Search results for: bacterial biomass
Commenced in January 2007
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Edition: International
Paper Count: 2054

Search results for: bacterial biomass

44 Phospholipid Cationic and Zwitterionic Compounds as Potential Non-Toxic Antifouling Agents: A Study of Biofilm Formation Assessed by Micro-titer Assays with Marine Bacteria and Eco-toxicological Effect on Marine Microalgae

Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

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Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro-fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect mammalian eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge or a zwitterionic polar-head group to prevent microfouling with marine bacteria. Toxicity of these compounds was also studied in order to identify the most promising compounds that inhibit biofilm development and show low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, biofilm, marine fouling, non-toxique assays

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43 Efficacy of Sparganium stoloniferum–Derived Compound in the Treatment of Acne Vulgaris: A Pilot Study

Authors: Wanvipa Thongborisute, Punyaphat Sirithanabadeekul, Pichit Suvanprakorn, Anan Jiraviroon

Abstract:

Background: Acne vulgaris is one of the most common dermatologic problems, and can have a significant psychological and physical effect on patients. Propionibacterium acnes' roles in acne vulgaris involve the activation of toll-like receptor 4 (TLR4), and toll-like receptor 2 (TLR2) pathways. By activating these pathways, inflammatory events of acne lesions, comedogenesis and sebaceous lipogenesis can occur. Currently, there are several topical agents commonly use in treating acne vulgaris that are known to have an effect on TLRs, such as retinoic acid and adapalene, but these drugs still have some irritating effects. At present, there is an alarming increase in rate of bacterial resistance due to irrational used of antibiotics both orally and topically. For this reason, acne treatments should contain bioactive molecules targeting at the site of action for the most effective therapeutic effect with the least side effects. Sparganium stoloniferumis a Chinese aquatic herb containing a compound called Sparstolonin B (SsnB), which has been reported to selectively blocks Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4)-mediated inflammatory signals. Therefore, this topical TLR2 and TLR4 antagonist, in a form of Sparganium stoloniferum-derived compound containing SsnB, should give a benefit in reducing inflammation of acne vulgaris lesions and providing an alternative treatments for patients with this condition. Materials and Methods: The objectives of this randomized double blinded split faced placebo controlled trial is to study the safety and efficacy of the Sparganium stoloniferum-derived compound. 32 volunteered patients with mild to moderate degree of acne vulgaris according to global acne grading system were included in the study. After being informed and consented the subjects were given 2 topical treatments for acne vulgaris, one being topical 2.40% Sparganium stoloniferum extraction (containing Sparstolonin B) and the other, placebo. The subjects were asked to apply each treatment to either half of the face daily morning and night by randomization for 8 weeks, and come in for a weekly follow up. For each visit, the patients went through a procedure of lesion counting, including comedones, papules, nodules, pustules, and cystic lesions. Results: During 8 weeks of experimentation, the result shows a reduction in total lesions number between the placebo and the treatment side show statistical significance starting at week 4, where the 95% confidence interval begin to no longer overlap, and shows a trend of continuing to be further apart. The decrease in the amount of total lesions between week 0 and week 8 of the placebo side shows no statistical significant at P value >0.05. While the decrease in the amount of total lesions of acne vulgaris of the treatment side comparing between week 0 and week 8 shows statistical significant at P value <0.001. Conclusion: The data demonstrates that 2.40% Sparganium stoloniferum extraction (containing Sparstolonin B) is more effective in treating acne vulgaris comparing to topical placebo in treating acne vulgaris, by showing significant reduction in the total numbers of acne lesions. Therefore, this topical Sparganium stoloniferum extraction could become a potential alternative treatment for acne vulgaris.

Keywords: acne vulgaris, sparganium stoloniferum, sparstolonin B, toll-like receptor 2, toll-like receptor 4

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42 Comparison of Several Peat Qualities as Amendment to Improve Afforestation of Mine Wastes

Authors: Marie Guittonny-LarchevêQue

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In boreal Canada, industrial activities such as forestry, peat extraction and metal mines often occur nearby. At closure, mine waste storage facilities have to be reclaimed. On tailings storage facilities, tree plantations can achieve rapid restoration of forested landscapes. However, trees poorly grow in mine tailings and organic amendments like peat are required to improve tailings’ structure and nutrients. Canada is a well-known producer of horticultural quality peat, but some lower quality peats coming from areas adjacent to the reclaimed mines could allow successful revegetation. In particular, hemic peat coming from the bottom of peat-bogs is more decomposed than fibric peat and is less valued for horticulture. Moreover, forest peat is sometimes excavated and piled by the forest industry after cuttings to stimulate tree regeneration on the exposed mineral soil. The objective of this project was to compare the ability of peats of differing quality and origin to improve tailings structure, nutrients and tree development. A greenhouse experiment was conducted along one growing season in 2016 with a complete randomized block design combining 8 repetitions (blocks) x 2 tree species (Populus tremuloides and Pinus banksiana) x 6 substrates (tailings, commercial horticultural peat, and mixtures of tailings with commercial peat, forest peat, local fibric peat, or local hemic peat) x 2 fertilization levels (with or without mineral fertilization). The used tailings came from a gold mine and were low in sulfur and trace metals. The commercial peat had a slightly acidic pH (around 6) while other peats had a clearly acidic pH (around 3). However, mixing peat with slightly alkaline tailings resulted in a pH close to 7 whatever the tested peats. The macroporosity of mixtures was intermediate between the low values of tailings (4%) and the high values of commercial peat alone (34%). Seedling survival was lower on tailings for poplar compared to all other treatments, with or without fertilization. Survival and growth were similar among all treatments for pine. Fertilization had no impact on the maximal height and diameter of poplar seedlings but changed the relative performance of the substrates. When not fertilized, poplar seedlings grown in commercial peat were the highest and largest, and the smallest and slenderest in tailings, with intermediate values in mixtures. When fertilized, poplar seedlings grown in commercial peat were smaller and slender compared to all other substrates. However for this species, foliar, shoot, and root biomass production was the greatest in commercial peat and the lowest in tailings compared to all mixtures, whether fertilized or not. The mixture with local fibric peat provided the seedlings with the lowest foliar N concentrations compared to all other substrates whatever the species or the fertilization treatment. At the short-term, the performance of all the tested peats were close when mixed to tailings, showing that peats of lower quality could be valorized instead of using horticultural peat. These results demonstrate that intersectorial synergies in accordance with the principles of circular economy may be developed in boreal Canada between local industries around the reclamation of mine waste dumps.

Keywords: boreal trees, mine spoil, mine revegetation, intersectorial synergies

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41 Investigating the Application of Composting for Phosphorous Recovery from Alum Precipitated and Ferric Precipitated Sludge

Authors: Saba Vahedi, Qiuyan Yuan

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A vast majority of small municipalities and First Nations communities in Manitoba operate facultative or aerated lagoons for wastewater treatment, and most of them use Ferric Chloride (FeCl3) or alum (usually in the form of Al2(SO4)3 ·18H2O) as coagulant for phosphorous removal. The insoluble particles that form during the coagulation process result in a massive volume of sludge which is typically left in the lagoons. Therefore, phosphorous, which is a valuable nutrient, is lost in the process. In this project, the complete recovery of phosphorous from the sludge that is produced in the process of phosphorous removal from wastewater lagoons by using a controlled composting process is investigated. Objective The main objective of this project is to compost alum precipitated sludge that is produced in the process of phosphorous removal in wastewater treatment lagoons in Manitoba. The ultimate goal is to have a product that will meet the characteristics of Class A biosolids in Canada. A number of parameters, including the bioavailability of nutrients in the composted sludge and the toxicity of the sludge, will be evaluated Investigating the bioavailability of phosphorous in the final compost product. The compost will be used as a source of P compared to a commercial fertilizer (monoammonium phosphate MAP) Experimental setup Three different batches of composts piles have been run using the Alum sludge and Ferric sludge. The alum phosphate sludge was collected from an innovative phosphorous removal system at the RM of Taché . The collected sludge was sent to ALS laboratory to analyze the C/N ratio, TP, TN, TC, TAl, moisture contents, pH, and metals concentrations. Wood chips as the bulking agent were collected at the RM of Taché landfill The sludge in the three piles were mixed with 3x dry woodchips. The mixture was turned every week manually. The temperature, the moisture content, and pH were monitored twice a week. The temperature of the mixtures was remained above 55 °C for two weeks. Each pile was kept for ten weeks to get mature. The final products have been applied to two different plants to investigate the bioavailability of P in the compost product as well as the toxicity of the product. The two types of plants were selected based on their sensitivity, growth time, and their compatibility with the Manitoba climate, which are Canola, and switchgrass. The pots are weighed and watered every day to replenish moisture lost by evapotranspiration. A control experiment is also conducted by using topsoil soil and chemical fertilizers (MAP). The experiment will be carried out in a growth room maintained at a day/night temperature regime of 25/15°C, a relative humidity of 60%, and a corresponding photoperiod of 16 h. A total of three cropping (seeding to harvest) cycles need be completed, with each cycle at 50 d in duration. Harvested biomass must be weighed and oven-dried for 72 h at 60°C. The first cycle of growth Canola and Switchgrasses in the alum sludge compost, harvested at the day 50, oven dried, chopped into bits and fine ground in a mill grinder (< 0.2mm), and digested using the wet oxidation method in which plant tissue samples were digested with H2SO4 (99.7%) and H2O2 (30%) in an acid block digester. The digested plant samples need to be analyzed to measure the amount of total phosphorus.

Keywords: wastewater treatment, phosphorus removal, composting alum sludge, bioavailibility of pohosphorus

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40 Kinetic Rate Comparison of Methane Catalytic Combustion of Palladium Catalysts Impregnated onto ɤ-Alumina and Bio-Char

Authors: Noor S. Nasri, Eric C. A. Tatt, Usman D. Hamza, Jibril Mohammed, Husna M. Zain

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Climate change has becoming a global environmental issue that may trigger irreversible changes in the environment with catastrophic consequences for human, animals and plants on our planet. Methane, carbon dioxide and nitrous oxide are the greenhouse gases (GHG) and as the main factor that significantly contributes to the global warming. Mainly carbon dioxide be produced and released to atmosphere by thermal industrial and power generation sectors. Methane is dominant component of natural gas releases significant of thermal heat, and the gaseous pollutants when homogeneous thermal combustion takes place at high temperature. Heterogeneous catalytic Combustion (HCC) principle is promising technologies towards environmental friendly energy production should be developed to ensure higher yields with lower pollutants gaseous emissions and perform complete combustion oxidation at moderate temperature condition as comparing to homogeneous high thermal combustion. Hence the principle has become a very interesting alternative total oxidation for the treatment of pollutants gaseous emission especially NOX product formation. Noble metals are dispersed on a support-porous HCC such as γ- Al2O3, TiO2 and ThO2 to increase thermal stability of catalyst and to increase to effectiveness of catalytic combustion. Support-porous HCC material to be selected based on factors of the surface area, porosity, thermal stability, thermal conductivity, reactivity with reactants or products, chemical stability, catalytic activity, and catalyst life. γ- Al2O3 with high catalytic activity and can last longer life of catalyst, is commonly used as the support for Pd catalyst at low temperatures. Sustainable and renewable support-material of bio-mass char was derived from agro-industrial waste material and used to compare with those the conventional support-porous material. The abundant of biomass wastes generated in palm oil industries is one potential source to convert the wastes into sustainable material as replacement of support material for catalysts. Objective of this study was to compare the kinetic rate of reaction the combustion of methane on Palladium (Pd) based catalyst with Al2O3 support and bio-char (Bc) support derived from shell kernel. The 2wt% Pd was prepared using incipient wetness impregnation method and the HCC performance was accomplished using tubular quartz reactor with gas mixture ratio of 3% methane and 97% air. Material characterization was determined using TGA, SEM, and BET surface area. The methane porous-HCC conversion was carried out by online gas analyzer connected to the reactor that performed porous-HCC. BET surface area for prepared 2 wt% Pd/Bc is smaller than prepared 2wt% Pd/ Al2O3 due to its low porosity between particles. The order of catalyst activity based on kinetic rate on reaction of catalysts in low temperature is prepared 2wt% Pd/Bc > calcined 2wt% Pd/ Al2O3 > prepared 2wt% Pd/ Al2O3 > calcined 2wt% Pd/Bc. Hence the usage of agro-industrial bio-mass waste material can enhance the sustainability principle.

Keywords: catalytic-combustion, environmental, support-bio-char material, sustainable and renewable material

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39 Implementation of Synthesis and Quality Control Procedures of ¹⁸F-Fluoromisonidazole Radiopharmaceutical

Authors: Natalia C. E. S. Nascimento, Mercia L. Oliveira, Fernando R. A. Lima, Leonardo T. C. do Nascimento, Marina B. Silveira, Brigida G. A. Schirmer, Andrea V. Ferreira, Carlos Malamut, Juliana B. da Silva

Abstract:

Tissue hypoxia is a common characteristic of solid tumors leading to decreased sensitivity to radiotherapy and chemotherapy. In the clinical context, tumor hypoxia assessment employing the positron emission tomography (PET) tracer ¹⁸F-fluoromisonidazole ([¹⁸F]FMISO) is helpful for physicians for planning and therapy adjusting. The aim of this work was to implement the synthesis of 18F-FMISO in a TRACERlab® MXFDG module and also to establish the quality control procedure. [¹⁸F]FMISO was synthesized at Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN/Brazil) using an automated synthesizer (TRACERlab® MXFDG, GE) adapted for the production of [¹⁸F]FMISO. The FMISO chemical standard was purchased from ABX. 18O- enriched water was acquired from Center of Molecular Research. Reagent kits containing eluent solution, acetonitrile, ethanol, 2.0 M HCl solution, buffer solution, water for injections and [¹⁸F]FMISO precursor (dissolved in 2 ml acetonitrile) were purchased from ABX. The [¹⁸F]FMISO samples were purified by Solid Phase Extraction method. The quality requirements of [¹⁸F]FMISO are established in the European Pharmacopeia. According to that reference, quality control of [¹⁸F]FMISO should include appearance, pH, radionuclidic identity and purity, radiochemical identity and purity, chemical purity, residual solvents, bacterial endotoxins, and sterility. The duration of the synthesis process was 53 min, with radiochemical yield of (37.00 ± 0.01) % and the specific activity was more than 70 GBq/µmol. The syntheses were reproducible and showed satisfactory results. In relation to the quality control analysis, the samples were clear and colorless at pH 6.0. The spectrum emission, measured by using a High-Purity Germanium Detector (HPGe), presented a single peak at 511 keV and the half-life, determined by the decay method in an activimeter, was (111.0 ± 0.5) min, indicating no presence of radioactive contaminants, besides the desirable radionuclide (¹⁸F). The samples showed concentration of tetrabutylammonium (TBA) < 50μg/mL, assessed by visual comparison to TBA standard applied in the same thin layer chromatographic plate. Radiochemical purity was determined by high performance liquid chromatography (HPLC) and the results were 100%. Regarding the residual solvents tested, ethanol and acetonitrile presented concentration lower than 10% and 0.04%, respectively. Healthy female mice were injected via lateral tail vein with [¹⁸F]FMISO, microPET imaging studies (15 min) were performed after 2 h post injection (p.i), and the biodistribution was analyzed in five-time points (30, 60, 90, 120 and 180 min) after injection. Subsequently, organs/tissues were assayed for radioactivity with a gamma counter. All parameters of quality control test were in agreement to quality criteria confirming that [¹⁸F]FMISO was suitable for use in non-clinical and clinical trials, following the legal requirements for the production of new radiopharmaceuticals in Brazil.

Keywords: automatic radiosynthesis, hypoxic tumors, pharmacopeia, positron emitters, quality requirements

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38 Conservation Challenges of Fish and Fisheries in Lake Tana, Ethiopia

Authors: Shewit Kidane, Abebe Getahun, Wassie Anteneh, Admassu Demeke, Peter Goethals

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We have reviewed major findings of scientific studies on Lake Tana fish resources and their threats. The aim was to provide summarized information for all concerned bodies and international readers to get full and comprehensive picture about the lake’s fish resource and conservation problems. The Lake Tana watershed comprise 28 fish species, of which 21 are endemic. Moreover, Lake Tana is the one among the top 250 lake regions of global importance for biodiversity and it is world recognized migratory birds wintering site. Lake Tana together with its adjacent wetlands provide directly and indirectly a livelihood for more than 500,000 people. However, owing to anthropogenic activities, the lake ecosystem as well as fish and attributes of the fisheries sector are severely degraded. Fish species in Lake Tana are suffering due to illegal fishing, damming, habitat/breeding ground degradation, wastewater disposal, introduction of exotic species, and lack of implementing fisheries regulations. Currently, more than 98% of fishers in Lake Tana are using the most destructive monofilament. Indeed, dams, irrigation schemes and hydropower are constructed in response to the emerging development need only. Mitigation techniques such as construction of fish ladders for the migratory fishes are the most forgotten. In addition, water resource developers are likely unaware of both the importance of the fisheries and the impact of dam construction on fish. As a result, the biodiversity issue is often missed. Besides, Lake Tana wetlands, which play vital role to sustain biodiversity, are not wisely utilised in the sense of the Ramsar Convention’s definition. Wetlands are considered as unhealthy and hence wetland conversion for the purpose of recession agriculture is still seen as advanced mode of development. As a result, many wetlands in the lake watershed are shrinking drastically over time and Cyprus papyrus, one of the characteristic features of Lake Tana, has dramatically declined in its distribution with some local extinction. Furthermore, the recently introduced water hyacinth (Eichhornia crassipes) is creating immense problems on the lake ecosystem. Moreover, currently, 1.56 million tons of sediment have deposited into the lake each year and wastes from the industries and residents are directly discharged into the lake without treatment. Recently, sign of eutrophication is revealed in Lake Tana and most coarsely, the incidence of cyanobacteria genus Microcystis was reported from the Bahir Dar Gulf of Lake Tana. Thus, the direct dependency of the communities on the lake water for drinking as well as to wash their body and clothes and its fisheries make the problem worst. Indeed, since it is home to many endemic migratory fish, such kind of unregulated developmental activities could be detrimental to their stocks. This can be best illustrated by the drastic stock reduction (>75% in biomass) of the world unique Labeobarbus species. So, unless proper management is put in place, the anthropogenic impacts can jeopardize the aquatic ecosystems. Therefore, in order to sustainably use the aquatic resources and fulfil the needs of the local people, every developmental activity and resource utilization should be carried out adhering to the available policies.

Keywords: anthropogenic impacts, dams, endemic fish, wetland degradation

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37 The Importance of Fruit Trees for Prescribed Burning in a South American Savanna

Authors: Rodrigo M. Falleiro, Joaquim P. L. Parime, Luciano C. Santos, Rodrigo D. Silva

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The Cerrado biome is the most biodiverse savanna on the planet. Located in central Brazil, its preservation is seriously threatened by the advance of intensive agriculture and livestock. Conservation Units and Indigenous Lands are increasingly isolated and subject to mega wildfires. Among the characteristics of this savanna, we highlight the high rate of primary biomass production and the reduced occurrence of large grazing animals. In this biome, the predominant fauna is more dependent on the fruits produced by the dicotyledonous species in relation to other tropical savannas. Fire is a key element in the balance between mono and dicotyledons or between the arboreal and herbaceous strata. Therefore, applying fire regimes that maintain the balance between these strata without harming fruit production is essential in the conservation strategies of Cerrado's biodiversity. Recently, Integrated Fire Management has started to be implemented in Brazilian protected areas. As a result, management with prescribed burns has increasingly replaced strategies based on fire exclusion, which in practice have resulted in large wildfires, with highly negative impacts on fruit and fauna production. In the Indigenous Lands, these fires were carried out respecting traditional knowledge. The indigenous people showed great concern about the effects of fire on fruit plants and important animals. They recommended that the burns be carried out between April and May, as it would result in a greater production of edible fruits ("fruiting burning"). In other tropical savannas in the southern hemisphere, the preferential period tends to be later, in the middle of the dry season, when the grasses are dormant (June to August). However, in the Cerrado, this late period coincides with the flowering and sprouting of several important fruit species. To verify the best burning season, the present work evaluated the effects of fire on flowering and fruit production of theByrsonima sp., Mouriri pusa, Caryocar brasiliense, Anacardium occidentale, Pouteria ramiflora, Hancornia speciosa, Byrsonima verbascifolia, Anacardium humille and Talisia subalbens. The evaluations were carried out in the field, covering 31 Indigenous Lands that cover 104,241.18 Km², where 3,386 prescribed burns were carried out between 2015 and 2018. The burning periods were divided into early (carried out during the rainy season), modal or “fruiting” (carried out during the transition between seasons) and late (carried out in the middle of the dry season, when the grasses are dormant). The results corroborate the traditional knowledge, demonstrating that the modal burns result in higher rates of reproduction and fruit production. Late burns showed intermediate results, followed by early burns. We conclude that management strategies based mainly on forage production, which are usually applied in savannas populated by grazing ungulates, may not be the best management strategy for South American savannas. The effects of fire on fruit plants, which have a particular phenologicalsynchronization with the fauna cycle, also need to be observed during the prescription of burns.

Keywords: cerrado biome, fire regimes, native fruits, prescribed burns

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36 A Clinico-Bacteriological Study and Their Risk Factors for Diabetic Foot Ulcer with Multidrug-Resistant Microorganisms in Eastern India

Authors: Pampita Chakraborty, Sukumar Mukherjee

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This study was done to determine the bacteriological profile and antibiotic resistance of the isolates and to find out the potential risk factors for infection with multidrug-resistant organisms. Diabetic foot ulcer is a major medical, social, economic problem and a leading cause of morbidity and mortality, especially in the developing countries like India. 25 percent of all diabetic patients develop a foot ulcer at some point in their lives which is highly susceptible to infections and that spreads rapidly, leading to overwhelming tissue destruction and subsequent amputation. Infection with multidrug resistant organisms (MDRO) may increase the cost of management and may cause additional morbidity and mortality. Proper management of these infections requires appropriate antibiotic selection based on culture and antimicrobial susceptibility testing. Early diagnosis of microbial infections is aimed to institute the appropriate antibacterial therapy initiative to avoid further complications. A total of 200 Type 2 Diabetic Mellitus patients with infection were admitted at GD Hospital and Diabetes Institute, Kolkata. 60 of them who developed ulcer during the year 2013 were included in this study. A detailed clinical history and physical examination were carried out for every subject. Specimens for microbiological studies were obtained from ulcer region. Gram-negative bacilli were tested for extended spectrum Beta-lactamase (ESBL) production by double disc diffusion method. Staphylococcal isolates were tested for susceptibility to oxacillin by screen agar method and disc diffusion. Potential risk factors for MDRO-positive samples were explored. Gram-negative aerobes were most frequently isolated, followed by gram-positive aerobes. Males were predominant in the study and majority of the patients were in the age group of 41-60 years. The presence of neuropathy was observed in 80% cases followed by peripheral vascular disease (73%). Proteus spp. (22) was the most common pathogen isolated, followed by E.coli (17). Staphylococcus aureus was predominant amongst the gram-positive isolates. S.aureus showed a high rate of resistance to antibiotic tested (63.6%). Other gram-positive isolates were found to be highly resistant to erythromycin, tetracycline and ciprofloxacin, 40% each. All isolates were found to be sensitive to Vancomycin and Linezolid. ESBL production was noted in Proteus spp and E.coli. Approximately 70 % of the patients were positive for MDRO. MDRO-infected patients had poor glycemic control (HbA1c 11± 2). Infection with MDROs is common in diabetic foot ulcers and is associated with risk factors like inadequate glycemic control, the presence of neuropathy, osteomyelitis, ulcer size and increased the requirement for surgical treatment. There is a need for continuous surveillance of resistant bacteria to provide the basis for empirical therapy and reduce the risk of complications.

Keywords: diabetic foot ulcer, bacterial infection, multidrug-resistant organism, extended spectrum beta-lactamase

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35 Development of PCL/Chitosan Core-Shell Electrospun Structures

Authors: Hilal T. Sasmazel, Seda Surucu

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Skin tissue engineering is a promising field for the treatment of skin defects using scaffolds. This approach involves the use of living cells and biomaterials to restore, maintain, or regenerate tissues and organs in the body by providing; (i) larger surface area for cell attachment, (ii) proper porosity for cell colonization and cell to cell interaction, and (iii) 3-dimensionality at macroscopic scale. Recent studies on this area mainly focus on fabrication of scaffolds that can closely mimic the natural extracellular matrix (ECM) for creation of tissue specific niche-like environment at the subcellular scale. Scaffolds designed as ECM-like architectures incorporating into the host with minimal scarring/pain and facilitate angiogenesis. This study is related to combining of synthetic PCL and natural chitosan polymers to form 3D PCL/Chitosan core-shell structures for skin tissue engineering applications. Amongst the polymers used in tissue engineering, natural polymer chitosan and synthetic polymer poly(ε-caprolactone) (PCL) are widely preferred in the literature. Chitosan has been among researchers for a very long time because of its superior biocompatibility and structural resemblance to the glycosaminoglycan of bone tissue. However, the low mechanical flexibility and limited biodegradability properties reveals the necessity of using this polymer in a composite structure. On the other hand, PCL is a versatile polymer due to its low melting point (60°C), ease of processability, degradability with non-enzymatic processes (hydrolysis) and good mechanical properties. Nevertheless, there are also several disadvantages of PCL such as its hydrophobic structure, limited bio-interaction and susceptibility to bacterial biodegradation. Therefore, it became crucial to use both of these polymers together as a hybrid material in order to overcome the disadvantages of both polymers and combine advantages of those. The scaffolds here were fabricated by using electrospinning technique and the characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-Ray Photoelectron spectroscopy (XPS). Additionally, gas permeability test, mechanical test, thickness measurement and PBS absorption and shrinkage tests were performed for all type of scaffolds (PCL, chitosan and PCL/chitosan core-shell). By using ImageJ launcher software program (USA) from SEM photographs the average inter-fiber diameter values were calculated as 0.717±0.198 µm for PCL, 0.660±0.070 µm for chitosan and 0.412±0.339 µm for PCL/chitosan core-shell structures. Additionally, the average inter-fiber pore size values exhibited decrease of 66.91% and 61.90% for the PCL and chitosan structures respectively, compare to PCL/chitosan core-shell structures. TEM images proved that homogenous and continuous bead free core-shell fibers were obtained. XPS analysis of the PCL/chitosan core-shell structures exhibited the characteristic peaks of PCL and chitosan polymers. Measured average gas permeability value of produced PCL/chitosan core-shell structure was determined 2315±3.4 g.m-2.day-1. In the future, cell-material interactions of those developed PCL/chitosan core-shell structures will be carried out with L929 ATCC CCL-1 mouse fibroblast cell line. Standard MTT assay and microscopic imaging methods will be used for the investigation of the cell attachment, proliferation and growth capacities of the developed materials.

Keywords: chitosan, coaxial electrospinning, core-shell, PCL, tissue scaffold

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34 Genetically Engineered Crops: Solution for Biotic and Abiotic Stresses in Crop Production

Authors: Deepak Loura

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Production and productivity of several crops in the country continue to be adversely affected by biotic (e.g., Insect-pests and diseases) and abiotic (e.g., water temperature and salinity) stresses. Over-dependence on pesticides and other chemicals is economically non-viable for the resource-poor farmers of our country. Further, pesticides can potentially affect human and environmental safety. While traditional breeding techniques and proper- management strategies continue to play a vital role in crop improvement, we need to judiciously use biotechnology approaches for the development of genetically modified crops addressing critical problems in the improvement of crop plants for sustainable agriculture. Modern biotechnology can help to increase crop production, reduce farming costs, and improve food quality and the safety of the environment. Genetic engineering is a new technology which allows plant breeders to produce plants with new gene combinations by genetic transformation of crop plants for improvement of agronomic traits. Advances in recombinant DNA technology have made it possible to have genes between widely divergent species to develop genetically modified or genetically engineered plants. Plant genetic engineering provides the strength to harness useful genes and alleles from indigenous microorganisms to enrich the gene pool for developing genetically modified (GM) crops that will have inbuilt (inherent) resistance to insect pests, diseases, and abiotic stresses. Plant biotechnology has made significant contributions in the past 20 years in the development of genetically engineered or genetically modified crops with multiple benefits. A variety of traits have been introduced in genetically engineered crops which include (i) herbicide resistance. (ii) pest resistance, (iii) viral resistance, (iv) slow ripening of fruits and vegetables, (v) fungal and bacterial resistance, (vi) abiotic stress tolerance (drought, salinity, temperature, flooding, etc.). (vii) quality improvement (starch, protein, and oil), (viii) value addition (vitamins, micro, and macro elements), (ix) pharmaceutical and therapeutic proteins, and (x) edible vaccines, etc. Multiple genes in transgenic crops can be useful in developing durable disease resistance and a broad insect-control spectrum and could lead to potential cost-saving advantages for farmers. The development of transgenic to produce high-value pharmaceuticals and the edible vaccine is also under progress, which requires much more research and development work before commercially viable products will be available. In addition, molecular-aided selection (MAS) is now routinely used to enhance the speed and precision of plant breeding. Newer technologies need to be developed and deployed for enhancing and sustaining agricultural productivity. There is a need to optimize the use of biotechnology in conjunction with conventional technologies to achieve higher productivity with fewer resources. Therefore, genetic modification/ engineering of crop plants assumes greater importance, which demands the development and adoption of newer technology for the genetic improvement of crops for increasing crop productivity.

Keywords: biotechnology, plant genetic engineering, genetically modified, biotic, abiotic, disease resistance

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33 Functional Traits and Agroecosystem Multifunctionality in Summer Cover Crop Mixtures and Monocultures

Authors: Etienne Herrick

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As an economically and ecologically feasible method for farmers to introduce greater diversity into their crop rotations, cover cropping presents a valuable opportunity for improving the sustainability of food production. Planted in-between cash crop growing seasons, cover crops serve to enhance agroecosystem functioning, rather than being destined for sale or consumption. In fact, cover crops may hold the capacity to deliver multiple ecosystem functions or services simultaneously (multifunctionality). Building upon this line of research will not only benefit society at present, but also support its continued survival through its potential for restoring depleted soils and reducing the need for energy-intensive and harmful external inputs like fertilizers and pesticides. This study utilizes a trait-based approach to explore the influence of inter- and intra-specific interactions in summer cover crop mixtures and monocultures on functional trait expression and ecosystem services. Functional traits that enhance ecosystem services related to agricultural production include height, specific leaf area (SLA), root, shoot ratio, leaf C and N concentrations, and flowering phenology. Ecosystem services include biomass production, weed suppression, reduced N leaching, N recycling, and support of pollinators. Employing a trait-based approach may allow for the elucidation of mechanistic links between plant structure and resulting ecosystem service delivery. While relationships between some functional traits and the delivery of particular ecosystem services may be readily apparent through existing ecological knowledge (e.g. height positively correlating with weed suppression), this study will begin to quantify those relationships so as to gain further understanding of whether and how measurable variation in functional trait expression across cover crop mixtures and monocultures can serve as a reliable predictor of variation in the types and abundances of ecosystem services delivered. Six cover crop species, including legume, grass, and broadleaf functional types, were selected for growth in six mixtures and their component monocultures based upon the principle of trait complementarity. The tricultures (three-way mixtures) are comprised of a legume, grass, and broadleaf species, and include cowpea/sudex/buckwheat, sunnhemp/sudex/buckwheat, and chickling vetch/oat/buckwheat combinations; the dicultures contain the same legume and grass combinations as above, without the buckwheat broadleaf. By combining species with expectedly complimentary traits (for example, legumes are N suppliers and grasses are N acquirers, creating a nutrient cycling loop) the cover crop mixtures may elicit a broader range of ecosystem services than that provided by a monoculture, though trade-offs could exist. Collecting functional trait data will enable the investigation of the types of interactions driving these ecosystem service outcomes. It also allows for generalizability across a broader range of species than just those selected for this study, which may aid in informing further research efforts exploring species and ecosystem functioning, as well as on-farm management decisions.

Keywords: agroecology, cover crops, functional traits, multifunctionality, trait complementarity

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32 The Immunology Evolutionary Relationship between Signal Transducer and Activator of Transcription Genes from Three Different Shrimp Species in Response to White Spot Syndrome Virus Infection

Authors: T. C. C. Soo, S. Bhassu

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Unlike the common presence of both innate and adaptive immunity in vertebrates, crustaceans, in particular, shrimps, have been discovered to possess only innate immunity. This further emphasizes the importance of innate immunity within shrimps in pathogenic resistance. Under the study of pathogenic immune challenge, different shrimp species actually exhibit varying degrees of immune resistance towards the same pathogen. Furthermore, even within the same shrimp species, different batches of challenged shrimps can have different strengths of immune defence. Several important pathways are activated within shrimps during pathogenic infection. One of them is JAK-STAT pathway that is activated during bacterial, viral and fungal infections by which STAT(Signal Transducer and Activator of Transcription) gene is the core element of the pathway. Based on theory of Central Dogma, the genomic information is transmitted in the order of DNA, RNA and protein. This study is focused in uncovering the important evolutionary patterns present within the DNA (non-coding region) and RNA (coding region). The three shrimp species involved are Macrobrachium rosenbergii, Penaeus monodon and Litopenaeus vannamei which all possess commercial significance. The shrimp species were challenged with a famous penaeid shrimp virus called white spot syndrome virus (WSSV) which can cause serious lethality. Tissue samples were collected during time intervals of 0h, 3h, 6h, 12h, 24h, 36h and 48h. The DNA and RNA samples were then extracted using conventional kits from the hepatopancreas tissue samples. PCR technique together with designed STAT gene conserved primers were utilized for identification of the STAT coding sequences using RNA-converted cDNA samples and subsequent characterization using various bioinformatics approaches including Ramachandran plot, ProtParam and SWISS-MODEL. The varying levels of immune STAT gene activation for the three shrimp species during WSSV infection were confirmed using qRT-PCR technique. For one sample, three biological replicates with three technical replicates each were used for qRT-PCR. On the other hand, DNA samples were important for uncovering the structural variations within the genomic region of STAT gene which would greatly assist in understanding the STAT protein functional variations. The partially-overlapping primers technique was used for the genomic region sequencing. The evolutionary inferences and event predictions were then conducted through the Bayesian Inference method using all the acquired coding and non-coding sequences. This was supplemented by the construction of conventional phylogenetic trees using Maximum likelihood method. The results showed that adaptive evolution caused STAT gene sequence mutations between different shrimp species which led to evolutionary divergence event. Subsequently, the divergent sites were correlated to the differing expressions of STAT gene. Ultimately, this study assists in knowing the shrimp species innate immune variability and selection of disease resistant shrimps for breeding purpose. The deeper understanding of STAT gene evolution from the perspective of both purifying and adaptive approaches not only can provide better immunological insight among shrimp species, but also can be used as a good reference for immunological studies in humans or other model organisms.

Keywords: gene evolution, JAK-STAT pathway, immunology, STAT gene

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31 Ecological Planning Method of Reclamation Area Based on Ecological Management of Spartina Alterniflora: A Case Study of Xihu Harbor in Xiangshan County

Authors: Dong Yue, Hua Chen

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The study region Xihu Harbor in Xiangshan County, Ningbo City is located in the central coast of Zhejiang Province. Concerning the wave dispating issue, Ningbo government firstly introduced Spartina alterniflora in 1980s. In the 1990s, S. alterniflora spread so rapidly thus a ‘grassland’ in the sea has been created nowadays. It has become the most important invasive plant of China’s coastal tidal flats. Although S. alterniflora had some ecological and economic functions, it has also brought series of hazards. It has ecological hazards on many aspects, including biomass and biodiversity, hydrodynamic force and sedimentation process, nutrient cycling of tidal flat, succession sequence of soil and plants and so on. On engineering, it courses problems of poor drainage and channel blocking. On economy, the hazard mainly reflected in the threat on aquaculture industry. The purpose of this study is to explore an ecological, feasible and economical way to manage Spartina alterniflora and use the land formed by it, taking Xihu Harbor in Xiangshan County as a case. Comparison method, mathematical modeling, qualitative and quantitative analysis are utilized to proceed the study. Main outcomes are as follows. By comparing a series of S. alterniflora managing methods which include the combination of mechanical cutting and hydraulic reclamation, waterlogging, herbicide and biological substitution from three standpoints – ecology, engineering and economy. It is inferred that the combination of mechanical cutting and hydraulic reclamation is among the top rank of S. alternifora managing methods. The combination of mechanical cutting and hydraulic reclamation means using large-scale mechanical equipment like large screw seagoing dredger to excavate the S. alterniflora with root and mud together. Then the mix of mud and grass was blown off nearby coastal tidal zone transported by pipelines, which can cushion the silt of tidal zone to form a land. However, as man-made land by coast, the reclamation area’s ecological sensitivity is quite high and will face high possibility of flood threat. Therefore, the reclamation area has many reasonability requirements, including ones on location, specific scope, water surface rate, direction of main watercourse, site of water-gate, the ratio of ecological land to urban construction land. These requirements all became important basis when the planning was being made. The water system planning, green space system planning, road structure and land use all need to accommodate the ecological requests. Besides, the profits from the formed land is the managing project’s source of funding, so how to utilize land efficiently is another considered point in the planning. It is concluded that by aiming at managing a large area of S. alterniflora, the combination of mechanical cutting and hydraulic reclamation is an ecological, feasible and economical method. The planning of reclamation area should fully respect the natural environment and possible disasters. Then the planning which makes land use efficient, reasonable, ecological will promote the development of the area’s city construction.

Keywords: ecological management, ecological planning method, reclamation area, Spartina alternifora, Xihu harbor

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30 The Preliminary Exposition of Soil Biological Activity, Microbial Diversity and Morpho-Physiological Indexes of Cucumber under Interactive Effect of Allelopathic Garlic Stalk: A Short-Term Dynamic Response in Replanted Alkaline Soil

Authors: Ahmad Ali, Muhammad Imran Ghani, Haiyan Ding, Zhihui Cheng, Muhammad Iqbal

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Background and Aims: In recent years, protected cultivation trend, especially in the northern parts of China, spread dynamically where production area, structure, and crops diversity have expanded gradually under plastic greenhouse vegetable cropping (PGVC) system. Under this growing system, continuous monoculture with excessive synthetic fertilizers inputs are common cultivation practices frequently adopted by commercial producers. Such long-term cumulative wild exercise year after year sponsor the continuous cropping obstacles in PGVC soil, which have greatly threatened the regional soil eco-sustainability and further impose the continuous assault on soil ecological diversity leading to the exhaustion of agriculture productivity. The aim of this study was to develop new allelopathic insights by exploiting available biological resources in the favor of sustainable PGVC to illuminate the continuous obstacle factors in plastic greenhouse. Method: A greenhouse study was executed under plastic tunnel located at the Horticulture Experimental Station of the College of Horticulture, Northwest A&F University, Yangling, Shaanxi Province, one of the prominent regions for intensive commercial PGVC in China. Post-harvest garlic residues (stalk, leaves) mechanically smashed, homogenized into powder size and incorporated at the ratio of 1:100; 3:100; 5:100 as a soil amendment in a replanted soil that have been used for continuous cucumber monoculture for 7 years (annually double cropping system in a greenhouse). Results: Incorporated C-rich garlic stalk significantly influenced the soil condition through various ways; organic matter decomposition and mineralization, moderately adjusted the soil pH, enhanced the soil nutrient availability, increased enzymatic activities, and promoted 20% more cucumber yield in short-time. Using Illumina MiSeq sequencing analysis of bacterial 16S rRNA and fungal 18S rDNA genes, the current study revealed that addition of garlic stalk/residue could also improve the microbial abundance and community composition in extensively exploited soil, and contributed in soil functionality, caused prosper changes in soil characteristics, reinforced to good crop yield. Conclusion: Our study provided evidence that addition of garlic stalk as soil fertility amendment is a feasible, cost-effective and efficient resource utilization way for renovation of degraded soil health, ameliorate soil quality components and improve ecological environment in short duration. Our study may provide a better scientific understanding for efficient crop residue management typically from allelopathic source.

Keywords: garlic stalk, microbial community dynamics, plant growth, soil amendment, soil-plant system

Procedia PDF Downloads 122
29 Exploitation Pattern of Atlantic Bonito in West African Waters: Case Study of the Bonito Stock in Senegalese Waters

Authors: Ousmane Sarr

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The Senegalese coasts have high productivity of fishery resources due to the frequency of intense up-welling system that occurs along its coast, caused by the maritime trade winds making its waters nutrients rich. Fishing plays a primordial role in Senegal's socioeconomic plans and food security. However, a global diagnosis of the Senegalese maritime fishing sector has highlighted the challenges this sector encounters. Among these concerns, some significant stocks, a priority target for artisanal fishing, need further assessment. If no efforts are made in this direction, most stock will be overexploited or even in decline. It is in this context that this research was initiated. This investigation aimed to apply a multi-modal approach (LBB, Catch-only-based CMSY model and its most recent version (CMSY++); JABBA, and JABBA-Select) to assess the stock of Atlantic bonito, Sarda sarda (Bloch, 1793) in the Senegalese Exclusive Economic Zone (SEEZ). Available catch, effort, and size data from Atlantic bonito over 15 years (2004-2018) were used to calculate the nominal and standardized CPUE, size-frequency distribution, and length at retentions (50 % and 95 % selectivity) of the species. These relevant results were employed as input parameters for stock assessment models mentioned above to define the stock status of this species in this region of the Atlantic Ocean. The LBB model indicated an Atlantic bonito healthy stock status with B/BMSY values ranging from 1.3 to 1.6 and B/B0 values varying from 0.47 to 0.61 of the main scenarios performed (BON_AFG_CL, BON_GN_Length, and BON_PS_Length). The results estimated by LBB are consistent with those obtained by CMSY. The CMSY model results demonstrate that the SEEZ Atlantic bonito stock is in a sound condition in the final year of the main scenarios analyzed (BON, BON-bt, BON-GN-bt, and BON-PS-bt) with sustainable relative stock biomass (B2018/BMSY = 1.13 to 1.3) and fishing pressure levels (F2018/FMSY= 0.52 to 1.43). The B/BMSY and F/FMSY results for the JABBA model ranged between 2.01 to 2.14 and 0.47 to 0.33, respectively. In contrast, The estimated B/BMSY and F/FMSY for JABBA-Select ranged from 1.91 to 1.92 and 0.52 to 0.54. The Kobe plots results of the base case scenarios ranged from 75% to 89% probability in the green area, indicating sustainable fishing pressure and an Atlantic bonito healthy stock size capable of producing high yields close to the MSY. Based on the stock assessment results, this study highlighted scientific advice for temporary management measures. This study suggests an improvement of the selectivity parameters of longlines and purse seines and a temporary prohibition of the use of sleeping nets in the fishery for the Atlantic bonito stock in the SEEZ based on the results of the length-base models. Although these actions are temporary, they can be essential to reduce or avoid intense pressure on the Atlantic bonito stock in the SEEZ. However, it is necessary to establish harvest control rules to provide coherent and solid scientific information that leads to appropriate decision-making for rational and sustainable exploitation of Atlantic bonito in the SEEZ and the Eastern Atlantic Ocean.

Keywords: multi-model approach, stock assessment, atlantic bonito, SEEZ

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28 Explanation of Sentinel-1 Sigma 0 by Sentinel-2 Products in Terms of Crop Water Stress Monitoring

Authors: Katerina Krizova, Inigo Molina

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The ongoing climate change affects various natural processes resulting in significant changes in human life. Since there is still a growing human population on the planet with more or less limited resources, agricultural production became an issue and a satisfactory amount of food has to be reassured. To achieve this, agriculture is being studied in a very wide context. The main aim here is to increase primary production on a spatial unit while consuming as low amounts of resources as possible. In Europe, nowadays, the staple issue comes from significantly changing the spatial and temporal distribution of precipitation. Recent growing seasons have been considerably affected by long drought periods that have led to quantitative as well as qualitative yield losses. To cope with such kind of conditions, new techniques and technologies are being implemented in current practices. However, behind assessing the right management, there is always a set of the necessary information about plot properties that need to be acquired. Remotely sensed data had gained attention in recent decades since they provide spatial information about the studied surface based on its spectral behavior. A number of space platforms have been launched carrying various types of sensors. Spectral indices based on calculations with reflectance in visible and NIR bands are nowadays quite commonly used to describe the crop status. However, there is still the staple limit by this kind of data - cloudiness. Relatively frequent revisit of modern satellites cannot be fully utilized since the information is hidden under the clouds. Therefore, microwave remote sensing, which can penetrate the atmosphere, is on its rise today. The scientific literature describes the potential of radar data to estimate staple soil (roughness, moisture) and vegetation (LAI, biomass, height) properties. Although all of these are highly demanded in terms of agricultural monitoring, the crop moisture content is the utmost important parameter in terms of agricultural drought monitoring. The idea behind this study was to exploit the unique combination of SAR (Sentinel-1) and optical (Sentinel-2) data from one provider (ESA) to describe potential crop water stress during dry cropping season of 2019 at six winter wheat plots in the central Czech Republic. For the period of January to August, Sentinel-1 and Sentinel-2 images were obtained and processed. Sentinel-1 imagery carries information about C-band backscatter in two polarisations (VV, VH). Sentinel-2 was used to derive vegetation properties (LAI, FCV, NDWI, and SAVI) as support for Sentinel-1 results. For each term and plot, summary statistics were performed, including precipitation data and soil moisture content obtained through data loggers. Results were presented as summary layouts of VV and VH polarisations and related plots describing other properties. All plots performed along with the principle of the basic SAR backscatter equation. Considering the needs of practical applications, the vegetation moisture content may be assessed using SAR data to predict the drought impact on the final product quality and yields independently of cloud cover over the studied scene.

Keywords: precision agriculture, remote sensing, Sentinel-1, SAR, water content

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27 Combination of Modelling and Environmental Life Cycle Assessment Approach for Demand Driven Biogas Production

Authors: Juan A. Arzate, Funda C. Ertem, M. Nicolas Cruz-Bournazou, Peter Neubauer, Stefan Junne

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— One of the biggest challenges the world faces today is global warming that is caused by greenhouse gases (GHGs) coming from the combustion of fossil fuels for energy generation. In order to mitigate climate change, the European Union has committed to reducing GHG emissions to 80–95% below the level of the 1990s by the year 2050. Renewable technologies are vital to diminish energy-related GHG emissions. Since water and biomass are limited resources, the largest contributions to renewable energy (RE) systems will have to come from wind and solar power. Nevertheless, high proportions of fluctuating RE will present a number of challenges, especially regarding the need to balance the variable energy demand with the weather dependent fluctuation of energy supply. Therefore, biogas plants in this content would play an important role, since they are easily adaptable. Feedstock availability varies locally or seasonally; however there is a lack of knowledge in how biogas plants should be operated in a stable manner by local feedstock. This problem may be prevented through suitable control strategies. Such strategies require the development of convenient mathematical models, which fairly describe the main processes. Modelling allows us to predict the system behavior of biogas plants when different feedstocks are used with different loading rates. Life cycle assessment (LCA) is a technique for analyzing several sides from evolution of a product till its disposal in an environmental point of view. It is highly recommend to use as a decision making tool. In order to achieve suitable strategies, the combination of a flexible energy generation provided by biogas plants, a secure production process and the maximization of the environmental benefits can be obtained by the combination of process modelling and LCA approaches. For this reason, this study focuses on the biogas plant which flexibly generates required energy from the co-digestion of maize, grass and cattle manure, while emitting the lowest amount of GHG´s. To achieve this goal AMOCO model was combined with LCA. The program was structured in Matlab to simulate any biogas process based on the AMOCO model and combined with the equations necessary to obtain climate change, acidification and eutrophication potentials of the whole production system based on ReCiPe midpoint v.1.06 methodology. Developed simulation was optimized based on real data from operating biogas plants and existing literature research. The results prove that AMOCO model can successfully imitate the system behavior of biogas plants and the necessary time required for the process to adapt in order to generate demanded energy from available feedstock. Combination with LCA approach provided opportunity to keep the resulting emissions from operation at the lowest possible level. This would allow for a prediction of the process, when the feedstock utilization supports the establishment of closed material circles within a smart bio-production grid – under the constraint of minimal drawbacks for the environment and maximal sustainability.

Keywords: AMOCO model, GHG emissions, life cycle assessment, modelling

Procedia PDF Downloads 178
26 Determination of the Presence of Antibiotic Resistance from Vibrio Species in Northern Italy

Authors: Tramuta Clara, Masotti Chiara, Pitti Monica, Adriano Daniela, Battistini Roberta, Serraca Laura, Decastelli Lucia

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Oysters are considered filter organisms, and their raw consumption may increase health risks for consumers: it is often associated with outbreaks of gastroenteritis or enteric illnesses. Most of these foodborne diseases are caused by Vibrio strains, enteric pathogens also involved in the diffusion of genetic determinants of antibiotic resistance and their entrance along the food chain. The European Food Safety Authority (EFSA), during the European Union report on antimicrobial resistance in 2017, focused the attention about the role of food as a possible carrier of antibiotic-resistant bacteria or antibiotic-resistance genes that determine health risks for humans. This study wants to determine antibiotic resistance and antibiotic-resistance genes in Vibrio spp. isolated from Crassostrea gigas oysters collected in the Golfo della Spezia (Liguria, Italy). A total of 47 Vibrio spp. strains were isolated (ISO21872-2:2017) during the summer of 2021 from oysters of Crassostrea gigas. The strains were identified by MALDI-TOF (Bruker, Germany) mass spectrometry and tested for antibiotic susceptibility using a broth microdiluition method (ISO20776-1:2019) using Sensititre EUVSEC plates (Thermo-Fisher Scientific) to obtain the Minimum Inhibitory Concentration (MIC). The strains were tested with PCR-based biomolecular methods, according to previous works, to define the presence of 23 resistance genes of the main classes of antibiotics used in human and veterinary medicine: tet (B), tet (C), tet (D), tet (A), tet (E), tet (G ), tet (K), tet (L), tet (M), tet (O), tet (S) (tetracycline resistance); blaCTX-M, blaTEM, blaOXA, blaSHV (β-lactam resistance); mcr-1 and mcr-2 (colistin resistance); qnrA, qnrB, and qnrS (quinolone resistance); sul1, sul2 and sul3 (sulfonamide resistance). Six different species have been identified: V. alginolyticus 34% (n=16), V. harveyi 28% (n=13), V. fortis 15% (n=7), V. pelagius 8% (n=4), V. parahaemolyticus 11% (n=5) e V. chagasii 4% (n=2). The PCR assays showed the presence of the blaTEM gene on 40% of the strains (n=19). All the other genes were not detected, except for a V. alginolyticus positive for anrS gene. The broth microdiluition method results showed an high level of resistance for ciprofloxacin (62%; n=29), ampicillin (47%; n=22), and colistin (49%; n=23). Furthermore, 32% (n=15) of strains can be considered multiresistant bacteria for the simultaneous presence of resistance for three different antibiotic classes. Susceptibility towards meropenem, azithromycin, gentamicin, ceftazidime, cefotaxime, chloramphenicol, tetracycline and sulphamethoxazole reached 100%. The Vibrio species identified in this study are widespread in marine environments and can cause gastrointerstinal infections after the ingestion of raw fish products and bivalve molluscs. The level of resistance to antibiotics such as ampicillin, ciprofloxacin and colistin can be connected to anthropic factors (industrial, agricultural and domestic wastes) that promote the spread of resistance to these antibiotics. It can be also observed a strong correlation between phenotypic (resistant MIC) and genotypic (positive blaTEM gene) resistance for ampicillin on the same strains, probably due to the transfer of genetic material between bacterial strains. Consumption of raw bivalve molluscs can represent a risk for consumers heath due to the potentially presence of foodborne pathogens, highly resistant to different antibiotics and source of transferable antibiotic-resistant genes.

Keywords: vibrio species, blaTEM genes, antimicrobial resistance, PCR

Procedia PDF Downloads 59
25 In vivo Evaluation of LAB Probiotic Potential with the Zebrafish Animal Model

Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo

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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.

Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish

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24 An Analysis of Economical Drivers and Technical Challenges for Large-Scale Biohydrogen Deployment

Authors: Rouzbeh Jafari, Joe Nava

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This study includes learnings from an engineering practice normally performed on large scale biohydrogen processes. If properly scale-up is done, biohydrogen can be a reliable pathway for biowaste valorization. Most of the studies on biohydrogen process development have used model feedstock to investigate process key performance indicators (KPIs). This study does not intend to compare different technologies with model feedstock. However, it reports economic drivers and technical challenges which help in developing a road map for expanding biohydrogen economy deployment in Canada. BBA is a consulting firm responsible for the design of hydrogen production projects. Through executing these projects, activity has been performed to identify, register and mitigate technical drawbacks of large-scale hydrogen production. Those learnings, in this study, have been applied to the biohydrogen process. Through data collected by a comprehensive literature review, a base case has been considered as a reference, and several case studies have been performed. Critical parameters of the process were identified and through common engineering practice (process design, simulation, cost estimate, and life cycle assessment) impact of these parameters on the commercialization risk matrix and class 5 cost estimations were reported. The process considered in this study is food waste and woody biomass dark fermentation. To propose a reliable road map to develop a sustainable biohydrogen production process impact of critical parameters was studied on the end-to-end process. These parameters were 1) feedstock composition, 2) feedstock pre-treatment, 3) unit operation selection, and 4) multi-product concept. A couple of emerging technologies also were assessed such as photo-fermentation, integrated dark fermentation, and using ultrasound and microwave to break-down feedstock`s complex matrix and increase overall hydrogen yield. To properly report the impact of each parameter KPIs were identified as 1) Hydrogen yield, 2) energy consumption, 3) secondary waste generated, 4) CO2 footprint, 5) Product profile, 6) $/kg-H2 and 5) environmental impact. The feedstock is the main parameter defining the economic viability of biohydrogen production. Through parametric studies, it was found that biohydrogen production favors feedstock with higher carbohydrates. The feedstock composition was varied, by increasing one critical element (such as carbohydrate) and monitoring KPIs evolution. Different cases were studied with diverse feedstock, such as energy crops, wastewater slug, and lignocellulosic waste. The base case process was applied to have reference KPIs values and modifications such as pretreatment and feedstock mix-and-match were implemented to investigate KPIs changes. The complexity of the feedstock is the main bottleneck in the successful commercial deployment of the biohydrogen process as a reliable pathway for waste valorization. Hydrogen yield, reaction kinetics, and performance of key unit operations highly impacted as feedstock composition fluctuates during the lifetime of the process or from one case to another. In this case, concept of multi-product becomes more reliable. In this concept, the process is not designed to produce only one target product such as biohydrogen but will have two or multiple products (biohydrogen and biomethane or biochemicals). This new approach is being investigated by the BBA team and the results will be shared in another scientific contribution.

Keywords: biohydrogen, process scale-up, economic evaluation, commercialization uncertainties, hydrogen economy

Procedia PDF Downloads 91
23 Experimental Study of the Antibacterial Activity and Modeling of Non-isothermal Crystallization Kinetics of Sintered Seashell Reinforced Poly(Lactic Acid) And Poly(Butylene Succinate) Biocomposites Planned for 3D Printing

Authors: Mohammed S. Razali, Kamel Khimeche, Dahah Hichem, Ammar Boudjellal, Djamel E. Kaderi, Nourddine Ramdani

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The use of additive manufacturing technologies has revolutionized various aspects of our daily lives. In particular, 3D printing has greatly advanced biomedical applications. While fused filament fabrication (FFF) technologies have made it easy to produce or prototype various medical devices, it is crucial to minimize the risk of contamination. New materials with antibacterial properties, such as those containing compounded silver nanoparticles, have emerged on the market. In a previous study, we prepared a newly sintered seashell filler (SSh) from bio-based seashells found along the Mediterranean coast using a suitable heat treatment process. We then prepared a series of polylactic acid (PLA) and polybutylene succinate (PBS) biocomposites filled with these SSh particles using a melt mixing technique with a twin-screw extruder to use them as feedstock filaments for 3D printing. The study consisted of two parts: evaluating the antibacterial activity of newly prepared biocomposites made of PLA and PBS reinforced with a sintered seashell in the first part and experimental and modeling analysis of the non-isothermal crystallization kinetics of these biocomposites in the second part. In the first part, the bactericidal activity of the biocomposites against three different bacteria, including Gram-negative bacteria such as (E. coli and Pseudomonas aeruginosa), as well as Gram-positive bacteria such as (Staphylococcus aureus), was examined. The PLA-based biocomposite containing 20 wt.% of SSh particles exhibited an inhibition zone with radial diameters of 8mm and 6mm against E. coli and Pseudo. Au, respectively, while no bacterial activity was observed against Staphylococcus aureus. In the second part, the focus was on investigating the effect of the sintered seashell filler particles on the non-isothermal crystallization kinetics of PLA and PBS 3D-printing composite materials. The objective was to understand the impact of the filler particles on the crystallization mechanism of both PLA and PBS during the cooling process of a melt-extruded filament in (FFF) to manage the dimensional accuracy and mechanical properties of the final printed part. We conducted a non-isothermal melt crystallization kinetic study of a series of PLA-SS and PBS-SS composites using differential scanning calorimetry at various cooling rates. We analyzed the obtained kinetic data using different crystallization kinetic models such as modified Avrami, Ozawa, and Mo's methods. Dynamic mode describes the relative crystallinity as a function of temperature; it found that time half crystallinity (t1/2) of neat PLA decreased from 17 min to 7.3 min for PLA+5 SSh and the (t1/2) of virgin PBS was reduced from 3.5 min to 2.8 min for the composite containing 5wt.% of SSh. We found that the coated SS particles with stearic acid acted as nucleating agents and had a nucleation activity, as observed through polarized optical microscopy. Moreover, we evaluated the effective energy barrier of the non-isothermal crystallization process using the Iso conversional methods of Flynn-Wall-Ozawa (F-W-O) and Kissinger-Akahira-Sunose (K-A-S). The study provides significant insights into the crystallization behavior of PLA and PBS biocomposites.

Keywords: avrami model, bio-based reinforcement, dsc, gram-negative bacteria, gram-positive bacteria, isoconversional methods, non-isothermal crystallization kinetics, poly(butylene succinate), poly(lactic acid), antbactirial activity

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22 Differential Expression Profile Analysis of DNA Repair Genes in Mycobacterium Leprae by qPCR

Authors: Mukul Sharma, Madhusmita Das, Sundeep Chaitanya Vedithi

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Leprosy is a chronic human disease caused by Mycobacterium leprae, that cannot be cultured in vitro. Though treatable with multidrug therapy (MDT), recently, bacteria reported resistance to multiple antibiotics. Targeting DNA replication and repair pathways can serve as the foundation of developing new anti-leprosy drugs. Due to the absence of an axenic culture medium for the propagation of M. leprae, studying cellular processes, especially those belonging to DNA repair pathways, is challenging. Genomic understanding of M. Leprae harbors several protein-coding genes with no previously assigned function known as 'hypothetical proteins'. Here, we report identification and expression of known and hypothetical DNA repair genes from a human skin biopsy and mouse footpads that are involved in base excision repair, direct reversal repair, and SOS response. Initially, a bioinformatics approach was employed based on sequence similarity, identification of known protein domains to screen the hypothetical proteins in the genome of M. leprae, that are potentially related to DNA repair mechanisms. Before testing on clinical samples, pure stocks of bacterial reference DNA of M. leprae (NHDP63 strain) was used to construct standard graphs to validate and identify lower detection limit in the qPCR experiments. Primers were designed to amplify the respective transcripts, and PCR products of the predicted size were obtained. Later, excisional skin biopsies of newly diagnosed untreated, treated, and drug resistance leprosy cases from SIHR & LC hospital, Vellore, India were taken for the extraction of RNA. To determine the presence of the predicted transcripts, cDNA was generated from M. leprae mRNA isolated from clinically confirmed leprosy skin biopsy specimen across all the study groups. Melting curve analysis was performed to determine the integrity of the amplification and to rule out primer‑dimer formation. The Ct values obtained from qPCR were fitted to standard curve to determine transcript copy number. Same procedure was applied for M. leprae extracted after processing a footpad of nude mice of drug sensitive and drug resistant strains. 16S rRNA was used as positive control. Of all the 16 genes involved in BER, DR, and SOS, differential expression pattern of the genes was observed in terms of Ct values when compared to human samples; this was because of the different host and its immune response. However, no drastic variation in gene expression levels was observed in human samples except the nth gene. The higher expression of nth gene could be because of the mutations that may be associated with sequence diversity and drug resistance which suggests an important role in the repair mechanism and remains to be explored. In both human and mouse samples, SOS system – lexA and RecA, and BER genes AlkB and Ogt were expressing efficiently to deal with possible DNA damage. Together, the results of the present study suggest that DNA repair genes are constitutively expressed and may provide a reference for molecular diagnosis, therapeutic target selection, determination of treatment and prognostic judgment in M. leprae pathogenesis.

Keywords: DNA repair, human biopsy, hypothetical proteins, mouse footpads, Mycobacterium leprae, qPCR

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21 Fresh Amnion Membrane Grafting for the Regeneration of Skin in Full Thickness Burn in Newborn - Case Report

Authors: Priyanka Yadav, Umesh Bnasal, Yashvinder Kumar

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The placenta is an important structure that provides oxygen and nutrients to the growing fetus in utero. It is usually thrown away after birth, but it has a therapeutic role in the regeneration of tissue. It is covered by the amniotic membrane, which can be easily separated into the amnion layer and the chorion layer—the amnion layer act as a biofilm for the healing of burn wound and non-healing ulcers. The freshly collected membrane has stem cells, cytokines, growth factors, and anti-inflammatory properties, which act as a biofilm for the healing of wounds. It functions as a barrier and prevents heat and water loss and also protects from bacterial contamination, thus supporting the healing process. The application of Amnion membranes has been successfully used for wound and reconstructive purposes for decades. It is a very cheap and easy process and has shown superior results to allograft and xenograft. However, there are very few case reports of amnion membrane grafting in newborns; we intend to highlight its therapeutic importance in burn injuries in newborns. We present a case of 9 days old male neonate who presented to the neonatal unit of Maulana Azad Medical College with a complaint of fluid-filled blisters and burns wound on the body for six days. He was born outside the hospital at 38 weeks of gestation to a 24-year-old primigravida mother by vaginal delivery. The presentation was cephalic and the amniotic fluid was clear. His birth weight was 2800 gm and APGAR scores were 7 and 8 at 1 and 5 minutes, respectively. His anthropometry was appropriate for gestational age. He developed respiratory distress after birth requiring oxygen support by nasal prongs for three days. On the day of life three, he developed blisters on his body, starting from than face then over the back and perineal region. At a presentation on the day of life nine, he had blisters and necrotic wound on the right side of the face, back, right shoulder and genitalia, affecting 60% of body surface area with full-thickness loss of skin. He was started on intravenous antibiotics and fluid therapy. Pus culture grew Pseudomonas aeuroginosa, for which culture-specific antibiotics were started. Plastic surgery reference was taken and regular wound dressing was done with antiseptics. He had a storming course during the hospital stay. On the day of life 35 when the baby was hemodynamically stable, amnion membrane grafting was done on the wound site; for the grafting, fresh amnion membrane was removed under sterile conditions from the placenta obtained by caesarean section. It was then transported to the plastic surgery unit in half an hour in a sterile fluid where the graft was applied over the infant’s wound. The amnion membrane grafting was done twice in two weeks for covering the whole wound area. After successful uptake of amnion membrane, skin from the thigh region was autografted over the whole wound area by Meek technique in a single setting. The uptake of autograft was excellent and most of the areas were healed. In some areas, there was patchy regeneration of skin so dressing was continued. The infant was discharged after three months of hospital stay and was later followed up in the plastic surgery unit of the hospital.

Keywords: amnion membrane grafting, autograft, meek technique, newborn, regeneration of skin

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20 Chemicals to Remove and Prevent Biofilm

Authors: Cynthia K. Burzell

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Aequor's Founder, a Marine and Medical Microbiologist, discovered novel, non-toxic chemicals in the ocean that uniquely remove biofilm in minutes and prevent its formation for days. These chemicals and over 70 synthesized analogs that Aequor developed can replace thousands of toxic biocides used in consumer and industrial products and, as new drug candidates, kill biofilm-forming bacteria and fungi Superbugs -the antimicrobial-resistant (AMR) pathogens for which there is no cure. Cynthia Burzell, PhD., is a Marine and Medical Microbiologist studying natural mechanisms that inhibit biofilm formation on surfaces in contact with water. In 2002, she discovered a new genus and several new species of marine microbes that produce small molecules that remove biofilm in minutes and prevent its formation for days. The molecules include new antimicrobials that can replace thousands of toxic biocides used in consumer and industrial products and can be developed into new drug candidates to kill the biofilm-forming bacteria and fungi -- including the antimicrobial-resistant (AMR) Superbugs for which there is no cure. Today, Aequor has over 70 chemicals that are divided into categories: (1) Novel natural chemicals. Lonza validated that the primary natural chemical removed biofilm in minutes and stated: "Nothing else known can do this at non-toxic doses." (2) Specialty chemicals. 25 of these structural analogs are already approved under the U.S. Environmental Protection Agency (EPA)'s Toxic Substances Control Act, certified as "green" and available for immediate sale. These have been validated for the following agro-industrial verticals: (a) Surface cleaners: The U.S. Department of Agriculture validated that low concentrations of Aequor's formulations provide deep cleaning of inert, nano and organic surfaces and materials; (b) Water treatments: NASA validated that one dose of Aequor's treatment in the International Space Station's water reuse/recycling system lasted 15 months without replenishment. DOE validated that our treatments lower energy consumption by over 10% in buildings and industrial processes. Future validations include pilot projects with the EPA to test efficacy in hospital plumbing systems. (c) Algae cultivation and yeast fermentation: The U.S. Department of Energy (DOE) validated that Aequor's treatment boosted biomass of renewable feedstocks by 40% in half the time -- increasing the profitability of biofuels and biobased co-products. DOE also validated increased yields and crop protection of algae under cultivation in open ponds. A private oil and gas company validated decontamination of oilfield water. (3) New structural analogs. These kill Gram-negative and Gram-positive bacteria and fungi alone, in combinations with each other, and in combination with low doses of existing, ineffective antibiotics (including Penicillin), "potentiating" them to kill AMR pathogens at doses too low to trigger resistance. Both the U.S. National Institutes for Health (NIH) and Department of Defense (DOD) has executed contracts with Aequor to provide the pre-clinical trials needed for these new drug candidates to enter the regulatory approval pipelines. Aequor seeks partners/licensees to commercialize its specialty chemicals and support to evaluate the optimal methods to scale-up of several new structural analogs via activity-guided fractionation and/or biosynthesis in order to initiate the NIH and DOD pre-clinical trials.

Keywords: biofilm, potentiation, prevention, removal

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19 Isolation and Probiotic Characterization of Lactobacillus plantarum and Lactococcus lactis from Gut Microbiome of Rohu (Labeo rohita)

Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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18 Date Palm Wastes Turning into Biochars for Phosphorus Recovery from Aqueous Solutions: Static and Dynamic Investigations

Authors: Salah Jellali, Nusiba Suliman, Yassine Charabi, Jamal Al-Sabahi, Ahmed Al Raeesi, Malik Al-Wardy, Mejdi Jeguirim

Abstract:

Huge amounts of agricultural biomasses are worldwide produced. At the same time, large quantities of phosphorus are annually discharged into water bodies with possible serious effects onto the environment quality. The main objective of this work is to turn a local Omani biomass (date palm fronds wastes: DPFW) into an effective material for phosphorus recovery from aqueous and the reuse of this P-loaded material in agriculture as ecofriendly amendment. For this aim, the raw DPFW were firstly impregnated with 1 M salt separated solutions of CaCl₂, MgCl₂, FeCl₃, AlCl₃, and a mixture of MgCl₂/AlCl₃ for 24 h, and then pyrolyzed under N2 flow at 500 °C for 2 hours by using an adapted tubular furnace (Carbolite, UK). The synthetized biochars were deeply characterized through specific analyses concerning their morphology, structure, texture, and surface chemistry. These analyses included the use of a scanning electron microscope (SEM) coupled with an energy-dispersive X-Ray spectrometer (EDS), X-Ray diffraction (XRD), Fourier Transform Infrared (FTIR), sorption micrometrics, and X-ray Fluorescence (XRF) apparatus. Then, their efficiency in recovering phosphorus was investigated in batch mode for various contact times (1 min to 3 h), aqueous pH values (from 3 to 11), initial phosphorus concentrations (10-100 mg/L), presence of anions (nitrates, sulfates, and chlorides). In a second step, dynamic assays, by using laboratory columns (height of 30 cm and diameter of 3 cm), were performed in order to investigate the recovery of phosphorus by the modified biochar with a mixture of Mg/Al. The effect of the initial P concentration (25-100 mg/L), the bed depth height (3 to 8 g), and the flow rate (10-30 mL/min) was assessed. Experimental results showed that the biochars physico-chemical properties were very dependent on the type of the used modifying salt. The main affected parameters concerned the specific surface area, microporosity area, and the surface chemistry (pH of zero-point charge and available functional groups). These characteristics have significantly affected the phosphorus recovery efficiency from aqueous solutions. Indeed, the P removal efficiency in batch mode varies from about 5 mg/g for the Fe-modified biochar to more than 13 mg/g for the biochar functionalized with Mg/Al layered double hydroxides. Moreover, the P recovery seems to be a time dependent process and significantly affected by the pH of the aqueous media and the presence of foreign anions due to competition phenomenon. The laboratory column study of phosphorus recovery by the biochar functionalized with Mg/Al layered double hydroxides showed that this process is affected by the used phosphorus concentration, the flow rate, and especially the column bed depth height. Indeed, the phosphorus recovered amount increased from about 4.9 to more than 9.3 mg/g used biochar mass of 3 and 8 g, respectively. This work proved that salt-modified palm fronds-derived biochars could be considered as attractive and promising materials for phosphorus recovery from aqueous solutions even under dynamic conditions. The valorization of these P-loaded-modified biochars as eco-friendly amendment for agricultural soils is necessary will promote sustainability and circular economy concepts in the management of both liquid and solid wastes.

Keywords: date palm wastes, Mg/Al double-layered hydroxides functionalized biochars, phosphorus, recovery, sustainability, circular economy

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17 High Purity Lignin for Asphalt Applications: Using the Dawn Technology™ Wood Fractionation Process

Authors: Ed de Jong

Abstract:

Avantium is a leading technology development company and a frontrunner in renewable chemistry. Avantium develops disruptive technologies that enable the production of sustainable high value products from renewable materials and actively seek out collaborations and partnerships with like-minded companies and academic institutions globally, to speed up introductions of chemical innovations in the marketplace. In addition, Avantium helps companies to accelerate their catalysis R&D to improve efficiencies and deliver increased sustainability, growth, and profits, by providing proprietary systems and services to this regard. Many chemical building blocks and materials can be produced from biomass, nowadays mainly from 1st generation based carbohydrates, but potential for competition with the human food chain leads brand-owners to look for strategies to transition from 1st to 2nd generation feedstock. The use of non-edible lignocellulosic feedstock is an equally attractive source to produce chemical intermediates and an important part of the solution addressing these global issues (Paris targets). Avantium’s Dawn Technology™ separates the glucose, mixed sugars, and lignin available in non-food agricultural and forestry residues such as wood chips, wheat straw, bagasse, empty fruit bunches or corn stover. The resulting very pure lignin is dense in energy and can be used for energy generation. However, such a material might preferably be deployed in higher added value applications. Bitumen, which is fossil based, are mostly used for paving applications. Traditional hot mix asphalt emits large quantities of the GHG’s CO₂, CH₄, and N₂O, which is unfavorable for obvious environmental reasons. Another challenge for the bitumen industry is that the petrochemical industry is becoming more and more efficient in breaking down higher chain hydrocarbons to lower chain hydrocarbons with higher added value than bitumen. This has a negative effect on the availability of bitumen. The asphalt market, as well as governments, are looking for alternatives with higher sustainability in terms of GHG emission. The usage of alternative sustainable binders, which can (partly) replace the bitumen, contributes to reduce GHG emissions and at the same time broadens the availability of binders. As lignin is a major component (around 25-30%) of lignocellulosic material, which includes terrestrial plants (e.g., trees, bushes, and grass) and agricultural residues (e.g., empty fruit bunches, corn stover, sugarcane bagasse, straw, etc.), it is globally highly available. The chemical structure shows resemblance with the structure of bitumen and could, therefore, be used as an alternative for bitumen in applications like roofing or asphalt. Applications such as the use of lignin in asphalt need both fundamental research as well as practical proof under relevant use conditions. From a fundamental point of view, rheological aspects, as well as mixing, are key criteria. From a practical point of view, behavior in real road conditions is key (how easy can the asphalt be prepared, how easy can it be applied on the road, what is the durability, etc.). The paper will discuss the fundamentals of the use of lignin as bitumen replacement as well as the status of the different demonstration projects in Europe using lignin as a partial bitumen replacement in asphalts and will especially present the results of using Dawn Technology™ lignin as partial replacement of bitumen.

Keywords: biorefinery, wood fractionation, lignin, asphalt, bitumen, sustainability

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16 A Multi-Model Approach to Assess Atlantic Bonito (Sarda Sarda, Bloch 1793) in the Eastern Atlantic Ocean: A Case Study of the Senegalese Exclusive Economic Zone

Authors: Ousmane Sarr

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The Senegalese coasts have high productivity of fishery resources due to the frequency of intense up-welling system that occurs along its coast, caused by the maritime trade winds making its waters nutrients rich. Fishing plays a primordial role in Senegal's socioeconomic plans and food security. However, a global diagnosis of the Senegalese maritime fishing sector has highlighted the challenges this sector encounters. Among these concerns, some significant stocks, a priority target for artisanal fishing, need further assessment. If no efforts are made in this direction, most stock will be overexploited or even in decline. It is in this context that this research was initiated. This investigation aimed to apply a multi-modal approach (LBB, Catch-only-based CMSY model and its most recent version (CMSY++); JABBA, and JABBA-Select) to assess the stock of Atlantic bonito, Sarda sarda (Bloch, 1793) in the Senegalese Exclusive Economic Zone (SEEZ). Available catch, effort, and size data from Atlantic bonito over 15 years (2004-2018) were used to calculate the nominal and standardized CPUE, size-frequency distribution, and length at retentions (50 % and 95 % selectivity) of the species. These relevant results were employed as input parameters for stock assessment models mentioned above to define the stock status of this species in this region of the Atlantic Ocean. The LBB model indicated an Atlantic bonito healthy stock status with B/BMSY values ranging from 1.3 to 1.6 and B/B0 values varying from 0.47 to 0.61 of the main scenarios performed (BON_AFG_CL, BON_GN_Length, and BON_PS_Length). The results estimated by LBB are consistent with those obtained by CMSY. The CMSY model results demonstrate that the SEEZ Atlantic bonito stock is in a sound condition in the final year of the main scenarios analyzed (BON, BON-bt, BON-GN-bt, and BON-PS-bt) with sustainable relative stock biomass (B2018/BMSY = 1.13 to 1.3) and fishing pressure levels (F2018/FMSY= 0.52 to 1.43). The B/BMSY and F/FMSY results for the JABBA model ranged between 2.01 to 2.14 and 0.47 to 0.33, respectively. In contrast, The estimated B/BMSY and F/FMSY for JABBA-Select ranged from 1.91 to 1.92 and 0.52 to 0.54. The Kobe plots results of the base case scenarios ranged from 75% to 89% probability in the green area, indicating sustainable fishing pressure and an Atlantic bonito healthy stock size capable of producing high yields close to the MSY. Based on the stock assessment results, this study highlighted scientific advice for temporary management measures. This study suggests an improvement of the selectivity parameters of longlines and purse seines and a temporary prohibition of the use of sleeping nets in the fishery for the Atlantic bonito stock in the SEEZ based on the results of the length-base models. Although these actions are temporary, they can be essential to reduce or avoid intense pressure on the Atlantic bonito stock in the SEEZ. However, it is necessary to establish harvest control rules to provide coherent and solid scientific information that leads to appropriate decision-making for rational and sustainable exploitation of Atlantic bonito in the SEEZ and the Eastern Atlantic Ocean.

Keywords: multi-model approach, stock assessment, atlantic bonito, healthy stock, sustainable, SEEZ, temporary management measures

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15 Gene Cloning and Expression of Azoreductases from Azo-Degraders Lysinibacillus macrolides and Bacillus coagulans Isolated from Egyptian Industrial Wastewater

Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

Procedia PDF Downloads 116