International Research Symposium on Pure and Applied Sciences (IRSPAS)

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Author: search.filters.author.Paranagama, P.A.

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    Evaluation of anti-inflammatory and antioxidant properties of Genus Piper in Sri Lanka. International Research Symposium on Pure and Applied Sciences,
    (International Research Symposium on Pure and Applied Sciences, 2017 Faculty of Science, University of Kelaniya, Sri Lanka., 2017) Athapattu, R.S.K.; Jayarathna, S.P.N.C.; Paranagama, P.A.; Senanayake, S.P.
    In Sri Lanka, the genus Piper is represented by ten species including five wild and two cultivated species. Most of the species of the genus Piper are important as medicinal plants and used in various systems of medicine.This study aims to evaluate the bioactivity (anti-inflammatory and antioxidant properties) of the selected five Piper spp. (P. nigrum L., P. longum L., P. betle L., P. chuvya (Miq.) and P. sylvestre). Specimens of leaves and fruits were collected from their natural habitats and cultivations, and were subjected to the extraction of essential oils by steam distillation. Three replicates were done for each concentration and data were analyzed using the MINITAB 17 statistical package. Antioxidant activity was tested using 2,2-diphenyl-1- picrylhydrazyl (DPPH) free radical scavenging assay and ferric reducing antioxidant power assay. The isolated essential oils of Piper species exhibited remarkable DPPH free radical scavenging ability at different concentrations. The results revealed that the essential oil of P.betle exhibited the highest radical scavenging activity (92.8% at 1 mg/mL) and the value is higher than even of the standard, Butylated Hydroxy Toluene (BHT), (74.5% at 1 mg/mL). Further, essential oils of P. chuvya and P. longum have shown 91.8% and 63.1%inhibition percentages at 1 mg/mL. Essential oil isolated from P. betle has showed remarkable ferric reducing antioxidant power (mean absorbance, 1.89at 1 mg/mL) compared to the standard, BHT, (2.01at 1 mg/mL). Further, it was observed that its antioxidant activity increased with the increment of the concentration of essential oil (0.03-1 mg/mL). These results suggested the potential of using essential oils of leaves of P.betle, P. chuvya (1.31) and P. longum (1.28) as sources of natural antioxidants. This study also focused on evaluating anti-inflammatory activity by using Human Red Blood Cell (HRBC) membrane stabilizing method.Higher mean inhibition percentage was observed for the essential oil of P. betle leaves (79.6%at 1 mg/mL) compared to the standard (aspirin) and the activity significantly increased with higher concentrations (76.4% at 0.5 mg/mL and 79.6% at 1 mg/mL). Further, 62 % and 58.3 % of inhibitions percentages were shown by essential oils of P. chuvya and P. longum respectively. According to the findings of this study, bioactivity properties of the leaves and fruits of these Piper species can be used in future perspectives for developing novel pharmaceuticals.
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    Anti-oxidant activity of selected endo lichenic fungi (ELF) in mangrove ecosystem of Puttalam lagoon.
    (International Research Symposium on Pure and Applied Sciences, 2017 Faculty of Science, University of Kelaniya, Sri Lanka., 2017) Maduranga, H.A.K.; Attanayake, R.N.; Amarasinghe, M.D.; Weerakoon, G.; Paranagama, P.A.
    Natural products based drug development has become an attractive area of research since there are limited options available to treat certain non-infectious diseases such as diabetes. Among these natural products, it has been reported that secondary metabolites of endolichenic fungi (ELF), have the ability to produce promising bioactive compounds. The objectives of this research were to isolate and identify ELF inhabiting mangroves in Puttalam lagoon, Sri Lanka using classical and DNA barcoding approach and to determine anti-oxidant activity of their secondary metabolites. Lichen hosts were collected from Puttalam lagoon in two different sites near, Athathale and around the NARA institute. The ELF were isolated following a standard procedure: a small piece of the thallus was surface sterilized, cut into pieces and dried on sterilized filter papers and then placed on malt extract agar in Petri dishes and incubated at room temperature (28 ⁰C – 30 ⁰C ) . Once pure cultures were obtained, seven isolates were randomly selected for DNA extraction following standard procedures. Quality of DNA was checked by agarose gel electrophoresis. Fungal internal transcribed spacer (ITS) region was amplified using polymerase chain reaction (PCR) with universal ITS 1 and ITS 4 primers and PCR products were sequenced using Sanger dideoxy chain-termination technology. DNA sequences were edited using BioEdit software and compared with the available sequences in the GenBank using Basic Local Sequence Alignment Search Tool (BLAST). In addition, morphological characterization of each fungal isolate was also carried out. Secondary metabolites from each isolate were extracted with ethylacetate separately and the solvent was evaporated under reduced pressure to obtain the crude extract. Free radical scavenging activity of the extracts were evaluated using 2, 2-diphenyl-1-picrylhydrdrazyl (DPPH) assay. Based on the highest sequence similarity to the GenBank sequences, isolates were identified as Diaporthe arengae (98 %), Neurospora crassa (100%), Lasiodiplodia theobromae (100 %), Schizophyllum commune (98 %), Diaporthe musigena (98 %), Hypoxylon anthochroum (98 %) and Nigrospora sphaerica (98%). IC50 values of extracts of Diaporthe arengae, Neurospora crassa and Lasiodiplodia theobromae were 375.9± 0.062μg/mL, 304.9±0.057 μg/mL and 211.2± 0.086 μg/mL respectively. Since percent inhibitions of the rest of the isolates were less than 50 % in the test doses, IC50 values were not calculated. All of the values were compared with standard Butylated Hydroxy Toluene (BHT) (IC50=108.0±0.072). Out of the seven ELF tested, L. theobromae showed the highest DPPH radical scavenging activity. Further testing of the rest of the isolates are being carried out and ELF may provide a good source of antioxidants for biotechnological applications.