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Item Screening Anti-Bacterial Activity of Endoi.Ichenic Fungi Collected From Negombo Lagoon Sri Lanka(Proceedmgs of the Postgraduate Institute of Science Research Congress. Sri Lanka, 2019) Weerasinghe, W.R.H.; Attanayake, R.N.; Weerakoon, G.; Paranagama, P.A.From the ancient past, natural products have been in use to fulfill a variety of human needs, from sustenance to pharmaceutical necessities. With the discovery of Endolichenic fungi (ELF) as a novel source prominent bioactiOt1es in recent years. a new trend has been created to explore more about these fascinating organisms, which live inside the lichen thallus asymptomatically. ELF isolated from the lichens collected from mangrove plants In Negombo lagoon's Kadol kale area (Lititude: 701 Longitude: were identitied using molecular techniques and were subjected to anti-bacterual assay. Ethyl acetate crude extracts of 18 identified endolichenic fungal strains were obtained and their anti-bacterial activities were tested against aerobic bactertal species Escherichia coli, Bacillus subtilis and Staphvlococcus aureus. The assay was carried out using agar well diffusion method using HI of the extracts (5 mg/ml) with Azithromycin as the positive control. The inhibition zone diameters (in cm) against E. coli were 1.2 for Phanerothaete chnsosporium. 1.2 for Xvlaria feejecnsis, ().9 for Gelasinospora seminuda, 0.9 for Chaetomium globosum and 1.5 for Curvularia lunata while Azithromycin as the positive control showed a diameter of 2.1. The inhibition zone diameters against B. subtilis were 1.0 for Cochliobolus sp., 1.0 for P. chnsosporium, 1.8 for X. feeyeensls, 1.7 for G. seminuda, 1 2 for C. globosum, 1.5 for Hvpoxvlon lividipigmentum, 2.0 for (Pennicillium sp), 2.2 for C. lunata, 0.9 for Lasiodiplodia theobromae and 2.4 for positive control. The inhibition zone diameters against S. aureus were 1.1 for P. chrvsosporium, 22 for X'. feejeensis, 1.0 for G. seminuda, 1.0 for Chaetomium globosum, 2.0 for (Pennicillium sp). 1.8 for Curvularia lunata and 2.2 for positive control. Out of the 18 fungal strains, 9 showed significant activity against the tested bacterial species while Endomelanconiopsis endophvtica, Neofusicoccum parvum. Ncofusicoccum occulatum. Hvpoxvlon anthochroum. Cvtospora xvlocarpi, Cerrena sp., Schi:ophvllum commune, Daldinia eschscholt:i, Nodulisporium sp. fungal strains did not show any inhibition. Further studies will be conducted to isolate compounds responsible for these inhibitions.Item A NEW ANTIOXIDANT ACTIVE COMPOUND FROM THE ENDOLICHENIC FUNGUS, PENICILLIUM CITRINUM INHABITING THE LICHEN, PARMOTREMA SP(International Journal of Pharmaceutical Sciences and Research, 2019) Wickramarachchi, S.A.; Samanthi, U.; Wijeratne, K.; Paranagama, P.A.Context: Endolichenic fungi are the microorganisms living inside the thalli of the lichen and found to be a new source of bioactive secondary metabolites. This is an underutilized source of bioactive compounds and has not been studied extensively. Genus, Parmotrema is one of the foliose lichens with largely rounded lobes and occurs on rocks of rain forests, sub-montane and montane forests in Sri Lanka. Aims: The endolichenic fungi available in Sri Lanka and the chemistry of their secondary metabolites have not been explored thoroughly. This study aims at exploring novel bioactive compounds available in Penicillium citrinum inhabiting Parmotrema sp. Methods and Material: The lichen, Parmotrema sp., was collected from Hakgala Botanical Garden in Sri Lanka and its fungal strains were isolated and identified by Genomic DNA sequencing. Compounds were first extracted to ethyl acetate and subsequently subjected to bioassay-guided fractionation to isolate the bioactive compounds. The DPPH assay was used to determine the antioxidant activity. The active compound/s was separated by column chromatography, and its structure was elucidated by IR, 1D and 2D-NMR, 13C-NMR and MS. Statistical Analysis Used: The IC50 of the crude extract was calculated using Probit analysis (MINITAB® Release 14.1. Minitab Inc. 2003 Statistical Software). Results: A bioactive compound, named PP-PC-03 together with PP-PC-01 and PP-PC-02 was isolated from P. citrinum. All three compounds showed antioxidant activity in the DPPH assay with IC50 values 159.6 ± 22.3, 120.1 ± 11.7 and 68.6 ± 4.3 μg/mL respectively. Conclusion: Based on spectral analysis the structure of PP-PC-03was determined as 10-Ethylidene-2,4,9-trimethoxy-10,10a-dihydro-7,11-dioxa-benzo[b] heptalene-6,12-dione and identified as a new compound. Comparatively PP-PC-02 and PP-PC-03 had moderate antioxidant activities while PP-PC-01 showed strong radical scavenging activity.Item 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.