Search results for: maritimus species

Authors: Cho Achidi

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Tomato is a crucial food crop significantly contributes to global food and nutrition security. However, postharvest losses severely limit its role. Therefore, it is necessary to develop sustainable strategies to minimize these losses and improve the shelf-life of tomato fruits. One of the major concerns is bacterial infections, particularly by faecal coliform bacteria, which can cause food poisoning and illnesses like diarrhoea and dysentery. This study seeks to identify the presence of coliform bacteria on tomato fruits in fields and markets in Muea, Buea Municipality. The study also evaluated different management strategies to reduce the bacterial incidence and load on tomato fruits. A total of 200 fruits were sampled for both the coliform survey and shelf-life analysis. Ten farmers and traders provided samples, including asymptomatic and symptomatic tomato fruits. The samples designated for shelf-life analysis were treated with Aquatab, warm water, lemon, and onion. The results indicated that out of the 80 symptomatic samples collected, 12.5% contained faecal and total coliform species. Among the ten farms sampled, 14% were infected with coliform bacteria, with the highest infestation rate of 60% recorded in field 4. Furthermore, 15% of the asymptomatic tomato fruits were found to be infected by coliform bacteria. Regarding the management strategies, Aquatabs exhibited the highest efficacy in reducing the incidence of coliform bacteria on tomato fruits, followed by onion and lemon extracts. Although hot water treatment effectively removed bacteria from the fruits, damaging the cell wall negatively affected their shelf-life. Overall, this study emphasizes the severity of coliform bacterial pathogens in the Muea area, particularly their occurrence on asymptomatic tomatoes, which poses a significant concern for plant quarantine services. It also demonstrates potential options for mitigating this bacterial challenge.

Keywords: tomato, shelf-life analysis, food and nutrition security, coliform bbacteria

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Authors: Suresh Kumar Verma, Jugal Kishore Das, Ealisha Jha, Mrutyunjay Suar, SKS Parashar

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In recent years nanoforms of inorganic metallic oxides has attracted a lot of interest due to their small size and significantly improved physical, chemical and biological properties compared to their molecular precursor. Some of the inorganic materials such as TiO2, ZnO, MgO, CaO, Al2O3 have been extensively used in biological applications. Zinc Oxide is a Wurtzite-type semiconductor and piezo-electric material exhibiting excellent electrical, optical and chemical properties with a band energy gap of 3.1-3.4 eV. Nanoforms of Zinc Oxide (ZnO) are increasingly recognised for their utility in biological application. The significant physical parameters such as surface area, particle size, surface charge and Zeta potential of Zinc Oxide (ZnO) nanoparticles makes it suitable for the uptake, persistance, biological, and chemical activities inside the living cells. The present study shows the effect of intrinsic defects of ZnO nanocrystals synthesized by high energy ball milling (HEBM) technique in their antibacterial activities. Bulk Zinc oxide purchased from market were ball milled for 7 h, 10 h, and 15 h respectively to produce nanosized Zinc Oxide. The structural and optical modification of such synthesized particles were determined by X-ray diffraction (XRD), Scanning Electron Microscopy and Electron Paramagnetic Resonance (EPR). The antibacterial property of synthesized Zinc Oxide nanoparticles was tested using well diffusion, minimum inhibitory Concentration, minimum bacteriocidal concentration, reactive oxygen species (ROS) estimation and membrane potential determination methods. In this study we observed that antibacterial activity of ZnO nanoparticles is because of the intrinsic defects that exist as a function of difference in size and milling time.

Keywords: high energy ball milling, ZnO nanoparticles, EPR, Antibacterial properties

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Authors: Wala Abou Saoud, Aymen Amine Assadi, Monia Guiza, Abdelkrim Bouzaza, Wael Aboussaoud, Abdelmottaleb Ouederni, Dominique Wolbert

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Volatile organic compounds (VOCs) constitute one of the most important families of chemicals involved in atmospheric pollution, causing damage to the environment and human health, and need, consequently, to be eliminated. Among the promising technologies, dielectric barrier discharge (DBD) plasma - photocatalysis coupling reveals very interesting prospects in terms of process synergy of compounds mineralization’s, with low energy consumption. In this study, the removal of organic compounds such butyraldehyde (BUTY) and dimethyl disulfide (DMDS) (exhaust gasses from animal quartering centers.) in air mixture using DBD plasma coupled with photocatalysis was tested, in order to determine whether or not synergy effect was present. The removal efficiency of these pollutants, a selectivity of CO₂ and CO, and byproducts formation such as ozone formation were investigated in order to evaluate the performance of the combined process. For this purpose, a series of experiments were carried out in a continuous reactor. Many operating parameters were also investigated such as the specific energy of discharge, the inlet concentration of pollutant and the flowrate. It appears from this study that, the performance of the process has enhanced and a synergetic effect is observed. In fact, we note an enhancement of 10 % on removal efficiency. It is interesting to note that the combined system leads to better CO₂ selectivity than for plasma. Consequently, intermediates by-products have been reduced due to various other species (O•, N, OH•, O₂•-, O₃, NO₂, NOx, etc.). Additionally, the behavior of combining DBD plasma and photocatalysis has shown that the ozone can be easily also decomposed in presence of photocatalyst.

Keywords: combined process, DBD plasma, photocatalysis, pilot scale, synergetic effect, VOCs

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Authors: Yongjiang Zhang, Kallol Barai, Umesh R. Hodeghatta, Trang Tran, Vikas Dhiman

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Detecting water stress on crops early and accurately is crucial to minimize its impact. This study aims to measure water stress in wild blueberry crops non-destructively by analyzing proximal hyperspectral data. The data collection took place in the summer growing season of 2022. A drought experiment was conducted on wild blueberries in the randomized block design in the greenhouse, incorporating various genotypes and irrigation treatments. Hyperspectral data ( spectral range: 400-1000 nm) using a handheld spectroradiometer and leaf water potential data using a pressure chamber were collected from wild blueberry plants. Machine learning techniques, including multiple regression analysis and random forest models, were employed to predict leaf water potential (MPa). We explored the optimal wavelength bands for simple differences (RY1-R Y2), simple ratios (RY1/RY2), and normalized differences (|RY1-R Y2|/ (RY1-R Y2)). NDWI ((R857 - R1241)/(R857 + R1241)), SD (R2188 – R2245), and SR (R1752 / R1756) emerged as top predictors for predicting leaf water potential, significantly contributing to the highest model performance. The base learner models achieved an R-squared value of approximately 0.81, indicating their capacity to explain 81% of the variance. Research is underway to develop a neural vegetation index (NVI) that automates the process of index development by searching for specific wavelengths in the space ratio of linear functions of reflectance. The NVI framework could work across species and predict different physiological parameters.

Keywords: hyperspectral reflectance, water potential, spectral indices, machine learning, wild blueberries, optimal bands

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Authors: Makhosi Buthelezi

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Mitochondrial DNA cytochrome c oxidase I (COI) gene analyses linked the South African groundnut leaf miner (GLM) to the Australian soya bean moth Aproaerema simplexella (Walker) and Indian Aproaerema modicella (Deventer). Thus, the genetic relatedness of GLM, A. simplexela, and A. modicella was examined by performing mitochondrial and nuclear (COI, cytochrome oxidase subunit II (COII), mitochondrial cytochrome b (CYTB), nuclear ribosomal 28S (28S) and intergenic spacer elongation factor-1 alpha ( EF-1 ALPHA) on 44 specimens collected from South Africa, four from Mozambique, and three each from single locations in India and Australia. Phylogenetic analyses were conducted using the Maximum Parsimony (MP) and Neighbour-Joining (NJ) methods. All of the datasets of the five DNA gene regions that were sequenced were also analyzed using the Basic Local Alignment Search Tool (BLAST) to find the closest matches for inclusion in the phylogenetic trees as outgroups and for purposes of information. In the phylogenetic trees for COI, COII, cytb and EF-1 ALPHA, a similar pattern was observed in the way that the sequences assembled into different groups; i.e., some sequences of A. simplexella from Australia were grouped separately from the others, but some Australian sequences grouped with those of the GLM from South Africa, India, and Mozambique. In the phylogenetic tree for 28S, all sequences from South Africa, Australia, India, and Mozambique grouped together and formed one group. For COI, genetic pairwise distance ranged from 0.97 to 3.60 %, for COII it ranged from 0.19% to 2.32%, for cytb it ranged from 0.25 to 9.77% and for EF-1 ALPHA it ranged 0.48 to 6.99%. Results of this study indicate that these populations are genetically related and presumably constitute a single species. Thus, further molecular and morphological studies need to be undertaken in order to resolve this apparent conundrum on the taxonomy of these populations.

Keywords: aproaerema modicella, aproaerema simplexella, mitochondrial DNA, nuclear DNA

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Authors: Saranya Ganapathy, Megha N. Parajulee, Michael San Francisco, Hong Zhang

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Insect pests are a serious threat to agricultural productivity. Use of chemical pesticides, the predominant control method thus far, has resulted in environmental damage, pest resurgence, and negative effects on non-target species. Genetically modified (GM) crops offer a promising alternative, and Bacillus thuringiensis endotoxin genes have played a major role in this respect. However, to overcome insect tolerance issues and to broaden the target range, it is critical to identify alternative-insecticidal toxins working through novel mechanisms. Our research group has identified a kinase from Chilo iridescent virus (CIV; Family Iridoviridae) that has insecticidal activity and designated it as ISTK (Iridovirus Serine/Threonine Kinase). A 35 kDa truncated form of ISTK, designated iridoptin, was obtained during expression and purification of ISTK in the yeast system. This yeast-expressed CIV toxin induced 50% mortality in cotton aphids and 100% mortality in green peach aphids (GPA). Optimized viral genes (o-ISTK and o-IRI) were stably transformed into the model plant, Arabidopsis. PCR analysis of genomic DNA confirmed the presence of the gene insert (oISTK/oIRI) in selected transgenic lines. The further screening was performed to identify the PCR positive lines that showed expression of respective toxins at the polypeptide level using Western blot analysis. The stable lines expressing either of these two toxins induced moderate to very high mortality in GPAs and significantly affected GPA development and fecundity. The aphicidal potential of these transgenic Arabidopsis lines will be presented.

Keywords: Chilo iridescent virus, insecticidal toxin, iridoviruses, plant-incorporated protectants, serine/threonine kinase

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Authors: Checkfaith I. Aizebeoje, Temitope O. Lawal, Bolanle A. Adeniyi

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The overuse and abuse of antibiotics in treating zoonotic infections in humans and opportunistic infections in rabbit has contributed to the increase in antimicrobial drug resistance, therefore, an alternative to antibiotics is needed in treating these infections. The study was carried out to determine the antimicrobial activity of lactic acid bacteria (LAB) isolated from rabbit’s faeces against multidrug-resistant (MDR) pathogens isolated from the same rabbit. Twelve faecal samples and twelve swabs from fur samples were randomly collected aseptically from apparently healthy rabbits from Ajibode, Ibadan and University of Ibadan research farm in Ibadan, Oyo state, Nigeria. Lactic acid bacteria and multidrug-resistant pathogens were isolated using appropriate agar media and identified by partial sequencing of the 16SrRNA gene. Antibiotic susceptibility pattern of isolated bacteria and LAB were determined by the agar diffusion method. The antibacterial activity of the LAB against the test pathogens was determined using the agar overlay and agar diffusion methods. The pathogens Myroides gitamensis, Citrobacter rodentium, Acinetobacter johnsonii, Enterobacter oryzendophyticus and Serratia marcescens as well as twenty-eight (28) species of LAB belonging to Acetobacter and Lactobacillus genera were identified and characterized. Lactobacillus plantarum had the highest (60.71%) occurrence of the LAB. Viable cells and cell free supernatant (CFS) of isolated LAB inhibited the growth of the test organisms with the largest zone of inhibition (40 mm) produced by Lactobacillus plantarum against Citrobacter rodentium. This study showed that LAB from rabbit possess considerable antibacterial activity against multidrug-resistant bacteria from the same environment.

Keywords: antibacterial activities, cell-free supernatant, lactic acid bacteria; multidrug-resistant pathogens, rabbits’ faeces

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Authors: Hussein Bizimana, Osman Sönmez

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Constructed wetlands are current the most widely recognized waste water treatment option, especially in developing countries where they have the potential for improving water quality and creating valuable wildlife habitat in ecosystem with treatment requirement relatively simple for operation and maintenance cost. Lack of grey waste water treatment facilities in Kigali İnstitute of Science and Technology in Rwanda, causes pollution in the surrounding localities of Rugunga sector, where already a problem of poor sanitation is found. In order to treat grey water produced at Kigali İnstitute of Science and Technology, with high BOD concentration, high nutrients concentration and high alkalinity; a Horizontal Sub-surface Flow pilot-scale constructed wetland was designed and can operate in Kigali İnstitute of Science and Technology. The study was carried out in a sedimentation tank of 5.5 m x 1.42 m x 1.2 m deep and a Horizontal Sub-surface constructed wetland of 4.5 m x 2.5 m x 1.42 m deep. The grey waste water flow rate of 2.5 m3/d flew through vegetated wetland and sandy pilot plant. The filter media consisted of 0.6 to 2 mm of coarse sand, 0.00003472 m/s of hydraulic conductivity and cattails (Typha latifolia spp) were used as plants species. The effluent flow rate of the plant is designed to be 1.5 m3/ day and the retention time will be 24 hrs. 72% to 79% of BOD, COD, and TSS removals are estimated to be achieved, while the nutrients (Nitrogen and Phosphate) removal is estimated to be in the range of 34% to 53%. Every effluent characteristic will meet exactly the Rwanda Utility Regulatory Agency guidelines primarily because the retention time allowed is enough to make the reduction of contaminants within effluent raw waste water. Treated water reuse system was developed where water will be used in the campus irrigation system again.

Keywords: constructed wetlands, hydraulic conductivity, grey waste water, cattails

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Authors: Dukali Abujnah

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This study was undertaken to knowledge the chemical composition of some farm animals (cows ) by using ultrasonic milk analyzer. This method was used first time in our country in the field of milk hygiene to determine and comparison the composition percent of raw milk for farm animals as attempt to protect the health of consumers 240 raw milk samples for each species of animals included in this study for two different season of year first (Novembe 2014 - January 2015) , second from (May – Novembe2015) .The chemical analysis. In this way, a range of building suitable to put in the Libyan market product structures, and milk, for use as a new product or exposure to treatment such as pasteurized milk and long-life may be wider study project..Controls have been implemented in operating in Libya, companies which are part of the business created by the Libyan state. It was created only after the companies that made many of the studies and research carried out on areas of Libya which it was possible to intervene. Although troops spent in recent years, companies have not yet found their balance and milk production has not yet come to those set forth in the action plans, due to problems of various types that will be discussed in this study. We believe that these data have been taken in four years if analyzed properly, can provide more accurate information to farmers in their companies want to take dairy cattle breeding. The total bacterial count was 76 and 38 cfu/ml for locally produced and imported UHT milk respectively. Coliforms were negative for all examined locally produce and imported samples. E. coli were not isolated from any of the examined UHT samples. Aerobic spore-formers count mean count was 49 and 27 cfu/ml for both examined samples respectively. Obtained results show that most of the locally produced UHT milk does not satisfy the suggested Libyan Standards for UHT milk for fat% as well as total bacterial count/ml. Economical and public health importance of the obtained results were discussed, and control measures for improving the quality of UHT locally made milk were also discussed.

Keywords: locally, UHT milks, farms animals, raw milk analysis milk

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Authors: Jonnalagadda Naveen, M. P. S. Tomar, Putluru Satish, Palanisamy Dharani

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Present investigation was carried out on a single specimen of the heart of yellow-bellied sea snake, which accidentally came to the seashore with the fisherman’s net. After the death, these specimens was preserved in 10% neutral buffered formalin and observe for its morphology. The literature cited revealed that meager information was available on the anatomy of the heart of this species of snake thus present study was planned on the gross anatomy of the heart of yellow-bellied sea snake. The heart of yellow-bellied sea snake was located between 28-35th rib in an oblique direction in the pericardial sac. It was three chambered with the complete division of atria but the ventricular cavity was incompletely divided. The apex did not show any gubernaculum cordis. The sinus venosus was the common cavity for confluence of anterior and posterior vana cava and the jugular vein was opened with anterior vena cava. The opening of posterior vena cava was slit-like and it was guarded by membranous valves whereas no valve could be observed at the opening of anterior vana cava and the jugular vein. Both the caval veins ran along the right border of the heart. Pulmonary vein was single which later divided into two branches. The length-width index for the atria was 1.33 whereas it was 1.67 for the ventricle. The atrioventricular canal was situated slightly towards the left of the midline of the heart and was divided into a right cavum pulmonale and left cavum arteriosum of which the right one was slightly larger and longer than the left. The cavum venosum was present in between the cavum pulmonale and the cavum arteriosum. The Ventricle was elongated triangle muscular compartment with ventrally located apex. Internally the cavity of ventricle was divided into two partial chambers dorsally by a muscular ridge and ventrally by an incomplete inter ventricular septum.

Keywords: aorta, atrium, heart, sea snake, sinus venosus, ventricle

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Authors: Azadeh Yasari, Michael Ganzle, Stephen Strelkov, Nuanyi Liang, Jonathan Curtis, Nat N. V. Kav

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Pathogenic fungi cause significant yield losses and quality reductions to major crops including wheat, canola, and barley. Toxic metabolites produced by phytopathogenic fungi also pose significant risks to animal and human health. Extensive application of synthetic fungicides is not a sustainable solution since it poses risks to human, animal and environmental health. Unsaturated fatty acids may provide an environmentally friendly alternative because of their direct antifungal activity against phytopathogens as well as through the stimulation of plant defense pathways. The present study assessed the in vitro and in vivo efficacy of two hydroxy fatty acids, coriolic acid and ricinoleic acid, against the phytopathogens Fusarium graminearum, Pyrenophora tritici-repentis, Pyrenophora teres f. teres, Sclerotinia sclerotiorum, and Leptosphaeria maculans. Antifungal activity of coriolic acid and ricinoleic acid was evaluated using broth micro-dilution method to determine the minimum inhibitory concentration (MIC). Results indicated that both ricinoleic acid and coriolic acid showed antifungal activity against phytopathogens, with the strongest inhibitory activity against L. maculans, but the MIC varied greatly between species. An antifungal effect was observed for coriolic acid in vivo against pathogenic fungi of wheat and barley. This effect was not correlated to the in vitro activity because ricinoleic acid with equivalent in vitro antifungal activity showed no protective effect in vivo. Moreover, neither coriolic acid nor ricinoleic acid controlled fungal pathogens of canola. In conclusion, coriolic acid inhibits some phytopathogens in vivo and may have the potential to be an effective crop protection agent.

Keywords: coriolic acid, minimum inhibitory concentration, pathogenic fungi, ricinoleic acid

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Authors: G. Quiroga-Cubides, M. Cruz, E. Grijalba, J. Sanabria, A. Ceballos, L. García, M. Gómez

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Beauveria sp. has been used as an entomopathogenic microorganism for biological control of various plant pests such as whitefly, thrips, aphids and chrysomelidaes (including Cerotoma tingomariana species), which affect soybean crops in Colombia´s Altillanura region. Therefore, a biopesticide prototype based on B. bassiana strain Bv060 was developed at Corpoica laboratories. For the production of B. bassiana conidia, a baseline fermentation was performed at laboratory in a solid medium using broken rice as a substrate, a temperature of 25±2 °C and a relative humidity of 60±10%. The experimental design was completely randomized, with a three-time repetition. These culture conditions resulted in an average conidial concentration of 1.48x10^10 conidia/g, a yield of 13.07 g/kg dry substrate and a productivity of 8.83x10^7 conidia/g*h were achieved. Consequently, the objective of this study was to evaluate the influence of the particle size reduction of rice (<1 mm) and the addition of a complex nitrogen source over conidia production and efficiency parameters in a solid-state fermentation, in a completely randomized experiment with a three-time repetition. For this aim, baseline fermentation conditions of temperature and humidity were employed in a semisolid culture medium with powdered rice (10%) and a complex nitrogen source (8%). As a result, it was possible to increase conidial concentration until 9.87x10^10 conidia/g, yield to 87.07 g/g dry substrate and productivity to 3.43x10^8 conidia/g*h. This suggested that conidial concentration and yield in semisolid fermentation increased almost 7 times compared with baseline while the productivity increased 4 times. Finally, the designed system for semisolid-state fermentation allowed to achieve an easy conidia recovery, which means reduction in time and costs of the production process.

Keywords: Beauveria bassiana, biopesticide, solid state fermentation, semisolid medium culture

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Authors: Maria C. Proença, Maria T. Rebelo, Maria J. Alves, Sofia Cunha

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Vector-borne diseases are transmitted to humans by mosquitos, sandflies, bugs, ticks, and other vectors. Some are re-transmitted between vectors, if the infected human has a new contact when his levels of infection are high. The vector is infected for lifetime and can transmit infectious diseases not only between humans but also from animals to humans. Some vector borne diseases are very disabling and globally account for more than one million deaths worldwide. The mosquitoes from the complex Culex pipiens sl. are the most abundant in Portugal, and we dispose in this moment of a data set from the surveillance program that has been carried on since 2006 across the country. All mosquitos’ species are included, but the large coverage of Culex pipiens sl. and its importance for public health make this vector an interesting candidate to assess risk of disease amplification. This work focus on ports and airports identified as key areas of high density of vectors. Mosquitoes being ectothermic organisms, the main factor for vector survival and pathogen development is temperature. Minima and maxima local air temperatures for each area of interest are averaged by month from data gathered on a daily basis at the national network of meteorological stations, and interpolated in a geographic information system (GIS). The range of temperatures ideal for several pathogens are known and this work shows how to use it with the meteorological data in each port and airport facility, to focus an efficient implementation of countermeasures and reduce simultaneously risk transmission and mitigation costs. The results show an increased alert with decreasing latitude, which corresponds to higher minimum and maximum temperatures and a lower amplitude range of the daily temperature.

Keywords: human health, risk assessment, risk management, vector-borne diseases

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Authors: Adane Gebresilassie Hailemariam, Belete Yilma Hirpaye

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The enhanced concentrations of heavy metals, especially in sediments, may indicate human-induced perturbations rather than natural enrichment through geological weathering. Heavy metals are non-biodegradable, persist in the environment, and are concentrated up to the food chain, leading to enhanced levels in the liver and muscle tissues of fishes, aquatic bryophytes, and aquatic biota. Marine organisms, in general fish in particular, accumulate metals to concentrations many times higher than present in water or sediment as they can take up metals in their organs and concentrate at different levels. Thus, metals acquired through the food chain due to pollution are potential chemical hazards, threatening consumers. The Nile tilapia (oreochromic niloticus), catfish (clarius garpinus), and water samples were collected from five sampling sites, namely, inlet-1, inlet-2, center, outlet-1 and outlet-2 of Lake Chamo. The concentration of major and trace metals Na, K, Mg, Ca, Cr, Co, Ni, Mn and Cu in the two fish muscles, gill and liver, was determined using an atomic absorption spectrometer (AAS) and flame photometer (FP). Metal concentrations in the water have also been evaluated within the two consecutive seasons, winter (dry) and spring (wet). The results revealed that the concentration of those metals in Tilapia’s (O. niloticus) muscle, gill, and liver were Na 44.5, 35.1, 28, Mg 2.8, 8.41, 4.61, K 43, 32, 30, Ca 1.5, 6.0, 5.5, Cr 0.91, 1.2, 3.5, Co 3.0, 2.89, 2.62, Ni 0.94, 1.99, 2.2, Mn 1.23, 1.51, 1.6 and Cu 1.1, 1.99, 3.5 mg kg-1 respectively and in catfish’s muscle, gill and liver Na 25, 39, 41.5, Mg 4.8, 2.87, 6, K 29, 38, 40, Ca 2.5, 8.10, 3.0, Cr 0.65, 3.5, 5.0, Co 2.62, 1.86, 1.73, Ni 1.10, 2.3, 3.1, Mn 1.54, 1.57, 1.59 and Cu 1.01, 1.10, 3.70 mg kg-1 respectively. The highest accumulation of Na and K were observed for tilapia muscle and catfish gill, Mg and Ca got higher in tilapia gill and catfish liver, while Co is higher in muscle of the two fish. The Cr, Ni, Mn and Cu levels were higher in the livers of the two fish species. In conculusion, metal toxicity through food chain is the current dangerous issue for human and othe animals. This needs deep focus to promot the health of living animals. The Details of the work are going to be discussed at the conference.

Keywords: bioaccumulation, catfish, essential metals, nile tilapia

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Authors: A. Ribeiro, N. Valério, C. Vilarinho, J. Araujo, J. Carvalho

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Res2ValHUM project involves institutions from the Spanish Autonomous Region of Galicia and the north of Portugal (districts of Porto and Braga) and has as overall objectives of promotion of composting as an process for the correct managing of organic waste, valorization of compost in different fields or applications for the constitution of products with high added value, reducing of raw materials losses, and reduction of the amount of waste throw in landfills. Three main actions were designed to achieve the objectives: development of a management tool to improve collection and residue channeling for composting, sensibilization of the population for composting and characterization of the chemical and biological properties of compost and humic and fulvic substances to envisage high-value applications of compost. Here we present the cooperative activity of Galician and northern Portuguese institutions to valorize organic waste in both regions with common socio-economic characteristics and residue management problems. Results from the creation of the resource manage tool proved the existence of a large number of agricultural wastes that could be valorized. In the North of Portugal, the wastes from maize, oats, potato, apple, grape pomace, rye, and olive pomace can be highlighted. In the Autonomous Region of Galicia the wastes from maize, wheat, potato, apple, and chestnuts can be emphasized. Regarding the isolation and identification of microbial consortia from compost samples, results proved microorganisms belong mainly to the genus Bacillus spp. Among all the species identified in compost samples, Bacillus licheniformis can be highlighted in the production of humic and fulvic acids.

Keywords: agricultural wastes, Bacillus licheniformis, Bacillus spp., humic-acids, fulvic-acids

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Authors: Essam A. Makky, Chan Cai Wen, Muna Jalal, Mashitah M. Yusoff

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Termites are considered as important pests that could cause severe wood damage and economic losses in urban, agriculture and forest of Malaysia. The ability of termites to degrade cellulose depends on association of gut cellulolytic microflora or better known as mutual symbionts. With the idea of disrupting the mutual symbiotic association, better pest control practices can be attained. This study is aimed to isolate cellulolytic bacteria from the gut of termites and carry out antibacterial studies for the termite. Confirmation of cellulase activity is done by qualitative and quantitative methods. Impacts of antibiotics and their combinations, as well as heavy metals and disinfectants, are conducted by using disc diffusion method. Effective antibacterial agents are then subjected for termite treatment to study the effectiveness of the agents as termiticides. 24 cellulolytic bacteria are isolated, purified and screened from the gut of termites. All isolates were identified as Gram-negative with either rod or cocci in shape. For antibacterial studies result, isolates were found to be 100% sensitive to 4 antibiotics (rifampicin, tetracycline, gentamycin, and neomycin), 2 heavy metals (cadmium and mercury) and 3 disinfectants (lactic acid, formalin, and hydrogen peroxide). 22 out of 36 antibiotic combinations showed synergistic effect while 15 antibiotic combinations showed an antagonistic effect on isolates. The 2 heavy metals and 3 disinfectants that showed 100% effectiveness, as well as 22 antibiotic combinations, that showed synergistic effect were used for termite control. Among the 27 selected antibacterial agents, 12 of them were found to be effective to kill all the termites within 1 to 6 days. Mercury, lactic acid, formalin and hydrogen peroxide were found to be the most effective termiticides in which all termites were killed within 1 day only. These effective antibacterial agents possess a great potential to be a new application to control the termite pest species in the future.

Keywords: antibacterial, cellulase, termicide, termites

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Authors: Koyar Rane

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Nano- to submicron size particles of narrow particle size distribution of semi-conducting TiO₂, ZnO, NiO, CuO, Fe₂O₃ have been synthesized by novel hydrazine method and tested for their gas sensing, photocatalytic and bactericidal activities and the behavior found to be enhanced when the oxides in the thin film forms, that obtained in a specially built spray pyrolysis reactor. Hydrazine method is novel in the sense, say, the UV absorption edge of the white pigment grade wide band gap (~3.2eV) TiO₂ and ZnO shifted to the visible region turning into yellowish particles, indicating modification occurring the band structure. The absorption in the visible region makes these oxides visible light sensitive photocatalysis in degrading pollutants, especially the organic dyes which otherwise increase the chemical oxygen demand of the drinking water, enabling the process feasible not under the harsh energetic UV radiation regime. The electromagnetic radiations on irradiation produce electron-hole pairs Semiconductor + hν → e⁻ + h⁺ The electron-hole pairs thus produced form Reactive Oxygen Species, ROS, on the surface of the semiconductors, O₂(adsorbed)+e⁻ → O₂• - superoxide ion OH-(surface)+h⁺ →•OH - Hydroxyl radical The ROS attack the organic material and micro-organisms. Our antibacterial studies indicate the metal oxides control the Biological Oxygen Demand (BOD) of drinking water which had beyond the safe level normally found in the municipal supply. Metal oxides in the thin film form show overall enhanced properties and the films are reusable. The results of the photodegradation and antibactericidal studies are discussed. Gas sensing studies too have been done to find the versatility of the multifunctional metal oxides.

Keywords: hydrazine method, visible light sensitive, photo-degradation of dyes, water/airborne pollutant

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Authors: Nadine Wiesmann, Melanie Viel, Christoph Buhr, Rachel Tanner, Wolfgang Tremel, Juergen Brieger

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Recidivation of tumors and the development of resistances against the classical anti-tumor approaches represent a major challenge we face when treating cancer. In order to master this challenge, we are in desperate need of new treatment options beyond the beaten tracks. Zinc oxide nanoparticles (ZnO NPs) represent such an innovative approach. Zinc oxide is characterized by a high level of biocompatibility, concurrently ZnO NPs are able to exert anti-tumor effects. By concentration of the nanoparticles at the tumor site, tumor cells can specifically be exposed to the nanoparticles while low zinc concentrations at off-target sites are tolerated well and can be excreted easily. We evaluated the toxicity of ZnO NPs in vitro with the help of immortalized tumor cell lines and primary cells stemming from healthy tissue. Additionally, the Chorioallantoic Membrane Assay (CAM Assay) was employed to gain insights into the in vivo behavior of the nanoparticles. We could show that ZnO NPs interact with tumor cells as nanoparticulate matter. Furthermore, the extensive release of zinc ions from the nanoparticles nearby and within the tumor cells results in overload with zinc. Beyond that, ZnO NPs were found to further the generation of reactive oxygen species (ROS). We were able to show that tumor cells were more prone to the toxic effects of ZnO NPs at intermediate concentrations compared to fibroblasts. With the help of ZnO NPs covered by a silica shell in which FITC dye was incorporated, we were able to track ZnO NPs within tumor cells as well as within a whole organism in the CAM assay after injection into the bloodstream. Depending on the applied concentrations, selective tumor cell killing seems feasible. Furthermore, the combinational treatment of tumor cells with radiotherapy and ZnO NPs shows promising results. Still, further investigations are needed to gain a better understanding of the interaction between ZnO NPs and the human body to be able to pave the way for their application as an innovative anti-tumor agent in the clinics.

Keywords: metal oxide nanoparticles, nanomedicine, overcome resistances against classical treatment options, zinc oxide nanoparticles

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Authors: Mohamad Mohsen Homayouni, Niloofar Taghipour, Ahmad Reza Memar, Niloofar Khalaji, Hamed Kiani, Seyyed Javad Seyyed Tabaei

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Background and Objective: Cryptosporidium is a coccidian protozoan parasite; its oocysts in surface water are a global health problem. Due to the low number of parasites in the water resources and the lack of laboratory culture, rapid and sensitive method for detection of the organism in the water resources is necessarily required. We applied modified acid-fast staining and PCR for the detection of the Cryptosporidium spp. and analysed the genotypes in 55 samples collected from surface water. Methods: Over a period of nine months, 55 surface water samples were collected from the five rivers in Tehran, Iran. The samples were filtered by using cellulose acetate membrane filters. By acid fast method, initial identification of Cryptosporidium oocyst were carried out on surface water samples. Then, nested PCR assay was designed for the specific amplification and analysed the genotypes. Results: Modified Ziehl-Neelsen method revealed 5–20 Cryptosporidium oocysts detected per 10 Liter. Five out of the 55 (9.09%) surface water samples were found positive for Cryptosporidium spp. by Ziehl-Neelsen test and seven (12.7%) were found positive by nested PCR. The staining results were consistent with PCR. Seven Cryptosporidium PCR products were successfully sequenced and five gp60 subtypes were detected. Our finding of gp60 gene revealed that all of the positive isolates were Cryptosporidium parvum and belonged to subtype families IIa and IId. Conclusion: Our investigations were showed that collection of water samples were contaminated by Cryptosporidium, with potential hazards for the significant health problem. This study provides the first report on detection and genotyping of Cryptosporidium species from surface water samples in Iran, and its result confirmed the low clinical incidence of this parasite on the community.

Keywords: Cryptosporidium spp., membrane filtration, subtype, surface water, Iran

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Authors: Marlen Acosta Alamo, Lisa Manne, Richard Veit

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Habitat loss due to land use change is one of the leading causes of biodiversity loss worldwide. This form of habitat loss is a non-random phenomenon since the same environmental factors that make an area suitable for supporting high local biodiversity overlap with those that make it attractive for urban development. We aimed to compare the effect of two non-random habitat loss predictors on the richness, abundance, and rarity of nature-affiliated and human-affiliated North American breeding birds. For each group of birds, we simulated the non-random habitat loss using two predictors: the House Price Index as a measure of the attractiveness of an area for humans and the Normalized Difference Vegetation Index as a proxy for primary productivity. We compared the results of the two non-random simulation sets and one set of random habitat loss simulations using an analysis of variance and followed up with a Tukey-Kramer test when appropriate. The attractiveness of an area for humans predicted estimates of richness loss and increase of rarity higher than primary productivity and random habitat loss for nature-affiliated and human-affiliated birds. For example, at 50% of habitat loss, the attractiveness of an area for humans produced estimates of richness at least 5% lower and of a rarity at least 40% higher than primary productivity and random habitat loss for both groups of birds. Only for the species abundance of nature-affiliated birds, the attractiveness of an area for humans did not outperform primary productivity as a predictor of biodiversity following habitat loss. We demonstrated the value of the House Price Index, which can be used in conservation assessments as an index of the risks of habitat loss for natural communities. Thus, our results have relevant implications for sustainable urban land-use planning practices and can guide stakeholders and developers in their efforts to conserve local biodiversity.

Keywords: biodiversity loss, bird biodiversity, house price index, non-random habitat loss

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Authors: Mohammadjavad Mehrabanpour, Maryam Ranjbar Bushehri, Dorsa Mehrabanpour

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Respiratory diseases are the most important problems in the country’s poultry industry, particularly when it comes to broiler flocks. Ornithobacterium rhinotracheale (ORT) is a species that causes poor performance in growth rate, egg production, and mortality. This pathogen causes a respiratory infection including pulmonary alveolar inflammation, and pneumonia of birds throughout the world. Newcastle disease (ND) is a highly contagious disease in poultry, and also, it causes considerable losses to the poultry industry. The aim of this study was to evaluate the simultaneous occurrence of ORT and ND and NDV isolation by inoculation in embryonated eggs and confirmed by RT-PCR in broiler chicken flocks in Fars province. In this study, 318 blood and 85 tissue samples (brain, trachea, liver, and cecal tonsils) were collected from 15 broiler chicken farms. Survey serum antibody titers against ORT by using a commercial enzyme-linked immunosorbent assay (ELISA) kit performed. Evaluation of antibody titer against ND virus is performed by hemagglutination inhibition test. Virus isolation with chick embryo eggs 9-11 and RT-PCR method were carried out. A total of 318 serum samples, 135 samples (42.5%) were positive for antibodies to ORT and titer of HI antibodies against NDV in 122 serum samples (38/4%) were 7-10 (log2) and 61 serum samples (19/2%) had occurrence antibody titer against Newcastle virus and ORT. Results of the present study indicated that 20 tissue samples were positive in embryonated egg and in rapid hemagglutination (HA) test. HI test with specific ND positive serum confirmed that 6 of 20 samples. PCR confirmed that all six samples were positive and PCR products of samples indicated 535-base pair fragments in electrophrosis. Due to the great economic importance of these two diseases in the poultry industry, it is necessary to design and implement a comprehensive plan for prevention and control of these diseases.

Keywords: ELISA, Ornithobacterium rhinotracheale, newcastle disease, seroprevalence

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Authors: Odunayo Christy Atewolara-Odule, Olapeju O. Aiyelaagbe

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The search for new natural anti-oxidants has grown tremendously over the years because reactive oxygen species (ROS) production and oxidative stress have been linked to a large number of human degenerative diseases, such as cancer, cardiovascular diseases, inflammation, and diabetes. Tapinanthus bangwensis, a parasitic plant commonly known as mistletoe belonging to the Loranthaceae family, is mostly employed traditionally to treat inflammation, cancer, diabetes, and hypertension to mention a few. In this study, air-dried pulverized leaves and stem of Tapinanthus bangwensis were successively extracted with n-hexane, ethyl acetate, and methanol to give the corresponding crude extracts. The extracts were purified by column chromatography and high-performance liquid chromatography to give the isolated compounds. Structural elucidation was done using mass spectrometry, Fourier transform infra-red, 1D and 2D NMR spectroscopy. The antioxidant activity of the compounds was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ascorbic acid as standard. Three compounds; Friedelin, Eudesmic acid (3,4,5-trimethoxybenzoic) and Methyl-3,4,5-trimethoxybenzoate were isolated from the extracts of Tapinanthus bangwensis. Friedelin was isolated from the ethyl acetate extract of the stem while the two other compounds were isolated from the methanol extract of the leaves. The percentages of free radical scavenging activities of the compounds are as follows: Friedelin, 73.69%, methyl-3,4,5-trimethoxybenzoate, 79.33% and eudesmic, 87.68% anti-oxidant activity which were quite comparable to 93.96% given by ascorbic acid. We are reporting, to our best knowledge, for the first time the occurrence of friedelin and eudesmic acid in Tapinanthus bangwensis. The high anti-oxidant activity of these compounds supports the use of this plant in the management of diabetes and hypertension as they will be useful in combating complications arising from the disease.

Keywords: column chromatography, eudesmic acid, friedelin, Tapinanthus bangwensis

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Authors: Amit Kaushik

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Equus kiang kiang is an indigenous large-bodied herbivore species, and in India, it is restricted to limited geographies of Ladakh. One of such areas is the Tsokar Basin. With the rise in global pashmina demand, the livestock numbers have grown significantly. Previous studies have reported conflict between a nomadic pastoral community, the Changpas, and kiang. Absentee pastoralism (in lieu of pure pastoralism) and tourism are two major economic activities among the local people. However, the social, economic, political, and ecological changes are inevitable in such a contemporary system. The study examines several factors influencing the local pastoral economy and focuses on the presence of two non-human cohabitants, kiang, and the wolf. This study used semi-structured interviews and vehicle count method in four different seasons. The results show that people perceived kiang as a threat but also reveal a level of tolerance towards them. The locals predicted high kiang numbers ranging from 200-3000 in the basin and contrastingly ranked them behind wolves, which are very few in numbers. Due to a lack of scientific pieces of evidence, the kiang population status remains obscure, and local peoples’ concerns remain unaddressed. But how this competitive dysfunctionality does take place? On one side, the rural development or the animal husbandry department aims at developing the area by providing stall-feed and tourism, whereas, on another side, the wildlife department emphasizes wildlife conservation. Therefore, the managers and planners may need to be cautious about the local socio-ecological complexities and may require inter-departmental communications. The study concludes that an interdisciplinary inquiry may be an important tool in understanding such a precarious situation and may be used in the policy-making processes.

Keywords: coexistence, human-livestock-wildlife interactions, interdisciplinary approach, kiang, policymaking, tsokar.

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Authors: Sandeep Vasaikar, Lary Obi

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Rapid and discriminative genotyping methods are useful for determining the clonality of the isolates in nosocomial or household outbreaks. Multilocus sequence typing (MLST) is a nucleotide sequence-based approach for characterising bacterial isolates. The genetic diversity and the clinical relevance of the drug-resistant Klebsiella isolates from Mthatha are largely unknown. For this reason, prospective, experimental study of the molecular epidemiology of Klebsiella isolates from patients being treated in Mthatha over a three-year period was analysed. Methodology: PCR amplification and sequencing of the drug-resistance-associated genes, and multilocus sequence typing (MLST) using 7 housekeeping genes mdh, pgi, infB, FusAR, phoE, gapA and rpoB were conducted. A total of 32 isolates were analysed. Results: The percentages of multidrug-resistant (MDR), extensively drug-resistance (XDR) and pandrug-resistant (PDR) isolates were; MDR 65.6 % (21) and XDR and PDR with 0 % each. In this study, K. pneumoniae was 19/32 (59.4 %). MLST results showed 22 sequence types (STs) were identified, which were further separated by Maximum Parsimony into 10 clonal complexes and 12 singletons. The most dominant group was Klebsiella pneumoniae with 23/32 (71.8 %) isolates, Klebsiella oxytoca as a second group with 2/32 (6.25 %) isolates, and a single (3.1 %) K. varricola as a third group while 6 isolates were of unknown sequences. Conclusions/significance: A phylogenetic analysis of the concatenated sequences of the 7 housekeeping genes showed that strains of K. pneumoniae form a distinct lineage within the genus Klebsiella, with K. oxytoca and K. varricola its nearest phylogenetic neighbours. With the analysis of 7 genes were determined 1 K. variicola, which was mistakenly identified as K. pneumoniae by phenotypic methods. Two misidentifications of K. oxytoca were found when phenotypic methods were used. No significant differences were observed between ESBL blaCTX-M, blaTEM and blaSHV groups in the distribution of Sequence types (STs) or Clonal complexes (CCs).

Keywords: phylogenetic analysis, phylogeny, klebsiella phylogenetic, klebsiella

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Authors: Aliro Villacorta, Laura Rubio, Mohamed Alaraby, Montserrat López Mesas, Victor Fuentes-Cebrian, Oscar H. Moriones, Ricard Marcos, Alba Hernández.

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Micro and nano plastics (MNPLs) are emergent environmental pollutants requiring urgent information on their potential risks to human health. One of the problems associated with the evaluation of their undesirable effects is the lack of real samples matching those resulting from the environmental degradation of plastic wastes. To such end, we propose an easy method to obtain polyethylene terephthalate nano plastics from water plastic bottles (PET-NPLs) but, in principle, applicable to any other plastic goods sources. An extensive characterization indicates that the proposed process produces uniform samples of PET-NPLs of around 100 nm, as determined by using a multi-angle and dynamic light scattering methodology. An important point to be highlighted is that to avoid the metal contamination resulting from methods using metal blades/burrs for milling, trituration, or sanding, we propose to use diamond burrs to produce metal-free samples. To visualize the toxicological profile of the produced PET-NPLs, we have evaluated their ability to be internalized by cells, their cytotoxicity, and their ability to induce oxidative stress and induce DNA damage. In this preliminary approach, we have detected their cellular uptake, but without the induction of significant biological effects. Thus, no relevant increases in toxicity, reactive oxygen species (ROS) induction, or DNA damage -as detected with the comet assay- have been observed. The use of real samples, as produced in this study, will generate relevant data in the discussion about the potential health risks associated with MNPLs exposures.

Keywords: nanoplastics, polyethylene terephthalate, physicochemical characterization, cell uptake, cytotoxicity

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Authors: Fabio T. Costa, Sergio E. Moya, Marcelo H. Sousa

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Nanocomposites designed by embedding magnetic nanoparticles into a polymeric matrix stand out as ideal magnetic-hybrid and magneto-responsive materials as sorbents for removal of pollutants in environmental applications. Covalent coupling is often desired for the immobilization of species on these nanocomposites, in order to keep them permanently bounded, not desorbing or leaching over time. Moreover, unwanted adsorbates can be separated by successive washes/magnetic separations, and it is also possible to recover the adsorbate covalently bound to the nanocomposite surface through detaching/cleavage protocols. Thus, in this work, we describe the preparation and characterization of highly-magnetizable chloromethylated polystyrene-based nanocomposite beads for selective covalent coupling in environmental applications. For synthesis optimization, acid resistant core-shelled maghemite (γ-Fe₂O₃) nanoparticles were coated with oleate molecules and directly incorporated into the organic medium during a suspension polymerization process. Moreover, the cross-linking agent ethylene glycol dimethacrylate (EGDMA) was utilized for co-polymerization with the 4-vinyl benzyl chloride (VBC) to increase the resistance of microbeads against leaching. After characterizing samples with XRD, ICP-OES, TGA, optical, SEM and TEM microscopes, a magnetic composite consisting of ~500 nm-sized cross-linked polymeric microspheres embedding ~8 nm γ-Fe₂O₃ nanoparticles was verified. This nanocomposite showed large room temperature magnetization (~24 emu/g) due to the high content in maghemite (~45 wt%) and resistance against leaching even in acidic media. Moreover, the presence of superficial chloromethyl groups, probed by FTIR and XPS spectroscopies and confirmed by an amination test can selectively adsorb molecules through the covalent coupling and be used in molecular separations as shown for the selective removal of 4-aminobenzoic acid from a mixture with benzoic acid.

Keywords: nanocomposite, magnetic nanoparticle, covalent separation, pollutant removal

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Authors: Todor Nickolov Stoyanov

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Cooperatives are present in all countries and in almost all sectors, including agriculture, forestry, food, finance, health, marketing, insurance and credit. Strong cooperatives are able to overcome many of the difficulties faced by private owners. Cooperatives use seven principles, including the 'Community Concern" principle, which enables cooperatives to work for the sustainable development of the community. The members of cooperatives may use different systems for generating year-round employment and for receiving sustainable income through performing different forestry activities. Various methods are used during the preparation of the report. These include literature reviews, statistics, secondary data and expert interviews. The members of the cooperatives are benefits exclusively from increasing the efficiency of the various products and from the overall yield of the harvest, and ultimately from achieving better profit through cooperative efforts. Cooperatives also use other types of activities that are an additional opportunity for cooperative income. There are many heterogeneous activities in the production and service sectors of the forest cooperatives under consideration. Some cooperatives serve dairies, distilleries, woodworking enterprises, tourist homes, hotels and motels, shops, ski slopes, sheep breeding, etc. Through the revenue generated by the activity, cooperatives have the opportunity to carry out various environmental and protective activities - recreation, water protection, protection of endangered and endemic species, etc., which in the case of small-scale forests cannot be achieved and the management is not sustainable. The conclusions indicate the results received in the analysis. Cooperative management of forests and forest lands gives higher incomes to individual owners. The management of forests and forest lands through cooperatives helps to carry out different environmental and protective activities. Cooperative forest management provides additional means of subsistence to the owners of poor forest lands. Cooperative management of forests and forest lands support owners to implement the forest management plans and to apply sustainable management of these territories.

Keywords: cooperative, forestry, forest owners, principles of cooperation

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Authors: Xiaodi Su, Xin Ting Zheng, Laura Sutarlie, Nur Asinah binte Mohamed Salleh, Yong Yu

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Wound healing is a dynamic process with multiple phases. Rapid profiling and quantitative characterization of inflammation and infection remain challenging. We have developed paper-like battery-free multiplexed sensors for holistic wound assessment via quantitative detection of multiple inflammation and infection markers. In one of the designs, the sensor patch consists of a wax-printed paper panel with five colorimetric sensor channels arranged in a pattern resembling a five-petaled flower (denoted as a ‘Petal’ sensor). The five sensors are for temperature, pH, trimethylamine, uric acid, and moisture. The sensor patch is sandwiched between a top transparent silicone layer and a bottom adhesive wound contact layer. In the second design, a palm-like-shaped paper strip is fabricated by a paper-cutter printer (denoted as ‘Palm’ sensor). This sensor strip carries five sensor regions connected by a stem sampling entrance that enables rapid colorimetric detection of multiple bacteria metabolites (aldehyde, lactate, moisture, trimethylamine, tryptophan) from wound exudate. For both the “\’ Petal’ and ‘Palm’ sensors, color images can be captured by a mobile phone. According to the color changes, one can quantify the concentration of the biomarkers and then determine wound healing status and identify/quantify bacterial species in infected wounds. The ‘Petal’ and ‘Palm’ sensors are validated with in-situ animal and ex-situ skin wound models, respectively. These sensors have the potential for integration with wound dressing to allow early warning of adverse events without frequent removal of the plasters. Such in-situ and early detection of non-healing condition can trigger immediate clinical intervention to facilitate wound care management.

Keywords: wound infection, colorimetric sensor, paper fluidic sensor, wound care

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Authors: P. Jost, L. Muckova, M. Matula, J. Pejchal, D. Jun, R. Stetina

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Sulfur mustard (bis(2-chlorethyl) sulfide) is highly toxic, chemical warfare agent that has been used in the past in several armed conflicts. Except for the skin, respiratory tract is one of the important routes of exposure. The elucidation and understanding of the mechanism of toxicity of SM have been effort intensive research. The multiple targets character of SM caused cellular damage resulted in activation of many different mechanisms which contribute to cellular response and participate in the final cytopathology effect. In our present work, we compared time-dependent changes in sulfur mustard exposed adult human lung fibroblasts NHLF and lung epithelial alveolar cell line A-549. Cell viability (MTT assay, Calcein-AM assay, and xCELLigence - real-time cell analysis), apoptosis (flow cytometry), mitochondrial membrane potential (Δψm, flow cytometry), reactive oxygen species induction (DC and cell cycle distribution (flow cytometry) were studied. We observed significantly decreased mitochondrial membrane potential and subsequent induction of apoptosis correlating with decreased cellular viability in the sulfur mustard exposed cells. In low concentrations, sulfur mustard-induced S-phase cell cycle arrest, on the other hand, high concentrations, cell cycle phase distribution of sulfur mustard exposed cells resembled cell cycle phase distribution of control group, which implies nonspecific cell cycle inhibition. Epithelial cells A-549 was found as more sensible to sulfur mustard toxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.

Keywords: apoptosis, cell cycle, cytotoxicity, sulfur mustard

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Authors: Benjaporn Buranrat, Nootchanat Mairuae

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Cratoxylum formosum (Jack) Dyer (CF) has been used for the traditional medicines in South East Asian and Thailand. Normally, northeast Thai vegetables have proven cytotoxic to many cancer cells. Therefore, the present study aims to explore the molecular mechanisms underlying CF-induced cancer cell death and apoptosis on breast and liver cancer cells. The cytotoxicity and antiproliferative effects of CF on the human breast MCF-7 and liver HepG2 cancer cell lines were evaluated using sulforhodamine B assay and colony formation assay. Cell migration assay was measured using wound healing assay. The apoptosis induction mechanisms were investigated through reactive oxygen species formation, caspase 3 activity, and JC-1 activity. Gene expression by real-time PCR and apoptosis related protein levels by Western blot analysis. CF induced MCF-7 and HepG2 cell death by time- and dose-dependent manner. Furthermore, CF had the greater cytotoxic potency on MCF-7 more than HepG2 cells with IC50 values of 85.70+4.52 μM and 219.03±9.96 μM respectively, at 24 h. Treatment with CF also caused a dose-dependent decrease in colony forming ability and cell migration, especially on MCF-7 cells. CF induced ROS formation, increased caspase 3 activities, and decreased the mitochondrial membrane potential, and causing apoptotic body production and DNA fragmentation. CF significantly decreased expression of the cell cycle regulatory protein RAC1 and downstream proteins, cdk6. Additionally, CF enhanced p21 and reduced cyclin D1 protein levels. CF leaf extract induced cell death, apoptosis, antimigration in both of MCF-7 and HepG2 cells. CF could be useful for developing to anticancer drug candidate for breast and liver cancer therapy.

Keywords: cratoxylum formosum (jack) dyer, breast cancer, liver cancer, cell death

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