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Antibacterial Bioactivity from Extract of Reundeu Caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
Noverita1 and Ernawati Sinaga 1 Faculty of Biology.
Universitas Nasional.
Jakarta Corresponding author: ernawatisinaga@unas.
Abstract Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
are two examples of plants commonly used by rural tribal communities as medicinal ingredients, one of which is by the Baduy tribal community.
These two plans are usually used by local people to treat infectious diseases .
and stomach aches.
Therefore, it is necessary to prove the medical potential of these plants.
This study was conducted with the aim of knowing the antibacterial potential of the extracts of the Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
plant extracts against the growth of Escherichia coli.
Salmonella typhi.
Staphylococcus aureus, and Bacillus cereus.
The method used in this study is the Disc Diffusion method.
The results showed that Reundeu caret and Honje plant extracts were only able to inhibit the growth of Staphylococcus aureus and Bacillus cereus.
The antibacterial activity of the plant extracts of Rendeu Caret and Honje was moderate to strong.
This study concluded that Honje and Reundeu caret plant extracts were more potential to be used to treat infectious diseases .
Keywords: Antibacterial.
Honje (Etlingera hemisphaeric.
Reundeu caret (Staurogyne zone of inhibition INTRODUCTION Indonesia is a country that has the largest population in the world, so many problems arise in society, one of which is health.
Health is one of the problems that are closely related to medicinal ingredients.
We can find various types of drugs in hospitals, pharmacies, drug stores, and even stalls.
The sources of the basic ingredients for making these drugs are very diverse, some are made from natural chemicals and some are made from synthetic chemicals.
The use of drugs derived from synthetic chemicals raises problems because these materials when used in excess will be harmful to the wearer.
One of the many efforts currently being made is to look for natural medicinal ingredients sourced from medicinal plants.
The use of plants as medicinal ingredients is usually done by people who live in rural areas, especially by traditional people who live far from urban areas and are very tied to the culture of their community, such as the Baduy tribe who live in Kanekes village.
Leuwidamar district.
Lebak regency.
Reundeu caret (Staurogyne longat.
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Honje (Etlingera hemisphaeric.
are two examples of plants commonly used by the Baduy people for food and medicine.
According to Mariani .
Reundeu caret is a plant that is commonly found in the mountains in the tropics.
For the Sundanese people, these plants are used as fresh vegetables, besides that these plants are also used as medicinal ingredients.
Honje (Etlingera hemisphaeric.
or better known as kecomrang is a group of spice plants from the Zingiberaceae .
inger-ginge.
These plants have sour flowers and fruit with a distinctive fragrant smell, are mixed ingredients, and at the same time flavoring spices for various dishes in Indonesia.
Besides, this plant is also efficacious as a medicinal ingredient, which is used as a drug for skin diseases, cancer, tumors.
The potential of these plants as medicinal ingredients is related to the content of their bioactive compounds.
According to Mariani .
Reudeu caret contains bioactive compounds in the form of flavonoids, saponins, steroids/triterpenoids, tannins, and phenols.
Meanwhile, according to Jackie et al.
Honje (Etlingera hemisphaeric.
contains bioactive compounds in the form of glycosides, polyphenols, and flavonoids.
The treatment method used by the community is very simple, namely by pounding this material until smooth, then smeared on the sick part, or by brewing it with hot water and drinking it.
To prove the efficacy of plants as medicinal ingredients, especially drugs for infection by microorganisms .
acteria, fungi, and viruse.
, it is necessary to prove medically through antimicrobial tests, one of which is antibacterial.
The test bacteria commonly used in research are bacteria that are commonly found in everyday life, and have a fairly high infection rate, and are opportunistic to other organisms.
These bacteria can be from groups of Gram-positive bacteria such as Staphylococcus aureus and Bacillus cereus which produce thermostable toxins, and can also come from groups of Gram-negative bacteria such as Escherichia coli and Salmonella typhi that cause infections in the digestive tract (Adams and Moss, 2.
Seeing the potential that exists in the two plants (Reundeu caret and Honj.
, and there is still little information regarding their antibacterial potential, this study was conducted to know the antibacterial power of the plant extracts of Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
against bacterial growth Escherichia coli.
Salmonella typhi.
Bacillus cereus, and Staphylococcus aureus.
The hypotheses to be tested in this study are:
Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
extracts have antibacterial activity against the test bacteria used.
The test bacteria had different responses to the Reundeu caret and Honje plant
METHOD
Place A sampling of medicinal plants Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
, was carried out in the village of the Baduy Dalam tribe.
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Kanekes Village.
Lebak Banten Regency, and the antibacterial activity testing was carried out at the Microbiology and Genetics Laboratory of the National University Integrated Lab.
Bambu Kuning.
Pasar Minggu South Jakarta.
Materials and Tools The materials used in this study include extracts of medicinal plants Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
, isolates of Staphylococcus aureus.
Bacillus cereus.
Staphylococcusaureus, and Escherichia coli.
Mueller-Hinton Agar (Oxoi.
Bacto Agar (Difc.
Nutrient Agar (NA) media (Oxoi.
Nutrien Broth (NB) media (Oxoi.
, 80% Methanol, 1% H2SO4 solution, 1% BaCL2 solution.
Cloramphenicol antibiotic 10 g (AMP .
(Oxoi.
, and aquadest.
The equipment used in this study included: blender, oven, autoclave, centrifuge, vaporizer (Evaporator Butchi .
, shaker, vortex, digital scale, petri dish, laminar air flow, incubator.
Erlenmeyer flask, beaker glass, electric stove, sterile swab, tweezers, measuring flask, volumetric pipette, filter paper, bunsen burner, test tube, stirring rod, loop needle, aluminum foil, measuring cup, cotton swab, lighter, label paper, marker pen, paper disc, pH universal, bulb, and millimeter ruler.
Procedure Preparation of Growth Medium and Sterilization of Equipment NA (Nutrient Aga.
Nutrien Agar (NA) media was prepared by: weighing 23 grams of Nutrient Agar powder, adding 5 grams of Bacto Agar, then dissolving in 1 L of distilled water, stirring until the mixture was smooth while heated until boiling.
After that, it was put into a prepared test tube of as much as 5 mL for oblique culture, and the rest was put into an Erlenmeyer.
Then covered with cotton, sterilized in an autoclave at 121 oC, pressure 1-2 atm for 15 minutes.
NB (Nutrient Brot.
Nutrient Broth (NB) media was prepared by: weighing 23 grams of sodium broth powder, then dissolved in 1 L of distilled water, stirred until the mixture was smooth while heated until boiling.
After that, it was put into a prepared test tube of 5 mL as many as 10 tubes and the rest was put into an Erlenmeyer.
Then covered with cotton, sterilized in an autoclave at 121oC, pressure 1-2 atm for 15 minutes.
Mueller-Hinton Agar This medium was made by weighing 38 grams of Mueller-Hinton Agar (MHA), adding 6 grams of Bacto Agar, then dissolved in 1 L of distilled water, and heated to The media was then sterilized by autoclaving for 15 minutes at 121 oC .
-2 atm After sterilization, the media from the autoclave was allowed to stand until the temperature reached 45oC and poured into sterile Petri dishes.
This medium is used to test bacteria.
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Tool Sterilization Petri dishes, test tubes.
Erlenmeyer, and other glassware to be used are sterilized in an oven at 180oC for 2 hours after previously being washed, dried, and wrapped in Sample Preparation Medicinal plants Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
, first dried in an oven at 50oC, then crushed using a blender until smooth, then sieved using the smallest size sieve with a diameter of 150 mesh.
Extraction of medicinal plants Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
The powdered sample was extracted by: weighing 100 grams of plant powder and then adding 500 ml of 80% methanol as a solvent, then the sample was shaken for 12 hours and macerated for 24 hours at room temperature.
The macerated plant extract was filtered using filter paper to form two parts in the form of filtrate and dregs.
Furthermore, the dregs were added with 250 ml of methanol and shaken, then macerated again as in the previous work, this was done three times.
The extracts were mixed and centrifuged at 3500 rpm for 15 minutes and then filtered again.
The resulting supernatant was evaporated at a temperature of 40 oC with a vacuum rotary evaporator until it was dry which was marked by the presence of Furthermore, it is used to test the antibacterial activity.
Preparation of Test Bacteria A total of one ose colony of test bacteria (Staphylococcus aureus and Escherichia col.
was dissolved in 0.
85% physiological NaCl solution and the number of test bacteria used was homogenized using the McFarlan standard 0.
acterial 5 x .
on a black background, and bright light.
Mc.
Farlan turbidity standard 5 was prepared by mixing 0.
5 ml of 1% BaCL2 solution with 9.
5 ml of 1% H2SO4.
The sterile swab was dipped into a mixture of test bacteria with 0.
physiological NaCl, then drained by pressing the end of the swab, and rotating it against the inner wall of the tube to remove excess fluid.
Then the swab is applied to the surface twice, namely horizontally and vertically so that bacterial growth can be evenly Antibacterial Test The antibacterial activity test was carried out using the Kirby-bauer .
diffusion method.
Each blank paper disc was heated in an oven at 70 oC for 15 minutes.
Then each disc was given a solution of plant extract with a predetermined concentration .
5%) with a 0.
25 L micropipette.
The discs were allowed to stand for 15 minutes before being placed on the test medium.
Ampicillin 10AAg antibiotics and 80% methanol for each test bacteria were used as positive and negative controls.
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The Petri dishes were then incubated at 37 oC for 24 hours.
The diameter of the inhibition zone .
arked by the formation of a clear zon.
formed around the disc was measured using a ruler .
In measuring the inhibition zone, concentrations with a diameter of 6 mm .
qual to the diameter of the dis.
were declared to have no inhibition zone, while concentrations with a diameter of more than 6 mm were declared to have an inhibition zone.
Research Design The design used in this study was a factorial completely randomized design (CRD), as factors were the concentration of medicinal plant extracts .
chloramphenicol as a positive control and methanol as a negative contro.
and 4 types of bacteria.
test (E.
Salmonella typhi.
Bacillus cereus, and S.
The antibacterial test was carried out with 3 replications.
The number of treatments was 60 studies consisting of 4 types of test bacteria (E.
coli, and S.
x 3 concentrations of macrofungi extract .
5%) and 2 controls .
ositive control ampicillin and methanol.
negative contro.
The inhibition zone formed was observed as a RESULT Inhibition Zone Diameter of Reundeu Caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
plant extracts against four types of bacteria tested.
Escherichia coli and Salmonella typhi bacteria.
The ability of Reundeu Caret and Honje plant extracts to inhibit the growth of E.
coli and S.
Typhi bacteria is shown in Table 1.
Table 1.
Average Inhibition Zone Diameter of Plant Extracts and Honje against Eschericia coli and Salmonella typhi bacteria Test Bacteria Plant Extract Reundeu caret (Staurogyne Honje (Etlingera Extract Concentration (%) Average Inhibition Zone Diameter .
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Control Reundeu caret (Staurogyne Honje (Etlingera Control Control Information: Control = Kloramphenicoi Control - = Metanol 70% 6 mm = No growth inhibition zone Staphylococcus aureus and Bacillus cereus bacteria The ability of Reundeu Caret and Honje plant extracts in inhibiting the growth of S.
aureus and B.
cereus bacteria is shown in Table 2.
Table 2.
Average Inhibition Zone Diameter of Plant Extracts and Honje against S.
aureus and B.
cereus bacteria.
Test Bacteria Plant Extract Reundeu caret (Staurogyne longat.
Honje (Etlingera Extract Concentration (%) Control Control Reundeu caret (Staurogyne longat.
Honje (Etlingera Average Inhibition Zone Diameter .
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Control Control Description: Control = Chloramphenicoi Control - = Metanol 70% 6 mm = No growth inhibition zone Figure 1.
Antibacterial Bioactivity of Rendeu caret and Honje plants against the growth of E.
coli and S.
typhi bacteria Description: A.
Escherichia coli bacteria B.
Salmonella typhi bacteria Figure 2.
Antibacterial Bioactivity of Rendeu caret and Honje plants Journal of Tropical Biodiversity.
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against the growth of S.
aureus and B.
cereus bacteria Description: A.
aureus bacteria B.
cereus bacteria The inhibition zone of Rendeu Caret and Honje plants against B.
cereus bacteria was greater than that of S.
aureus, but the difference was not too significant (Figure .
Ekstrak Rendeu Caret Ekstrak Honje Figure 3.
Comparison of Inhibitory Zones of Rendeu Caret and Honje plant extracts cereus and S.
aureus bacteria Description: A.
Growth of S.
aureus bacteria B.
Growth of B.
cereus bacteria Effect of Concentration of Rendeu Caret Plant Extract and Honje Plant Extract on the Growth of Test Bacteria The results of the observation of the antibacterial activity test of Reudeu Caret and Honje plant extracts with different concentrations against the test bacteria S.
and Bacillus cereus showed that there was an effect of concentration treatment on the growth of these bacterial colonies (Figure .
, but there was no effect on the bacteria coli and S.
The average inhibition zone of the Rendeu Caret and Honje plant extracts with different concentrations against B.
cereus and S.
aureus bacteria can be seen in Figure 4.
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Reundeu caret Series1 Honje Series2 Series3 Series4 Figure 4.
Bar Diagram of Average Inhibitory Zone Diameter .
Rendeu Caret and Honje Plant Extracts with Different Concentrations Against Staphylococcus aureus and Bacillus subtilis Bacteria Description: Series 1= 50% Series 2 = 25% Series 3= 12,5% Series 4=6,25% DISCUSSION Table 1 and Table 2 above show that the extracts of the Rendeu caret plant and the Honje plant were only able to inhibit the growth of S.
aureus and B.
cereus bacteria.
The ability of these plant extracts to inhibit the growth of S.
aureus and B.
bacteria because the active compounds produced by these plants are able to damage the test bacterial cells, this can be seen from the inhibition zone produced which is larger than 6 mm (Figure .
, while for bacteria E.
coli and Salmonella typhi did not form an inhibition zone at all .
he diameter of the inhibition zone was only 6 m.
(Figure .
According to Jackie et al.
and Mariani .
, the main bioactive compounds contained in the two plants are.
saponins, flavonoids, and steroids/triterpenoids.
Saponins are bioactive compounds produced by certain plants.
These compounds are antibacterial, can interfere with the permeability of microbial cell membranes, resulting in cell membrane damage so that various important components from inside microbial cells, such as proteins, nucleic acids, nucleotides, and others can come out of the cell (Astuti et al.
, 2.
Furthermore.
Umar et al.
stated that microbial cells in this case bacteria can be damaged by bioactive compounds such as flavonoids produced by plants.
These flavonoids contain phenolic compounds which is acidic alcohol, known as carbolic acid.
Phenol works to denature proteins and damage cell membranes, phenol binds to proteins through hydrogen bonds which causes the protein structure to be damaged.
Bacteria S.
aureus and B.
cereus are two examples of bacteria that belong to the group of Gram-positive bacteria, while E.
coli and S.
typhi are a group of Gram29 Journal of Tropical Biodiversity.
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negative bacteria.
These two groups of bacteria have different cell wall structures.
The cell wall of Gram-positive bacteria consists of one layer, a thick layer of peptidoglycan, and a thin fat layer of 1-4%, while Gram-negative bacteria consist of 3 layers, namely lipoproteins, outer membrane phospholipids, and lipopolysaccharides, with lipid content ranging from 11%- 22%.
The constituent components of the layered Gram-negative cell walls, especially the phospholipid content, make it difficult for chemical components .
ioactive compound.
that are antibacterial to penetrate the cell walls of these Gramnegative bacteria (Poeloengan and Praptiwi, 2.
The inhibition zone of Rendeu Caret and Honje plants against B.
cereus bacteria was greater than that of S.
aureus, but the difference was not too significant (Figure .
According to Jawetz et al .
, the size of the diameter of the inhibition zone is what determines the potential of an antibacterial compound.
The data shown in Figure 4 shows an increase in the inhibition zone of the Rendeu Caret and Honje plant extracts against S.
aureus and B.
cereus bacteria in line with the increase in the concentration of the extract.
The smallest average inhibition zone .
5 m.
resulted from treatment using an extract concentration of 6.
25% against aureus and B.
cereus bacteria, and the largest average inhibition zone .
50 m.
was formed by treatment using an extract concentration of 50% against B.
The increase in the diameter of the inhibition zone for the growth of the test bacteria was due to the increasing number of plant extracts given.
According to Jawetz et al .
, the higher the concentration of an extract, the higher the inhibitory ability of an extract, then according to Kavitha et al.
and Ramadani .
, increasing the concentration of an antibacterial substance, will increase the content of active compounds produced by these substances so that the antibacterial activity is greater.
When observed from the average inhibition zone produced and associated with the grouping of antibacterial activity, the antibacterial activity of these two plant extracts was classified as moderate to strong, which was 6.
5 - 10.
According to Morales et al.
, antibacterial activity by active ingredients was grouped into 4 categories, namely weak activity (<5 m.
, moderate .
-10 m.
, strong .
-20 m.
, and very strong (>20-30 m.
Standard antibiotic Chloramphenicol 30AAg formed an inhibition zone for the growth of the four colonies of test bacteria.
The average inhibition zone formed by chloramphenicol 30AAg against E.
coli was 33 mm, against S.
typhi by 31 mm, against aureus by 38 mm, and against B.
cereus by 24 mm.
Based on these data, it is known that the antibacterial properties contained in the plant extracts of Rendeu Caret and Honje are not as effective as the standard antibiotics used.
Antibiotic Chloramphenicol 30 g affects four types of bacteria, both Gram-positive and Gram-negative, this antibiotic has a broad spectrum.
Meanwhile, the plant extracts of Rendeu Caret and Honje were only able to inhibit the growth of Gram-positive bacteria, so it can be said to have a narrow spectrum.
CONCLUSION
The results of the study entitled "Antibacterial Bioactivity of Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
Plant ExtractsAy, several conclusions can be drawn.
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Reundeu caret (Staurogyne longat.
and Honje (Etlingera hemisphaeric.
plant extracts were able to inhibit the growth of Staphylococcus aureus and Bacillus cereus bacteria, and were unable to inhibit the growth of Escherichia coli and Salmonella typhi bacteria.
There was an increase in the inhibition zone in line with the increase in the concentration of the extract.
Antibacterial activity The Honje (Etlingera hemisphaeric.
plant extract was not much different from the Reundeu caret (Staurogyne longat.
extract in inhibiting the growth of Staphylococcus aureus and Bacillus cereus bacteria.
The antibacterial activity of Rendeu Caret and Honje plant extracts was moderate to ACKNOWLEDGMENT Thank you to the Universitas Nasional and LPPM Unas who have assisted in funding this research, and also to the Microbiology and Genetics laboratory staff who have helped during research in the Laboratory.
REFERENCES