International Postgraduate Research Conference (IPRC)http://repository.kln.ac.lk/handle/123456789/1552024-03-29T14:16:31Z2024-03-29T14:16:31ZAddition of two Trichoderma species with organic fertilizer paste - A boon for crop yield of Abelmoschus esculentus L. cv. MI 5Kalpani, N. N.Kannangara, B. T. S. D. P.Ratnayake, R. M. C. S.http://repository.kln.ac.lk/handle/123456789/259462023-02-03T08:24:01Z2022-01-01T00:00:00ZAddition of two Trichoderma species with organic fertilizer paste - A boon for crop yield of Abelmoschus esculentus L. cv. MI 5
Kalpani, N. N.; Kannangara, B. T. S. D. P.; Ratnayake, R. M. C. S.
Sustainable agriculture is a farming technique that minimizes environmental impacts while providing a tenable yield. The use of organic amendments as an alternative to inorganic treatments has more significant potential to establish a self-sustaining, less expensive, and environmentally friendly agricultural system. The amalgamation of organic fertilizer with bio-controlling microorganisms is more beneficial than individual application in cropping land to enhance crop productivity. The present study was aimed to examine the influence of an organic fertilizer paste enriched with Trichoderma spp. to enhance the growth performances and yield of Abelmoschus esculentus L. cv. MI 5. The organic fertilizer paste was prepared by aerobic digestion of air-dried and powdered immature twigs of the following plants; Annona glabra, Clidemia hirta, Chromolaena odorata, and Pongamia pinnata (2.0 kg each) in distilled water (42.0 L) for a month. Bio-controlling agents Trichoderma harzianum (KT852821.1) and Trichoderma virens (KP985643.1) were formulated in solid carrier material (compost, straw, clay, and cow urine; 2:1:1:1) separately. The pot trial consisted of six treatments of liquid organic fertilizer enriched with Trichoderma spp. (T10H, T10V, T20H, T20V, T25H, and T25V, where 10, 20, and 25 denote 10%, 20%, and 25% of C. odorata, A. glabra, C. hirta, and P. pinnata extract combined with H as T. harzianum and V as T. virens) with 15 replicates in a completely randomized block design. One-week-old A. esculentus L. cv. MI 5 seedlings were soil treated for 3 months (1st week - 5 mL, 2nd week - 10 mL, 3rd week - 15 mL, 4th week - 20 mL, and 100 mL). The positive and negative controls were commercial fertilizer (Maxicrop) and tap water, respectively. Shoot growth performances, root growth performances, average fresh weights, and average dry weights, and the amount of harvest of A. esculentus L. cv. MI 5 were measured after 3 months of the plantation. One-way ANOVA statistical method, along with Tukey’s multiple comparison tests were used to identify the significant differences (P≤0.05) in growth parameters among treatments using MINITAB (Version 17). T10H treatment (10 % diluted C. odorata, A. glabra, C. hirta, and P. pinnata extract only incorporated with T. harzianum) recorded significantly (P≤0.05), the highest average plant shoot height (163.6±5.40 cm), number of leaves (39±2), stem circumference (5±0.19 cm), average leaf area (309.56±1.2 cm2), root length (38±2.20 cm), the girth of the root (5.24±0.32 cm), number of lateral roots (59±2.08), fresh weight of the entire plant (146.13±16.79 g/plant), fresh weight of the root (35.53±5.82 g/plant), average dry weight of the whole plant (17.61±1.79 g/plant), dry weight of the shoot biomass (13.1±1.42 g/plant), dry weight of the root biomass (4.19±0.09 g/plant), the average number of pods per plant (30±0.24), and average fresh weight of pods (39.83±2.14 g). Therefore, T10H treatment can be successfully used as the best organic fertilizer paste enriched with T. harzianum to enhance the growth and yield of Abelmoschus esculentus L. cv. MI 5.
2022-01-01T00:00:00ZEffect of coconut milk on intestinal barrier function and management of oxidative stressAmbanpola, N.Anjali, N. V. P.Manilgama, T.Gunawardane, M.Seneviratne, K. N.Jayathilaka, N.http://repository.kln.ac.lk/handle/123456789/259452023-02-03T08:22:58Z2022-01-01T00:00:00ZEffect of coconut milk on intestinal barrier function and management of oxidative stress
Ambanpola, N.; Anjali, N. V. P.; Manilgama, T.; Gunawardane, M.; Seneviratne, K. N.; Jayathilaka, N.
Coconut milk (CM) is a major source of dietary fat in a Sri Lankan meal. It is rich in saturated, medium-chain fatty acids (MCFA) and various polyphenols. Some of the ingested fats and polyphenols are not absorbed in the small intestine and reach the colon. This study assessed the formation of metabolic products from CM and the influence of CM on intestinal barrier function. Twelve-week-old female Wistar rats were housed at 25 ± 1°C with a 12 h light and dark cycle. Rats were randomly assigned to two experimental groups (12 rats/ group). Ad libitum access to water and a diet containing 4.2 % total fat; from that 3% fat by means of soybean oil (SOD) control or CM (CMD) was provided for four weeks. Six rats from each group fasted for 10–12 h and were treated with ethanol (20%, 6 g/kg body weight) by oral gavage (SODM and CMDE groups were obtained). Blood (1 mL) was then drawn from the tail vein. Plasma antioxidant capacity, lipid peroxidation, and protein carbonyl content were determined by 2,2-diphenyl-1-picryl-hydrazy (DPPH) assay, ferric reducing antioxidant power (FRAP) assay, protein carbonyl assay, and thiobarbituric acid reactive substances (TBARS) assay according to previously reported methods. At the end of the feeding experiments, animals were subjected to barbiturate euthanasia and a transverse abdominal incision was made. The cecal wash samples with phosphate-buffered saline (pH 7.4) were stored at -80 °C. Liver and brain samples were also harvested. All experimental procedures were approved by the Ethics Review Committee, University of Kelaniya. Short-chain fatty acids (SCFAs) in the cecal wash, plasma, liver, and brain samples were quantified by Gas chromatography. SCFA levels were determined by the standard curves of each SCFA. Shapiro-Wilk normality test (P<0.05) and t-test was used for the statistical comparison. Acetate, propionate, and butyrate concentrations were 802.9±0.4 μg/mL, 156.3±2.1 μg/mL, 20.5±0.4 μg/mL and 802.8±0.4 μg/mL, 153.5±1.7 μg/mL, 19.9±0. μg/mL in CMD and SOD cecal wash samples respectively. Acetate, propionate, and butyrate concentrations were 236.2±0.1 μg/mL, 16.2±0.2 μg/mL, 1.3±0.0 μg/mL and 226.3±1.4 μg/mL, 14.4±0.2 μg/mL, 1.2±0.0 μg/mL in CMD and SOD plasma samples respectively. There was a significant (P<0.05) difference between plasma acetate and propionate levels in CMD compared to SOD. SCFAs were not detected in liver and brain samples. Saccharolytic microbes ferment oligo- and polysaccharides and produce SCFAs. Following their production SCFAs are rapidly absorbed by colonic cells and those not metabolized by colonic cells pass into the liver. Thus, only a small amount of the SCFAs reach systemic circulation and other tissues. Alcohol causes oxidative stress by releasing reactive oxygen species (ROS) during alcohol metabolism. Polyphenols serve as exogenous antioxidants, and they scavenge free radicals to control ROS. According to the four assays, there were no significant differences in the antioxidant capacity between the four groups suggesting no antioxidant effect of coconut milk over soy oil control. Thus, CM has a significant (P<0.05) impact on SCFAs passing through the intestinal barrier but no effect on the management of oxidative stress than soy oil.
2022-01-01T00:00:00ZEffect of the wet extraction methods on the phenolic profile of coconut oilAnjali, N. V. P.Algama, C. H.Seneviratne, K. P.Jayathilaka, N.Seneviratne, K. N.Sakalasuriya, D. D.Silva, C. D.http://repository.kln.ac.lk/handle/123456789/259442023-02-03T08:21:24Z2022-01-01T00:00:00ZEffect of the wet extraction methods on the phenolic profile of coconut oil
Anjali, N. V. P.; Algama, C. H.; Seneviratne, K. P.; Jayathilaka, N.; Seneviratne, K. N.; Sakalasuriya, D. D.; Silva, C. D.
There are multiple methods for producing virgin coconut oil, which can broadly be divided into wet and dry processes. In the wet methods, coconut oil is directly extracted from the coconut milk, an aqueous emulsion is prepared using freshly grated coconut kernel. The method used to extract oil can affect the quality parameters and the phenolic profile of each coconut oil. Therefore, the phenolic profile, and the antioxidant capacity of coconut oil produced using four wet extraction methods, namely, boiling method (BM), fermentation method (FM), chilling and thawing method (CTM) and centrifugation method (CM) were quantified using previously reported methods. The shelf life of each oil sample at 28 ℃ was analyzed based on the induction time of each oil sample using a Rancimat apparatus. Phenolic profiles and unsaponifiable matter were analyzed qualitatively and quantitatively using HPLC. Shelf life at 28 ℃ (2.9±0.0 years), -tocopherol (78.9±0.4 mg/Kg), total phenolic content (660±1 gallic acid equivalent mg/oil Kg) and antioxidant activity (19.4± 1.0%) are significantly (P<0.05) higher in the oil prepared by BM compared to the other wet extraction methods. The phenolic profile of CM and CTM included p-hydroxybenzoic acid, epigallocatechin gallate (EGCG), and epicatechin. The phenolic profile of coconut oil prepared by FM included gallic acid, p-hydroxybenzoic acid, EGCG, epicatechin, and epigallocatechin (EGC). In addition to the p-hydroxybenzoic acid and gallic acid, gallocatechin gallate (GCG), and catechin were found in significantly (P<0.05) higher amounts in coconut oil extracted using BM as a result of epimerization of EGCG and epicatechin to GCG and catechin under the heating conditions used in the BM. Hydrolysis of EGCG was found to be responsible for the observed low levels of EGCG (0.01±0.00 mg/oil Kg) and the presence of gallic acid and EGC in the coconut oil prepared by FM compared to the other two cold extraction methods (CTM, CM). Therefore, the extraction method has a significant impact on the phenolic profile of coconut oil.
2022-01-01T00:00:00ZApplication of rolling circle amplification (RCA) to detect direct amplification of dengue virus in patient serum samples.Manilgama, T.Seneviratne, K. N.Jayathilaka, N.http://repository.kln.ac.lk/handle/123456789/259432023-02-03T08:20:07Z2022-01-01T00:00:00ZApplication of rolling circle amplification (RCA) to detect direct amplification of dengue virus in patient serum samples.
Manilgama, T.; Seneviratne, K. N.; Jayathilaka, N.
Rolling Circle Amplification (RCA) is an isothermal amplification process that can be utilized for rapid amplification of target nucleic acids. In contrast to PCR, which uses thermocycling to mediate denaturation, annealing, and subsequent extension, RCA can be performed at a single reaction temperature making RCA an attractive solution for disease diagnosis based on amplification of pathogen nucleic acids at resource limited settings. In addition, PCR-based detection of pathogenic RNA involves the additional steps required to make complementary DNA copies of the target for amplification. Dengue is a mosquito vector borne viral RNA infection that largely affects urban and semi-urban, sub-tropical and tropical areas. While majority of dengue fever patients recover with careful hospital monitoring some patients may develop severe complications that result in mortality. Therefore, early diagnosis is critical for screening the patients that require hospital management and to prevent exceeding hospital capacity during a dengue outbreak. We developed a direct RCA of dengue virus RNA in serum samples from dengue virus positive patients using Phi 29 DNA Polymerase for the disease diagnosis at resource limited settings. Serum samples were collected from patients suffering from dengue virus infection based on a positive NS 1 antigen test within four days from fever onset with informed consent (n=3). Serum samples collected from healthy individuals were used as the controls (n=3). Multiple Displacement Amplification (MDA) was used to increase the amplification efficiency. Positive control reactions were carried out using a circularized 66 bp linear 5´phosphorylated probe that contained a complementary sequence to all four dengue serotypes and a forward primer against the conserved target region on the probe at 30 oC overnight. The product formation was confirmed by gel electrophoresis following restriction enzyme digestion of the RCA/MDA products with EcoRI. The RCA/MDA products were quantified using a ssDNA dye. Direct isothermal amplification of dengue virus from serum samples collected from dengue infected subjects confirmed that RCA/MDA reaction specifically amplifies dengue virus in patients while no amplification was detected for the serum samples collected from healthy volunteers. Since RCA/MDA can be used for direct gene expression analysis of mRNA and micro RNA in resource limited settings, this novel method can be used for simultaneous disease diagnosis and early prognosis of severe dengue based on differential expression in resource limited settings.
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