Search results for: fish (fluorescence in-situ hybridization)

Authors: Aidan Battison, Neliswa Mama

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Environmental pollution by ionic species has been identified as one of the biggest challenges to the sustainable development of communities. The widespread use of organic and inorganic chemical products and the release of toxic chemical species from industrial waste have resulted in a need for advanced monitoring technologies for environment protection, remediation and restoration. Some of the disadvantages of conventional sensing methods include expensive instrumentation, well-controlled experimental conditions, time-consuming procedures and sometimes complicated sample preparation. On the contrary, the development of fluorescent chemosensors for biological and environmental detection of metal ions has attracted a great deal of attention due to their simplicity, high selectivity, eidetic recognition, rapid response and real-life monitoring. Coumarin derivatives S1 and S2 (Scheme 1) containing 1,2,3-triazole moieties at position -3- have been designed and synthesized from azide and alkyne derivatives by CuAAC “click” reactions for the detection of metal ions. These compounds displayed a strong preference for Fe3+ ions with complexation resulting in fluorescent quenching through photo-induced electron transfer (PET) by the “sphere of action” static quenching model. The tested metal ions included Cd2+, Pb2+, Ag+, Na+, Ca2+, Cr3+, Fe3+, Al3+, Cd2+, Ba2+, Cu2+, Co2+, Hg2+, Zn2+ and Ni2+. The detection limits of S1 and S2 were determined to be 4.1 and 5.1 uM, respectively. Compound S1 displayed the greatest selectivity towards Fe3+ in the presence of competing for metal cations. S1 could also be used for the detection of Fe3+ in a mixture of CH3CN/H¬2¬O. Binding stoichiometry between S1 and Fe3+ was determined by using both Jobs-plot and Benesi-Hildebrand analysis. The binding was shown to occur in a 1:1 ratio between the sensor and a metal cation. Reversibility studies between S1 and Fe3+ were conducted by using EDTA. The binding site of Fe3+ to S1 was determined by using 13 C NMR and Molecular Modelling studies. Complexation was suggested to occur between the lone-pair of electrons from the coumarin-carbonyl and the triazole-carbon double bond.

Keywords: chemosensor, "click" chemistry, coumarin, fluorescence, static quenching, triazole

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Authors: Andrej Pilipović, Branislav Kovačević, Marina Milović, Lazar Kesić, Saša Pekeč, Leopold Poljaković-Pajnik, Saša Orlović

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The effect of landfills on the environment reflects in the dispersion of the contaminants on surrounding soils by the groundwater plume. Such negative effect can be mitigated with the establishment of vegetative buffers surrounding landfills. The “TreeRemEnergy” project funded by the Science Fund of Republic of Serbia – Green program focuses on development of phytobuffers for landfill phytoremediation with the use of Short Rotation Woody Crops (SRWC) plantations that can be further used for the biomass for energy. One of the goals of the project is to select most appropriate poplar (Populus sp.) and willow (Salix sp.) clones through phytorecurrent selection that involves testing of various breeding traits. Physiological parameters serve as a significant contribution to the breeding process aimed to early detection of potential candidates. This study involved testing of the effect of the landfill soils on the photosynthetic processes of the selected poplar and willow candidates. For this purpose, measurements of the gas exchange, chlorophyll content and chlorophyll fluorescence were measured on the tested plants. Obtained results showed that there were differences in the influence of the controlled sources of variation on examined physiological parameters. The effect of clone was significant in all parameters, while the effect of the substrate was not statistically significant in any of measured parameters. However, the effect of interaction Clone×Substrate was significant in intercellular CO2 concentration(ci), stomatal conductance (gs) and transpiration rate (E), suggesting that water regime of the tested clones showed different response to the tested soils. Some clones showed more “generalist” behavior (380, 107/65/9, and PE19/66), while “specialist” behavior was recorded in clones PE4/68, S1-8, and 79/64/2. On the other hand, there was no significant effect of the tested substrate on the pigments content measured with SPAD meter. Results of this study allowed us to narrow the group of clones for further trails in field conditions.

Keywords: clones, net photosynthesis, WUE, transpiration, stomatal conductance, SPAD

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Authors: M. B. Mshelia, J. K. Balogun, J. Auta, N. O. Bankole

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Studies on some aspects of the physico-chemical parameters of Jebba Lake, Niger State, Nigeria was carried out from January to December, 2011. The aim was to investigate some of the physico-chemical parameters relevant to life and health of fish in the water body. Six (6) sampling sites were selected at random which covered Northern (Faku and Awuru), middle (Old Gbajibo and Shankade) and southern zones (New Gbajibo and Jebba dam} of Jebba Lake. Sampling was carried out for the period of 12 Months. The Physico-chemical parameters that were considered were water temperature, pH, dissolved oxygen, electrical conductivity, water transparency, phosphate and nitrate. They were all measured using standard methods. The results showed that water temperature values ranged between 26.06 ± 0.15a in Jebba lake site to 27.34 ± 0.12b in Shankade sampling site, depth varied from 8.08m to 31.64m, water current was between 20.10.62 cm/sec and 26.46 cm/sec, Secchi disc transparency ranged from0.46±0.01 m in New Gbajibo, while the highest mean value was 0.53 ± 0.04 m in Jebba dam., pH varied from 6.49 ± 0.01 and 7.59,5.35±0.03a mg/l in New Gbajibo and 6.75 ± 0.03 mg/l in Faku.The dissolved oxygen varied between 5.35±0.03a mg/l in New Gbajibo and 6.75 ± 0.03 mg/l in Faku.,The mean conductivity value was highest in Faku and Jebba with 128.8 ± 0.32 and 128.8 ± 0.42homs/cm) respectively, Alkalinity ranged 43.00±0.02 to33.30±0.32 mg/l., The nitrate-nitrogen range (2.37 ± 0.08 – 6.40 ± 0.50mg/l)., The mean values of phosphate-phosphorus (PO4-P) recorded varied between 0.18 ± 0.00 mg/l in Faku to 0.47 + 0.10 mg/l in Old Gbajibo.The highest mean value for total dissolved solids was 57.88 ± 0.28 mg/l in Shankade, while the lowest mean value of 39.17 ± 0.42 mg/l was recorded in Faku. Free CO2 ranged from 1.75 mg/l to 2.94 mg/l, Biochemical oxygen demand (BOD) was between 4.25 mg/l and 5.41 mg/l and nitrate-nitrogen concentration was between 2.37 mg/l and 6.40 mg/l. There were significant differences (P < 0.05) between these parameters in relation to stations. Generally, the physico-chemical characteristics of Lake Jebba were within the productive values for aquatic systems, and strongly indicate that the lake is unpolluted.

Keywords: Jebba Lake, water quality, secchi disc, DO meter, sampling sites, physico-chemical parameters

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Authors: Chi Nguyen Thi Quynh, Steven Schilizzi, Atakelty Hailu, Sayed Iftekhar

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Territorial Use Rights for Fisheries (TURFs) have been emerged as a promising tool for fisheries conservation and management. However, illegal fishing has undermined the effectiveness of TURFs, profoundly degrading global fish stocks and marine ecosystems. Conservation and management of fisheries, therefore, largely depends on effectiveness of enforcing fishing regulations, which needs co-enforcement by fishers. However, fishers tend to resist monitoring participation, as their views towards monitoring scheme design has not been received adequate attention. Fishers’ acceptability of a monitoring scheme is likely to be achieved if there is a mechanism allowing fishers to engage in the early planning and design stages. This study carried out a choice experiment with 396 fishers in Vietnam to elicit fishers’ preferences for monitoring scheme and to estimate the relative importance that fishers place on the key design elements. Preference heterogeneity was investigated using a Scale-Adjusted Latent Class Model that accounts for both preference and scale variance. Welfare changes associated with the proposed monitoring schemes were also examined. It is found that there are five distinct preference classes, suggesting that there is no one-size-fits-all scheme well-suited to all fishers. Although fishers prefer to be compensated more for their participation, compensation is not a driving element affecting fishers’ choice. Most fishers place higher value on other elements, such as institutional arrangements and monitoring capacity. Fishers’ preferences are driven by their socio-demographic and psychological characteristics. Understanding of how changes in design elements’ levels affect the participation of fishers could provide policy makers with insights useful for monitoring scheme designs tailored to the needs of different fisher classes.

Keywords: Design of monitoring scheme, Enforcement, Heterogeneity, Illegal Fishing, Territorial Use Rights for Fisheries

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Authors: C. Alexandridou, G. N. Angelopoulos, F. A. Coutelieris

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Construction industry in Greece consumes annually more than 25 million tons of natural aggregates originating mainly from quarries. At the same time, more than 2 million tons of construction and demolition waste are deposited every year, usually without control, therefore increasing the environmental impact of this sector. A potential alternative for saving natural resources and minimize landfilling, could be the recycling and re-use of Concrete and Demolition Waste (CDW) in concrete production. Moreover, in order to conform to the European legislation, Greece is obliged to recycle non-hazardous construction and demolition waste to a minimum of 70% by 2020. In this paper characterization of recycled materials - commercially and laboratory produced, coarse and fine, Recycled Concrete Aggregates (RCA) - has been performed. Namely, X-Ray Fluorescence and X-ray diffraction (XRD) analysis were used for chemical and mineralogical analysis respectively. Physical properties such as particle density, water absorption, sand equivalent and resistance to fragmentation were also determined. This study, first time made in Greece, aims at outlining the differences between RCA and natural aggregates and evaluating their possible influence in concrete performance. Results indicate that RCA’s chemical composition is enriched in Si, Al, and alkali oxides compared to natural aggregates. X-ray diffraction (XRD) analyses results indicated the presence of calcite, quartz and minor peaks of mica and feldspars. From all the evaluated physical properties of coarse RCA, only water absorption and resistance to fragmentation seem to have a direct influence on the properties of concrete. Low Sand Equivalent and significantly high water absorption values indicate that fine fractions of RCA cannot be used for concrete production unless further processed. Chemical properties of RCA in terms of water soluble ions are similar to those of natural aggregates. Four different concrete mixtures were produced and examined, replacing natural coarse aggregates with RCA by a ratio of 0%, 25%, 50% and 75% respectively. Results indicate that concrete mixtures containing recycled concrete aggregates have a minor deterioration of their properties (3-9% lower compression strength at 28 days) compared to conventional concrete containing the same cement quantity.

Keywords: chemical and physical characterization, compressive strength, mineralogical analysis, recycled concrete aggregates, waste management

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Authors: Marita Pigłowska, Beata Kurc, Maciej Galiński

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

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

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Authors: Fuad Abdullah Alatawi

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Microbial infections-based diseases are a significant public health issue around the world, mainly when antibiotic-resistant bacterium types evolve. In this research, we explored the anti-bacterial and anti-cancer potency of iron-oxide (Fe₂O₃) nanoparticles prepared from F. macrocarpa fruit extract. The chemical composition of F. macrocarpa fruit extract was used as a reducing and capping agent for nanoparticles’ synthesis was examined by GC-MS/MS analysis. Then, the prepared nanoparticles were confirmed by various biophysical techniques, including X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), UV-Vis Spectroscopy, and Transmission Electron Microscopy (TEM) and Energy Dispersive Spectroscopy (EDAX), and Dynamic Light Scattering (DLS). Also, the antioxidant capacity of fruit extract was determined through 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Fluorescence Recovery After Photobleaching (FRAP), Superoxide Dismutase (SOD) assays. Furthermore, the cytotoxicity activities of Fe₂O₃ NPs were determined using the (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) test on MCF-7 cells. In the antibacterial assay, lethal doses of the Fe₂O₃NPs effectively inhibited the growth of gram-negative and gram-positive bacteria. The surface damage, ROS production, and protein leakage are the antibacterial mechanisms of Fe₂O₃NPs. Concerning antioxidant activity, the fruit extracts of F. macrocarpa had strong antioxidant properties, which were confirmed by DPPH, ABTS, FRAP, and SOD assays. In addition, the F. microcarpa-derived iron oxide nanomaterials greatly reduced the cell viability of (MCF-7). The GC-MS/MS analysis revealed the presence of 25 main bioactive compounds in the F. microcarpa extract. Overall, the finding of this research revealed that F. microcarpa-derived Fe₂O₃ nanoparticles could be employed as an alternative therapeutic agent to cure microbial infection and breast cancer in humans.

Keywords: ficus microcarpa, iron oxide, antibacterial activity, cytotoxicity

Procedia PDF Downloads 94

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner

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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.

Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag

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Authors: Hafiz Husnain Nawaz

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Anthracnose disease in olive, caused by the fungal pathogen Colletotrichum acutatum, is considered one of the most critical issues in olive orchards in Pakistan. This disease poses a significant threat as it results in infections that can lead to the complete damage of olive plants, affecting leaves, stems, and fruits in the field. Controlling this disease is particularly challenging due to the absence of an effective fungicide that does not pose risks to farmer health and the environment. To address this challenge, our study aimed to evaluate the antagonistic activity of a biosurfactant produced by the Bacillus subtilis PE-07 strain against the anthracnose-causing agent in olive plants. This strain was selected after screening sixty rhizobacteria strains. Additionally, we assessed the heat stability, pH range, and toxicity of the biosurfactant produced by strain PE-07. Our results revealed that the biosurfactant exhibited maximum antifungal activity against C. acutatum. In vitro studies indicated that the biosurfactant could reduce fungal activity by inhibiting the spore germination of C. acutatum. Furthermore, the biosurfactant demonstrated a wide pH and temperature range, displaying antifungal activity at pH levels ranging from 5 to 10 and a temperature range from room temperature to 110°C. To evaluate the biosurfactant's safety, we conducted toxicity tests on zebra fish (Danio rerio). The results showed that the biosurfactant had minimal harmful effects, even at maximum concentrations. In conclusion, our study confirmed that the biosurfactant produced by B. subtilis exhibited high pH and heat stability with minimal harmful effects. Therefore, it presents a promising alternative to chemical pesticides for effectively controlling olive anthracnose in Pakistan.

Keywords: biological control, heat stability and PH range, toxicity, Danio rerio

Procedia PDF Downloads 33

Authors: Dar-Bin Shieh, Gwo-Bin Lee, Chen-Ming Chang, Chen Sheng Yeh, Chih-Chia Huang, Tsung-Ju Li

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Mitochondrial defects have a significant impact in many human diseases and aging associated phenotypes. The pathogenic mitochondrial DNA (mtDNA) mutations are diverse and usually present as heteroplasmic. mtDNA 4977bps deletion is one of the common mtDNA defects, and the ratio of mutated versus normal copy is significantly associated with clinical symptoms thus their quantitative detection has become an important unmet needs for advanced disease diagnosis and therapeutic guidelines. This study revealed a Micro-electro-mechanical-system (MEMS) enabled automatic microfluidic chip that only required minimal sample. The system integrated multiple laboratory operation steps into a Lab-on-a-Chip for high-sensitive and prompt measurement. The entire process including magnetic nanoparticle based mtDNA extraction in chip, mutation selective photonic DNA cleavage, and nanoparticle accelerated photonic quantitative polymerase chain reaction (qPCR). All subsystems were packed inside a miniature three-dimensional micro structured system and operated in an automatic manner. Integration of magnetic beads with microfluidic transportation could promptly extract and enrich the specific mtDNA. The near infrared responsive magnetic nanoparticles enabled micro-PCR to be operated by pulse-width-modulation controlled laser pulsing to amplify the desired mtDNA while quantified by fluorescence intensity captured by a complementary metal oxide system array detector. The proportions of pathogenic mtDNA in total DNA were thus obtained. Micro capillary electrophoresis module was used to analyze the amplicone products. In conclusion, this study demonstrated a new magnetophotonic based qPCR MEMS system that successfully detects and quantify specific disease related DNA mutations thus provides a promising future for rapid diagnosis of mitochondria diseases.

Keywords: mitochondrial DNA, micro-electro-mechanical-system, magnetophotonics, PCR

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Authors: Yushinta Fujaya, Andi Ivo Asphama, Andi Parenrengi, Andi Tenriulo

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Blue Swimming crab (Portunus pelagicus) is an important commercial species throughout the subtropical waters and as such constitutes part of the fisheries resources. Data are lacking on the morphological variations of selected fast-growing crabs reared in a pond. This study aimed to analyze the morphological and molecular character of a selected fast-growing crab reared in ponds in South of Sulawesi. The crab seeds were obtained from local fish-trap and hatchery. A study on the growth was carried out in the population of crabs. The dimensions analyzed were carapace width (CW) measured after 3 months of grow out. Morphological character states were examined based on the pattern of spots on the carapace. Molecular analysis was performed using RAPD (Random Amplified Polymorphic DNA). Genetic distance was analysed using TFPGA (Tools for Population Genetic Analyses) version 1.3. The results showed that there were variations in the growth of crabs. These crabs clustered morphologically into three quite distinct groups. The crab with white spots irregularly spread over its carapace was the largest size while the crab with large white spots scattered over the carapace was the smaller size (3%). The crab with small white spots scattered over the carapace was the smallest size found in this study. Molecular analysis showed that there are morphologically and genetically different between groups of crabs. Genetic distances among crabs ranged from 0.1527 to 0.5856. Thus, this study provides information the use of white spots pattern over carapace as indicators to identify the type of blue swimming crabs.

Keywords: crab, portunus pelagicus, morphology, RAPD, Carapace

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Authors: Sarah El-Gamal, Passent Hossam, Ahmed Abd El Aziz, Rojina Mahmoud, Ahmed Hassan, Dalia Hilal, Eman Ayman, Hana Haytham, Omar Khamis

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Several concerns fall upon the supply chain, especially the cold supply chain. According to the literature, the main challenges in the cold supply chain are the distribution and storage phases. In this research, researchers focused on the storage area, which contains several activities such as the picking activity that faces a lot of obstacles and challenges The implementation of IoT solutions enables businesses to monitor the temperature of food items, which is perhaps the most critical parameter in cold chains. Therefore, researchers proposed a practical solution that would help in eliminating the problems related to ineffective picking for products, especially fish and seafood products, by using IoT technology, most notably LiFi technology. Thus, guaranteeing sufficient picking, reducing waste, and consequently lowering costs. A prototype was specially designed and examined. This research is a single case study research. Two methods of data collection were used; observation and semi-structured interviews. Semi-structured interviews were conducted with managers and decision maker at Carrefour Alexandria to validate the problem and the proposed practical solution using IoTandLiFi technology. A total of three interviews were conducted. As a result, a SWOT analysis was achieved in order to highlight all the strengths and weaknesses of using the recommended Lifi solution in the picking process. According to the investigations, it was found that the use of IoT and LiFi technology is cost effective, efficient, and reduces human errors, minimize the percentage of product waste and thus save money and cost. Thus, increasing customer satisfaction and profits gained.

Keywords: cold supply chain, picking process, temperature control, IOT, warehousing, LIFI

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Authors: Geno Paul S. Cumla, Jan Mariel M. Gentiles, M. Brenda Gajelan-Samson

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Eutrophication is a process where in there is a surplus of nutrients present in a lake or river. Harmful cyanobacteria, hypoxia, and primarily algae, which contain toxins, grow because of the excess nutrients. Algal blooms can cause fish kills, limiting the light penetration which reduces growth of aquatic organisms, causing die-offs of plants and produce conditions that are dangerous to aquatic and human life. The main cause for eutrophication is the presence of excessive amounts of phosphorus (P) and nitrogen (N). Nitrogen is necessary for the production of the plant tissues and is usually used to synthesize proteins. Nitrate is a compound that contains nitrogen, and at elevated levels it can cause harmful effects. Excessive amounts of phosphorus, displaced through human activity, is the major cause of algae growth and as well as degraded water quality. To accomplish this study the Assessment of Soluble inorganic nitrogen (SIN), Assessment of Soluble reactive phosphate (SRP), Determination of Chlorophyll a (Chl-a) concentration, and Determination of Dominating Taxa were done. The study addresses the high probability of algal blooms in Maqueda Bay by assessing the biotic and abiotic factors of Antiao and Jiabong rivers. The data predicts the overgrowth of algae and to create awareness to prevent the event from taking place. The study assesses the adverse effects that could be prevented by understanding and controlling algae. This should predict future cases of algal blooms and allow government agencies which require data to create programs to prevent and assess these issues.

Keywords: eutrophication, chlorophyll a, nitrogen, phosphorus, red tide, Kjeldahl method, spectrophotometer, assessment of soluble inorganic nitrogen, SIN, assessment of soluble reactive phosphate, SRP

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Authors: Mustafa Durmuş, Yesim Ozogul, Esra Balıkcı, Saadet Gokdoğan, Fatih Ozogul, Ali Rıza Köşker, İlknur Yuvka

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Food scientists and researchers have paid attention to develop new ways for improving the nutritional value of foods. The application of nanotechnology techniques to the food industry may allow the modification of food texture, taste, sensory attributes, coloring strength, processability, and stability during shelf life of products. In this research, the effects of nanoemulsions based on commercial oils for vacuum-packed sea bass fillets stored at 2±2°C were investigated in terms of the sensory, chemical (total volatile basic nitrogen (TVB-N), thiobarbituric acid (TBA), peroxide value (PV) and free fatty acids (FFA), pH, water holding capacity (WHC)) and microbiological qualities (total anaerobic bacteria and total lactic acid bacteria). Physical properties of emulsions (viscosity, the particle size of droplet, thermodynamic stability, refractive index, and surface tension) were determined. Nanoemulsion preparation method was based on high energy principle, with ultrasonic homojenizator. Sensory analyses of raw fish showed that the demerit points of the control group were found higher than those of treated groups. The sensory score (odour, taste and texture) of the cooked fillets decreased with storage time, especially in the control. Results obtained from chemical and microbiological analyses also showed that nanoemulsions significantly (p<0.05) decreased the values of biochemical parameters and growth of bacteria during storage period, thus improving quality of vacuum-packed sea bass.

Keywords: quality parameters, nanoemulsion, sea bass, shelf life, vacuum packing

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Authors: Endris Yibru Hanurry, Wei-Hsin Hsu, Hsieh-Chih Tsai

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In recent years, the discovery of small interfering RNAs (siRNAs) has got great attention for the treatment of cancer and other diseases. However, the therapeutic efficacy of siRNAs has been faced with many drawbacks because of short half-life in blood circulation, poor membrane penetration, weak endosomal escape and inadequate release into the cytosol. To overcome these drawbacks, we designed a non-viral vector by conjugating polyamidoamine generation 4.5 dendrimer (PDG4.5) with diethylenetriamine (DETA)- and tetraethylenepentamine (TEPA) followed by binding with siRNA to form polyplexes through electrostatic interaction. The result of 1H nuclear magnetic resonance (NMR), 13C NMR, correlation spectroscopy, heteronuclear single–quantum correlation spectroscopy, and Fourier transform infrared spectroscopy confirmed the successful conjugation of DETA and TEPA with PDG4.5. Then, the size, surface charge, morphology, binding ability, stability, release assay, toxicity and cellular internalization were analyzed to explore the physicochemical and biological properties of PDG4.5-DETA and PDG4.5-TEPA polyplexes at specific N/P ratios. The polyplexes (N/P = 8) exhibited spherical nanosized (125 and 85 nm) particles with optimum surface charge (13 and 26 mV), showed strong siRNA binding ability, protected the siRNA against enzyme digestion and accepted biocompatibility to the HeLa cells. Qualitatively, the fluorescence microscopy image revealed the delocalization (Manders’ coefficient 0.63 and 0.53 for PDG4.5-DETA and PDG4.5-TEPA, respectively) of polyplexes and the translocation of the siRNA throughout the cytosol to show a decent cellular internalization and intracellular biodistribution of polyplexes in HeLa cells. Quantitatively, the flow cytometry result indicated that a significant (P < 0.05) amount of siRNA was internalized by cells treated with PDG4.5-DETA (68.5%) and PDG4.5-TEPA (73%) polyplexes. Generally, PDG4.5-DETA and PDG4.5-TEPA were ideal nanocarriers of siRNA in vitro and might be used as promising candidates for in vivo study and future pharmaceutical applications.

Keywords: non-viral carrier, oligoalkylamine, poly(amidoamine) dendrimer, polyplexes, siRNA

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Authors: Fei Ye, Åsa Barrefelt, Manuchehr Abedi-Valugerdi, Khalid M. Abu-Salah, Salman A. Alrokayan, Mamoun Muhammed, Moustapha Hassan

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With appropriate encapsulation in functional nanoparticles drugs are more stable in physiological environment and the kinetics of the drug can be more carefully controlled and monitored. Furthermore, targeted drug delivery can be developed to improve chemotherapy in cancer treatment, not only by enhancing intracellular uptake by target cells but also by reducing the adverse effects in non-target organs. Inorganic imaging agents, delivered together with anti-cancer drugs, enhance the local imaging contrast and provide precise diagnosis as well as evaluation of therapy efficacy. We have developed biodegradable polymeric vesicles as a nanocarrier system for multimodal bio-imaging and anticancer drug delivery. The poly (lactic-co-glycolic acid) PLGA) vesicles were fabricated by encapsulating inorganic imaging agents of superparamagnetic iron oxide nanoparticles (SPION), manganese-doped zinc sulfide (MN:ZnS) quantum dots (QDs) and the anticancer drug busulfan into PLGA nanoparticles via an emulsion-evaporation method. T2-weighted magnetic resonance imaging (MRI) of PLGA-SPION-Mn:ZnS phantoms exhibited enhanced negative contrast with r2 relaxivity of approximately 523 s-1 mM-1 Fe. Murine macrophage (J774A) cellular uptake of PLGA vesicles started fluorescence imaging at 2 h and reached maximum intensity at 24 h incubation. The drug delivery ability PLGA vesicles was demonstrated in vitro by release of busulfan. PLGA vesicles degradation was studied in vitro, showing that approximately 32% was degraded into lactic and glycolic acid over a period of 5 weeks. The biodistribution of PLGA vesicles was investigated in vivo by MRI in a rat model. Change of contrast in the liver could be visualized by MRI after 7 min and maximal signal loss detected after 4 h post-injection of PLGA vesicles. Histological studies showed that the presence of PLGA vesicles in organs was shifted from the lungs to the liver and spleen over time.

Keywords: biodegradable polymers, multifunctional nanoparticles, quantum dots, anticancer drugs

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Authors: Sospeter P. Maganga, Alphonce Wikedzi, Mussa D. Budeba, Samwel V. Manyele

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This study characterizes complex gold ores (elemental and mineralogical composition, gold distribution, ore grindability, and mineral liberation) for preliminary process selection. About 200 kg of ore samples were collected from each location using systematic sampling by mass interval. Ores were dried, crushed, milled, and split into representative sub-samples (about 1 kg) for elemental and mineralogical composition analyses using X-ray fluorescence (XRF), fire assay finished with Atomic Absorption Spectrometer (AAS), and X-ray Diffraction (XRD) methods, respectively. The gold distribution was studied on size-by-size fractions, while ore grindability was determined using the standard Bond test. The mineral liberation analysis was conducted using ThermoFisher Scientific Mineral Liberation Analyzer (MLA) 650, where unsieved polished grain mounts (80% passing 700 µm) were used as MLA feed. Two MLA measurement modes, X-ray modal analysis (XMOD) and sparse phase liberation-grain X-ray mapping analysis (SPL-GXMAP), were employed. At least two cyanide consumers (Cu, Fe, Pb, and Zn) and kinetics impeders (Mn, S, As, and Bi) were present in all locations investigated. Copper content at Kapanda (0.77% Cu) and Ibindi (7.48% Cu) exceeded the recommended threshold of 0.5% Cu for direct cyanidation. The gold ore at Ibindi indicated a higher rate of grinding compared to other locations. This could be explained by the highest grindability (2.119 g/rev.) and lowest Bond work index (10.213 kWh/t) values. The pyrite-marcasite, chalcopyrite, galena, and siderite were identified as major gold, copper, lead, and iron-bearing minerals, respectively, with potential for economic extraction. However, only gold and copper can be recovered under conventional milling because of grain size issues (galena is exposed by 10%) and process complexity (difficult to concentrate and smelt iron from siderite). Therefore, the preliminary process selection is copper flotation followed by gold cyanidation for Kapanda and Ibindi ores, whereas gold cyanidation with additives such as glycine or ammonia is selected for Mawemeru and Itumbi ores because of low concentrations of Cu, Pb, Fe, and Zn minerals.

Keywords: complex gold ores, mineral liberation, ore characterization, ore grindability

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Authors: Feixiong Chen, Naoufel Haddour, Marie Frenea-Robin, Yves MéRieux, Yann Chevolot, Virginie Monnier

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Neonatal thrombopenia occurs when the mother generates antibodies against her baby’s platelet antigens. It is particularly critical for newborns because it can cause coagulation troubles leading to intracranial hemorrhage. In this case, diagnosis must be done quickly to make platelets transfusion immediately after birth. Before transfusion, platelet antigens must be tested carefully to avoid rejection. The majority of thrombopenia (95 %) are caused by antibodies directed against Human Platelet Antigen 1a (HPA-1a) or 5b (HPA-5b). The common method for antigen platelets detection is polymerase chain reaction allowing for identification of gene sequence. However, it is expensive, time-consuming and requires significant blood volume which is not suitable for newborns. We propose to develop a point-of-care device based on double functionalized magnetic colloids with 1) antibodies specific to antigen platelets and 2) highly sensitive electroactive molecules in order to be detected by an electrochemical microsensor. These magnetic colloids will be used first to isolate platelets from other blood components, then to capture specifically platelets bearing HPA-1a and HPA-5b antigens and finally to attract them close to sensor working electrode for improved electrochemical signal. The expected advantages are an assay time lower than 20 min starting from blood volume smaller than 100 µL. Our functionalization procedure based on amine dendrimers and NHS-ester modification of initial carboxyl colloids will be presented. Functionalization efficiency was evaluated by colorimetric titration of surface chemical groups, zeta potential measurements, infrared spectroscopy, fluorescence scanning and cyclic voltammetry. Our results showed that electroactive molecules and antibodies can be immobilized successfully onto magnetic colloids. Application of a magnetic field onto working electrode increased the detected electrochemical signal. Magnetic colloids were able to capture specific purified antigens extracted from platelets.

Keywords: Magnetic Nanoparticles , Electroactive Molecules, Antibody, Platelet

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Authors: Adam N. Bell, Sarina J. Ergas, Michael Nystrom, Nathan P. Brennan, Kevan L. Main

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Integrated Multi-Trophic Aquaculture systems (IMTA) have the potential to improve the sustainability of seafood production, generate organic fertilizer and feed, remove waste discharges and reduce energy use. IMTA can include periphyton biofilters where algae and microbes grow on surfaces, along with caught detritus and amphipods. Periphyton biofilters provide many advantages: nitrification, denitrification, primary production and ecological diversity. The goal of this study was to determine how biofilter hydraulic residence time (τ) effects periphyton biomass production, dissolved oxygen (DO) and nutrient removal. A pilot scale recirculating aquaculture system (RAS) was designed, constructed and operated at different hydraulic residence times (τ= 1, 2, 4, 6, 8 hours per tank). For each τ, a conservative tracer study was conducted to investigate system hydrodynamics. Data on periphyton weights, pH, nitrogen species, phosphorus, temperature and DO were collected. The tracer study for τ =1 hour revealed that the normalized time < τ, indicating short-circuiting. Periphyton biomass production rate was relatively unaffected by τ (R_e<1 for all τ). Average ammonia nitrogen removal was > 75% for all trials. Nitrate and nitrite did not accumulate in the RAS for τ≥4 hours due to enhanced denitrification in anoxic zones. For τ≥4 hours DO concentration was at a maximum of 4 mg L-1 after 14:00, and decreased to 0 mg L-1 during nighttime. At τ=1 hour, the RAS stayed > 2 mg L-1 and DO was more evenly distributed. For the validation trial, the culture tank was stocked with Centropomus undecimalis (common snook) and the system was operated at τ= 1 hr. Preliminary results showed that a RAS with an integrated periphyton biofilter could support fish health with low nutrient concentrations DO > 6 mg L-1.

Keywords: sustainable aquaculture, resource recovery, nitrogen, microalgae, hydrodynamics, integrated multi-trophic aquaculture

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Authors: Yidan Xu, Pim Martens, Thomas Krafft

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Marine fishing, an energy-intensive activity, directly emits greenhouse gases through fuel combustion, making it a significant contributor to oceanic greenhouse gas (GHG) emissions and worsening climate change. China is the world’s second-largest economy and the top emitter of GHG emissions, and it carries a significant energy conservation and emission reduction burden. However, the increasing GHG emissions from marine fishing is an easily overlooked but essential issue in China. This study offers a diverse perspective by integrating the concepts of total carbon emissions, carbon intensity, and per capita carbon emissions as indicators into calculation and discussion. To better understand the GHG emissions-Gross marine fishery product (GFP) relationship and influencing factors in Chinese marine fishing, the relationship between GHG emissions and economic development in marine fishing, a comprehensive framework is developed by combining the environmental Kuznets curve, the Tapio elasticity index, and the decomposition model. Results indicated that (1) The GHG emissions increased from 16.479 to 18.601 million tons in 2001-2020, in which trawlers and gillnetter are the main source in fishing operation. (2) Total carbon emissions (TC) and CI presented the same decline as GHG emissions, while per capita carbon emissions (PC) displayed an uptrend. (32) GHG emissions and gross marine fishery product (GFP) presented an inverted U-shaped relationship in China; the turning point came in the 13th Five-year Plan period (2016-2020). (43) Most provinces strongly decoupled GFP and CI. Still, PC and TC need more effort to decouple. (54) GHG emissions promoted by an industry structure driven, though carbon intensity and industry scale aid in GHG emissions reduced. (5) Compare with TC and PC, CI has been relatively affected by COVID-19 in 2020. The rise in fish and seafood prices during COVID-19 has boosted the GFP.

Keywords: marine fishing economy, greenhouse gas emission, fishery management, green development

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Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

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Cyanobacteria are gram-negative prokaryotes belonging to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, there two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM), and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods which were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integration sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for the detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly (3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on results, P3HB content in cyanobacteria cells was determined, as also the overall fitness of the cells. Acknowledgment: Funding: This study was partly funded by the projectGA19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), project I 4082-B25.

Keywords: cyanobacteria, fluorescent probe, microscopic techniques, poly(3hydroxybutyrate), spectroscopy, chromatography

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Authors: Hamidullah Binol, Abdullah Bal

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Nowadays, food safety is a great public concern; therefore, robust and effective techniques are required for detecting the safety situation of goods. Hyperspectral Imaging (HSI) is an attractive material for researchers to inspect food quality and safety estimation such as meat quality assessment, automated poultry carcass inspection, quality evaluation of fish, bruise detection of apples, quality analysis and grading of citrus fruits, bruise detection of strawberry, visualization of sugar distribution of melons, measuring ripening of tomatoes, defect detection of pickling cucumber, and classification of wheat kernels. HSI can be used to concurrently collect large amounts of spatial and spectral data on the objects being observed. This technique yields with exceptional detection skills, which otherwise cannot be achieved with either imaging or spectroscopy alone. This paper presents a nonlinear technique based on kernel Fukunaga-Koontz transform (KFKT) for detection of fat content in ground meat using HSI. The KFKT which is the nonlinear version of FKT is one of the most effective techniques for solving problems involving two-pattern nature. The conventional FKT method has been improved with kernel machines for increasing the nonlinear discrimination ability and capturing higher order of statistics of data. The proposed approach in this paper aims to segment the fat content of the ground meat by regarding the fat as target class which is tried to be separated from the remaining classes (as clutter). We have applied the KFKT on visible and nearinfrared (VNIR) hyperspectral images of ground meat to determine fat percentage. The experimental studies indicate that the proposed technique produces high detection performance for fat ratio in ground meat.

Keywords: food (ground meat) inspection, Fukunaga-Koontz transform, hyperspectral imaging, kernel methods

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Authors: E. Feizi Ashtiani, S. Jamshidi, M.H Niksokhan, A. Feizi Ashtiani

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Waste load allocation (WLA) policies may use multi-objective optimization methods to find the most appropriate and sustainable solutions. These usually intend to simultaneously minimize two criteria, total abatement costs (TC) and environmental violations (EV). If other criteria, such as inequity, need for minimization as well, it requires introducing more binary optimizations through different scenarios. In order to reduce the calculation steps, this study presents value index as an innovative decision making approach. Since the value index contains both the environmental violation and treatment costs, it can be maximized simultaneously with the equity index. It implies that the definition of different scenarios for environmental violations is no longer required. Furthermore, the solution is not necessarily the point with minimized total costs or environmental violations. This idea is testified for Haraz River, in north of Iran. Here, the dissolved oxygen (DO) level of river is simulated by Streeter-Phelps equation in MATLAB software. The WLA is determined for fish farms using multi-objective particle swarm optimization (MOPSO) in two scenarios. At first, the trade-off curves of TC-EV and TC-Inequity are plotted separately as the conventional approach. In the second, the Value-Equity curve is derived. The comparative results show that the solutions are in a similar range of inequity with lower total costs. This is due to the freedom of environmental violation attained in value index. As a result, the conventional approach can well be replaced by the value index particularly for problems optimizing these objectives. This reduces the process to achieve the best solutions and may find better classification for scenario definition. It is also concluded that decision makers are better to focus on value index and weighting its contents to find the most sustainable alternatives based on their requirements.

Keywords: waste load allocation (WLA), value index, multi objective particle swarm optimization (MOPSO), Haraz River, equity

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Authors: Zeina Nasr, Bashar Merheb

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The honey bee is a social insect that had driven the human interest much more than any other organism. Beekeeping practices dated the appearance of Man on earth and now it provides a hobby or a secondary work that contributes to the family revenue and requires a little time engagement and money investment. Honey production is not the only contribution of honey bees to the economy, since honey bees play an important role in the pollination. Bee keeping in Lebanon is an important part of the agricultural economy. However, a growing concern about bees is spreading globally, due to an accelerated decline of bees colonies. This raises the alert to preserve and protect local bee strains against uncontrolled introduction of foreign strains and invasive parasitic species. Mitochondrial DNA (mtDNA) markers are commonly used in studying genetic variation in the Apis mellifera species. The DraI-COI-COII test is based on the analysis of the intergenic region between the two genes COI and COII. The different honey bee strains differ in the presence or absence of the p sequence and the number of Q sequences present. A. m. syriaca belonging to the lineage Z, is the native honey bee subspecies in Lebanon, Syria, Jordan, and Palestine. A. m. syriaca is known for its high defensiveness, even though it has many important advantages. However, commercial breeder strains, such as the Italian (A. m. ligustica), and Carniolan (A. m. carnica) strains, have been introduced by beekeepers and regularly used for honey production. This raises worries about the disappearance of the local subspecies. It is obvious that identifying A. m. syriaca colonies and protecting them against uncontrolled mating with other bee strains is a crucial step to protect and improve the original local strain. This study aims to reveal the existing sub-species of honey bee in Akkar – Lebanon and to assess the influence of introgression on the hybridization of the local strain. This will help to identify areas of pure A.m. syriaca population over this district to be considered in choosing syriaca reserves. We collected samples of bees from different regions of Akkar district in order to perform mtDNA analysis. We determined the restriction fragments length of the intergenic region COI-COII, using the restriction enzyme DraI. The results showed both the C and the Z lineages. Four restriction patterns were identified among the restriction maps of the studied samples. The most abundant mitochondrial lineage is the Z lineage constituting about 60% of the identified samples. Al-Dreib region reported the lowest introgression with foreign mtDNA of 21% making it the most suitable area for a genetic reserve of syriaca in Akkar based on its lowest introgression and suitable environment in addition to the attitude of local beekeepers to conserve the local strain. Finally, this study is the first step in constructing conservation programs for the preservation of the local strain and should be generalized to the whole Lebanese population, consistent with the effort done in neighboring countries.

Keywords: Akkar Lebanon, Apis millefera syriaca, DraI-COI-COII test, mitochondrial DNA

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Authors: Renu Sisodia

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This paper attempted to explore and explain the local level river water conflicts in the larger context of state - society relations. This study also covered causes of local level river water conflicts in the catchment area of Bandi and Arvari river of Rajasthan. The focus of the study was on the emergence of community driven, decentralised management of river water bodies and strategies used by local communities to protect and manage river water conflicts. The research is conducted through the process of designing a framework based on essential theoretical and practical findings supported by primary and secondary data. Two in depth case study is conducted to understand the phenomenon in depth. The first field site is Bandi River of Pali district, which is about the struggle between textile industries, community and the State government in which water pollution is said to be one of the driving force of the conflict. Finding shows that the state is supporting textile industries in Pali district have not been adherent to the environmental ethics. Present legal infrastructure and local institutions fail to resolve the serious problem of water pollution in Bandi River and its adverse impact on the local community as a result local community resistance against the local administration and the state government. The second case illustrates the plight of Arvari River in Alwar district. Tussle for the ownership of fisheries between local community, the private fish contractor and State government has been the main bone of contestation. To resolve this conflict local community formed conflict management mechanism named as Arvari Parliament. Arvari Parliament has its own principle and rules to resolve water conflicts related to ownership of the river and use of the river water. The research findings also highlight the co-existence between conventional and modern practices in resolving conflicts.

Keywords: water, water pollution, water conflicts, water scarcity, conflict resolution, local community

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Authors: Patarasuda Chaisupa, R. Clay Wright

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The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.

Keywords: synthetic biology, bioengineering, molecular biology, biotechnology

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Authors: Ashneel Ajay Singh, Naoki Suzuki, Kazumi Sakuramoto, Swastika Roshni, Paras Nath, Alok Kalla

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Albacore tuna (Thunnus alalunga) is one of the commercially important species of tuna in the North Pacific region. Despite the long history of albacore fisheries in the Pacific, its ecological characteristics are not sufficiently understood. The effects of changing climate on numerous commercially and ecologically important fish species including albacore tuna have been documented over the past decades. The objective of this study was to explore and elucidate the relationship of environmental variables with the stock parameters of albacore tuna. The relationship of the North Pacific albacore tuna recruitment (R), spawning stock biomass (SSB) and recruits per spawning biomass (RPS) from 1970 to 2012 with the environmental factors of sea surface temperature (SST), Pacific decadal oscillation (PDO), El Niño southern oscillation (ENSO) and Pacific warm pool index (PWI) was construed. SST and PDO were used as independent variables with SSB to construct stock reproduction models for R and RPS as they showed most significant relationship with the dependent variables. ENSO and PWI were excluded due to collinearity effects with SST and PDO. Model selections were based on R2 values, Akaike Information Criterion (AIC) and significant parameter estimates at p<0.05. Models with single independent variables of SST, PDO, ENSO and PWI were also constructed to illuminate their individual effect on albacore R and RPS. From the results it can be said that SST and PDO resulted in the most significant models for reproducing North Pacific albacore tuna R and RPS time series. SST has the highest impact on albacore R and RPS when comparing models with single environmental variables. It is important for fishery managers and decision makers to incorporate the findings into their albacore tuna management plans for the North Pacific Oceanic region.

Keywords: Albacore tuna, El Niño southern oscillation, Pacific decadal oscillation, sea surface temperature

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Authors: Genesis Julyus T. Agcaoili, Esperanza C. Cabrera

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Thirty (30) isolates of lactic acid bacteria (LAB) from traditionally-prepared fermented products specifically fermented soy-bean paste, fermented mustard and fermented rice-fish mixture were studied for their in vitro antimicrobial and cytotoxic activities. Seventeen (17) isolates were identified as Lactobacillus plantarum, while 13 isolates were identified as Enterococcus spp using 16s rDNA sequences. Disc diffusion method was used to determine the antibacterial activity of LAB against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), while the modified agar overlay method was used to determine the antifungal activity of LAB isolates on the yeast Candida albicans, and the dermatophytes Microsporum gypseum, Trichophyton rubrum and Epidermophyton floccosum. The filter-sterilized LAB supernatants were evaluated for their cytotoxicity to mammalian colon cancer cell lines (HT-29 and HCT116) and normal human dermal fibrolasts (HDFn) using resazurin assay (PrestoBlueTM). Colchicine was the positive control. No antimicrobial activity was observed against the bacterial test organisms and the yeast Candida albicans. On the other hand, all of the tested LAB strains were fungicidal for all the test dermatophytes. Cytotoxicity index profiles of the supernatants of the 15 randomly picked LABs and negative control (brain heart infussion broth) suggest nontoxicity to the cells when compared to colchicine, whereas all LAB supernatants were found to be cytotoxic to HT-29 and HCT116 colon cancer cell lines. Results provide strong support for the role of the lactic acid bacteria studied in antimicrobial treatment and anticancer therapy.

Keywords: antimicrobial, fermented products, fungicidal activity, lactic acid bacteria, probiotics

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Authors: J. Oustric, L. Berti, J. Santini

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Nowadays, the objective of durable agriculture, and in particular organic agriculture, is to reduce the level of fertilizer inputs used in crops. Limiting the quantity of fertilizer inputs would optimize the economical result and minimizing the environmental impact. Nutrient deficiency, particularly of a major nutrient (N, P, and K), can seriously affect fruit production and quality. In citrus crops, rootstock/scion combinations. In citrus crop, scion/rootstock combinations are used frequently to improve tolerance to various abiotic stresses. New rootstocks are needed to respond to these constraints, and the use of new tetraploid rootstocks better adapted to lower nutrient intake could offer a promising way forward. The aim of this work was to determine whether a better tolerance to nutrient deficiency could be observed in a doubled diploid seedling and whether this tolerance could be observed in common clementine scion if used as rootstocks. We selected diploid (CM2x) and doubled diploid (CM4x) Citrumelo 4475 seedlings and common clementine (C) grafted onto Citrumelo 4475 diploid (C/CM2x) and doubled diploid (C/CM4x) rootstocks. Nutrient deficiency effects on the seedlings and scion/rootstock combinations were analyzed by studying anatomical, structural and ultrastructural determinants (chlorosis, stomata, ostiole and cells and their organelles), photosynthetic properties (leaf net photosynthetic rate (Pₙₑₜ), stomatal conductance (gₛ), chlorophyll a fluorescence (Fᵥ/Fₘ)) and oxidative marker (malondialdehyde). Nutrient deficiency affected differently foliar tissues, physiological parameters, and oxidative metabolism in leaves of seedlings depending on their ploidy level and of common clementine scion depending on their rootstocks ploidy level. Both CM4x and C/CM4x presented lower foliar damages (chlorosis, chloroplasts, mitochondria, and plastoglobuli), photosynthesis processes alteration (Pₙₑₜ, gₛ, and Fᵥ/Fₘ), and malondialdehyde accumulation than CM2x and C/CM2x after nutrient deficiency. Doubled diploid Citrumelo 4475 can improve nutrient deficiency tolerance, and its use as a rootstock allows to confer this tolerance to the common clementine scion.

Keywords: nutrient deficiency, oxidative stress, photosynthesis, polyploid rootstocks

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