Online-ISSN 2565-1409 Journal of Widya Medika Junior Vol 3. No. 2 April 2021 ANTIBACTERIAL EFFECT OF GNETUM GNEMON L. LEAVES EXTRACT ON STAPHYLOCOCCUS AUREUS Paul J Dayoh1), Endang Isbandiati2), Titien Rahayu3) ABSTRACT Introduction: Bacterial infections are common among doctors in Indonesia, leading to increased use of antibiotics. Increased use of antibiotics, if not accompanied by rational indication, can increase bacterial resistance. At present, alternative medicine from natural ingredients is widely studied, one of which is the leaves of melinjo (Gnetum gnemon L.). Melinjo leaves are thought to contain antibacterial compounds such as flavonoids, tannins, saponins, and alkaloids. Staphylococcus aureus is a round Gram-positive bacteria, that forms grape-like clusters. Staphylococcus aureus is a normal flora on the skin and human mucosa but can be opportunistic pathogens that cause mild to severe infections such as sepsis. Pathogenic Staphylococcus aureus can produce hemolysis blood, frozen plasma, and produce various extracellular enzymes and toxins. Purpose: This research aims to study the MIC and MBC of melinjo leaves extract on the growth of Staphylococcus aureus bacteria. This research is an experimental study with a post- test control group design. Method: Testing conducted using the microdilution method. Melinjo leaves used were extracted by maceration method using 96% ethanol. Samples were Staphylococcus aureus ATCC 25923 from BBLK Surabaya. Results: extract of melinjo leaves (Gnetum gnemon L.) had MIC on Staphylococcus aureus bacteria at a concentration of 640 mg/ml. MBC was not obtained because the extract of melinjo leaves clotted above 1280 mg/ml concentration. Conclusion: Melinjo leaves extract has an antibacterial effect against Staphylococcus aureus, the MIC is obtained at a concentration of 640 mg/ml, and the MBC cannot be determined. Keywords: Antibacterial, Gnetum gnemon L., Staphylococcus aureus, Microdilution Minimum Inhibitory Concentration, Minimum Bactericidal Concentration 1)Student of Faculty of Medicine, Widya Mandala Surabaya Catholic University Indonesia, Jl. Kalisari Selatan No.1 Surabaya Email: [email protected] 2)Clinical Pharmacology Department, Faculty of Medicine, Widya Mandala Surabaya Catholic University Indonesia, Jl. Kalisari Selatan No.1 Surabaya 3)Clinical Pathology Department, Faculty of Medicine, Widya Mandala Surabaya Catholic University Indonesia, Jl. Kalisari Selatan No.1 Surabaya 122 Antibacterial Effect of… Dayoh PJ, Isbandiati E, Rahayu T INTRODUCTION resistance, including the selection of In Indonesia, bacterial infections are antibiotics based on local sensitivity common among clinicians, both in patterns, the use of appropriate and careful hospitals and private practice, which also catheters, proper antiseptic techniques, causes an increase in the use of antibiotics. infection control procedures to isolate Data from the Proceedings of the National pathogens, patient compliance with Academy of Sciences of the United States antibiotic use, and hand sanitation, and of America (PNAS 2018) states that in always pay attention to the cleanliness of 5 2000-2015 there was an increase in the use the environment around the patient . of antibiotics in the world by 65%, Infection is the process of especially in the United States, France, and microorganisms entering the body, Italy in the broad-spectrum penicillins, multiplying and causing severe health cephalosporins, quinolones, and problems. Bacteria can enter the body macrolides class1. World Health through wounds on the mucous Organization (WHO 2015) states that membranes, respiratory tract, digestive 6 about 64% of antibiotics are purchased tract, or genital and urinary tracts . One of without using a prescription in Southeast the bacteria that can infect humans is Asia. High rates on the use of antibiotics Staphylococcus aureus. Staphylococcus is without a prescription in Southeast Asia round-shaped Gram-positive bacteria with are caused by low public awareness a diameter of 0.7 to 1.2 µm, that do not regarding appropriate use of antibiotics2. move, do not produce spores, and are 7 Irrational use of antibiotics can arranged in grape-like clusters . Some increase bacterial resistance rates. Staphylococcus species are normal flora on According to the Centers for Disease the skin and human mucosa, while other Control (CDC 2013), there are around species can cause pus, abscess formation, 2,049,442 cases of illness due to antibiotic various pyogenic infections, and even fatal resistance and about 23,488 cases of septicemia. Pathogenic Staphylococcus can death3. Some theories of resistance include produce hemolysis blood, freeze plasma, the production of enzymes that can and produce multiple extracellular deactivate antibiotics, changes in the enzymes and toxins. The toxin produced structure of target receptors, and decreased by Staphylococcus is an enterotoxin that is bacterial cell membrane permeability for stable to heat and can cause food 6 antibiotics4. The CDC has made several poisoning . recommendations for reducing antibiotic 123 Online-ISSN 2565-1409 Journal of Widya Medika Junior Vol 3. No. 2 April 2021 Indonesia is a country with a lot and cytoplasm of Enterobacter cloacae variety of natural resources8. According to cells, thus interfering with the exchange of the Republic of Indonesia Health Office nutrients and metabolites and ultimately (2007), an estimated 9,600 species grow in inhibiting the supply of energy for Indonesia and have medicinal properties. bacteria12,13,14,15,16. Anti-bacterial activity Still, only around 300 species have been against Staphylococcus aureus shows that used as traditional medicine. One of them flavone, a flavonoid compound, is melinjo plant. Melinjo leaves and fruit significantly suppresses the production of are often processed into vegetables, and staphyloxanthin and α-hemolysin17. melinjo fruit seeds are processed into Tannins are phenolic compounds that can chips. parts of melinjo seeds are often used interfere with iron absorption by forming in herbal medicine research as natural anti- complex bonds with iron, thereby reducing bacterial. According to Siswoyo (2007), the bioavailability of iron in bacteria18. anti-bacterial from melinjo plants can be Saponin has an anti-bacterial mechanism found in the seeds, skins, and leaves of by disrupting the balance of surface melinjo plants9. permeability of bacterial cell membranes. phytochemical examination of melinjo Cell membrane damage can occur and leaves found that melinjo leaves contain causes various important components to several anti-bacterial compounds including come out of the bacterial cell, such as flavonoids, tannins, saponins, and proteins, nucleic acids, and nucleotides19. alkaloids10,11. The alkaloid anti-bacterial effect has Flavonoids are very lipophilic several mechanisms, first by inhibiting the compounds that make it possible to synthesis of nucleic acids because they damage bacterial membranes. The anti- inhibit the enzyme dihydrofolate reductase, bacterial effect of flavonoids is suspected secondly by interrupting the Z-ring, to involve three mechanisms: First, by thereby inhibiting cell division and inhibiting nucleic acid synthesis. Second, inhibiting the synthesis of nucleic acids. by damaging the cytoplasmic membrane, Finally, penetration of the outer membrane For example, the addition of galangin to and distruption of the integrity of the Staphylococcus aureus bacteria cause bacterial cytoplasmic membrane, which potassium leakage. Finally, by inhibiting causes intracytoplasmic leakage20. the metabolism of energy in bacteria. Research on the use of melinjo leaves Examples of administration of apigenin as an anti-bacterial is still minimal in data and naringenin alter the outer membrane and scientific sources. Therefore we are 124 Antibacterial Effect of… Dayoh PJ, Isbandiati E, Rahayu T interested in examining the anti-bacterial concentrated using Waterbath to get a thick effect of extracts of melinjo leaf (Gnetum extract. The thick extract was obtained gnemon L.) on Staphylococcus aureus. then the yield was calculated. METHOD The Making of Extract concentrations This research is an experimental study The results of ethanol extract of with a post-test control group design. This melinjo leaves obtained after maceration, research was conducted at the then diluted with Dimethyl sulfoxide 2% Phytochemical-Pharmacogenomic (DMSO) to form concentrations of 40 Laboratory of the Faculty of Pharmacy mg/ml, 80 mg/ml, 160 mg/ml, 320 mg/ml, Widya Mandala Catholic University and 640 mg/ml. Surabaya, the Surabaya Health Laboratory Center (BBLK) in Faculty of Pharmacy Bacteria Preparation Research Laboratory in Widya Mandala Staphylococcus aureus ATCC 25923 Catholic University Surabaya. The bacterial culture was regenerated to obtain population of this research is bacterial culture in the growth phase. Staphylococcus aureus. The sample used Regeneration was carried out on Mueller- was the bacterium Staphylococcus aureus Hinton broth media and then incubated for ATCC 25923. The concentration of 24 hours at 37ºC in an incubator. melinjo leaf extract used was 40-640 Microscopic and biochemical tests mg/ml. (catalase tests, coagulase tests, and mannitol tests) were carried out to ensure The Making of Extracts that the bacteria used are not contaminated. Melinjo leaf powder that has been obtained from UPT Batu City Medical Antibacterial Test Material was taken to the Phytochemical- Antibacterial activity testing was carried Pharmacogenomic