Piper Betel Leaf Research Articles

Application of piper betel leaf (piper betle linn) extract to control fish pathogenic bacteria in-vitro

Betel leaf extracts have some natural antimicrobial. This extracts has been used as antimicrobial agent in some aqua farmer in Indonesia for an alternative method to control fish diseases. This study aims to determine the effects of betel leaf extract on fish pathogenic bacteria in-vitro. Three kinds of solvents were used in betel leaf extracts. There were ethanol, n-hexane and ethyl-acetate. About 100 g of betel leaf simplicia were diluted by 1000 ml of solvents in stainless container cap for 24 hours maceration. This extracts were filtered with 0.42 μm paper filter and then evaporated at 50C during 30 minutes in 50 rpm with vacuum rotary evaporator. This concentrated extracts were used to antimicrobial testing. The pathogenic bacteria for in-vitro testing were from Gram-negative bacteria such as Aeromonas hydrophila, Vibrio alginolyticus and Edwardsiella tarda. The in-vitro test of herbal extract from betel leaf againts Gram-negative pathogenic bacteria indicated that the betel leaf extract could inhibited the growth of pathogenic bacteria with diameter mean of inhibition zone were more than 14 mm. The solvents variations were not significantly different (p > 0.05) to diameter inhibition zone of V. alginolyticus and A. hydrophila but significantly different (p < 0.05) to E. tarda.

Combination of Piper betel Leaves and Areca catechu Nuts Ethanolic Extract Effects on In Vitro Antimicrobial Test against Aerobic Mouth Microbiota

Introduction: Oral health awareness in Indonesia is still low with more and more oral and/or dental infection becomes more prevalent such as gingivitis, endodontitis, and pericoronitis. These infections could be caused by mouth microflora such as lactobacilli, Streptococcus mutans, Streptococcus mitis, Streptococcus oralis, and Streptococcus sobrinus. As time goes by, these bacteria could also become more problematic as antibacterial resistance emerges. Betel nut and betel leaf are well known traditional herbs that is often used in combination for "nyirih"; an Indonesian tradition to chew both plants often after meal. It is believed that these plants treat bad breath and also increases oral health as it could inhibit bacterial growth. Method: Betel nut and betel leaf were dried, powdered and macerated using 96% ethanol. The extract was tested on mouth microflora of volunteers grown in BHI using agar using disk diffusion test and modified E-test strip. Results: The betel leaves extract showed antibacterial activity in most of concentration used (1.25%, 2.5%, 5%, 10%) while betel nuts only gave antimicrobial effect in 10% concentration. The E-test results also showed the synergistic action between Piper betel leaves and Areca catechu nuts with some giving significant difference compared to the single treatment (P&lt;0.05). Conclusion: Piper betel leaves gives antimicrobial effect with MIC value of 1.25% while Areca catechu have MIC value of 10%. Furthermore, combination of both resulted in synergistic effect as indicated with larger diameter of inhibition compared to the single treatments.

Effect of Piper betel leaf extract on learning and memory in Aluminium chloride induced Alzheimer’s disease in Wistar rats

Alzheimer's disease is a common neurological disorder affecting a significant proportion of the elderly population.There are only a few drugs which can be used safely to treat it. We sought to investigate for the first time Piper betel leaf, a postmeal mouth freshener for its potential use in Alzheimer's disease. Five groups of male Wistar rats with six rats in each group aged 10-12 weeks were used. The control group received the distilled water, aluminium chloride (AlCl3) group was treated with AlCl3, the standard group was treated with rivastigmine with AlCl3, and the two test groups received piper betel leaf extract (PBE) at doses of 400mg/kg and 500mg/kg body weight with AlCl3. AlCl3 was administered orally for 42 days in all the treated groups. Memory and learning were evaluated by Morris water maze test and Passive avoidance test.The results of the Morris water maze test showed reduced mean escape latency period in all the groups on trial day three (P≤0.05) and on trial day four (P ≤0.01) compared to AlCl3 group. The retention of spatial memory by probe trial showed that PBE and rivastigmine treated group spent more time in the target (platform) quadrant when compared to AlCl3 group (P≤0.01). The passive avoidance test showed a significant increase in step through latency in standard and test groups compared to the AlCl3 treated group. The weight of the rats treated with AlCl3 and PBE was reduced at the end of the treatment period while increased in standard and control group. The study shows the beneficial effects of Piper betel leaves in Alzheimer’s disease by significantly improving the learning and memory functions in rats.

Antispermatogenic Effect of Piper betel Leaf Stalk Extract with Reference to Kinetic Studies of 17β-hydroxy Steroid Dehydrogenase Enzyme in Testes of Albino Rats

Background: In the development and maintenance of male reproductive function and fertility, steroidogenesis plays a key role. Aims: The aim of the present study was to investigate the effect of the betel leaf stalk extract on 17β- hydroxy steroid dehydrogenase activity levels. Methods: The observed elevation in testicular cholesterol levels may be due to decreased androgen production, which resulted in impaired spermatogenesis. The decreased steroidogenic enzyme 17β- HSD activity represents decreased androgen production by the extract administration. Reduction in enzyme active site density and Km value revealed that there was a reduction in enzyme-substrate affinities and rate of E-S complex breakdown in the administered rat testes. Results &amp; Conclusion: The administration of betel leaf stalk extract resulted in decreased enzyme content probably through impaired synthesis.

Hydroxychavicol sensitizes imatinib-resistant chronic myelogenous leukemia cells to TRAIL-induced apoptosis by ROS-mediated IAP downregulation.

The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a member of cytokine superfamily, induces apoptosis in a number of tumor cells through the activation of extrinsic apoptotic pathway but shows little or no cytotoxicity toward normal cells. However some tumor cells are inherently resistant to TRAIL-mediated apoptosis, which needs to be addressed to establish TRAIL as a potential chemotherapeutic drug. In this study, our aim was to manipulate TRAIL-apoptosis pathway by hydroxychavicol (HCH), a polyphenol from Piper betel leaf, for the induction of apoptosis in TRAIL resistant chronic myeloid leukemia cell. When imatinib-resistant K562 cells were treated with HCH, it made these K562 cells sensitive to TRAIL. It was observed that HCH downregulated antiapoptotic proteins XIAP and FLIP, whereas the expression of TRAIL receptors, DR4 and DR5, remains unchanged. Moreover, we observed that reactive oxygen species or ROS played a crucial role in the downregulation of FLIP and XIAP because ROS scavenger significantly reversed the decrease of XIAP, and FLIP. Ubiquitin-proteasome pathway was observed to play a crucial role in the downregulation of XIAP and FLIP, as proteasomal inhibitor MG132 significantly reversed the downregulation of XIAP and FLIP. In conclusion, this study demonstrates the combinatorial treatment of TRAIL and HCH as promising alternative therapeutic approach to treat the imatinib-resistant leukemia, which are also resistant to TRAIL.

The Effect of Rice Hull in Diets on Performance, Antioxidant Capacity and Blood Chemical Profile of Bali Duck

Abstract Background and Objective: An experiment was conducted to study the effects of diets with rice hull urea deammoniated and biofermentedbyacomplexoflactobacillusbacteriawithPiperbetleL.leaf supplementontheperformance,antioxidantcapacityand blood chemical profile of male Bali ducks in the growth phase. Methodology: The experiment was a completely randomized design (CRD) with five treatments; the treatments were: (A) control treatment, (B) diet containing 10% rice hull, (c) a diet containing 10% ammoniated and biofermented rice hull, (D) a diet containing 10% rice hull and Piper betle leaf and (E) a diet containing 10% ammoniated and biofermented rice hull and Piper betle leaf. Each treatment consisted of five Bali ducks and each treatment was replicated four times. Observed variables were consumption and digestibility of diets, final body weight, body weight gain and feed conversion ratio (FCR). Results: The results showed that diets containing 10% rice hull ammoniated and biofermented did not affect feed consumption (p>0.05), although feed digestibility and final body weight increased (p 0.05). Diets containing rice hull deammoniated and biofermented with piper betel leaf supplement decreased concentrations in Bali duck blood. Conclusion: In conclusion, diets offered with ricehullureadeammoniatedandbiofermentedsupplementedwithPiperbetleleaf improvedtheperformance,antioxidantcapacity and blood chemical profile of Bali ducks in the growth phase.

Chavibetol corrects thyrotoxicosis through alterations in thyroid peroxidase

Thyrotoxicosis is a clinical syndrome that commonly results from excess secretion and/or release of thyroid hormones in the circulation. It affects most of the body systems and if not treated properly may lead to serious health problems. In this investigation, we isolated a phenolic compound, chavibetol (CHV) from Piper betel leaf and evaluated its possible ameliorative effects in thyrotoxicosis of rats. Adult female rats were rendered thyrotoxic by the administration of L-thyroxine (L-T4) at 500μg/kg/day, i.p., for 12days, and then chavibetol (20.0mg/kg, p.o.) was administered for 2weeks. L-T4 administration elevated the concentration of serum thyroxine and triiodothyronine, activities of alanineaminotransferase and aspartate aminotransferase, and decreased the thyrotropin level as well as the expression of thyroid peroxidase (TPO). Further, it increased the activities of hepatic 5'mono-deiodinase-I, glucose-6--phosphatase, sodium-potasium-ATPase, and lipid peroxidation, and depleted the cellular antioxidants. However, chavibetol treatment to thyrotoxic rats normalized almost all these indices including TPO and also preserved the integrity of thyroid tissues suggesting its potential to correct thyrotoxicosis. Effects of CHV were more or less similar to a conventional antithyroid drug, propylthiouracil (PTU).

In vitro anti-cancer activity of Piper betel leaf extract on HA -29 and its anti-oxidant activity

In vitro anti-cancer activity of Piper betel leaf extract on HA -29 and its anti-oxidant activity

Ultra-Morphological Changes of Trichophyton Rubrum Treated with Hydroxychavicol

Trichophyton rubrum is a common pathogenic fungal species that is responsible for causing infection on human skin, hair and nail. The antifungal-resistant strains complicate the treatment regime. Hydroxychavicol (HC) is one of the main compounds from Piper betel leaf that have antifungal potential and its mechanism of action has not been studied yet. The objective of this preliminary study to determine the antifungal properties of HC against T. rubrum using transmission electron microscope (TEM) on gross and ultrastructure of T. rubrum hypha. T. rubrum was treated with HC and miconazole (MI) at concentrations of 1.25, 2.5, 5 and 10 mg/mL for 1, 3, 5 and 7 days continuously. Generally, fungi structures became more severely damaged at increasing treatment duration. Microscopically, the fungi’s cell wall treated with HC showed a rough surface, shrinkage and demolition similar to the MI treated group. The fungi organelles were also demolished and disorganized. This study revealed that HC has the ability to inhibit T. rubrum growth and has potential to be an antifungal agent for skin infections.

INHIBITORY EFFECT OF RED PIPER BETEL LEAF ETHANOL EXTRACT (PIPER CROCATUM RUIZ AND PAV.) AGAINST TRICHOMONAS VAGINALIS TROPHOZOITES IN VITRO

Objective: The objective of this study is to determine the minimum inhibitory concentration (MIC) of red betel leaf ethanol extract against Trichomonas vaginalis trophozoites.Materials and Methods: The samples of red piper betel leaf were extracted with 70% ethanol using a maceration method. The phytochemical screening of ethanol extract was performed using the standard procedures. Isolation of T. vaginalis trophozoites from vaginal secretions was conducted using swab method, then suspending it into a saline solution sterile and followed by observing trophozoites motility using a microscope. While the anti-leucorrhea activity test and determination of MIC extract was done using macrodillution method. The inhibitory effect of each extract concentration to trichomonas trophozoites’ morphology was observed under a microscope using staining techniques by adding Giemsa dyes.Results: The extract of red piper betel leaves contains several secondary metabolites such as alkaloids, flavonoids, polyphenols, quinones, and saponins. Of the six vaginal secretions, four positive trichomoniasis secretions were obtained. The results of anti-leucorrhea activity of red piper betel ethanol extract showed that the number of inactive trophozoites was proportional to the increasing of extract concentration. The inactive trophozoites were characterized by the absence of its motility under microscope observation. The MIC value of red piper betel extract of T. vaginalis trophozoites was 5% w/v.Conclusion: It can be concluded that the ethanol extract of red piper betel leaves could be contribute as a natural and safe anti-leucorrhea candidate for in vivo study on trichomoniasis in the future.

Photocatalysis effect of a novel green synthesis gadolinium doped titanium dioxide nanoparticles on their biological activities

Abstract We report the synthesis of gadolinium doped titanium dioxide (GdT) nanoparticles (NPs) via hydrothermal method using a novel bioreductant Piper betel leaf. The physiochemical properties of green synthesized GdT NPs were characterized using various techniques. The results reveal that GdT NPs have attached with the biomolecules which were supporting evidence for the reduction and capping agents. GdT NPs have crystalline in nature and their estimated grain size is of about 5.45 nm by the Scherer’s formula. GdT NPs consist of well-dispersed agglomerates of grains with a narrow size distribution of about 4 nm to 7 nm and are having spherical in nature. GdT NPs are thermally stable. In addition, GdT NPs were subjected to antimicrobial and antioxidant assays. Herein, we observed that the GdT NPs show higher antibacterial activities against Staphylococcus aureus as compared with Escherichia coli and the minimum inhibitory concentration (MIC) value was found to be 25 μg mL−1 under UV irradiation condition for both the cases. Furthermore, antioxidant activities of GdT NPs were evaluated in vitro, using the 1, 1- diphenyl-2-picrylhydrazyl (DPPH) radical cation decolorization test. Results suggest that GdT NPs give promising antioxidant activity as compared to standard ascorbic acid and Piper betel leaf. This novel and efficient strategy will show the route for avoiding the use of toxic solvents and a promising green route to restrict for drug discovery from natural products. Thus, the present findings may shine in the field of green pharmaceuticals industries.

Comparative Evaluation of Antifungal Activity of Piper Betel Leaf Oil, Origanum vulgare Essential Oil and Fluconazole Suspension on Candida albicans − An In Vitro Study

Introduction: Oropharyngeal candidiasis is an opportunistic mucosal infection caused by Candida albicans. It usually responds to topical treatments such as clotrimazole troches, topical fluconazole, chlorhexidine mouthwash and nystatin suspension. Piper betel leaf oil and Origanum vulgare essential oil have shown some topical antifungal activity. Aim: To determine and compare the antifungal efficacy of piper betel leaf oil, O. vulgare essential oil and fluconazole suspension against C. albicans. Materials and Methods: The zone of inhibition was measured by the cup–plate diffusion method using 100 μl volume of piper betel leaf oil, O. vulgare essential oil and fluconazole suspension, which were pipetted into the wells of the inoculated Sabouraud’s dextrose agar plates. The zone of inhibition was measured in millimetres using Vernier calliper. Minimum inhibitory concentration (MIC) was determined by the broth macro-dilution test by pouring 1 ml of the respective concentrations of the test material to the individual test tubes along with 10 μl of the diluted test organism inoculum. Finally, MIC was calculated using a reflective viewer. Results: The zone of inhibition for O. vulgare essential oil (>40 mm) was more than fluconazole suspension (>35 mm). MIC of O. vulgare essential oil, piper betel leaf oil and fluconazole suspension was 1.6%, 0.4% and 0.8%, respectively. Conclusion: O. vulgare essential oil was found to be a more effective antifungal agent than piper betel leaf oil and fluconazole suspension.

Determination of ultra-morphological changes of Trichophyton rubrum treated with hydroxychavicol

Introduction: Trichophyton rubrum is a common pathogenic fungal species that is responsible for causing infection of human skin, hair and nail. The antifungal-resistant strains however, complicate the treatment regime. Hydroxychavicol (HC) is one of the main compounds from the Piper betel leaf that have antifungal potential and its mechanism of action has yet to be studied. This study was carried out to determine the antifungal properties of HC against Trichophyton rubrum using transmission electron microscope (TEM) on gross and ultrastructure of Trichophyton rubrum hypha. Methods: Broth dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of HC and miconazole (MI) against the Trichophyton rubrum (ATCC 28188). Trichophyton rubrum was treated with HC and MI at concentrations of 1.25, 2.5, 5 and 10 mg/mL for 1, 3, 5 and 7 days continuously. Results: The MIC results of the HC and MI against Trichophyton rubrum were 0.00048 mg/mL and 0.000061 mg/mL respectively. MFC results showed 0.0019 mg/mL for HC and 0.000061 mg/mL for MI. Microscopically, the fungi structures became more severely damaged at increasing treatment duration. The cell wall of the fungi treated with HC showed a rough surface, shrinkage and demolition similar to that of the MI treated group. The fungi organelles were also demolished and disorganized. Conclusions: This study reveals that HC has the ability to inhibit Trichophyton rubrum growth and it has the potential to be an antifungal agent especially in treating dermatitis.

Effect of Piper betel leaf stalk extract on protein metabolism in reproductive tissues of male albino rats

Article history: Received 18 July 2014 Objective: To know the impact of Piper betel leaf stalk (P. betel) extract on Protein and energy metabolism and its role in male albino rats. Methods: Healthy adult (3-4 months old) male Wistar strain albino rats were administered with betel leaf stalk extract, at the dose of 50 mg/ kg/day through oral gavages for 15 days. Twenty four hours after the last dose, the animals were autopsied. In order to assess antifertility effect in testis, epididymis, seminal vesicle and prostate gland, estimation of total, soluble and structural proteins, free amino acids and DNA, RNA were undertaken. Results: The accumulation in proteins indicates the anti-androgenic effect of extract. The reduction in free amino acids will affect the sertoli cell function, results in the damage of spermatogenesis. The significant elevation in testicular DNA content (hyperplasia) was observed. In the present study, P. betel leaf stalk extract decreases the concentration of RNA, in testes, seminal vesicle and prostate gland except in epididymis where it was elevated. It indicates the alterations in rate of protein synthesis and growth rate of tissues due to the administration of P. betel leaf stalk extraction. However, the RNA: DNA ratio was reduced except in prostate. Conclusions: P. betel leaf stalk extract exert its anti androgenic effect by alterations in rate of protein synthesis and cellular hypertrophy occur in prostate.

Inhibitory effect of Piper betel leaf extracts on copper-mediated LDL oxidation and oxLDL-induced lipid accumulation via inducing reverse cholesterol transport in macrophages

Piper betel leaf (PBL) has the biological capabilities of detoxification and can work as an anti-inflammatory agent and an anti-oxidant. In this study, we evaluated the anti-oxidative activity of the extract of Piper betel leaves (PBLs) on the basis of Cu2+-mediated oxidation, and its ability to prevent foam cell formation in a model for oxidised low density lipoprotein (oxLDL)-induced lipid accumulation in macrophages. Our data demonstrated that PBLs were able to inhibit LDL oxidation in vitro and are able to reduce the lipid accumulation in macrophages. We showed the underlying mechanisms to be the following: PBLs up-regulated the protein levels of the class A and class B scavenger receptors, the membrane lipid transporter ABCA1, and its upstream regulator Liver X receptor (LXR) in the macrophages exposed to oxLDL. The results suggested that PBLs activated the reverse cholesterol transport mechanism to enhance the metabolism of the oxLDL that could prevent both lipid accumulation and foam cell formation and further minimise the possible damage of vessels caused by the oxLDL.

Piper betel leaf extract: anticancer benefits and bio-guided fractionation to identify active principles for prostate cancer management

Plant extracts, a concoction of bioactive non-nutrient phytochemicals, have long served as the most significant source of new leads for anticancer drug development. Explored for their unique medicinal properties, the leaves of Piper betel, an evergreen perennial vine, are a reservoir of phenolics with antimutagenic, antitumor and antioxidant activities. Here, we show that oral feeding of betel leaf extract (BLE) significantly inhibited the growth of human prostate xenografts implanted in nude mice compared with vehicle-fed controls. To gain insights into the 'active principles', we performed a bioactivity-guided fractionation of methanolic BLE employing solvents of different polarity strengths using classical column chromatography. This approach yielded 15 fractions, which were then pooled to 10 using similar retention factors on thin-layer chromatographs. Bioactivity assays demonstrated that one fraction in particular, F2, displayed a 3-fold better in vitro efficacy to inhibit proliferation of prostate cancer cells than the parent BLE. The presence of phenols, hydroxychavicol (HC) and chavibetol (CHV), was confirmed in F2 by nuclear magnetic resonance, high-performance liquid chromatography and mass spectroscopy. Further, the HC containing F2 subfraction was found to be ~8-fold more potent than the F2 subfraction that contained CHV, in human prostate cancer PC-3 cells as evaluated by the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. Removing CHV from F2 remarkably decreased the IC50 of this fraction, indicating that HC is perhaps the major bioactive constituent, which is present to an extent of 26.59% in BLE. These data provide evidence that HC is a potential candidate for prostate cancer management and warrants further preclinical evaluation.

The antimicrobial effect of piper betel leaves extract against Streptococcus mutans

Introduction: Betel leaves are herbal plant that is believed to have phenolic compound as an antimicrobial agent. One of the medical plants (herbal) which has an-timicrobial properties is Piper betel.The aim of this research is to analyzed the antimicrobial effect of Piper Betel Leaves extract against Streptococcus mutans. Methods: Type of this research is laboratory experiment. Population of this study are Piper betle Linn plant in West Java. The MIC test were conducted based on serial dilution method in 64%, 32%, 16%, 8%, 4%, 2%, 1%, 0.5%, 0.25%, 0.125% concentration of Piper betel leaves extracts against 5 sample of Streptococcus mutans within three repetitions. Results: The result shows that 0.25% concentration of Piper betel leaves extract can inhibit Streptococcus mutans. Conclusion: Piper betel leaves extract can inhibit the growth and had antimicrobial effect against Streptococcus mutans.

Effects of ionic surfactants on the morphology of silver nanoparticles using Paan (Piper betel) leaf petiole extract

Stable silver nanoparticles were synthesized by the reduction of silver ions with a Paan (Piper betel) leaf petiole extract in absence and presence of cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulphate (SDS). The reaction process was simple and convenient to handle, and was monitored using ultraviolet-visible spectroscopy. Absorbance of Ag-nanoparticles increases with the concentrations of Paan leaf extract, acts as reducing, stabilizing and capping agents. The polyphenolic groups of petiole extract are responsible to the rapid reduction of Ag+ ions into metallic Ag0. The results indicated that the shape of the spectra, number of peaks and its position strongly depend on the concentration of CTAB, which played a shape-controlling role during the formation of silver nanoparticles in the solutions, whereas SDS has no significant effect. The morphology (spherical, truncated triangular polyhedral plate and some irregular nanoparticles) and crystalline phase of the particles were determined from transmission electron microscopy (TEM) and selected area electron diffraction (SAED).

IN-VITRO ANTIBACTERIAL ACTIVITY OF PIPER BETEL LEAF EXTRACTS

Ethanol, petroleum ether and chloroform extracts of Piper betel leaf were tested against Grampositive (Bacillus subtilis ATCC 6633), Staphylococcus aureus ATCC 29213, Micrococcus luteus ATCC 9341) and Gram-negative (Escherichia coli ATCC 25922, Pseudomonas putida, Salmonella typhi ATCC 19430, Shigella flexneri ATCC 25929, Pseudomonas aeruginosa ATCC 33347, Vibrio cholerae, Klebsiella pneumoniae, Proteus mirabilis ATCC 49565) bacterial strains by Agar-well Diffusion Method. All the crude extracts showed a broad spectrum of antibacterial activity inhibiting both the gram-positive and gram-negative bacteria. Petroleum ether extracts of Piper betel were found to be least effective against most of the tested organisms. Moderate antibacterial activity has been studied with chloroform extracts while ethanolic fractions have been investigated to show optimum activity against nearly all chosen strains. Levofloxacin, the semi-synthetic broad spectrum antibiotic was used as standard Compare to levofloxacin, bio active fractions of Piper betel showed maximum activity against the Klebsiella pneumonia. This study reports the possible activity of Piper betel leaf in arresting the growth of selected bacterial strains. It has been expected that the present work on antimicrobial isolation of the plant bio active parts will lead to the researchers who continue work that may bring clinical success concerning the fatal diseases.

Anti–microbial principles of selected remedial plants from Southern India

Objective To examine the anti–bacterial activity of leaf extracts of Morus alba L. ( Moraceae) and Piper betel L. ( Piperaceae), and seed extracts of Bombax ceiba L. ( Borabacaceae). Methods We have partially purified plant extracts by solvent extraction method, and evaluated the effect of individual fractions on bacterial growth using Escherichia coli ( E. coli), Pseudomonas aeruginosa ( P. aeruginosa) and Staphylococcus aureus ( S. aureus) bacterial strains. Results Compared with Morus and Bombax fractions, Piper fractions showed significant growth inhibition on all the three types of bacteria studied. The EtOAc-hexane fractions of Piper leaves exhibited significant anti–bacterial activity with minimum inhibitory concentrations (MIC) of 50 μg/ml culture against both gram-positive and gram-negative bacteria. The EtOAc-fractions I, II, and IV inhibited bacterial colony formation on soft agar in addition to growth inhibition. A combination treatment of piper fractions with ampicillin resulted in significant growth inhibition in E. coli and P. aeruginosa, and combination with anticancer drug geldanamycin (2μg/ml) showed selective growth inhibition against P. aeruginosa and S. aureus. Three major compounds, i.e., eugenol, 3-hexene-ol and stigmasterol, were primarily identified from Piper betel leaf extractions. Among the individual compounds, eugenol treatment showed improved growth inhibition compared with stigmasterol and 3-hexene-ol. Conclusions We are reporting potential anti–bacterial compounds from Piper betel against both gram-positive and gram-negative bacteria either alone or in combination drug treatment.