Browsing by Author "Manage, P.M."

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    Diversity and Relative Abundance of Industrially Important Enzyme Producing Microbial Genera in Hot Springs in Sri Lanka
    (Uva Wellassa University of Sri Lanka, 2021) Sadeepa, H.D.D.; Sirisena, K.A.; Manage, P.M.
    Thermal tolerant enzyme producing bacteria can be successfully used in industrial settings to minimize environmental impacts caused by excess use of chemical catalysts in production chains. Hot springs are a major habitat for extremophiles which produce extremozymes. Characterization and identification of microbial community in hot springs provide an initial platform for identification of extremophilic microorganisms for industrial applications. In the present study, the diversity and relative abundance of industrially important enzyme producing microbial genera in hot springs in Sri Lanka was studied. Water samples were collected from surface and bottom of the hot springs. Temperature, conductivity, pH and Dissolved Oxygen (DO) were measured at the site using portable meters. To analyse microbial community (bacteria and archaea) of hot springs, extracted DNA was sequenced through 16s rDNA amplicon sequencing on Illumina MiSeq platform. Sequencing data were analyzed using Mother V. 1.42 software. METAGENassist web server tool was used to predict the metabolic functional diversity of the bacterial and archaeal communities. The temperature of the hot springs were ranged from 33.7 0C to 55.4 0C where conductivity, pH and DO levels were reneged from 801 to 1507 μS/cm, 7.20 to 8.27 and 1.05 – 3.5 mg/L respectively. The microbial communities of the hot springs were mostly comprised of Bacteria and Archaea. Bacteria was the dominant component respectively 99% and only <1% Archaea. Chloflexus, Rubelllimicrobium, Acinetobacter, Pseudomonas, Methylobacterium, Tepidimonas, Rheinheimera, Flavobacterium, and Vogesella were the major bacteria genera recorded in all hot springs. The metabolic inference analysis of the microbial community of hot springs comprised of dinitrogen-fixing bacteria, lignin degraders, nitrogen and sulfate reducers, nitrogen fixation bacteria, sulfur reducers, denitrifying bacteria, cellulose degraders and sugar fermentors. Thus the results of the present study implied that the hot springs could be a useful source of bacteria for future industrial perspectives. Further studies are in progress along with a metagenomic analysis. Keywords: Hot springs; Community analysis; Extremophiles; NGS analysis; Biotechnological Prospects
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    In-vitro Study on Geosmin and 2-Methylisoborneol Producing Four Species of Native Cyanobacteria
    (Uva Wellassa University of Sri Lanka, 2019) Ganegoda, S.S.; Wijesekara, W.M.A.; Chinthaka, S.D.M.; Wanniarachchi, D.D.C.; Manage, P.M.
    The production of odiferous metabolites, such as Geosmin and 2-methlyisoborneol (2- MIB) cause muddy and moldy taste and odor in water is a major limitation in safe and adequate provision of potable waters in worldwide. The biological functions of these odorants have not known, and cyanobacteria are the major producers of Geosmin and 2- MIB in natural waters as a result of their secondary metabolites. Objective of the current study was to investigate four laboratory cultured cyanobacteria’s ability to produce Geosmin and 2- MIB. Production of Geosmin and 2- MIB was studied using native cyanobacteria isolated from Unnichchi tank in Baticolloa district, Jayanthi tank in Ampara district and Padawiya tank in Anuradhapura district where taste and odour issues are prevailing. Isolation of cyanobacteria was carried out using dilution and spread plate method by maintaining cultures in BG 11 medium with antibiotics. The cultures were identified as Oscillatoria sp., Psudoanabena sp., Lyngbya sp., and Nostoc sp. using the 16s rRNA sequencing. 10 ml aliquots of two months old axenic cultures were subjected to Solid Phase Micro Extraction (SPME) and Gas Chromatography Mass Spectrometry (GC/MS) analysis with BG 11 culture medium as control. The concentration of Geosmin produced from the cyanobacteria were ranged between 10.8 ng/L to 162.6 ng/L whereas 2- MIB levels ranged from 50.8 ng/L to 532.5 ng/L. Axenic cultures of Nostoc sp. recorded the highest Geosmin level (162.6 ng/L) and highest 2- MIB level was detected from Psudoanabena sp. (532.5 ng/L). Further Nostoc sp. recorded a significant high level of 2- MIB level (379.6 ng/L) compared to Oscillatoria sp. and Lynbya sp. (p < 0.05). It can be concluded that above species are commonly available cyanobacteria species in Sri Lankan water bodies and popular Geosmin and 2- MIB producers.
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    Monitoring Tributyltin (TBT) Contamination of Southern Coastal Waters in Sri Lanka
    (Uva Wellassa University of Sri Lanka, 2019-02) Bandara, K.R.V.; Chinthaka, S.D.M.; Manage, P.M.
    Tributyltin (TBT) is an organotin compound belongs to the group of Persistent Organic Pollutants (POPs) and one of the active ingredient in antifouling paints used for boat hulls, docks, fish nets in order to prevent the growth of aquatic fouling organisms. The presence of TBT in the aquatic environment is extremely toxic to target and non-target organisms and it is responsible for severe sexual disorders like sex changes called imposex leading to sterile aquatic populations which lead to decline of animal diversity. According to WHO, the No Observed Effective Level (NOEL) of TBT is below 1 ngL-1. However, there are no information has been recorded regarding the TBT contamination in water and its adverse effect on aquatic organisms in Sri Lanka. Therefore, the study aims to monitor the contamination status of TBT in southern coastal area in Sri Lanka. Coastal water samples were collected from river canals, fishing harbors and river mouth along the Southern Coastal area from Dehiwala to Mirissa. TBT was extracted by using a newly optimized Solid Phase Micro Extraction (SPME) method. Hydridization process was followed for quantification by using the Gas Chromatography-Mass Spectrometry (GCMS) in parts per trillion level (ppt). Results showed that the highest TBT concentration in highly polluted Dehiwala (3.4ngL-1) and Wellawatta canals (3.4ngL-1). TBT concentrations in fishing harbors at Ambalangoda, Galle, Mirissa and Beruwala were range from 1.4 ngL-1 to 3.2ngL-1 where in Madu and Benthota river mouth were 0.8ngL- 1 and 0.9ngL-1 respectively. The recovery of the TBT extraction method was 87±0.1% for the artificial sea water. Thus, the preliminary results of the study showed that high contamination of TBT may adversely effect on marine biodiversity.
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    Nephrotoxic Effect of Environmental Water Sample on Wistar Rats with Special Reference to Synergistic Effect of Fluoride and Hardness
    (Uva Wellassa University of Sri Lanka, 2021) Dilrukshi, K.T.; Wanigasuriya, K.; Beneragama, D.H.; Suresh, T.S.; Manage, P.M.
    Chronic Kidney Disease of unknown aetiology (CKDu) is a serious health problem in Sri Lanka and it has spread in epidemic proportions among the rural farming communities in the North Central Dry Zone of the country. There are several hypotheses on this phenomenon and the present study was focused on the nephrotoxic effect of environmental water samples on Wistar rats. Special attention was taken to evaluate the synergistic effect of water hardness and fluoride on nephrotoxicity. Experimental protocols were approved by the ethics committee of the Institute of Biology, Sri Lanka (ERCIOBSL 194062019). Fourteen Wistar rats were randomly assigned into two groups (n=7). One group of rats was treated with water sample collected from Padaviya while the other group was given de-ionized (DI) water as the control group. Body weight and daily water consumption of each rat were measured and blood and urine samples were collected at 0, 7, 14, 28, 42, 60, 90, 120 and 150 days‟ intervals. Serum creatinine, urine creatinine and urinary biomarker KIM-01 were analysed. The concentrations of fluoride and hardness of Padaviya water sample were 1.91 and 280 mg/L respectively. There was no significant difference in body weights and daily water consumption between treated and control groups (p>0.05) were recorded. Average serum creatinine levels of the rats treated with Padaviya water and DI water were ranged from 0.73 to 1.02 mg/dL and 0.56 to 0.71 mg/dL respectively. Average urine creatinine levels of the rats treated with Padaviya water and DI water were ranged from 0.61 to 0.76 mg/dL and 0.65 to 0.7 mg/dL respectively. KIM-1 level was not detected in the control group treated with DI water where rats received Padaviya water, the KIM-1 level was 5.9 ng/mL at the end of the study. The results indicate that there was a nephrotoxic effect from Padaviya water sample which recorded high fluoride and water hardness. Thus, further studies are needed to evaluate the synergistic effect of water hardness and fluoride to determine the threshold levels. Keywords: Synergistic effect; Fluoride; Water hardness; Creatinine levels; Nephrotoxic effects
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    Phytochemical Screening, Quantitative Analysis and In Vitro Cytotoxic Potential of Justicia adhathoda Leaf Extracts
    (Uva Wellassa University of Sri Lanka, 2021) Silva, A.B.W.R.; Nugara, N.N.R.N.; Manage, P.M.; Undugoda, L.J.S.; Udayanga, D.; Nilmini, A.H.L.R.
    Justicia adhathoda L., also known as “Adhathoda” is one of the widely used medicinal plants in indigenous medicinal systems in Sri Lanka and India. Previous studies have found several beneficial effects of different solvent extracts of this plant. However, the best solvent to extract much of the phytoconstituents has not been studied. Therefore, the present study was focused on identification, quantification of phytoconstituents in different crude extracts and in vitro cytotoxic effect of J. adhathoda. Plant leaves were collected from Ambilipitiya, Sri Lanka. Ethanol (EE), hexane (HE) and water (WE) extracts were prepared by adding 1 g of powdered leaf samples to 25 mL of ethanol (99.9%), n-hexane and distilled water, separately in triplicates. Preliminary phytochemical screening was performed according to the Harborne method. Total phenolic content (TPC) was measured spectrophotometrically by Folin-Ciocalteu assay and total flavonoid content (TFC) was determined using aluminum chloride colorimetric assay. To identify the toxicity level of J. adhathoda, different concentration of the WE were exposed to 3T3-L1 cells, a well-established cell culture model that is extensively used to study adipogenesis. The treatment was carried out for 8 days and performed MTS assay to determine the suitability of the WE consumption. Phytochemical screening results showed the availability of flavonoids, terpenoids, steroids, anthocyanins, alkaloids and coumarins in the WE. In contrast, tannin and alkaloids were abundant in the EE. TPC in the WE was 8-fold higher than the EE. The TFC in the EE was 20-fold higher than HE, and marked the highest. Collectively, the best solvent to extract the maximum amount of phytoconstituents was found as distilled water. The WE showed no toxicity below 500 μg/mL against 3T3-L1 adipocytes. The presence of high content of different phytoconstituents in WE of J. adhathoda is a great advantage to gain the health benefits as herbal tea or herbal porridges. These results provide referential information to identify the medicinal value of J. adhathoda and to be utilized in screening novel target compounds to treat various diseases. Keywords: Justicia adhathoda; phytochemical analysis; Medicinal plant; TFC; TPC; 3T3-L1
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    Screening and Quantification of Tetracycline and Sulfonamide Resistance Genes in Natural Environmental Samples
    (Uva Wellassa University of Sri Lanka, 2019) Liyanage, G.Y.; Manage, P.M.
    Screening of antibiotic resistance is important factor for assessing risk of antibiotic resistance in aquaculture environment. In this study, tetracycline (tet A and tet M) and sulfanomide (sul 1 and sul 2) resistance genes were screened in 24 sampling locations including marine water samples at different levels (100, 500 and 1000 m), lagoons (Chilow, Puttalam), rivers (Kelani, Kalu, Walawe, Mahaweli) and from Horton plains. The Tetracycline (TET and OTC) and Sulfanomide (SUF) contamination levels in the sampling locations were analyzed by using High Performance Liquid Chromatography (HPLC). DNA was extracted and Polymerase Chain Reaction (PCR) method was followed to screen the presence of Antibitic Resistance Genes (ARGs) in the samples. Real time PCR assays were employed to quantify ARGs. None of the samples were shown the contaminations of tetracycline and sulfonamides. The tet (M) gene was detected in 7 sampling sites. From the tested ARGs, both tet A and tet M were found at 3 marine samples (100m), 2 lagoon samples, whereas both sul 1 and sul 2 were recorded at only in one lagoon sample. No resistance genes were found in >100 m depth marine samples and in any river samples. The average concentrations of the Sul 1 and Sul 2 was ranged between 0.12 × 10 – 1.56 × 10 copies/ml, 0.05 × 10 – 0.56 × 10 copies/ml in lagoon samples while tet M and tet A were ranged 0.11 × 10 - 1.23 × 102 copies/ml, 0.13 × 10 – 4.56 × 10 copies /ml in marine samples. The presence of tetracycline and sulphonamide resistance genes may limit the effectiveness of antibiotics in treating fish illnesses, thereby causing a potential risk to the aquaculture industry.