ICAPS 2023
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Item Comparison of total chlorophyll, carotene and phenolic contents between species/varieties of Sri Lankan green leafy vegetables Sesbania grandiflora, Centella asiatica and Ipomea aquatica(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Dahanayaka, L. W.; Kadigamuwa, C. C.; Mapa, S. T.Green Leafy Vegetables (GLVs) are rich sources of phytochemicals which are beneficial to reduce chronic non-communicable diseases due to their therapeutic properties such as antioxidant activity. Among numerous dietary supplements of phytochemicals, GLVs are under spotlight as they are rich sources of phenolics and natural pigments including chlorophylls and carotenoids. Availability and the amount of these phytochemicals may vary depending on many factors including the variety of GLVs. The aim of this study is to quantitatively analyze the phytochemical contents; total phenol, chlorophyll and carotene among 3 selected GLV species and 2 varieties of each, Sesbania grandiflora, Centella asiatica and Ipomea aquatica. Each home garden grow variety was acquired from same geographical location which have similar growth conditions. Edible parts of each sample were well washed, air dried at room temperature under shade, and oven dried at 45 ºC to obtain a constant weight. Dried samples were ground and tests were carried out using homogenous sample powder. Total chlorophyll and carotene contents were determined by measuring the absorbance (at the wavelengths of 470 nm, 645 nm and 662 nm) of methanolic extracts and using formulas. Total chlorophyll content in leafy vegetables ranged from 13.75 to 35.56 μg/g dry weight of the sample. The highest chlorophyll content was observed in S. grandiflora variety 1 (V1), whereas the lowest chlorophyll content was observed in I. aquatica variety 2 (V2). The carotene content of studied leaf samples was within the range of 1.08 to 3.89 mg/g dry weight of the sample. S. grandiflora V1 had the highest carotene content whereas the lowest carotene content was observed in C. asiatica V1. Total Phenolic Content (TPC) was determined by the Folin-Ciocalteu method by measuring the absorbance at 760 nm using the methanolic extracts of leaf samples. TPC were within the range of 3.62 to 18.19 mg GAE/g dry weight of the sample. S. grandiflora V1, leaves showed the highest TPC as 18.19 mg GAE/g dry weight of the sample and the lowest TPC was observed in C. asiatica V2 as 3.62 mg GAE/g dry weight of the sample. The results were analyzed in one-way analysis of variance ANOVA test, which showed a statistically significant difference between species/varieties (pItem Preliminary study of antioxidant and anti-inflammatory properties of mistletoe (Dendrophthoe falcata) on mango host plant (Mangifera indica).(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Raseena, M. T. F.; Kadigamuwa, C. C.Dendrophthoe falcata is a common mistletoe plant that has widespread use in Sri Lankan traditional medicines. With the escalating incidence of cancer, asthma, and chronic inflammatory ailments among patients, the exploration of natural remedies in developing nations has become increasingly important. Despite existing research focusing on the chemical composition and biological activities of mistletoe plants in general, a critical gap exists in understanding the specific anti-inflammatory and antioxidant properties of Dendrophthoe falcata when it thrives on its particular host, Mangifera indica. This preliminary study aims to investigate the antioxidant and anti-inflammatory properties of mistletoe, Dendrophthoe falcata leaves grown on the host, Mangifera indica in Sri Lanka. The methanolic extracts of Dendrophthoe falcata and Mangifera indica leaf samples were prepared using the cold extraction method. The antioxidant activity was assessed through the Folin-Ciocalteu assay, ammonium colourimetric method, 2,2- diphenylpicrylhydrazyl (DPPH) assay, and ferric-reducing power assays. Anti-inflammatory action was evaluated using heat-induced hemolysis and albumin denaturation assays. Pearson’s correlation test was performed to determine whether there is a linear relationship between total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity, antiinflammatory properties of Dendrophthoe falcata and Mangifera indica. A two-sample t-test was done to compare the results of TPC and anti-inflammatory and antioxidant properties of Dendrophthoe falcata and Mangifera indica to identify whether these values of mistletoe significantly vary from the host. TPC and TFC of mistletoe were determined to be 1531.0 ± 44.3 mg of gallic acid equivalents per g of extract and 210.7 ± 15.9 mg of catechin equivalents per g of extract through Folin-Ciocalteu assay, ammonium colourimetric method respectively. Mistletoe exhibited an IC50 of 239.70 ± 27.99 µg/mL for DPPH scavenging activity and an EC50 of 6921.0 ± 2419.0 µg/mL for ferric-reducing power. Additionally, mistletoe demonstrated an IC50 of 554.1 ± 42.4 µg/mL for antihemolytic activity and 0.45 ± 0.05 mg/mL for protection against albumin denaturation. Pearson’s correlation analysis revealed that phenols contribute to antioxidant activities. Furthermore, the presence of phenols and flavonoids contributed to the antiinflammatory properties. A t-test comparison between mistletoe and its host plant for bioactive compounds, antioxidants, and anti-inflammatory properties indicated that mistletoe’s bioactive compounds and their antioxidant and anti-inflammatory properties are significantly different from host plants. Dendrophthoe falcata leaves emerge as a promising source of anti-inflammatory and antioxidant properties. This avenue of study bears substantial potential for unlocking novel therapeutic benefits and can be further pursued by determining the chemical structure of the specific bioactive compounds responsible for anti-inflammatory and antioxidant properties.Item Characterization of adsorption properties of naturally occurring clay deposits in Mannar District to remove lead and cadmium from aquatic systems(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Chandrasiri, D. K.; Kadigamuwa, C. C.Water is a fundamental resource for life on earth. Water pollution by heavy metal contamination has a major negative impact on human health due to its toxicity. To remove heavy metals from aquatic environments several methods have been implemented. From these methods, use of natural clays for adsorption of heavy metals is considered to be environmentally friendly and cost effective method. Montmorillonite is one major type of natural clays which is well known for its adsorption ability. Therefore, this study was basically focused on assessing the potential of lead and cadmium adsorption by MMT-rich clay in different places in Sri Lanka. Three soil samples from selected places of Mannar District were collected and conditions were optimised for Pb2+ and Cd2+ adsorption. Then the adsorption was assessed using Atomic Adsorption Spectrophotometer. 100 mg/L initial concentration, 0.10 g adsorbent dosage, and pH 4 as optimum conditions for Pb2+ adsorption, whereas 100 mg/L initial concentration, 0.10 g adsorbent dosage, and pH 6 as optimum conditions for Cd2+ adsorption were observed. Langmuir constants obtained for Pb2+ and Cd2+ were -1.233 L/mg and 0.0477 L/mg. Freundlich constants obtained for Pb2+ and Cd2+ were 2.489 mg/g and 0.764 mg/g respectively. Based on the values obtained for the constants, 1/n, qmax, and RL, it was assumed that both metal ions show multilayer adsorption on a heterogenous surface, which favours Freundlich isotherm. Optimized conditions were applied to the soil samples collected from Giant’s tank, Marvelankeni, and Allavankai areas areas of Northern province. Reduction percentages obtained for adsorption of Cd2+ by soil samples from Giant’s tank, Marvelankeni, and Allavankai areas were 91.385%, 98.381%, 68.152% respectively and adsorption capacities were 111.239 mg/g, 119.755 mg/g, 82.969 mg/g respectively. Reduction percentages obtained for adsorption of Pb2+ by soil samples from Giant’s tank, Marvelankeni, and Allavankai were 48.472%, 30.561%, 58.481% respectively and adsorption capacities were 211.807 mg/g, 133.561 mg/g, 255.286 mg/g respectively. The values obtained were compared with the control sample (reduction percentages 75.633%, 83.751% respectively for Cd2+ and Pb2+), adsorption capacities 92.076 mg/g and 365.986 mg/g respectively for Cd2+ and Pb2+. Based on these results, there might be a potential of using Montmorillonite rich soil in Mannar district to remove Pb2+ and Cd2+ from aquatic systems.Item Preliminary study of anti-inflammatory action of Sri Lankan “dasapanguwa”(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Seneviratne, M. D. I. H.; Kadigamuwa, C. C.The herbal mixture known as “Dasapanguwa” which is frequently used as a traditional medicine in Sri Lanka, contains ten components, including the dried rhizome of Zingiber officinale (Inguru), dried stem of Gerontogea corymbosa (Pathpadagam), dried berries Solanum xanthocarpum (Katuwalbatu), dried seeds of Coriandrum sativum (Koththamalli), dried leaves of Justicia adhatoda (Pawatta), dried stem of Tinospora cordifolia (Rasakinda), dried stem of Coscinium fenestratum (Venivalgata), dried rhizome of Alpinia calcarata (Araththa/Snap ginger), dried stem of Glycyrrhiza brachycarpa (Valmee) and dried stem of Solanum melongena (Eggplant) as most common combination. The objectives of this project were to extract the chemical constituents of each ingredient and the dasapanguwa mixture decoction, to determine the anti-inflammatory action of each extract in dasapanguwa as well as dasapanguwa mixture using the “Heat-Induced Hemolysis” assay. In the present study, the individual ingredients were extracted by deionized water and the dasapanguwa mixture decoction was screened for their anti-inflammatory assay; Heat-Induced Hemolysis. Water extracts were collected according to the ayurvedic traditional method. According to the obtained results of the anti-inflammatory assay, the ability to inhibit protein denaturation (anti-inflammatory activity) varied in the order of dasapanguwa mixture 523.2±31.0 μg/mL > Glycyrrhiza brachycarpa 560.8±59.8 μg/mL > Alpinia calcarata 572.9±18.6 μg/mL > Coscinium fenestratum 787.9±39.4 μg/mL > Solanum xanthocarpum 1634.3±39.0 μg/mL > Gerontogea corymbosa 1880.4±312.2 μg/mL > Tinospora cordifolia 3662.2±366.4 μg/mL > Justicia adhatoda 4184.9±109.6 μg/mL > Zingiber officinale 4290.3±991.9 μg/mL > Coriandrum sativum 5531.7±1883.0 μg/mL > Solanum melongena 6930.3±45.5 μg/mL. The highest IC50 value resulted in the dasapanguwa mixture and the individual extraction of Solanum melongena resulted in the least IC50 value. This study supported the traditional ayurvedic practice of using “Dasapanguwa” as a good source of anti-inflammatory activity. The combination of ten herbals is given higher anti-inflammatory activity than individual herbals.Item Assessment of contaminant levels of hazardous toxic metals and subsequent risk analysis in agricultural soil and selected comestibles within a region afflicted by CKDu in Sri Lanka(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Perera, M. A. S. N.; Kadigamuwa, C. C.; Gunathilaka, V. H. N. M.; Samaraweera, G. A.; Perera, W. P. R. T.; Perera, P. L. R. A.; Liyanage, J. A.; Premaratne, W. A. P. J.Agricultural commodities constitute a pivotal component of the human daily sustenance, yet they are susceptible to contamination by pernicious heavy metals, notably Hg, Cd, Pb, and As. Consequently, agricultural products represent a substantive source of apprehension for human well-being and health. This study aims to evaluate related risk indices of agricultural soils (n=25) and selected comestible grains, namely Oryza sativa (n=25), Vigna radiata (n=25), Vigna unguiculata (n=25), and Vigna unguiculata subsp. (n=25), within the endemic region of Nikawewa Grama Niladhari Division (GND) in North Western Province (NWP) and the reference region, Wewagedara GND in NWP where no evidence for Chronic Kidney Disease of unknown etiology (CKDu). The concentrations of metals in the collected samples were determined by using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and based on that risk indices were calculated. The mean geo-accumulation index (Igeo) for four metals in soil was As - 0.82, Cd - 1.47, Cu - 0.91, and Pb 1.31. That indicates no anthropogenic activities related to As, Cd and Cu while Pb is responsible for moderate pollution in endemic area. Estimated Daily Intake (EDI) values for these heavy metals were calculated with respect to the all grains. EDI means of As and Pb in the endemic and the reference areas for all grain samples were within the Tolerable Daily Intake (TDI) set by United States Environmental Protection Agency (USEPA) (As-0.300 g/kg/day) and World Health Organization (WHO) (Pb-3.600 g/kg/day) while Cd (EDI in endemic area; Oryza sativa-6.385 g/kg/day, Vigna unguiculata-3.386 g/kg/day and Vigna unguiculata subsp.-3.866 g/kg/day) highly exceeded the tolerable weekly intake (TWI) of European Food Safety Authority (EFSA) (Cd-2.5 g/kg/week) in both areas. EDI of Cr in rice samples (1.138 g/kg/day) in endemic area exceeded the TDI set by EFSA (1.0 g/kg/day) and all other values of Cr were below the TDI in both areas. According to the USEPA (intolerable cancer risk (CR) - > 1 x 10-4) cancer risk exist for all these staple grains. Igeo of Pb in the soil, raising concerns about their bioavailability and uptake by the food crops and EDI were indicated the long-term consumption of these foods especially Oryza sativa poses a serious threat to human health. Implementing proper soil management practices, monitoring heavy metal levels in crops, and raising public awareness about safe consumption practices are crucial steps towards mitigating the risk of CKDu.Item Antimicrobial activity of poly-herbal formula “Dasapanguwa” against several Gram positive bacteria and Gram negative bacteria(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Samaraweera, G. A.; Gunathilaka, V. H. N. M.; Perera, M. A. S. N.; Perera, P. D. V. M.; Wageesha, N. D. A.; Kadigamuwa, C. C.The resistance of bacteria against available antibiotics has become a rising problem worldwide. Therefore, the discovery of alternatives using medicinal plants is widely studied. Although medicinal plants have been studied individually, the impact of herbal formulas was studied rarely. “Dasapanguwa” is a poly-herbal formula prepared as a decoction of 10 different plant parts to use in Ayurvedic medicine in Sri Lanka for colds, fever, and infectious diseases. Although the formula is used as a treatment for infectious diseases in Ayurvedic medicine in Sri Lanka, the antimicrobial action of this preparation was not scientifically assessed. The objective of the research was the determination of the antibacterial activity of the Dasapanguwa formula against Gram positive and Gram negative bacteria. Plant parts of Mollugo cerviana (L.) Ser., Solanum virginianum L., Solanum melongena L., Justicia adhatoda L., Rotheca serrata (L.) Steane & Mabb., Glycyrrhiza glabra L., Zingiber Officinale, Coscinium fenestratum (Gaertn.) Colebr., Piper nigrum L., and Coriandrum sativum L. were collected in dried form. Water extracts of decoction 1 (D1) (plant part: 5.0 g) and Decoction 2 (D2) (plants parts: 5.0 g ground into fine powder) were prepared by refluxing in 500.0 ml of water for two hours at 100 °C followed by freeze-drying. The antibacterial susceptibility test was performed by the Kirby-Bauer disc diffusion method against Gram positive bacteria (Staphylococcus aureus, Bacillus subtilis, and Enterococcus faecalis), and Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae). Antibiotic discs were prepared for each concentration of two decoctions (1.0 to 15.0 mg ml-1). Commercially available tetracycline (30 μg) and ceftriaxone (30 μg) were used as positive controls against Gram’s positive and Gram’s negative bacteria respectively according to the CLSI guideline. The diameter of the clear zones was measured. There were no inhibition zones against all the bacteria for tested concentrations of both decoctions compared to positive controls. Previous studies showed that the antibacterial activity of an herbal extract could depend on the plant, solvent, test pathogens, and concentrations. Furthermore, the activity could depend on the plant itself or the formula of poly-herbal. Therefore, it is recommended to further study the antibacterial activity of both decoctions for higher concentrations from 15 mg ml-1 against selected bacteria. Furthermore, the antibacterial activity could be studied for extracts prepared by different solvents including methanol and ethanol.Item Preliminary studies of antioxidant and anti-inflammatory activities in methanol extracts of mistletoe (Pilila) in guava(Faculty of Science, University of Kelaniya Sri Lanka, 2023) Tharakee, D. H.; Kadigamuwa, C. C.Animal welfare receives great attention as it is highly important for enhancing milk production and microbiological quality. Literature reveals that the apprehension of dairy cow welfare practices among farmers is at a low level and limited studies have been carried out on dairy cattle welfare, in Sri Lanka. This study focused on investigating the impact of cattle management and welfare practices on the microbiological quality of raw milk. A total of 87 dairy cows from 50 tie-stall dairy farms in the Naththandiya Area in Sri Lanka were randomly selected. Three categories of measurements were collected; protocol-based measurements, facility-based measurements and animal-based measurements based on a questionnaire. The microbiological quality of raw milk samples was determined using Total Colony Count (TCC) of bacteria and count of Bacillus cereus. Statistical analyses of data were performed using SPSS 23.0 software. The floor quality was assessed in terms of cleanliness, presence of hoof damageable cracks and the grip. It was found that farmers’ attention on quality of flooring remained low. Improper flooring conditions, including presence of dirt (28%), slippage (62%), and hooves being prone to damage (50%), were observed and correlated (P<0.05), with the prevalence of lameness (12.5%) and hock wounds (12.8%). Based on the survey, concrete flooring was the most prominent bedding type (80%) followed by the soil type (20%). The type of flooring significantly influenced the slipperiness of the shed (P<0.001). A considerable percentage of cows (36.0%) showed signs of wounds, and floor slipperiness significantly (P<0.05) affected the prevalence of wounds. Lower attention on the quality of feeders (14%) and water troughs (12 %) was observed. Positive welfare attitude towards cows was observed in 66% of farmers. The average score of farmer attitudes (6.12) and stockperson attitude obtained, significantly (P<0.001) affected on the floor quality. According to the microbiological analysis, 48 % of the samples were found to be contaminated with Bacillus cereus. The Total Colony Count (TCC) of bacteria and the count of Bacillus cereus varied significantly (P<0.05) across different farms. TCC is significantly correlated (P<0.05) with the floor dirtiness of particular farm. However, there was no significant difference (P>0.05) in between the floor type, TCC and Count of Bacillus cereus. The current study highlighted the critical welfare issues of dairy cows in Naththandiya area and some of these welfare issues had impact on the quality of raw milk. Therefore, it is essential to improve management practices and increase stockperson awareness to enhance dairy cow welfare and there by the quality of raw milk.