Search results for: liquid chromatography

Authors: G. R. Hashemi Tabar, G. R. Razmi, F. Mirshekar

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Echinococcus granulosus have ten strains from G1 to G9. Each strain is related to special intermediated host. The morphology, epidemiology, treatment and control in these strains are different. There are many morphological and molecular methods to differentiate of Echinococcus strains. However, using both methods were provided better information about identification of each strain. The aim of study was to identify Echinococcus granulosus strain of hydrated cysts in slaughtered sheep using morphological and molecular methods in Mashhad area. In the present study, the infected liver and lung with hydatid cysts were collected and transferred to laboratory. The hydatid cyst liquid was extracted and morphological characters of rostellar hook protosclocies were measured using micrometer ocular. The total length of large blade length of large hooks, total length of small and blade length of small hooks, and number of hooks per protoscolex were 23± 0.3μm, 11.7±0.5 μm, 19.3±1.1 μm,8±1.1 and 33.7±0.7 μm, respectively. In molecular section of the study, DNA each samples was extracted with MBST Kit and development of PCR using special primers (EgF, EgR) which amplify fragment of ITS1 gen. The PCR product was digested with Bsh1236I enzyme. Based on pattern of PCR-RLFP results (four band forming), G1, G2 and G3 strain of Echinococcus granulosus were obtained. Differentiation of three strains was done using sequencing analysis and G1 strain was diagnosed. The agreement between the molecular results with morphometric characters of rosetellar hook was confirmed the presence of G1 strain of Echinococcus in the slaughtered sheep of Mashhad area.

Keywords: Echinococcus granulosus, Hydatid cyst, PCR, sheep

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Authors: Funda Ates

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Primary energy sources, such as petroleum, coal and natural gas are principle responsible of world’s energy consumption. However, the rapid worldwide increase in the depletion of these energy sources is remarkable. In addition to this, they have damaging environmentally effect. Renewable energy sources are capable of providing a considerable fraction of World energy demand in this century. Biomass is one of the most abundant and utilized sources of renewable energy in the world. It can be converted into commercial fuels, suitable to substitute for fossil fuels. A high number of biomass types can be converted through thermochemical processes into solid, liquid or gaseous fuels. Pyrolysis is the thermal decomposition of biomass in the absence of air or oxygen. In this study, barley straw has been investigated as an alternative feedstock to obtain fuels and chemicals via pyrolysis in fixed-bed reactor. The influence of pyrolysis temperature in the range 450–750 °C as well as the catalyst effects on the products was investigated and the obtained results were compared. The results indicated that a maximum oil yield of 20.4% was obtained at a moderate temperature of 550 °C. Oil yield decreased by using catalyst. Pyrolysis oils were examined by using instrumental analysis and GC/MS. Analyses revealed that the pyrolysis oils were chemically very heterogeneous at all temperatures. It was determined that the most abundant compounds composing the bio-oil were phenolics. Catalyst decreased the reaction temperature. Most of the components obtained using a catalyst at moderate temperatures was close to those obtained at high temperatures without using a catalyst. Moreover, the use of a catalyst also decreased the amount of oxygenated compounds produced.

Keywords: Barley straw, pyrolysis, catalyst, phenolics

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Authors: Bernhard Widhalm, Cornelia Rieder-Gradinger, Ewald Srebotnik

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Pine wood (Pinus sylvestris L.) is the basic raw material for the production of Oriented Strand Boards (OSB) and the major source of volatile organic compounds, especially terpenes (like α- and β-pinene). To lower the total emission level of OSB, terpene metabolising microorganisms were therefore applied onto pine wood strands for the production of emission-reduced boards. Suitable microorganisms were identified during preliminary tests under laboratory conditions. At first, their terpene degrading potential was investigated in liquid culture, followed by laboratory tests using unsterile pine wood particles and strands. The main focus was laid on an adoptable terpene reduction in a short incubation time. An optimised bacterial mixture of Pseudomonas putida and Pseudomonas fluorescens showed the best results and was therefore used for further experiments on a larger scale. In an industry-compatible testing procedure, pine wood strands were incubated with the bacterial mixture for a period of 2 to 4 days. Incubation time was stopped by drying the strands. OSB were then manufactured from the pre-treated strands and emissions were measured by means of SPME/GC-MS analysis. Bacterial pre-treatment of strands resulted in a reduction of α-pinene- and β-pinene-emissions from OSB by 40% and 70%, respectively, even after only 2 days of incubation. The results of the investigation provide a basis for the application of microbial treatment within the industrial OSB production line, where shortest possible incubation times are required. For this purpose, the performance of the bacterial mixture will have to be further optimised.

Keywords: GC-MS, OSB, Pseudomonas sp., terpene degradation

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Authors: Ayman El Sabagh, AbdElhamid Omar, Dekoum Assaha, Khair Mohammad Youldash, Akihiro Ueda, Celaleddin Barutçular, Hirofumi Saneoka

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Drought and salinity stress are a worldwide problem, constraining global crop production seriously. Hence, soybean is susceptible to yield loss from water deficit and salinity stress. Therefore, different approaches have been suggested to solve these issues. Osmoprotectants play an important role in protection the plants from various environmental stresses. Moreover, organic fertilization has several beneficial effects on agricultural fields. Presently, efforts to maximize nitrogen fixation in soybean are critical because of widespread increase in soil degradation in Egypt. Therefore, a greenhouse research was conducted at plant nutritional physiology laboratory, Hiroshima University, Japan for assessing the impact of exogenous osmoregulators and compost application in alleviating the adverse effects of salinity and water stress on soybean. Treatments was included (i) water stress treatments (different soil moisture levels consisting of (100%, 75%, and 50% of field water holding capacity), (ii) salinity concentrations (0 and 15 mM) were applied in fully developed trifoliolate leaf node (V1), (iii) compost treatments (0 and 24 t ha-1) and (iv) the exogenous, proline and glycine betaine concentrations (0 mM and 25 mM) for each, was applied at two growth stages (V1 and R1). The seeds of soybean cultivar Giza 111, was sown into basin from wood (length10 meter, width 50cm, height 50cm and depth 350cm) containing a soil mixture of granite regosol soil and perlite (2:1 v/v). The nitrogen-fixing activity was estimated by using gas chromatography and all measurements were made in three replicates. The results showed that water deficit and salinity stress reduced biological nitrogen fixation and specific nodule activity than normal irrigation conditions. Exogenous osmoprotectants were improved biological nitrogen fixation and specific nodule activity as well as, applying of compost led to improving many of biological nitrogen fixation and specific nodule activity with superiority than stress conditions. The combined application compost fertilizer and exogenous osmoprotectants were more effective in alleviating the adverse effect of stress to improve biological nitrogen fixation and specific nodule activity of Soybean.

Keywords: a biotic stress, biological nitrogen fixation, compost, osmoprotectants, specific nodule activity, soybean

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Authors: Olufunmilola A. Abiodun, Adegbola O. Dauda, Ayobami Ojo, Samson A. Oyeyinka

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Serendipity berry (Dioscoreophyllum cumminsii diel) is a tropical dioecious rainforest vine and native to tropical Africa. The vine grows during the raining season and is used mainly as sweetener. The sweetener in the berry is known as monellin which is sweeter than sucrose. The sweetener is extracted from the fruits and the seed is discarded. The discarded seeds contain bitter principles but had high yield of oil. Serendipity oil was extracted using three methods (N-hexane, expression and expression/n-hexane). Fatty acids and physicochemical properties of the oil obtained were determined. The oil obtained was clear, liquid and have odour similar to hydrocarbon. The percentage oil yield was 38.59, 12.34 and 49.57% for hexane, expression and expression-hexane method respectively. The seed contained high percentage of oil especially using combination of expression and hexane. Low percentage of oil was obtained using expression method. The refractive index values obtained were 1.443, 1.442 and 1.478 for hexane, expression and expression-hexane methods respectively. Peroxide value obtained for expression-hexane was higher than those for hexane and expression. The viscosities of the oil were 125.8, 128.76 and 126.87 cm³/s for hexane, expression and expression-hexane methods respectively which showed that the oil from expression method was more viscous than the other oils. The major fatty acids in serendipity seed oil were oleic acid (62.81%), linoleic acid (22.65%), linolenic (6.11%), palmitic acid (5.67%), stearic acid (2.21%) in decreasing order. Oleic acid which is monounsaturated fatty acid had the highest value. Total unsaturated fatty acids were 91.574, 92.256 and 90.426% for hexane, expression, and expression-hexane respectively. Combination of expression and hexane for extraction of serendipity oil produced high yield of oil. The oil could be refined for food and non-food application.

Keywords: serendipity seed oil, expression method, fatty acid, hexane

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Authors: R. Vetrimurugan, Terry Lim, M. J. Goodson, R. Nagarajan

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This study investigates the cleaning performance of high intensity 360 kHz frequency on the removal of nano-dimensional and sub-micron particles from various surfaces, uniformity of the cleaning tank and run to run variation of cleaning process. The uniformity of the cleaning tank was measured by two different methods i.e 1. ppbTM meter and 2. Liquid Particle Counting (LPC) technique. In the second method, aluminium metal spacer components was placed at various locations of the cleaning tank (such as centre, top left corner, bottom left corner, top right corner, bottom right corner) and the resultant particles removed by 360 kHz frequency was measured. The result indicates that the energy was distributed more uniformly throughout the entire cleaning vessel even at the corners and edges of the tank when megasonic sweeping technology is applied. The result also shows that rinsing the parts with 360 kHz frequency at final rinse gives lower particle counts, hence higher cleaning efficiency as compared to other frequencies. When megasonic sweeping technology is applied each piezoelectric transducers will operate at their optimum resonant frequency and generates stronger acoustic cavitational force and higher acoustic streaming velocity. These combined forces are helping to enhance the particle removal and at the same time improve the overall cleaning performance. The multiple extractions study was also carried out for various frequencies to measure the cleaning potential and asymptote value.

Keywords: power distribution, megasonic sweeping, cavitation intensity, particle removal, laser particle counting, nano, submicron

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Authors: Esra Yel, Tabriz Aslanov, Merve Sogancioglu, Suheyla Kocaman, Gulnare Ahmetli

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The disposal of waste plastics has become a major worldwide environmental problem. Pyrolysis of waste plastics is one of the routes to waste minimization and recycling that has been gaining interest. In pyrolysis, the pyrolysed material is separated into gas, liquid (both are fuel) and solid (char) products. All fractions have utilities and economical value depending upon their characteristics. The first objective of this study is to determine the co-pyrolysis product fractions of waste HDPE- (high density polyethylene) and LDPE (low density polyethylene)-olive pomace (OP) and to determine the qualities of the solid product char. Chars obtained at 700 °C pyrolysis were used in biocomposite preparation as additive. As the second objective, the effects of char on biocomposite quality were investigated. Pyrolysis runs were performed at temperature 700 °C with heating rates of 5 °C/min. Biocomposites were prepared by mixing of chars with bisphenol-F type epoxy resin in various wt%. Biocomposite properties were determined by measuring electrical conductivity, surface hardness, Young’s modulus and tensile strength of the composites. The best electrical conductivity results were obtained with HDPE-OP char. For HDPE-OP char and LDPE-OP char, compared to neat epoxy, the tensile strength values of the composites increased by 102% and 78%, respectively, at 10% char dose. The hardness measurements showed similar results to the tensile tests, since there is a correlation between the hardness and the tensile strength.

Keywords: biocomposite, char, olive pomace, pyrolysis

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Authors: Nagalingam Arun Prasanth, Ahmed Syed Adnan, S. H. Yeo

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Surface topography plays a significant role in the functional performance of engineered parts. It is important to have a control on the surface geometry and understanding on the surface details to get the desired performance. Hence, in the current research contribution, a non-contact micro-texturing technique has been explored and developed. The technique involves ultrasonic excitation of a tool as a prime source of surface texturing for aluminum alloy workpieces. The specimen surface is polished first and is then immersed in a liquid bath containing 10% weight concentration of Ti6Al4V grade 5 spherical powders. A submerged slurry jet is used to recirculate the spherical powders under the ultrasonic horn which is excited at an ultrasonic frequency and amplitude of 40 kHz and 70 µm respectively. The distance between the horn and workpiece surface was remained fixed at 200 µm using a precision control stage. Texturing effects were investigated for different process timings of 1, 3 and 5 s. Thereafter, the specimens were cleaned in an ultrasonic bath for 5 mins to remove loose debris on the surface. The developed surfaces are characterized by optical and contact surface profiler. The optical microscopic images show a texture of circular spots on the workpiece surface indented by titanium spherical balls. Waviness patterns obtained from contact surface profiler supports the texturing effect produced from the proposed technique. Furthermore, water droplet tests were performed to show the efficacy of the proposed technique to develop hydrophilic surfaces and to quantify the texturing effect produced.

Keywords: surface texturing, surface modification, topography, ultrasonic

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Authors: Helen Dilman, Eyal Oz, Shmuel Hayun, Nahum Frage

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The conventional approach for manufacturing silicon carbide and boron carbide reaction bonded composites is based on infiltrating a ceramic porous preform with molten silicon. The relatively high melting temperature of the silicon infiltrating medium is a drawback of the process. The present contribution is concerned with an approach that allows obtaining reaction bonded composites by pressure-less infiltration at a significantly lower (850-1000oC) temperature range. This approach was applied for the fabrication of fully dense SiC/(Si-Al) and (SiC+B4C)/(Si-Al) composites. The key feature of the approach is based on using Si alloys with low melting temperature and the Mg-vapor atmosphere, under which an adequate wetting between ceramics and liquid alloys for the infiltration process is achieved. In the first set of the experiments ceramic performs compacted from multimodal SiC powders (with the green density of about 27 vol. %) without free carbon addition were infiltrated by Si-20%Al alloy at 950oC. In the second set, 19 vol. % of a fine boron carbide powder was added to SiC powders as a source of carbon. The green density of the SiC-B4C preforms was about 23-25 vol. %. In both cases, successful infiltration was achieved and the composites were fully dense. The density of the composites was about 3g/cm3. For the SiC based composites the hardness value was 750±150HV, Young modulus-280GPa and bending strength-240±30MPa. These values for (SiC-B4C)/(Si-Al) composites (1460±200HV, 317GPa and 360±20MPa) were significantly higher due to the formation of novel ceramics phases. Microstructural characteristics of the composites and their phase composition will be discussed.

Keywords: boron carbide, composites, infiltration, low temperatures, silicon carbide

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Authors: S. Djaziri, F. Sket, A. Hynowska, S. Milenkovic

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The use of Fe-Al based intermetallics as an alternative to Cr/Ni based stainless steels is very promising for industrial applications that use critical raw materials parts under extreme conditions. However, the development of advanced Fe-Al based intermetallics with appropriate mechanical properties presents several challenges that involve appropriate processing and microstructure control. A processing strategy is being developed which aims at producing a net-shape porous Fe-based preform that is infiltrated with molten Al or Al-alloy. In the present work, porous Fe-based preforms produced by two different methods (selective laser melting (SLM) and Kochanek-process (KE)) are studied during infiltration with molten aluminum. In the objective to elucidate the mechanisms underlying the formation of Fe-Al intermetallic phases during infiltration, an in-house furnace has been designed for in situ observation of infiltration at synchrotron facilities combining x-ray radiography (XR) and x-ray diffraction (XRD) techniques. The feasibility of this approach has been demonstrated, and information about the melt flow front propagation has been obtained. In addition, reactive infiltration has been achieved where a bi-phased intermetallic layer has been identified to be formed between the solid Fe and liquid Al. In particular, a tongue-like Fe₂Al₅ phase adhering to the Fe and a needle-like Fe₄Al₁₃ phase adhering to the Al were observed. The growth of the intermetallic compound was found to be dependent on the temperature gradient present along the preform as well as on the reaction time which will be discussed in view of the different obtained results.

Keywords: combined synchrotron radiography and diffraction, Fe-Al intermetallic compounds, in-situ molten Al infiltration, porous solid Fe preforms

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Authors: Shaya Mahmoudian, Mohammad Reza Sazegar, Nazanin Afshari

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Graphene, a single layer of carbon atoms in a hexagonal lattice, has recently attracted great attention due to its unique mechanical, thermal and electrical properties. The high aspect ratio and unique surface features of graphene resulted in significant improvements of the nano composites properties. In this study, nano composite fibres made of cellulose and graphene nano platelets were wet spun from solution by using ionic liquid, 1-ethyl-3-methylimidazolium acetate (EMIMAc) as solvent. The effect of graphene loading on the thermal and electrical properties of the nanocomposite fibres was investigated. The nano composite fibres characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) analysis. XRD analysis revealed a cellulose II crystalline structure for regenerated cellulose and the nano composite fibres. SEM images showed a homogenous morphology and round cross section for the nano composite fibres along with well dispersion of graphene nano platelets in regenerated cellulose matrix. The incorporation of graphene into cellulose matrix generated electrical conductivity. At 6 wt. % of graphene, the electrical conductivity was 4.7 × 10-4 S/cm. The nano composite fibres also showed considerable improvements in thermal stability and char yield compared to pure regenerated cellulose fibres. This work provides a facile and environmentally friendly method of preparing nano composite fibres based on cellulose and graphene nano platelets that can find several applications in cellulose-based carbon fibres, conductive fibres, apparel, etc.

Keywords: nanocomposite, graphene nanoplatelets, regenerated cellulose, electrical properties

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Authors: Yaseen Muhammad, Zhenxia Zhao, Zhangfa Tong

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A key problem with hydrodesulfurization (HDS) process of fuel oils is the application of severe operating conditions. In this study, we proposed the catalytic HDS of dibenzothiophene (DBT) integrated with ionic liquids (ILs) application at mild temperature and pressure over low loaded (0.5 wt.%) Pd promoted Co-Mo@Al₂O₃ and Ni-Mo@Al₂O₃ catalysts. Among the thirteen ILs tested, [BMIM]BF₄, [(CH₃)₄N]Cl, [EMIM]AlCl₄, and [(C₈H₁₇)(C₃H₇)₃P]Br enhanced the catalytic HDS efficiency while the latest ranked the top of activity list as confirmed by DFT studies as well. Experimental results revealed that Pd incorporation greatly enhanced the HDS activity of classical Co or Ni based catalysts. At mild optimized experimental conditions of 1 MPa H₂ pressure, 120 oC, IL:oil ratio of 1:3 and 4 h reaction time, the % DBT conversion (21 %) by Ni-Mo@Al₂O₃ was enhanced to 69 % (over Pd-Ni-Mo@ Al₂O₃) using [(C₈H₁₇) (C₃H₇)₃P]Br. The fresh and spent catalysts were characterized for textural properties using XPS, SEM, EDX, XRD and BET surface area techniques. An overall catalytic HDS activity followed the order of: Pd-Ni-Mo@Al₂O₃ > Pd-Co-Mo@Al₂O₃ > Ni-Mo@Al₂O₃ > Co-Mo@Al₂O₃. [(C₈H₁₇) (C₃H₇)₃P]Br.could be recycled four times with minimal decrease in HDS activity. Reaction products were analyzed by GC-MS which helped in proposing reaction mechanism for the IL coupled HDS process. The present approach attributed to its cost-effective nature, ease of operation with less mechanical requirements in terms of mild operating conditions, and high efficiency could be deemed as an alternative approach for the HDS of DBT on industrial level applications.

Keywords: DFT simulation, GC-MS and reaction mechanism, Ionic liquid coupled HDS of DBT, low Pd loaded catalyst, mild operating condition

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Authors: S. Visetpotjanakit, C. Khrautongkieo

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Radiation monitoring in the environment and foodstuffs is one of the main responsibilities of Office of Atoms for Peace (OAP) as the nuclear regulatory body of Thailand. The main goal of the OAP is to assure the safety of the Thai people and environment from any radiological incidents. Various radioanalytical methods have been developed to monitor radiation and radionuclides in the environmental and foodstuff samples. To validate our analytical performance, several proficiency test exercises from the International Atomic Energy Agency (IAEA) have been performed. Here, the results of a proficiency test exercise referred to as the Proficiency Test for Tritium, Cobalt, Strontium and Caesium Isotopes in Seawater 2017 (IAEA-RML-2017-01) are presented. All radionuclides excepting ³H were analysed using various radioanalytical methods, i.e. direct gamma-ray counting for determining ⁶⁰Co, ¹³⁴Cs and ¹³⁷Cs and developed radiochemical techniques for analysing ¹³⁴Cs, ¹³⁷Cs using AMP pre-concentration technique and 90Sr using di-(2-ethylhexyl) phosphoric acid (HDEHP) liquid extraction technique. The analysis results were submitted to IAEA. All results passed IAEA criteria, i.e. accuracy, precision and trueness and obtained ‘Accepted’ statuses. These confirm the data quality from the OAP environmental radiation laboratory to monitor radiation in the environment.

Keywords: international atomic energy agency, proficiency test, radiation monitoring, seawater

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Authors: Young Jae Choung, Seoung Bum Kim

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Statistical Process Control (SPC) is a popular technique for process monitoring. A widely used tool in SPC is a control chart, which is used to detect the abnormal status of a process and maintain the controlled status of the process. Traditional control charts, such as Hotelling’s T2 control chart, are effective techniques to detect abnormal observations and monitor processes. However, many complicated manufacturing systems exhibit nonlinearity because of the different demands of the market. In this case, the unregulated use of a traditional linear modeling approach may not be effective. In reality, many industrial processes contain the nonlinear and time-varying properties because of the fluctuation of process raw materials, slowing shift of the set points, aging of the main process components, seasoning effects, and catalyst deactivation. The use of traditional SPC techniques with time-varying data will degrade the performance of the monitoring scheme. To address these issues, in the present study, we propose a parameterless self-organizing map (PLSOM)-based control chart. The PLSOM-based control chart not only can manage a situation where the distribution or parameter of the target observations changes, but also address the nonlinearity of modern manufacturing systems. The control limits of the proposed PLSOM chart are established by estimating the empirical level of significance on the percentile using a bootstrap method. Experimental results with simulated data and actual process data from a thin-film transistor-liquid crystal display process demonstrated the effectiveness and usefulness of the proposed chart.

Keywords: control chart, parameter-less self-organizing map, self-organizing map, time-varying property

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Authors: Zafar Iqbal, Sana Irshad Khan

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Organic extract from newly isolated plant growth promoting fungus (PGPF) Penicillium sp EU0013 was subjected to bioassays including anti fungal (disc diffusion) cytotoxicity (brine shrimp lethality), herbicidal (Lemna minor) and nematicidal activities. Metabolite profile of the extract was also assessed using HPLC analysis with the aim to identify bioactive natural products in the extract as new drug candidate(s). The extract showed anti fungal potential against tested fungal pathogens. Growth of the Wilt pathogen Fusarium oxyosproum was inhibited up to 63% when compared to negative reference. Activity against brine shrimps was weak and mortality up to 10% was observed at concentration of 200 µg. mL-1. The extract exhibited no toxicity against Lemna minor frond at 200 µg. mL-1. Nematicidal activity was observed very potent against root knot nematode and LC50 value was calculated as 52.5 ug. mL-1 using probit analysis. Methodically assessment of metabolites profile by HPLC showed the presence of kojic acid (Rt 1.4 min) and aflatoxin B1 (Rt 5.9 min) in the mycellial extract as compared with standards. The major unidentified metabolite was eluted at Rt 8.6 along with other minor peaks. The observed high toxicity against root knot nematode was attributed to the unidentified compounds that make fungal extract worthy of further exploration for isolation and structural characterization studies for development of future commercial nematicidal compound(s).

Keywords: penicillium, nematicidal activity, metabolites, HPLC

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Authors: Nahal Najafi, Afsaneh Yegdaneh, Sedigheh Saberi

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Introduction: Leishmaniasis poses a substantial global risk, affecting millions and resulting in thousands of cases each year in endemic regions. Challenges in current leishmaniasis treatments include drug resistance, high toxicity, and pancreatitis. Marine compounds, particularly brown algae, serve as a valuable source of inspiration for discovering treatments against Leishmania. Material and method: Padina pavonica was collected from the Persian Gulf. The seaweeds were dried and extracted with methanol: ethylacetate (1:1). The extract was partitioned to hexane (Hex), dicholoromethane (DCM), butanol, and water by Kupchan partitioning method. Hex partition was fractionated by silica gel column chromatography to 10 fractions (Fr. 1-10). Fr. 6 was further separated by the normal phase HPLC method to yield compounds 1-3. The structures of isolated compounds were elucidated by NMR, Mass, and other spectroscopic methods. Hex and DCM partitions, Fr. 6 and compounds 1-3, were tested for anti-leishmanicidal activity. RAW cell lines were cultured in enriched RPMI (10% FBS, 1% pen-strep) in a 37°C CO2 5% incubator, while promastigote cells were initially cultured in NNN culture and subsequently transferred to the aforementioned medium. Cytotoxicity was assessed using MTT tests, anti-promastigote activity was evaluated through Hemocytometer chamber promastigote counting, and the impact of amastigote damage was determined by counting amastigotes within 100 macrophages. Results: NMR and Mass identified isolated compounds as fucosterol and two sulfoquinovosyldiacylglycerols (SQDG). Among the samples tested, Fr.6 exhibited the highest cytotoxicity (CC50=60.24), while compound 2 showed the lowest cytotoxicity (CC50=21984). Compound 1 and dichloromethane fraction demonstrated the highest and lowest anti-promastigote activity (IC50=115.7, IC50=16.42, respectively), and compound 1 and hexane fraction exhibited the highest and lowest anti-amastigote activity (IC50=7.874, IC50=40.18, respectively). Conclusion: All six samples, including Hex and DCM partitions, Fr.6, and compounds 1-3, demonstrate a noteworthy correlation between rising concentration and time, with a statistically significant P-value of ≤0.05. Considering the higher selectivity index of compound 2 compared to others, it can be inferred that the presence of sulfur groups and unsaturated chains potentially contributes to these effects by impeding the DNA polymerase, which, of course, needs more research.

Keywords: Padina, leishmania, sulfoquinovosyldiacylglycerol, cytotoxicity

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Authors: Houria Rezala, Jose Luis Valverde, Amaya Romero, Alessandra Molinari, Andrea Maldotti

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A pillared montmorillonite containing iron doped titania (Fe/Ti-PILC) has been prepared from a natural clay. This material has been characterized by X-ray diffraction, nitrogen adsorption, temperature programmed desorption of ammonia, inductively coupled plasma atomic emission spectroscopy, atomic absorption, and diffuse reflectance UV-VIS spectroscopy. The layer structure of Fe/Ti-PILC resulted to be ordered with an insertion of pillars, which caused a slight increase in the basal spacing of the clay. Its specific surface area was about three times larger than that of the parent Na-montmorillonite due principally to the creation of a remarkable microporous network. The doped material was a robust photocatalyst able to oxidize liquid alkyl aromatics to the corresponding carbonylic derivatives, using O2 as the oxidizing species, at mild pressure and temperature conditions. Accumulation of valuable carbonylic derivatives was possible since their over-oxidation to carbon dioxide was negligible. Fe/Ti-PILC was able to discriminate between toluene and cyclohexane in favor of the aromatic compound with an efficiency that is about three times higher than that of titanium pillared clays (Ti-PILC). It is likely that the addition of iron favored the formation of new acid sites able to interact with the aromatic substrate. Iron doping caused a significant TiO2 visible light-induced activity (wavelength > 400 nm) with only minor negative effects on its performance under UV-light irradiation (wavelength > 290 nm).

Keywords: alkyl aromatics oxidation, heterogeneous photocatalysis, iron doping, pillared clays

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Authors: Brayan Daniel Riascos Arteaga, Carlos Costa Perez

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Landfill leachate has an important fraction of humic substances, mainly humic acids (HAs), which often represent more than half value of COD, specially in liquids proceeded from composting processes of organic fraction of solid wastes. We propose in this article a new method of pH sensitive flocculation for COD and colour reduction in landfill leachate based on the chemical properties of HAs. Landfill leachate with a high content of humic acids can be efficiently treated by pH sensitive flocculation at pH 2.0, reducing COD value in 86.1% and colour in 84.7%. Mechanism of pH sensitive flocculation is based in protonation first of phenolic groups and later of carboxylic acid groups in the HAs molecules, resulting in a reduction of Zeta potential value. For pH over neutrality, carboxylic acid and phenolic groups are ionized and Zeta potential increases in absolute value, maintaining HAs in suspension as colloids and conducting flocculation to be obstructed. Ionized anionic groups (carboxylates) can interact electrostatically with cations abundant in leachate (site binding) aiding to maintain HAs in suspension. Simulation of this situation and ideal visualization of Zeta potential behavior is described in the paper and aggregation of molecules by H-bonds is proposed as the main step in separation of HAs from leachate and reduction of COD value in this complex liquid. CHNS analysis, FT-IR spectrometry and UV–VIS spectrophotometry show chemical elements content in the range of natural and commercial HAs, clear aromaticity and carboxylic acids and phenolic groups presence in the precipitate from landfill leachate

Keywords: landfill leachate, humic acids, COD, chemical treatment, flocculation

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Authors: Mohamed Youssry, Dominique Guyomard, Bernard Lestriez

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In order to enhance the energy and power densities of electrodes for energy storage systems, the formulation and processing of electrode slurries proved to be a critical issue in determining the electrode performance. In this study, we introduce novel approach to formulate carbon black slurries based on microemulsion and lyotropic liquid crystalline phases (namely, lamellar phase) composed of non-ionic surfactant (Triton X100), decanol and water. Simultaneous measurements of electrical properties of slurries under shear flow (rheology) have been conducted to elucidate the microstructure evolution with the surfactant concentration and decanol/water ratio at rest, as well as, the structural transition under steady-shear which has been confirmed by rheo-microscopy. Interestingly, the carbon black slurries at low decanol/water ratio are weak-gel (flowable) with higher electrical conductivity than those at higher ratio which behave strong-gel viscoelastic response. In addition, the slurries show recoverable electrical behaviour under shear flow in tandem with the viscosity trend. It is likely that oil-in-water microemulsion enhances slurries’ stability without affecting on the percolating network of carbon black. On the other hand, the oil-in-water analogous and bilayer structure of lamellar phase cause the slurries less conductive as a consequence of losing the network percolation. These findings are encouraging to formulate microemulsion-based electrodes for energy storage system (lithium-ion batteries).

Keywords: electrode slurries, microemulsion, microstructure transition, rheo-electrical properties

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Authors: Bruno Baptista, Andreia S. R. Oliveira, Patricia V. Mendonca, Jorge F. J. Coelho, Fani Sousa

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Drug delivery systems are designed to allow adequate protection and controlled delivery of drugs to specific locations. These systems aim to reduce side effects and control the biodistribution profile of drugs, thus improving therapeutic efficacy. This study involved the synthesis of polymeric nanoparticles, based on amphiphilic diblock copolymers, comprising a biocompatible, poly (oligo (ethylene oxide) methyl ether methacrylate (POEOMA) as hydrophilic segment and a pH-sensitive block, the poly (2-diisopropylamino)ethyl methacrylate) (PDPA). The objective of this work was the development of polymeric pH-responsive nanoparticles to encapsulate and carry small RNAs as a model to further develop non-coding RNAs delivery systems with therapeutic value. The responsiveness of PDPA to pH allows the electrostatic interaction of these copolymers with nucleic acids at acidic pH, as a result of the protonation of the tertiary amine groups of this polymer at pH values below its pKa (around 6.2). Initially, the molecular weight parameters and chemical structure of the block copolymers were determined by size exclusion chromatography (SEC) and nuclear magnetic resonance (1H-NMR) spectroscopy, respectively. Then, the complexation with small RNAs was verified, generating polyplexes with sizes ranging from 300 to 600 nm and with encapsulation efficiencies around 80%, depending on the molecular weight of the polymers, their composition, and concentration used. The effect of pH on the morphology of nanoparticles was evaluated by scanning electron microscopy (SEM) being verified that at higher pH values, particles tend to lose their spherical shape. Since this work aims to develop systems for the delivery of non-coding RNAs, studies on RNA protection (contact with RNase, FBS, and Trypsin) and cell viability were also carried out. It was found that they induce some protection against constituents of the cellular environment and have no cellular toxicity. In summary, this research work contributes to the development of pH-sensitive polymers, capable of protecting and encapsulating RNA, in a relatively simple and efficient manner, to further be applied on drug delivery to specific sites where pH may have a critical role, as it can occur in several cancer environments.

Keywords: drug delivery systems, pH-responsive polymers, POEOMA-b-PDPA, small RNAs

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Authors: Inci Develioglu, Hasan Firat Pulat

Abstract:

Sustainable development is one of the most important topics in today's world, and it is also an important research topic for geoenvironmental engineering. Dredging process is performed to expand the river and port channel, flood control and accessing harbors. Every year large amount of sediment are dredged for these purposes. Dredged marine soils can be reused as filling materials, road and foundation embankments, construction materials and wildlife habitat developments. In this study, geotechnical engineering properties and compressibility behavior of dredged soil obtained from the Izmir Bay were investigated. The samples with four different organic matter contents were obtained and particle size distributions, consistency limits, pH and specific gravity tests were performed. The consolidation tests were conducted to examine organic matter content (OMC) effects on compressibility behavior of dredged soil. This study has shown that the OMC has an important effect on the engineering properties of dredged soils. The liquid and plastic limits increased with increasing OMC. The lowest specific gravity belonged to sample which has the maximum OMC. The specific gravity values ranged between 2.76 and 2.52. The maximum void ratio difference belongs to sample with the highest OMC (De_11% = 0.38)_. As the organic matter content of the samples increases, the change in the void ratio has also increased. The compression index increases with increasing OMC.

Keywords: compressibility, consolidation, geotechnical properties, organic matter content, dredged soil

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Authors: M. Agostini, A. Sonato, G. Greco, M. Travagliati, G. Ruffato, E. Gazzola, D. Liuni, F. Romanato, M. Cecchini

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Since their first demonstration in the early 1980s, surface plasmon resonance (SPR) sensors have been widely recognized as useful tools for detecting chemical and biological species, and the interest of the scientific community toward this technology has known a rapid growth in the past two decades owing to their high sensitivity, label-free operation and possibility of real-time detection. Recent works have suggested that a turning point in SPR sensor research would be the combination of SPR strategies with other technologies in order to reduce human handling of samples, improve integration and plasmonic sensitivity. In this light, microfluidics has been attracting growing interest. By properly designing microfluidic biochips it is possible to miniaturize the analyte-sensitive areas with an overall reduction of the chip dimension, reduce the liquid reagents and sample volume, improve automation, and increase the number of experiments in a single biochip by multiplexing approaches. However, as the fluidic channel dimensions approach the micron scale, laminar flows become dominant owing to the low Reynolds numbers that typically characterize microfluidics. In these environments mixing times are usually dominated by diffusion, which can be prohibitively long and lead to long-lasting biochemistry experiments. An elegant method to overcome these issues is to actively perturb the liquid laminar flow by exploiting surface acoustic waves (SAWs). With this work, we demonstrate a new approach for SPR biosensing based on the combination of microfluidics, SAW-induced mixing and the real-time phase-interrogation grating-coupling SPR technology. On a single lithium niobate (LN) substrate the nanostructured SPR sensing areas, interdigital transducer (IDT) for SAW generation and polydimethylsiloxane (PDMS) microfluidic chambers were fabricated. SAWs, impinging on the microfluidic chamber, generate acoustic streaming inside the fluid, leading to chaotic advection and thus improved fluid mixing, whilst analytes binding detection is made via SPR method based on SPP excitation via gold metallic grating upon azimuthal orientation and phase interrogation. Our device has been fully characterized in order to separate for the very first time the unwanted SAW heating effect with respect to the fluid stirring inside the microchamber that affect the molecules binding dynamics. Avidin/biotin assay and thiol-polyethylene glycol (bPEG-SH) were exploited as model biological interaction and non-fouling layer respectively. Biosensing kinetics time reduction with SAW-enhanced mixing resulted in a ≈ 82% improvement for bPEG-SH adsorption onto gold and ≈ 24% for avidin/biotin binding—≈ 50% and 18% respectively compared to the heating only condition. These results demonstrate that our biochip can significantly reduce the duration of bioreactions that usually require long times (e.g., PEG-based sensing layer, low concentration analyte detection). The sensing architecture here proposed represents a new promising technology satisfying the major biosensing requirements: scalability and high throughput capabilities. The detection system size and biochip dimension could be further reduced and integrated; in addition, the possibility of reducing biological experiment duration via SAW-driven active mixing and developing multiplexing platforms for parallel real-time sensing could be easily combined. In general, the technology reported in this study can be straightforwardly adapted to a great number of biological system and sensing geometry.

Keywords: biosensor, microfluidics, surface acoustic wave, surface plasmon resonance

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Authors: Tinomuvonga Manenji Zhou, Eubert Mahofa, Tatenda Crispen Madzokere

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The nanostructured formulation can increase fertilizer efficacy and uptake ratio of the soil nutrients in agriculture production and save fertilizer resources. Controlled release modes have properties of both release rate and release pattern of nutrients, for fertilizers that are soluble in water might be correctly controlled. Nanoparticles can reduce the rate at which fertilizer nutrients are in the soil by leaching. A slow release NPK-hydroxyapatite nano hybrid fertilizer was synthesized using exfoliated bentonite as filler material. A simple, scalable method was used to synthesize the nitrogen-phosphorus hydroxyapatite nano fertilizer, where calcium hydroxide, phosphoric acid, and urea were used as precursor material, followed by the incorporation of potassium through a liquid grinding method. The product obtained was an NPK-hydroxyapatite nano hybrid fertilizer. A quantitative analysis was done to determine the percentage of nitrogen, phosphorus, and potassium in the hybrid fertilizer. AAS was used to determine the percentage of potassium in the fertilizer. An accelerated water test was conducted to compare the nutrient release behavior of nutrients between the synthesized NPK-hydroxyapatite nano hybrid fertilizer and commercial NPK fertilizer. The rate of release of Nitrogen, phosphorus, and potassium was significantly lower in the synthesized NPK hydroxyapatite nano hybrid fertilizer than in the convectional NPK fertilizer. The synthesized fertilizer was characterized using XRD. NPK hydroxyapatite nano hybrid fertilizer encapsulated in exfoliated bentonite thus prepared can be used as an environmentally friendly fertilizer formulation which could be extended to solve one of the major problems faced in the global fertilization of low nitrogen, phosphorus, and potassium use efficiency in agriculture.

Keywords: NPK hydroxyapatite nano hybrid fertilizer, bentonite, encapsulation, low release

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Authors: Lourdita Llanto

Abstract:

This study described and assessed the implementation of artificial insemination (AI) for cattle and carabaos in the Bicol Region, Philippines: Albay, Sorsogon and Camarines Sur. Three hundred respondents were interviewed. Results were analyzed using frequency counts, means, percentages and chi-square test. Semen samples from different stations were analyzed for motility, viability and morphology. T-test was used in semen quality evaluation. Provincial AI coordinators (PAIC) were male, averaging 59 years old, married, had college education, served in government service for 34 years, but as PAIC for 5.7 years. All had other designations. Mean AI operation was 11.33 years with annual support from the local government unit of Php76,666.67. AI technicians were males, married, with college education, and trained on AI. Problems were on mobility; inadequate knowledge of farmers in animal raising and AI; and lack of liquid nitrogen and frozen semen supply. There was 2.95 municipalities and breedable cattle/carabaos of 3,091.25 per AI technician. Mean number of artificially inseminated animals per AI technician for 2011 was 28.57 heads for carabaos and 8.64 heads for cattle. There was very low participation rate among farmers. Carabaos were 6.52 years with parity 1.53. Cattle were 5.61 years, with parity of 1.51. Semen quality significantly (p ≤ 0.05) deteriorated in normal and live sperm with storage and handling at the provincial and field stations. Breed, AI technicians practices and AI operation significantly affected conception rate. Mean conception rate was 57.62%.

Keywords: artificial insemination, carabao, parity, mother tanks, frozen semen

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Authors: Ahmed Auda, Manjree Agarwala, Giles Hardya, Yonglin Rena

Abstract:

Botrytis cinerea is a major pest for many plants. It can attack a wide range of plant parts. It can attack buds, flowers, and leaves, stems, and fruit. However, B. cinerea can be mixed with other diseases that cause the same damage. There are many species of botrytis and more than one different strains of each. Botrytis might infect the foliage of nursery stock stored through winter in damp conditions. There are no known resistant plants. Botrytis must have nutrients or food source before it infests the plant. Nutrients leaking from wounded plant parts or dying tissue like old flower petals give the required nutrients. From this food, the fungus becomes more attackers and invades healthy tissue. Dark to light brown rot forms in the ill tissue. High humidity conditions support the growth of this fungus. However, we suppose that selection pressure can act on the morphological and neurophysiologic filter properties of the receiver and on both the biochemical and the physiological regulation of the signal. Communication is implied when signal and receiver evolves toward more and more specific matching, culminating. In other hand, receivers respond to portions of a body odor bouquet which is released to the environment not as an (intentional) signal but as an unavoidable consequence of metabolic activity or tissue damage. Each year Botrytis species can cause considerable economic losses to plant crops. Even with the application of strict quarantine and control measures, these fungi can still find their way into crops and cause the imposition of onerous restrictions on exports. Blueberry fruit mould caused by a fungal infection usually results in major losses during post-harvest storage. Therefore, the management of infection in early stages of disease development is necessary to minimize losses. The overall purpose of this study will develop sensitive, cheap, quick and robust diagnostic techniques for the detection of B. cinerea in blueberry. The specific aim was designed to investigate the performance of volatile organic compounds (VOCs) in the detection and discrimination of blueberry fruits infected by fungal pathogens with an emphasis on Botrytis in the early storage stage of post-harvest.

Keywords: botrytis cinerea, blueberry, GC/MS, VOCs

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Authors: Vladimir Viktorov, M. Readul Mahmud, Carmen Visconte

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Numerical simulation were carried out to study the mixing of miscible liquid at different inlets volume ratio (1 to 3) within two existing mixers namely Chain, Tear-drop and one new “C-H” mixer. The new passive C-H micromixer is developed based on split and recombine principles, combining the operation concepts of known Chain mixer and H mixer. The mixing performances of the three micromixers were predicted by a preliminary numerical analysis of the flow patterns inside the channel in terms of the segregation or distribution of path lines. Afterward, the efficiency and the pressure drop were investigated numerically, taking into account species transport. All numerical calculations were computed at a wide range of Reynolds number from 1 to 100. Among the presented three micromixers, tear-drop provides fairly good efficiency except in the middle range of Re numbers but has high-pressure drop. In addition, inlets flow ratio has a significant influence on efficiency, especially at the Re number range of 10 to 50, Moreover maximum increase of efficiency is almost 10% when inlets flow ratio is increased by 1. Chain mixer presents relatively low mixing efficiency at low and middle range of Re numbers (5≤Re≤50) but has reasonable pressure drop. Furthermore, Chain mixer shows almost no dependence on inlets flow ratio. Whereas, C-H mixer poses excellent mixing efficiency (more than 93%) for all range of Re numbers and causes the lowest pressure drop, On top of that efficiency has slight dependency on inlets flow ratio. In addition, C-H mixer shows respectively about three and two times lower pressure drop than Tear-drop and Chain mixers.

Keywords: CFD, micromixing, passive micromixer, SAR

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Authors: Rabitah Zakaria, Chan M. Luan, Nor Hakimah Ramly

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The extraction of vegetable oil using aqueous extraction process assisted by ionic extended surfactant has been investigated as an alternative to hexane extraction. However, the ionic extended surfactant has not been commercialised and its safety with respect to food processing is uncertain. Hence, food-grade non-ionic surfactants (Tween 20, Span 20, and Span 80) were proposed for the extraction of residual oil from palm-pressed mesocarp fibre. Palm-pressed mesocarp fibre contains a significant amount of residual oil ( 5-10 wt %) and its recovery is beneficial as the oil contains much higher content of vitamin E, carotenoids, and sterols compared to crude palm oil. In this study, the formulation of food-grade surfactants using a combination of high hydrophilic-lipophilic balance (HLB) surfactants and low HLB surfactants to produce micro-emulsion with very low interfacial tension (IFT) was investigated. The suitable surfactant formulation was used in the oil extraction process and the efficiency of the extraction was correlated with the IFT, droplet size and viscosity. It was found that a ternary surfactant mixture with a HLB value of 15 (82% Tween 20, 12% Span 20 and 6% Span 80) was able to produce micro-emulsion with very low IFT compared to other HLB combinations. Results suggested that the IFT and droplet size highly affect the oil recovery efficiency. Finally, optimization of the operating parameters shows that the highest extraction efficiency of 78% was achieved at 1:31 solid to liquid ratio, 2 wt % surfactant solution, temperature of 50˚C, and 50 minutes contact time.

Keywords: food-grade surfactants, aqueous extraction of residual oil, palm-pressed mesocarp fibre, interfacial tension

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Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

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A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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Authors: Akhila Konala, Jagadeeswara Reddy Vennapusa, Sujay Chattopadhyay

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The energy consumption of humankind is growing day by day due to an increase in the population, industrialization and their needs for living. Fossil fuels are the major source of energy to satisfy energy needs, which are non-renewable energy resources. So, there is a need to develop green resources for energy production and storage. Phase change materials (PCMs) derived from plants (green resources) are well known for their capacity to store the thermal energy as latent heat during their phase change from solid to liquid. This property of PCM could be used for storage of thermal energy. In this study, a composite with fatty acid (caprylic acid; M.P 15°C, Enthalpy 179kJ/kg) as a phase change material and expanded perlite as support porous matrix was prepared through direct impregnation method for thermal energy storage applications. The prepared composite was characterized using Differential scanning calorimetry (DSC), Field Emission Scanning Electron Microscope (FESEM), Thermal Gravimetric Analysis (TGA), and Fourier Transform Infrared (FTIR) spectrometer. The melting point of the prepared composite was 15.65°C, and the melting enthalpy was 82kJ/kg. The surface nature of the perlite was observed through FESEM. It was observed that there are micro size pores in the perlite surface, which were responsible for the absorption of PCM into perlite. In TGA thermogram, the PCM loss from composite was started at ~90°C. FTIR curves proved there was no chemical interaction between the perlite and caprylic acid. So, the PCM composite prepared in this work could be effective to use in temperature maintenance of buildings.

Keywords: caprylic acid, composite, phase change materials, PCM, perlite, thermal energy

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Authors: S. Rostampour Yasouri, M. Ghane, M. Doudi

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Leptospirosis is one of the most significant infectious and zoonotic diseases along with global spreading. This disease is causative agent of economoic losses and human fatalities in various countries, including Northern provinces of Iran. The aim of this research is to identify and compare the rapid diagnostic techniques of pathogenic leptospiras, considering the multifacetedness of the disease from a clinical manifestation and premature death of patients. In the spring and summer of 2020-2022, 25 fecal samples were collected from suspected leptospirosis patients and 25 Fecal samples from mice residing in the rice fields and factories in Tonekabon city. Samples were prepared by centrifugation and passing through membrane filters. Culture technique was used in liquid and solid EMJH media during one month of incubation at 30°C. Then, the media were examined microscopically. DNA extraction was conducted by extraction Kit. Diagnosis of leptospiras was enforced by PCR and Real time PCR (SYBR Green) techniques using lipL32 specific primer. Out of the patients, 11 samples (44%) and 8 samples (32%) were determined to be pathogenic Leptospira by Real time PCR and PCR technique, respectively. Out of the mice, 9 Samples (36%) and 3 samples (12%) were determined to be pathogenic Leptospira by the mentioned techniques, respectively. Although the culture technique is considered to be the gold standard technique, but due to the slow growth of pathogenic Leptospira and lack of colony formation of some species, it is not a fast technique. Real time PCR allowed rapid diagnosis with much higher accuracy compared to PCR because PCR could not completely identify samples with lower microbial load.

Keywords: culture, pathogenic leptospiras, PCR, real time PCR

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