Search results for: eucalyptus clones

Authors: Kien Xuan Ngo

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Cassiicolin (Cas), a toxin produced by Corynespora cassiicola, is responsible for corynespora leaf fall (CLF) disease in rubber trees. Currently, the molecular mechanism of the cytotoxicity of Cas isoforms (i.e., Cas1, Cas2) on rubber leaves and its host selectivity have not been fully elucidated. This study analyzed the binding of Cas1 and Cas2 to membranes consisting of different plant lipids and their membrane-disruption activities. Using high-speed atomic force microscopy and confocal microscopy, this study reveals that the binding and disruption activities of Cas1 and Cas2 on lipid membranes are strongly dependent on the specific plant lipids. The negative phospholipids, glycerolipids, and sterols are more susceptible to membrane damage caused by Cas1 and Cas2 than neutral phospholipids and betaine lipids. In summary, This study unveils that (i) Cas1 and Cas2 directly damage and cause necrosis in the leaves of specific rubber clones; (ii) Cas1 and Cas2 can form biofilm-like structures on specific lipid membranes (negative phospholipids, glycerolipids, and sterols). The biofilm-like formation of Cas toxin plays an important role in selective disruption on lipid membranes; (iii) Vulnerability of the specific cytoplasmic membranes to the selective Cas toxin is the most remarkable feature of cytotoxicity of Cas toxin on plant cells. Finally, researcher’s exploration is crucial to understand the basic molecular mechanism underlying the host-selective toxic interaction of Cas toxin with cytoplasmic membranes in plant cells.

Keywords: cassiicolin, corynespora leaf fall disease, high-speed AFM, giant liposome vesicles

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Authors: M. F. Nawaz, S. Gul, T. H. Farooq, M. T. Siddiqui, M. Asif, I. Ahmad, N. K. Niazi

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In Pakistan, major demands of fuel wood and timber wood are fulfilled by agroforestry. However, the information regarding economic significance of agroforestry and its productivity in Pakistan is still insufficient and unreliable. Survey of field conditions to examine the agroforestry status at local level helps us to know the future trends and to formulate the policies for sustainable wood supply. The objectives of this research were to examine the actual status and potential of agroforestry and to point out the barriers that are faced by farmers in the adoption of agroforestry. Research was carried out in Chiniot district, Pakistan because it is the famous city for furniture industry that is largely dependent on farm trees. A detailed survey of district Chiniot was carried out from 150 randomly selected farmer respondents using multi-objective oriented and pre-tested questionnaire. It was found that linear tree planting method was more adopted (45%) as compared to linear + interplanting (42%) and/or compact planting (12.6%). Chi-square values at P-value <0.5 showed that age (11.35) and education (17.09) were two more important factors in the quick adoption of agroforestry as compared to land holdings (P-value of 0.7). The major reason of agroforestry adoption was to obtain income, fodder and fuelwood. The most dominant species in farmlands was shisham (Dalbergia sissoo) but since last five years, mostly farmers were growing Sufeida (Eucalyptus camaldulensis), kikar (Acacia nilotica) and popular (Populus deltoides) on their fields due to “Shisham die-back” problem. It was found that agro-forestry can be increased by providing good quality planting material to farmers and improving wood markets.

Keywords: agroforestry, trees, services, agriculture, farmers

Procedia PDF Downloads 427

Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

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Authors: Z. Nourmohammadi, F. Farahani, M. Shaker

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Aloe is an important commercial crop available in a wide range of species and varieties in international markets. The applications of this plant have been recorded in the ancient cultures of India, Egypt, Greece, Rome and China. Aloe has been used for centuries and is currently being actively studied for medicinal purposes. Aloe is propagated through lateral buds, which is slow, very expensive and low income practice. Nowadays, it has been cultured by in vitro propagation for rapid multiplication of plants, genetic improvement of crops, obtaining disease-free clones and for progressive valuable germplasm. The present study focused on the influence of different phytohormones on rapid in vitro propagation of Aloe plants. We also investigated the effect of gamma radiation on biochemical characters as well as genetic changes. Shoot tip of 2-3 cm were collected from offshoot of Aloe barbadensis and Aloe littoralis, and were inoculated with MS medium containing various concentrations of BA (0.5, 1, 2 mg/l), IAA (0.5, 1 mg/l). The best treatment for a highest shoot number and bud proliferation was MS medium containing 2 mg/l BAP and 0.5 mg/l IAA in A. barbadensis and A. littoralis. Maximum percentage of proliferated shoot buds (90% and 95%) from a single explant were obtained in MS medium after 4-5 weeks of the second and the first subcultures, respectively. Different genome sizes were also indicated among treatments and subcultures. The mixoploids identified in flow cytometery histograms in different treatments. The effect of gamma radiation on A. littoralis showed that by increasing the dose of gamma radiation, amounts of chlorophyll A, B, carotenoids, total protein content and superoxide dismutase were significantly increased compared to control plants. Genetic variation analysis also revealed significant genetic differences between control and gamma radiation treated regenerated plants by AMOVA test. Higher genetic heterozygocity was observed in radiation treated plants. Our findings may provide useful method for improving of Aloe plant proliferation with increasing of useful material such as antioxidant enzymes.

Keywords: aloe, antioxidant enzyme, micropropagation, gamma radiation, genetic variation

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Authors: Hassan Mahmoudzadeh

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The bacterium of Agrobacterium vitis causes crown and root gall disease, an important disease of grapevine, Vitis vinifera L. Also, Phylloxera is one of the most important pests in viticulture. Grapevine rootstocks were developed to provide increased resistance to soil-borne pests and diseases, but rootstock effects on some traits remain unclear. The interaction between rootstock, scion and environment can induce different responses to the grapevine physiology. 'Sultsna' (Vitis vinifera L.) is one of the most valuable raisin grape cultivars in Iran. Thus, the aim of this study was to determine the rootstock effect on the growth characteristics and yield components and quality of 'Sultana' grapevine grown in the Urmia viticulture region. The experimental design was completely randomized blocks, with four treatments, four replicates and 10 vines per plot. The results show that all variables evaluated were significantly affected by the rootstock. The Sultana/110R and Sultana/Nazmieh were among other combinations influenced by the year and had a higher significant yield/vine (13.25 and 12.14, respectively). Indeed, they were higher than that of Sultana/5BB (10.56 kg/vine) and Sultana/Spota (10.25 kg/vine). The number of clusters per burst bud and per vine and the weight of clusters were affected by the rootstock as well. Pruning weight/vine, yield/pruning weight, leaf area/vine and leaf area index are variables related to the physiology of grapevine, which was also affected by the rootstocks. In general, rootstocks had adapted well to the environment where the experiment was carried out, giving vigor and high yield to Sultana grapevine, which means that they may be used by grape growers in this region. In sum, the study found the best rootstocks for 'Sultana' to be Nazmieh and 110R in terms of root and shoot growth. However, the choice of the right rootstock depends on various aspects, such as those related to soil characteristics, climate conditions, grape varieties, and even clones, and production purposes.

Keywords: grafting, vineyards, grapevine, succeptability

Procedia PDF Downloads 76

Authors: Karem H. Alzoubi, Ahmad S. Alkofahi, Omar F. Khabour, Nizar M. Mhaidat

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Herbal medicinal products represent a major focus for drug development and industry and it holds a significant share in drug-market all over the globe. In here, selected medicinal plant extracts from Jordan with high antioxidative capacity were tested for their protective effect against oxidative DNA damage using in vitro 8-hydroxydeoxyguanisine and sister chromatid exchanges (SCEs) assays in cultured human lymphocytes. The following plant extracts were tested Cupressus sempervirens L., Psidium guajava (L.) Gaerth., Silybum marianum L., Malva sylvestris L., Varthemia iphionoides Boiss., Eminium spiculatum L. Blume, Pistachia palaestina Boiss., Artemisia herba-alba Asso, Ficus carica L., Morus alba Linn , Cucumis sativus L., Eucalyptus camaldulensis Dehnh., Salvia triloba L., Zizyphus spina-christi L. Desf., and Laurus nobilis L. A fractionation scheme for the active plant extracts of the above was followed. Plants extract fractions with best protective properties against DNA damage included hexane fraction of S. marianum L. (aerial parts), chloroform fractions of P. palaestina Boiss. (Fruits), ethanolic fractions of E. camaldulensis Dehnh (leaves), S. triloba L. (leaves), and ethanolic fractions of Z. spina-christi L. Desf. (Fruits/leaves). On the other hand, the ethanolic extracts of V. iphionoides Boiss was found to increase oxidative DNA damage. Results of the SCEs are undergoing. In conclusion, plant extracts with antioxidative DNA damage properties might have clinical applications in cancer prevention.

Keywords: medicinal plants extract, fractionation, oxidative DNA damage, 8-hydroxydeoxyguanisine, SCEs, Jordan

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Authors: Nassima Elyebdri, Asma Boumediou, Soumia Addoun

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Traditional medicine has been part of the Algerian culture for decades. In particular, the city of Tlemcen still retains practices based on phytotherapy to the present day, as this kind of medicine fulfills the needs of its followers among the local population. The toxic plants contain diverse natural substances which supplied a lot of medicine in the pharmaceutical industry. In order to explore new medicinal sources among toxic plants, an ethnobotanical study was carried out on the use of these plants by the population, at Emir Abdelkader Square of the city of Tlemcen, a rather busy place with a high number of traditional health practitioners and herbalists. This is a descriptive and transversal study aimed at estimating the frequency of using toxic plants among the studied population, for a period of 4 months. The information was collected, using self-anonymous questionnaires, and analyzed by the IBM SPSS Statistics software used for statistical analysis. A sample of 200 people, including 120 women and 80 men, were interviewed. The mean age was 41 ± 16 years. Among those questioned, 83.5% used plants; 8% of them used toxic plants and 35% used plants that can be toxic under certain conditions. Some improvements were observed in 88% of the cases where toxic plants were used. 80 medicinal plants, belonging to 36 botanical families, were listed, identified and classified. The most frequent indications for these plants were for respiratory diseases in 64.7% of cases, and for digestive disorders in 51.5% of cases. 11% of these plants are toxic, 26% could be toxic under certain conditions. Among toxics plants, the most common ones are Berberis vulgaris with 5.4%, indicated in the treatment of uterine fibroids and thyroid, Rhamnus alaternus with 4.8% for hepatic jaundice, Nerium oleander with 3% for hemorrhoids, Ruta chalepensis with 1.2%, indicated for digestive disorders and dysmenorrhea, and Viscum album with 1.2%, indicated for respiratory diseases. The most common plants that could be toxic are Mentha pulegium (15.6%), Eucalyptus globulus (11.4%), and Pimpinella anisum (10.2%). This study revealed interesting results on the use of toxic plants, which are likely to serve as a basis for further ethno-pharmacological investigations in order to get new drug sources.

Keywords: ethnobotany, phytotherapy, Tlemcen, toxic plants

Procedia PDF Downloads 290

Authors: Eric York, James Tadio, Silas Owusu Antwi

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Laundry wastewater contains a diverse range of chemical pollutants that can have detrimental effects on human health and the environment. In this study, simulation studies by Spyder Python software v 3.2 to assess the efficacy of biochar in removing PO₄³⁻ from wastewater were conducted. Through modeling and simulation, the mechanisms involved in the adsorption process of phosphate by biochar were studied by altering variables which is specific to the phosphate from common laundry phosphate detergents, such as the aqueous solubility, initial concentration, and temperature using the Dudinin Approach (DA). Results showed that the concentration equilibrate at near the highest concentrations for Sugar beet-120 mgL⁻¹, Tailing-85 mgL⁻¹, CaO- rich-50 mgL⁻¹, Eggshell and rice straw-48 mgL⁻¹, Undaria Pinnatifida Roots-190 mgL⁻¹, Ca-Alginate Granular Beads -240 mgL⁻¹, Laminaria Japonica Powder -900 mgL⁻¹, Pinesaw dust-57 mgL⁻¹, Ricehull-190 mgL⁻¹, sesame straw- 470 mgL⁻¹, Sugar Bagasse-380 mgL⁻¹, Miscanthus Giganteus-240 mgL⁻¹, Wood Bc-130 mgL⁻¹, Pine-25 mgL⁻¹, Sawdust-6.8 mgL⁻¹, Sewage Sludge-, Rice husk-12 mgL⁻¹, Corncob-117 mgL⁻¹, Maize straw- 1800 mgL⁻¹ while Peanut -Eucalyptus polybractea-, Crawfish equilibrated at near concentration. CO₂ activated Thalia, sewage sludge biochar, Broussonetia Papyrifera Leaves equilibrated just at the lower concentration. Only Soyer bean Stover exhibited a sharp rise and fall peak in mid-concentration at 2 mgL⁻¹ volume. The modelling results were consistent with experimental findings from the literature, ensuring the accuracy, repeatability, and reliability of the simulation study. The simulation study provided insights into adsorption for PO₄³⁻ from wastewater by biochar using concentration per volume that can be adsorbed ideally under the given conditions. Studies showed that applying the principle experimentally in real wastewater with all its complexity is warranted and not far-fetched.

Keywords: simulation studies, phosphate removal, biochar, adsorption, wastewater treatment

Procedia PDF Downloads 67

Authors: Liliane Samara Ferreira Leite, Francys Kley Vieira Moreira, Luiz Henrique Capparelli Mattoso

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The increasing environmental concern regarding the plastic pollution worldwide has stimulated the development of low-cost biodegradable materials. Proteins are renewable feedstocks that could be used to produce biodegradable plastics. Gelatin, for example, is a cheap film-forming protein extracted from animal skin and connective tissues of Brazilian Livestock residues; thus it has a good potential in low-cost biodegradable plastic production. However, gelatin plastics are limited in terms of mechanical and barrier properties. Cellulose nanocrystals (CNC) are efficient nanofillers that have been used to extend physical properties of polymers. This work was aimed at evaluating the reinforcing efficiency of CNC on gelatin films. Specifically, we have employed the continuous casting as the processing method for obtaining the gelatin/CNC bionanocomposites. This required a first rheological study for assessing the effect of gelatin-CNC and CNC-CNC interactions on the colloidal state of the aqueous bionanocomposite formulations. CNC were isolated from eucalyptus pulp by sulfuric acid hydrolysis (65 wt%) at 55 °C for 30 min. Gelatin was solubilized in ultra-pure water at 85°C for 20 min and then mixed with glycerol at 20 wt.% and CNC at 0.5 wt%, 1.0 wt% and 2.5 wt%. Rotational measurements were performed to determine linear viscosity (η) of bionanocomposite solutions, which increased with increasing CNC content. At 2.5 wt% CNC, η increased by 118% regarding the neat gelatin solution, which was ascribed to percolation CNC network formation. Storage modulus (G’) and loss modulus (G″) further determined by oscillatory tests revealed that a gel-like behavior was dominant in the bionanocomposite solutions (G’ > G’’) over a broad range of temperature (20 – 85 °C), particularly at 2.5 wt% CNC. These results confirm effective interactions in the aqueous gelatin-CNC bionanocomposites that could substantially increase the physical properties of the gelatin plastics. Tensile tests are underway to confirm this hypothesis. The authors would like to thank the Fapesp (process n 2016/03080-3) for support.

Keywords: bionanocomposites, cellulose nanocrystals, gelatin, viscoelastic characterization

Procedia PDF Downloads 126

Authors: R. Hasterok, A. Betekhtin, N. Borowska, A. Braszewska-Zalewska, E. Breda, K. Chwialkowska, R. Gorkiewicz, D. Idziak, J. Kwasniewska, M. Kwasniewski, D. Siwinska, A. Wiszynska, E. Wolny

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In contrast to animals, the organisation of plant genomes at the cytomolecular level is still relatively poorly studied and understood. However, the Brachypodium genus in general and B. distachyon in particular represent exceptionally good model systems for such study. This is due not only to their highly desirable ‘model’ biological features, such as small nuclear genome, low chromosome number and complex phylogenetic relations, but also to the rapidly and continuously growing repertoire of experimental tools, such as large collections of accessions, WGS information, large insert (BAC) libraries of genomic DNA, etc. Advanced cytomolecular techniques, such as fluorescence in situ hybridisation (FISH) with evermore sophisticated probes, empowered by cutting-edge microscope and digital image acquisition and processing systems, offer unprecedented insight into chromatin organisation at various phases of the cell cycle. A good example is chromosome painting which uses pools of chromosome-specific BAC clones, and enables the tracking of individual chromosomes not only during cell division but also during interphase. This presentation outlines the present status of molecular cytogenetic analyses of plant genome structure, dynamics and evolution using B. distachyon and some of its relatives. The current projects focus on important scientific questions, such as: What mechanisms shape the karyotypes? Is the distribution of individual chromosomes within an interphase nucleus determined? Are there hot spots of structural rearrangement in Brachypodium chromosomes? Which epigenetic processes play a crucial role in B. distachyon embryo development and selective silencing of rRNA genes in Brachypodium allopolyploids? The authors acknowledge financial support from the Polish National Science Centre (grants no. 2012/04/A/NZ3/00572 and 2011/01/B/NZ3/00177)

Keywords: Brachypodium, B. distachyon, chromosome, FISH, molecular cytogenetics, nucleus, plant genome organisation

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Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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Authors: Phusit Phukamchanoad, Chutchai Ditchareon, Suwaree Yordchim

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This research was to study on background and social and cultural context of Kamchanoad community for sustainable tourism management. All data was collected through in-depth interview with village headmen, community committees, teacher, monks, Kamchanoad forest field officers and respected senior citizen above 60 years old in the community who have lived there for more than 40 years. Altogether there were 30 participants for this research. After analyzing the data, content from interview and discussion, Kamchanoad has both high land and low land in the region as well as swamps that are very capable of freshwater animals’ conservation. Kamchanoad is also good for agriculture and animal farming. 80% of Kamchanoad’s land are forest, freshwater and rice farms. Kamchanoad was officially set up as community in 1994 as “Baan Nonmuang”. Inhabitants in Kamchanoad make a living by farming based on sufficiency economy. They have rice farm, eucalyptus farm, cassava farm and rubber tree farm. Local people in Kamchanoad still believe in the myth of Srisutto Naga. They are still religious and love to preserve their traditional way of life. In order to understand how to create successful tourism business in Kamchanoad, we have to study closely on local culture and traditions. Outstanding event in Kamchanoad is the worship of Grand Srisutto, which is on the full-moon day of 6th month or Visakhabucha Day. Other big events are also celebration at the end of Buddhist lent, Naga firework, New Year celebration, Boon Mahachart, Songkran, Buddhist Lent, Boon Katin and Loy Kratong. Buddhism is the main religion in Kamchanoad. The promotion of tourism in Kamchanoad is expected to help spreading more income for this region. More infrastructures will be provided for local people as well as funding for youth support and people activities.

Keywords: social and culture area, tourism management, Kamchanoad Community, Udon Thani Province

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Authors: Young Ah Kim, Hyunsoo Kim, Eun-Jeong Yoon, Young Hee Seo, Kyungwon Lee

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Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli from food animals are considered as a reservoir for transmission of ESBL genes to human. The aim of this study is to assess the prevalence and molecular epidemiology of ESBL-producing E. coli colonization in pigs, farm workers, and farm environments to elucidate the transmission of multidrug-resistant clones from animal to human. Nineteen pig farms were enrolled across the country in Korea from August to December 2017. ESBL-producing E. coli isolates were detected in 190 pigs, 38 farm workers, and 112 sites of farm environments using ChromID ESBL (bioMerieux, Marcy l'Etoile, France), directly (stool or perirectal swab) or after enrichment (sewage). Antimicrobial susceptibility tests were done with disk diffusion methods and blaTEM, blaSHV, and blaCTX-M were detected with PCR and sequencing. The genomes of the four CTX-M-55-producing E. coli isolates from various sources in one farm were entirely sequenced to assess the relatedness of the strains. Whole genome sequencing (WGS) was performed with PacBio RS II system (Pacific Biosciences, Menlo Park, CA, USA). ESBL genotypes were 85 CTX-M-1 group (one CTX-M-3, 23 CTX-M-15, one CTX-M-28, 59 CTX-M-55, one CTX-M-69) and 60 CTX-M-9 group (41 CTX-M-14, one CTX-M-17, one CTX-M-27, 13 CTX-M-65, 4 CTX-M-102) in total 145 isolates. The rectal colonization rates were 53.2% (101/190) in pigs and 39.5% (15/38) in farm workers. In WGS, sequence types (STs) were determined as ST69 (E. coli PJFH115 isolate from a human carrier), ST457 (two E. coli isolates PJFE101 recovered from a fence and PJFA1104 from a pig) and ST5899 (E. coli PJFA173 isolate from the other pig). The four plasmids encoding CTX-M-55 (88,456 to 149, 674 base pair), whether it belonged to IncFIB or IncFIC-IncFIB type, shared IncF backbone furnishing the conjugal elements, suggesting of genes originated from same ancestor. In conclusion, the prevalence of ESBL-producing E. coli in swine farms was surprisingly high, and many of them shared common ESBL genotypes of clinical isolates such as CTX-M-14, 15, and 55 in Korea. It could spread by horizontal transfer between isolates from different reservoirs (human-animal-environment).

Keywords: Escherichia coli, extended-spectrum β-lactamase, prevalence, whole genome sequencing

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Authors: N. Nalini, Bhanu Prakash Gopularam

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The term data security refers to the degree of resistance or protection given to information from unintended or unauthorized access. The core principles of information security are the confidentiality, integrity and availability, also referred as CIA triad. Cloud computing services are classified as SaaS, IaaS and PaaS services. With cloud adoption the confidential enterprise data are moved from organization premises to untrusted public network and due to this the attack surface has increased manifold. Several cloud computing platforms like OpenStack, Eucalyptus, Amazon EC2 offer users to build and configure public, hybrid and private clouds. While the traditional encryption based on PKI infrastructure still works in cloud scenario, the management of public-private keys and trust certificates is difficult. The Identity based Public Key Cryptography (also referred as ID-PKC) overcomes this problem by using publicly identifiable information for generating the keys and works well with decentralized systems. The users can exchange information securely without having to manage any trust information. Another advantage is that access control (role based access control policy) information can be embedded into data unlike in PKI where it is handled by separate component or system. In OpenStack cloud platform the keystone service acts as identity service for authentication and authorization and has support for public key infrastructure for auto services. In this paper, we explain OpenStack security architecture and evaluate the PKI infrastructure piece for data confidentiality. We provide method to integrate ID-PKC schemes for securing data while in transit and stored and explain the key measures for safe guarding data against security attacks. The proposed approach uses JPBC crypto library for key-pair generation based on IEEE P1636.3 standard and secure communication to other cloud services.

Keywords: data confidentiality, identity based cryptography, secure communication, open stack key stone, token scoping

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Authors: Ramesh Joshi, Nisha Khatik

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Murraya koenigii (L.) Spreng locally known as “Curry patta” or “Meetha neem” belonging to the family Rutaceae that grows wildly in Southern Asia. Its aromatic leaves are commonly used as the raw material for traditional medicinal formulations in India. The leaves contain essential oil and also used as a condiment. Several monomeric and binary carbazol alkaloids present in the various plant parts. These alkaloids have been reported to possess anti-microbial, mosquitocidal, topo-isomerase inhibition and antioxidant properties. Some of the alkaloids reported in this plant have showed anti carcinogenic and anti-diabetic properties. The conventional method of propagation of this tree is limited to seeds only, which retain their viability for only a short period. Hence, a biotechnological approach might have an advantage edging over traditional breeding as well as the genetic improvement of M. koenigii within a short period. The development of a reproducible regeneration protocol is the prerequisite for ex situ conservation and micropropagation. An efficient protocol for high frequency regeneration of in vitro plants of Murraya koenigii via different explants such as- nodal segments, intermodal segments, leaf, root segments, hypocotyle, cotyledons and cotyledonary node explants is described. In the present investigation, assessment of clonal fidelity in the micropropagated plantlets of Murraya koenigii was attempted using RAPD and ISSR markers at different pathways of plant tissue culture technique. About 20 ISSR and 40 RAPD primers were used for all the samples. Genomic DNA was extracted by CTAB method. ISSR primer were found to be more suitable as compared to RAPD for the analysis of clonal fidelity of M. koenigii. The amplifications however, were finally performed using RAPD, ISSR markers owing to their better performance in terms of generation of amplification products. In RAPD primer maximum 75% polymorphism was recorded in OPU-2 series which exhibited out of 04 scorable bands, three bands were polymorphic with a band range of size 600-1500 bp. In ISSR primers the UBC 857 showed 50% polymorphism with 02 band were polymorphic of band range size between 400-1000 bp.

Keywords: genetic fidelity, Murraya koenigii, aromatic plants, ISSR primers

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Authors: M. Masroor Akhtar Khan, Moin Uddin, Lalit Varshney

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Recently, there has been a rapidly expanding interest in finding applications of natural polymers in view of value addition to agriculture. It is now being realized that radiation processing of natural polysaccharides can be beneficially utilized either to improve the existing methodologies used for processing the natural polymers or to impart value addition to agriculture by converting them into more useful form. Gamma-ray irradiation is employed to degrade and lower the molecular weight of some of the natural polysaccharides like alginates, chitosan and carrageenan into small sized oligomers. When these oligomers are applied to plants as foliar sprays, they elicit various kinds of biological and physiological activities, including promotion of plant growth, seed germination, shoot elongation, root growth, flower production, suppression of heavy metal stress, etc. Furthermore, application of these oligomers can shorten the harvesting period of various crops and help in reducing the use of insecticides and chemical fertilizers. In recent years, the oligomers of sodium alginate obtained by irradiating the latter with gamma-rays at 520 kGy dose are being employed. It was noticed that the oligomers derived from the natural polysaccharides could induce growth, photosynthetic efficiency, enzyme activities and most importantly the production of secondary metabolite in the plants like Artemisia annua, Beta vulgaris, Catharanthus roseus, Chrysopogon zizanioides, Cymbopogon flexuosus, Eucalyptus citriodora, Foeniculum vulgare, Geranium sp., Mentha arvensis, Mentha citrata, Mentha piperita, Mentha virdis, Papaver somniferum and Trigonella foenum-graecum. As a result of the application of these oligomers, the yield and/or contents of the active constituents of the aforesaid plants were significantly enhanced. The productivity, as well as quality of medicinal and aromatic plants, may be ameliorated by this novel technique in an economical way as a very little quantity of these irradiated (depolymerised) polysaccharides is needed. Further, this is a very safe technique, as we did not expose the plants directly to radiation. The radiation was used to depolymerize the polysaccharides into oligomers.

Keywords: essential oil, medicinal and aromatic plants, plant production, radiation processed polysaccharides, active constituents

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Authors: Hanamariam Mekonnen

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Ethiopian farmers traditionally produce charcoal under several kilns on cultivated land: particularly in Kasiry micro-watershed Fagita Lekoma district of Northwestern Ethiopia. However, the effects of such soil heating and remnants of charcoal leftover on soils have not been adequately documented. Hence, this study tried to quantify the effects of such kiln sites on selected soil properties and wheat crop performance. Soils from four kiln sites were thus purposively sampled at depths of 0-20 cm, 20-40 cm and 40-60 cm and were compared with the respective soil layers of none-kiln sites from similar adjacent fields. While soil moisture content was sampled at kiln and none-kiln site in wet and dry seasons from each depth. In addition, a pot experiment was conducted using two sources of biochar (Acacia decurrens and Eucalyptus Camaldulensis) with four rates (0, 10, 20, and 40 t/ha) and compared with crops grown from soils of 1kiln sites without biochar application laid out in a CRD with three replications. The data were analyzed using SAS software Version 9.4.The result revealed notable variations of kiln site soils and along soil depth. The appreciable increased (p<0.05) soil pH (5.5 to 5.74), organic carbon (3.89 to 4.27%), TN (0.30 to 0.32%), CEC (32.59 to 35.23 cmolckg-1), Ca (6.44 to 7.9 cmolckg-1), Mg (4.48 to 5.46 cmolckg-1), and significantly (p<0.01) Av. P (30.25 to 46.4 ppm) and K (2.11 to 2.82 cmolckg-1) were recorded from the none-kiln to kiln soils, respectively. On the other hand, ex. acidity and aluminum, available Fe and Mn were reduced from 2.20 to 1.54, 1.95 to 1.31 cmolckg-1 and 57.46 to 41.40 and 5.65 to 3.86 ppm, respectively, from the control to the kiln. Soil texture was significantly affected by soil heating and along soil depth. The sand content was (p<0.05) varied between the value of 23% to 29% from none-kiln to kiln site, and clay content was (p<0.01) increased from 0-20 cm (32%) soil depth to 40-60 cm (43%) deeper soil. Significantly (p<0.05) higher Soil moisture content was recorded at none-kiln site (45.85%) compared to kiln (40.44%) in wet season, whereas in dry season, lower moisture content was revealed at kiln site (26%) compared to none-kiln (30.7%). As wet to dry season, soil moisture was decreased from 43% to 28% respectively. Bulk density (P<0.01) varied between 0.88 to 0.94 gcm-3 from control to kiln in dry season. Similarly, the value of soil pH (6.10), Av. P (58.12), exchangeable bases (Ca (9.83), Mg (6.19) and K (3.67)) were (p<0.01) higher at the 0-20 cm soil depth as compared to the deeper soils, the result of soil moisture (30 to 42%) and CEC (31 to 36 cmolckg-1) increased down the soil profile. After wheat harvest, soil pH, Av. P, CEC, and exchangeable bases (Mg, K and Na) were significantly higher in the kiln soil, while soil moisture and OC increased by the applied biochar of 20 and 40 ton/ha. High yield 2.28 gpot-1 (p<0.01) was recorded in kiln soil, growth parameters of wheat were significantly increased with increasing biochar rates.

Keywords: biochar, kasiry micro-watershed, kiln site, none-kiln site, soil properties

Procedia PDF Downloads 37

Authors: Japhet N. Mwambusi

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High tropical forests deforestation for solid wood furniture industry is among of climate change contributing agents. This pressure indirectly is caused by furniture joints failure due to poor gluing technology based on improper use of different glues to different wood species which lead to low quality and weak wood-glue joints. This study was carried in order to run performance tests of wood glues on different wood species used in wood workshops: Morogoro Tanzania whereby three popular wood species of C. lusitanica, T. glandis and E. maidenii were tested against five glues of Woodfix, Bullbond, Ponal, Fevicol and Coral found in the market. The findings were necessary on developing a guideline for proper glue selection for a particular wood species joining. Random sampling was employed to interview carpenters while conducting a survey on the background of carpenters like their education level and to determine factors that influence their glues choice. Monsanto Tensiometer was used to determine bonding strength of identified wood glues to different wood species in use under British Standard of testing wood shear strength (BS EN 205) procedures. Data obtained from interviewing carpenters were analyzed through Statistical Package of Social Science software (SPSS) to allow the comparison of different data while laboratory data were compiled, related and compared by the use of MS Excel worksheet software as well as Analysis of Variance (ANOVA). Results revealed that among all five wood glues tested in the laboratory to three different wood species, Coral performed much better with the average shear strength 4.18 N/mm², 3.23 N/mm² and 5.42 N/mm² for Cypress, Teak and Eucalyptus respectively. This displays that for a strong joint to be formed to all tree wood species for soft wood and hard wood, Coral has a first priority in use. The developed table of guideline from this research can be useful to carpenters on proper glue selection to a particular wood species so as to meet glue-bond strength. This will secure furniture market as well as reduce pressure to the forests for furniture production because of the strong existing furniture due to their strong joints. Indeed, this can be a good strategy on reducing climate change speed in tropics which result from high deforestation of trees for furniture production.

Keywords: climate change, deforestation, gluing technology, joint failure, wood-glue, wood species

Procedia PDF Downloads 213

Authors: Kubilay Kurtulus Bastas

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Bacterial diseases are very destructive and cause economic losses on pears. Promising plant extracts for the management of plant diseases are environmentally safe, long-lasting and extracts of certain plants contain alkaloids, tannins, quinones, coumarins, phenolic compounds, and phytoalexins. In this study, bacteria were isolated from different parts of pear exhibiting characteristic symptoms of bacterial diseases from the Central Anatolia, Turkey. Pathogenic bacteria were identified by morphological, physiological, biochemical and molecular methods as fire blight (Erwinia amylovora (39%)), bacterial blossom blast and blister bark (Pseudomonas syringae pv. syringae (22%)), crown gall (Rhizobium radiobacter (1%)) from different pear cultivars, and determined virulence levels of the pathogens with pathogenicity tests. The air-dried 25 plant material was ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration (MIC) values were determined by using modified disc diffusion method at five different concentrations and streptomycin sulphate was used as control chemical. Bacterial suspensions were prepared as 108 CFU ml⁻¹ densities and 100 µl bacterial suspensions were spread to TSA medium. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the test organisms. Among the tested plants, Origanum vulgare, Hedera helix, Satureja hortensis, Rhus coriaria, Eucalyptus globulus, Rosmarinus officinalis, Ocimum basilicum, Salvia officinalis, Cuminum cyminum and Thymus vulgaris showed a good antibacterial activity and they inhibited the growth of the pathogens with inhibition zone diameter ranging from 7 to 27 mm at 20% (w/v) in absolute methanol in vitro conditions. In vivo, the highest efficacy was determined as 27% on reducing tumor formation of R. radiobacter, and 48% and 41% on reducing shoot blight of E. amylovora and P. s. pv. syringae on pear seedlings, respectively. Obtaining data indicated that some plant extracts may be used against the bacterial diseases on pome fruits within sustainable and organic management programs.

Keywords: bacteria, eco-friendly management, organic, pear, plant extract

Procedia PDF Downloads 294

Authors: Michael Russelle S. Alvarez, Noel S. Quiming, Francisco M. Heralde

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This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.

Keywords: larvicidal activity screening, partial purification, dose-response assay, capsicum frutescens

Procedia PDF Downloads 302

Authors: Mehwish Jamil Noor

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Morphological features of pollen (sculpturing) were useful for identification of different floral taxa. In this study 49 pollen grains, types belonging to 25 families were studied using Scanning Electron Microscope. Shape and sculpturing of pollen ranging from Psilate, scabrate to reticulate, bireticulate and echinolophate. Honey pollen was identified using morphological features, number and arrangement of pore and colpi, size and shape. It presents the first attempt from Pakistan involving extraction of pollen from honey, its identification and taxonomic analysis. Among pollen studied diversity in shape and sculpturing has been observed ranging from Psilate, scabrate to reticulate to bireticulate and echinolophate condition. Pollen has been identified with the help of morphological feature, number and arrangement of pore and colpi, size and shape, reference slides, light microscopic data and previous literature have been consulted for pollen identification. Pollen of closely related species resemble each other therefore pollen identification of airborne and honey pollen is not possible till species level. Survey of flora was carried in parallel to keep the record about the allergenic and melliferous preference of specific sites through surveys and interviews. Their pollination season and geographical distribution were recorded. Two hundred and five including wild and cultivated taxa were identified belonging to sixty-seven families. Major bee attracting wild shrub and trees includes Justicia adhatoda, Acacia nilotica, Ziziphus jujuba, Taraxicum officinalis, Artemisia dubia, Casuarina sp., Ulmus sp., Broussonetia papyrifera, Cupressus sp. or Pinus roxburghii etc. Cultivated crops like Pennisetum typhoides, Nigella sativa, Triticum sativum along with fruit trees of Pyrus, Prunus, Eryobotria, Citrus etc. are popular melliferous floras. Exotic/ introduced species like Eucalyptus or Parthenium hysterophorus, are also frequently visited by bees indicating the significance of those plants in the honey industry. It is concluded that different microscopic analysis techniques give more clear and authentic pictures of and melliferous pollen identification which is well supported by the floral calendar. The diversity of pollen are observed in case of melliferous pollen, and most of the windborne pollen were found less sculptured or psilate expressing the adaptation to the specific mode of pollination. Pollen morphology and sculpturing would serve as a reference for future studies.

Keywords: pollen, allergenic flora, sem, pollen key, Scanning Electron Microscopy (SEM)

Procedia PDF Downloads 169

Authors: Stephen Osakue Amiandamhen, Schwaller Andreas, Martina Meincken, Luvuyo Tyhoda

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Invasive alien tree species are currently being cleared in South Africa as a result of the forest and water imbalances. These species grow wildly constituting about 40% of total forest area. They compete with the ecosystem for natural resources and are considered as ecosystem engineers by rapidly changing disturbance regimes. As such, they are harvested for commercial uses but much of it is wasted because of their form and structure. The waste is being sold to local communities as fuel wood. These species can be considered as potential feedstock for the production of phosphate bonded composites. The presence of bark in wood-based composites leads to undesirable properties, and debarking as an option can be cost implicative. This study investigates the potentials of these invasive species processed without debarking on some fundamental properties of wood-based panels. Some invasive alien tree species were collected from EC Biomass, Port Elizabeth, South Africa. They include Acacia mearnsii (Black wattle), A. longifolia (Long-leaved wattle), A. cyclops (Red-eyed wattle), A. saligna (Golden-wreath wattle) and Eucalyptus globulus (Blue gum). The logs were chipped as received. The chips were hammer-milled and screened through a 1 mm sieve. The wood particles were conditioned and the quantity of bark in the wood was determined. The binding matrix was prepared using a reactive magnesia, phosphoric acid and class S fly ash. The materials were mixed and poured into a metallic mould. The composite within the mould was compressed at room temperature at a pressure of 200 KPa. After initial setting which took about 5 minutes, the composite board was demoulded and air-cured for 72 h. The cured product was thereafter conditioned at 20°C and 70% relative humidity for 48 h. Test of physical and strength properties were conducted on the composite boards. The effect of binder formulation and fly ash content on the properties of the boards was studied using fitted response surface technology, according to a central composite experimental design (CCD) at a fixed wood loading of 75% (w/w) of total inorganic contents. The results showed that phosphate/magnesia ratio of 3:1 and fly ash content of 10% was required to obtain a product of good properties and sufficient strength for intended applications. The proposed products can be used for ceilings, partitioning and insulating wall panels.

Keywords: invasive alien tree species, phosphate bonded composites, physical properties, strength

Procedia PDF Downloads 263

Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson

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Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.

Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model

Procedia PDF Downloads 278

Authors: Alejandro Ruiz-Olivares, M. del Carmen González-Chávez, Rogelio Carrillo-González, Martha Reyes-Ramos, Javier Suárez Espinosa

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Use of automobiles has increased and proportionally, the demand for batteries to impulse them. When the device is aged, all the battery materials are reused through lead acid battery recycling (LABR). Importation of used lead acid batteries in Mexico has increased in the last years since many recycling factories have been settled in the country. Inadequate disposal of lead-acid battery recycling (LABR) wastes left soil severely polluted with Pb, Cu, and salts (Na+, SO2− 4, PO3− 4). Soil organic amendments may contribute with essential nutrients and sequester (scavenger compounds) metals to allow plant establishment. The objective of this research was to revegetate a former lead-acid battery recycling site aided with organic amendments. Seven tree species (Acacia farnesiana, Casuarina equisetifolia, Cupressus lusitanica, Eucalyptus obliqua, Fraxinus excelsior, Prosopis laevigata and Pinus greggii) and two organic amendments (vermicompost and vermicompost + sawdust mixture) were tested for phytoremediation of a defunct LABR site. Plants were irrigated during the dry season. Monitoring of the soils was carried out during the experiment: Available metals, salts concentrations and their spatial pattern in soil were analyzed. Plant species and amendments were compared through analysis of covariance and longitudinal analysis. High concentrations of extractable (DTPA-TEA-CaCl₂) metals (up to 15,685 mg kg⁻¹ and 478 mg kg⁻¹ for Pb and Cu) and soluble salts (292 mg kg-1 and 23,578 mg kg-1 for PO3− 4and SO2− 4) were found in the soil after three and six months of setting up the experiment. Lead and Cu concentrations were depleted in the rhizosphere after amendments addition. Spatial pattern of PO3− 4, SO2− 4 and DTPA-extractable Pb and Cu changed slightly through time. In spite of extreme soil conditions the plant species planted: A. farnesiana, E. obliqua, C. equisetifolia and F. excelsior had 100% of survival. Available metals and salts differently affected each species. In addition, negative effect on growth due to Pb accumulated in shoots was observed only in C. lusitanica. Many specimens accumulated high concentrations of Pb ( > 1000 mg kg-1) in shoots. C. equisetifolia and C. lusitanica had the best rate of growth. Based on the results, all the evaluated species may be useful for revegetation of Pb-polluted soils. Besides their use in phytoremediation, some ecosystem services can be obtained from the woodland such as encourage wildlife, wood production, and carbon sequestration. Further research should be conducted to analyze these services.

Keywords: heavy metals, inadequate disposal, organic amendments, phytoremediation with trees

Procedia PDF Downloads 257

Authors: Prabhat Kashyap, Krishan Kumar

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Delhi, the capital of India, is one of the most populated and polluted city among the world. In terms of air quality, Delhi’s air is degrading day by day & becomes worst of any major city in the world. The role of biogenic volatile organic compounds (BVOCs) is not much studied in cities like Delhi as a culprit for degraded air quality. They not only play a critical role in rural areas but also determine the atmospheric chemistry of urban areas as well. Particularly, Isoprene (2-methyl 1,3-butadiene, C5H8) is the single largest emitted compound among other BVOCs globally, that influence the tropospheric ozone chemistry in urban environment as the ozone forming potential of isoprene is very high. It is mainly emitted by vegetation & a small but significant portion is also released by vehicular exhaust of petrol operated vehicles. This study investigates the spatial and temporal variations of quantitative measurements of isoprene emissions along with different traffic tracers in 2 different seasons (post-monsoon & winter) at four different locations of Delhi. For the quantification of anthropogenic and biogenic isoprene, two sites from traffic intersections (Punjabi Bagh & CRRI) and two sites from vegetative locations (JNU & Yamuna Biodiversity Park) were selected in the vicinity of isoprene emitting tree species like Ficus religiosa, Dalbergia sissoo, Eucalyptus species etc. The concentrations of traffic tracers like benzene, toluene were also determined & their robust ratios with isoprene were used to differentiate anthropogenic isoprene with biogenic portion at each site. The ozone forming potential (OFP) of all selected species along with isoprene was also estimated. For collection of intra-day samples (3 times a day) in each season, a pre-conditioned fenceline monitoring (FLM) carbopack X thermal desorption tubes were used and further analysis was done with Gas chromatography attached with mass spectrometry (GC-MS). The results of the study proposed that the ambient air isoprene is always higher in post-monsoon season as compared to winter season at all the sites because of high temperature & intense sunlight. The maximum isoprene emission flux was always observed during afternoon hours in both seasons at all sites. The maximum isoprene concentration was found to be 13.95 ppbv at Biodiversity Park during afternoon time in post monsoon season while the lower concentration was observed as low as 0.07 ppbv at the same location during morning hours in winter season. OFP of isoprene at vegetation sites is very high during post-monsoon because of high concentrations. However, OFP for other traffic tracers were high during winter seasons & at traffic locations. Furthermore, high correlation between isoprene emissions with traffic volume at traffic sites revealed that a noteworthy share of its emission also originates from road traffic.

Keywords: biogenic VOCs, isoprene emission, anthropogenic isoprene, urban vegetation

Procedia PDF Downloads 96

Authors: K. J. Sannapapamma, H. Malligawad Lokanath, Sakeena Naikwadi

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Eco-friendly textiles are gaining importance among the consumers and textile manufacturers in the healthcare sector due to increased environmental pollution which leads to several health and environmental hazards. Hence, the research was designed to cultivate and develop the organic cotton knit, to prepare and characterize the Vetiver oil microcapsules for textile finishing and to access the wash durability of finished knits. The cotton SAHANA variety grown under organic production systems was processed and spun into 30 single yarn dyed with four natural colorants (Arecanut slurry, Eucalyptus leaves, Pomegranate rind and Indigo) and eco dyed yarn was further used for development of single jersy knitted fabric. Vetiveria zizanioides is an aromatic grass which is being traditionally used in medicine and perfumery. Vetiver essential oil was used for preparation of microcapsules by interfacial polymerization technique subjected to Gas Chromatography Mass Spectrometry (GCMS), Fourier Transform Infrared Spectroscopy (FTIR), Thermo Gravimetric Analyzer (TGA) and Scanning Electron Microscope (SEM) for characterization of microcapsules. The knitted fabric was finished with vetiver oil microcapsules by exhaust and pad dry cure methods. The finished organic knit was assessed for laundering on antimicrobial efficiency and aroma intensity. GCMS spectral analysis showed that, diethyl phthalate (28%) was the major compound found in vetiver oil followed by isoaromadendrene epoxide (7.72%), beta-vetivenene (6.92%), solavetivone (5.58%), aromadenderene, azulene and khusimol. Bioassay explained that, the vetiver oil and diluted vetiver oil possessed greater zone of inhibition against S. aureus and E. coli than the coconut oil. FTRI spectra of vetiver oil and microcapsules possessed similar peaks viz., C-H, C=C & C꞊O stretching and additionally oil microcapsules possessed the peak of 3331.24 cm^-1 at 91.14 transmittance was attributed to N-H stretches. TGA of oil microcapsules revealed that, there was a minimum weight loss (5.835%) recorded at 467.09°C compared to vetiver oil i.e., -3.026% at the temperature of 396.24°C. The shape of the microcapsules was regular and round, some were spherical in shape and few were rounded by small aggregates. Irrespective of methods of application, organic cotton knits finished with microcapsules by pad dry cure method showed maximum zone of inhibition compared to knits finished by exhaust method against S. aureus and E. coli. The antimicrobial activity of the finished samples was subjected to multiple washing which indicated that knits finished with pad dry cure method showed a zone of inhibition even after 20^th wash and better aroma retention compared to knits finished with the exhaust method of application. Further, the group of respondents rated that the 5^th washed samples had the greater aroma intensity in both the methods than the other samples. Thus, the vetiver microencapsulated organic cotton knits are free from hazardous chemicals and have multi-functional properties that can be suitable for medical and healthcare textiles.

Keywords: exhaust and pad dry cure finishing, interfacial polymerization, organic cotton knits, vetiver oil microcapsules

Procedia PDF Downloads 237

Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri

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The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.

Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq

Procedia PDF Downloads 72

Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

Procedia PDF Downloads 126

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

Abstract:

Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

Procedia PDF Downloads 51

Authors: Aswathaman Vijayan, Manish Jha, Ullas Theertha

Abstract:

Climate change and Global warming have become an issue for both developed and developing countries and perhaps the biggest threat to the environment. We at ITC Limited believe that a company’s performance must be measured by its Triple Bottom Line contribution to building economic, social and environmental capital. This Triple Bottom Line strategy focuses on - Embedding sustainability in business practices, Investing in social development and Adopting a low carbon growth path with a cleaner environment approach. The Agri Business Division - ILTD operates in the tobacco crop growing regions of Andhra Pradesh and Karnataka province of India. The Agri value chain of the company comprises of two distinct phases: First phase is Agricultural operations undertaken by ITC trained farmers and the second phase is Industrial operations which include marketing and processing of the agricultural produce. This research work covers the Greenhouse Gas (GHG) management strategy of ITC in the Agricultural operations undertaken by the farmers. The agriculture sector adds considerably to global GHG emissions through the use of carbon-based energies, use of fertilizers and other farming operations such as ploughing. In order to minimize the impact of farming operations on the environment, ITC has a taken a big leap in implementing system and process in reducing the GHG impact in farm value chain by partnering with the farming community. The company has undertaken a unique three-pronged approach for GHG management at the farm value chain: 1) GHG inventory at farm value chain: Different sources of GHG emission in the farm value chain were identified and quantified for the baseline year, as per the IPCC guidelines for greenhouse gas inventories. The major sources of emission identified are - emission due to nitrogenous fertilizer application during seedling production and main-field; emission due to diesel usage for farm machinery; emission due to fuel consumption and due to burning of crop residues. 2) Identification and implementation of technologies to reduce GHG emission: Various methodologies and technologies were identified for each GHG emission source and implemented at farm level. The identified methodologies are – reducing the consumption of chemical fertilizer usage at the farm through site-specific nutrient recommendation; Usage of sharp shovel for land preparation to reduce diesel consumption; implementation of energy conservation technologies to reduce fuel requirement and avoiding burning of crop residue by incorporation in the main field. These identified methodologies were implemented at farm level, and the GHG emission was quantified to understand the reduction in GHG emission. 3) Social and farm forestry for CO2 sequestration: In addition, the company encouraged social and farm forestry in the waste lands to convert it into green cover. The plantations are carried out with fast growing trees viz., Eucalyptus, Casuarina, and Subabul at the rate of 10,000 Ha of land per year. The above approach minimized considerable amount of GHG emission at the farm value chain benefiting farmers, community, and environment at a whole. In addition, the CO₂ stock created by social and farm forestry program has made the farm value chain to become environment-friendly.

Keywords: CO₂ sequestration, farm value chain, greenhouse gas, ITC limited

Procedia PDF Downloads 270