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.
Piper betle ; Oryza--microbiology

Aims: Piper sarmentosum or locally known as Kaduk, is a tropical herb plant that was investigated for its phenolic content by previous researchers. The present study aimed at the analysis of crude methanolic extract of P. sarmentosum leaves for phenolic compounds identification and its anti-amoebic properties against pathogenic Acanthamoeba castellanii. Methodology and results: Folin-Ciocalteu assay was used to determine P. sarmentosum leaves methanolic extract (PSLME)’s total phenolic content (TPC). The extract was further characterized by using gas chromatography-mass spectrometry (GC-MS), reverse phase-high performance liquid chromatography (RP-HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses to determine the chemical constituents in methanolic PSLME extract. The cytotoxicity of the extract was evaluated through the determination of inhibition concentration for half of cell population (IC50) of pathogenic A. castellanii followed by cell morphological analysis using inverted light and scanning electron microscopies. Acridine-orange/Propidium iodide (AOPI) staining was also conducted to determine the integrity of cell membrane for quantitative analysis. The results demonstrated that the TPC from PSLME was 142.72 mg [GAE]/g with a total of 33 phenolic compounds identified. The IC50 value obtained for A. castellanii was low (74.64 μg/mL) which indicates promising anti-acanthamoebic activity. Microscopy analyses showed that the plant extract caused cells encystment, in which exhibited by distinctive morphological changes on the cells shape and organelle, as well as shortening of acanthopodia. The dual staining and its quantitative analysis prove compromised membrane integrity in the treated amoeba. Conclusion, significance and impact of study This finding provides the evidence that PSLME contains active phenolic compounds contributing to the anti-acanthamoebic activity on pathogenic Acanthamoeba species.
Piperaceae ; Acanthamoeba castellanii--pathogenicity

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.
Azadirachta ; Calotropis ; Fungi ; Fusarium* ; Germination* ; Immersion ; Lantana ; Ocimum ; Piper betle* ; Piper* ; Plant Extracts* ; Plants* ; Smoke* ; Vinca

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).
Piper betle ; Staphylococcus aureus ; Anti-Bacterial Agents ; Biofilms

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).
Piper betle ; Staphylococcus aureus ; Anti-Bacterial Agents ; Biofilms

Aims: Piper nigrum L. (black pepper) is an economically important commodity plant in Malaysia, which generated RM 200.95 million from pepper export in the year of 2018. However, the increase in pepper production is restricted by diseases. Fusarium wilt is one of the major diseases of P. nigrum L. The objectives for this study were to isolate Fusarium spp. associated with Fusarium wilt of P. nigrum L. from selected pepper farms in the northwestern region of Sarawak and to characterize the Fusarium spp. isolated morphologically and molecularly. Methodology and results: Fusarium spp. were isolated from diseased root samples. The pathogen was grown on potato dextrose agar (PDA) under dark condition at circa (ca.) 25 °C for morphological characterisation. Molecular characterisation was done by using internal transcribed spacer (ITS). Phylogenetic tree was constructed to study the genetic relationship of the isolates. Fusarium solani, F. oxysporum, F. proliferatum were the three Fusarium species identified. There were variations in morphological characters observed between and among the species, including the colony form, margin, elevation, surface appearance and pigmentation. No distinctive morphological characteristic was specific to a location. In addition, growth rate, macroconidia sporulation rate, and microconidia sporulation rate of the isolates were not correlated. In molecular phylogeny, the three Fusarium species were separated into three distinct clades representing the three identified species. The genetic relatedness between isolates within each species was depicted in the tree. Conclusion, significance and impact of study Variations were observed among isolates in this study based on morphological and molecular characterization. This study would contribute information on the variations of Fusarium spp. associated with Fusarium wilt of P. nigrum L. from the northwestern region of Sarawak.
Fusarium ; Fusariosis ; Piper nigrum

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.
Alkaloids ; analysis ; isolation & purification ; Chromatography, Thin Layer ; Lignans ; analysis ; isolation & purification ; Piper betle ; chemistry ; Plant Stems ; chemistry

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).
Antineoplastic Agents, Phytogenic/pharmacology* ; Cell Movement/drug effects* ; HT29 Cells ; Humans ; Microtubules/drug effects* ; Piper betle ; Plant Extracts/pharmacology* ; Plant Leaves

OBJECTIVE: The aqueous and pelletized admixture of Piper nigrum L. was evaluated for its oviposition response and larvicidal activity against Aedes aegypti mosquitoes.

METHODS: The aqueous and pelletized extract of Piper nigrum L. was prepared and first tested in the laboratory. Efficiency is evaluated using the mosquito-chamber test. A small-scale field test was also done to determine the oviposition response of the pepper extract to ovicidal-larvicidal (OL) traps. Larvicidal bioassay following the WHO standard protocols with slight modification at different concentrations was performed.

RESULTS: Results of the mosquito chamber test in the laboratory showed that the aqueous solution exhibited an increasing rate of oviposition attraction of female Aedes aegypti to increasing rate of concentration with an average of 70% attraction at 1000 ppm as compared to 30% attraction to OL traps with water alone. The aqueous pepper-based solution and pelletized pepper solution at 1000 and 2000 ppm are considered attractants to Aedes mosquitoes. Both solutions have oviposition activity index (OAI) of > +0.3. It was also field tested on the 10 buildings within the Department of Science and Technology (DOST) compound. Results showed an oviposition ratio two times better in both the 1000 ppm concentration of the aqueous pepper-based solution and pelletized pepper solution as compared to the control. The positive ovitrap index was in the range of 78%-84% for both the aqueous pepper-based and pelletized pepper against water which is 70.0%. Larvicidal activity of the aqueous pepper-based solution against 3rd larval instars of Aedes aegypti at increasing dosages from 75mg/1 to 600 mg/I had an LCso of 127 mg/I and 395 mg/I for LC90 The LCso for the solution with pelletized pepper at the same dosing concentration is 117 mg/I with LC90 of 285 mg/1. The results also showed that these can be used to control larval instars of the Aedes aegypti mosquitoes.

CONCLUSION: The overall results indicate that the aqueous and pelletized extracts of Piper nigrum L. are effective in attracting the Aedes aegypti mosquitoes for oviposition and exhibit a larvicidal activity.

Animal ; Aedes ; Piper Nigrum ; Larva ; Oviposition ; Water ; Biological Assay

OBJECTIVE: This study was conducted to validate the effectiveness of the ovicidal-larvicidal (OL) trap with the pelletized extracts of Piper nigrum L. in attracting female Aedes aegypti mosquitoes for oviposition and in reducing teh number of larvae hatched, thus preventing the emergence of adult mosquitoes.

METHODS: The OL trap system containing the developed plant-based pellets from Piper nigrum L. was field tested in Marikina and Quezon City. Paired OL traps were installed outdoors in 150 selected houses in each city. The OL trap is composed of a black painted can, a small strip of lawanit (paddle) for oviposition and extracts of Piper nigrum L. as the ovicide-larvicide solution or plain water. Pelletized form of Piper nigrum L. was prepared by grinding the seeds then mixing it with 1% previously cooked starch solution at a ratio of 1:1 followed by extrusion and forming into pellets using a granulator then drying. The number of positive OL traps was recorded. Larval mortality and the Ovitrap Index were also determined.

RESULTS: Four collections were made using the pelletized form in which 2,314 OL traps were collected. The OL traps with the pelletized form showed a 53.2% attraction which is significantly higher than that with water (p-value<0.0001). The percent egg and larval mortality in both cities was recorded at 86.2% while adult emergence is 13.8%. The Ovitrap Index in Marikina was in the range of 53% to 68% while OL traps with water have an Ovitrap Index range of 39% to 65%. In Quezon City, OL traps with the pelletized admixture have an Ovitrap Index range of 35% to 50% while those with water have a range of 31% to 36%.

CONCLUSION: The overall results indicate that the developed OL trap system with Piper nigrum L. is effective in attracting Aedes aegypti mosquitoes. The OL traps with extracts of Piper nigrum L. were also found to be lethal to larvae.

Animal ; Aedes ; Piper Nigrum ; Larva ; Oviposition ; Starch ; Water ; Cities ; Desiccation ; Seeds