Live-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions

Prion diseases are associated to the conversion of the prion protein into a misfolded pathological isoform. The mechanism of propagation of protein misfolding by protein templating remains largely unknown. Neuroblastoma cells were transfected with constructs of the prion protein fused to both CFP-GPI-anchored and to YFP-GPI-anchored and directed to its cell membrane location. Live-cell FRET imaging between the prion protein fused to CFP or YFP was measured giving consistent values of 10 ± 2%. This result was confirmed by fluorescence lifetime imaging microscopy and indicates intermolecular interactions between neighbor prion proteins. In particular, considering that a maximum FRET efficiency of 17 ± 2% was determined from a positive control consisting of a fusion CFP-YFP-GPI-anchored. A stable cell clone expressing the two fusions containing the prion protein was also selected to minimize cell-to-cell variability. In both, stable and transiently transfected cells, the FRET efficiency consistently increased in the presence of infectious prions — from 4 ± 1% to 7 ± 1% in the stable clone and from 10 ± 2% to 16 ± 1% in transiently transfected cells. These results clearly reflect an increased clustering of the prion protein on the membrane in the presence of infectious prions, which was not observed in negative control using constructs without the prion protein and upon addition of non-infected brain. Our data corroborates the recent view that the primary site for prion conversion is the cell membrane. Since our fluorescent cell clone is not susceptible to propagate infectivity, we hypothesize that the initial event of prion infectivity might be the clustering of the GPI-anchored prion protein.     

Viral safety of C1-inhibitor NF

We studied the efficacy of virus reduction by three process steps (polyethylene glycol 4000 (PEG) precipitation, pasteurization, and 15 nm virus filtration) in the manufacturing of C1-inhibitor NF. The potential prion removing capacity in this process was estimated based on data from the literature. Virus studies were performed using hepatitis A virus (HAV) and human immunodeficiency virus (HIV) as relevant viruses and bovine viral diarrhea virus (BVDV), canine parvovirus (CPV) and pseudorabies virus (PRV) as model viruses, respectively. In the PEG precipitation step, an average reduction in infectious titer of 4.5 log₁₀ was obtained for all five viruses tested. Pasteurization resulted in reduction of infectious virus of >6 log₁₀ for BVDV, HIV, and PRV; for HAV the reduction factor was limited to 2.8 log₁₀ and for CPV it was zero. Virus filtration (15 nm) reduced the infectious titer of all viruses by more than 4.5 log₁₀. The overall virus reducing capacity was >16 log₁₀ for the LE viruses. For the NLE viruses CPV and HAV, the overall virus reducing capacities were >8.7 and >10.5 log₁₀, respectively. Based on literature and theoretical assumptions, the prion reducing capacity of the C1-inhibitor NF process was estimated to be >9 log₁₀.     

Removal of TSE agents by depth or membrane filtration from plasma products

The removal of the abnormal form of prion protein i.e. PrP^SC by filtration steps in the plasma fractionation process has been investigated by immuno-Western blotting. Depth filtration has been shown to be capable of removing scrapie by 2–3 log from certain plasma product intermediates. These include cryoprecipitate supernatant, used for the manufacture of immunoglobulin and albumin, and albumin fraction V, by filtration using Pall Seitz or 3m Cuno depth filters respectively. However no significant removal occurred with immunoglobulin Fraction II after Cuno depth filtration. When 0.2 μm PVDF and Nylon membrane filters were tested, the removal of TSEs from 20% albumin was limited i.e. 0.6–1.3 log. However under protein free conditions using phosphate buffered saline, filtration was not effective in the case of a PVDF filter but very effective i.e. >2.9 log in the case of a Nylon filter.     

Optimized method for the determination of itopride in human plasma by high-performance liquid chromatography with fluorimetric detection

A high-performance liquid chromatographic method with fluorescence detection for the determination of itopride in human plasma is reported. The sample preparation was based on liquid–liquid extraction of itopride from plasma with t-butylmethylether and dichloromethane (70:30, v/v) mixture followed by a back extraction of the analyte to the phosphate buffer (pH 3.2). Liquid chromatography was performed on an octadecylsilica column (55 mm × 4 mm, 3 μm particles), the mobile phase consisted of acetonitrile–triethylamine–15 mM dihydrogenpotassium phosphate (14.5:0.5:85, v/v/v), pH of the mobile phase was adjusted to 4.8. The run time was 3 min. The fluorimetric detector was operated at 250/342 nm (excitation/emission wavelength). Naratriptan was used as the internal standard. The limit of quantitation was 9.5 ng/ml using 0.5 ml of plasma. The method precision and inaccuracy were less than 8%. The assay was applied to the analysis of samples from a bioequivalence study.     

Characterization of catechol derivative removal by lignin peroxidase in aqueous mixture

The use of lignin peroxidase (LIP) as an alternative method for the removal of four catechols (1,2-dihydroxybenzene): catechol (CAT), 4-chlorocatechol (4-CC), 4,5-dichlorocatechol (4,5-DCC) and 4-methylcatechol (4-MC) typical pollutants in wastewater derived from oil and paper industries, was evaluated. The removal of 2 mM catecholic substrates by 1 μM LIP after 1 h was in the following order: 4,5-DCC (95%) > 4-CC(90%) > CAT(55%) > 4-MC(43%). Except for 4-MC, all reactions were accompanied by the formation of insoluble products, leading to LIP precipitation. LIP was exposed to soluble or insoluble product-dependent inactivation, depending on the substrates tested, immediately at the start of the reactions. Despite immediate enzyme inactivation, removal of catecholic substrates continued, resulting in oligomeric product formation. Major oxidation products analyzed were compatible with dimeric, trimeric and tetrameric structures. Ether linkages and a benzoquinone structure were detected in two purified oligochlorocatechols.Catechol derivatives removal initiated by LIP, seems to be different for each catecholic substrate in terms of substrate consumption and transformation, and of enzyme activity.     

Sono-chemical preparation of cellulose nanocrystals from lignocellulose derived materials

This study examined the production of cellulose nanocrystals from microcrystalline wood cellulose, Avicel and recycled pulp of wood pulp using sono-chemical-assisted hydrolysis. Two hydrolysis systems: deionized water and maleic acid were evaluated. In deionized water, Avicel produced cellulose nanocrystals with average diameter of 21 ± 5 nm (minimum 15 nm and maximum 32 nm). Cellulose nanocrystals from recycled pulp were not distinctively spherical and had an average diameter of 23 ± 4 nm (minimum 14 nm and maximum 32 nm). Maleic acid (50 mM) sono-chemical assisted hydrolysis of Avicel at 15 °C and 90% power output for 9 min produced cellulose nanocrystals which were cylindrical in shape and were of dimensions, length 65 ± 19 nm and width 15 nm.     

Plasma levels of transforming growth factor-beta 1 before and after removal of low- and high-grade astrocytomas

Transforming growth factor-beta 1 (TGF-β1) has been reported to be a possible marker for a number of tumors, including brain tumors. The aim of this study was to measure the plasma levels of TGF-β1 in patients with low- and high-grade astrocytomas before and after surgery. This prospective study included 14 patients with low-grade astrocytomas and 25 with high-grade astrocytomas who underwent tumor removal and 13 controls (patients who underwent cranioplasty for skull bone defects). Plasma levels of TGF-β1 were measured in all subjects using enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curve analysis showed that when the level of TGF-β1 before tumor removal was ?2.52 ng/ml, astrocytoma was predicted with a sensitivity of 94.9% and specificity of 100%. The mean plasma level of TGF-β1 in both the low-grade and high-grade astrocytoma groups significantly decreased after tumor removal (p < 0.05); there was no significant change in TGF-β1 plasma level of the controls following surgery. Patients with high-grade astrocytomas had a significantly higher mortality rate than patients with low-grade astrocytomas (p = 0.019) and significantly shorter survival (p = 0.008). A positive correlation between TGF-β1 level after tumor removal and tumor volume was only found in the high-grade astrocytoma group (γ = 0.597, p = 0.002). The findings show that plasma TGF-β1 level was increased in patients with low-grade and high-grade astrocytoma, and that the levels significantly decreased after tumor removal in both groups. The results provide additional evidence that TGF-β1 might be useful as a tumor marker for astrocytomas.     

In-vitro effect of human cathelicidin antimicrobial peptide LL-37 on dengue virus type 2

Human Cathelicidin antimicrobial peptide LL-37 is known to have antiviral activity against many viruses. In the present study, we investigated the in-vitro effect of LL-37 on dengue virus type 2 (DENV-2) infection and replication in Vero E6 cells. To study the effect of pretreatment of virus or cells with LL-37, the virus was pretreated with different concentrations of LL-37 (2.5 μM–15 μM) or scrambled (Scr) LL-37(5 μM–15 μM) and used for infection or the cells were first treated with LL-37 and infected. To study the effect of LL-37 post infection (PI), the cells were infected first followed by addition of LL-37 to the culture medium 24 h after infection. In all conditions, after the incubation, the culture supernatant was assessed for viral RNA copy number by real time RT-PCR, infectious virus particles by focus forming unit assay (FFU) and non structural protein 1 (NS1) antigen levels by ELISA. Percentage of infection was assessed using immunoflourescence assay (IFA). The results revealed that pretreatment of virus with 10–15 μM LL-37 significantly reduced its infectivity as compared to virus control (P < 0.0001). Moreover, pretreatment of virus with 10–15 μM LL-37 significantly reduced the levels of viral genomic RNA and NS1 antigen (P < 0.0001). Treatment of virus with 10–15 μM LL-37 resulted in two to three log reduction of mean log₁₀ FFU/ml as compared to virus control (P < 0.0001). Treatment of the virus with scrambled LL-37 had no effect on percentage of infection and viral load as compared to virus control cultures (P > 0.05). Pretreatment of cells before infection or addition of LL-37 to the culture 24 h PI had no effect on viral load. Molecular docking studies revealed possible binding of LL-37 to both the units of DENV envelope (E) protein dimer. Together, the in-vitro experiments and in-silico analyses suggest that LL-37 inhibits DENV-2 at the stage of entry into the cells by binding to the E protein. The results might have implications for prophylaxis against DENV infections and need further in-vivo studies.     

Simple and rapid liquid chromatography method for determination of moxifloxacin in plasma

A high performance liquid chromatographic method for determination of moxifloxacin in human plasma was developed. The method involved deproteinisation of the sample with perchloric acid and analysis of the supernatant using a reversed-phase C₁₈ column (150 mm) and fluorescence detection at an excitation wavelength of 290 nm and an emission wavelength of 460 nm. The assay was specific for moxifloxacin and linear from 0.125 to 10.0 μg/ml. The relative standard deviation of intra- and inter-day assays was lower than 10%. The average recovery of moxifloxacin from plasma was 101%. Due to its simplicity, the assay can be used for pharmacokinetic studies of moxifloxacin.     

Anti-prion activity found in beetle grub hemolymph of Trypoxylus dichotomus septentrionalis

No remedies for prion disease have been established, and the conversion of normal to abnormal prion protein, a key event in prion disease, is still unclear. Here we found that substances in beetle grub hemolymph, after they were browned by aging for a month or heating for hours, reduced abnormal prion protein (PrP) levels in RML prion-infected cells. Active anti-prion components in the hemolymph were resistant to protease treatment and had molecular weights larger than 100 kDa. Aminoguanidine treatment of the hemolymph abolished its anti-prion activity, suggesting that Maillard reaction products are enrolled in the activity against the RML prion. However, levels of abnormal PrP in RML prion-infected cells were not decreased by incubation with the Maillard reaction products formed by amino acids or bovine serum albumin. The anti-prion components in the hemolymph modified neither cellular or cell-surface PrP levels nor lipid raft or autophagosome levels. The anti-prion activity was not observed in cells infected with 22 L prion or Fukuoka-1 prion, suggesting the anti-prion action is prion strain-dependent. Although the active components of the hemolymph need to be further evaluated, the present findings imply that certain specific chemical structures in the hemolymph, but not chemical structures common to all Maillard reaction products, are involved in RML prion formation or turnover, without modifying normal PrP expression. The anti-prion components in the hemolymph are a new tool for elucidating strain-dependent prion biology.     

Evaluation of adsorbents for separation and purification of paclitaxel from plant cell cultures

In this study, we evaluated the efficiency of different adsorbents for the removal of plant-derived impurities during the pre-purification of paclitaxel from plant cell cultures. Using the synthetic adsorbents sylopute and active clay and their major components SiO₂ and MgO, we performed adsorbent treatment and analyzed the paclitaxel precipitates recovered from hexane precipitation. When SiO₂ was used, the highest purity (～58.1%) and yield (～91.5%) of paclitaxel were obtained. We also determined differences in the effectiveness of the adsorbent treatment according to changes in the surface area, pore volume and pore diameter of SiO₂. Adsorbent treatment was more effective when pore diameter was larger (silica I [2.19 nm] < silica II [4.92 nm] < silica III [9.07 nm]). The highest purity (～74.3%) and yield (～92.9%) of paclitaxel were obtained when silica III was used in the adsorbent treatment. Pore diameter had a greater effect on the removal of plant-derived impurities during the pre-purification of paclitaxel compared with surface area and pore volume. This result could be confirmed by HPLC analysis of the absorbent after treatment and TGA of the organic substances that were bonded to the adsorbent.     

Quantification of 4-aminopyridine in plasma by capillary electrophoresis with electrokinetic injection

A rapid and sensitive CE method for the determination of 4-aminopyridine in human plasma using 3,4-diaminopyridine as an internal standard was developed and validated. The analytes were extracted from 0.5-mL aliquots of human plasma by liquid–liquid extraction, using 8 mL of ethyl ether, and injected electrokinetically into capillary electrophoresis equipment. The instrumental conditions were obtained and optimized by Design of Experiments (DOE – factorial and response surface model), having as factors: separation voltage, ionic strength (buffer concentration), pH and temperature. The response variables were migration time, resolution, tailing factor and drug peak area. After obtaining mathematically predicted values for the response variables with best factors combinations, these were reproduced experimentally in good agreement with predicted values. In addition to optimal separation conditions obtained by Design of Experiments, sensitivity was improved using electrokinetic injection at 10 kV for 10 s, and a capillary with 50 cm effective length and 100 μm I.D. The final instrumental conditions were voltage at 19 kV, capillary temperature at 15 °C, wavelength at 254 nm, and phosphate buffer 100 mM, pH 2.5 as the background electrolyte. This assay was linear over a concentration range of 2.5–80 ng/mL with a lower limit of quantification of 2.5 ng/mL. The relative standard deviation for the assay precision was <7% and the accuracy was >95%. This method was successfully applied to the quantification of 4-aminopyridine (4-AP) in plasma samples from patients with spinal cord injury.     

Efficient light absorbers based on thiophene-fused boron dipyrromethene (BODIPY) dyes

We present the development of the thiophene-fused boron dipyrromethene derivatives as efficient light absorbers. The two strategies for the evolution of the optical properties such as the peak positions of absorption wavelengths and molar extinct coefficients were established by the substituent effects: by introducing iodine groups, the bathochromic shifts of the peak positions (+15 nm) and the enhancement of molar extinct coefficients were simultaneously received owing to the heavy atom effect. Next, it was found that the modification with the trifluoromethyl group contributed to the large bathochromic shift (+60 nm) because of the lowering effect on the lowest unoccupied molecular orbital of the dye by the substituent. Finally, we obtained the dyes with large molar extinct coefficients (184,140 M^?1 cm^?1 at 592 nm, 72,180 M^?1 cm^?1 at 623 nm), sharp absorption bands, and low emissions.     

Forecasting the effect of feast and famine conditions on biological sulphate reduction in an anaerobic inverse fluidized bed reactor using artificial neural networks

The longevity and robustness of bioreactors used for wastewater treatment is determined by the activity of the microorganisms under steady and transient loading conditions. Two identical continuously operated inverse fluidized bed bioreactors (IFB), IFB R1 and IFB R2, were tested for sulphate removal under the same operating conditions for 140 d (Periods I–IV). Later, IFB R1 was used as the control reactor (Period V), while IFB R2 was operated under feast (Period V-A) and famine (Period V-B) feeding conditions for 66 d. The sulphate removal efficiency was comparable in both IFB, <20% in Period I and ∼70% during Periods II, III and IV. The robustness of the IFB was evident when the sulphate removal efficiency remained comparable during the feast Period (67 ± 15%) applied to IFB R2 compared to continuous feeding Periods (Period IV (71 ± 4%) for IFB R2 and Period V (61 ± 15%) for IFB R1). The IFB performance was modelled using a three-layered artificial neural networks (ANN) model (5-11-3) and a sensitivity analysis, the sulphate removal was found to be dependent on the COD:sulphate ratio. Besides, the robustness, resilience and adaptation time of the IFB were affected by the degree of mixing and the hydraulic retention time.     

A comparative study of surface and subsurface flow constructed wetlands for treatment of combined sewer overflows: A greenhouse experiment

The use of surface flow (SFCWs) and subsurface flow constructed wetlands (SFCWs) for the treatment of combined sewer overflows was assessed at pilot scale. Synthetic wastewater was applied in three batches with decreasing concentrations to mimic concentration profiles that are obtained in the field during overflow events. Three simulated combined sewer overflows were applied on each wetland. Composite water samples (60 in total) were taken for a period of 8 days to study the removal of total nitrogen (Ntot), NH₄–N, NO₃–N, total COD (CODtot) and total phosphorus. Redox potential, which was monitored at various locations along the wetlands, was more negative in the SSFCWs. In general, removal occurred faster in the SSFCWs and the final concentrations were lower. The removal of Ntot was only 36.6 ± 3.3% in the SFCWs due to nitrification-limiting conditions. The conditions in the SSFCWs, in contrast, seemed to promote Ntot removal (removal efficiency 96.7 ± 1.9%). The removal of P was hampered in both wetland types by reducing conditions. P that was initially removed was released again from the substrates later on. First-order removal rate constants were derived for the removal of both CODtot (SSFCWs: 1.1 ± 0.3 m d^?1; SFCWs: 0.17 ± 0.06 m d^?1) and Ntot (SSFCWs: 0.4 ± 0.1 m d^?1; SFCWs: 1.7 ± 0.5 m d^?1).     

Catalytic removal of sulfide by an immobilized sulfide-oxidase bioreactor

A bioreactor packed with chitosan immobilized sulfide-oxidase from Streptomyces species LD048 was developed to treat a liquid stream of sulfide. The inoculation system was composed of glass with a 0.7 L working volume and enzyme activity of 2 mmol S g^?1 carrier. The sulfide removal efficiency was almost 100% when the volumetric loading was increased up to 3.9 mmol S L^?1 h^?1 at a space velocity of 18 h^?1. The maximal elimination capacity was 22.1 mmol S L^?1 h^?1 with a space velocity of 72 h^?1. When the aeration was increased from 0.05 to 0.1 L min^?1, the average removal efficiency improved from 81% to 94%. A removal efficiency of 90% was obtained after 15 days of operation with a load rate of 8.9 mmol S L^?1 h^?1 and a space velocity of 14.28 h^?1. An operational equation based on the ideal plug flow bioreactor and the Michaelis–Menten model predicted the performance of this bioreactor.     

Simple liquid chromatography method for the quantification of irinotecan and SN38 in sheep plasma: Application to in vivo pharmacokinetics after pulmonary artery chemoembolization using drug eluting beads

A rapid and simple liquid chromatography–fluorescence detection (LC–FD) method was developed and validated for the simultaneous quantification of irinotecan (CPT11) and SN38 in sheep plasma. Camptothecin (CPT) was used as the internal standard. A single step protein precipitation with acetonitrile was used for sample preparation. The separation was achieved using a 5 μm C18 column (250 mm × 4.5 mm, 5 μm) with a mobile phase composed of 36 mM sodium dihydrogen phosphate dehydrate and 4 mM sodium 1 heptane sulfonate–acetonitrile (72:28), the pH of the mobile phase was adjusted to 3. The flow rate was 1.45 mL/min and the fluorescence detection was operated at 355/515 nm (excitation/emission wavelengths). The run time was 13 min. The method was validated with respect to selectivity, extraction recovery, linearity, intra- and inter-day precision and accuracy, limit of quantification and stability. The method has a limit of quantification of 5 ng/mL for both CPT11 and SN38. The assay was linear over concentrations ranging from 5 to 5000 ng/mL and to 240 ng/mL for CPT11 and SN38, respectively. This method was used successfully to perform plasma pharmacokinetic studies of CPT11 after pulmonary artery embolization (PACE) in a sheep model. It was also validated for CPT11 and SN38 analysis in sheep lymph and human plasma.     

Social ranking and plasma progesterone levels in goats

Progesterone is essential for maintaining pregnancy in goats, and embryonic losses may be a consequence of the reduction in circulating progesterone levels close to the time of implantation. Some evidence exists regarding social dominance affecting the plasma progesterone levels in several species—where dominant females conceive earlier. The objective of this research was to determine whether serum progesterone levels differ in goats of different social status. A behavioural study was conducted for 10 days in a herd of 57 does and an individual success index (SI) was calculated according to the result of aggressive interactions. Goats were classified as high (SI: 1–0.67), medium (SI: 0.66–0.34) and low-ranking (SI: 0.33–0.0). Ovulation was synchronized using two injections of prostaglandin 11 days apart, and the plasma progesterone levels determined daily for a period of 20 days. The area under the plasma progesterone curve during the entire study was greater in the high than in the medium and low-ranking does (96.2 ± 5.8, 79.5 ± 5.3 and 81.3 ± 5.3 ng/ml, respectively, P < 0.05). During days 11–17 following prostaglandin synchronization, the plasma progesterone levels were higher in the high-ranking (P < 0.05), compared to the low-ranking does. Plasma progesterone levels were significantly correlated with SI at days 14 and 15 (r = 0.26; P < 0.05). Results suggest a relationship between social ranking of goats and the plasma progesterone production from the corpus luteum and other possible sources.     

Photobiomodulation of human adipose-derived stem cells using 810 nm and 980 nm lasers operates via different mechanisms of action

Photobiomodulation (PBM) using red or near-infrared (NIR) light has been used to stimulate the proliferation and differentiation of adipose-derived stem cells. The use of NIR wavelengths such as 810 nm is reasonably well accepted to stimulate mitochondrial activity and ATP production via absorption of photons by cytochrome c oxidase. However, the mechanism of action of 980 nm is less well understood. Here we study the effects of both wavelengths (810 nm and 980 nm) on adipose-derived stem cells in vitro. Both wavelengths showed a biphasic dose response, but 810 nm had a peak dose response at 3 J/cm² for stimulation of proliferation at 24 h, while the peak dose for 980 nm was 10–100 times lower at 0.03 or 0.3 J/cm². Moreover, 980 nm (but not 810 nm) increased cytosolic calcium while decreasing mitochondrial calcium. The effects of 980 nm could be blocked by calcium channel blockers (capsazepine for TRPV1 and SKF96365 for TRPC channels), which had no effect on 810 nm. To test the hypothesis that the chromophore for 980 nm was intracellular water, which could possibly form a microscopic temperature gradient upon laser irradiation, we added cold medium (4 °C) during the light exposure, or pre-incubated the cells at 42 °C, both of which abrogated the effect of 980 nm but not 810 nm. We conclude that 980 nm affects temperature-gated calcium ion channels, while 810 nm largely affects mitochondrial cytochrome c oxidase.     

Role of ecosystem components in Cd removal process of aquatic ecosystem

An experiment was conducted using 15 glass aquariums to ascertain the pathways of removal of cadmium through numerical and compositional manipulation of ecosystem components and their role in Cd removal in different aquatic ecosystems. Each aquarium was provided with surface sediment @ 2 kg, filled with 15 L tap water and randomly distributed into five treatments having three replicates in each. Cadmium chloride (CdCl₂) of analytical grade was added @ 2 mg/L to the water of each aquarium and mixed gently. Except for the first one, the other four systems received unio (Lamellidens marginalis, 55 ± 2.5 g) @ 6 pieces/aquarium. Tilapia (Oreochromis mossambicus, 35 ± 3 g) was introduced @ 6 fish/aquarium in the third and fifth systems, whereas pistia (Pistia stratiotes) was introduced @ 50 g/aquarium in the fourth and fifth systems for a 28-day observation period. The samples of water, sediment, unio, fish and pistia were collected from different systems at 7-day intervals and analyzed. Results revealed that mean substantial reduction of Cd in water varied between 1.820 and 1.994 mg/L in different simulated ecosystems. Ecosystem efficiency of Cd removal varied in the different ecosystems and showed highest (11%) value in the ecosystem carrying five components, which suggested a cumulative effect of increasing number of components employed in different simulated aquatic ecosystems significantly facilitated the reduction of the level of Cd concentration in water column. Pistia exerted (12.88–547.5 times) higher rate of Cd accumulation over the other components employed in five simulated ecosystems of various component structures. Therefore, in the present study, it may be concluded that ecosystems carrying five components exhibited the best performance for optimum minimization of Cd removal from water column. It can also be concluded that ecosystem components showed a variable performance and pistia was the efficient component from the perspective of Cd removal.