Search results for: laboratory incubation

Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

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The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability

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Authors: Martin Pumera

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Graphene materials are being widely used in electrochemistry due to their versatility and excellent properties as platforms for biosensing. Here we present current trends in the electrochemical biosensing of pesticides and other toxic compounds. We explore two fundamentally different designs, (i) using graphene and other 2-D nanomaterials as an electrochemical platform and (ii) using these nanomaterials in the laboratory on chip design, together with paramagnetic beads. More specifically: (i) We explore graphene as transducer platform with very good conductivity, large surface area, and fast heterogeneous electron transfer for the biosensing. We will present the comparison of these materials and of the immobilization techniques. (ii) We present use of the graphene in the laboratory on chip systems. Laboratory on the chip had a huge advantage due to small footprint, fast analysis times and sample handling. We will show the application of these systems for pesticide detection and detection of other toxic compounds.

Keywords: graphene, 2D nanomaterials, biosensing, chip design

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Authors: Funmilola O. Omoya

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Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P0.05) from those obtained from the second instars group in all the treatments. Lethal concentration (LC50) at 24 hours for 2nd instars was 2.35 while LC50 at 48 hours was 4.31.This study revealed that Bacillus subtilis possess good larvicidal potential for use in the control of Musca domestica in poultry farms.

Keywords: Bacillus subtilis, Musca domestica, larvicidal activities, poultry farms

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Authors: Irvan Adhin Cholilie, Susinggih Wijana, Yusron Sugiarto

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The aim of this research is to compare the results of black grass jelly produced using laboratory scale and double scale. In this research, the production from the laboratory scale is using ingredients of 1 kg black grass jelly added with 5 liters of water, while the double scale is using 5 kg black grass jelly and 75 liters of water. The results of organoleptic tests performed by 30 panelists (general) to the sample gels of grass black powder produced from both of laboratory and double scale are not different significantly in color, odor, flavor, and texture. Proximate test results conducted in both of grass black jelly powder produced in laboratory scale and double scale also have no significant differences in all parameters. Grass black jelly powder from double scale contains water, carbohydrate, crude fiber, and yield in the amount of 12,25 %; 43,7 %; 5,89 %; and 16,28 % respectively. The results of the energy efficiency analysis by boiling, draining, evaporation, drying, and milling processes are 85,11 %; 76,97 %; 99,64 %; 99,99% and 99,39% respectively. The utility needs including water needs for each batch amounted 0.1 m3 and cost Rp 220,5 per batch, the electricity needs for each batch is 20.01 kWh and cost Rp 18569.28 per batch, and LPG needs for each batch is 30 kg costed Rp 234,000.00 so that the total cost spent for the process is Rp 252,789.78 .

Keywords: black grass jelly, powder, mass balance, energy balance, cost

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Authors: Esmaeil Ahmadinia, Frank Bullen, Ron Ayers

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Many concerns have been raised in recent years about the adequacy of existing creep test methods for evaluating rut-resistance of asphalt mixes. Many researchers believe the main reason for the creep tests being unable to duplicate field results is related to a lack of a realistic confinement for laboratory specimens. In-situ asphalt under axle loads is surrounded by a mass of asphalt, which provides stress-strain generated confinement. However, most existing creep tests are largely unconfined in their nature. It has been hypothesised that by providing a degree of confinement, representative of field conditions, in a creep test, it could be possible to establish a better correlation between the field and laboratory. In this study, a new methodology is explored where confinement for asphalt specimens is provided. The proposed methodology is founded on the current Australian test method, adapted to provide simulated field conditions through the provision of sample confinement.

Keywords: asphalt mixture, creep test, confinements, permanent deformation

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Authors: G. Quiroga-Cubides, A. Díaz, M. Gómez

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The genus Azotobacter has been widely used as bio-fertilizer due to its significant effects on the stimulation and promotion of plant growth in various agricultural species of commercial interest. In order to obtain significantly viable cellular concentration, a scale-up strategy for a liquid fermentation process (SmF) with two strains of A. chroococcum (named Ac1 and Ac10) was validated and adjusted at laboratory and pilot scale. A batch fermentation process under previously defined conditions was carried out on a biorreactor Infors®, model Minifors of 3.5 L, which served as a baseline for this research. For the purpose of increasing process efficiency, the effect of the reduction of stirring speed was evaluated in combination with a fed-batch-type fermentation laboratory scale. To reproduce the efficiency parameters obtained, a scale-up strategy with geometric and fluid dynamic behavior similarities was evaluated. According to the analysis of variance, this scale-up strategy did not have significant effect on cellular concentration and in laboratory and pilot fermentations (Tukey, p > 0.05). Regarding air consumption, fermentation process at pilot scale showed a reduction of 23% versus the baseline. The percentage of reduction related to energy consumption reduction under laboratory and pilot scale conditions was 96.9% compared with baseline.

Keywords: Azotobacter chroococcum, scale-up, liquid fermentation, fed-batch process

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Authors: Jayprakash Yadav, Nivedita Patra

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Polyhydroxybutyrate (PHB) is an interesting material in the field of medical science, pharmaceutical industries, and tissue engineering because of its properties such as biodegradability, biocompatibility, hydrophobicity, and elasticity. PHB is naturally accumulated by several microbes in their cytoplasm during the metabolic process as energy reserve material. PHB can be extracted from cell biomass using halogenated hydrocarbons, chemicals, and enzymes. In this study, a cheaper and non-toxic solvent, acetone, was used for the extraction process. The different parameters like acetone percentage, and solvent pH, process temperature, and incubation periods were optimized using the Response Surface Methodology (RSM). RSM was performed and the determination coefficient (R2) value was found to be 0.8833 from the quadratic regression model with no significant lack of fit. The designed RSM model results indicated that the fitness of the response variable was significant (P-value < 0.0006) and satisfactory to denote the relationship between the responses in terms of PHB recovery and purity with respect to the values of independent variables. Optimum conditions for the maximum PHB recovery and purity were found to be solvent pH 7, extraction temperature - 43 °C, incubation time - 70 minutes, and percentage acetone – 30 % from this study. The maximum predicted PHB recovery was found to be 0.845 g/g biomass dry cell weight and the purity was found to be 97.23 % using the optimized conditions.

Keywords: acetone, PHB, RSM, halogenated hydrocarbons, extraction, bacillus subtilis.

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Authors: Abraham Motlhabane

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This research analysed the written views of science education students regarding the teaching of laboratory work using the online mode. The research adopted the qualitative methodology. The qualitative research was aimed at investigating small and distinct groups normally regarded as a single-site study. Qualitative research was used to describe and analyze the phenomena from the student’s perspective. This means the research began with assumptions of the world view that use theoretical lenses of research problems inquiring into the meaning of individual students. The research was conducted with three groups of students studying for Postgraduate Certificate in Education, Bachelor of Education and honors Bachelor of Education respectively. In each of the study programmes, the science education module is compulsory. Five science education students from each study programme were purposively selected to participate in this research. Therefore, 15 students participated in the research. In order to analysis the data, the data were first printed and hard copies were used in the analysis. The data was read several times and key concepts and ideas were highlighted. Themes and patterns were identified to describe the data. Coding as a process of organising and sorting data was used. The findings of the study are very diverse; some students are in favour of online laboratory whereas other students argue that science can only be learnt through hands-on experimentation.

Keywords: online learning, laboratory work, views, perceptions

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Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im

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Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by zona pellucida with layer of cumulus cells ranging between 59.29-202.14 µm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 µg/mL porcine follicle-stimulating hormone, 1 µg/mL LH, 1µg/mL estradiol with ethanol, and 50 µg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: cumulus cells, electron microscopy, in vitro, porcine oocyte

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Authors: Jer Ping Ooi, Shah Rizal Bin Kasim, Nor Aini Saidin

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The objective of this study was to synthesize nano-sized β-tricalcium phosphate (β-TCP) powder and assess its cytotoxic effects on human osteoblast (hFOB1.19) by using four cytotoxicity assays, namely, lactose dehydrogenase (LDHe), tetrazolium hydroxide (XTT), neutral red (NR), and sulforhodamine B (SRB) assays. β-tricalcium phosphate (β-TCP) is a calcium phosphate compound commonly used as an implant material. To date, bulk-sized β-TCP is reported to be readily tolerated by the osteogenic cells and body based on in vitro, in vivo experiments and clinical studies. However, to what extent of nano-sized β-TCP will react in models as compared to bulk β-TCP is yet to be investigated. Thus, in this project, the cells were treated with nano β-TCP powder within a range of concentrations from 0 to 1000 μg/mL for 24, 48, and 72 h. The cytotoxicity tests showed that loss of cell viability ( > 50%) was high for hFOB1.19 cells in all assays. Cell cycle and apoptosis analysis of hFOB1.19 cells revealed that 50 μg/mL of the compound led to 30.5% of cells being apoptotic after 72 h of incubation, and the percentage was increased to 58.6% when the concentration was increased to 200 μg/mL. When the incubation time was increased from 24 to 72 h, the percentage of apoptotic cells increased from 17.3% to 58.6% when the hFOB1.19 were exposed with 200 μg/mL of nano β-TCP powder. Thus, both concentration and exposure duration affected the cytotoxicity effects of the nano β-TCP powder on hFOB1.19. We hypothesize that these cytotoxic effects on hFOB1.19 are related to the nano-scale size of the β-TCP.

Keywords: β-tricalcium phosphate, hFOB1.19, adipose-derived mesenchymal stem cells, cytotoxicity

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Authors: Nordila Ahmad, Thamer Mohammad, Bruce W. Melville, Zuliziana Suif

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Current methods for predicting local scour at wide bridge piers, were developed on the basis of laboratory studies and very limited scour prediction were tested with field data. Laboratory wide pier scour equation from previous findings with field data were presented. A wide range of field data were used and it consists of both live-bed and clear-water scour. A method for assessing the quality of the data was developed and applied to the data set. Three other wide pier-scour equations from the literature were used to compare the performance of each predictive method. The best-performing scour equation were analyzed using statistical analysis. Comparisons of computed and observed scour depths indicate that the equation from the previous publication produced the smallest discrepancy ratio and RMSE value when compared with the large amount of laboratory and field data.

Keywords: field data, local scour, scour equation, wide piers

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Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme

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Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.

Keywords: ogi, fermentation, bacterial quality, lactic acid bacteria, maize

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Authors: Kashif Ahmed

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Recent years researchers have been motivated towards extensive exploring of living organism, which could be utilized effectively in intense industrial conditions. The present study shows enhanced production, purification and characterization of industrial enzyme, invertase (Beta-D-fructofuranosidase) from Penicillium lilacinum. Various agricultural based by-products (cotton stalk, sunflower waste, rice husk, molasses and date syrup) were used as energy source. The highest amount of enzyme (13.05 Units/mL) was produced when the strain was cultured on growth medium containing date syrup as energy source. Yeast extract was used as nitrogen source after 96 h of incubation at incubation temperature of 40º C. Initial pH of medium was 8.0, inoculum size 6x10⁶ conidia and 200 rev/min agitation rate. The enzyme was also purified (7 folds than crude) and characterized. Molecular mass of purified enzyme (65 kDa) was determined by 10 % SDS-PAGE. Lineweaver-Burk Plot was used to determine Kinetic constants (Vmax 178.6 U/mL/min and Km 2.76 mM). Temperature and pH optima were 55º C and 5.5 respectively. MnCl₂ (52.9 %), MgSO₄ (48.9 %), BaCl₂ (24.6 %), MgCl₂ (9.6 %), CoCl₂ (5.7 %) and NaCl (4.2 %) enhanced the relative activity of enzyme and HgCl₂ (-92.8 %), CuSO₄ (-80.2 %) and CuCl₂ (-76.6 %) were proved inhibitors. The strain was showing enzyme activity even at extreme conditions of temperature (up to 60º C) and pH (up to 9), so it can be used in industries.

Keywords: invertase, Penicillium lilacinum, submerged fermentation, industrial enzyme

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Authors: A. Javorova, J. Oravcova, K. Velisek

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The article summarizes the experience of technical subjects teaching methodologies using a number of software products to solve specific assigned tasks described in this paper. Task is about the problems of automation and mechanization in the industry. Specifically, it focuses on introducing automation in the wood industry. The article describes the design of the automation process for marking wooden hammers. Similar problems are solved by students in CA laboratory.

Keywords: CA system, education, simulation, subject

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Authors: Z. Mmango, U. Nwodo, L. V. Mabinya, A. I. Okoh

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Cellulose and hemicellulose account for a large portion of the world‘s plant biomass. In nature, these polysaccharides are intertwined forming complex materials that requires multiple and expensive treatment processes to free up the raw materials trapped in the matrix. Enzymatic degradation remains as the preferred technique as it is inexpensive and eco-friendly. However, the insufficiencies of enzyme battery systems in the degradation of lignocellulosic complex motivate the search for effective degrading enzymes from bacterial isolates from uncommon environment. The study aimed at the evaluation of actinomycetes isolated from saw dust samples collected from wood factory under bed. Cellulase and xylanase production was screened through organism culture on carboxyl methyl cellulose agar and Birchwood xylan. Halo zone indicating lignocellose utilization was shown by an isolate identified through 16S rRNA gene as Micrococcus luteus. The optimum condition for the production of cellulase and xylanase were incubation temperature of 25 °C, fermentation medium pH 5 and 10, agitation speed of 50 and 200 (rpm) and fermentation incubation time of 96 and 84 (h) respectively. The high cellulose and xylanase activity obtained from this isolate portends industrial relevance.

Keywords: carboxyl methyl cellulose, birchwood xylan, optimization, cellulase, xylanase, micrococcus, DNS method

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Authors: Blerim Olluri

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A forensic science laboratory in Kosovo has never been organized at the level of most modern forensic science laboratories. This was made possible after the war of 1999 with the help and support from the United States. The United States Government/ICITAP provided 9.5 million dollars to support this project, this support have greatly benefitted law enforcement in Kosovo. With the establishment of Operative Procedures of Work and the law for Kosovo Agency of Forensic, the accreditation with ISO/IEC 17025 of the KAF labs it becomes mandatory. Since 2012 Laboratory’s DNA/Serology and Narcotics has begun reviewing and harmonizing their procedures according to ISO/IEC 17025. The focus of this work was to create quality manuals, procedures, work instructions, quality documentation and quality records. Furthermore, during this time is done the validation of work methods from scientific qualified personnel of KAF, without any help from other foreign agencies or accreditation body.In October 2014 we had the first evaluation based on ISO 17025 standards. According to the initial report of this assessment we have non conformity in test and Calibration methods method’s, and accommodation and environmental conditions. We identified several issues that are of extreme importance to KAF. One the most important issue is to create a professional group with experts of KAF, which will work in all the obligations, requested from ISO/IEC 17025. As conclusions that we earn in this path of accreditation, are that laboratory’s need to take corrective action, and all nonconformance’s must be addressed and corrective action taken before accreditation can be granted.

Keywords: accreditation, assessment, narcotics, DNA

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Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

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Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

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Authors: Jolanta Jaskowska, Anna Drabczyk, Sonia Kudlacik, Agnieszka Sobczak-Kupiec, Bozena Tyliszczak

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The aim of the study was to select the best conditions for the synthesis of Beetosan's hydrogels with yellow tea. The study determined recipe hydrogel matrix by selecting the appropriate ratio of substrates and to investigate the effect of yellow tea, on the structure and properties of the hydrogel materials. The scope of the research included both to obtain of raw materials required for the synthesis of hydrogel materials, as well as an assessment of their properties. In the first stage of research Beetosan (chitosan derived from bees), and extract the yellow tea China Kekecha was obtained. The second stage was synthesis hydrogels modified by yellow tea. The synthesis of polymeric matrix was preparation under UV radiation. Obtained hydrogel materials were investigated extensively using incubation investigations, absorption capacity, and spectroscopic (FT-IR) and X-ray diffraction (XRD) methods. Moreover, there was also performed the surface wettability test and a photomicrograph of the structure using scanning electron microscope. Analysis of the obtained results confirms that presence of yellow tea does not significantly affect the behavior of the hydrogels in the incubation fluids. The results show that hydrogel materials exhibit compatibility with the incubatory solutions and they also retain the stability in the tested liquids. Hydrogels obtained in this method might be applied in the cosmetics industry and in the field of medicine. This is possible due to the many interesting properties of tea and biocompatibility and non-toxicity hydrogel materials. The authors would like to thank the The National Centre for Research and Development (Grant no: LIDER/033/697/L-5/13/NCBR/2014) for providing financial support to this project.

Keywords: Beetosan, hygrogels, materials, yellow tea

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Authors: Anja Jutraz, Tadeja Zupancic

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This article explores urban laboratory, which presents a combination of different physical and digital methods and tools for public participation in urban design. The city consists of built and unbuilt environments, which can be defined as a community of people, who live there. Communities should have the option to express opinions and decide about the future of their city, from the early stages of the design process onwards. In this paper, we presented the possibility of involving community into renewal of Banska Štiavnica in Slovakia (more exactly the old mining shaft and lake Michal Štolna) and the methods to promote the community building. As a case study we presented the eTHNo project, Education about Technical, Historical and Natural opportunities of Michal Štolna. Moreover, we discussed the possibility of using virtual digital tools for public participation in urban design, where we especially focused on Virtual Urban Laboratory, VuLab.

Keywords: community building, digital tools, public participation, urban design

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Authors: Khadidja Mebarki, Naceur Boussouar, Nabila Ihaddadene, M. Akermi

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Food products are particularly sensitive, since they concern the health of the consumer, whether it’s be from the health point of view or commercial, this kind of product must be subjected to rigorous controls, in order to prevent any fraud. Quality and safety are essential for food security, public health and economic development. The strengthening of food security is essential to increase food security which is considered reached when all individuals can at any time access safe and nutritious food they need to lead healthy and active lives. The objective of this project is to initiate a quality approach in the laboratories of the quality control and the repression of fraud. It will be directed towards the application of good laboratory practices, traceability, management of quality documents (quality, procedures and specification manual) and quality audits. And to prepare the ground for a possible accreditation by ISO 17025 standard of BECHAR laboratory’s. The project will take place in four main stages: 1- Preparation of an audit grid; 2- Realization of a quality audit according to the method of 5 M completed by a section on quality documentation; 3- Drafting of an audit report and proposal for recommendations; 4- Implementation of corrective actions on the ground. This last step consisted in the formalization of the cleaning disinfection plan; work on good hygiene practices, establishment of a mapping of processes and flow charts of the different processes of the laboratory, classifying quality documents and formalizing the process of document management. During the period of the study within the laboratory, all facets of the work were almost appreciated, as we participated in the expertise performed in within it.

Keywords: quality, management, ISO 17025 accreditation, GLP

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Authors: Yanhua Zhao, Chenli Rao, Dongdong Li, Chuanmin Tao

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The Chinese Centers for Disease Control and Prevention (CCDC) issued a new laboratory-based algorithm for HIV diagnosis on April 2016, which initially screens with a combination HIV-1/HIV-2 antigen/antibody fourth-generation immunoassay (IA) followed, when reactive, an HIV-1/HIV-2 undifferentiated antibody IA in duplicate. Reactive specimens with concordant results undergo supplemental tests with western blots, or HIV-1 nucleic acid tests (NATs) and non-reactive specimens with discordant results receive HIV-1 NATs or p24 antigen tests or 2-4 weeks follow-up tests. However, little data evaluating the application of the new algorithm have been reported to date. The study was to evaluate the performance of new laboratory-based HIV diagnostic algorithm in an inpatient population of Southwest China over the initial 6 months by compared with the old algorithm. Plasma specimens collected from inpatients from May 1, 2016, to October 31, 2016, are submitted to the laboratory for screening HIV infection performed by both the new HIV testing algorithm and the old version. The sensitivity and specificity of the algorithms and the difference of the categorized numbers of plasmas were calculated. Under the new algorithm for HIV diagnosis, 170 of the total 52 749 plasma specimens were confirmed as positively HIV-infected (0.32%). The sensitivity and specificity of the new algorithm were 100% (170/170) and 100% (52 579/52 579), respectively; while 167 HIV-1 positive specimens were identified by the old algorithm with sensitivity 98.24% (167/170) and 100% (52 579/52 579), respectively. Three acute HIV-1 infections (AHIs) and two early HIV-1 infections (EHIs) were identified by the new algorithm; the former was missed by old procedure. Compared with the old version, the new algorithm produced fewer WB-indeterminate results (2 vs. 16, p = 0.001), which led to fewer follow-up tests. Therefore, the new HIV testing algorithm is more sensitive for detecting acute HIV-1 infections with maintaining the ability to verify the established HIV-1 infections and can dramatically decrease the greater number of WB-indeterminate specimens.

Keywords: algorithm, diagnosis, HIV, laboratory

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Authors: Reena K., Mamatha H. G., Somshekarayya, P. Kumar

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Objectives: The annual proficiency testing of intermediate reference laboratories is conducted by the National Reference Laboratory (NRL) to assess the efficiency of the laboratories to correctly identify Mycobacterium tuberculosis and to determine its drug susceptibility pattern. The proficiency testing results from 2013 to 2017 were analyzed to determine laboratories that were consistent in reporting quality results and those that had difficulty in doing so. Methods: A panel of twenty cultures were sent out to each of these laboratories. The laboratories were expected to grow the cultures in their own laboratories, set up drug susceptibly testing by all the methods they were certified for and report the results within the stipulated time period. The turnaround time for reporting results, specificity, sensitivity positive and negative predictive values and efficiency of the laboratory in identifying the cultures were analyzed. Results: Most of the laboratories had reported their results within the stipulated time period. However, there was enormous delay in reporting results from few of the laboratories. This was mainly due to improper functioning of the biosafety level III laboratory. Only 40% of the laboratories had 100% efficiency in solid culture using Lowenstein Jensen medium. This was expected as a solid culture, and drug susceptibility testing is not used for diagnosing drug resistance. Rapid molecular methods such as Line probe assay and Genexpert are used to determine drug resistance. Automated liquid culture system such as the Mycobacterial growth indicator tube is used to determine prognosis of the patient while on treatment. It was observed that 90% of the laboratories had achieved 100% in the liquid culture method. Almost all laboratories had achieved 100% efficiency in the line probe assay method which is the method of choice for determining drug-resistant tuberculosis. Conclusion: Since the liquid culture and line probe assay technologies are routinely used for the detection of drug-resistant tuberculosis the laboratories exhibited higher level of efficiency as compared to solid culture and drug susceptibility testing which are rarely used. The infrastructure of the laboratory should be maintained properly so that samples can be processed safely and results could be declared on time.

Keywords: annual proficiency testing, drug susceptibility testing, intermediate reference laboratory, national reference laboratory

Procedia PDF Downloads 166

Authors: Rebecca Dewfall, Mark Coleman, Vladimir Basabe

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Oxide scale growth is an inevitable byproduct of the high temperature processing of steel. Blister is a phenomenon that occurs due to oxide growth, where high temperatures result in the swelling of surface scale, producing a bubble-like feature. Blisters can subsequently become embedded in the steel substrate during hot rolling in the finishing mill. This rolled in scale defect causes havoc within industry, not only with wear on machinery but loss of customer satisfaction, poor surface finish, loss of material, and profit. Even though blister is a highly prevalent issue, there is still much that is not known or understood. The classic iron oxidation system is a complex multiphase system formed of wustite, magnetite, and hematite, producing multi-layered scales. Each phase will have independent properties such as thermal coefficients, growth rate, and mechanical properties, etc. Furthermore, each additional alloying element will have different affinities for oxygen and different mobilities in the oxide phases so that oxide morphologies are specific to alloy chemistry. Therefore, blister regimes can be unique to each steel grade resulting in a diverse range of formation mechanisms. Laboratory conditions were selected to simulate industrial hot rolling with temperature ranges approximate to the formation of secondary and tertiary scales in the finishing mills. Samples with composition: 0.15Wt% C, 0.1Wt% Si, 0.86Wt% Mn, 0.036Wt% Al, and 0.028Wt% Cr, were oxidised in a thermo-gravimetric analyser (TGA), with an air velocity of 10litresmin-1, at temperaturesof 800°C, 850°C, 900°C, 1000°C, 1100°C, and 1200°C respectively. Samples were held at temperature in an argon atmosphere for 10minutes, then oxidised in air for 600s, 60s, 30s, 15s, and 4s, respectively. Oxide morphology and Blisters were characterised using EBSD, WDX, nanoindentation, FIB, and FEG-SEM imaging. Blister was found to have both a nucleation and growth process. During nucleation, the scale detaches from the substrate and blisters after a very short period, roughly 10s. The steel substrate is then exposed inside of the blister and further oxidised in the reducing atmosphere of the blister, however, the atmosphere within the blister is highly dependent upon the porosity of the blister crown. The blister crown was found to be consistently between 35-40um for all heating regimes, which supports the theory that the blister inflates, and the oxide then subsequently grows underneath. Upon heating, two modes of blistering were identified. In Mode 1 it was ascertained that the stresses produced by oxide growth will increase with increasing oxide thickness. Therefore, in Mode 1 the incubation time for blister formation is shortened by increasing temperature. In Mode 2 increase in temperature will result in oxide with a high ductility and high oxide porosity. The high oxide ductility and/or porosity accommodates for the intrinsic stresses from oxide growth. Thus Mode 2 is the inverse of Mode 1, and incubation time is increased with temperature. A new phenomenon was reported whereby blister formed exclusively through cooling at elevated temperatures above mode 2.

Keywords: FEG-SEM, nucleation, oxide morphology, surface defect

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Authors: Rabih Al-Kaysi

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When quasi-stable solutions of 9-methylanthracene (pi-electron donor, 0.0005 M) and 1,2,4,5-Tetracyanobenzene (pi-electron acceptor, 0.0005 M) in aqueous sodium dodecyl sulfate (SDS, 0.025 M) were gently mixed, uniform-shaped rectangular charge-transfer nanocrystals precipitated out. These red colored charge-transfer (CT) crystals were composed of a 1:1-mole ratio of acceptor/ donor and are highly insoluble in water/SDS solution. The rectangular crystals morphology is semi hollow with symmetrical twin pockets reminiscent of nanocapsules. For a typical crop of nanocapsules, the dimensions are 21 x 6 x 0.5 microns with an approximate hollow volume of 1.5 x 105 nm3. By varying the concentration of aqueous SDS, mixing duration and incubation temperature, we can control the size and volume of the nanocapsules. The initial number of CT seed nanoparticles, formed by mixing the D and A solutions, determined the number and dimensions of the obtained nanocapsules formed after several hours of incubation under still conditions. Prolonged mixing of the donor and acceptor solutions resulted in plenty of initial seeds hence smaller nanocapsules. Short mixing times yields less seed formation and larger micron-sized capsules. The addition of Doxorubicin in situ with the quasi-stable solutions while mixing leads to the formation of CT nanocapsules with Doxorubicin sealed inside. The Doxorubicin can be liberated from the nanocapsules by cracking them using ultrasonication. This method can be extended to other binary CT complex crystals as well.

Keywords: charge-transfer, nanocapsules, nanocrystals, doxorubicin

Procedia PDF Downloads 193

Authors: M. B. Ahmadi, A. A. Kaffash B., B. Mobaraki

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Slump loss, which refers to the gradual reduction of workability and the amount of slump in fresh concrete over time, is one of the significant challenges in the ready-mixed concrete industry. Therefore, having accurate knowledge of the factors affecting slump loss is a crucial solution in this field. In this paper, an attempt was made to investigate the effect of cement produced by different units on the slump of concrete in a laboratory setting. For this purpose, 12 cement samples were prepared from 6 different production units. Physical and chemical tests were performed on the cement samples. Subsequently, a laboratory concrete mix with a slump of 13 ± 1 cm was prepared with each cement sample, and the slump was measured at 0, 15, 30, 45, and 60 minutes. Although the environmental factors, mix design specifications, and execution conditions—factors that significantly influence the slump loss trend—were constant in all 12 laboratory concrete mixes, the slump loss trends differed among them. These trends were categorized based on the results, and the relationship between the slump loss percentage in 60 minutes, the water-cement ratio, and the LOI and K2O values of different cements were introduced.

Keywords: concrete, slump loss, portland cement, efficiency

Procedia PDF Downloads 52

Authors: Manami Masuda, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Fatty acid salts are a type of anionic surfactant and are produced from fatty acids and alkali. Moreover, fatty acid salts are known to have potent antibacterial activities. Acanthamoeba is ubiquitously distributed in the environment including sea water, fresh water, soil and even from the air. Although generally free-living, Acanthamoeba can be an opportunistic pathogen, which could cause a potentially blinding corneal infection known as Acanthamoeba keratitis. So, in this study, we evaluated the anti-amoeba activity of fatty acid salts and fatty acids to Acanthamoeba castellanii ATCC 30010. Materials and Methods: The antibacterial activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), linolenate (C18:3K)) tested on cells of Acanthamoeba castellanii ATCC 30010. Fatty acid salts (concentration of 175 mM and pH 10.5) were prepared by mixing the fatty acid with the appropriate amount of KOH. The amoeba suspension mixed with KOH with a pH adjusted solution was used as the control. Fatty acids (concentration of 175 mM) were prepared by mixing the fatty acid with Tween 80 (20 %). The amoeba suspension mixed with Tween 80 (20 %) was used as the control. The anti-amoeba method, the amoeba suspension (3.0 × 104 cells/ml trophozoites) was mixed with the sample of fatty acid potassium (final concentration of 175 mM). Samples were incubated at 30°C, for 10 min, 60 min, and 180 min and then the viability of A. castellanii was evaluated using plankton counting chamber and trypan blue stainings. The minimum inhibitory concentration (MIC) against Acanthamoeba was determined using the two-fold dilution method. The MIC was defined as the minimal anti-amoeba concentration that inhibited visible amoeba growth following incubation (180 min). Results: C8K, C10K, and C12K were the anti-amoeba effect of 4 log-unit (99.99 % growth suppression of A. castellanii) incubated time for 180 min against A. castellanii at 175mM. After the amoeba, the suspension was mixed with C10K or C12K, destroying the cell membrane had been observed. Whereas, the pH adjusted control solution did not exhibit any effect even after 180 min of incubation with A. castellanii. Moreover, C6, C8, and C18:3 were the anti-amoeba effect of 4 log-unit incubated time for 60 min. C4 and C18:2 exhibited a 4-log reduction after 180 min incubation. Furthermore, the minimum inhibitory concentration (MIC) was determined. The MIC of C10K, C12K and C4 were 2.7 mM. These results indicate that C10K, C12K and C4 have high anti-amoeba activity against A. castellanii and suggest C10K, C12K and C4 have great potential for antimi-amoeba agents.

Keywords: Fatty acid salts, anti-amoeba activities, Acanthamoeba, fatty acids

Procedia PDF Downloads 456

Authors: Meriem Taleb, Ghania Tail, Fatma Zohra Kara, Brahim Djedouani, T. Moussa

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Forensic entomology is the use of insects and their arthropod relatives as silent witnesses to aid legal investigations by interpreting information concerning a death. The main purpose of forensic entomology is to establish the postmortem interval or PMI Postmortem interval is a matter of crucial importance in the investigations of homicide and other untimely deaths when the body found is after three days. Forensic entomology has grown immensely as a discipline in the past thirty years. In Algeria, forensic entomology was introduced in 2010 by the National Institute for Criminalistics and Criminology of the National Gendarmerie (NICC). However, all the work that has been done so far in this growing field in Algeria has been unknown at both the national and international levels. In this context, the aim of this paper is to describe the state of forensic entomology in Algeria. The Laboratory of Entomology of the NICC is the only one of its kind in Algeria. It started its activities in 2010, consisting of two specialists. The main missions of the laboratory are estimation of the PMI by the analysis of entomological evidence, and determination if the body was moved. Currently, the laboratory is performing different tasks such as the expert work required by investigators to estimate the PMI using the insects. The estimation is performed by the accumulated degree days method (ADD) in most of the cases except for those where the cadaver is in dry decay. To assure the quality of the entomological evidence, crime scene personnel are trained by the laboratory of Entomology of the NICC. Recently, undergraduate and graduate students have been studying carrion ecology and insect activity in different geographic locations of Algeria using rabbits and wild boar cadavers as animal models. The Laboratory of Entomology of the NICC has also been involved in some of these research projects. Entomotoxicology experiments are also conducted with the collaboration of the Toxicology Department of the NICC. By dint of hard work that has been performed by the Laboratory of Entomology of the NICC, official bodies have been adopting more and more the use of entomological evidence in criminal investigations in Algeria, which is commendable. It is important, therefore, that steps are taken to fill in the gaps in the knowledge necessary for entomological evidence to have a useful future in criminal investigations in Algeria.

Keywords: forensic entomology, corpse, insects, postmortem interval, expertise, Algeria

Procedia PDF Downloads 381

Authors: Madeline Tagg, Choihoong Mui, Elizabeth Lek, Jide Menakaya

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Introduction: Simulation technology is used by neonatal teams to learn and refresh skills and gain the knowledge and confidence to care for sick neonates. In-situ simulation is considered superior to laboratory-based programmes as it closely mirrors real life situations. This study reports our experience of running regular laboratory-based simulation sessions for senior clinicians and nurses and its impact on their knowledge, skills and confidence. Methods: A before and after questionnaire survey was carried out on senior clinicians and nurses that attended a scheduled laboratory-based simulation session. Participants were asked to document their expectations before a 3-hour monthly laboratory programme started and invited to feedback their reflections at the end of the session. The session included discussion of relevant clinical guidelines, immersion in a scenario and video led debrief. The results of the survey were analysed in three skills based categories - improved, no change or a worsened experience. Results: 45 questionnaires were completed and analysed. Of these 25 (55%) were completed by consultants seven and six by nurses and trainee doctors respectively, and seven respondents were unknown. 40 (88%) rated the session overall and guideline review as good/excellent, 39 respondents (86%) rated the scenario session good/excellent and 40/45 fed back a good/excellent debrief session. 33 (73%) respondents completed the before and after questionnaire. 21/33 (63%) reflected an improved knowledge, skill or confidence in caring for sick new-bon babies, eight respondents reported no change and four fed back a worse experience after the session. Discussion: Most respondents found the laboratory based structured simulation session beneficial for their professional development. They valued equally the whole content of the programme such as guideline review and equipment training as well as the simulation and debrief sessions. Two out three participants stated their knowledge of caring for sick new-born babies had been transformed positively by the session. Sessions where simulation equipment failed or relevant staff were absent contributed to a poor educational experience. Summary: A regular structured laboratory-based simulation programme with a rich content is a credible educational resource for improving the knowledge, skills and confidence of senior clinicians caring for sick new born babies.

Keywords: knowledge, laboratory based, neonates, simulation

Procedia PDF Downloads 103

Authors: Robin Anderson, David Nisbet

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Nitrate, 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) are biologically active chemicals that can accumulate naturally in rangeland grasses forages consumed by grazing cattle, sheep and goats. While toxic to livestock if accumulations and amounts consumed are high enough, particularly in animals having no recent exposure to the forages, these chemicals are known to be potent inhibitors of methane-producing bacteria inhabiting the rumen. Consequently, there is interest in examining their potential use as anti-methanogenic compounds to decrease methane emissions by grazing ruminants. Presently, rumen microbes, collected freshly from a cannulated Holstein cow maintained on 50:50 corn based concentrate:alfalfa diet were mixed (10 mL fluid) in 18 x 150 mm crimp top tubes with 0.5 of high nitrate-containing barley (Hordeum vulgare; containing 272 µmol nitrate per g forage dry matter), and NPA- or NPOH- containing milkvetch forages (Astragalus canadensis and Astragalus miser containing 80 and 174 soluble µmol NPA or NPOH/g forage dry matter respectively). Incubations containing 0.5 g alfalfa (Medicago sativa) were used as controls. Tubes (3 per each respective forage) were capped and incubated anaerobically (using oxygen free carbon dioxide) for 24 h at 39oC after which time amounts of total gas produced were measured via volume displacement and headspace samples were analyzed by gas chromatography to determine concentrations of hydrogen and methane. Fluid samples were analyzed by gas chromatography to measure accumulations of fermentation acids. A completely randomized analysis of variance revealed that the nitrate-containing barley and both the NPA- and the NPOH-containing milkvetches significantly decreased methane production, by > 50%, when compared to methane produced by populations incubated similarly with alfalfa (70.4 ± 3.6 µmol/ml incubation fluid). Accumulations of hydrogen, which are typically increased when methane production is inhibited, by incubations with the nitrate-containing barley and the NPA- and NPOH-containing milkvetches did not differ from accumulations observed in the alfalfa controls (0.09 ± 0.04 µmol/mL incubation fluid). Accumulations of fermentation acids produced in the incubations containing the high-nitrate barley and the NPA- and NPOH-containing milkvetches likewise did not differ from accumulations observed in incubations containing alfalfa (123.5 ± 10.8, 36.0 ± 3.0, 17.1 ± 1.5, 3.5 ± 0.3, 2.3 ± 0.2, 2.2 ± 0.2 µmol/mL incubation fluid for acetate, propionate, butyrate, valerate, isobutyrate, and isovalerate, respectively). This finding indicates the microbial populations did not compensate for the decreased methane production via compensatory changes in production of fermentative acids. Stoichiometric estimation of fermentation balance revealed that > 77% of reducing equivalents generated during fermentation of the forages were recovered in fermentation products and the recoveries did not differ between the alfalfa incubations and those with the high-nitrate barley or the NPA- or NPOH-containing milkvetches. Stoichiometric estimates of amounts of hexose fermented similarly did not differ between the nitrate-, NPA and NPOH-containing incubations and those with the alfalfa, averaging 99.6 ± 37.2 µmol hexose consumed/mL of incubation fluid. These results suggest that forages containing nitrate, NPA or NPOH may be useful to reduce methane emissions of grazing ruminants provided risks of toxicity can be effectively managed.

Keywords: nitrate, nitropropanol, nitropropionic acid, rumen methane emissions

Procedia PDF Downloads 103

Authors: Ahmad Javid Zia, Abdulkerim Ilgun, Suleyman Kamil Akin, Mustafa Altin

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Curing conditions that help concrete, which is one of the most widely used building materials in construction sector, gain strength today is one the important issues. In this study the varying concrete strength depending on water temperature at curing stage is investigated through tests at laboratory. At laboratory the curing conditions has been determined according to both TS EN 12390-2 and regular construction site while performing the experiments on specimens. Five samples have been taken from concrete and cured under five different curing conditions and the compressive strength results of concrete specimens have been compared. One of these five curing conditions has been prepared accordance with TS EN 12390-2, the sample cured at 20 ± 2 ˚C and accepted as reference samples. Two of the remaining sample groups have been cured in 5 ± 2 ˚C and 15 ± 2 ˚C and the other two have been cured outside of the laboratory. One group of the samples which have been cured outside has been watered twice a day and the other group has not been watered at all. The experiments have been carried out on 150x150x150 mm cube samples of C20 (200 kg/cm2) and C25 (250 kg/cm2). 7 and 28 days compressive strength of specimens have been measured and compared.

Keywords: concrete curing, curing conditions, water temperature, concrete compressive strength

Procedia PDF Downloads 352