Isolation and Identification of Cellulase Producing and Sugar Fermenting Bacteria for Second-Generation Bioethanol Production

 

biofuel

Authors: L.I. Weerasinghe, Tharindu Madusanka and Pathmalal M. Manage

Journal: International Journal of Renewable Energy Development

Over the last decades, the negative impacts of fossil fuel on the environment and increasing demand for energy due to the unavoidable depletion of fossil fuels, has transformed the world’s interests towards alternative fuels. In particular, bioethanol production from cellulosic biomass for the transportation sector has been incrementing since the last decade. The bacterial pathway for bioethanol production is a relatively novel concept and the present study focused on the isolation of potential “cellulase-producing” bacteria from cow dung, compost soil, and termite gut and isolating sugar fermenting bacteria from palm wine. To select potential candidates for cellulase enzyme production, primary and secondary assays were conducted using the Gram’s iodine stain in Carboxy Methyl Cellulose (CMC) medium and the Dinitrosalicylic acid (DNS) assays, respectively. Durham tube assay and Solid-Phase Micro-Extraction (SPME) coupled with Gas Chromatography-Mass Spectrometry (GC-MS) was used to evaluate the sugar fermenting efficiency of the isolated bacteria. Out of 48 bacterial isolates, 27 showed cellulase activity where Nocardiopsis sp. (S-6) demonstrated the highest extracellular crude enzyme activity of endoglucanase (1.56±0.021 U) and total cellulase activity (0.93±0.012 U). The second-highest extracellular crude enzyme activity of endoglucanase (0.21±0.021 U) and total cellulase activity (0.35±0.021 U) was recorded by Bacillus sp. (T-4). Out of a total of 8 bacterial isolates, Achromobacter sp. (PW-7) was positive for sugar fermentation resulting in 3.07% of ethanol in broth medium at 48 h incubation. The results of the study revealed that Nocardiopsis sp. (S-6) had the highest cellulase enzyme activity. However, the highest ethanol percentage was achieved with by having both Bacillus sp. (T-4) and Achromobacter sp. (PW-7) for the simultaneous saccharification and fermentation (SSF) method, as compared to separate hydrolysis and fermentation (SHF) methodologies

Spatiotemporal distribution of cutaneous leishmaniasis in Sri Lanka and future case burden estimates

journal-pntd-0009346-g001Authors: Nadira D. Karunaweera, Sanath Senanayake, Samitha Ginige, Hermali Silva, Nuwani Manamperi, Nilakshi Samaranayake, Rajika Dewasurendra, Panduka Karunanayake, Deepa Gamage, Nissanka de Silva, Upul Senarath, Guofa Zhou

Journal: PLoS Negl Trop Dis

Leishmaniasis is on the rise in Sri Lanka in contrast to the declining trend in rest of South Asia. Spatiotemporal analysis and disease risk factors are useful for understanding transmission mechanisms and predicting future disease distribution to facilitate control. In this study we analyzed data on cutaneous leishmaniasis cases from Sri Lanka from 2001 to 2019. We asked three important questions regarding the driving forces behind the intensified leishmaniasis transmission: 1) Are the transmission dynamics in different areas synchronized? 2) What is the role of neighboring-area dispersal in shaping transmission dynamics? 3) How important is climatic variability in transmission dynamics? We used a multi-step approach to answer these questions. In addition to cross-correlation analysis, we built a mixed spatiotemporal regression-autoregression model to analyze risk factors, which is unique in leishmaniasis research because the simplified model was also useful for predicting future disease distribution. We found that the incidence dynamics in different districts could be divided into three synchronized groups based on similarity. Risk factor analysis indicated that precipitation, neighboring-district dispersal, and local infection carryover played important roles in shaping transmission dynamics. The spatiotemporal model predicted intensifying transmission with increasing case numbers, and expansion of high-transmission areas. Targeted interventions are urgently needed to stem the outbreak.

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Compensatory Base Changes Reveal Sexual Incompatibility among Members of the Anopheles subpictus Sensu Lato (Diptera: Culicidae) Species Complex in Sri Lanka

life-11-00211-g004-550

Authors: D. P. W. Jayatunga,I. N. Harischandra,N. V. Chandrasekhara and B. G. D. N. K. de Silva

Journal: Life

The mosquito Anopheles (Cellia) subpictus sensu lato (s.l.) is a major secondary vector of malaria in Sri Lanka. The sibling species composition in this species complex in Sri Lanka remains debatable. Compensatory base changes (CBCs) in the secondary structures of internal transcribed spacer 2 (ITS2) are reliable sources to predict sexual incompatibility among closely related species. The objective of the present study was to investigate the An. subpictus s.l. populations in Sri Lanka using the CBC analysis. Mosquito DNA was amplified and sequenced for the ITS2 region. The sequences were annotated using ITS2 Database. ITS2 secondary structures were constructed and analyzed for CBCs using various bioinformatics tools. The ITS2 regions consisted of two different lengths, 575 bp and 480 bp. The two CBCs and three hemi CBCs identified in the present study suggest that there may be at least two sexually incompatible sibling species. In conclusion, it is likely that there may be only two reproductively isolated sibling species in the An. subpictus species complex in Sri Lanka. However, due to high divergence of ITS2 in these species, it is reasonable to assume that they may be undergoing a speciation event to separate as a distinct species.

solation and Molecular Characterization of Polycyclic Aromatic Hydrocarbon Degrading Bacteria from Effluent Water from Weras River Park, Sri Lanka

Authors: A.A. Prathiba, F. Sumaiya Idroos and Pathmalal M. Manage

Journal: Environment and Natural Resources Journal

The present study records the detection of PAHs such as naphthalene and anthracene and isolation of PAHs degrading bacteria from a restaurant site, Weras River Park, in Boralesgamuwa, Sri Lanka. Water samples were collected in seven locations of the study area. Water temperature (oC), pH, and electric conductivity (EC) were measured at the site itself using standard meters. Nitrogen nitrate (N-NO3-) and total phosphate (TP) were measured at the laboratory following the standard methods. Following the extraction of PAHs in collected water samples, detection was carried out using the PDA-HPLC diode array method. PAH degrading bacteria were identified using microplate assay. The selected bacteria strains were subjected to degradation kinetic studies following molecular identification by 16S rRNA analysis. Phylogenetic analysis identified Achromobacter spanius as potential naphthalene degrading bacteria, where Alcaligens faecalis was recorded as an anthracene degrader. Degradation study confirmed that A. spanius efficiently degraded naphthalene at the rate of 0.145±0.002 ppm/day, whereas A. faecalis degraded anthracene at the rate of 0.181±0.036 ppm/day, respectively. Degradation of structures of the Naphthalene and Anthracene by A. spanius and A. faecalis was further analyzed by Fourier Transform Infrared Spectroscopy (FTIR). This is the first record on naphthalene degradation by the bacterium A. spanius.

prathiba