Search results for: alien species
121 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics
Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh
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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity
Procedia PDF Downloads 144120 Population Diversity of Dalmatian Pyrethrum Based on Pyrethrin Content and Composition
Authors: Filip Varga, Nina Jeran, Martina Biosic, Zlatko Satovic, Martina Grdisa
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Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.), a species endemic to the eastern Adriatic coastline, is the source of natural insecticide pyrethrin. Pyrethrin is a mixture of six compounds (pyrethrin I and II, cinerin I and II, jasmolin I and II) that exhibits high insecticidal activity with no detrimental effects to the environment. A recently optimized matrix-solid phase dispersion method (MSPD), using florisil as the sorbent, acetone-ethyl acetate (1:1, v/v) as the elution solvent, and sodium sulfate anhydrous as the drying agent was utilized to extract the pyrethrins from 10 wild populations (20 individuals per population) distributed along the Croatian coast. All six components in the extracts were qualitatively and quantitatively determined by high-performance liquid chromatography with a diode array detector (HPLC-DAD). Pearson’s correlation index was calculated between pyrethrin compounds, and differences between the populations using the analysis of variance were tested. Additionally, the correlation of each pyrethrin component with spatio-ecological variables (bioclimate, soil properties, elevation, solar radiation, and distance from the coastline) was calculated. Total pyrethrin content ranged from 0.10% to 1.35% of dry flower weight, averaging 0.58% across all individuals. Analysis of variance revealed significant differences between populations based on all six pyrethrin compounds and total pyrethrin content. On average, the lowest total pyrethrin content was found in the population from Pelješac peninsula (0.22% of dry flower weight) in which total pyrethrin content lower than 0.18% was detected in 55% of the individuals. The highest average total pyrethrin content was observed in the population from island Zlarin (0.87% of dry flower weight), in which total pyrethrin content higher than 1.00% was recorded in only 30% of the individuals. Pyrethrin I/pyrethrin II ratio as a measure of extract quality ranged from 0.21 (population from the island Čiovo) to 5.88 (population from island Mali Lošinj) with an average of 1.77 across all individuals. By far, the lowest quality of extracts was found in the population from Mt. Biokovo (pyrethrin I/II ratio lower than 0.72 in 40% of individuals) due to the high pyrethrin II content typical for this population. Pearson’s correlation index revealed a highly significant positive correlation between pyrethrin I content and total pyrethrin content and a strong negative correlation between pyrethrin I and pyrethrin II. The results of this research clearly indicate high intra- and interpopulation diversity of Dalmatian pyrethrum with regards to pyrethrin content and composition. The information obtained has potential use in plant genetic resources conservation and biodiversity monitoring. Possibly the largest potential lies in designing breeding programs aimed at increasing pyrethrin content in commercial breeding lines and reintroduction in agriculture in Croatia. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).Keywords: Dalmatian pyrethrum, HPLC, MSPD, pyrethrin
Procedia PDF Downloads 142119 Effect of Time on Stream on the Performances of Plasma Assisted Fe-Doped Cryptomelanes in Trichloroethylene (TCE) Oxidation
Authors: Sharmin Sultana, Nicolas Nuns, Pardis Simon, Jean-Marc Giraudon, Jean-Francois Lamonior, Nathalie D. Geyter, Rino Morent
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Environmental issues, especially air pollution, have become a huge concern of environmental legislation as a consequence of growing awareness in our global world. In this regard, control of volatile organic compounds (VOCs) emission has become an important issue due to their potential toxicity, carcinogenicity, and mutagenicity. The research of innovative technologies for VOC abatement is stimulated to accommodate the new stringent standards in terms of VOC emission. One emerging strategy is the coupling of 2 existing complementary technologies, namely here non-thermal plasma (NTP) and heterogeneous catalysis, to get a more efficient process for VOC removal in air. The objective of this current work is to investigate the abatement of trichloroethylene (TCE-highly toxic chlorinated VOC) from moist air (RH=15%) as a function of time by combined use of multi-pin-to-plate negative DC corona/glow discharge with Fe-doped cryptomelanes catalyst downstream i.e. post plasma-catalysis (PPC) process. For catalyst alone case, experiments reveal that, initially, Fe doped cryptomelane (regardless the mode of Fe incorporation by co-precipitation (Fe-K-OMS-2)/ impregnation (Fe/K-OMS-2)) exhibits excellent activity to decompose TCE compared to cryptomelane (K-OMS-2) itself. A maximum obtained value of TCE abatement after 6 min is as follows: Fe-KOMS-2 (73.3%) > Fe/KOMS-2 (48.5) > KOMS-2 (22.6%). However, with prolonged operation time, whatever the catalyst under concern, the abatement of TCE decreases. After 111 min time of exposure, the catalysts can be ranked as follows: Fe/KOMS-2 (11%) < K-OMS-2 (12.3%) < Fe-KOMS-2 (14.5%). Clearly, this phenomenon indicates catalyst deactivation either by chlorination or by blocking the active sites. Remarkably, in PPC configuration (energy density = 60 J/L, catalyst temperature = 150°C), experiments reveal an enhanced performance towards TCE removal regardless the type of catalyst. After 6 min time on stream, the TCE removal efficiency amount as follows: K-OMS-2 (60%) < Fe/K-OMS-2 (79%) < Fe-K-OMS-2 (99.3%). The enhanced performances over Fe-K-OMS-2 catalyst are attributed to its high surface oxygen mobility and structural defects leading to high O₃ decomposition efficiency to give active species able to oxidize the plasma processed hazardous\by-products and the possibly remaining VOC into CO₂. Moreover, both undoped and doped catalysts remain strongly capable to abate TCE with time on stream. The TCE removal efficiencies of the PPC processes with Fe/KOMS-2 and KOMS-2 catalysts are not affected by time on stream indicating an excellent catalyst stability. When using the Fe-K-OMS-2 as catalyst, TCE abatement slightly reduces with time on stream. However, it is noteworthy to stress that still a constant abatement of 83% is observed during at least 30 minutes. These results prove that the combination of NTP with catalysts not only increases the catalytic activity but also allows to avoid, to some extent, the poisoning of catalytic sites resulting in an enhanced catalyst stability. In order to better understand the different surface processes occurring in the course of the total TCE oxidation in PPC experiments, a detailed X-ray Photoelectron Spectroscopy (XPS) and Time of Flight-Secondary Ion Mass Spectrometry (ToF-SIMS) study on the fresh and used catalysts is in progress.Keywords: Fe doped cryptomelane, non-thermal plasma, plasma-catalysis, stability, trichloroethylene
Procedia PDF Downloads 208118 Photophysics and Torsional Dynamics of Thioflavin T in Deep Eutectic Solvents
Authors: Rajesh Kumar Gautam, Debabrata Seth
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Thioflavin-T (ThT) play a key role of an important biologically active fluorescent sensor for amyloid fibrils. ThT molecule has been developed a method to detect the analysis of different type of diseases such as neurodegenerative disorders, Alzheimer’s, Parkinson’s, and type II diabetes. ThT was used as a fluorescent marker to detect the formation of amyloid fibril. In the presence of amyloid fibril, ThT becomes highly fluorescent. ThT undergoes twisting motion around C-C bonds of the two adjacent benzothiazole and dimethylaniline aromatic rings, which is predominantly affected by the micro-viscosity of the local environment. The present study articulates photophysics and torsional dynamics of biologically active molecule ThT in the presence of deep-eutectic solvents (DESs). DESs are environment-friendly, low cost and biodegradable alternatives to the ionic liquids. DES resembles ionic liquids, but the constituents of a DES include a hydrogen bond donor and acceptor species, in addition to ions. Due to the presence of the H-bonding network within a DES, it exhibits structural heterogeneity. Herein, we have prepared two different DESs by mixing urea with choline chloride and N, N-diethyl ethanol ammonium chloride at ~ 340 K. It was reported that deep eutectic mixture of choline chloride with urea gave a liquid with a freezing point of 12°C. We have experimented by taking two different concentrations of ThT. It was observed that at higher concentration of ThT (50 µM) it forms aggregates in DES. The photophysics of ThT as a function of temperature have been explored by using steady-state, and picoseconds time-resolved fluorescence emission spectroscopic techniques. From the spectroscopic analysis, we have observed that with rising temperature the fluorescence quantum yields and lifetime values of ThT molecule gradually decreases; this is the cumulative effect of thermal quenching and increase in the rate of the torsional rate constant. The fluorescence quantum yield and fluorescence lifetime decay values were always higher for DES-II (urea & N, N-diethyl ethanol ammonium chloride) than those for DES-I (urea & choline chloride). This was mainly due to the presence of structural heterogeneity of the medium. This was further confirmed by comparison with the activation energy of viscous flow with the activation energy of non-radiative decay. ThT molecule in less viscous media undergoes a very fast twisting process and leads to deactivation from the photoexcited state. In this system, the torsional motion increases with increasing temperature. We have concluded that beside bulk viscosity of the media, structural heterogeneity of the medium play crucial role to guide the photophysics of ThT in DESs. The analysis of the experimental data was carried out in the temperature range 288 ≤ T = 333K. The present articulate is to obtain an insight into the DESs as media for studying various photophysical processes of amyloid fibrils sensing molecule of ThT.Keywords: deep eutectic solvent, photophysics, Thioflavin T, the torsional rate constant
Procedia PDF Downloads 162117 Disease Control of Rice Blast Caused by Pyricularia Oryzae Cavara Using Novel Chitosan-based Agronanofungicides
Authors: Abdulaziz Bashir Kutawa, Khairulmazmi Ahmad, Mohd Zobir Hussein, Asgar Ali, Mohd Aswad Abdul Wahab, Amara Rafi, Mahesh Tiran Gunasena, Muhammad Ziaur Rahman, Md. Imam Hossain, Syazwan Afif Mohd Zobir
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Rice is a cereal crop and belongs to the family Poaceae, it was domesticated in southern China and North-Eastern India around 8000 years ago, and it’s the staple nourishment for over half of the total world’s population. Rice production worldwide is affected by different abiotic and biotic stresses. Diseases are important challenges for the production of rice, among all the diseases in rice plants, the most severe and common disease is the rice blast. Worldwide, it is one of the most damaging diseases affecting rice cultivation, the disease is caused by the non-obligate filamentous ascomycete fungus called Magnaporthe grisae or Pyricularia oryzae Cav. Nanotechnology is a new idea to improve agriculture by combating the diseases of plants, as nanoparticles were found to possess an inhibitory effect on different species of fungi. This work aimed to develop and determine the efficacy of agronanofungicides, and commercial fungicides (in-vitro and in-vivo). The agronanofungicides were developed using ionic gelation methods. In-vitro antifungal activity of the synthesized agronanofungicides was evaluated against P. oryzae using the poisoned medium technique. The potato dextrose agar (PDA) was amended in several concentrations; 0.001, 0.005, 0.01, 0.025, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, and 0.35 ppm for the agronanofungicides. Medium with the only solvent served as a control. Mycelial growth was recorded every day, and the percentage inhibition of radial growth (PIRG) was also calculated. Based on the results of the zone of inhibition, the chitosan-hexaconazole agronanofungicide (2g/mL) was the most effective fungicide to inhibit the growth of the fungus with 100% inhibition at 0.2, 0.25, 0.30, and 0.35 ppm, respectively. The least were found to be propiconazole and basamid fungicides with 100% inhibition only at 100 ppm. In terms of the glasshouse results, the chitosan-hexaconazole-dazomet agronanofungicide (CHDEN) treatment (2.5g/L) was found to be the most effective fungicide to reduce the intensity of the disease with a disease severity index (DSI) of 19.80%, protection index (PI) of 82.26%, lesion length of 1.63cm, disease reduction (DR) of 80.20%, and AUDPC (390.60 Unit2). The least effective fungicide was found to be ANV with a disease severity index (45.60%), protection index (45.24%), lesion length (3.83 cm), disease reduction (54.40%), and AUDPC (1205.75 Unit2). The negative control did not show any symptoms during the glasshouse assay, while the untreated control treatment exhibited severe symptoms of the disease with a DSI value of 64.38%, lesion length of 5.20 cm, and AUDPC value of 2201.85 Unit2, respectively. The treatments of agronanofungicides have enhanced the yield significantly with CHDEN having 239.00 while the healthy control had 113.67 for the number of grains per panicle. The use of CHEN and CHDEN will help immensely in reducing the severity of rice blast in the fields, and this will increase the yield and profit of the farmers that produced rice.Keywords: chitosan, dazomet, disease severity, efficacy, and blast disease
Procedia PDF Downloads 87116 Glucose Measurement in Response to Environmental and Physiological Challenges: Towards a Non-Invasive Approach to Study Stress in Fishes
Authors: Tomas Makaras, Julija Razumienė, Vidutė Gurevičienė, Gintarė Sauliutė, Milda Stankevičiūtė
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Stress responses represent animal’s natural reactions to various challenging conditions and could be used as a welfare indicator. Regardless of the wide use of glucose measurements in stress evaluation, there are some inconsistencies in its acceptance as a stress marker, especially when it comes to comparison with non-invasive cortisol measurements in the fish challenging stress. To meet the challenge and to test the reliability and applicability of glucose measurement in practice, in this study, different environmental/anthropogenic exposure scenarios were simulated to provoke chemical-induced stress in fish (14-days exposure to landfill leachate) followed by a 14-days stress recovery period and under the cumulative effect of leachate fish subsequently exposed to pathogenic oomycetes (Saprolegnia parasitica) to represent a possible infection in fish. It is endemic to all freshwater habitats worldwide and is partly responsible for the decline of natural freshwater fish populations. Brown trout (Salmo trutta fario) and sea trout (Salmo trutta trutta) juveniles were chosen because of a large amount of literature on physiological stress responses in these species was known. Glucose content in fish by applying invasive and non-invasive glucose measurement procedures in different test mediums such as fish blood, gill tissues and fish-holding water were analysed. The results indicated that the quantity of glucose released in the holding water of stressed fish increased considerably (approx. 3.5- to 8-fold) and remained substantially higher (approx. 2- to 4-fold) throughout the stress recovery period than the control level suggesting that fish did not recover from chemical-induced stress. The circulating levels of glucose in blood and gills decreased over time in fish exposed to different stressors. However, the gill glucose level in fish showed a decrease similar to the control levels measured at the same time points, which was found to be insignificant. The data analysis showed that concentrations of β-D glucose measured in gills of fish treated with S. parasitica differed significantly from the control recovery, but did not differ from the leachate recovery group showing that S. parasitica presence in water had no additive effects. In contrast, a positive correlation between blood and gills glucose were determined. Parallel trends in blood and water glucose changes suggest that water glucose measurement has much potency in predicting stress. This study demonstrated that measuring β-D-glucose in fish-holding water is not stressful as it involves no handling and manipulation of an organism and has critical technical advantages concerning current (invasive) methods, mainly using blood samples or specific tissues. The quantification of glucose could be essential for studies examining the stress physiology/aquaculture studies interested in the assessment or long-term monitoring of fish health.Keywords: brown trout, landfill leachate, sea trout, pathogenic oomycetes, β-D-glucose
Procedia PDF Downloads 173115 Clinicomycological Pattern of Superficial Fungal Infections among Primary School Children in Communities in Enugu, Nigeria
Authors: Nkeiruka Elsie Ezomike, Chinwe L. Onyekonwu, Anthony N. Ikefuna, Bede C. Ibe
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Superficial fungal infections (SFIs) are one of the common cutaneous infections that affect children worldwide. They may lead to school absenteeism or school drop-out and hence setback in the education of the child. Community-based studies in any locality are good reflections of the health conditions within that area. There is a dearth of information in the literature about SFI among primary school children in Enugu. This study aimed to determine the clinicomycological pattern of SFIs among primary school children in rural and urban communities in Enugu. This was a comparative descriptive cross-sectional study among primary school children in Awgu (rural) and Enugu North (urban) Local Government Areas (LGAs). Subjects' selection was made over 6 months using a multi-stage sampling method. Information such as age, sex, parental education, and occupation were collected using questionnaires. Socioeconomic classes of the children were determined using the classification proposed by Oyedeji et al. The samples were collected from subjects with SFIs. Potassium hydroxide tests were done on the samples. The samples that tested positive were cultured for SFI by inoculating onto Sabouraud's dextrose chloramphenicol actidione agar. The characteristics of the isolates were identified according to their morphological features using Mycology Online, Atlas 2000, and Mycology Review 2003. Equal numbers of children were recruited from the two LGAs. A total of 1662 pupils were studied. The mean ages of the study subjects were 9.03 ± 2.10years in rural and 10.46 ± 2.33years in urban communities. The male to female ratio was 1.6:1 in rural and 1:1.1 in urban communities. The personal hygiene of the children was significantly related to the presence of SFIs. The overall prevalence of SFIs among the study participants was 45%. In the rural, the prevalence was 29.6%, and in the urban prevalence was 60.4%. The types of SFIs were tinea capitis (the commonest), tinea corporis, pityriasis Versicolor, tinea unguium, and tinea manuum with prevalence rates lower in rural than urban communities. The clinical patterns were gray patch and black dot type of non-inflammatory tinea capitis, kerion, tinea corporis with trunk and limb distributions, and pityriasis Versicolor with face, trunk and limb distributions. Gray patch was the most frequent pattern of SFI seen in rural and urban communities. Black dot type was more frequent in rural than urban communities. SFIs were frequent among children aged 5 to 8years in rural and 9 to 12 years in urban communities. SFIs were commoner in males in the rural, whereas female dominance was observed in the urban. SFIs were more in children from low social class and those with poor hygiene. Trichophyton tonsurans and Trichophyton soudanese were the common mycological isolates in rural and urban communities, respectively. In conclusion, SFIs were less prevalent in rural than in urban communities. Trichophyton species were the most common fungal isolates in the communities. Health education of mothers and their children on SFI and good personal hygiene will reduce the incidence of SFIs.Keywords: clinicomycological pattern, communities, primary school children, superficial fungal infections
Procedia PDF Downloads 125114 Thermodynamics of Aqueous Solutions of Organic Molecule and Electrolyte: Use Cloud Point to Obtain Better Estimates of Thermodynamic Parameters
Authors: Jyoti Sahu, Vinay A. Juvekar
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Electrolytes are often used to bring about salting-in and salting-out of organic molecules and polymers (e.g. polyethylene glycols/proteins) from the aqueous solutions. For quantification of these phenomena, a thermodynamic model which can accurately predict activity coefficient of electrolyte as a function of temperature is needed. The thermodynamics models available in the literature contain a large number of empirical parameters. These parameters are estimated using lower/upper critical solution temperature of the solution in the electrolyte/organic molecule at different temperatures. Since the number of parameters is large, inaccuracy can bethe creep in during their estimation, which can affect the reliability of prediction beyond the range in which these parameters are estimated. Cloud point of solution is related to its free energy through temperature and composition derivative. Hence, the Cloud point measurement can be used for accurate estimation of the temperature and composition dependence of parameters in the model for free energy. Hence, if we use a two pronged procedure in which we first use cloud point of solution to estimate some of the parameters of the thermodynamic model and determine the rest using osmotic coefficient data, we gain on two counts. First, since the parameters, estimated in each of the two steps, are fewer, we achieve higher accuracy of estimation. The second and more important gain is that the resulting model parameters are more sensitive to temperature. This is crucial when we wish to use the model outside temperatures window within which the parameter estimation is sought. The focus of the present work is to prove this proposition. We have used electrolyte (NaCl/Na2CO3)-water-organic molecule (Iso-propanol/ethanol) as the model system. The model of Robinson-Stokes-Glukauf is modified by incorporating the temperature dependent Flory-Huggins interaction parameters. The Helmholtz free energy expression contains, in addition to electrostatic and translational entropic contributions, three Flory-Huggins pairwise interaction contributions viz., and (w-water, p-polymer, s-salt). These parameters depend both on temperature and concentrations. The concentration dependence is expressed in the form of a quadratic expression involving the volume fractions of the interacting species. The temperature dependence is expressed in the form .To obtain the temperature-dependent interaction parameters for organic molecule-water and electrolyte-water systems, Critical solution temperature of electrolyte -water-organic molecules is measured using cloud point measuring apparatus The temperature and composition dependent interaction parameters for electrolyte-water-organic molecule are estimated through measurement of cloud point of solution. The model is used to estimate critical solution temperature (CST) of electrolyte water-organic molecules solution. We have experimentally determined the critical solution temperature of different compositions of electrolyte-water-organic molecule solution and compared the results with the estimates based on our model. The two sets of values show good agreement. On the other hand when only osmotic coefficients are used for estimation of the free energy model, CST predicted using the resulting model show poor agreement with the experiments. Thus, the importance of the CST data in the estimation of parameters of the thermodynamic model is confirmed through this work.Keywords: concentrated electrolytes, Debye-Hückel theory, interaction parameters, Robinson-Stokes-Glueckauf model, Flory-Huggins model, critical solution temperature
Procedia PDF Downloads 391113 In Vitro Propagation of Vanilla Planifolia Using Nodal Explants and Varied Concentrations of Naphthaleneacetic acid (NAA) and 6-Benzylaminopurine (BAP).
Authors: Jessica Arthur, Duke Amegah, Kingsley Akenten Wiafe
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Background: Vanilla planifolia is the only edible fruit of the orchid family (Orchidaceae) among the over 35,000 Orchidaceae species found worldwide. In Ghana, Vanilla was discovered in the wild, but it is underutilized for commercial production, most likely due to a lack of knowledge on the best NAA and BAP combinations for in vitro propagation to promote successfully regenerated plant acclimatization. The growing interest and global demand for elite Vanilla planifolia plants and natural vanilla flavour emphasize the need for an effective industrial-scale micropropagation protocol. Tissue culture systems are increasingly used to grow disease-free plants and reliable in vitro methods can also produce plantlets with typically modest proliferation rates. This study sought to develop an efficient protocol for in vitro propagation of vanilla using nodal explants by testing different concentrations of NAA and BAP, for the proliferation of the entire plant. Methods: Nodal explants with dormant axillary buds were obtained from year-old laboratory-grown Vanilla planifolia plants. MS media was prepared with a nutrient stock solution (containing macronutrients, micronutrients, iron solution and vitamins) and semi-solidified using phytagel. It was supplemented with different concentrations of NAA and BAP to induce multiple shoots and roots (0.5mg/L BAP with NAA at 0, 0.5, 1, 1.5, 2.0mg/L and vice-versa). The explants were sterilized, cultured in labelled test tubes and incubated at 26°C ± 2°C with 16/8 hours light/dark cycle. Data on shoot and root growth, leaf number, node number, and survival percentage were collected over three consecutive two-week periods. The data were square root transformed and subjected to ANOVA and LSD at a 5% significance level using the R statistical package. Results: Shoots emerged at 8 days and roots at 12 days after inoculation with 94% survival rate. It was discovered that for the NAA treatments, MS media supplemented with 2.00 mg/l NAA resulted in the highest shoot length (10.45cm), maximum root number (1.51), maximum shoot number (1.47) and the highest number of leaves (1.29). MS medium containing 1.00 mg/l NAA produced the highest number of nodes (1.62) and root length (14.27cm). Also, a similar growth pattern for the BAP treatments was observed. MS medium supplemented with 1.50 mg/l BAP resulted in the highest shoot length (14.98 cm), the highest number of nodes (4.60), the highest number of leaves (1.75) and the maximum shoot number (1.57). MS medium containing 0.50 mg/l BAP and 1.0 mg/l BAP generated a maximum root number (1.44) and the highest root length (13.25cm), respectively. However, the best concentration combination for maximizing shoot and root was media containing 1.5mg/l BAP combined with 0.5mg/l NAA, and 1.0mg/l NAA combined with 0.5mg/l of BAP respectively. These concentrations were optimum for in vitro growth and production of Vanilla planifolia. Significance: This study presents a standardized protocol for labs to produce clean vanilla plantlets, enhancing cultivation in Ghana and beyond. It provides insights into Vanilla planifolia's growth patterns and hormone responses, aiding future research and cultivation.Keywords: Vanilla planifolia, In vitro propagation, plant hormones, MS media
Procedia PDF Downloads 67112 Circulating Public Perception on Agroforestry: Discourse Networks Analysis Using Social Media and Online News Media in Four Countries of the Sahel Region
Authors: Luisa Müting, Wisnu Harto Adiwijoyo
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Agroforestry systems transform the agricultural landscapes in the Sahel region of Africa, providing food and farming products consumed for subsistence or sold for income. In the incrementally dry climate of the Sahel region, the spreading of agroforestry practices is integral for policymaker efforts to counteract land degradation and provide soil restoration in the region. Several measures on agroforestry practices have been implemented in the region by governmental and non-governmental institutions in recent years. However, despite the efforts, past research shows that awareness of how policies and interventions are being consumed and perceived by the public remains low. Therefore, interpreting public policy dilemmas by analyzing the public perception regarding agroforestry concepts and practices is necessary. Public perceptions and discourses can be an essential driver or constraint for the adoption of agroforestry practices in the region. Thus, understanding the public discourse behavior of crucial stakeholders could assist policymakers in developing inclusive and contextual policies that are relevant to the context of agroforestry adoption in Sahel region. To answer how information about agroforestry spreads and is perceived by the public. As internet usage increased drastically over the past decade, reaching a share of 33 percent of the population being connected to the internet, this research is based on online conversation data. Social media data from Facebook are gathered daily between April 2021 and April 2022 in Djibouti, Senegal, Mali, and Nigeria based on their share of active internet users compared to other countries in the Sahel region. A systematic methodology was applied to the extracted social media using discourse network analysis (DNA). This study then clustered the data by the types of agroforestry practices, sentiments, and country. Additionally, this research extracted the text data from online news media during the same period to pinpoint events related to the topic of agroforestry. The preliminary result indicates that tree management, crops, and livestock integration, diversifying species and genetic resources, and focusing on interactions and productivity across the agricultural system; are the most notable keywords in agroforestry-related conversations within the four countries in the Sahel region. Additionally, approximately 84 percent of the discussions were still dominated by big actors, such as NGO or government actors. Furthermore, as a subject of communication within agroforestry discourse, the Great Green Wall initiative generates almost 60 percent positive sentiment within the captured social media data, effectively having a more significant outreach than general agroforestry topics. This study provides an understanding for scholars and policymakers with a springboard for further research or policy design on agroforestry in the four countries of the Sahel region with systematically uncaptured novel data from the internet.Keywords: sahel, djibouti, senegal, mali, nigeria, social networks analysis, public discourse analysis, sentiment analysis, content analysis, social media, online news, agroforestry, land restoration
Procedia PDF Downloads 102111 Phorbol 12-Myristate 13-Acetate (PMA)-Differentiated THP-1 Monocytes as a Validated Microglial-Like Model in Vitro
Authors: Amelia J. McFarland, Andrew K. Davey, Shailendra Anoopkumar-Dukie
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Microglia are the resident macrophage population of the central nervous system (CNS), contributing to both innate and adaptive immune response, and brain homeostasis. Activation of microglia occurs in response to a multitude of pathogenic stimuli in their microenvironment; this induces morphological and functional changes, resulting in a state of acute neuroinflammation which facilitates injury resolution. Adequate microglial function is essential for the health of the neuroparenchyma, with microglial dysfunction implicated in numerous CNS pathologies. Given the critical role that these macrophage-derived cells play in CNS homeostasis, there is a high demand for microglial models suitable for use in neuroscience research. The isolation of primary human microglia, however, is both difficult and costly, with microglial activation an unwanted but inevitable result of the extraction process. Consequently, there is a need for the development of alternative experimental models which exhibit morphological, biochemical and functional characteristics of human microglia without the difficulties associated with primary cell lines. In this study, our aim was to evaluate whether THP-1 human peripheral blood monocytes would display microglial-like qualities following an induced differentiation, and, therefore, be suitable for use as surrogate microglia. To achieve this aim, THP-1 human peripheral blood monocytes from acute monocytic leukaemia were differentiated with a range of phorbol 12-myristate 13-acetate (PMA) concentrations (50-200 nM) using two different protocols: a 5-day continuous PMA exposure or a 3-day continuous PMA exposure followed by a 5-day rest in normal media. In each protocol and at each PMA concentration, microglial-like cell morphology was assessed through crystal violet staining and the presence of CD-14 microglial / macrophage cell surface marker. Lipopolysaccharide (LPS) from Escherichia coli (055: B5) was then added at a range of concentrations from 0-10 mcg/mL to activate the PMA-differentiated THP-1 cells. Functional microglial-like behavior was evaluated by quantifying the release of prostaglandin (PG)-E2 and pro-inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor (TNF)-α using mediator-specific ELISAs. Furthermore, production of global reactive oxygen species (ROS) and nitric oxide (NO) were determined fluorometrically using dichlorodihydrofluorescein diacetate (DCFH-DA) and diaminofluorescein diacetate (DAF-2-DA) respectively. Following PMA-treatment, it was observed both differentiation protocols resulted in cells displaying distinct microglial morphology from 10 nM PMA. Activation of differentiated cells using LPS significantly augmented IL-1β, TNF-α and PGE2 release at all LPS concentrations under both differentiation protocols. Similarly, a significant increase in DCFH-DA and DAF-2-DA fluorescence was observed, indicative of increases in ROS and NO production. For all endpoints, the 5-day continuous PMA treatment protocol yielded significantly higher mediator levels than the 3-day treatment and 5-day rest protocol. Our data, therefore, suggests that the differentiation of THP-1 human monocyte cells with PMA yields a homogenous microglial-like population which, following stimulation with LPS, undergo activation to release a range of pro-inflammatory mediators associated with microglial activation. Thus, the use of PMA-differentiated THP-1 cells represents a suitable microglial model for in vitro research.Keywords: differentiation, lipopolysaccharide, microglia, monocyte, neuroscience, THP-1
Procedia PDF Downloads 388110 Bacterial Community Diversity in Soil under Two Tillage Systems
Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis
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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis
Procedia PDF Downloads 246109 Influence of the Use of Fruits Byproducts on the Lipid Profile of Hermetia illucens, Tenebrio molitor and Zophoba morio Larvae
Authors: Rebeca P Ramos-Bueno, Maria Jose Gonzalez-Fernandez, Rosa M. Moreno-Zamora, Antonia Barros Heras, Yolanda Serrano Alonso, Carolina Sanchez Barranco
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Insects are a new source of fatty acids (FA), so they are considered a sustainable and environmentally friendly alternative for both animal feed and the human diet, and furthermore, their harvesting/rearing require a low-tech and low capital investment. For that reason, lipids obtained by insect breeding open interesting possibilities with alimentary and industrial purposes, i.e., the production of biodiesel. Particularly, certain insect species, especially during the larval stage, contain high proportions of fat which is highly dependent on their feed and stage of development. Among them, Hermetia illucens larvae can be bred on food wastes to produce fat- and protein-rich raw materials for food by-product management. So, insects can act as excellent bioconverters of organic waste to nutrient-rich materials. In this regard, the aim of the study was to evaluate the effects of fruit byproducts on the FA compositions of Tenebrio molitor, Zophoba morio, and H. illucens larvae. Firstly, oil was extracted with the green solvent ethyl acetate, and FA methyl ester was obtained and analyzed by GC to show the FA profile. In addition, the triacylglycerol (TAG) profile was obtained by HPLC. Dehydrated watermelon, tomato, and papaya by-products, as well as wheat-based control feed, were assayed. High FA content was reached by Z. morio larvae fed with all fruits; however, no differences were shown in lipid profile with any change. It is worth highlighting that both Z. morio and H. illucens could be selected as the best candidates for biodiesel production due to their high content of saturated FA. On the other hand, T. molitor larvae showed a higher content of monounsaturated FA than control larvae, whereas the n-6 polyunsaturated FA content decreased in larvae fed with fruits. This result indicates that the improvement of the FA profile of Tenebrio can depend on both the type of feeding and the intended use. The lipid profile of H. illucens larvae fed with papaya and tomato showed a slight increase in the content of α-linoleic acid (ALA, 18:3n3). This FA is the precursor of docosahexaenoic acid (DHA, 22:6n3), which plays an important role as a component of structural lipids in cell membranes as well as in the synthesis of eicosanoids, protecting and resolving. Also, it was evaluated the TAG profile of Z. morio larvae due to their highest oil content. The results showed a high oleic acid (OA, 18:1n9) content, which displays modulatory effects in a wide range of physiological functions, having anti-inflammatory and anti-atherogenic properties. In conclusion, this study clearly shows that Z. morio and H. illucens larvae constitute an alternative source of OA- and ALA-rich oils, respectively, which can be devoted for food use, as well as for using in the food and pharmaceutical industries, with agronomic implications. Finally, although the profile of Z. morio was not improved with fruit feeding, this kind of feeding could be used due to its low environmental impact.Keywords: fatty acids, fruit byproducts, Hermetia illucens, Zophoba morio, Tenebrio molitor, insect rearing
Procedia PDF Downloads 147108 Inventory and Pollinating Role of Bees (Hymenoptera: apoidea) on Turnip (Brassica rapa L.) and Radish (Raphanus sativus L.) (Brassicaceae) in Constantine Area (Algeria)
Authors: Benachour Karima
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Pollination is a key factor in crop production and the presence of insect pollinators, mainly wild bees, is essential for improving yields. In this work, visiting apoids of two vegetable crops, the turnip (Brassica rapa L.) and the radish (Raphanus sativus L.) (Brassicaceae) were recorded during flowering times of 2003 and 2004 in Constantine area (36°22’N 06°37’E, 660 m). The observations were conducted in a plot of approximately 308 m2 of the Institute of Nutrition, Food and Food Technology (University of Mentouri Brothers). To estimate the density of bees (per 100 flowers or m2), 07 plots (01m2 for each one) are defined from the edge of the culture and in the first two rows. From flowering and every two days, foraging insects are recorded from 09 am until 17 pm (Gmt+1).The purpose of visit (collecting nectar, pollen or both) and pollinating efficiency (estimated by the number of flowers visited per minute and the number of positive visits) were noted for the most abundant bees on flowers. The action of pollinating insects is measured by comparing seed yields of 07 plots covered with tulle with 07 other accessible to pollinators. 04 families of Apoidea: Apidae, Halictidae, Andrenidae and Megachilidae were observed on the two plants. On turnip, the honeybee is the most common visitor (on average 214visites/ m2), it is followed by the Halictidae Lasioglossum mediterraneum whose visits are less intense (20 individuals/m2). Visits by Andrenidae, represented by several species such as Andrena lagopus, A.flavipes, A.agilissima and A.rhypara were episodic. The honeybee collected mainly nectar, its visits were all potentially fertilizing (contact with stigma) and more frequent (on average 14 flowers/min. L.mediterraneum visited only 05 flrs/min, it collected mostly the two products together and all its visits were also positive. On radish, the wild bee Ceratina cucurbitina recorded the highest number of visits (on average 06 individuals/100flo wers), the Halictidae represented mainly by L.mediterraneum, and L.malachurum, L.pauxillum were less abundant. C.cucurbitina visited on average 10 flowers /min and all its visits are positive. Visits of Halictidae were less frequent (05-06 flowers/min) and not all fertilizing. Seed yield of Brassica rapa (average number of pods /plant, seeds/ pods and average weight of 1000 seeds) was significantly higher in the presence of pollinators. Similarly, the pods of caged plants gave a percentage of aborted seeds (10.3%) significantly higher than that obtained on free plants (4.12%), the pods of caged plants also gave a percentage of malformed seeds (1.9%) significantly higher than that of the free plants (0.9%). For radish, the seed yield in the presence and absence of insects are almost similar. Only the percentage of malformed seeds (3.8%) obtained from the pods of caged plants was significantly higher in comparison with pods of free plants (1.9%). Following these results, it is clear that pollinators especially bees are essential for the production and improvement of crop yields and therefore it is necessary to protect this fauna increasingly threatened.Keywords: foraging behavior, honey bee, radish, seed yield, turnip, wild bee
Procedia PDF Downloads 214107 Microbial Analysis of Street Vended Ready-to-Eat Meat around Thohoyandou Area, Vhembe District, Limpopo Province, RSA
Authors: Tshimangadzo Jeanette Raedani, Edgar Musie, Afsatou Traore
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Background: Street-vended meats, including chicken, pork, and beef, are popular in urban areas worldwide due to their convenience and affordability. However, these meats often pose a significant risk of foodborne diseases. The high water activity, protein content, and nearly neutral pH of meat create conditions conducive to the growth of pathogenic bacteria. Street foods, particularly meats, are frequently linked to outbreaks of foodborne illnesses due to potential contamination from improper handling and preparation. This study aimed to assess the microbial quality and safety of street-vended ready-to-eat meat sold in the Thohoyandou area. Method: The study involved collecting 168 samples of street-vended meat, split evenly between chicken (n=84) and beef (n=84), from various vendors around Thohoyandou. The samples were randomly selected and transported in sterile conditions to the Department of Food Microbiology at the University of Venda for analysis. Each 10-gram sample was cultured in selective media: MSA for Staphylococcus aureus, EMB for E. coli O157, XLD agar for Salmonella, and Sorbitol McConkey for Shigella. After initial culturing, the presumptive colonies were sub-cultured for purification and identified through Gram staining and biochemical tests, including Catalase, API 20E, Klingler Iron Agar Test, and Vitek 2 system. Antibiotic susceptibility was tested using agents such as Ampicillin, Chloramphenicol, Penicillin, Neomycin, Tetracycline, Streptomycin, and Amoxicillin. Molecular characterization was performed to identify E. coli pathotypes using multiplex PCR. Results: Out of 168 samples tested, 32 (19%) were positive for Staphylococcus spp., with the highest prevalence found in cooked chicken meat. The most common staphylococcus species identified were S. xylosus (13.2%) and S. saprophyticus (10.5%). E. coli was present in 29 (19.3%) of the samples, with the highest prevalence in fried chicken. Antibiotic susceptibility testing showed that 100% of E. coli isolates were resistant to Ampicillin, Tetracycline, and Penicillin, but 100% were susceptible to Neomycin. Staphylococcus spp. isolates were also 100% resistant to Ampicillin and 100% susceptible to Neomycin. The study detected a range of virulence genes in E. coli, with prevalence rates from 13.33% to 86.67%. The identified pathotypes included EPEC, EHEC, ETEC, EAEC, and EIEC, with many isolates showing mixed pathotypes. Conclusion: The study highlighted that the microbial quality and safety of street-vended meats in Thohoyandou are inadequate, rendering them unsafe for consumption. The presence of pathogenic microorganisms in both beef and chicken samples indicates significant risks associated with poor personal hygiene and food preparation practices. This underscores the need for improved monitoring and stricter food safety measures to prevent foodborne diseases and ensure consumer safety.Keywords: meat, microbial analysis, street vendors, E. coli
Procedia PDF Downloads 27106 Altered Proteostasis Contributes to Skeletal Muscle Atrophy during Chronic Hypobaric Hypoxia: An Insight into Signaling Mechanisms
Authors: Akanksha Agrawal, Richa Rathor, Geetha Suryakumar
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Muscle represents about ¾ of the body mass, and a healthy muscular system is required for human performance. A healthy muscular system is dynamically balanced via the catabolic and anabolic process. High altitude associated hypoxia altered this redox balance via producing reactive oxygen and nitrogen species that ultimately modulates protein structure and function, hence, disrupts proteostasis or protein homeostasis. The mechanism by which proteostasis is clinched includes regulated protein translation, protein folding, and protein degradation machinery. Perturbation in any of these mechanisms could increase proteome imbalance in the cellular processes. Altered proteostasis in skeletal muscle is likely to be responsible for contributing muscular atrophy in response to hypoxia. Therefore, we planned to elucidate the mechanism involving altered proteostasis leading to skeletal muscle atrophy under chronic hypobaric hypoxia. Material and Methods-Male Sprague Dawley rats weighing about 200-220 were divided into five groups - Control (Normoxic animals), 1d, 3d, 7d and 14d hypobaric hypoxia exposed animals. The animals were exposed to simulated hypoxia equivalent to 282 torr pressure (equivalent to an altitude of 7620m, 8% oxygen) at 25°C. On completion of chronic hypobaric hypoxia (CHH) exposure, rats were sacrificed, muscle was excised and biochemical, histopathological and protein synthesis signaling were studied. Results-A number of changes were observed with the CHH exposure time period. ROS was increased significantly on 07 and 14 days which were attributed to protein oxidation via damaging muscle protein structure by oxidation of amino acids moiety. The oxidative damage to the protein further enhanced the various protein degradation pathways. Calcium activated cysteine proteases and other intracellular proteases participate in protein turnover in muscles. Therefore, we analysed calpain and 20S proteosome activity which were noticeably increased at CHH exposure as compared to control group representing enhanced muscle protein catabolism. Since inflammatory markers (myokines) affect protein synthesis and triggers degradation machinery. So, we determined inflammatory pathway regulated under hypoxic environment. Other striking finding of the study was upregulation of Akt/PKB translational machinery that was increased on CHH exposure. Akt, p-Akt, p70 S6kinase, and GSK- 3β expression were upregulated till 7d of CHH exposure. Apoptosis related markers, caspase-3, caspase-9 and annexin V was also increased on CHH exposure. Conclusion: The present study provides evidence of disrupted proteostasis under chronic hypobaric hypoxia. A profound loss of muscle mass is accompanied by the muscle damage leading to apoptosis and cell death under CHH. These cellular stress response pathways may play a pivotal role in hypobaric hypoxia induced skeletal muscle atrophy. Further research in these signaling pathways will lead to development of therapeutic interventions for amelioration of hypoxia induced muscle atrophy.Keywords: Akt/PKB translational machinery, chronic hypobaric hypoxia, muscle atrophy, protein degradation
Procedia PDF Downloads 270105 Numerical Model of Crude Glycerol Autothermal Reforming to Hydrogen-Rich Syngas
Authors: A. Odoom, A. Salama, H. Ibrahim
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Hydrogen is a clean source of energy for power production and transportation. The main source of hydrogen in this research is biodiesel. Glycerol also called glycerine is a by-product of biodiesel production by transesterification of vegetable oils and methanol. This is a reliable and environmentally-friendly source of hydrogen production than fossil fuels. A typical composition of crude glycerol comprises of glycerol, water, organic and inorganic salts, soap, methanol and small amounts of glycerides. Crude glycerol has limited industrial application due to its low purity thus, the usage of crude glycerol can significantly enhance the sustainability and production of biodiesel. Reforming techniques is an approach for hydrogen production mainly Steam Reforming (SR), Autothermal Reforming (ATR) and Partial Oxidation Reforming (POR). SR produces high hydrogen conversions and yield but is highly endothermic whereas POR is exothermic. On the downside, PO yields lower hydrogen as well as large amount of side reactions. ATR which is a fusion of partial oxidation reforming and steam reforming is thermally neutral because net reactor heat duty is zero. It has relatively high hydrogen yield, selectivity as well as limits coke formation. The complex chemical processes that take place during the production phases makes it relatively difficult to construct a reliable and robust numerical model. Numerical model is a tool to mimic reality and provide insight into the influence of the parameters. In this work, we introduce a finite volume numerical study for an 'in-house' lab-scale experiment of ATR. Previous numerical studies on this process have considered either using Comsol or nodal finite difference analysis. Since Comsol is a commercial package which is not readily available everywhere and lab-scale experiment can be considered well mixed in the radial direction. One spatial dimension suffices to capture the essential feature of ATR, in this work, we consider developing our own numerical approach using MATLAB. A continuum fixed bed reactor is modelled using MATLAB with both pseudo homogeneous and heterogeneous models. The drawback of nodal finite difference formulation is that it is not locally conservative which means that materials and momenta can be generated inside the domain as an artifact of the discretization. Control volume, on the other hand, is locally conservative and suites very well problems where materials are generated and consumed inside the domain. In this work, species mass balance, Darcy’s equation and energy equations are solved using operator splitting technique. Therefore, diffusion-like terms are discretized implicitly while advection-like terms are discretized explicitly. An upwind scheme is adapted for the advection term to ensure accuracy and positivity. Comparisons with the experimental data show very good agreements which build confidence in our modeling approach. The models obtained were validated and optimized for better results.Keywords: autothermal reforming, crude glycerol, hydrogen, numerical model
Procedia PDF Downloads 140104 Vascular Targeted Photodynamic Therapy Monitored by Real-Time Laser Speckle Imaging
Authors: Ruth Goldschmidt, Vyacheslav Kalchenko, Lilah Agemy, Rachel Elmoalem, Avigdor Scherz
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Vascular Targeted Photodynamic therapy (VTP) is a new modality for selective cancer treatment that leads to the complete tumor ablation. A photosensitizer, a bacteriochlorophyll derivative in our case, is first administered to the patient and followed by the illumination of the tumor area, by a near-IR laser for its photoactivation. The photoactivated drug releases reactive oxygen species (ROS) in the circulation, which reacts with blood cells and the endothelium leading to the occlusion of the blood vasculature. If the blood vessels are only partially closed, the tumor may recover, and cancer cells could survive. On the other hand, excessive treatment may lead to toxicity of healthy tissues nearby. Simultaneous VTP monitoring and image processing independent of the photoexcitation laser has not yet been reported, to our knowledge. Here we present a method for blood flow monitoring, using a real-time laser speckle imaging (RTLSI) in the tumor during VTP. We have synthesized over the years a library of bacteriochlorophyll derivatives, among them WST11 and STL-6014. Both are water soluble derivatives that are retained in the blood vasculature through their partial binding to HSA. WST11 has been approved in Mexico for VTP treatment of prostate cancer at a certain drug dose, and time/intensity of illumination. Application to other bacteriochlorophyll derivatives or other cancers may require different treatment parameters (such as light/drug administration). VTP parameters for STL-6014 are still under study. This new derivative mainly differs from WST11 by its lack of the central Palladium, and its conjugation to an Arg-Gly-Asp (RGD) sequence. RGD is a tumor-specific ligand that is used for targeting the necrotic tumor domains through its affinity to αVβ3 integrin receptors. This enables the study of cell-targeted VTP. We developed a special RTLSI module, based on Labview software environment for data processing. The new module enables to acquire raw laser speckle images and calculate the values of the laser temporal statistics of time-integrated speckles in real time, without additional off-line processing. Using RTLSI, we could monitor the tumor’s blood flow following VTP in a CT26 colon carcinoma ear model. VTP with WST11 induced an immediate slow down of the blood flow within the tumor and a complete final flow arrest, after some sporadic reperfusions. If the irradiation continued further, the blood flow stopped also in the blood vessels of the surrounding healthy tissue. This emphasizes the significance of light dose control. Using our RTLSI system, we could prevent any additional healthy tissue damage by controlling the illumination time and restrict blood flow arrest within the tumor only. In addition, we found that VTP with STL-6014 was the most effective when the photoactivation was conducted 4h post-injection, in terms of tumor ablation success in-vivo and blood vessel flow arrest. In conclusion, RTSLI application should allow to optimize VTP efficacy vs. toxicity in both the preclinical and clinical arenas.Keywords: blood vessel occlusion, cancer treatment, photodynamic therapy, real time imaging
Procedia PDF Downloads 223103 Aerobic Biodegradation of a Chlorinated Hydrocarbon by Bacillus Cereus 2479
Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay
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Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.Keywords: cloning, Bacillus cereus, in silico analysis, TCE
Procedia PDF Downloads 398102 South-Mediterranean Oaks Forests Management in Changing Climate Case of the National Park of Tlemcen-Algeria
Authors: K. Bencherif, M. Bellifa
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The expected climatic changes in North Africa are the increase of both intensity and frequencies of the summer droughts and a reduction in water availability during growing season. The exiting coppices and forest formations in the national park of Tlemcen are dominated by holm oak, zen oak and cork oak. These opened-fragmented structures don’t seem enough strong so to hope durable protection against climate change. According to the observed climatic tendency, the objective is to analyze the climatic context and its evolution taking into account the eventual behaving of the oak species during the next 20-30 years on one side and the landscaped context in relation with the most adequate sylvicultural models to choose and especially in relation with human activities on another side. The study methodology is based on Climatic synthesis and Floristic and spatial analysis. Meteorological data of the decade 1989-2009 are used to characterize the current climate. An another approach, based on dendrochronological analysis of a 120 years sample Aleppo pine stem growing in the park, is used so to analyze the climate evolution during one century. Results on the climate evolution during the 50 years obtained through climatic predictive models are exploited so to predict the climate tendency in the park. Spatially, in each forest unit of the Park, stratified sampling is achieved so to reduce the degree of heterogeneity and to easily delineate different stands using the GPS. Results from precedent study are used to analyze the anthropogenic factor considering the forecasts for the period 2025-2100, the number of warm days with a temperature over 25°C would increase from 30 to 70. The monthly mean temperatures of the maxima’s (M) and the minima’s (m) would pass respectively from 30.5°C to 33°C and from 2.3°C to 4.8°C. With an average drop of 25%, precipitations will be reduced to 411.37 mm. These new data highlight the importance of the risk fire and the water stress witch would affect the vegetation and the regeneration process. Spatial analysis highlights the forest and the agricultural dimensions of the park compared to the urban habitat and bare soils. Maps show both fragmentation state and forest surface regression (50% of total surface). At the level of the park, fires affected already all types of covers creating low structures with various densities. On the silvi cultural plan, Zen oak form in some places pure stands and this invasion must be considered as a natural tendency where Zen oak becomes the structuring specie. Climate-related changes have nothing to do with the real impact that South-Mediterranean forests are undergoing because human constraints they support. Nevertheless, hardwoods stand of oak in the national park of Tlemcen will face up to unexpected climate changes such as changing rainfall regime associated with a lengthening of the period of water stress, to heavy rainfall and/or to sudden cold snaps. Faced with these new conditions, management based on mixed uneven aged high forest method promoting the more dynamic specie could be an appropriate measure.Keywords: global warming, mediterranean forest, oak shrub-lands, Tlemcen
Procedia PDF Downloads 389101 Traditional Practices of Conserving Biodiversity: A Case Study around Jim Corbett National Park, Uttarakhand, India
Authors: Rana Parween, Rob Marchant
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With the continued loss of global biodiversity despite the application of modern conservation techniques, it has become crucial to investigate non-conventional methods. Accelerated destruction of ecosystems due to altered land use, climate change, cultural and social change, necessitates the exploration of society-biodiversity attitudes and links. While the loss of species and their extinction is a well-known and well-documented process that attracts much-needed attention from researchers, academics, government and non-governmental organizations, the loss of traditional ecological knowledge and practices is more insidious and goes unnoticed. The growing availability of 'indirect experiences' such as the internet and media are leading to a disaffection towards nature and the 'Extinction of Experience'. Exacerbated by the lack of documentation of traditional practices and skills, there is the possibility for the 'extinction' of traditional practices and skills before they are fully recognized and captured. India, as a mega-biodiverse country, is also known for its historical conservation strategies entwined in traditional beliefs. Indigenous communities hold skillsets, knowledge, and traditions that have accumulated over multiple generations and may play an important role in conserving biodiversity today. This study explores the differences in knowledge and attitudes towards conserving biodiversity, of three different stakeholder groups living around Jim Corbett National Park, based on their age, traditions, and association with the protected area. A triangulation designed multi-strategy investigation collected qualitative and quantitative data through a questionnaire survey of village elders, the general public, and forest officers. Following an inductive approach to analyzing qualitative data, the thematic content analysis was followed. All coding and analysis were completed using NVivo 11. Although the village elders and some general public had vast amounts of traditional knowledge, most of it was related to animal husbandry and the medicinal value of plants. Village elders were unfamiliar with the concept of the term ‘biodiversity’ albeit their way of life and attitudes ensured that they care for the ecosystem without having the scientific basis underpinning biodiversity conservation. Inherently, village elders were keen to conserve nature; the superimposition of governmental policies without any tangible benefit or consultation was seen as detrimental. Alienating villagers and consequently the village elders who are the reservoirs of traditional knowledge would not only be damaging to the social network of the area but would also disdain years of tried and tested techniques held by the elders. Forest officers advocated for biodiversity and conservation education for women and children. Women, across all groups, when questioned about nature conservation, showed more interest in learning and participation. Biodiversity not only has an ethical and cultural value, but also plays a role in ecosystem function and, thus, provides ecosystem services and supports livelihoods. Therefore, underpinning and using traditional knowledge and incorporating them into programs of biodiversity conservation should be explored with a sense of urgency.Keywords: biological diversity, mega-biodiverse countries, traditional ecological knowledge, society-biodiversity links
Procedia PDF Downloads 105100 Arthropods Diversity of the Late Carboniferous Souss Basin, Morocco: Paleoecology and Taphonomy
Authors: Abouchouaib Belahmira, Joerg W. Schneider, Hafid Saber
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Continental sediments of the uppermost Carboniferous (late Pennsylvanian) El Menizla and Oued Issene formations of the Souss basin, Southwestern High Atlas Mountains, Morocco have yielded abundant well-preserved arthropods. The latter comprise freshwater and terrestrial elements, were found associated with plants, freshwater jellyfish and pelecypods. Arthropods are ubiquitous and typically restricted to the dominated lacustrine black shale taphofacies. The lithofacies interpretation and its correlation with the taphofacies led to the determination of the original depositional environment that was reconstructed as a fluvial-dominated with braided wide channel system and floodplain lakes to peat local backswamps sub-environments. The late Carboniferous fossiliferous strata have been correlated biostratigraphically with many other Pennsylvanian (Kasimovian/Gzhelian) deposits of North America and Europe on the basis of entomological studies. The faunal elements of the lentic biocoenosis of the Souss basin are depauperate, with the vagile forms slightly diverse than sessile ones. The prevailing groups are small shelly fauna, other habitat guild such as apterygotan Monura insect dasyleptids. The fossils recorded from the Souss basin includes crustaceans, of various sizes (µm- to mm) and morphologies, preservation state ranging from poorly preserved to rarely well-preserved specimens. Their remains sporadically found clustered and preserved as internal or external shell molds or steinkerns often disarticulated specimens. Ostracods as more likely Carbonita, their shells are preserved three-dimensionally. The clam shrimps conchostracans record of the Souss basin are often determined as pseudestherids and the Spinicaudatan leaiids. The moldic preservation is somewhat similar to pelecypods, they are known from internal casts or impressions. Monura insects are characterized by their low diversity, thus, only two species are known Dasyleptus lucasi Brongniart and Dasyleptus noli Rasnitsyn. The terrestrial component consists of pterygotan insects. They are diverse, significantly more frequent throughout the Souss basin fossil localities, numerically dominated by the members of Blattodea (cockroaches). The fossil record includes Blattodea, Protorthoptera, Diaphanopterodea, Ephemeroptera (mayfly) , Calneurodea, Grylloblattodea, Miomoptera and Palaeodictyoptera. Additionally, the composition of the preserved insect is mostly represented by completely isolated forewings, rare membranous hindwings, parts of the body or exceptionally preserved specimens, which may reflect a wide spectrum of taphonomic pathways. The steady increase in taxonomic diversity of fossil sites in the Souss basin, together with the taphonomic interpretation of arthropods assemblages, have contributed to provide a novel insight into the complex terrestrial ecosystem that thrived in this paleotropical key region during the late Pennsylvanian and additionally to understand climate-driven paleobiogeography and paleoecology of late Paleozoic non-marine arthropods.Keywords: Souss, carboniferous, arthropods, taphonomy, paleoecology.
Procedia PDF Downloads 3399 Changes of Mitochondrial Potential in the Midgut Epithelium of Lithobius forficatus (Myriapoda, Chilopoda) Exposed to Cadmium Concentrated in Soil
Authors: Magdalena Rost-Roszkowska, Izabela Poprawa, Alina Chachulska-Zymelka, Lukasz Chajec, Grazyna Wilczek, Piotr Wilczek, Malgorzata Lesniewska
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Lithobius forficatus, commonly known as the brown centipede, is a widespread European species, which lives in the upper layers of soil, under stones, litter, rocks, and leaves. As the soil organism, it is exposed to numerous stressors such as xenobiotics, including heavy metals, temperature, starvation, pathogens, etc. Heavy metals are treated as the environmental pollutants of the soil because of their toxic effects on plants, animals and human being. One of the heavy metals which is xenobiotic and can be taken up by plants or animals from the soil is cadmium. The digestive system of centipedes is composed of three distinct regions: fore-, mid- and hindgut. The salivary glands of centipedes are the organs which belong to the anterior region of the digestive system and take part in the synthesis, accumulation, and secretion of many substances. The middle region having contact with the food masses is treated as one of the barriers which protect the organism against any stressors which originate from the external environment, e.g., toxic metals. As the material for our studies, we chose two organs of the digestive system in brown centipede, the organs which take part in homeostasis maintenance: the salivary glands and the midgut. The main purpose of the project was to investigate the relationship between the percentage of depolarized mitochondria, mitophagy and ATP level in cells of mentioned above organs. The animals were divided into experimental groups: K – the control group, the animals cultured in a laboratory conditions in a horticultural soil and fed with Acheta domesticus larvae; Cd1 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2, fed with A. domesticus larvae maintained in tap water, 12 days – short-term exposure; Cd2 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2, fed with A. domesticus larvae maintained in tap water, 45 days – long-term exposure. The studies were conducted using transmission electron microscopy (TEM), flow cytometry and confocal microscopy. Quantitative analysis revealed that regardless of the organ, a progressive increase in the percentage of cells with depolarized mitochondria was registered, but only in the salivary glands. These were statistically significant changes from the control. In both organs, there were no differences in the level of the analyzed parameter depending on the duration of exposure of individuals to cadmium. Changes in the ultrastructure of mitochondria have been observed. With the extension of the body's exposure time to metal, an increase in the ADP/ATP index was recorded. However, changes statistically significant to the control were demonstrated in the intestine and salivary glands. The size of this intestinal index and salivary glands in the Cd2 group was about thirty and twenty times higher, respectively than in control. Acknowledgment: The study has been financed by the National Science Centre, Poland, grant no 2017/25/B/NZ4/00420.Keywords: cadmium, digestive system, ultrastructure, centipede
Procedia PDF Downloads 13698 Biomimicked Nano-Structured Coating Elaboration by Soft Chemistry Route for Self-Cleaning and Antibacterial Uses
Authors: Elodie Niemiec, Philippe Champagne, Jean-Francois Blach, Philippe Moreau, Anthony Thuault, Arnaud Tricoteaux
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Hygiene of equipment in contact with users is an important issue in the railroad industry. The numerous cleanings to eliminate bacteria and dirt cost a lot. Besides, mechanical solicitations on contact parts are observed daily. It should be interesting to elaborate on a self-cleaning and antibacterial coating with sufficient adhesion and good resistance against mechanical and chemical solicitations. Thus, a Hauts-de-France and Maubeuge Val-de-Sambre conurbation authority co-financed Ph.D. thesis has been set up since October 2017 based on anterior studies carried by the Laboratory of Ceramic Materials and Processing. To accomplish this task, a soft chemical route has been implemented to bring a lotus effect on metallic substrates. It involves nanometric liquid zinc oxide synthesis under 100°C. The originality here consists in a variation of surface texturing by modification of the synthesis time of the species in solution. This helps to adjust wettability. Nanostructured zinc oxide has been chosen because of the inherent photocatalytic effect, which can activate organic substance degradation. Two methods of heating have been compared: conventional and microwave assistance. Tested subtracts are made of stainless steel to conform to transport uses. Substrate preparation was the first step of this protocol: a meticulous cleaning of the samples is applied. The main goal of the elaboration protocol is to fix enough zinc-based seeds to make them grow during the next step as desired (nanorod shaped). To improve this adhesion, a silica gel has been formulated and optimized to ensure chemical bonding between substrate and zinc seeds. The last step consists of deposing a wide carbonated organosilane to improve the superhydrophobic property of the coating. The quasi-proportionality between the reaction time and the nanorod length will be demonstrated. Water Contact (superior to 150°) and Roll-off Angle at different steps of the process will be presented. The antibacterial effect has been proved with Escherichia Coli, Staphylococcus Aureus, and Bacillus Subtilis. The mortality rate is found to be four times superior to a non-treated substrate. Photocatalytic experiences were carried out from different dyed solutions in contact with treated samples under UV irradiation. Spectroscopic measurements allow to determinate times of degradation according to the zinc quantity available on the surface. The final coating obtained is, therefore, not a monolayer but rather a set of amorphous/crystalline/amorphous layers that have been characterized by spectroscopic ellipsometry. We will show that the thickness of the nanostructured oxide layer depends essentially on the synthesis time set in the hydrothermal growth step. A green, easy-to-process and control coating with self-cleaning and antibacterial properties has been synthesized with a satisfying surface structuration.Keywords: antibacterial, biomimetism, soft-chemistry, zinc oxide
Procedia PDF Downloads 14297 Response of Subfossile Diatoms, Cladocera, and Chironomidae in Sediments of Small Ponds to Changes in Wastewater Discharges from a Zn–Pb Mine
Authors: Ewa Szarek-Gwiazda, Agata Z. Wojtal, Agnieszka Pociecha, Andrzej Kownacki, Dariusz Ciszewski
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Mining of metal ores is one of the largest sources of heavy metals, which deteriorate aquatic systems. The response of organisms to environmental changes can be well recorded in sediments of the affected water bodies and may be reconstructed based on analyses of organisms' remains. The present study aimed at the response of diatoms (Bacillariophyta), Cladocera, and Chironomidae communities to the impact of Zn-Pb mine water discharge recorded in sediment cores of small subsidence ponds on the Chechło River floodplain (Silesia–Krakow Region, southern Poland). We hypothesize various responses of the above groups to high metal concentrations (Cd, Pb, Zn, and Cu). The investigated ponds were formed either during the peak of the ore exploitation (DOWN) or after mining cessation (UP). Currently, the concentrations of dissolved metals (in µg g⁻¹) in water reached up to 0.53 for Cd, 7.3 for Pb, and up to 47.1 for Zn. All the sediment cores from subsidence ponds were heavily polluted with Cd 6.7–612 μg g⁻¹, Pb 0.1–10.2 mg g⁻¹, and Zn 0.5–23.1 mg g⁻¹. Core sediments varied also in respect to pH 5.8-7.1 and concentrations of organic matter (5.7-39.8%). The impact of high metal concentrations was expressed by the occurrence of metal-tolerant taxa like diatoms – Nitzschia amphibia, Sellaphora nigri, and Surirella brebisonii var. kuetzingii; Cladocera – Chydorus sphaericus (dominated in cores from all ponds), and Chironomidae – Chironomus and Cricotopus especially in the DOWN ponds. Statistical analysis exhibited a negative impact of metals on some taxa of diatoms and Cladocera but only on Polypedilum sp. from Chironomidae. The abundance of such diatoms like Gomphonema utae, Staurosirella pinnata, Eunotia bilunaris, and Cladocera like Alona, Chydorus, Graptoleberis, and Pleuroxus decreased with increasing Pb concentration. However, the occurrence or dominance of more sensitive species of diatoms and Cladocera indicates their adaptation to higher metal loads, which was facilitated by neutral pH and slightly alkaline waters. Diatom assemblages were generally resistant to Zn, Pb, Cu, and Cd pollution, as indicated by their large similarity to populations from non-contaminated waters. Comparison with reference objects clearly indicates the dominance of Achnanthidium minutissimum, Staurosira venter, and Fragilaria gracilis in very diverse assemblages of unpolluted waters. The distribution of the Cladocera and Chironomidae taxa depended on the habitat type. The DOWN ponds with stagnant water and overgrown with macrophytes were more suitable for cladocerans (14 taxa, higher diversity) than the UP ponds with river water flowing through their centre and with a small share of macrophytes (8 taxa). The Chironominae, mainly Chironomus and Microspectra, were abundant in cores from the UP ponds with muddy bottoms. Inversely, the density of Orthocladiinae, especially genus Cricotopus, was related to the organic matter content and dominated in cores from the DOWN ponds. The presence of diatoms like Nitzschia amphibia, Sellaphora nigri, and Surirella brebisonii var. kuetzingii, cladocerans: Bosmina longirostris, Chydorus sphaericus, Alona affinis, and A. rectangularis as well as Chironomidae Chironomus sp. (UP ponds) and Psecrotanypus varius (DOWN ponds) indicate the influence of the water trophy on their distribution.Keywords: Chironomidae, Cladocera, diatoms, metals, Zn-Pb mine, sediment cores, subsidence ponds
Procedia PDF Downloads 7796 Co-Culture with Murine Stromal Cells Enhances the In-vitro Expansion of Hematopoietic Stem Cells in Response to Low Concentrations of Trans-Resveratrol
Authors: Mariyah Poonawala, Selvan Ravindran, Anuradha Vaidya
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Despite much progress in understanding the regulatory factors and cytokines that support the maturation of the various cell lineages of the hematopoietic system, factors that govern the self-renewal and proliferation of hematopoietic stem cells (HSCs) is still a grey area of research. Hematopoietic stem cell transplantation (HSCT) has evolved over the years and gained tremendous importance in the treatment of both malignant and non-malignant diseases. However, factors such as graft rejection and multiple organ failure have challenged HSCT from time to time, underscoring the urgent need for development of milder processes for successful hematopoietic transplantation. An emerging concept in the field of stem cell biology states that the interactions between the bone-marrow micro-environment and the hematopoietic stem and progenitor cells is essential for regulation, maintenance, commitment and proliferation of stem cells. Understanding the role of mesenchymal stromal cells in modulating the functionality of HSCs is, therefore, an important area of research. Trans-resveratrol has been extensively studied for its various properties to combat and prevent cancer, diabetes and cardiovascular diseases etc. The aim of the present study was to understand the effect of trans-resveratrol on HSCs using single and co-culture systems. We have used KG1a cells since it is a well accepted hematopoietic stem cell model system. Our preliminary experiments showed that low concentrations of trans-resveratrol stimulated the HSCs to undergo proliferation whereas high concentrations of trans-resveratrol did not stimulate the cells to proliferate. We used a murine fibroblast cell line, M210B4, as a stromal feeder layer. On culturing the KG1a cells with M210B4 cells, we observed that the stimulatory as well as inhibitory effects of trans-resveratrol at low and high concentrations respectively, were enhanced. Our further experiments showed that low concentration of trans-resveratrol reduced the generation of reactive oxygen species (ROS) and nitric oxide (NO) whereas high concentrations increased the oxidative stress in KG1a cells. We speculated that perhaps the oxidative stress was imposing inhibitory effects at high concentration and the same was confirmed by performing an apoptotic assay. Furthermore, cell cycle analysis and growth kinetic experiments provided evidence that low concentration of trans-resveratrol reduced the doubling time of the cells. Our hypothesis is that perhaps at low concentration of trans-resveratrol the cells get pushed into the G0/G1 phase and re-enter the cell cycle resulting in their proliferation, whereas at high concentration the cells are perhaps arrested at G2/M phase or at cytokinesis and therefore undergo apoptosis. Liquid Chromatography-Quantitative-Time of Flight–Mass Spectroscopy (LC-Q-TOF MS) analyses indicated the presence of trans-resveratrol and its metabolite(s) in the supernatant of the co-cultured cells incubated with high concentration of trans-resveratrol. We conjecture that perhaps the metabolites of trans-resveratrol are responsible for the apoptosis observed at the high concentration. Our findings may shed light on the unsolved problems in the in vitro expansion of stem cells and may have implications in the ex vivo manipulation of HSCs for therapeutic purposes.Keywords: co-culture system, hematopoietic micro-environment, KG1a cell line, M210B4 cell line, trans-resveratrol
Procedia PDF Downloads 25895 Unravelling Glyphosates Disruptive Effects on the Photochemical Efficiency of Amaranthus cruentus
Authors: Jacques M. Berner, Lehlogonolo Maloma
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Context: Glyphosate, a widely used herbicide, has raised concerns about its impact on various crops. Amaranthus cruentus, an important grain crop species, is particularly susceptible to glyphosate. Understanding the specific disruptions caused by glyphosate on the photosynthetic process in Amaranthus cruentus is crucial for assessing its effects on crop productivity and ecological sustainability. Research Aim: This study aimed to investigate the dose-dependent impact of glyphosate on the photochemical efficiency of Amaranthus cruentus using the OJIP transient analysis. The goal was to assess the specific disruptions caused by glyphosate on key parameters of photosystem II. Methodology: The experiment was conducted in a controlled greenhouse environment. Amaranthus cruentus plants were exposed to different concentrations of glyphosate, including half, recommended, and double the recommended application rates. The photochemical efficiency of the plants was evaluated using non-invasive chlorophyll a fluorescence measurements and subsequent analysis of OJIP transients. Measurements were taken on 1-hour dark-adapted leaves using a Hansatech Handy PEA+ chlorophyll fluorimeter. Findings: The study's results demonstrated a significant reduction in the photochemical efficiency of Amaranthus cruentus following glyphosate treatment. The OJIP transients showed distinct alterations in the glyphosate-treated plants compared to the control group. These changes included a decrease in maximal fluorescence (FP) and a delay in the rise of the fluorescence signal, indicating impairment in the energy conversion process within the photosystem II. Glyphosate exposure also led to a substantial decrease in the maximum quantum yield efficiency of photosystem II (FV/FM) and the total performance index (PItotal), which reflects the overall photochemical efficiency of photosystem II. These reductions in photochemical efficiency were observed even at half the recommended dose of glyphosate. Theoretical Importance: The study provides valuable insights into the specific disruptions caused by glyphosate on the photochemical efficiency of Amaranthus cruentus. Data Collection and Analysis Procedures: Data collection involved non-invasive chlorophyll a fluorescence measurements using a chlorophyll fluorimeter on dark-adapted leaves. The OJIP transients were then analyzed to assess specific disruptions in key parameters of photosystem II. Statistical analysis was conducted to determine the significance of the differences observed between glyphosate-treated plants and the control group. Question Addressed: The study aimed to address the question of how glyphosate exposure affects the photochemical efficiency of Amaranthus cruentus, specifically examining disruptions in the photosynthetic electron transport chain and overall photochemical efficiency. Conclusion: The study demonstrates that glyphosate severely impairs the photochemical efficiency of Amaranthus cruentus, as indicated by the alterations in OJIP transients. Even at half the recommended dose, glyphosate caused significant reductions in photochemical efficiency. These findings highlight the detrimental effects of glyphosate on crop productivity and emphasize the need for further research to evaluate its long-term consequences and ecological implications in agriculture. The authors gratefully acknowledge the support from North-West University for making this research possible.Keywords: glyphosate, amaranthus cruentus, ojip transient analysis, pitotal, photochemical efficiency, chlorophyll fluorescence, weeds
Procedia PDF Downloads 9194 Multi-Criteria Geographic Information System Analysis of the Costs and Environmental Impacts of Improved Overland Tourist Access to Kaieteur National Park, Guyana
Authors: Mark R. Leipnik, Dahlia Durga, Linda Johnson-Bhola
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Kaieteur is the most iconic National Park in the rainforest-clad nation of Guyana in South America. However, the magnificent 226-meter-high waterfall at its center is virtually inaccessible by surface transportation, and the occasional charter flights to the small airstrip in the park are too expensive for many tourists and residents. Thus, the largest waterfall in all of Amazonia, where the Potaro River plunges over a single free drop twice as high as Victoria Falls, remains preserved in splendid isolation inside a 57,000-hectare National Park established by the British in 1929, in the deepest recesses of a remote jungle canyon. Kaieteur Falls are largely unseen firsthand, but images of the falls are depicted on the Guyanese twenty dollar note, in every Guyanese tourist promotion, and on many items in the national capital of Georgetown. Georgetown is only 223-241 kilometers away from the falls. The lack of a single mileage figure demonstrates there is no single overland route. Any journey, except by air, involves changes of vehicles, a ferry ride, and a boat ride up a jungle river. It also entails hiking for many hours to view the falls. Surface access from Georgetown (or any city) is thus a 3-5 day-long adventure; even in the dry season, during the two wet seasons, travel is a particularly sticky proposition. This journey was made overland by the paper's co-author Dahlia Durga. This paper focuses on potential ways to improve overland tourist access to Kaieteur National Park from Georgetown. This is primarily a GIS-based analysis, using multiple criteria to determine the least cost means of creating all-weather road access to the area near the base of the falls while minimizing distance and elevation changes. Critically, it also involves minimizing the number of new bridges required to be built while utilizing the one existing ferry crossings of a major river. Cost estimates are based on data from road and bridge construction engineers operating currently in the interior of Guyana. The paper contains original maps generated with ArcGIS of the potential routes for such an overland connection, including the one deemed optimal. Other factors, such as the impact on endangered species habitats and Indigenous populations, are considered. This proposed infrastructure development is taking place at a time when Guyana is undergoing the largest boom in its history due to revenues from offshore oil and gas development. Thus, better access to the most important tourist attraction in the country is likely to happen eventually in some manner. But the questions of the most environmentally sustainable and least costly alternatives for such access remain. This paper addresses those questions and others related to access to this magnificent natural treasure and the tradeoffs such access will have on the preservation of the currently pristine natural environment of Kaieteur Falls.Keywords: nature tourism, GIS, Amazonia, national parks
Procedia PDF Downloads 16693 Catalytic Dehydrogenation of Formic Acid into H2/CO2 Gas: A Novel Approach
Authors: Ayman Hijazi, Witold Kwapinski, J. J. Leahy
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Finding a sustainable alternative energy to fossil fuel is an urgent need as various environmental challenges in the world arise. Therefore, formic acid (FA) decomposition has been an attractive field that lies at the center of biomass platform, comprising a potential pool of hydrogen energy that stands as a new energy vector. Liquid FA features considerable volumetric energy density of 6.4 MJ/L and a specific energy density of 5.3 MJ/Kg that qualifies it in the prime seat as an energy source for transportation infrastructure. Additionally, the increasing research interest in FA decomposition is driven by the need of in-situ H2 production, which plays a key role in the hydrogenation reactions of biomass into higher value components. It is reported elsewhere in literature that catalytic decomposition of FA is usually performed in poorly designed setup using simple glassware under magnetic stirring, thus demanding further energy investment to retain the used catalyst. it work suggests an approach that integrates designing a novel catalyst featuring magnetic property with a robust setup that minimizes experimental & measurement discrepancies. One of the most prominent active species for dehydrogenation/hydrogenation of biomass compounds is palladium. Accordingly, we investigate the potential of engrafting palladium metal onto functionalized magnetic nanoparticles as a heterogeneous catalyst to favor the production of CO-free H2 gas from FA. Using ordinary magnet to collect the spent catalyst renders core-shell magnetic nanoparticles as the backbone of the process. Catalytic experiments were performed in a jacketed batch reactor equipped with an overhead stirrer under inert medium. Through a novel approach, FA is charged into the reactor via high-pressure positive displacement pump at steady state conditions. The produced gas (H2+CO2) was measured by connecting the gas outlet to a measuring system based on the amount of the displaced water. The novelty of this work lies in designing a very responsive catalyst, pumping consistent amount of FA into a sealed reactor running at steady state mild temperatures, and continuous gas measurement, along with collecting the used catalyst without the need for centrifugation. Catalyst characterization using TEM, XRD, SEM, and CHN elemental analyzer provided us with details of catalyst preparation and facilitated new venues to alter the nanostructure of the catalyst framework. Consequently, the introduction of amine groups has led to appreciable improvements in terms of dispersion of the doped metals and eventually attaining nearly complete conversion (100%) of FA after 7 hours. The relative importance of the process parameters such as temperature (35-85°C), stirring speed (150-450rpm), catalyst loading (50-200mgr.), and Pd doping ratio (0.75-1.80wt.%) on gas yield was assessed by a Taguchi design-of-experiment based model. Experimental results showed that operating at lower temperature range (35-50°C) yielded more gas while the catalyst loading and Pd doping wt.% were found to be the most significant factors with a P-values 0.026 & 0.031, respectively.Keywords: formic acid decomposition, green catalysis, hydrogen, mesoporous silica, process optimization, nanoparticles
Procedia PDF Downloads 5292 Implementation of Real-World Learning Experiences in Teaching Courses of Medical Microbiology and Dietetics for Health Science Students
Authors: Miriam I. Jimenez-Perez, Mariana C. Orellana-Haro, Carolina Guzman-Brambila
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As part of microbiology and dietetics courses, students of medicine and nutrition analyze the main pathogenic microorganisms and perform dietary analyzes. The course of microbiology describes in a general way the main pathogens including bacteria, viruses, fungi, and parasites, as well as their interaction with the human species. We hypothesize that lack of practical application of the course causes the students not to find the value and the clinical application of it when in reality it is a matter of great importance for healthcare in our country. The courses of the medical microbiology and dietetics are mostly theoretical and only a few hours of laboratory practices. Therefore, it is necessary the incorporation of new innovative techniques that involve more practices and community fieldwork, real cases analysis and real-life situations. The purpose of this intervention was to incorporate real-world learning experiences in the instruction of medical microbiology and dietetics courses, in order to improve the learning process, understanding and the application in the field. During a period of 6 months, medicine and nutrition students worked in a community of urban poverty. We worked with 90 children between 4 and 6 years of age from low-income families with no access to medical services, to give an infectious diagnosis related to nutritional status in these children. We expect that this intervention would give a different kind of context to medical microbiology and dietetics students improving their learning process, applying their knowledge and laboratory practices to help a needed community. First, students learned basic skills in microbiology diagnosis test during laboratory sessions. Once, students acquired abilities to make biochemical probes and handle biological samples, they went to the community and took stool samples from children (with the corresponding informed consent). Students processed the samples in the laboratory, searching for enteropathogenic microorganism with RapID™ ONE system (Thermo Scientific™) and parasites using Willis and Malloy modified technique. Finally, they compared the results with the nutritional status of the children, previously measured by anthropometric indicators. The anthropometric results were interpreted by the OMS Anthro software (WHO, 2011). The microbiological result was interpreted by ERIC® Electronic RapID™ Code Compendium software and validated by a physician. The results were analyses of infectious outcomes and nutritional status. Related to fieldwork community learning experiences, our students improved their knowledge in microbiology and were capable of applying this knowledge in a real-life situation. They found this kind of learning useful when they translate theory to a real-life situation. For most of our students, this is their first contact as health caregivers with real population, and this contact is very important to help them understand the reality of many people in Mexico. In conclusion, real-world or fieldwork learning experiences empower our students to have a real and better understanding of how they can apply their knowledge in microbiology and dietetics and help a much- needed population, this is the kind of reality that many people live in our country.Keywords: real-world learning experiences, medical microbiology, dietetics, nutritional status, infectious status.
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