This transcript outlines a detailed experimental procedure for testing the antibacterial effects of **betel leaf** (sirih) and **kemangi** (SpongeBob leaf) infusions. The process includes preparation of infusions, nutrient broth, and phenol coefficient testing with different bacterial strains, such as *Staphylococcus aureus*, *E. coli*, *Salmonella typhi*, and *Pseudomonas aeruginosa*. The experiment tests the antibacterial activity over varying time intervals (5, 10, and 15 minutes) and dilution concentrations. The procedure concludes with colony count testing and a thorough cleaning process. The study aims to explore natural remedies' potential in inhibiting bacterial growth.

Takeaways

• 😀 The research begins with the researcher washing their hands and preparing the materials, including Spongebob hand gloves and fresh basil and betel leaves.
• 😀 After washing and cutting the basil and betel leaves, the researcher weighs them at 100 grams before infusing them in a pot with 100 mL of aquades.
• 😀 The leaves are heated for 15 minutes at 90°C to make an infusion, after which the infusion is filtered and separated from the solid materials.
• 😀 The basil infusion process is repeated as described, and the infusion is stored in a laminar flow hood after filtering to maintain sterility.
• 😀 Nutrient broth (NB) is prepared by dissolving 13 grams in 1 liter of aquades and sterilized in an autoclave for 15 minutes at 121°C under 2 ATM pressure.
• 😀 The next step involves preparing test tubes for phenol coefficient testing by adding 1 mL of the basil and betel leaf infusions into separate tubes, which are then diluted with aquades at various concentrations.
• 😀 After dilution, bacterial cultures are introduced into the tubes, and the bacteria are incubated under controlled conditions to assess the effectiveness of the infusions.
• 😀 After 5, 10, and 15 minutes of contact, the tubes are sampled for bacterial growth, and bacterial contamination is tested for four different bacteria: Staphylococcus aureus, Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa.
• 😀 In a separate test, the researcher prepares nutrient agar and incubates the bacterial samples at 37°C for 24 hours to count colony growth and assess bacterial inhibition.
• 😀 All used equipment, including test tubes and petri dishes, is washed and sterilized to ensure no contamination occurs during the testing process.

Q & A

What is the purpose of preparing basil and betel leaf infusions in this experiment?

The purpose of preparing the basil and betel leaf infusions is to test their antibacterial properties against various bacterial strains, including *Staphylococcus aureus*, *Escherichia coli*, *Salmonella typhi*, and *Pseudomonas aeruginosa*.

What steps are involved in preparing the basil and betel leaf infusions?

The steps include washing and slicing the leaves, weighing 100 grams of each, adding them to 100 mL of distilled water, heating the mixture to 90°C for 15 minutes, filtering the infusion to separate the liquid from the leaves, and then cooling it before further use.

Why is the nutrient broth sterilized in the procedure?

The nutrient broth is sterilized to prevent contamination, ensuring that the test results accurately reflect the antibacterial effects of the infusions and not the presence of other microorganisms.

How is the phenol coefficient test conducted in this experiment?

The phenol coefficient test is conducted by adding 1 mL of each infusion (basil and betel) to dilution tubes, diluting them with aquades, then adding nutrient broth and introducing a bacterial sample. The samples are incubated for 24 hours at 37°C, and the bacterial growth is observed after contact times of 5, 10, and 15 minutes.

What does the phenol coefficient test measure?

The phenol coefficient test measures the antibacterial effectiveness of the infusions by comparing the reduction in bacterial growth at different dilutions and contact times.

What is the significance of using different contact times (5, 10, and 15 minutes) in the experiment?

Using different contact times helps assess how the antibacterial efficacy of the infusions changes over time, determining the optimal duration needed for effective bacterial inhibition.

Why are multiple bacterial strains used in the experiment?

Multiple bacterial strains are used to test the broad-spectrum antibacterial potential of the basil and betel leaf infusions, evaluating their effectiveness against a variety of harmful bacteria.

How is the colony count test performed?

In the colony count test, nutrient agar is prepared and sterilized. The infusions are added to Petri dishes with bacterial samples, followed by the addition of nutrient agar. The dishes are then incubated for 24 hours, and bacterial colonies are counted to determine the effectiveness of the infusions in inhibiting bacterial growth.

What is the purpose of autoclaving the nutrient broth and nutrient agar?

Autoclaving the nutrient broth and nutrient agar sterilizes them, ensuring no microbial contamination occurs, which is essential for accurate testing of the infusions' antibacterial properties.

What role does the Bunsen burner play in the procedure?

The Bunsen burner is used for sterilizing tools, such as the inoculating loop (Ose), before and after transferring bacterial samples to prevent cross-contamination and ensure accurate results.