0.1 mL of the PBE solutions of different concentrations The PBE of various concentrations were tested for their effects on the growth Were prepared as follows: The PBE was dissolved in deionized distilled waterĪt different concentrations (1000, 2000, 3000, 40 μg mL -1). To culture tubes that contained 15 mL of the culture medium. When the stationary growth phase was reached after 16 h. A 0.1 mL sample was taken from the culture Briefly, Pseudomonas aeruginosa PAO1 was grown Of PBE was assessed using a broth dilution method ( SmullenĮt al., 2007). The extracts were dissolved by sonicating the microfuge vials in a sonicator (DAIHAN) and stored at 4☌ until use.ĭetermination of MIC of PBE: The Minimum Inhibitory Concentration (MIC) The PBE (pH 6.5) was completely soluble in water. The extracts were weighed into sterile micro fuge vials and prepared into stocks of 20 mg mL -1 using sterile distilled water as diluents and sterilized by a 0.2 μm membrane filter. It was concentrated using a speed-vacuum concentrator. The supernatant was divided, into 1 mL aliquots, in micro fuge tubes. Further, the extract was centrifuged at 10,000 rpm to remove sediments. 100 g of pieces were boiled in 1 liter of deionized distilled water for many hours until the final volume was 100 mL. Dried leaves were shredded into small pieces. Fresh healthy leaves were washed with distilled water and air dried.
leaves were obtained from Mentakab, Pahang, Malaysia. Preparation of plant extract: Piper betle L. ML -1 by a spectrophotometer (Shimadzu) at 570 nm ( Nalina Cells concentration was standardized at 10 6 cells Were collected via centrifugation at 10000 rpm for 10 min and cells were resuspended Theīacterial specie was revived in nutrient broth at 37☌ overnight. Preparation of bacterial suspension: The stock of the PseudomonasĪeruginosa PA 01 was kept in glycerol at -70☌ for further use. Long and 0.3-0.6 μm wide) and can be cultured from almost all natural waters. This species is gram negative rod shaped (1.5-2 μm Its rapid reproduction rate and its significance as a pathogen ( LeeĮt al., 2010). PAO1 was used as a model bacterium because of its ability to foul surfaces rapidly, This work, a single representative bacterium, namely Pseudomonas aeruginosa To an extremely complex biofilm system that is poorly known. Model bacterial strain: Natural environment bacterial variance leads All materials were autoclaved for 20 min at 120☌ before use. A sterile 96 well clear flat bottom tissue culture microtiter plate (Corning, SIGMA) with a lid was used for biofilm assay. Glacial acetic acid (DMF, Fisher scientific) was used to resolubalize the dye bound to the adherent cells. Methanol (PROLABO) was used to fix the attached bacteria. Materials: The materials used in this study were nutrient broth for cell culture crystal violet dye (MERCK, 101408-0025) was used to stain the biofilm cells. Consequently, the present study was undertaken toĮvaluate the effects of Piper betle extract (PBE) on biofilm formationĪnd Extracellular Polymeric Substances (EPS) production by Pseudomonas aeruginosa.
Hypothesized that these may help to reduce biofilm formation ( SendamangalamĮt al., 2011). ( Kubo et al., 2006) and in this study, it was Many natural products of plants are well known for antimicrobial activities The cells using antibiotics, as practiced in industry, for example, does notĪlways work, because it is not usually possible to kill all the cells completelyįor an extended time and some cells still can attach onto the solid surface It would be better to preventīiofilm formation rather than killing the cells after it forms. That biological control of microbial attachment would be a novel promising alternativeįor mitigating membrane biofouling and would be a new niche that deserves further Sometimes it is hard to reach all the areas that are contaminated with biofilm.Īcidic and alkaline solutions are sometimes used to remove biofilm from surfacesīy washing but there is an issue of adverse environmental impact. ( RameshĮt al., 2006) and they may not be effective and energy efficient. Many physico-chemical methods haveīeen used, for regular physical and chemical cleaning, etc. To prevent or reduce membrane biofouling. So far, extensive research has been pursued to investigate the possible methods The control of membrane irreversibleįouling resulting from strongly bound fouling materials is a difficult and challenging The membrane life, increases the membrane cost and eventually adds additionalĬapital cost for membrane replacement.
High-quality purification systems have faced a major problem due to biofilmįormation on the membrane surface or biofouling. In wastewater treatment ( Yun et al., 2006). Membrane bioreactors (MBRs) have emerged as one of the innovative technologies