Search results for: microbial biomass
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1857

Search results for: microbial biomass

87 Numerical Simulation on Two Components Particles Flow in Fluidized Bed

Authors: Wang Heng, Zhong Zhaoping, Guo Feihong, Wang Jia, Wang Xiaoyi

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Flow of gas and particles in fluidized beds is complex and chaotic, which is difficult to measure and analyze by experiments. Some bed materials with bad fluidized performance always fluidize with fluidized medium. The material and the fluidized medium are different in many properties such as density, size and shape. These factors make the dynamic process more complex and the experiment research more limited. Numerical simulation is an efficient way to describe the process of gas-solid flow in fluidized bed. One of the most popular numerical simulation methods is CFD-DEM, i.e., computational fluid dynamics-discrete element method. The shapes of particles are always simplified as sphere in most researches. Although sphere-shaped particles make the calculation of particle uncomplicated, the effects of different shapes are disregarded. However, in practical applications, the two-component systems in fluidized bed also contain sphere particles and non-sphere particles. Therefore, it is needed to study the two component flow of sphere particles and non-sphere particles. In this paper, the flows of mixing were simulated as the flow of molding biomass particles and quartz in fluidized bad. The integrated model was built on an Eulerian–Lagrangian approach which was improved to suit the non-sphere particles. The constructed methods of cylinder-shaped particles were different when it came to different numerical methods. Each cylinder-shaped particle was constructed as an agglomerate of fictitious small particles in CFD part, which means the small fictitious particles gathered but not combined with each other. The diameter of a fictitious particle d_fic and its solid volume fraction inside a cylinder-shaped particle α_fic, which is called the fictitious volume fraction, are introduced to modify the drag coefficient β by introducing the volume fraction of the cylinder-shaped particles α_cld and sphere-shaped particles α_sph. In a computational cell, the void ε, can be expressed as ε=1-〖α_cld α〗_fic-α_sph. The Ergun equation and the Wen and Yu equation were used to calculate β. While in DEM method, cylinder-shaped particles were built by multi-sphere method, in which small sphere element merged with each other. Soft sphere model was using to get the connect force between particles. The total connect force of cylinder-shaped particle was calculated as the sum of the small sphere particles’ forces. The model (size=1×0.15×0.032 mm3) contained 420000 sphere-shaped particles (diameter=0.8 mm, density=1350 kg/m3) and 60 cylinder-shaped particles (diameter=10 mm, length=10 mm, density=2650 kg/m3). Each cylinder-shaped particle was constructed by 2072 small sphere-shaped particles (d=0.8 mm) in CFD mesh and 768 sphere-shaped particles (d=3 mm) in DEM mesh. The length of CFD and DEM cells are 1 mm and 2 mm. Superficial gas velocity was changed in different models as 1.0 m/s, 1.5 m/s, 2.0m/s. The results of simulation were compared with the experimental results. The movements of particles were regularly as fountain. The effect of superficial gas velocity on cylinder-shaped particles was stronger than that of sphere-shaped particles. The result proved this present work provided a effective approach to simulation the flow of two component particles.

Keywords: computational fluid dynamics, discrete element method, fluidized bed, multiphase flow

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86 Temporal Estimation of Hydrodynamic Parameter Variability in Constructed Wetlands

Authors: Mohammad Moezzibadi, Isabelle Charpentier, Adrien Wanko, Robert Mosé

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The calibration of hydrodynamic parameters for subsurface constructed wetlands (CWs) is a sensitive process since highly non-linear equations are involved in unsaturated flow modeling. CW systems are engineered systems designed to favour natural treatment processes involving wetland vegetation, soil, and their microbial flora. Their significant efficiency at reducing the ecological impact of urban runoff has been recently proved in the field. Numerical flow modeling in a vertical variably saturated CW is here carried out by implementing the Richards model by means of a mixed hybrid finite element method (MHFEM), particularly well adapted to the simulation of heterogeneous media, and the van Genuchten-Mualem parametrization. For validation purposes, MHFEM results were compared to those of HYDRUS (a software based on a finite element discretization). As van Genuchten-Mualem soil hydrodynamic parameters depend on water content, their estimation is subject to considerable experimental and numerical studies. In particular, the sensitivity analysis performed with respect to the van Genuchten-Mualem parameters reveals a predominant influence of the shape parameters α, n and the saturated conductivity of the filter on the piezometric heads, during saturation and desaturation. Modeling issues arise when the soil reaches oven-dry conditions. A particular attention should also be brought to boundary condition modeling (surface ponding or evaporation) to be able to tackle different sequences of rainfall-runoff events. For proper parameter identification, large field datasets would be needed. As these are usually not available, notably due to the randomness of the storm events, we thus propose a simple, robust and low-cost numerical method for the inverse modeling of the soil hydrodynamic properties. Among the methods, the variational data assimilation technique introduced by Le Dimet and Talagrand is applied. To that end, a variational data assimilation technique is implemented by applying automatic differentiation (AD) to augment computer codes with derivative computations. Note that very little effort is needed to obtain the differentiated code using the on-line Tapenade AD engine. Field data are collected for a three-layered CW located in Strasbourg (Alsace, France) at the water edge of the urban water stream Ostwaldergraben, during several months. Identification experiments are conducted by comparing measured and computed piezometric head by means of the least square objective function. The temporal variability of hydrodynamic parameter is then assessed and analyzed.

Keywords: automatic differentiation, constructed wetland, inverse method, mixed hybrid FEM, sensitivity analysis

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85 Effects of Culture Conditions on the Adhesion of Yeast Candida spp. and Pichia spp. to Stainless Steel with Different Polishing and Their Control

Authors: Ružica Tomičić, Zorica Tomičić, Peter Raspor

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An abundant growth of unwanted yeasts in food processing plants can lead to problems in quality and safety with significant financial losses. Candida and Pichia are the genera mainly involved in spoilage of products in the food and beverage industry. These contaminating microorganisms can form biofilms on food contact surfaces, being difficult to eradicate, increasing the probability of microbial survival and further dissemination during food processing. It is well known that biofilms are more resistant to antimicrobial agents compared to planktonic cells and this makes them difficult to eliminate. Among the strategies used to overcome resistance to antifungal drugs and preservatives, the use of natural substances such as plant extracts has shown particular promise, and many natural substances have been found to exhibit antifungal properties. This study aimed to investigated the impact of growth medium (Malt Extract broth (MEB) or Yeast Peptone Dextrose (YPD) broth) and temperatures (7°C, 37°C, 43°C for Candida strains and 7°C, 27°C, 32°C for Pichia strains) on the adhesion of Candida spp. and Pichia spp. to stainless steel (AISI 304) discs with different degrees of surface roughness (Ra = 25.20 – 961.9 nm), a material commonly used in the food industry. We also evaluated the antifungal and antiadhesion activity of plant extracts such as Humulus lupulus, Alpinia katsumadai and Evodia rutaecarpa against C. albicans, C glabrata and P. membranifaciens and investigated whether these plant extracts can interfere with biofilm formation. The adhesion was assessed by the crystal violet staining method, while the broth microdilution method CLSI M27-A3 was used to determine the minimum inhibitory concentration (MIC) of plant extracts. Our results indicated that the nutrient content of the medium significantly influenced the amount of adhered cells of the tested yeasts. The growth medium which resulted in a higher adhesion of C. albicans and C. glabrata was MEB, while for C. parapsilosis and C. krusei was YPD. In the case of P. pijperi and P. membranifaciens, YPD broth was more effective in promoting adhesion than MEB. Regarding the effect of temperature, C. albicans strain adhered to stainless steel surfaces in significantly higher level at a temperature of 43°C, while on the other hand C. glabrata, C. parapsilosis and C. krusei showed a different behavior with significantly higher adhesion at 37°C than at 7°C and 43°C. Further, the adherence ability of Pichia strains was highest at 27°C. Based on the MIC values, all plant extracts exerted significant antifungal effects with MIC values ranged from 100 to 400 μg/mL. It was observed that biofilm of C. glabrata were more resistance to plant extracts as compared to C. albicans. However, extracts of A. katsumadai and E. rutaecarpa promoted the growth and development of the preformed biofilm of P. membranifaciens. Thus, the knowledge of how these microorganisms adhere and which factors affect this phenomenon is of great importance in order to avoid their colonization on food contact surfaces.

Keywords: adhesion, Candida spp., Pichia spp., plant extracts

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84 Engineering Packaging for a Sustainable Food Chain

Authors: Ezekiel Olukayode Akintunde

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There is a high level of inadequate methods at all levels of food supply in the global food industry. The inadequacies have led to vast wastages of food. Hence there is a need to curb the wastages that can later affect natural resources, water resources, and energy to avoid negative impacts on the climate and the environment. There is a need to engage multifaceted engineering packaging approaches for a sustainable food chain to ensure active packaging, intelligent packaging, new packaging materials, and a sustainable packaging system. Packaging can be regarded as an indispensable component approach that can be applied to solve major problems of sustainable food consumption globally; this is about controlling the environmental impact of packed food. The creative innovation will ensure that packaged foods are free from food-borne diseases and food chemical pollution. This paper evaluates the key shortcomings that must be addressed by innovative food packaging to ensure a safe, natural environment that will preserve energy and sustain water resources. Certain solutions, including fabricating microbial biodegradable chemical compounds/polymers from agro-food waste remnants, appear a bright path to ensure a strong and innovative waste-based food packaging system. Over the years, depletion in the petroleum reserves has brought about the emergence of biodegradable polymers as a proper replacement for traditional plastics; moreover, the increase in the production of traditional plastics has raised serious concerns about environmental threats. Biodegradable polymers have proven to be biocompatible, which can also be processed for other useful applications. Therefore, this study will showcase a workable guiding framework for designing a sustainable food packaging system that will not constitute a danger to our present society and that will surely preserve natural water resources. Various assessment methods will be deployed at different stages of the packaging design to enhance the package's sustainability. Every decision that will be made must be facilitated with methods that will be engaged per stage to allow for corrective measures throughout the cycle of the design process. Basic performance appraisal of packaging innovations. Food wastage can result in inimical environmental impacts, and ethical practices must be carried out for food loss at home. An examination in West Africa quantified preventable food wastage over the entire food value chain at almost 180kg per person per year. That is preventable food wastage, 35% of which originated at the household level. Many food losses reported, which happened at the harvesting, storage, transportation, and processing stages, are not preventable and are without much environmental impact because such wastage can be used for feeding. Other surveys have shown that 15%-20% of household food losses can be traced to food packaging. Therefore, new innovative packaging systems can lessen the environmental effect of food wastage to extend shelf‐life to lower food loss in the process distribution chain and at the household level.

Keywords: food packaging, biodegradable polymer, intelligent packaging, shelf-life

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83 Impact of Fluoride Contamination on Soil and Water at North 24 Parganas, West Bengal, India

Authors: Rajkumar Ghosh

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Fluoride contamination is a growing concern in various regions across the globe, including North 24 Parganas in West Bengal, India. The presence of excessive fluoride in the environment can have detrimental effects on crops, soil quality, and water resources. This note aims to shed light on the implications of fluoride contamination and its impact on the agricultural sector in North 24 Parganas. The agricultural lands in North 24 Parganas have been significantly affected by fluoride contamination, leading to adverse consequences for crop production. Excessive fluoride uptake by plants can hinder their growth, reduce crop yields, and impact the quality of agricultural produce. Certain crops, such as paddy, vegetables, and fruits, are more susceptible to fluoride toxicity, resulting in stunted growth, leaf discoloration, and reduced nutritional value. Fluoride-contaminated water, often used for irrigation, contributes to the accumulation of fluoride in the soil. Over time, this can lead to soil degradation and reduced fertility. High fluoride levels can alter soil pH, disrupt the availability of essential nutrients, and impair microbial activity critical for nutrient cycling. Consequently, the overall health and productivity of the soil are compromised, making it increasingly challenging for farmers to sustain agricultural practices. Fluoride contamination in North 24 Parganas extends beyond the soil and affects water resources as well. The excess fluoride seeps into groundwater, making it unsafe for consumption. Long-term consumption of fluoride-contaminated water can lead to various health issues, including dental and skeletal fluorosis. These health concerns pose significant risks to the local population, especially those reliant on contaminated water sources for their daily needs. Addressing fluoride contamination requires concerted efforts from various stakeholders, including government authorities, researchers, and farmers. Implementing appropriate water treatment technologies, such as defluoridation units, can help reduce fluoride levels in drinking water sources. Additionally, promoting alternative irrigation methods and crop diversification strategies can aid in mitigating the impact of fluoride on agricultural productivity. Furthermore, creating awareness among farmers about the adverse effects of fluoride contamination and providing access to alternative water sources are crucial steps toward safeguarding the health of the community and sustaining agricultural activities in the region. Fluoride contamination poses significant challenges to crop production, soil health, and water resources in North 24 Parganas, West Bengal. It is imperative to prioritize efforts to address this issue effectively and implement appropriate measures to mitigate fluoride contamination. By adopting sustainable practices and promoting awareness, the community can work towards restoring the agricultural productivity, soil quality and ensuring access to safe drinking water in the region.

Keywords: fluoride contamination, drinking water, toxicity, soil health

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82 Crosslinked Porous 3-Dimensional Cellulose Nanofibers/Gelatin Based Biocomposite Aerogels for Tissue Engineering Application

Authors: Ali Mirtaghavi, Andy Baldwin, Rajendarn Muthuraj, Jack Luo

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Recent advances in biomaterials have led to utilizing biopolymers to develop 3D scaffolds in tissue regeneration. One of the major challenges of designing biomaterials for 3D scaffolds is to mimic the building blocks similar to the extracellular matrix (ECM) of the native tissues. Biopolymer based aerogels obtained by freeze-drying have shown to provide structural similarities to the ECM owing to their 3D format and a highly porous structure with interconnected pores, similar to the ECM. Gelatin (GEL) is known to be a promising biomaterial with inherent regenerative characteristics owing to its chemical similarities to the ECM in native tissue, biocompatibility abundance, cost-effectiveness and accessible functional groups, which makes it facile for chemical modifications with other biomaterials to form biocomposites. Despite such advantages, gelatin offers poor mechanical properties, sensitive enzymatic degradation and high viscosity at room temperature which limits its application and encourages its use to develop biocomposites. Hydrophilic biomass-based cellulose nanofibrous (CNF) has been explored to use as suspension for biocomposite aerogels for the development of 3D porous structures with excellent mechanical properties, biocompatibility and slow enzymatic degradation. In this work, CNF biocomposite aerogels with various ratios of CNF:GEL) (90:10, 70:30 and 50:50) were prepared by freeze-drying technique, and their properties were investigated in terms of physicochemical, mechanical and biological characteristics. Epichlorohydrin (EPH) was used to investigate the effect of chemical crosslinking on the molecular interaction of CNF: GEL, and its effects on physicochemical, mechanical and biological properties of the biocomposite aerogels. Ultimately, chemical crosslinking helped to improve the mechanical resilience of the resulting aerogels. Amongst all the CNF-GEL composites, the crosslinked CNF: GEL (70:30) biocomposite was found to be favourable for cell attachment and viability. It possessed highly porous structure (porosity of ~93%) with pore sizes ranging from 16-110 µm, adequate mechanical properties (compression modulus of ~47 kPa) and optimal biocompatibility both in-vitro and in-vivo, as well as controlled enzymatic biodegradation, high water penetration, which could be considered a suitable option for wound healing application. In-vivo experiments showed improvement on inflammation and foreign giant body cell reaction for the crosslinked CNF: GEL (70:30) compared to the other samples. This could be due to the superior interaction of CNF with gelatin through chemical crosslinking, resulting in more optimal in-vivo improvement. In-vitro cell culture investigation on human dermal fibroblasts showed satisfactory 3D cell attachment over time. Overall, it has been observed that the developed CNF: GEL aerogel can be considered as a potential scaffold for soft tissue regeneration application.

Keywords: 3D scaffolds, aerogels, Biocomposites , tissue engineering

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81 Spatio-Temporal Variation of Gaseous Pollutants and the Contribution of Particulate Matters in Chao Phraya River Basin, Thailand

Authors: Samart Porncharoen, Nisa Pakvilai

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The elevated levels of air pollutants in regional atmospheric environments is a significant problem that affects human health in Thailand, particularly in the Chao Phraya River Basin. Of concern are issues surrounding ambient air pollution such as particulate matter, gaseous pollutants and more specifically concerning air pollution along the river. Therefore, the spatio-temporal study of air pollution in this real environment can gain more accurate air quality data for making formalized environmental policy in river basins. In order to inform such a policy, a study was conducted over a period of January –December, 2015 to continually collect measurements of various pollutants in both urban and regional locations in the Chao Phraya River Basin. This study investigated the air pollutants in many diverse environments along the Chao Phraya River Basin, Thailand in 2015. Multivariate Analysis Techniques such as Principle Component Analysis (PCA) and Path analysis were utilised to classify air pollution in the surveyed location. Measurements were collected in both urban and rural areas to see if significant differences existed between the two locations in terms of air pollution levels. The meteorological parameters of various particulates were collected continually from a Thai pollution control department monitoring station over a period of January –December, 2015. Of interest to this study were the readings of SO2, CO, NOx, O3, and PM10. Results showed a daily arithmetic mean concentration of SO2, CO, NOx, O3, PM10 reading at 3±1 ppb, 0.5± 0.5 ppm, 30±21 ppb, 19±16 ppb, and 40±20 ug/m3 in urban locations (Bangkok). During the same time period, the readings for the same measurements in rural areas, Ayutthaya (were 1±0.5 ppb, 0.1± 0.05 ppm, 25±17 ppb, 30±21 ppb, and 35±10 ug/m3respectively. This show that Bangkok were located in highly polluted environments that are dominated source emitted from vehicles. Further, results were analysed to ascertain if significant seasonal variation existed in the measurements. It was found that levels of both gaseous pollutants and particle matter in dry season were higher than the wet season. More broadly, the results show that levels of pollutants were measured highest in locations along the Chao Phraya. River Basin known to have a large number of vehicles and biomass burning. This correlation suggests that the principle pollutants were from these anthropogenic sources. This study contributes to the body of knowledge surrounding ambient air pollution such as particulate matter, gaseous pollutants and more specifically concerning air pollution along the Chao Phraya River Basin. Further, this study is one of the first to utilise continuous mobile monitoring along a river in order to gain accurate measurements during a data collection period. Overall, the results of this study can be used for making formalized environmental policy in river basins in order to reduce the physical effects on human health.

Keywords: air pollution, Chao Phraya river basin, meteorology, seasonal variation, principal component analysis

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80 Efficacy of DAPG Producing Fluorescent Pseudomonas for Enhancing Nutrient Use Efficacy, Bio-Control of Soil-Borne Diseases and Yield of Groundnut

Authors: Basavaraj Yenagi, P. Nagaraju, C. R. Patil

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Groundnut (Arachis hypohaea L.) is called as “King of oilseeds” and one of the most important food and cash crops in Indian subcontinent. Yield and quality of oil are negatively correlated with poor or imbalanced nutrition and constant exposure to both biotic and abiotic stress factors. Variety of diseases affect groundnut plant, most of them are caused by fungi and lead to severe yield loss. Imbalanced nutrition increases the concerns of environmental deterioration which includes soil fertility. Among different microbial antagonists, Pseudomonas is common member of the plant growth promoting rhizobacteria microflora present in the rhizosphere of groundnut. These are known to produce a beneficial effect on groundnut due to their high metabolic activity leading to the production of enzymes, exopolysaccharides, secondary metabolites, and antibiotics. The ability of pseudomonas lies on their ability to produce antibiotic metabolites such as 2, 4-diacetylphloroglucinol (DAPG). DAPG can inhibit the growth of fungal pathogens namely collar rot and stem rot and also increase the availability of plant nutrients through increased solubilization and uptake of nutrients. Hence, the present study was conducted for three consecutive years (2014 to 2016) in vertisol during the rainy season to assess the efficacy of DAPG producing fluorescent pseudomonas for enhancing nutrient use efficacy, bio-control of soil-borne diseases and yield of groundnut at University of Agricultural Sciences, Dharwad farm. The experiment was laid out in an RCBD with three replications and seven treatments. The mean of three years data revealed that the effect of DAPG-producing producing fluorescent pseudomonas enhanced groundnut yield, uptake of nitrogen and phosphorus and nutrient use efficiency and also found to be effective in bio-control of collar rot and stem rot incidence leading to increase pod yield of groundnut. Higher dry pod yield of groundnut was obtained with DAPG 2(3535 kg ha-1) closely followed by DAPG 4(3492 kg ha-1), FP 98(3443 kg ha-1), DAPG 1(3414 kg ha-1), FP 86(3361 kg ha-1) and Trichoderma spp. (3380 kg ha-1) over control(3173 kg ha-1). A similar trend was obtained with other growth and yield attributing parameters. N uptake ranged from 8.21 percent to FP 86 to 17.91 percent with DAPG 2 and P uptake ranged between 5.56 percent with FP 86 to 16.67 percent with DAPG 2 over control. The first year, there was no incidence of collar rot. During the second year, the control plot recorded 2.51 percent incidence and it ranged from 0.82 percent to 1.43 percent in different DAPG-producing fluorescent pseudomonas treatments. The similar trend was noticed in the third year with lower incidence. The stem rot incidence was recorded during all the three years. Mean data indicated that the control plot recorded 2.65 percent incidence and it ranged from 0.71 percent to 1.23 percent in different DAPG-producing fluorescent pseudomonas treatments. The increase in net monetary benefits ranged from Rs.5975 ha-1 to Rs.11407 ha 1 in different treatments. Hence, as a low-cost technology, seed treatment with available DAPG-producing fluorescent pseudomonas has a beneficial effect on groundnut for enhancing groundnut yield, nutrient use efficiency and bio-control of soil-borne diseases.

Keywords: groundnut, DAPG, fluorescent pseudomonas, nutrient use efficiency, collar rot, stem rot

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79 Selection and Preparation of High Performance, Natural and Cost-Effective Hydrogel as a Bio-Ink for 3D Bio-Printing and Organ on Chip Applications

Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

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78 LaeA/1-Velvet Interplay in Aspergillus and Trichoderma: Regulation of Secondary Metabolites and Cellulases

Authors: Razieh Karimi Aghcheh, Christian Kubicek, Joseph Strauss, Gerhard Braus

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Filamentous fungi are of considerable economic and social significance for human health, nutrition and in white biotechnology. These organisms are dominant producers of a range of primary metabolites such as citric acid, microbial lipids (biodiesel) and higher unsaturated fatty acids (HUFAs). In particular, they produce also important but structurally complex secondary metabolites with enormous therapeutic applications in pharmaceutical industry, for example: cephalosporin, penicillin, taxol, zeranol and ergot alkaloids. Several fungal secondary metabolites, which are significantly relevant to human health do not only include antibiotics, but also e.g. lovastatin, a well-known antihypercholesterolemic agent produced by Aspergillus. terreus, or aflatoxin, a carcinogen produced by A. flavus. In addition to their roles for human health and agriculture, some fungi are industrially and commercially important: Species of the ascomycete genus Hypocrea spp. (teleomorph of Trichoderma) have been demonstrated as efficient producer of highly active cellulolytic enzymes. This trait makes them effective in disrupting and depolymerization of lignocellulosic materials and thus applicable tools in number of biotechnological areas as diverse as clothes-washing detergent, animal feed, and pulp and fuel productions. Fungal LaeA/LAE1 (Loss of aflR Expression A) homologs their gene products act at the interphase between secondary metabolisms, cellulase production and development. Lack of the corresponding genes results in significant physiological changes including loss of secondary metabolite and lignocellulose degrading enzymes production. At the molecular level, the encoded proteins are presumably methyltransferases or demethylases which act directly or indirectly at heterochromatin and interact with velvet domain proteins. Velvet proteins bind to DNA and affect expression of secondary metabolites (SMs) genes and cellulases. The dynamic interplay between LaeA/LAE1, velvet proteins and additional interaction partners is the key for an understanding of the coordination of metabolic and morphological functions of fungi and is required for a biotechnological control of the formation of desired bioactive products. Aspergilli and Trichoderma represent different biotechnologically significant species with significant differences in the LaeA/LAE1-Velvet protein machinery and their target proteins. We, therefore, performed a comparative study of the interaction partners of this machinery and the dynamics of the various protein-protein interactions using our robust proteomic and mass spectrometry techniques. This enhances our knowledge about the fungal coordination of secondary metabolism, cellulase production and development and thereby will certainly improve recombinant fungal strain construction for the production of industrial secondary metabolite or lignocellulose hydrolytic enzymes.

Keywords: cellulases, LaeA/1, proteomics, secondary metabolites

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77 Inverted Diameter-Limit Thinning: A Promising Alternative for Mixed Populus tremuloides Stands Management

Authors: Ablo Paul Igor Hounzandji, Benoit Lafleur, Annie DesRochers

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Introduction: Populus tremuloides [Michx] regenerates rapidly and abundantly by root suckering after harvest, creating stands with interconnected stems. Pre-commercial thinning can be used to concentrate growth on fewer stems to reach merchantability faster than un-thinned stands. However, conventional thinning methods are typically designed to reach even spacing between residual stems (1,100 stem ha⁻¹, evenly distributed), which can lead to treated stands consisting of weaker/smaller stems compared to the original stands. Considering the nature of P. tremuloides's regeneration, with large underground biomass of interconnected roots, aiming to keep the most vigorous and largest stems, regardless of their spatial distribution, inverted diameter-limit thinning could be more beneficial to post-thinning stand productivity because it would reduce the imbalance between roots and leaf area caused by thinning. Aims: This study aimed to compare stand and stem productivity of P. tremuloides stands thinned with a conventional thinning treatment (CT; 1,100 stem ha⁻¹, evenly distributed), two levels of inverted diameter-limit thinning (DL1 and DL2, keeping the largest 1100 or 2200 stems ha⁻¹, respectively, regardless of their spatial distribution) and a control unthinned treatment. Because DL treatments can create substantial or frequent gaps in the thinned stands, we also aimed to evaluate the potential of this treatment to recreate mixed conifer-broadleaf stands by fill-planting Picea glauca seedlings. Methods: Three replicate 21 year-old sucker-regenerated aspen stands were thinned in 2010 according to four treatments: CT, DL1, DL2, and un-thinned control. Picea glauca seedlings were underplanted in gaps created by the DL1 and DL2 treatments. Stand productivity per hectare, stem quality (diameter and height, volume stem⁻¹) and survival and height growth of fill-planted P. glauca seedlings were measured 8 year post-treatments. Results: Productivity, volume, diameter, and height were better in the treated stands (CT, DL1, and DL2) than in the un-thinned control. Productivity of CT and DL1 stands was similar 4.8 m³ ha⁻¹ year⁻¹. At the tree level, diameter and height of the trees in the DL1 treatment were 5% greater than those in the CT treatment. The average volume of trees in the DL1 treatment was 11% higher than the CT treatment. Survival after 8 years of fill planted P. glauca seedlings was 2% greater in the DL1 than in the DL2 treatment. DL1 treatment also produced taller seedlings (+20 cm). Discussion: Results showed that DL treatments were effective in producing post-thinned stands with larger stems without affecting stand productivity. In addition, we showed that these treatments were suitable to introduce slower growing conifer seedlings such as Picea glauca in order to re-create or maintain mixed stands despite the aggressive nature of P. tremuloides sucker regeneration.

Keywords: Aspen, inverted diameter-limit, mixed forest, populus tremuloides, silviculture, thinning

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76 Treatment with Triton-X 100: An Enhancement Approach for Cardboard Bioprocessing

Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

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75 Development of DNDC Modelling Method for Evaluation of Carbon Dioxide Emission from Arable Soils in European Russia

Authors: Olga Sukhoveeva

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Carbon dioxide (CO2) is the main component of carbon biogeochemical cycle and one of the most important greenhouse gases (GHG). Agriculture, particularly arable soils, are one the largest sources of GHG emission for the atmosphere including CO2.Models may be used for estimation of GHG emission from agriculture if they can be adapted for different countries conditions. The only model used in officially at national level in United Kingdom and China for this purpose is DNDC (DeNitrification-DeComposition). In our research, the model DNDC is offered for estimation of GHG emission from arable soils in Russia. The aim of our research was to create the method of DNDC using for evaluation of CO2 emission in Russia based on official statistical information. The target territory was European part of Russia where many field experiments are located. At the first step of research the database on climate, soil and cropping characteristics for the target region from governmental, statistical, and literature sources were created. All-Russia Research Institute of Hydrometeorological Information – World Data Centre provides open daily data about average meteorological and climatic conditions. It must be calculated spatial average values of maximum and minimum air temperature and precipitation over the region. Spatial average values of soil characteristics (soil texture, bulk density, pH, soil organic carbon content) can be determined on the base of Union state register of soil recourses of Russia. Cropping technologies are published by agricultural research institutes and departments. We offer to define cropping system parameters (annual information about crop yields, amount and types of fertilizers and manure) on the base of the Federal State Statistics Service data. Content of carbon in plant biomass may be calculated via formulas developed and published by Ministry of Natural Resources and Environment of the Russian Federation. At the second step CO2 emission from soil in this region were calculated by DNDC. Modelling data were compared with empirical and literature data and good results were obtained, modelled values were equivalent to the measured ones. It was revealed that the DNDC model may be used to evaluate and forecast the CO2 emission from arable soils in Russia based on the official statistical information. Also, it can be used for creation of the program for decreasing GHG emission from arable soils to the atmosphere. Financial Support: fundamental scientific researching theme 0148-2014-0005 No 01201352499 ‘Solution of fundamental problems of analysis and forecast of Earth climatic system condition’ for 2014-2020; fundamental research program of Presidium of RAS No 51 ‘Climate change: causes, risks, consequences, problems of adaptation and regulation’ for 2018-2020.

Keywords: arable soils, carbon dioxide emission, DNDC model, European Russia

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74 Ruminal Fermentation of Biologically Active Nitrate- and Nitro-Containing Forages

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

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73 COVID-19: Potential Effects of Nutritional Factors on Inflammation Relief

Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

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72 Gene Expression Profiling of Iron-Related Genes of Pasteurella multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar

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Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.

Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes

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71 Hydrogen Production from Auto-Thermal Reforming of Ethanol Catalyzed by Tri-Metallic Catalyst

Authors: Patrizia Frontera, Anastasia Macario, Sebastiano Candamano, Fortunato Crea, Pierluigi Antonucci

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The increasing of the world energy demand makes today biomass an attractive energy source, based on the minimizing of CO2 emission and on the global warming reduction purposes. Recently, COP-21, the international meeting on global climate change, defined the roadmap for sustainable worldwide development, based on low-carbon containing fuel. Hydrogen is an energy vector able to substitute the conventional fuels from petroleum. Ethanol for hydrogen production represents a valid alternative to the fossil sources due to its low toxicity, low production costs, high biodegradability, high H2 content and renewability. Ethanol conversion to generate hydrogen by a combination of partial oxidation and steam reforming reactions is generally called auto-thermal reforming (ATR). The ATR process is advantageous due to the low energy requirements and to the reduced carbonaceous deposits formation. Catalyst plays a pivotal role in the ATR process, especially towards the process selectivity and the carbonaceous deposits formation. Bimetallic or trimetallic catalysts, as well as catalysts with doped-promoters supports, may exhibit high activity, selectivity and deactivation resistance with respect to the corresponding monometallic ones. In this work, NiMoCo/GDC, NiMoCu/GDC and NiMoRe/GDC (where GDC is Gadolinia Doped Ceria support and the metal composition is 60:30:10 for all catalyst) have been prepared by impregnation method. The support, Gadolinia 0.2 Doped Ceria 0.8, was impregnated by metal precursors solubilized in aqueous ethanol solution (50%) at room temperature for 6 hours. After this, the catalysts were dried at 100°C for 8 hours and, subsequently, calcined at 600°C in order to have the metal oxides. Finally, active catalysts were obtained by reduction procedure (H2 atmosphere at 500°C for 6 hours). All sample were characterized by different analytical techniques (XRD, SEM-EDX, XPS, CHNS, H2-TPR and Raman Spectorscopy). Catalytic experiments (auto-thermal reforming of ethanol) were carried out in the temperature range 500-800°C under atmospheric pressure, using a continuous fixed-bed microreactor. Effluent gases from the reactor were analyzed by two Varian CP4900 chromarographs with a TCD detector. The analytical investigation focused on the preventing of the coke deposition, the metals sintering effect and the sulfur poisoning. Hydrogen productivity, ethanol conversion and products distribution were measured and analyzed. At 600°C, all tri-metallic catalysts show the best performance: H2 + CO reaching almost the 77 vol.% in the final gases. While NiMoCo/GDC catalyst shows the best selectivity to hydrogen whit respect to the other tri-metallic catalysts (41 vol.% at 600°C). On the other hand, NiMoCu/GDC and NiMoRe/GDC demonstrated high sulfur poisoning resistance (up to 200 cc/min) with respect to the NiMoCo/GDC catalyst. The correlation among catalytic results and surface properties of the catalysts will be discussed.

Keywords: catalysts, ceria, ethanol, gadolinia, hydrogen, Nickel

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70 Exploring Bio-Inspired Catecholamine Chemistry to Design Durable Anti-Fungal Wound Dressings

Authors: Chetna Dhand, Venkatesh Mayandi, Silvia Marrero Diaz, Roger W. Beuerman, Seeram Ramakrishna, Rajamani Lakshminarayanan

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Sturdy Insect Cuticle Sclerotization, Incredible Substrate independent Mussel’s bioadhesion, Tanning of Leather are some of catechol(amine)s mediated natural processes. Chemical contemplation spots toward a mechanism instigated with the formation of the quinone moieties from the respective catechol(amine)s, via oxidation, followed by the nucleophilic addition of the amino acids/proteins/peptides to this quinone leads to the development of highly strong, cross-linked and water-resistant proteinacious structures. Inspired with this remarkable catechol(amine)s chemistry towards amino acids/proteins/peptides, we attempted to design highly stable and water-resistant antifungal wound dressing mats with exceptional durability using collagen (protein), dopamine (catecholamine) and antifungal drugs (Amphotericin B and Caspofungin) as the key materials. Electrospinning technique has been used to fabricate desired nanofibrous mat including Collagen (COLL), COLL/Dopamine (COLL/DP) and calcium incorporated COLL/DP (COLL-DP-Ca2+). The prepared protein-based scaffolds have been studied for their microscopic investigations (SEM, TEM, and AFM), structural analysis (FT-IR), mechanical properties, water wettability characteristics and aqueous stability. Biocompatibility of these scaffolds has been analyzed for dermal fibroblast cells using MTS assay, Cell TrackerTM Green CMFDA and confocal imaging. Being the winner sample, COLL-DP-Ca2+ scaffold has been selected for incorporating two antifungal drugs namely Caspofungin (Peptide based) and Amphotericin B (Non-Peptide based). Antifungal efficiency of the designed mats has been evaluated for eight diverse fungal strains employing different microbial assays including disc diffusion, cell-viability assay, time kill kinetics etc. To confirm the durability of these mats, in term of their antifungal activity, drug leaching studies has been performed and monitored using disc diffusion assay each day. Ex-vivo fungal infection model has also been developed and utilized to validate the antifungal efficacy of the designed wound dressings. Results clearly reveal dopamine mediated crosslinking within COLL-antifungal scaffolds that leads to the generation of highly stable, mechanical tough, biocompatible wound dressings having the zone of inhabitation of ≥ 2 cm for almost all the investigated fungal strains. Leaching studies and Ex-vivo model has confirmed the durability of these wound dressing for more than 3 weeks and certified their suitability for commercialization. A model has also been proposed to enlighten the chemical mechanism involved for the development of these antifungal wound dressings with exceptional robustness.

Keywords: catecholamine chemistry, electrospinning technique, antifungals, wound dressings, collagen

Procedia PDF Downloads 347
69 Evaluation of Herbal Extracts for Their Potential Application as Skin Prebiotics

Authors: Anja I. Petrov, Milica B. Veljković, Marija M. Ćorović, Ana D. Milivojević, Milica B. Simović, Katarina M. Banjanac, Dejan I. Bezbradica

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One of the fundamental requirements for overall human well-being is a stable and balanced microbiome. Aside from the microorganisms that reside within the body, a large number of microorganisms, especially bacteria, swarming the human skin is in homeostasis with the host and represents a skin microbiota. Even though the immune system of the skin is capable of distinguishing between commensal and potentially harmful transient bacteria, the cutaneous microbial balance can be disrupted under certain circumstances. In that case, a reduction in the skin microbiota diversity, as well as changes in metabolic activity, results in dermal infections and inflammation. Probiotics and prebiotics have the potential to play a significant role in the treatment of these skin disorders. The most common resident bacteria found on the skin, Staphylococcus epidermidis, can act as a potential skin probiotic, contributing to the protection of healthy skin from pathogen colonization, such as Staphylococcus aureus, which is related to atopic dermatitis exacerbation. However, as it is difficult to meet regulations in cosmetic products, another therapy approach could be topical prebiotic supplementation of the skin microbiota. In recent research, polyphenols are attracting scientists' interest as biomolecules with possible prebiotic effects on the skin microbiota. This research aimed to determine how herbal extracts rich in different polyphenolic compounds (lemon balm, St. John's wort, coltsfoot, pine needle, and yarrow) affected the growth of S. epidermidis and S. aureus. The first part of the study involved screening plants to determine if they could be regarded as probable candidates to be skin prebiotics. The effect of each plant on bacterial growth was examined by supplementing the nutrient medium with their extracts and comparing it with control samples (without extract). The results obtained after 24 h of incubation showed that all tested extracts influenced the growth of the examined bacteria to some extent. Since lemon balm and St. John's wort extracts displayed bactericidal activity against S. epidermidis, whereas coltsfoot inhibited both bacteria equally, they were not explored further. On the other hand, pine needles and yarrow extract led to an increase in S. epidermidis/S. aureus ratio, making them prospective candidates to be used as skin prebiotics. By examining the prebiotic effect of two extracts at different concentrations, it was revealed that, in the case of yarrow, 0.1% of extract dry matter in the fermentation medium was optimal, while for the pine needle extract, a concentration of 0.05% was preferred, since it selectively stimulated S. epidermidis growth and inhibited S. aureus proliferation. Additionally, the total polyphenols and flavonoid content of the two extracts were determined, revealing different concentrations and polyphenol profiles. Since yarrow and pine extracts affected the growth of skin bacteria in a dose-dependent manner, by carefully selecting the quantities of these extracts, and thus polyphenols content, it is possible to achieve desirable alterations of skin microbiota composition, which may be suitable for the treatment of atopic dermatitis.

Keywords: herbal extracts, polyphenols, skin microbiota, skin prebiotics

Procedia PDF Downloads 146
68 Exploring Fluoroquinolone-Resistance Dynamics Using a Distinct in Vitro Fermentation Chicken Caeca Model

Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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67 Safety Considerations of Furanics for Sustainable Applications in Advanced Biorefineries

Authors: Anitha Muralidhara, Victor Engelen, Christophe Len, Pascal Pandard, Guy Marlair

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Production of bio-based chemicals and materials from lignocellulosic biomass is gaining tremendous importance in advanced bio-refineries while aiming towards progressive replacement of petroleum based chemicals in transportation fuels and commodity polymers. One such attempt has resulted in the production of key furan derivatives (FD) such as furfural, HMF, MMF etc., via acid catalyzed dehydration (ACD) of C6 and C5 sugars, which are further converted into key chemicals or intermediates (such as Furandicarboxylic acid, Furfuryl alcohol etc.,). In subsequent processes, many high potential FD are produced, that can be converted into high added value polymers or high energy density biofuels. During ACD, an unavoidable polyfuranic byproduct is generated which is called humins. The family of FD is very large with varying chemical structures and diverse physicochemical properties. Accordingly, the associated risk profiles may largely vary. Hazardous Material (Haz-mat) classification systems such as GHS (CLP in the EU) and the UN TDG Model Regulations for transport of dangerous goods are one of the preliminary requirements for all chemicals for their appropriate classification, labelling, packaging, safe storage, and transportation. Considering the growing application routes of FD, it becomes important to notice the limited access to safety related information (safety data sheets available only for famous compounds such as HMF, furfural etc.,) in these internationally recognized haz-mat classification systems. However, these classifications do not necessarily provide information about the extent of risk involved when the chemical is used in any specific application. Factors such as thermal stability, speed of combustion, chemical incompatibilities, etc., can equally influence the safety profile of a compound, that are clearly out of the scope of any haz-mat classification system. Irrespective of the bio-based origin, FD has so far received inconsistent remarks concerning their toxicity profiles. With such inconsistencies, there is a fear that, a large family of FD may also follow extreme judgmental scenarios like ionic liquids, by ranking some compounds as extremely thermally stable, non-flammable, etc., Unless clarified, these messages could lead to misleading judgements while ranking the chemical based on its hazard rating. Safety is a key aspect in any sustainable biorefinery operation/facility, which is often underscored or neglected. To fill up these existing data gaps and to address ambiguities and discrepancies, the current study focuses on giving preliminary insights on safety assessment of FD and their potential targeted by-products. With the available information in the literature and obtained experimental results, physicochemical safety, environmental safety as well as (a scenario based) fire safety profiles of key FD, as well as side streams such as humins and levulinic acid, will be considered. With this, the study focuses on defining patterns and trends that gives coherent safety related information for existing and newly synthesized FD in the market for better functionality and sustainable applications.

Keywords: furanics, humins, safety, thermal and fire hazard, toxicity

Procedia PDF Downloads 149
66 The Product Innovation Using Nutraceutical Delivery System on Improving Growth Performance of Broiler

Authors: Kitti Supchukun, Kris Angkanaporn, Teerapong Yata

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The product innovation using a nutraceutical delivery system on improving the growth performance of broilers is the product planning and development to solve the antibiotics banning policy incurred in the local and global livestock production system. Restricting the use of antibiotics can reduce the quality of chicken meat and increase pathogenic bacterial contamination. Although other alternatives were used to replace antibiotics, the efficacy was inconsistent, reflecting on low chicken growth performance and contaminated products. The product innovation aims to effectively deliver the selected active ingredients into the body. This product is tested on the pharmaceutical lab scale and on the farm-scale for market feasibility in order to create product innovation using the nutraceutical delivery system model. The model establishes the product standardization and traceable quality control process for farmers. The study is performed using mixed methods. Starting with a qualitative method to find the farmers' (consumers) demands and the product standard, then the researcher used the quantitative research method to develop and conclude the findings regarding the acceptance of the technology and product performance. The survey has been sent to different organizations by random sampling among the entrepreneur’s population including integrated broiler farm, broiler farm, and other related organizations. The mixed-method results, both qualitative and quantitative, verify the user and lead users' demands since they provide information about the industry standard, technology preference, developing the right product according to the market, and solutions for the industry problems. The product innovation selected nutraceutical ingredients that can solve the following problems in livestock; bactericidal, anti-inflammation, gut health, antioxidant. The combinations of the selected nutraceutical and nanostructured lipid carriers (NLC) technology aim to improve chemical and pharmaceutical components by changing the structure of active ingredients into nanoparticle, which will be released in the targeted location with accurate concentration. The active ingredients in nanoparticle form are more stable, elicit antibacterial activity against pathogenic Salmonella spp and E.coli, balance gut health, have antioxidant and anti-inflammation activity. The experiment results have proven that the nutraceuticals have an antioxidant and antibacterial activity which also increases the average daily gain (ADG), reduces feed conversion ratio (FCR). The results also show a significant impact on the higher European Performance Index that can increase the farmers' profit when exporting. The product innovation will be tested in technology acceptance management methods from farmers and industry. The production of broiler and commercialization analyses are useful to reduce the importation of animal supplements. Most importantly, product innovation is protected by intellectual property.

Keywords: nutraceutical, nano structure lipid carrier, anti-microbial drug resistance, broiler, Salmonella

Procedia PDF Downloads 146
65 Assessment of Environmental Mercury Contamination from an Old Mercury Processing Plant 'Thor Chemicals' in Cato Ridge, KwaZulu-Natal, South Africa

Authors: Yohana Fessehazion

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Mercury is a prominent example of a heavy metal contaminant in the environment, and it has been extensively investigated for its potential health risk in humans and other organisms. In South Africa, massive mercury contamination happened in1980s when the England-based mercury reclamation processing plant relocated to Cato Ridge, KwaZulu-Natal Province, and discharged mercury waste into the Mngceweni River. This mercury waste discharge resulted in high mercury concentration that exceeded the acceptable levels in Mngceweni River, Umgeni River, and human hair of the nearby villagers. This environmental issue raised the alarm, and over the years, several environmental assessments were reported the dire environmental crises resulting from the Thor Chemicals (now known as Metallica Chemicals) and urged the immediate removal of the around 3,000 tons of mercury waste stored in the factory storage facility over two decades. Recently theft of some containers with the toxic substance from the Thor Chemicals warehouse and the subsequent fire that ravaged the facility furtherly put the factory on the spot escalating the urgency of left behind deadly mercury waste removal. This project aims to investigate the mercury contamination leaking from an old Thor Chemicals mercury processing plant. The focus will be on sediments, water, terrestrial plants, and aquatic weeds such as the prominent water hyacinth weeds in the nearby water systems of Mngceweni River, Umgeni River, and Inanda Dam as a bio-indicator and phytoremediator for mercury pollution. Samples will be collected in spring around October when the condition is favourable for microbial activity to methylate mercury incorporated in sediments and blooming season for some aquatic weeds, particularly water hyacinth. Samples of soil, sediment, water, terrestrial plant, and aquatic weed will be collected per sample site from the point of source (Thor Chemicals), Mngceweni River, Umgeni River, and the Inanda Dam. One-way analysis of variance (ANOVA) tests will be conducted to determine any significant differences in the Hg concentration among all sampling sites, followed by Least Significant Difference post hoc test to determine if mercury contamination varies with the gradient distance from the source point of pollution. The flow injection atomic spectrometry (FIAS) analysis will also be used to compare the mercury sequestration between the different plant tissues (roots and stems). The principal component analysis is also envisaged for use to determine the relationship between the source of mercury pollution and any of the sampling points (Umgeni and Mngceweni Rivers and the Inanda Dam). All the Hg values will be expressed in µg/L or µg/g in order to compare the result with the previous studies and regulatory standards. Sediments are expected to have relatively higher levels of Hg compared to the soils, and aquatic macrophytes, water hyacinth weeds are expected to accumulate a higher concentration of mercury than terrestrial plants and crops.

Keywords: mercury, phytoremediation, Thor chemicals, water hyacinth

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64 Evaluation of Feasibility of Ecological Sanitation in Central Nepal

Authors: K. C. Sharda

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Introduction: In the world, almost half of the population are lacking proper access to improved sanitation services. In Nepal, large number of people are living without access to any sanitation facility. Ecological sanitation toilet which is defined as water conserving and nutrient recycling system for use of human urine and excreta in agriculture would count a lot to utilize locally available resources, to regenerate soil fertility, to save national currency and to achieve the goal of elimination open defecation in country like Nepal. The objectives of the research were to test the efficacy of human urine for improving crop performance and to evaluate the feasibility of ecological sanitation in rural area of Central Nepal. Materials and Methods: The field investigation was carried out at Palung Village Development Committee (VDC) of Makawanpur District, Nepal from March – August, 2016. Five eco-san toilets in two villages (Angare and Bhot Khoriya) were constructed and questionnaire survey was carried out. During the questionnaire survey, respondents were asked about socio-economic parameters, farming practices, awareness of ecological sanitation and fertilizer value of human urine and excreta in agriculture. In prior to a field experiment, soil was sampled for analysis of basic characteristics. In the field experiment, cauliflower was cultivated for a month in the two sites to compare the fertilizer value of urine with chemical fertilizer and no fertilizer with three replications. The harvested plant samples were analyzed to understand the nutrient content in plant with different treatments. Results and Discussion: Eighty three percent respondents were engaged in agriculture growing mainly vegetables, which may raise the feasibility of ecological sanitation. In the study area, water deficiencies in dry season, high demand of chemical fertilizer, lack of sanitation awareness were found to be solved. The soil at Angare has sandier texture and lower nitrogen content compared to that in Bhot Khoriya. While the field experiment in Angare showed that the aboveground biomass of cauliflower in the urine fertilized plot were similar with that in the chemically fertilized plot and higher than those in the non-fertilized plots, no significant difference among the treatments were found in Bhot Khoriya. The more distinctive response of crop growth to the three treatments in the former might be attributed to the poorer soil productivity, which in turn could be caused by the poorer inherent soil fertility and the poorer past management by the farmer in Angare. Thus, use of urine as fertilizer could help poor farmers with low quality soil. The significantly different content of nitrogen and potassium in the plant samples among three treatments in Bhot Khoriya would require further investigation. When urine is utilized as a fertilizer, the productivity could be increased and the money to buy chemical fertilizer would be utilized in other livelihood activities. Ecological sanitation is feasible in the area with similar socio-economic parameter.

Keywords: cauliflower, chemical fertilizer, ecological sanitation, Nepal, urine

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63 Review of Carbon Materials: Application in Alternative Energy Sources and Catalysis

Authors: Marita Pigłowska, Beata Kurc, Maciej Galiński

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The application of carbon materials in the branches of the electrochemical industry shows an increasing tendency each year due to the many interesting properties they possess. These are, among others, a well-developed specific surface, porosity, high sorption capacity, good adsorption properties, low bulk density, electrical conductivity and chemical resistance. All these properties allow for their effective use, among others in supercapacitors, which can store electric charges of the order of 100 F due to carbon electrodes constituting the capacitor plates. Coals (including expanded graphite, carbon black, graphite carbon fibers, activated carbon) are commonly used in electrochemical methods of removing oil derivatives from water after tanker disasters, e.g. phenols and their derivatives by their electrochemical anodic oxidation. Phenol can occupy practically the entire surface of carbon material and leave the water clean of hydrophobic impurities. Regeneration of such electrodes is also not complicated, it is carried out by electrochemical methods consisting in unblocking the pores and reducing resistances, and thus their reactivation for subsequent adsorption processes. Graphite is commonly used as an anode material in lithium-ion cells, while due to the limited capacity it offers (372 mAh g-1), new solutions are sought that meet both capacitive, efficiency and economic criteria. Increasingly, biodegradable materials, green materials, biomass, waste (including agricultural waste) are used in order to reuse them and reduce greenhouse effects and, above all, to meet the biodegradability criterion necessary for the production of lithium-ion cells as chemical power sources. The most common of these materials are cellulose, starch, wheat, rice, and corn waste, e.g. from agricultural, paper and pharmaceutical production. Such products are subjected to appropriate treatments depending on the desired application (including chemical, thermal, electrochemical). Starch is a biodegradable polysaccharide that consists of polymeric units such as amylose and amylopectin that build an ordered (linear) and amorphous (branched) structure of the polymer. Carbon is also used as a catalyst. Elemental carbon has become available in many nano-structured forms representing the hybridization combinations found in the primary carbon allotropes, and the materials can be enriched with a large number of surface functional groups. There are many examples of catalytic applications of coal in the literature, but the development of this field has been hampered by the lack of a conceptual approach combining structure and function and a lack of understanding of material synthesis. In the context of catalytic applications, the integrity of carbon environmental management properties and parameters such as metal conductivity range and bond sequence management should be characterized. Such data, along with surface and textured information, can form the basis for the provision of network support services.

Keywords: carbon materials, catalysis, BET, capacitors, lithium ion cell

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62 Prevalence and Molecular Characterization of Extended-Spectrum–β Lactamase and Carbapenemase-Producing Enterobacterales from Tunisian Seafood

Authors: Mehdi Soula, Yosra Mani, Estelle Saras, Antoine Drapeau, Raoudha Grami, Mahjoub Aouni, Jean-Yves Madec, Marisa Haenni, Wejdene Mansour

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Multi-resistance to antibiotics in gram-negative bacilli and particularly in enterobacteriaceae, has become frequent in hospitals in Tunisia. However, data on antibiotic resistant bacteria in aquatic products are scarce. The aims of this study are to estimate the proportion of ESBL- and carbapenemase-producing Enterobacterales in seafood (clams and fish) in Tunisia and to molecularly characterize the collected isolates. Two types of seafood were sampled in unrelated markets in four different regions in Tunisia (641 pieces of farmed fish and 1075 mediterranean clams divided into 215 pools, and each pool contained 5 pieces). Once purchased, all samples were incubated in tubes containing peptone salt broth for 24 to 48h at 37°C. After incubation, overnight cultures were isolated on selective MacConkey agar plates supplemented with either imipenem or cefotaxime, identified using API20E test strips (bioMérieux, Marcy-l’Étoile, France) and confirmed by Maldi-TOF MS. Antimicrobial susceptibility was determined by the disk diffusion method on Mueller-Hinton agar plates and results were interpreted according to CA-SFM 2021. ESBL-producing Enterobacterales were detected using the Double Disc Synergy Test (DDST). Carbapenem-resistance was detected using an ertapenem disk and was respectively confirmed using the ROSCO KPC/MBL and OXA-48 Confirm Kit (ROSCO Diagnostica, Taastrup, Denmark). DNA was extracted using a NucleoSpin Microbial DNA extraction kit (Macherey-Nagel, Hoerdt, France), according to the manufacturer’s instructions. Resistance genes were determined using the CGE online tools. The replicon content and plasmid formula were identified from the WGS data using PlasmidFinder 2.0.1 and pMLST 2.0. From farmed fishes, nine ESBL-producing strains (9/641, 1.4%) were isolated, which were identified as E. coli (n=6) and K. pneumoniae (n=3). Among the 215 pools of 5 clams analyzed, 18 ESBL-producing isolates were identified, including 14 E. coli and 4 K. pneumoniae. A low isolation rate of ESBL-producing Enterobacterales was detected 1.6% (18/1075) in clam pools. In fish, the ESBL phenotype was due to the presence of the blaCTX-M-15 gene in all nine isolates, but no carbapenemase gene was identified. In clams, the predominant ESBL phenotype was blaCTX-M-1 (n=6/18). blaCPE (NDM1, OXA48) was detected only in 3 isolates ‘K. pneumoniae isolates’. Replicon typing on the strains carring the ESBL and carbapenemase gene revelead that the major type plasmid carried ESBL were IncF (42.3%) [n=11/26]. In all, our results suggest that seafood can be a reservoir of multi-drug resistant bacteria, most probably of human origin but also by the selection pressure of antibiotic. Our findings raise concerns that seafood bought for consumption may serve as potential reservoirs of AMR genes and pose serious threat to public health.

Keywords: BLSE, carbapenemase, enterobacterales, tunisian seafood

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61 Effects of Temperature and Mechanical Abrasion on Microplastics

Authors: N. Singh, G. K. Darbha

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Since the last decade, a wave of research has begun to study the prevalence and impact of ever-increasing plastic pollution in the environment. The wide application and ubiquitous distribution of plastic have become a global concern due to its persistent nature. The disposal of plastics has emerged as one of the major challenges for waste management landfills. Microplastics (MPs) have found its existence in almost every environment, from the high altitude mountain lake to the deep sea sediments, polar icebergs, coral reefs, estuaries, beaches, and river, etc. Microplastics are fragments of plastics with size less than 5 mm. Microplastics can be classified as primary microplastics and secondary microplastics. Primary microplastics includes purposefully introduced microplastics into the end products for consumers (microbeads used in facial cleansers, personal care product, etc.), pellets (used in manufacturing industries) or fibres (from textile industries) which finally enters into the environment. Secondary microplastics are formed by disintegration of larger fragments under the exposure of sunlight, mechanical abrasive forces by rain, waves, wind and/or water. A number of factors affect the quantity of microplastic present in freshwater environments. In addition to physical forces, human population density proximal to the water body, proximity to urban centres, water residence time, and size of the water body also affects plastic properties. With time, other complex processes in nature such as physical, chemical and biological break down plastics by interfering with its structural integrity. Several studies demonstrate that microplastics found in wastewater sludge being used as manure for agricultural fields, thus having the tendency to alter the soil environment condition influencing the microbial population as well. Inadequate data are available on the fate and transport of microplastics under varying environmental conditions that are required to supplement important information for further research. In addition, microplastics have the tendency to absorb heavy metals and hydrophobic organic contaminants such as PAHs and PCBs from its surroundings and thus acting as carriers for these contaminants in the environment system. In this study, three kinds of microplastics (polyethylene, polypropylene and expanded polystyrene) of different densities were chosen. Plastic samples were placed in sand with different aqueous media (distilled water, surface water, groundwater and marine water). It was incubated at varying temperatures (25, 35 and 40 °C) and agitation levels (rpm). The results show that the number of plastic fragments enhanced with increase in temperature and agitation speed. Moreover, the rate of disintegration of expanded polystyrene is high compared to other plastics. These results demonstrate that temperature, salinity, and mechanical abrasion plays a major role in degradation of plastics. Since weathered microplastics are more harmful as compared to the virgin microplastics, long-term studies involving other environmental factors are needed to have a better understanding of degradation of plastics.

Keywords: environmental contamination, fragmentation, microplastics, temperature, weathering

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60 Development of Alternative Fuels Technologies for Transportation

Authors: Szymon Kuczynski, Krystian Liszka, Mariusz Laciak, Andrii Oliinyk, Adam Szurlej

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Currently, in automotive transport to power vehicles, almost exclusively hydrocarbon based fuels are used. Due to increase of hydrocarbon fuels consumption, quality parameters are tightend for clean environment. At the same time efforts are undertaken for development of alternative fuels. The reasons why looking for alternative fuels for petroleum and diesel are: to increase vehicle efficiency and to reduce the environmental impact, reduction of greenhouse gases emissions and savings in consumption of limited oil resources. Significant progress was performed on development of alternative fuels such as methanol, ethanol, natural gas (CNG / LNG), LPG, dimethyl ether (DME) and biodiesel. In addition, biggest vehicle manufacturers work on fuel cell vehicles and its introduction to the market. Alcohols such as methanol and ethanol create the perfect fuel for spark-ignition engines. Their advantages are high-value antiknock which determines their application as additive (10%) to unleaded petrol and relative purity of produced exhaust gasses. Ethanol is produced in distillation process of plant products, which value as a food can be irrational. Ethanol production can be costly also for the entire economy of the country, because it requires a large complex distillation plants, large amounts of biomass and finally a significant amount of fuel to sustain the process. At the same time, the fermentation process of plants releases into the atmosphere large quantities of carbon dioxide. Natural gas cannot be directly converted into liquid fuels, although such arrangements have been proposed in the literature. Going through stage of intermediates is inevitable yet. Most popular one is conversion to methanol, which can be processed further to dimethyl ether (DME) or olefin (ethylene and propylene) for the petrochemical sector. Methanol uses natural gas as a raw material, however, requires expensive and advanced production processes. In relation to pollution emissions, the optimal vehicle fuel is LPG which is used in many countries as an engine fuel. Production of LPG is inextricably linked with production and processing of oil and gas, and which represents a small percentage. Its potential as an alternative for traditional fuels is therefore proportionately reduced. Excellent engine fuel may be biogas, however, follows to the same limitations as ethanol - the same production process is used and raw materials. Most essential fuel in the campaign of environment protection against pollution is natural gas. Natural gas as fuel may be either compressed (CNG) or liquefied (LNG). Natural gas can also be used for hydrogen production in steam reforming. Hydrogen can be used as a basic starting material for the chemical industry, an important raw material in the refinery processes, as well as a fuel vehicle transportation. Natural gas can be used as CNG which represents an excellent compromise between the availability of the technology that is proven and relatively cheap to use in many areas of the automotive industry. Natural gas can also be seen as an important bridge to other alternative sources of energy derived from fuel and harmless to the environment. For these reasons CNG as a fuel stimulates considerable interest in the worldwide.

Keywords: alternative fuels, CNG (Compressed Natural Gas), LNG (Liquefied Natural Gas), NGVs (Natural Gas Vehicles)

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59 Isolation and Characterization of Chromium Tolerant Staphylococcus aureus from Industrial Wastewater and Their Potential Use to Bioremediate Environmental Chromium

Authors: Muhammad Tariq, Muhammad Waseem, Muhammad Hidayat Rasool

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Isolation and characterization of chromium tolerant Staphylococcus aureus from industrial wastewater and their potential use to bioremediate environmental chromium. Objectives: Chromium with its great economic importance in industrial use is major metal pollutant of the environment. Chromium are used in different industries for various applications such as textile, dyeing and pigmentation, wood preservation, manufacturing pulp and paper, chrome plating, steel and tanning. The release of untreated chromium in industrial effluents causes serious threat to environment and human health, therefore, the current study designed to isolate chromium tolerant Staphylococcus aureus for removal of chromium prior to their final discharge into the environment due to its cost effective and beneficial advantage over physical and chemical methods. Methods: Wastewater samples were collected from discharge point of different industries. Heavy metal analysis by atomic absorption spectrophotometer and microbiological analysis such as total viable count, total coliform, fecal coliform and Escherichia coli were conducted. Staphylococcus aureus was identified through gram’s staining, biomeriux vitek 2 microbial identification system and 16S rRNA gene amplification by polymerase chain reaction. Optimum growth conditions with respect to temperature, pH, salt concentrations and effect of chromium on the growth of bacteria, resistance to other heavy metal ions, minimum inhibitory concentration and chromium uptake ability of Staphylococcus aureus strain K1 was determined by spectrophotometer. Antibiotic sensitivity pattern was also determined by disc diffusion method. Furthermore, chromium uptake ability was confirmed by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope equipped with Oxford Energy Dipersive X-ray (EDX) micro analysis system. Results: The results presented that optimum temperature was 35ᵒC, pH was 8.0 and salt concentration was 0.5% for growth of Staphylococcus aureus K1. The maximum uptake ability of chromium by bacteria was 20mM than other heavy metal ions. The antibiotic sensitivity pattern revealed that Staphylococcus aureus was vancomycin and methicillin sensitive. Non hemolytic activity on blood agar and negative coagulase reaction showed that it was non-pathogenic. Furthermore, the growth of bacteria decreases in the presence of chromium and maximum chromium uptake by bacteria observed at optimum growth conditions. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and Energy dispersive X-ray (EDX) analysis confirmed the presence of chromium uptake by Staphylococcus aureus K1. Conclusion: The study revealed that Staphylococcus aureus K1 have the potential to bio-remediate chromium toxicity from wastewater. Gradually, this biological treatment becomes more important due to its advantage over physical and chemical methods to protect environment and human health.

Keywords: wastewater, staphylococcus, chromium, bioremediation

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58 Investigation of Linezolid, 127I-Linezolid and 131I-Linezolid Effects on Slime Layer of Staphylococcus with Nuclear Methods

Authors: Hasan Demiroğlu, Uğur Avcıbaşı, Serhan Sakarya, Perihan Ünak

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Implanted devices are progressively practiced in innovative medicine to relieve pain or improve a compromised function. Implant-associated infections represent an emerging complication, caused by organisms which adhere to the implant surface and grow embedded in a protective extracellular polymeric matrix, known as a biofilm. In addition, the microorganisms within biofilms enter a stationary growth phase and become phenotypically resistant to most antimicrobials, frequently causing treatment failure. In such cases, surgical removal of the implant is often required, causing high morbidity and substantial healthcare costs. Staphylococcus aureus is the most common pathogen causing implant-associated infections. Successful treatment of these infections includes early surgical intervention and antimicrobial treatment with bactericidal drugs that also act on the surface-adhering microorganisms. Linezolid is a promising anti-microbial with ant-staphylococcal activity, used for the treatment of MRSA infections. Linezolid is a synthetic antimicrobial and member of oxazolidinoni group, with a bacteriostatic or bactericidal dose-dependent antimicrobial mechanism against gram-positive bacteria. Intensive use of antibiotics, have emerged multi-resistant organisms over the years and major problems have begun to be experienced in the treatment of infections occurred with them. While new drugs have been developed worldwide, on the other hand infections formed with microorganisms which gained resistance against these drugs were reported and the scale of the problem increases gradually. Scientific studies about the production of bacterial biofilm increased in recent years. For this purpose, we investigated the activity of Lin, Lin radiolabeled with 131I (131I-Lin) and cold iodinated Lin (127I-Lin) against clinical strains of Staphylococcus aureus DSM 4910 in biofilm. In the first stage, radio and cold labeling studies were performed. Quality-control studies of Lin and iodo (radio and cold) Lin derivatives were carried out by using TLC (Thin Layer Radiochromatography) and HPLC (High Pressure Liquid Chromatography). In this context, it was found that the binding yield was obtained to be about 86±2 % for 131I-Lin. The minimal inhibitory concentration (MIC) of Lin, 127I-Lin and 131I-Lin for Staphylococcus aureus DSM 4910 strain were found to be 1µg/mL. In time-kill studies of Lin, 127I-Lin and 131I-Lin were producing ≥ 3 log10 decreases in viable counts (cfu/ml) within 6 h at 2 and 4 fold of MIC respectively. No viable bacteria were observed within the 24 h of the experiments. Biofilm eradication of S. aureus started with 64 µg/mL of Lin, 127I-Lin and 131I-Lin, and OD630 was 0.507±0.0.092, 0.589±0.058 and 0.266±0.047, respectively. The media control of biofilm producing Staphylococcus was 1.675±0,01 (OD630). 131I and 127I did not have any effects on biofilms. Lin and 127I-Lin were found less effectively than 131I-Lin at killing cells in biofilm and biofilm eradication. Our results demonstrate that the 131I-Lin have potent anti-biofilm activity against S. aureus compare to Lin, 127I-Lin and media control. This is suggested that, 131I may have harmful effect on biofilm structure.

Keywords: iodine-131, linezolid, radiolabeling, slime layer, Staphylococcus

Procedia PDF Downloads 538