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Item Unveiling the Probiotic Properties of Lactic Acid Bacteria Inhabiting Tender Coconut Water(, Centre for Research and Knowledge Dissemination (CRKD) of the Sabaragamuwa University of Sri Lanka, 2023) Pavalakumar, D.; Undugoda, L.J.S.; Manage, P.M.; Nugara, R.N.; Kannangara, S.This research delves into the unexplored potential of tender coconut water as a rich source of probiotics, with a focus on identifying promising probiotic lactic acid bacteria (LAB). By employing biochemical screening techniques, four potential LAB strains were isolated: Lactiplantibacillus plantarum CWJ3, Lacticaseibacillus paracasei CWKu14, Lacticaseibacillus rhamnosus CWKu12, and Lacticaseibacillus casei CWM15. These strains were comprehensively assessed to evaluate their resilience under various conditions: pH (2, 3, and 6.5), bile (0.3% and 0.5%), phenol (0.4% and 0.6%), salt (3% and 6%) concentrations, and temperatures (20°C, 37°C, 45°C, and 60°C). Further, resistance to lysozyme (100 mg L-1), artificial saliva juice (0.3% α-amylase, at pH 6.9), simulated gastric juice (0.3% pepsin, at pH 2), and simulated intestinal juice (0.3% Ox-gall and 0.1% pancreatin at pH 7) were determined. Their antibiotic susceptibility was tested against ten antibiotics at 100 ppm, and cell surface hydrophobicity, auto-aggregation, co-aggregation with selected pathogens, antibacterial activity, hemolytic activity, and DNase activity, also were evaluated. The statistical analysis demonstrated the survival of all four strains even in demanding circumstances, with Lactiplantibacillus plantarum CWJ3 displaying the greatest resilience. Optimum growth occurred at 37°C, while none of the strains survived at 60°C. A pH of 6.5 was optimal for their growth, and Lactiplantibacillus plantarum CWJ3 stood out as the only strain capable of withstanding pH 2 after 2 hours. Lacticaseibacillus rhamnosus CWKu12 and Lacticaseibacillus casei CWM15 showed the highest percentage of auto-aggregation and co-aggregation while Lacticaseibacillus rhamnosus CWKu12 and Lactiplantibacillus plantarum CWJ3 exhibited high cell surface hydrophobicity. The strains displayed susceptibility to Clarithromycin, Erythromycin, and Azithromycin, but resistance to Cefuroxime and Streptomycin antibiotics. All four strains inhibited various food-borne pathogens, showcasing their potential as probiotics. Moreover, they displayed no hemolytic or DNase activity. Remarkably, Lactiplantibacillus plantarum CWJ3 excelled in challenging environments, making it a noteworthy contender for a probiotic role.Item Assessing the antioxidative potential of lactic acid bacteria inhabiting tender coconut water(Faculty of Graduate Studies - University of Kelaniya, Sri Lanka, 2023) Pavalakumar, D.; Undugoda, L.J.S.; Manage, P.M.; Nugara, R.N.; Kannangara, S.Probiotics are renowned for their beneficial impact on human health, including their antioxidative properties. Lactic acid bacteria (LAB), which constitute a major group within probiotics, have been recognized for their ability to act as potent antioxidants, providing relief from oxidative stress in the host. This study aims to evaluate the antioxidative activity of probiotic LAB inhabiting the tender coconut water samples collected from different regions of Sri Lanka. In a previous study, analysis of the collected tender coconut samples resulted in the identification of four potential probiotic strains at the species level: Lactiplantibacillus plantarum CWJ3, Lacticaseibacillus rhamnosus CWKu12, Lacticaseibacillus paracasei CWKu14, and Lacticaseibacillus casei CWM15. The antioxidative activities of these strains were evaluated using four different chemical antioxidant assays: the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay at 517 nm, the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay at 734 nm, the ferric reducing antioxidant power (FRAP) assay at 593 nm, and the total phenolic content (TPC) assay at 765 nm through spectrophotometric method. Three different concentrations of each bacterial strain (107, 108, and 109 CFU mL-1) were prepared using overnight grown cultures in deMan, Rogosa, and Sharpe broth at 37 °C, included in the study. The results indicate a significant increase (p < 0.05) in antioxidative activity for all four strains across all analyzed methods with an increase in cell concentration. Lactiplantibacillus plantarum CWJ3 exhibited the highest antioxidative potential at 109 CFU mL-1, with 88.85±0.84% radical scavenging activity in the DPPH assay, 84.0±0.14% radical scavenging activity in the ABTS assay, 0.029±0.004 mg mL-1 ascorbic acid equivalent in the FRAP assay, and 0.0052±0.0006 mg mL-1 gallic acid equivalent in the TPC assay. Conversely, Lacticaseibacillus paracasei CWKu14 demonstrated the lowest values for all antioxidative activities. Continued research on the antioxidative properties of probiotic LAB derived from tender coconut water holds promising prospects for the development of functional beverages.Item Biocontrol potential of endophytic fungi in tea (Camellia sinensis (L.) Kuntze) leaves against selected fungal phytopathogens(Malaysian Journal of Microbiology, 2022) Thambugala, K.; Daranagama, D.; Kannangara, S.Aims: Endophytic fungi are a diverse group of microorganisms that stay asymptomatically in the healthy tissues of the host. Many fungal endophytes are associated with the tea plant (Camellia sinensis) and the pathogens of the tea plant have the potential to grow as endophytes or act as latent pathogens during the initial growth of the plant. The present study aimed at screening tea endophytic fungi with the potential for control of a few critical phytopathogens, Fusarium sp., Lasiodiplodia theobromae, Pestalotiopsis sp. and Sclerotinia sclerotiorum while evaluating the efficiency of growth inhibition of these phytopathogens by the endophytic fungal isolates using in vitro assays. Methodology and results: Five endophytic fungal strains; Colletotrichum gloeosporioides, C. siamense, Daldinia eschscholtzii, Pseudopestalotiopsis chinensis and Phyllosticta capitalensis isolated from leaves of C. sinensis in Sri Lanka were evaluated for growth inhibition against plant pathogens; Fusarium sp., L. theobromae, Pestalotiopsis sp. and S. sclerotiorum using the dual culture assay and volatile compound-mediated inhibition assay. All the fungal endophytes used in this study exhibited antifungal activity against Fusarium sp., Pestalotiopsis sp. and S. sclerotiorum in the dual culture assay on PDA. Daldinia eschscholtzii (67.30%) and C. gloeosporioides (61.54%) showed strong antagonistic activity against S. sclerotiorum, while P. capitalensis (42.30%) demonstrated moderate activity. All the endophytic fungal strains showed moderate antifungal activities against Fusarium sp. The percentage growth inhibitions of Pestalotiopsis sp. by all the endophytic isolates tested were below 25.00%. In the volatile compound-mediated inhibition assay, none of the endophytic isolates showed visible inhibition against the phytopathogens used. Conclusion, significance and impact of study: In this study, the fungal endophytes which showed potential antagonistic activity against the tested phytopathogens can be used to develop commercialized products of fungal biocontrol agents (BCAs) for controlling plant fungal diseases.Item Suppression and management of Meloidogyne incognita in soil using Trichoderma harzianum NFCF160 and Trichoderma virens Isf-77(Malaysian Journal of Microbiology, 2022) Rajakaruna, N.; Undugoda, L.; Kannangara, S.; Abeywickrama, K.Aims: Meloidogyne incognita adversely affects numerous crop plants worldwide. Therefore, the modern world has been moving towards biocontrol methods to prevent nematode attacks. This study was aimed to (i) investigate the potential use of Trichoderma harzianum NFCF160 and T. virens Isf-77 in managing M. incognita in soil and (ii) identify trapping mechanisms employed by both Trichoderma strains to suppress M. incognita. Methodology and results: Three weeks old, Basella alba L. plants were subjected to five different treatments. The above and below ground growth parameters and the galling indices of these plants were measured every four weeks for three sampling times. Trapping mechanisms employed by Trichoderma strains were examined following plate assays. Plants treated with T. harzianum NFCF160 and T. virens Isf-77 had significantly higher values for the total number of leaves (34 ± 2.84) and (27 ± 2.61), fresh weight of the shoot (81 ± 9.51 g) and (91 ± 9.70 g), dry weight of the shoot (71 ± 5.24 g) and (62 ± 5.81 g), respectively eight weeks after inoculation of M. incognita. Significantly low galling indices (2 and 2) were recorded in B. alba treated with Trichoderma strains. Both Trichoderma strains exhibited various nematode-trapping mechanisms, such as non-constricting rings and adhesive spores. Conclusion, significance and impact of study: This investigation highlighted the potential of both Trichoderma strains as biocontrol agents to control M. incognita effect in sustainable agriculture.Item Depolymerization of polyaromatic hydrocarbons by Penicillium spp. inhabit the phyllosphere of urban ornamental plants(Environmental Quality Management, 2022) Dharmasiri, R. B. N.; Undugoda, L. J. S.; Nilmini, A. H. L.; Pathmalal, M. M.; Nugara, N. N. R. N.; Udayanga, D.; Kannangara, S.A variety of anthropogenic sources release hazardous polyaromatic hydrocarbons (PAHs) into the phyllosphere which is an excellent niche for diverse fungi, and some of them have PAHs degradation capabilities. Therefore, this research attempted to determine the PAHs (phenanthrene, anthracene, naphthalene, and pyrene) degradation capability of phyllosphere inhabited Penicillium species. The leaf samples were collected from highly polluted urban areas (Panchikawatta, Pettah, Orugodawatta, Maradana, Sapugaskanda, and Colombo Fort) in Sri Lanka to isolate fungal species inhabiting the phyllosphere. Furthermore, their distribution patterns among the leaf tissue layers were studied using bright-field microscopic observations. Moreover, the best PAHdegraders were screened out using plate assays and confirmed throughHigh Performance Liquid Chromatography (HPLC) analysis. Further, their enzymatic activities during the PAHs degradation were analyzed. As per the microscopic observations, the highest fungal distribution was in the upper epidermis of the leaves followed by the fungal distribution in the interspaces of palisade mesophyll layers. Out of isolated fungal species, two Penicillium spp. (Penicillium citrinum P23B-91 and Penicillium griseofulvum P9B - 30) showed the highest PAHs (phenanthrene, anthracene, naphthalene, and pyrene) degradation capabilities. Manganese peroxidase (MnP) enzyme dominated phenanthrene degradation in P. griseofulvum P9B - 30, which showed the highest phenanthrene degradation ability (61%). In addition, P. citrinum P23B-91 was good at degrading anthracene (88%) and also displayed a higher MnP activity during the anthracene degradation than laccase and lignin peroxidase activities. The discoveries from the toxicity assay during the PAHs degradation processes revealed that the produced byproducts had no toxic effects on the fungal growth cycle and the phyllosphere. Therefore this phyllosphere Penicillium spp. are ideal for the bioremediation of polluted air in urbanized areas.Item Fungal pretreatment to enhance the yield ofphytochemicals and evaluation of α-amylase and α-glucosidase inhibition using Cinnamomum zeylanicum (L.)quills pressurized water extracts(Letters in Applied Microbiology, 2020) Wariyapperuma, W.A.N. Madushika; Kannangara, S.; Wijayasinghe, Y.S.; Subramanium, S.; Jayawardena, B.Bioactive compounds entrapped in plant materials can be effectively recovered using fungal enzymes. Cinnamomum zeylanicum Sri Wijaya (SW) and Sri Gemunu (SG) accessions and commercially available C. zeylanicum (CC) were subjected to fungal pretreatment and extracted with pressured water (PWE, 0 098 MPa). Thirteen fungal species were isolated and the substrate utilization ability of the species was tested using cellulose, pectin and lignin (indirectly). Total phenolic content (TPC, Folin–Ciocalteu method), proanthocyanidin content (PC, vanillin method) and α-amylase and α-glucosidase inhibitory potential of the extracts were evaluated. The anti-diabetic drug, Acarbose was used as the positive control. Trichoderma harzianum (MH298760) showed the highest cell lysis ability and hence was used for the microbial pretreatment process. Extracts of SW treated with T. harzianum species (Pre-SW) gave the highest percentage yield (4 08% 0 15%), significantly potent inhibition (P < 0 05) of α-amylase and α-glucosidase activities (IC50 57 8 and 36 8 μg ml−1 respectively), TPC (2 24 0 02 mg gallic acid equivalent g−1), and PC (48 2 0 4 mg of catechin equivalent g−1) compared to Pre-SG, Pre- CC and nontreated samples. Trichoderma harzianum treatment can enhance the hypoglycaemic properties, PC and TPC of Cinnamon extracts and provide new insights into the recovery of phytochemicals.Item Nutritional quality of Cavendish banana (Musa acuminata, AAA) as affected by basil oil and determination of basil oil residues by GC-MS(Journal of Science, Faculty of Science, University of Kelaniya, 2019) Siriwardana, H.; Abeywickrama, K.; Kannangara, S.; Jayawardena, B.The effectiveness of basil oil on the nutritional properties of Cavendish banana and chemical composition of basil oil and oil residue levels of treated banana fruits were evaluated in this study. Cavendish banana hands were treated with 1% alum (w/v), 1% alum (w/v) + 0.4% Ocimum basilicum (basil) oil, distilled water (control) and packaged in Low Density Polyethylene (LDPE) bags and stored at a cold room at 12-14ºC. After two weeks of cold storage banana were induced ripened and nutritional contents of treated Cavendish banana were determined. Gas Chromatography - Mass Spectrometry (GC-MS) was instrumental in identifying the chemical constituents of basil oil as well as residues in basil oil treated Cavendish banana peel after two weeks of storage at 12-14ºC.Item Isolation and identification of litter decomposing fungi in Nagacholai Forest Reserve, Mullaitivu, Sri Lanka(4th International Research Symposium on Pure and Applied Sciences, Faculty of Science, University of Kelaniya, Sri Lanka, 2019) Bopitiya, B. D. S. S.; Dewangani, H. G. N.; Perera, H. W. K.; Premachandra, J. A. H. P.; Senevirathna, M. A. T. M.; Kalpani, N. N.; Kodituwakku, T. D.; Kannangara, S.Nagacholai Forest Reserve, which was disturbed by local terrorism before 2009, is located in Mullaitivu District in the Northern Province of Sri Lanka. It is a tropical dry zone forest, administrated by the Department of Forest Conservation, Sri Lanka. Litter-fall and subsequent nutrient release from decomposing litter are crucial processes that regulate the nutrient availability and the nutrient cycling in soil, as well as the plant growth in forest ecosystems. Litter decomposition rates of forest ecosystems are governed by both biotic and abiotic factors. Therefore, the present study was conducted to study soil characteristics such as pH and electrical conductivity and to isolate and identify the litter decomposing fungi of the Nagacholai forest. Soil and litter were collected randomly by placing 0.5 m X 0.5 m size quadrats on the forest floor. The pH and electrical conductivity of each soil sample were determined by following standard methods. Litter decomposing fungi were isolated from leaf discs (1 mm X 1 mm) obtained from the collected litter following washing and plating method using sterilized distilled water. Pure cultures of isolated fungi were maintained at 28 °C in PDA and morphologically identified using identification keys. The low moisture content in the soil was reflected by brown colour, sandy-podzolic soil. The high proportion of less decomposed, dense litter layer reflected the low decomposition rate due to decelerated soil microbial activity and low water availability. Soil pH was within the range of 6.34 – 6.80 and electrical conductivity at 30 °C varied between 1.23 – 1.57 mS cm-1. Five different fungal species were isolated in high frequencies (50%>) from leaf litter, while four of them were identified up to the genus level based on their vegetative and reproductive morphology (i.e. Mucor sp., Aspergillus sp., Candida sp. and Pestalotiopsis sp.). The remaining unidentified species produced white colour colonies and branched, hyaline, septate hyphae without showing any sexual reproductive structure. Mucor, Aspergillus and Pestalotiopsis are some of the common fungal decomposers found in most of the forests of different climatic zones. However, Candida is a filamentous yeast, which is not frequently reported as a decomposer in terrestrial ecosystems, but found to be a member of the normal soil microflora in forests as secondary sugar fungi. Therefore, Mucor, Aspergillus and Pestalotiopsis could be identified as the most prominent genera of litter decomposing fungi in the Nagacholai forest. Nevertheless, this preliminary study needs to be elaborated further to confirm their identity up to species level using molecular techniquesItem Nutritional quality of Cavendish banana (Musa acuminata, AAA) as affected by basil oil and determination of basil oil residues by GC-MS(Journal of Science of the University of Kelaniya Volume:10, 2019) Siriwardana, H.; Abeywickrama, K.; Kannangara, S.; Jayawardena., B.The effectiveness of basil oil on the nutritional properties of Cavendish banana and chemical composition of basil oil and oil residue levels of treated banana fruits were evaluated in this study. Cavendish banana hands were treated with 1% alum (w/v), 1% alum (w/v) + 0.4% Ocimum basilicum (basil) oil, distilled water (control) and packaged in Low Density Polyethylene (LDPE) bags and stored at a cold room at 12-14ºC. After two weeks of cold storage banana were induced ripened and nutritional contents of treated Cavendish banana were determined. Gas Chromatography - Mass Spectrometry (GC-MS) was instrumental in identifying the chemical constituents of basil oil as well as residues in basil oil treated Cavendish banana peel after two weeks of storage at 12-14ºC.1 Nutritional properties of basil oil treated Cavendish banana showed no adverse changes compared to control. Methyl chavicol (estragole) was the most abundant component (74.44%) of basil oil followed by linalool (15.01%). GC-MS data revealed that negligible amount of residues of basil oil retained in treated Cavendish banana after 14 days. Basil oil treatment and subsequent modified atmosphere packaging of Cavendish banana is recommended as an ecofriendly strategy for air freight or long distance transport over land.Item Nutritional quality of Cavendish banana (Musa acuminata, AAA) as affected by basil oil and determination of basil oil residues by GC-MS.(Journal of Science 2019, Department of Botany, University of Kelaniya, Sri Lanka, 2019) Siriwardana, H.; Abeywickrama, K.; Kannangara, S.; Jayawardena, B.The effectiveness of basil oil on the nutritional properties of Cavendish banana and chemical composition of basil oil and oil residue levels of treated banana fruits were evaluated in this study. Cavendish banana hands were treated with 1% alum (w/v), 1% alum (w/v) + 0.4% Ocimum basilicum (basil) oil, distilled water (control) and packaged in Low Density Polyethylene (LDPE) bags and stored at a cold room at 12-14ºC. After two weeks of cold storage banana were induced ripened and nutritional contents of treated Cavendish banana were determined. Gas Chromatography - Mass Spectrometry (GC-MS) was instrumental in identifying the chemical constituents of basil oil as well as residues in basil oil treated Cavendish banana peel after two weeks of storage at 12-14ºC.1 Nutritional properties of basil oil treated Cavendish banana showed no adverse changes compared to control. Methyl chavicol (estragole) was the most abundant component (74.44%) of basil oil followed by linalool (15.01%). GC-MS data revealed that negligible amount of residues of basil oil retained in treated Cavendish banana after 14 days. Basil oil treatment and subsequent modified atmosphere packaging of Cavendish banana is recommended as an ecofriendly strategy for air freight or long distance transport over land.
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