Effects of Piper betle Leaf Extract on Biofilm and Rhamnolipid Formation of Pseudomonas aeruginosa

Author(s):  
Irene Ratridewi ◽  
Shod A. Dzulkarnain ◽  
Andreas B. Wijaya ◽  
John T. R. Huwae ◽  
Daniel S. M. Putra ◽  
...  

High mortality rate and antimicrobial resistance are still becoming world-wide problems, due to Pseudomonas aeruginosa’s (P. aeruginosa) virulence and its ability to form biofilm. Biofilm’s formation is affected by the presence of rhamnolipid, whose production is regulated by quorum sensing systems. Piper betle (P. betle) possesses antimicrobial, antioxidant, anti-inflammatory and immunomodulatory properties. The aim of our study is to investigate the effects of P. betle leaf’s extract against biofilm formation and rhamnolipid production of P. aeruginosa. Active compounds of P. betle were identified using plate chromatography. Agar dilution method was used to determine the minimum biofilm inhibitory concentration (MBIC) of methanolic leaf extract of P. betle. A biofilm-producing P. aeruginosa isolate in the polystyrene plate adherence test was selected for confirmation of biofilm production by Scanning Electron Microscopy (SEM), after P. betle administration. Rhamnolipid detection and evaluation were performed by interpreting halo formed around the well. After administration of various concentrations of P. betle leaf extract on the microplate well, it was concluded that the MBIC of P. betle leaf extract on P. aeruginosa was 0.4%. Methanolic extract of P. betle leaf extract at concentration of 0.4% showed that P. aeruginosa could not form biofilm at all, although the bacteria could still aggregate and form a matrix. After linear regression analysis, beta-coefficient was obtained at -0.931 for P. betle leaf extract. It can be concluded that P. betle leaf extract was effective in inhibiting the growth of biofilm and formation of rhamnolipid by P. aeruginosa. The increase in concentration of P. betle leaf extract was inversely proportional to the diameter of the halo rhamnolipid formed. The higher the level of P. betel leaf extract, the smaller the diameter of the halo rhamnolipid formed.

2020 ◽  
Vol 8 (A) ◽  
pp. 928-931
Author(s):  
Irene Ratridewi ◽  
Shod Abdurrachman Dzulkarnain ◽  
Andreas Budi Wijaya ◽  
Wisnu Barlianto ◽  
Sanarto Santoso ◽  
...  

  BACKGROUND: Piper betle (P. betle) is widely used as a traditional medicine in Indonesia, with anti-quorum sensing and anti-biofilm activity. We investigated the impact of methanolic leaf extract of P. betle against Pseudomonas aeruginosa’s (P. aeruginosa) virulence factor, which associated with rhamnolipid (rhl) genes, METHODS: Minimum biofilm inhibitory concentration of the extract was determined at a concentration of 0.4% by agar dilution assay. The expression of rhlA and rhlC gene was assessed by using real-time polymerase chain reaction. RESULTS: All P. aeruginosa isolates contained rhlA, rhlB, and rhlC genes, which associated with rhl production. The expression of the rhlC gene decreased after administration of P. betle leaf extract at concentration of 0.4%, with beta coefficient was 0.662 (p = 0.019). CONCLUSION: The methanolic leaf extract of P. betle shows inhibition of rhlC gene expression, indicating the anti-rhl properties of P. betle against P. aeruginosa infection.


2020 ◽  
Vol 8 (2) ◽  
Author(s):  
Gwendolyn L. Kapondo ◽  
. Fatimawali ◽  
Meilani Jayanti

Abstract: Betel leaf (Piper betle L) contains alkaloid compounds and has antibacterial and antiseptic functions. The purpose of this research was to isolated alkaloid compounds also to tested the effectiveness of inhibition of the betel leaf extract against the Staphylococcus epidermidis bacterial growth. Simplicia betel leaf was extracted used 96% of ethanol solvent. Antibacterial activity tested of betel leaf extract with a concentration variation of 25%, 20%, 15%, 10% and 5% using the liquid dilution method. The results of identification of alkaloid isolates used UV-Vis Spectrophotometry method can be known alkaloid compounds contained in betel leaves include alkaloids with indole base framework that was absorption at 262 nm and 274 nm wavelengths. The results of the effectiveness of betel leaf on Bacteria Staphylococcus epidermidis used the liquid dilution method showed that betel leaf extract with Optical density values at concentrations of 25%, 20%, 15% and 10% before and after incubation decreased respectively by -0.347, -0.304, -0.192 and -0.104, while at a concentration of 5% there was increase in Optical density values of 0.162. From the results of the research, it can be concluded that the ethanol extract of betel leaf contains an alkaloid with an indole base framework and was inhibitory activity against Staphylococcus epidermidis bacteria with a MIC at a concentration of 10%.Keywords: betel leaf, Staphylococcus epidermidis, alkaloid compound, liquid dilution.  Abstrak: Daun Sirih (Piper betle L) mengandung senyawa alkaloid dan memiliki fungsi sebagai antibakteri dan antiseptik. Penelitian ini bertujuan untuk mengisolasi senyawa alkaloid dan menguji efektivitas penghambatan dari ekstrak daun sirih hijau terhadap pertumbuhan bakteri Staphylococcus epidermidis. Simplisia daun sirih hijau diekstraksi dengan menggunakan pelarut etanol 96%. Uji aktivitas antibakteri ekstrak daun sirih dengan variasi konsentrasi 25%, 20%, 15%, 10% dan 5% menggunakan metode dilusi cair. Hasil identifikasi terhadap isolat alkaloid menggunakan metode Spektrofotometri UV-Vis dapat diketahui senyawa alkaloid yang terkandung dalam daun sirih termasuk alkaloid dengan kerangka dasar Indol yang mempunyai serapan pada panjang gelombang 262 nm dan 274 nm. Hasil uji efektivitas daun sirih terhadap bakteri Staphylococcus epidermidis dengan metode dilusi cair menunjukan bahwa ekstrak daun sirih dengan nilai densitas optik pada konsentrasi 25%, 20%, 15% dan 10% sebelum dan sesudah inkubasi mengalami penurunan berturut-turut sebesar -0.347, -0.304, -0.192 dan -0.104, sedangkan pada konsentrasi 5% mengalami kenaikan nilai densitas optik sebesar 0.162. Dari hasil penelitian yang diperoleh, dapat disimpulkan bahwa ekstrak etanol daun sirih mengandung alkaloid dengan kerangka dasar indol dan memiliki aktivitas penghambatan terhadap bakteri Staphylococcus epidermidis dengan KHM pada konsentrasi 10%.Kata Kunci: daun sirih, Staphylococcus epidermidis, senyawa alkaloid, dilusi cair.


2019 ◽  
Vol 10 (2) ◽  
pp. 44-50
Author(s):  
Rinaldi Daswito ◽  
Rima Folentia ◽  
M Yusuf MF

One of the diseases that can be transmitted by flies is diarrhea. Green betel leaf contains essential oils, chavicol, arecoline, phenol, and tannins which function as plant-based insecticides. This study aimed to determine the effectiveness of green betel leaf extract (Piper betel) as a plant-based insecticide on the number of mortality of house flies (Musca domestica). The research was an experimental study used After Only Design used the One Way Anova test with a 95% confidence level. The samples used were 360 ​​house flies. Each treatment of 30 house flies with 4 repetitions and used three concentrations of green betel leaf extract (25%, 50%, 75%). The study was conducted at the Chemistry and Microbiology Laboratory of Health Polytechnic Tanjungpinang, while the location of the fly collection was at the Tokojo Garbage Collection Station in Bintan Regency. The number of mortality of house flies at a concentration of 25% was 81 heads (67.5%), 50% concentrations were 93 heads (77.5%), and at a concentration of 75% were 103 heads (85.83%). There was an effect of green betel leaf extract on the mortality of house flies (p-value 0.0001 <0.05) with the most effective concentration of 75%. Further research is needed to obtain a finished product utilizing green betel leaf extract as a vegetable insecticide, especially in controlling the fly vector. Need further research on the use of green betel leaf extract as a vegetable insecticide controlling the fly vector by taking into account the amount of spraying and the age of the fly.   Keywords: Green betel leaf extract , organic insecticide, houseflies


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 295
Author(s):  
Sebastián Candelaria-Dueñas ◽  
Rocío Serrano-Parrales ◽  
Marisol Ávila-Romero ◽  
Samuel Meraz-Martínez ◽  
Julieta Orozco-Martínez ◽  
...  

In Tehuacán-Cuicatlán valley (Mexico), studies have been carried out on the essential oils of medicinal plants with antimicrobial activity and it was found that they present compounds in common such as: α-pinene, β-pinene, carvacrol, eugenol, limonene, myrcene, ocimene, cineole, methyl salicylate, farnesene, and thymol. The goal of this study was to assess the antimicrobial activity of essential oils’ compounds. The qualitative evaluation was carried out by the Kirby Baüer agar diffusion technique in Gram-positive bacteria (11 strains), Gram-negative bacteria (18 strains), and yeasts (8 strains). For the determination of the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), the agar dilution method was used. All the evaluated compounds presented antimicrobial activity. The compounds eugenol and carvacrol showed the largest inhibition zones. Regarding yeasts, the compounds ocimene, cineole, and farnesene did not show any activity. The compounds eugenol, carvacrol, and thymol presented the lowest MIC; bactericidal effect was observed at MIC level for S. aureus 75MR, E. coli 128 MR, and C albicans CUSI, for different compounds, eugenol, carvacrol, and thymol. Finally, this study shows that the essential oils of plants used by the population of Tehuacán-Cuicatlán valley share compounds and some of them have antibacterial and fungicidal activity.


1992 ◽  
Vol 108 (3) ◽  
pp. 449-455 ◽  
Author(s):  
G. Tzanakaki ◽  
C. C. Blackwell ◽  
J. Kremastinou ◽  
C. Kallergi ◽  
G. Kouppari ◽  
...  

SUMMARYUsage of antibiotics in southern Europe is less well regulated than in northern countries. The proportion (48%) of meningococci in Spain insensitive to penicillin (MIC ≥ 0·1 mg/l) prompted this investigation of antibiotic sensitivities of isolates from Greek patients with meningitis (31) and carriers (47 school-children and 472 recruits). The agar dilution method was used to determine MIC to penicillin G (PN), sulphamethoxazole (SU), rifampicin (RF), cefaclor (CF) and ciprofloxacin (CP).The proportion of isolates insensitive to PN was 48% for isolates from patients, 19% from school-children and 36·6% from recruits. Resistance to SU (MIC ≥ 16 mg/l) was found in 16% of those from patients, 10·6% from children and 40% from recruits. None of the isolates from patients was resistant to RF (≥ 1 mg/l) but 6% of those from carriers were. Resistance to CF (≥ 4 mg/l) was found in 9·2% of patient isolates, 6·4% from children and 23·7% from recruits. All isolates except one were sensitive to CP (MIC range < 0·0015–0·125 mg/l).Resistances to PN, SU and RF were analysed by serogroup, serotype and subtype of the bacteria. The proportion of resistant isolates showed some variation between different areas of Greece, but it was not statistically significant.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Zhao-Yu Guo ◽  
Zhuo-Yang Zhang ◽  
Jia-Qi Xiao ◽  
Jin-Hong Qin ◽  
Wei Zhao

Aim. The study was conducted to investigate the antibacterial and antiasthmatic effects of Nandina domestica leaf extract, to find out its active components, and to assess its safety issue. Methods. (1) Solid-phase agar dilution method was used for antibacterial activity test of nandina leaf extract and the change of bacterial morphology after treatment was observed under the transmission microscope; (2) guinea pig model of asthma was used to test the asthma prevention effect of nandina leaf extract; (3) alkaloids and flavones were separated from nandina leaf extract and were further analyzed with HPLC-MS; (4) mice model was used to assessment of the safety issue of nandina leaf extract. Results. (1) Nandina leaf extract inhibited the growth of bacteria and destroyed bacterial membrane; (2) nandina leaf extract alleviated animal allergy and asthma; (3) the components reextracted by ethyl acetate were active, in which alkaloids inhibited Gram-positive bacteria and prevented asthma and flavones inhibited Gram-negative bacteria; (4) nandina leaf extract had no toxic effect on mice. Conclusion. Nandina leaves inhibit bacterial growth and prevent asthma through alkaloids and flavones, which had integrated function against chronic bronchitis. This study provided theoretical basement for producing new Chinese medicine against chronic bronchitis.


2013 ◽  
Vol 7 (11) ◽  
pp. 804-811 ◽  
Author(s):  
Sabrina Nedjai ◽  
Abouddihaj Barguigua ◽  
Nassima Djahmi ◽  
Loubna Jamali ◽  
Khalid Zerouali ◽  
...  

Introduction: Expended spectrum β-lactamase (ESBL)-producing Enterobacter cloacae is an important nosocomial pathogen. In this study, the prevalence and the molecular epidemiology of ESBL producing E. cloacae strains isolated from various hospitals in Annaba, Algeria were investigated. Methodology: The study involved 63 isolates of E. cloacae obtained during 2009 at the four hospitals in Annaba. The detection of ESBL was performed using the double-disk synergy test and the combined disk test. Minimum inhibitory concentrations (MICs) were determined using the agar dilution method. The presence of blaCTX-M, blaSHV, blaTEM, and blaDHA β-lactamase genes was evaluated by PCR, and genomic typing was determined by pulsed-field gel electrophoresis (PFGE) analysis. The clinical and microbiological data were entered into the EpiI Info database. Results: Thirty isolates (47.6%) had an ESBL phenotype. BlaCTX-M group1 (76%); blaTEM (70%) were the most prevalent, followed by blaDHA (16.6%) and blaSHV (10%). Eighteen strains expressed at least two bla genes. MICs revealed a high level of resistance to cefotaxime, ceftazidime, and cefepime.  PFGE revealed an epidemic clonal dissemination of these isolates. Various risk factors associated with the occurrence of ESBL-producing E. cloacae were detected. Conclusions: A higher frequency of ESBL-producing isolates and a diversity of β-lactamases were detected among ESBL-producing E. cloacae; these resulted from an epidemic clonal dissemination and high transference of ESBL genes between bacteria in hospital settings. Strict measures will be required to control the further spread of these pathogens in hospital settings.


2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Séverine Boisard ◽  
Anne-Marie Le Ray ◽  
Anne Landreau ◽  
Marie Kempf ◽  
Viviane Cassisa ◽  
...  

During this study, thein vitroantifungal and antibacterial activities of different extracts (aqueous and organic) obtained from a French propolis batch were evaluated. Antifungal activity was evaluated by broth microdilution on three pathogenic strains:Candida albicans, C. glabrata, andAspergillus fumigatus. Antibacterial activity was assayed using agar dilution method on 36 Gram-negative and Gram-positive strains includingStaphylococcus aureus. Organic extracts showed a significant antifungal activity againstC. albicansandC. glabrata(MIC80between 16 and 31 µg/mL) but only a weak activity towardsA. fumigatus(MIC80= 250 µg/mL). DCM based extracts exhibited a selective Gram-positive antibacterial activity, especially againstS. aureus(SA) and several of its methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains (MIC10030–97 µg/mL). A new and active derivative of catechin was also identified whereas a synergistic antimicrobial effect was noticed during this study.


2014 ◽  
Vol 44 (2) ◽  
pp. 287-290 ◽  
Author(s):  
Carolinie Batista Nobre da Cruz ◽  
Fabio Alessandro Pieri ◽  
Gislene Almeida Carvalho-Zilse ◽  
Patrícia Puccinelli Orlandi ◽  
Carlos Gustavo Nunes-Silva ◽  
...  

Honeys are described possessing different properties including antimicrobial. Many studies have presented this activity of honeys produced by Apis mellifera bees, however studies including activities of stingless bees honeys are scarce. The aim of this study was to compare the antimicrobial activity of honeys collected in the Amazonas State from Melipona compressipes, Melipona seminigra and Apis mellifera against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, and Candida albicans. Minimum inhibitory concentrations were determined using the agar dilution method with Müller-Hinton agar (for bacteria) or Saboraud agar (for yeast). Staphylococcus aureus and E. faecalis were inhibited by all honeys at concentrations below 12%, while E. coli and C. violaceum were inhibited by stingless bee honeys at concentrations between 10 and 20%. A. mellifera honey inhibited E. coli at a concentration of 7% and Candida violaceum at 0.7%. C. albicans were inhibited only with honey concentrations between 30 and 40%. All examined honey had antimicrobial activity against the tested pathogens, thus serving as potential antimicrobial agents for several therapeutic approaches.


Sign in / Sign up

Export Citation Format

Share Document