Search results for: phragmites australis

Authors: Kleche Myriam, Ziane Nadia, Berrebbah Houria, Djebar Mohammed Reda

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The Phytoremediation has now become a necessity. Thus, in our work, we are interested in the biological wastewater treatment of Oued Meboudja. The physicochemical analysis of water after treatment showed a significant reduction of suspended matter, COD and BOD5 and rate of metals in roots for example iron and zinc. We also highlighted some significant changes in biometric and physiological parameters such as increasing the number of roots and increased respiratory metabolism through the oxygen consumption in isolated roots of Phragmites australis, placed in a polluted environment.

Keywords: phragmites australis, roots, phytoremediation, iron, zinc

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Authors: El Hadji Mamadou Sonko, Mbaye Mbéguéré, Cheikh Diop, Linda Strande

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In new approaches that are being developed for the treatment of sludge, the valorization of by-product is increasingly encouraged. In this perspective, Echinochloa pyramidalis has been successfully tested in Cameroon. Echinochloa pyramidalis is an efficient forage plant in the treatment of faecal sludge. It provides high removal rates and biosolids of high agronomic value. Thus in order to advise the use of this plant in planted drying beds in Senegal its comparison with the plants long been used in the field deserves to be carried out. That is the aim of this study showing the influence of the nature of the plants on the drainage, the purifying performances and the quality of the biosolids. Echinochloa pyramidalis, Typha australis, and Phragmites australis are the three macrophytes used in this study. The drainage properties of the beds were monitored through the frequency of clogging, the percentage of recovered leachate and the dryness of the accumulated sludge. The development of plants was followed through the measurement of the density. The purification performances were evaluated from the incoming raw sludge flows and the outflows of leachate for parameters such as Total Solids (TS), Total Suspended Solids (TSS), Total Volatile Solids (TVS), Chemical Oxygen Demand (COD), Total Kjeldahl Nitrogen (TKN), Ammonia (NH₄⁺), Nitrate (NO₃⁻), Total Phosphorus (TP), Orthophosphorus (PO₄³⁻) and Ascaris eggs. The quality of the biosolids accumulated on the beds was measured after 3 months of maturation for parameters such as dryness, C/N ratio NH₄⁺/NO₃⁻ ratio, ammonia, Ascaris eggs. The results have shown that the recovered leachate volume is about 40.4%; 45.6% and 47.3%; the dryness about 41.7%; 38.7% and 28.7%, and clogging frequencies about 6.7%; 8.2% and 14.2% on average for the beds planted with Echinochloa pyramidalis, Typha australis and Phragmites australis respectively. The plants of Echinochloa pyramidalis (198.6 plants/m²) and Phragmites australis (138 plants/m²) have higher densities than Typha australis (90.3 plants/m²). The nature of the plants has no influence on the purification performance with reduction percentages around 80% or more for all the parameters followed whatever the nature of the plants. However, the concentrations of these various leachate pollutants are above the limit values of the Senegalese standard NS 05-061 for the release into the environment. The biosolids harvested after 3 months of maturation are all mature with C/N ratios around 10 for all the macrophytes. The NH₄⁺/NO₃⁻ ratio is lower than 1 except for the biosolids originating from the Echinochloa pyramidalis beds. The ammonia is also less than 0.4 g/kg except for biosolids from Typha australis beds. Biosolids are also rich in mineral elements. Their concentrations of Ascaris eggs are higher than the WHO recommendations despite a percentage of inactivation around 80%. These biosolids must be stored for an additional time or composted. From these results, the use of Echinochloa pyramidalis as the main macrophyte can be recommended in the various drying beds planted in sub-Saharan climate conditions.

Keywords: faecal sludge, nature of plants, quality of biosolids, treatment performances

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Authors: Jin-Hong Kim

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Roles of the vegetation to mitigate the erosion of the stream bed or to facilitate the deposition of the fine sediments by the species of the aquatic plants were presented. Field investigation on the estimation of the change of the bed level and the estimation of the flow characteristics were performed. The results showed that Phragmites japonica has the mitigation function of 0.3m-0.4m of the erosion in the range of higher than 1.0m/s of flow velocity at the vegetated region. Phragmites communis has the mitigation function of 0.2m-0.3m of the erosion in the range of higher than 0.7m/s of flow velocity at the vegetated region. Salix gracilistyla has greater role than Phragmites japonica and Phragmites communis to sustain the stable channel. It has the mitigation function of 0.4m-0.5m of the erosion in the range of higher than 1.4m/s of flow velocity. Miscanthus sacchariflorus has a weak role compared with that of Phragmites japonica and Salix gracilistyla, but it has still function for sustaining the stable bed. From these results, the vegetation has effective roles to mitigate the erosion or to facilitate the deposition of the stream bed.

Keywords: aquatic plants, Phragmites japonica, Phragmites communis, Salix gracilistyla

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Authors: Christine A. Odinga, G. Sanjay, M. Mathew, S. Gupta, F. M. Swalaha, F. A. O. Otieno, F. Bux

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Wastewater discharged from municipal treatment plants contain an amalgamation of trace metals. The presence of metal pollutants in wastewater poses a huge challenge to the choice and applications of the preferred treatment method. Conventional treatment methods are inefficient in the removal of trace metals due to their design approach. This study evaluated the treatment performance of a constructed rhizofiltration system in the removal of heavy metals from municipal wastewater. The study was conducted at an eThekwni municipal wastewater treatment plant in Kingsburgh - Durban in the province of KwaZulu-Natal. The construction details of the pilot-scale rhizofiltration unit included three different layers of substrate consisting of medium stones, coarse gravel and fine sand. The system had one section planted with Phragmites australis L. and Kyllinga nemoralis L. while the other section was unplanted and acted as the control. Influent, effluent and sediment from the system were sampled and assessed for the presence of and removal of selected trace heavy metals using standard methods. Efficiency of metals removal was established by gauging the transfer of metals into leaves, roots and stem of the plants by calculations based on standard statistical packages. The Langmuir model was used to assess the heavy metal adsorption mechanisms of the plants. Heavy metals were accumulated in the entire rhizofiltration system at varying percentages of 96.69% on planted and 48.98% on control side for cadmium. Chromium was 81% and 24%, Copper was 23.4% and 1.1%, Nickel was 72% and 46.5, Lead was 63% and 31%, while Zinc was 76% and 84% on the on the water and sediment of the planted and control sides of the rhizofilter respectively. The decrease in metal adsorption efficiencies on the planted side followed the pattern of Cd>Cr>Zn>Ni>Pb>Cu and Ni>Cd>Pb>Cr>Cu>Zn on the control side. Confirmatory analysis using Electron Scanning Microscopy revealed that higher amounts of metals was deposited in the root system with values ranging from 0.015mg/kg (Cr), 0.250 (Cu), 0.030 (Pb) for P. australis, and 0.055mg/kg (Cr), 0.470mg/kg (Cu) and 0.210mg/kg,(Pb) for K. nemoralis respectively. The system was found to be efficient in removing and reducing metals from wastewater and further research is necessary to establish the immediate mechanisms that the plants display in order to achieve these reductions.

Keywords: wastewater treatment, Phragmites australis L., Kyllinga nemoralis L., heavy metals, pathogens, rhizofiltration

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Authors: Kraiem Khadija, Hamadi Kallali, Naceur Jedidi

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Constructed wetland systems are eco-technology recognized as environmentally friendly and emerging innovative solutions remediation as these systems are cost-effective and sustainable wastewater treatment systems. The performance of these biological system is affected by various factors such as plant, substrate, wastewater type, hydraulic loading rate, hydraulic retention time, water depth, and operation mood. The objective of this study was to to assess the alters of plant species on pollutants reduction and enrichment of anammox and nitrifing denitrifing bacteria in a modified vertical flow (VFCW) constructed wetland. This tests were carried out using three modified vertical constructed wetlands with a surface of 0.23 m² and depth 80 cm. It was a saturated vertical constructed wetland at the bottom. The saturation zone is maintained by the siphon structure at the outlet. The VFCW (₁) system was unplanted, VFCW (₂) planted with Typha angustofolia, and VFCW(₃) planted with Phragmites australis. The experimental units were fed with domestic wastewater and were operated by batch mode during 8 months at an average hydraulic loading rate around 20 cm day− 1. The operation cycle was two days feeding and five days rest. Results indicated that plants presence improved the removal efficiency; the removal rates of organic matter (85.1–90.9%; COD and 81.8–88.9%; BOD5), nitrogen (54.2–73%; NTK and 66–77%; NH4 -N) were higher by 10.7–30.1% compared to the unplanted vertical constructed wetland. On the other hand, the plant species had no significant effect on removal efficiency of COD, The removal of COD was similar in VFCW (₂) and VFCW (₃) (p > 0.05), attaining average removal efficiencies of 88.7% and 85.2%, respectively. Whereas it had a significant effect on NTK removal (p > 0.05), with an average removal rate of 72% versus 51% for VFCW (₂) and VFCW (₃), respectively. Among the three sets of vertical flow constructed wetlands, the VFCW(₂) removed the highest percent of total streptococcus, fecal streptococcus total coliforms, fecal coliforms, E. coli as 59, 62, 52, 63, and 58%, respectively. The presence and the plant species alters the community composition and abundance of the bacteria. The abundance of bacteria in the planted wetland was much higher than that in the unplanted one. VFCW(₃) had the highest relative abundance of nitrifying bacteria such as Nitrosospira (18%), Nitrosospira (12%), and Nitrobacter (8%). Whereas the vertical constructed wetland planted with typha had larger number of denitrifying species, with relative abundances of Aeromonas (13%), Paracoccus (11%), Thauera (7%), and Thiobacillus (6%). However, the abundance of nitrifying bacteria was very lower in this system than VFCW(₂). Interestingly, the presence of Thypha angustofolia species favored the enrichment of anammox bacteria compared to unplanted system and system planted with phragmites australis. The results showed that the middle layer had the most accumulation of anammox bacteria, which the anaerobic condition is better and the root system is moderate. Vegetation has several characteristics that make it an essential component of wetlands, but its exact effects are complex and debated.

Keywords: wastawater, constructed wetland, anammox, removal

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Authors: Ahmet Helvaci, Selcuk Dogan

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Reed in especially almost all the lakes in Western Anatolia grows naturally. Due to the abundance of reed, pyrolysis of reed is very economical and practical application. In this study, it is aimed to determine the optimum conditions for the pyrolysis of the reed which is a cheap and abundant raw material and to evaluate pyrolysis products. For this purpose, reed was used obtained from Eber Lake located in the borders of Bolvadin county of Afyonkarahisar. Optimum pyrolysis conditions have been determined by examining the effects of changes in pyrolysis temperature and pyrolysis time. The evaluation of the obtained liquid and solid pyrolysis products has been investigated. Especially evaluability of solid carbon black production of tires has been investigated. Tire samples were prepared with carbon black samples obtained as a result of the pyrolysis process at different temperatures. Then, performance tests were made and compared with reference carbon blacks, used in the market and standards. At the same time, surface area measurement analysis of carbon black samples was made and compared again with reference carbon blacks. In addition, the fuel values of liquid products were also determined by calorimeter. It has been determined that the best surface area (about 370 m²/g) for carbon black samples, for tire production is 40 minutes at 500ᵒC. It was also found that the best result for evaluation studies in tire production was carbon black samples obtained at 450ᵒC pyrolysis temperature. In addition, it was seen that the calorimetry results of the liquid product obtained during 60 minutes of pyrolysis were quite good (around 5500 kcal/kg).

Keywords: evaluation of products, optimization, pyrolysis, reed

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Authors: M. R. Haji Hajikolaei, A. A. Nikvand, A. R. Ghadrdan, M. Ghorbanpoor, B. Mohammadian

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This study was carried out on water buffalo for detection of Leptospira interrogans in kidney and urine and its relationship with serological findings. Blood, urine and kidney samples were taken immediately after slaughter from 353 water buffalos at Ahvaz abattoir in Khouzestan province, Iran. Sera were initially screened at serum dilution of 1:100 against seven live antigens of Leptospira interrogans: pomona, hardjo, ballum, icterohemorrhagiae, tarasovi, australis and grippotyphosa using the microscopic agglutination test (MAT) and sera with positive results were titrated against reacting antigens in serial twofold dilution from 1:100 to 1:800. The samples of kidney were embedded in paraffin wax and 5µm thick sections were stained routinely with Haematoxylin and Eosin (H&E). Polymerase chain reaction (PCR) examination was done on urine and kidney by using LipL32 gene primers. Antibodies against one or more serovars at dilution >:100 were detected in sera. The most frequent reactor was hardjo (56.2%), followed by pomona (52.3%), australis (9.8%), tarassovi (5.9%), grippotyphosa (4.5%) and icterohaemorrhagiae (3.9%). The L. interrogans were detected in 43 (12.2%) of examined buffaloes, so that 26 (8.2%) of kidney tissues, 14 (4.8%) of urine samples separately and 3 (0.84%) of both kidney and urine samples were positive in PCR. From 153 (43.3%) buffaloes with positive MAT, 24 cases were positive by PCR of kidney and/or urine samples, synchronously. Renal lesions such as interstitial nephritis, acute tubular necrosis (ATN), pyelonephritis, glomerolonephritis, renal fibrosis and hydronephrosis were found in 128 (36.3%) cases. Statistical analysis indicated that there was no significant association between results of MAT, PCR and interstitial nephritis.

Keywords: leptospiral infection, PCR, MAT, histopathology, river buffalo

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Authors: Jamal Radaideh

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Al-Asfar Lake is located about 14 km east of Al-Ahsa and is one of the most important wetland lakes in the Al Ahsa/Eastern Province of Saudi Arabia. Al-Ahsa is may be the largest oasis in the world, having an area of 20,000 hectares, in addition, it is of the largest and oldest agricultural centers in the region. The surplus farm irrigation water beside additional water supplied by treated wastewater from Al-Hofuf sewage station is collected by a drainage network and discharged into Al-Asfar Lake. The lake has good wetlands, sand dunes as well as large expanses of open and shallow water. Salt tolerant vegetation is present in some of the shallow areas around the lake, and huge stands of Phragmites reeds occur around the lake. The lake presents an important habitat for wildlife and birds, something not expected to find in a large desert. Although high evaporation rates in the range of 3250 mm are common, the water remains in the evaporation lakes during all seasons of the year is used to supply cattle with drinking water and for aquifer recharge. Investigations showed that high concentrations of nitrogen (N), phosphorus (P), biological oxygen demand (BOD), chemical oxygen demand (COD) and salinity discharge to Al Asfar Lake from the D2 drain exist. It is expected that the majority of BOD, COD and N originates from wastewater discharge and leachate from surplus irrigation water which also contribute to the majority of P and salinity. The significant content of nutrients and biological oxygen demand reduces available oxygen in the water. The present project aimed to improve the water quality of the lake using constructed wetland trains which will be built around the lake. Phragmites reeds, which already occur around the lake, will be used.

Keywords: Al Asfar lake, constructed wetland, water quality, water treatment

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Authors: Veronica Barragan, Ligia Luna, Maria Patricia Zambrano, Carlos Bulnes, Eduardo Diaz, Talima Pearson

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Leptospirosis impacts animal production and is responsible for important economic losses in the pig industry. Infection is associated with reproductive failures that lead to abortions, stillbirth, and perinatal mortality. The leptospira serogroups that have been traditionally linked to disease in pigs are Pomona, Australis, and Tarassovi. Unfortunately, knowledge about pig leptospirosis is biased towards infection in large-scale commercial farms from developed countries, where exposure is usually limited to host-specific serotypes. The aim of our study is to describe leptospirosis in pigs from rural communities located in the coast of Ecuador-South America, where leptospirosis is endemic. A particularity of these pigs is that, because they are usually raised in the backyard of their owner’s houses, exposure to other leptospira excreted by other animals is likely to occur. Therefore, we collected 420 kidney samples from pigs sacrificed at a local slaughterhouse, and Leptospira positivity was tested in all samples by amplifying the Lipl32 gen. Our results show pathogenic Leptospira positivity in 19.3% (81/420) of pigs. Microaglutination test was performed in 60 PCR positive samples with titers >1:100 in 17 pigs, titers of 1:50 in 28 pigs, and no MAT titers in 15 pigs even though Leptospira DNA was found in their kidneys. Interestingly, reacting serovars were very diverse, with 18.3% of pig sera reacting with two or more serovars. Additionally, serovar Canicola was found in 16.7% of pigs followed by Tarassovi (10%), Australis (6.7%), Pyogenes (5%), Icterohaemorrhageae (1.7%), and Grippotyphosa (1.7%). It is also important to highlight that most of the analyzed animals came from small-scale farms where pigs may be exposed to the pathogen by exposure to other domestic and peridomestic animals such as rats, dogs, horses, donkeys, and even wildlife. This would explain the finding of non-pig adapted Leptospira serovars such as Canicola, which is commonly reported in dogs.

Keywords: Leptospira, Lipl32, peridomestic, pig, serovar

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Authors: Siti Aisya Gany, Swee Ching Tan, Sook Yee Gan

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Diminished antioxidant defense or increased production of reactive oxygen species in the biological system can result in oxidative stress which may lead to various neurodegenerative diseases including Alzheimer’s disease (AD). Microglial activation also contributes to the progression of AD by producing several pro-inflammatory cytokines, nitric oxide (NO), and prostaglandin E2 (PGE2). Oxidative stress and inflammation have been reported to be possible pathophysiological mechanisms underlying AD. In addition, the cholinergic hypothesis postulates that memory impairment in patient with AD is also associated with the deficit of cholinergic function in the brain. Although a number of drugs have been approved for the treatment of AD, most of these synthetic drugs have diverse side effects and yield relatively modest benefits. Marine algae have great potential in pharmaceutical and biomedical applications as they are valuable sources of bioactive properties such as anti-coagulation, anti-microbial, anti-oxidative, anti-cancer and anti-inflammatory. Hence, this study aimed to provide an overview of the properties of Malaysian seaweeds (Padina australis, Sargassum polycystum and Caulerpa racemosa) in inhibiting oxidative stress, neuroinflammation and cholinesterase enzymes. All tested samples significantly exhibit potent DPPH and moderate Superoxide anion radical scavenging ability (P<0.05). Hexane and methanol extracts of S. polycystum exhibited the most potent radical scavenging ability with IC50 values of 0.1572 ± 0.004 mg/ml and 0.8493 ± 0.02 for DPPH and ABTS assays, respectively. Hexane extract of C. racemosa gave the strongest superoxide radical inhibitory effect (IC50 of 0.3862± 0.01 mg/ml). Most seaweed extracts significantly inhibited the production of cytokine (IL-6, IL-1 β, TNFα) and NO in a concentration-dependent manner without causing significant cytotoxicity to the lipopolysaccharide (LPS)-stimulated microglia cells (P<0.05). All extracts suppressed cytokine and NO level by more than 80% at the concentration of 0.4mg/ml. In addition, C. racemosa and S. polycystum also showed anti-acetylcholinesterase activities with the IC50 values ranging from 0.086-0.115 mg/ml. Moreover, C. racemosa and P. australis were also found to be active against butyrylcholinesterase with IC50 values ranging from 0.118-0.287 mg/ml.

Keywords: anti-cholinesterase, anti-oxidative, neuroinflammation, seaweeds

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Authors: Álvaro Ramírez Vidal, Martín Muñoz Morales, Francisco Jesús Fernández Morales, Luis Rodríguez Romero, José Villaseñor Camacho, Javier Llanos López

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In recent times, environmental problems have led to the extensive use of biological systems to solve them. Among the different types of biological systems, the use of plants such as aquatic macrophytes in constructed wetlands and terrestrial plant species for treating polluted soils and sludge has gained importance. Though the use of constructed wetlands for wastewater treatment is a well-researched domain, the slowness of pollutant degradation and high biomass production pose some challenges. Plants used in CW participate in different mechanisms for the capture and degradation of pollutants that also can retain some pharmaceutical and personal care products (PPCPs) that are very persistent in the environment. Thus, these systems present advantages in line with the guidelines published for the transition towards friendly and ecological procedures as they are environmentally friendly systems, consume low energy, or capture atmospheric CO₂. However, the use of CW presents some drawbacks, as the slowness of pollutant degradation or the production of important amounts of plant biomass, which need to be harvested and managed periodically. Taking this opportunity in mind, it is important to highlight that this residual biomass (of lignocellulosic nature) could be used as the feedstock for the generation of carbonaceous materials using thermochemical transformations such as slow pyrolysis or hydrothermal carbonization to produce high-value biomass-derived carbons through sustainable processes as adsorbents, catalysts…, thereby improving the circular carbon economy. Thus, this work carried out the analysis of some PPCPs commonly found in urban wastewater, as salicylic acid or ibuprofen, to evaluate the remediation carried out for the Phragmites Australis. Then, after the harvesting, this biomass can be used to synthesize electrodes through hydrothermal carbonization (HTC) and produce high-value biomass-derived carbons with electrocatalytic activity to remove heavy metals and persistent pollutants, promoting circular economy concepts. To do this, it was chosen biomass derived from the natural environment in high environmental risk as the Daimiel Wetlands National Park in the center of Spain, and the rest of the biomass developed in a CW specifically designed to remove pollutants. The research emphasizes the impact of the composition of the biomass waste and the synthetic parameters applied during HTC on the electrocatalytic activity. Additionally, this parameter can be related to the physicochemical properties, as porosity, surface functionalization, conductivity, and mass transfer of the electrodes lytic inks. Data revealed that carbon materials synthesized have good surface properties (good conductivities and high specific surface area) that enhance the electro-oxidants generated and promote the removal of PPCPs and the chemical oxygen demand of polluted waters.

Keywords: constructed wetlands, carbon materials, heavy metals, pharmaceutical and personal care products, hydrothermal carbonization

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Authors: Nasreen Jeelani, Huining Shi , Di An, Lu Xia, Shuqing An

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Heavy metals contamination in aquatic ecosystem is a major environmental problem since its accumulation along the food chain pose public health risk. The concentration of heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in soil and plants species collected from different streams of Suoxu River, China was investigated. This aim was to define the level of pollutants in Suoxu River, find which plant species exhibits the greatest accumulation and to evaluate whether these species could be useful for phytoremediation. While total soil Cd, Cr, Cu, Ni, Pb, and Zn concentrations varied, respectively, from 0.09 to 0.23 , 58.6 to 98, 9.72 to 80.5, 15.3 to 41, 15.2 to 27.3 and 35 to 156 (mg-kg-1), those in plants ranged from 0.035 to 0.49, 2.91 to 75.6, 4.79 to 32.4, 1.27 to 16.1, 0.62 to10.2, 18.9 to 84.6 (mg-kg-1), respectively. Based on BCFs and TFs values, most of the studied species have potential for phytostabilization. The plants with most effective in the accumulation of metals in shoots are Phragmatis australis (TF=2.29) and Iris tectorum (TF =2.07) for Pb. While Chenopodium album, (BCF =3.55), Ranunculus sceleratus, (BCF= 3.0), Polygonum hydropiper (BCF =2.46) for Cd and Iris tectorum (BCF=2.0) for Cu was suitable for phytostabilization. Among the plant species screened for Cd, Cr, Cu, Ni, Pb and Zn, most of the species were efficient to take up more than one heavy metal in roots. Our study showed that the native plant species growing on contaminated sites may have the potential uses for phytoremediation.

Keywords: heavy metals, huaihe river catchments, sediment, plants

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Authors: Vicente A. Hernandez, Felipe Galleguillos, Rene Thibaut, Alejandro Muller

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Natural pigments have been proposed as an eco-friendly alternative to artificial pigments. Among the diverse organisms able to synthesize natural pigments, several wood colonizing fungi produce extracellular pigments which have been tested to dye fabrics at laboratory conditions with good results. However, the dyeing conditions used at laboratory level not necessary meet the real conditions in which dyeing of fabrics is conducted at industrial level. In this work, yellow and red pigments from the fungi Penicillium murcianum and Talaromyces australis, respectively, were used to dye yarn and linen fabrics using dyeing processes optimized according to the standard conditions used at industrial level. After dyeing treatments, fabrics were tested for color fastness to wash and to wet and dry rubbing, but also to tensile strength tests. Satisfactory result was obtained with both yellow and red pigments in yarn and linen, when used alone or mixed to different proportions. According to these results, natural pigments synthesized by both wood colonizing fungi have a great potential to be used in dyeing processes at industrial level.

Keywords: natural pigments, fungal pigments, yarn, linen

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Authors: Yeon Ho Je

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A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties, such as a higher insecticidal activity and improved recovery, compared to wild-type baculovirus. For the construction of NeuroBactrus, the Bacillus thuringiensis crystal protein gene (here termed cry1-5) was introduced into the Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of the polyhedrin–cry1-5–polyhedrin genes under the control of the polyhedrin promoter. In the opposite direction, an insect-specific neurotoxin gene, AaIT, from Androctonus australis was introduced under the control of an early promoter from Cotesia plutellae bracovirus by fusion of a partial fragment of orf603. The polyhedrin–Cry1-5–polyhedrin fusion protein expressed by the NeuroBactrus was not only occluded into the polyhedra, but it was also activated by treatment with trypsin, resulting in an_65-kDa active toxin. In addition, quantitative PCR revealed that the neurotoxin was expressed from the early phase of infection. NeuroBactrus showed a high level of insecticidal activity against Plutella xylostella larvae and a significant reduction in the median lethal time against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Rerecombinant mutants derived from NeuroBactrus in which AaIT and/or cry1-5 were deleted were generated by serial passages in vitro. Expression of the foreign proteins (B. thuringiensis toxin and AaIT) was continuously reduced during the serial passage of the NeuroBactrus. Moreover, polyhedra collected from S. exigua larvae infected with the serially passaged NeuroBactrus showed insecticidal activity similar to that of wild-type AcMNPV. These results suggested that NeuroBactrus could be recovered to wild-type AcMNPV through serial passaging.

Keywords: baculovirus, insecticide, neurotoxin, neurobactrus

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