In vitro antibacterial and antibiofilm activities of Piper betle L. ethanolic leaf extract on staphylococcus aureus ATCC 29213

Background and Objective@#Staphylococcus aureus is the leading cause of skin and soft tissue infections such as abscesses, furuncles, and cellulitis. Biofilm forming strains of S. aureus have higher incidence of antimicrobial resistance to at least three or more antibiotics and are considered as multidrug resistant. Since S. aureus biofilm-producing strains have higher rates of multidrug and methicillin resistance compared to non-biofilm-producing strains, the need for alternative therapeutic option is important. Furthermore, rates of methicillin-resistant Staphylococcus aureus (MRSA) in Asia remain high. Results of the study may provide support for the clinical uses of P. betle as a topical antibacterial and antiseptic in the treatment and prevention of infections involving the skin, mouth, throat, and indwelling medical devices. Thus, this study aimed to evaluate the in vitro antibacterial and antibiofilm activities of Piper betle L. ethanolic leaf extract (PBE) against a biofilm-forming methicillin-sensitive Staphylococcus aureus ATCC 29213 (MSSA).@*Methods@#The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PBE against MSSA were determined using the agar dilution assay. The biofilm inhibition and eradication assays using crystal violet were done to quantify the antibiofilm activities of PBE on MSSA biofilm.@*Results@#PBE showed activity against MSSA in agar dilution assay with MIC and MBC values of 2500 μg/mL and 5000 μg/mL, respectively. At subinhibitory concentrations, PBE showed biofilm inhibition activity at 1250 μg/mL but a lower percent eradication of biofilms as compared to oxacillin was noted.@*Conclusion@#PBE showed antibacterial activities including biofilm inhibition against methicillin-sensitive Staphylococcus aureus ATCC 29213 (MSSA).

Antibacterial activity of Sireh (Piper betle L.) leaf extracts for controlling bacterial leaf blight diseases in rice plant

Aims@#This study aimed to determine the antibacterial activity of Piper betle L. leaf extract against Xanthomonas oryzae pv. oryzae that causes bacterial leaf blight in rice plant.@*Methodology and results@#The antibacterial activity of the P. betle leaf extract (100, 50, 25 and 12.25 mg/mL) with four different solvents (methanol, ethyl acetate, hexane and acetone) was evaluated using a disc diffusion assay, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. The inhibition zone of methanolic extract appeared to have the maximum diameter compared to those of other extracts, which is 32.67 mm at a concentration of 100 mg/mL, followed by 30.33 mm, 22.00 mm and 20.30 mm for the concentrations of 50 mg/mL, 25 mg/mL and 12.5 mg/mL, respectively. The MIC and MBC values of the methanolic extract were 0.625 mg/mL suggesting that the extract has a bactericidal effect on X. oryzae pv. oryzae (Xoo). The time-kill curve studies revealed that the 1× MIC (0.625 mg/mL) concentration of methanolic extract had a time and concentration-dependent killing effect on Xoo. Gas chromatography-mass spectrometry (GC-MS) analysis of methanol extract revealed the presence of eugenol acetate (29.53%), 4-allyl-1,2-diacetoxybenzene (29.51%) and 2,3-dimethyl benzoic acid (22.82%) as major compounds.@*Conclusion, significance and impact of study@#The methanolic leaf extract of P. betle was proven to have an effective inhibitory effect on Xoo and may have the potential to be used as an alternative management strategy for controlling rice diseases. In the future study, the methanolic leaf extract of P. betle is one of the recommendations to be applied in glasshouse and field trials.

Anti-migratory effects of Piper betle leaf aqueous extract on cancer cells and its microtubule targeting properties / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Piper betle (PB), also known as "betel" in Malay language, is a tropical Asian vine. PB leaves are commonly chewed by Asians along with betel quid. It contains phenols such as eugenol and hydroxychavicol along with chlorophyll, β-carotene, and vitamin C (Salehi et al., 2019). Extracts from PB leaves have various medicinal properties including anticancer, antioxidant, anti-inflammatory, and antibacterial effects (Salehi et al., 2019). Previous research has shown that PB induces cell cycle arrest at late S or G2/M phase and causes apoptosis at higher doses (Wu et al., 2014; Guha Majumdar and Subramanian, 2019). A combination of PB leaf extract has also been shown to enhance the cytotoxicity of the anticancer drug, 5-fluorouracil (5-FU), in cancer cells (Ng et al., 2014).

In Vitro Antibacterial and Antibiofilm activities of Piper betle L. Ethanolic Leaf Extract on Staphylococcus aureus ATCC 29213

Background and Objective@#Staphylococcus aureus is the leading cause of skin and soft tissue infections such as abscesses, furuncles, and cellulitis. Biofilm forming strains of S. aureus have higher incidence of antimicrobial resistance to at least three or more antibiotics and are considered as multidrug resistant. Since S. aureus biofilm-producing strains have higher rates of multidrug and methicillin resistance compared to non-biofilm-producing strains, the need for alternative therapeutic option is important. Furthermore, rates of methicillin-resistant Staphylococcus aureus (MRSA) in Asia remain high. Results of the study may provide support for the clinical uses of P. betle as a topical antibacterial and antiseptic in the treatment and prevention of infections involving the skin, mouth, throat, and indwelling medical devices. Thus, this study aimed to evaluate the in vitro antibacterial and antibiofilm activities of Piper betle L. ethanolic leaf extract (PBE) against a biofilm-forming methicillin-sensitive Staphylococcus aureus ATCC 29213 (MSSA).@*Methods@#The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PBE against MSSA were determined using the agar dilution assay. The biofilm inhibition and eradication assays using crystal violet were done to quantify the antibiofilm activities of PBE on MSSA biofilm. @*Results@#PBE showed activity against MSSA in agar dilution assay with MIC and MBC values of 2500 μg/mL and 5000 μg/mL, respectively. At subinhibitory concentrations, PBE showed biofilm inhibition activity at 1250 μg/mL but a lower percent eradication of biofilms as compared to oxacillin was noted. @*Conclusion@#PBE showed antibacterial activities including biofilm inhibition against methicillin-sensitive Staphylococcus aureus ATCC 29213 (MSSA).

Growth inhibitory response and ultrastructural modification of oral-associated candidal reference strains (ATCC) by Piper betle L. extract / 国际口腔科学杂志·英文版

Candida species have been associated with the emergence of strains resistant to selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease mucosal fungal infections. This study aimed to investigate the effects of Piper betle extract on the growth profile and the ultrastructure of commonly isolated oral candidal cells. The major component of P. betle was identified using liquid chromatography-mass spectrophotometry (LC-MS/MS). Seven ATCC control strains of Candida species were cultured in yeast peptone dextrose broth under four different growth environments: (i) in the absence of P. betle extract; and in the presence of P. betle extract at respective concentrations of (ii) 1 mg⋅mL(-1); (iii) 3 mg⋅mL(-1); and (iv) 6 mg⋅mL(-1). The growth inhibitory responses of the candidal cells were determined based on changes in the specific growth rates (µ). Scanning electron microscopy (SEM) was used to observe any ultrastructural alterations in the candida colonies. LC-MS/MS was performed to validate the presence of bioactive compounds in the extract. Following treatment, it was observed that the µ-values of the treated cells were significantly different than those of the untreated cells (P<0.05), indicating the fungistatic properties of the P. betle extract. The candidal population was also reduced from an average of 13.44×10(6) to 1.78×10(6) viable cell counts (CFU)⋅mL(-1). SEM examination exhibited physical damage and considerable morphological alterations of the treated cells. The compound profile from LC-MS/MS indicated the presence of hydroxybenzoic acid, chavibetol and hydroxychavicol in P. betle extract. The effects of P. betle on candida cells could potentiate its antifungal activity.

Inhibitory effect of Piper betle Linn. leaf extract on protein glycation - Quantification and characterization of the antiglycation components.

Piper betle Linn. is a Pan-Asiatic plant having several beneficial properties. Protein glycation and advanced glycation end products (AGEs) formation are associated with different pathophysiological conditions, including diabetes mellitus. Our study aims to find the effect of methanolic extract of P. betle leaves on in vitro protein glycation in bovine serum albumin (BSA)-glucose model. The extract inhibits glucose-induced glycation, thiol group modification and carbonyl formation in BSA in dose-dependent manner. It inhibits different stages of protein glycation, as demonstrated by using glycation models: hemoglobin-d-gluconolactone (for early stage, Amadori product formation), BSA-methylglyoxal (for middle stage, formation of oxidative cleavage products) and BSA-glucose (for last stage, formation of AGEs) systems. Several phenolic compounds are isolated from the extract. Considering their relative amounts present in the extract, rutin appears to be the most active antiglycating agent. The extract of P. betle leaf may thus have beneficial effect in preventing protein glycation and associated complications in pathological conditions.

Antioxidant enzyme activity and malondialdehyde levels can be modulated by Piper betle, tocotrienol rich fraction and Chlorella vulgaris in aging C57BL/6 mice

OBJECTIVE: The aim of this study was to determine the erythrocyte antioxidant enzyme activity and the superoxide dismutase, catalase, glutathione peroxidase, and plasma malondialdehyde levels in aging mice and to evaluate how these measures are modulated by potential antioxidants, including the tocotrienol-rich fraction, Piper betle, and Chlorella vulgaris. METHOD: One hundred and twenty male C57BL/6 inbred mice were divided into three age groups: young (6 months old), middle-aged (12 months old), and old (18 months old). Each age group consisted of two control groups (distilled water and olive oil) and three treatment groups: Piper betle (50 mg/kg body weight), tocotrienol-rich fraction (30 mg/kg), and Chlorella vulgaris (50 mg/kg). The duration of treatment for all three age groups was two months. Blood was withdrawn from the orbital sinus to determine the antioxidant enzyme activity and the malondialdehyde level. RESULTS: Piper betle increased the activities of catalase, glutathione peroxidase, and superoxide dismutase in the young, middle, and old age groups, respectively, when compared to control. The tocotrienol-rich fraction decreased the superoxide dismutase activity in the middle and the old age groups but had no effect on catalase or glutathione peroxidase activity for all age groups. Chlorella vulgaris had no effect on superoxide dismutase activity for all age groups but increased glutathione peroxidase and decreased catalase activity in the middle and the young age groups, respectively. Chlorella vulgaris reduced lipid peroxidation (malondialdehyde levels) in all age groups, but no significant changes were observed with the tocotrienol-rich fraction and the Piper betle treatments. CONCLUSION: We found equivocal age-related changes in erythrocyte antioxidant enzyme activity when mice were treated with Piper betle, the tocotrienol-rich fraction, and Chlorella vulgaris. However, Piper betle treatment showed increased antioxidant enzymes activity during aging.

Scanning Electron Microscopic study of Piper betle L. leaves extract effect against Streptococcus mutans ATCC 25175

INTRODUCTION: Previous studies have shown that Piper betle L. leaves extract inhibits the adherence of Streptococcus mutans to glass surface, suggesting its potential role in controlling dental plaque development. OBJECTIVES: In this study, the effect of the Piper betle L. extract towards S. mutans (with/without sucrose) using scanning electron microscopy (SEM) and on partially purifed cell-associated glucosyltransferase activity were determined. MATERIAL AND METHODS: S. mutans were allowed to adhere to glass beads suspended in 6 different Brain Heart Infusion broths [without sucrose; with sucrose; without sucrose containing the extract (2 mg mL-1 and 4 mg mL-1); with sucrose containing the extract (2 mg mL-1 and 4 mg mL-1)]. Positive control was 0.12 percent chlorhexidine. The glass beads were later processed for SEM viewing. Cell surface area and appearance and, cell population of S. mutans adhering to the glass beads were determined upon viewing using the SEM. The glucosyltransferase activity (with/without extract) was also determined. One- and two-way ANOVA were used accordingly. RESULTS: It was found that sucrose increased adherence and cell surface area of S. mutans (p<0.001). S. mutans adhering to 100 µm² glass surfaces (with/without sucrose) exhibited reduced cell surface area, fuffy extracellular appearance and cell population in the presence of the Piper betle L. leaves extract. It was also found that the extract inhibited glucosyltransferase activity and its inhibition at 2.5 mg mL-1 corresponded to that of 0.12 percent chlorhexidine. At 4 mg mL-1 of the extract, the glucosyltransferase activity was undetectable and despite that, bacterial cells still demonstrated adherence capacity. CONCLUSION: The SEM analysis confrmed the inhibitory effects of the Piper betle L. leaves extract towards cell adherence, cell growth and extracellular polysaccharide formation of S. mutans visually. In bacterial cell adherence, other factors besides glucosyltransferase are involved.

Comparative study of Candida in oral submucous fibrosis and healthy individuals / Estudo comparativo de Candida em fibrose submucosa oral e indivíduos saudáveis

PURPOSE: Oral submucous fibrosis (OSMF) is a high-risk precancerous condition that predominantly affects Indian youngsters due to the habit of gutkha chewing. Candida may play a role in the etiopathogenesis of premalignant and malignant lesions. The aim of this study is to compare the incidence, intensity, and species of Candida found in OSMF patients and healthy individuals. METHODS: This study included 20 OSMF patients and 20 healthy controls. A detailed history of each patient was recorded along with a clinical examination. Samples were collected with the oral rinse technique and cultured on Sabouraud's agar medium. The isolated yeast species were counted and identified based on Gram staining, a germ tube test, chlamydospore formation and a sugar assimilation test. RESULTA: In total, 40 percent of OSMF patients and 15 percent of healthy controls yielded Candida organisms on culture. C. albicans was the predominant species isolated, but C. krusei and C. tropicalis were also identified. Gender, gutkha habit and clinical staging had no influence on the candidal carriage in OSMF patients. CONCLUSION: The incidence and intensity of Candida (primarily C. albicans) was greater in OSMF patients than in healthy controls, but these findings were within the normal limit (3-47 percent). Therefore, Candida may not be an etiologic factor in malignant transformation. However, controversy still exists over whether the chewing of betel quid in cases of OSMF has an inhibitory effect or promotes the adherence and invasion of Candida.

OBJETIVO: Fibrose submucosa oral (FSO) é uma condição pré-maligna de alto risco que predominantemente afeta jovens da Índia devido ao hábito de mascar 'gutkha'. Candida pode ter um papel importante na etiopatogenia de lesões pré-malignas e malignas. O objetivo deste estudo foi comparar a incidência, intensidade e Candida spp encontradas em pacientes com FSO e em indivíduos saudáveis. METODOLOGIA: Este estudo incluiu 20 pacientes com FSO e 20 controles saudáveis. Obteve-se de cada sujeito uma história detalhada e exame clínico. As amostras foram coletadas com uma técnica de enxágue bucal e cultivadas em meio Agar Sabouraud. As espécies isoladas foram contadas e identificadas com base em coloração Gram, teste de tubo de ensaio e teste de assimilação de açúcar. RESULTADOS: No total, 40 por cento dos pacientes com FSO e 15 por cento dos controles saudáveis apresentaram resultado positivo de cultura para Candida. C. albicans foi a espécie predominante isolada, mas C. krusei e C. tropicalis também foram identificados. O sexo, hábito de mascar 'gutkha' e estadiamento clínico não influenciaram a presença de fungos nos pacientes com FSO. CONCLUSÃO: A incidência e intensidade de Candida (primariamente C. albicans) foi maior nos pacientes com FSO que nos sujeitos controle, mas estes achados estavam dentro dos limites normais (3-47 por cento). Portanto, Candida pode não ser um fator etiológico na transformação maligna. Entretanto, ainda há controvérsias se o hábito de mascar 'betel' em casos de FSO teria um efeito inibidor ou promoveria a aderência e invasão de Candida.

Alkaloids and lignans from stems of Piper betle / 中国中药杂志

Alkaloids and lignans from the stems of Piper betle were studied. Compounds were isolated and purified by repeated silica gel, reverse phase silica gel, Sephadex LH-20 column chromatography and preparative thin layer chromatography. The structures were elucidated on the basis of spectral analysis. From the ethyl acetate soluble fractions of the 70% acetone extract, ten compounds were isolated and identified as piperine (1), pellitorine (2), N-isobutyl-2E,4E-dodecadienamide (3), dehydropipernonaline (4), piperdardine (5), piperolein-B (6), guineensine (7), (2E,4E)-N-isobutyl-7-(3',4'-methylenedioxyphenyl)-2,4-heptadienamide (8), syringaresinol-O-beta-D-glucopyranoside (9),pinoresinol (10). All Compounds were isolated from the plant for the first time, and compounds 9 and 10 were isolated firstly from the genus.

Antioxidant effect of ethanolic extract of Piper betle Linn. (Paan) on erythrocytes from patients with HbE-beta thalassemia.

HbE-beta thalassemia is caused by an interaction between HbE and defective b globin gene of thalassemia. Repeated blood transfusions cause an iron overload, triggering an enhanced generation of free radicals. In the present study, the anti-oxidant property of ethanolic extract of the leaves of Piper betle Linn. (PB) was evaluated in the erythrocytes from patients with HbE-beta thalassemia. In patients with HbE-beta thalassemia (n = 30) and age- and sex-matched healthy individuals (n = 30), the baseline level of reactive oxygen species (ROS) and free radical scavenging activity in the erythrocytes was measured by flow cytometry using dihydrodichlorofluorescein diacetate (H2DCFDA), in terms of the geometric mean fluorescence channel (GMFC). The baseline generation of ROS was significantly higher in the erythrocytes from patients with HbE-beta thalassemia, as compared to healthy volunteers, the GMFC being 67.20 ± 4.64 vs. 23.03 ± 1.88 (p<0.001), which was effectively decreased by PB. Similarly, H2O2 (0.5-1.0 mM) induced a higher increase in the GMFC in the erythrocytes from patients with HbE-beta thalassemia, as compared to controls which was effectively reduced by PB. Taken together, PB showed promising anti-oxidant activity against the erythrocytes from patients with HbE-beta thalassemia.

effect of psidium guajava and piper betle extracts on the morphology of dental plaque bacteria

It has been reported that the aqueous extracts of Psidium guajava and Piper betle leaves showed anti-plaque activities during the early stages of dental plaque formation. The aim of the study was to elucidate if such anti-plaque activities involve any ultra-structural changes to the morphology of three early dental plaque bacteria, Streptococcus sanguinis, Streptococcus mitis and Actinomyces sp. Pure cultures of the bacteria were suspended in BHI medium and treated with the test herbal extracts at the sub-lethal concentrations. The growth mixtures were incubated at 37°C. At the logarithmic growth phase [t1], aliquots of 1 ml of the growth mixtures were fixed and used in the preparation of specimens for SEM studies. Ultra-structural alterations to the morphology of the treated cells noted were compared to those of the cells cultured under untreated conditions. Following exposure of the bacteria to the two test herbal extracts, profound ultra-structural changes to their morphology were observed. The observed structural or morphological alterations could attribute to the bacteria being less active in performing normal physiological metabolic functions and thus rendering them less efficient to multiply. The changes noted included [i] reduced sizes of the bacteria, and [ii] majority cells at the non-dividing state as compared to those cultured under controlled conditions. This study has shown anti-plaque effects of aqueous extract of both P. betle and P. guajava

Chewing of betel, areca and tobacco: perceptions and knowledge regarding their role in head and neck cancers in an urban squatter settlement in Pakistan.

The link of betel, areca and chewable tobacco with head and neck cancers is clearly established. Fifty eight percent of the global head and neck cancers occur in South and Southeast Asia, where chewing of betel, areca and tobacco are common. This study was carried out to establish the pattern of use of Paan, Chaalia, Gutka, Niswar, Tumbaku and Naas among population of squatter settlement of Karachi and to determine the perceptions and knowledge regarding their role in the etiology of head and neck cancers. It was a cross-sectional study, performed at Bilal colony in Karachi. Through systematic sampling, 425 subjects [a male and female from a household] were interviewed with a structured questionnaire. Knowledge regarding etiology of head and neck cancers was classified in ordinals of "good", "some" and "poor", for each substance separately, while practices were classified into "daily user", "occasional user" and "never user". About 40% of the participants were chewing at least one item [betel, areca or tobacco products] on daily basis. This prevalence was 2.46 times higher among males than females and 1.39 times higher among adolescents than adults. At least 79% of the participants were classified as having poor knowledge about the carcinogenicity of each of these items. Knowledge increased with age and level of education. Health hazards of these items were poorly recognized and about 20% perceived at least one of these items to be beneficial. Positive attitudes were seen regarding the steps to curb the production, business and consumption of these substances. In conclusion, prevalence of chewing of betel, areca and tobacco among adults and adolescents is high. Deficiency in knowledge and wrong perception of favorable effect of chewing products is common. Besides curtailing the availability of chewing products, correct knowledge regarding its ill-effects should be inculcated among population to decrease the burden of head and neck cancers.

Genetic diversity amongst landraces of a dioecious vegetatively propagated plant, betelvine (Piper betle L.).

Betelvine (Piper betle L., family Piperaceae) is an important, traditional and widely cultivated crop of India. The cultivators and consumers recognize more than 100 cultivars (landraces) based on regional and organoleptic considerations, while in terms of phytochemical constituents only five groups have been identified for all the landraces. Since betelvine is an obligate vegetatively propagated species, genomic changes, if any, may have become 'fixed' in the landraces. We carried out random amplified polymorphic DNA (RAPD) analysis in several landraces considered in four groups, namely, 'Kapoori', 'Bangla', 'Sanchi' and 'Others' in order to ascertain their genetic diversity. On the basis of the data from eleven RAPD primers, we distinguished genetic variation within and among the four groups of landraces. The results indicate the 'Kapoori' group is the most diverse. The neighbour joining (NJ) tree after a bootstrap (500 replicate) test of robustness clearly shows the four groups to be well separated. Interestingly, all known male or female betelvine landraces have separated in the NJ tree indicating an apparent gender-based distinction among the betelvines.

In Vitro Effect of Fungicides, Plant Extracts and Smoke on Conidial Germination of Fusarium oxysporum Root Rot Pathogen of Piper betle

Five fungicides such as rovral, bavistin, cupravit, dithane M-45 and thiovit were tested against conidial germination of Fusarium oxysporum. Dithane M-45 was the most effective against the fungus. Rests of the fungicides were more or less effective in the inhibition of conidial germination after 5~30 minutes immersion in 500~2500 ppm concentration. Five plant extracts(leaf extracts of Ocimum sanctum, Lantana camera, Calotropis procera, Azadirachta indica and Vinca rosea) found to be more or less effective against the fungus. 'Dhup' somke have good inhibitory effect on conidial germination of F. oxysporum.

Effect of ethanol extract of Piper betle Linn leaf on healing of NSAID-induced experimental ulcer--a novel role of free radical scavenging action.

Treatment with ethanol extract of leaf of P. betle at a dose of 150 mg/kg body weight daily for 10 days, after induction of peptic ulcer by NSAID in albino rats, produced significant healing effect. During healing process, on treatment with the extractive, antioxidative factor, e.g. superoxide dismutase and catalase activity, mucus and total gastric tissue sulfhydryl group were increased. In contrast, oxidised lipid and oxidatively modified proteins were reduced to near normalcy, within 7 to 10 days, however, change in the untreated group was not significant. The extract also showed significant in vitro free radical scavenging action. The results suggest that the antioxidant or free radical scavenging activity of the plant extract, may be responsible for its healing action.