The Effectiveness Test of Bangun-Bangun Leaves Extract (Plecthranthus Ambonicus) Against Streptococcus Pyogenes and Salmonella Typhi Bacteria

Background Bangun-bangun leaves (Plectranthus amboinicus) are part of the Lamiaceae family. (1) (1) Bangun-bangun leaves have a characteristic aromatic leaves aroma used as a medicinal plant (2). According to research conducted by (Dalimunthe et al., 2016) to find the use of this plant as an antibacterial because it contains compounds such as flavonoids, tannins, tripenoids, saponins, and essential oils. On the bangun-bangun leaves there is carvacrol, which is an essential component that has antibacterial activity in positive or negative gram bacteria. streptococcus pyogenes bacteria are vectors of suppurative infectious diseases and other diseases such as pharyngitis. While salmonella typhi bacteria is a vector of the disease which is usually called typhus and the medical language is typhoid fever. The purpose of this study was to determine that the bacteria salmonella typhi and streptococcus pyogenens can be inhibited by the leaves extract. The benefit of this research is to increase public understanding and knowledge in the use of bangun-bangun leaves. The method used in this study is an experimental method and post test only controlled group design. For the concentration used, namely 5%, 10%, 15%, 20%, 25% and this strength test is done by inserting the extract into a petri dish that has a bacterial culture. Data processing uses one-way ANOVA with the aim of comparing each variable. To see the extract against bacteria. The results showed that there were variations in the diameter of the inhibition zone formed by the concentration of the extract. And the most effective concentration to inhibit streptococcus pyogenes bacteria was a concentration of 25%, the inhibition zone diameter was 19.5mm, while salmonella typhi bacteria had a concentration of 25% and the inhibition zone diameter was 37.7m.


Introduction
Streptococcus β hemolyticus group A or can be called streptococcus pyognes which is included in coccus-shaped bacteria with a chain-like and nonmotile shape and has a diameter of 0.6 -1.0 micrometers and is facultative anerobic, and includes (3) . Metabolism of streptococcus which is fermentative and requires media that usually uses blood agar media but can also be used MHA media, these bacteria can live well at optimum temperatures, namely temperatures of 370c and ph of 7.4-7.6 and including bacteria have capsules containing hyaluronic acid.
In general, bacteria can infect humans that can cause suppurative diseases such as pharyngitis, impetigo, erysipelas, and scarlet fever, there were 9000-11500 cases of streptococcus pyogenes and the mortality rate from these cases was 1000-1800 deaths from all cases United States of America There is a case of streptococcus pyogenes which is a very important case in Indonesia for the treatment used is a ampicillin but there are many resistant cases during treatment and there are side effects that trigger other cases so alternative treatment is needed to

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Journal Homepage: https://www.jurnalkedokteranunsri.id/index.php/BJI/index minimize side effects and reduce resistance rates when treatment is carried out. (4).
Salmonella tyhi is a disease vector of typhoid fever or commonly referred to as typhoid, salmonella typhi is included in the entererobacteriaceae family which is rod-shaped and belongs to the gram-negative group and has flagella which are facultative anaerobes. As well as these bacteria have varying levels of invasion and can usually infect humans from minor infections, for example, the disease caused is diarrhea and severe infections which are usually caused by typhoid fever (5) .
Typhoid fever or can be called typhus abdominalis which is an acute disease that infects the small intestine but can also occur in the bloodstream caused by salmonella typhi. Clinical symptoms that often occur when infected with these bacteria are high fever, abdominal pain, red and red spots and weakness, lethargy and many other clinical symptoms that arise when infected by these bacteria. Usually the spread of bacteria or the affected person because it is contaminated through food or drink which causes clinical symptoms of acute typhoid fever infection. And there are several factors that influence the incidence of typhoid fever, namely the lack of cleanliness by an individual, a dense home or living environment and also insufficient water availability and a decline in the system. immunity or a person's immunity.
In medicinal plants, the bangun-bangun leaves (plectranthus ambonicus) belong to the lamiaceae family. woke leaves is a traditional plant that has a distinctive aroma and is found in tropical areas and is known as an aromatic plant which can also be useful as a traditional medicine. The bangun-bangun leaves of these shapes are found in many Asian and African countries around 80%. The bangun-bangun leaves of these shapes are usually used to increase breast milk during breastfeeding but apart from that, they can be consumed as food ingredients and can also be used as antibacterials which can inhibit the growth of bacteria or kill bacteria that cause infection.

Research Methods
This study used the post test only controlled group design research method as its design plan. where this design is used because the sample was selected without any pretest treatment first.

Location and time
The location used in the research was in the Biomolecular Laboratory at the Medical Faculty of the Prima Indonesia University on the path of pot No.1 Ayahanda, Medan Utara, which was chosen as a good and ideal place for the extraction process in September 2020 which was then carried out by testing the extract as antibacterial from streptococcus pyogenens and salmonella bacteria. Typhi, which is located in the Biomolecular Laboratory of Prima Indonesia University in September 2020.

Research Samples
In this study we used the research sample that was used at the time of the study was the bangun-bangun leaves (plencatrathus ambonicus) which came from the wake-up plant found in the Lubuk Pakam area, North Sumatra.

Method of collecting data
Data obtained from Bangun-bangun leaves extracts (plecthranthus ambonicus) obtained during research conducted in the laboratory, which used primary data during the test of the effectiveness of the leaves extracts in inhibiting streptococcus pyogenes and salmonella typhi bacteria.

Research Instruments
During the research, the equipment indispensable

Extraction Method
On the bangun-bangun leaves of the shapes that were given the extract treatment of this research using the maceration method. For the solvent to be used in this study is 96% ethanol as a solvent. The bangunbangun leaves are collected as much as 10 kg after drying into a drying place to be 5 kg then grinding using a blender as much as 600 grams then the maceration process is carried out using 96% ethanol solvent which is needed as much as 600 ml mixed with and left to stand for 72 hours at room temperature.
The solution is filtered and then a rotary evaporator is used to evaporate the solvent. Then the extract is placed in a water bath until the extract is thick and good for use, then proceed by dividing into 5 concentrations, namely: 1. 5% concentration: the concentration of 5% bangunbangun leaves extract is taken as much as 10 ml mixed with 100 ml of aquabidest.
2. 10% concentration: The 10% concentration of 10% of the bangun-bangun leaves extract is taken as much as 10ml mixed with 100ml of aquabidest.
3. Concentration of bangun-bangun leaves extract 15%: The concentration of 15% of bangun-bangun leaves extract is taken as much as 10 ml mixed with 100 ml of aquabidest.
4. 20% concentration: The concentration of 20% of the bangun-bangun leaves extract is taken as much as 10 ml mixed with 100 ml of aquabidest.
5. 25% concentration: The concentration of 25% of bangun-bangun leaves extract is taken as much as 10 ml mixed with 100 ml of aquabidest.

Test the effectiveness of the bangun-bangun leaves extract
The Biomolecular Laboratory of Prima Indonesia University was chosen as the place to carry out the effectiveness test of the leaves of the shapes. The test of the effectiveness of the leaves extract as antibacterial was carried out in vitro which will be used for the samples in this research.
1. In supensi bacteria streptococcus pyogenens and salmonella typhi which has been smeared.

2.
A cotton stick is used to apply to MHA media.
3. Then the disc paper that has been provided that has been dripped by the leaves extract concentration rises for 15 minutes.
4. Next, it will be wrapped in plastic wrap and followed by wrapping it with aluminum foil 5. Then incubate in an incubator with a temperature of 37 0 C for 24 hours.
6. Furthermore, the incubation is complete, followed by measuring the diameter of the inhibition zone

Data analysis
After obtaining the inhibition zone diameter data from the antibacterial test of the bangun-bangun leaves extracts of the streptococus pyogenens and salmonella typhi bacteria, then processing the data to be tested through the one way ANOVA test via SPSS software.

Results
The research was carried out at the Prima Indonesia Biomolecular Laboratory on July 11-20 October 2020.

Amboinicus) Against Growth of Streptococcus pyogenens Bacteria.
In the bangun-bangun leaves extract (plectharnthus ambonicus) on the growth of the Streptococcus pyogenens bacteria, which in this study used the disc diffusion method, it was shown that the inhibition zone was measured using a caliper. The results of the study used a concentration of 5%, 10%, 15%, 15%, 20%, 25%, the inhibition zones were obtained which are listed in the following figure and figure: In this study we will perform a statistical test using the one way ANOVA test, but provided that the data for each variable in the study must be normally distributed

Research Advantages
In this study we can find out the benefits of the bangun-bangun leaves extract in terms of inhibiting the growth of the Streptococcus pyogenes bacteria, which are gram-positive bacteria and salmonella typhi which are gram-negative bacteria. and in this study using equipment that is relatively simple and easy to do.

Research Weaknesses
In this study, we obtained the results that the bangun-bangun leaves extract was able to inhibit the growth of streptococcus and salmonella typhi bacteria.
However, the inhibition zone diameter did not exceed the zone diameter from the positive control and the maceration process took a long time to work.