RESUMO
Microalgae-based wastewater treatment (and biomass production) is an environmentally benign and energetically efficient technique as compared to traditional practices. The present study is focused on optimization of the major treatment variables such as temperature, light-dark cycle (LD), and nitrogen (N)-to-phosphate (P) ratio (N/P) for the elimination of N and P from tertiary municipal wastewater utilizing Chlorella kessleri microalgae species. In this regard, a hybrid support vector regression (SVR) technique integrated with the crow search algorithm has been applied as a novel modeling/optimization tool. The SVR models were formulated using the experimental data, which were furnished according to the response surface methodology with Box-Behnken Design. Various statistical indicators, including mean absolute percentage error, Taylor diagram, and fractional bias, confirmed the superior performance of SVR models as compared to the response surface methodology (RSM) and generalized linear model (GLM). Finally, the best SVR model was hybridized with the crow search algorithm for single/multi-objective optimizations to acquire the global optimal treatment conditions for maximum N and P removal efficiencies. The best-operating conditions were found to be 29.3°C, 24/0 h/h of LD, and 6:1 of N/P, with N and P elimination efficiencies of 99.97 and 93.48%, respectively. The optimized values were further confirmed by new experimental data.
Assuntos
Chlorella , Corvos , Microalgas , Purificação da Água , Algoritmos , Animais , Biomassa , Nitrogênio , Águas ResiduáriasRESUMO
Rapid, sensitive, on-site detection of bacteria without a need for sophisticated equipment or skilled personnel is extremely important in clinical settings and rapid response scenarios, as well as in resource-limited settings. Here, we report a novel approach for selective and ultra-sensitive multiplexed detection of Escherichia coli (non-pathogenic or pathogenic) using a lab-on-paper test strip (bioactive paper) based on intracellular enzyme (ß-galactosidase (B-GAL) or ß-glucuronidase (GUS)) activity. The test strip is composed of a paper support (0.5 × 8 cm), onto which either 5-bromo-4-chloro-3-indolyl-ß-D: -glucuronide sodium salt (XG), chlorophenol red ß-galactopyranoside (CPRG) or both and FeCl(3) were entrapped using sol-gel-derived silica inks in different zones via an ink-jet printing technique. The sample was lysed and assayed via lateral flow through the FeCl(3) zone to the substrate area to initiate rapid enzyme hydrolysis of the substrate, causing a change from colorless-to-blue (XG hydrolyzed by GUS, indication of nonpathogenic E. coli) and/or yellow to red-magenta (CPRG hydrolyzed by B-GAL, indication of total coliforms). Using immunomagnetic nanoparticles for selective preconcentration, the limit of detection was ~5 colony-forming units (cfu) per milliliter for E. coli O157:H7 and ~20 cfu/mL for E. coli BL21, within 30 min without cell culturing. Thus, these paper test strips could be suitable for detection of viable total coliforms and pathogens in bathing water samples. Moreover, inclusion of a culturing step allows detection of less than 1 cfu in 100 mL within 8 h, making the paper tests strips relevant for detection of multiple pathogens and total coliform bacteria in beverage and food samples.
Assuntos
Escherichia coli/isolamento & purificação , Papel , Colorimetria , Microbiologia de AlimentosRESUMO
We demonstrate a novel approach for rapid, selective, and sensitive detection of heavy metals using a solid-phase bioactive lab-on-paper sensor that is inkjet printed with sol-gel entrapped reagents to allow colorimetric visualization of the enzymatic activity of ß-galactosidase (B-GAL). The bioactive paper assay is able to detect a range of heavy metals, either alone or as mixtures, in as little as 10 min, with detection limits as follows: Hg(II) = 0.001 ppm; Ag(I) = 0.002 ppm, Cu(II) = 0.020 ppm; Cd(II) = 0.020 ppm; Pb(II) = 0.140 ppm; Cr(VI) = 0.150 ppm; Ni(II) = 0.230 ppm. The paper-based assay was immune to interferences from nontoxic metal ions such as Na(+) or K(+), could be used to detect heavy metals that were spiked into tap water or lake water, and provided quantitative data that was in agreement with values obtained by atomic absorption. With the incorporation of standard chromogenic metal sensing reagents into a multiplexed bioactive paper sensor, it was possible to identify specific metals in mixtures, albeit with much lower detection limits than were obtained with the enzymatic assay. The paper-based sensor should be valuable for rapid, on-site screening of trace levels of heavy metals in resource limited areas and developing countries.
Assuntos
Colorimetria/instrumentação , Metais Pesados/análise , Papel , beta-Galactosidase/metabolismo , Colorimetria/métodos , Ácido Edético/química , Escherichia coli/enzimologia , Metais Pesados/farmacologia , beta-Galactosidase/antagonistas & inibidores , beta-Galactosidase/químicaRESUMO
Several publications by authors in the field of petrochemical engineering have examined the use of chemically enhanced oil recovery (CEOR) technology, with a specific interest in polymer flooding. Most observations thus far in this field have been based on the application of certain chemicals and/or physical properties within this technique regarding the production of 50-60% trapped (residual) oil in a reservoir. However, there is limited information within the literature about the combined effects of this process on whole properties (physical and chemical). Accordingly, in this work, we present a clear distinction between the use of xanthan gum (XG) and hydrolyzed polyacrylamide (HPAM) as a polymer flood, serving as a background for future studies. XG and HPAM have been chosen for this study because of their wide acceptance in relation to EOR processes. To this degree, the combined effect of a polymer's rheological properties, retention, inaccessible pore volume (PV), permeability reduction, polymer mobility, the effects of salinity and temperature, and costs are all investigated in this study. Further, the generic screening and design criteria for a polymer flood with emphasis on XG and HPAM are explained. Finally, a comparative study on the conditions for laboratory (experimental), pilot-scale, and field-scale application is presented.
RESUMO
This study investigates the use of microalgae as a biosorbent to eliminate heavy metals ions from wastewater. The Chlorella kessleri microalgae species was employed to biosorb heavy metals from synthetic wastewater specimens. FTIR, and SEM/XRD analyses were utilized to characterize the microalgal biomass (the adsorbent). The experiments were conducted with several process parameters, including initial solution pH, temperature, and microalgae biomass dose. In order to secure the best experimental conditions, the optimum parameters were estimated using an integrated response surface methodology (RSM), desirability function (DF), and crow search algorithm (CSA) modeling approach. A maximum lead(II) removal efficiency of 99.54% was identified by the RSM-DF platform with the following optimal set of parameters: pH of 6.34, temperature of 27.71 °C, and biomass dosage of 1.5 g L-1. The hybrid RSM-CSA approach provided a globally optimal solution that was similar to the results obtained by the RSM-DF approach. The consistency of the model-predicted optimum conditions was confirmed by conducting experiments under those conditions. It was found that the experimental removal efficiency (97.1%) under optimum conditions was very close (less than a 5% error) to the model-predicted value. The lead(II) biosorption process was better demonstrated by the pseudo-second order kinetic model. Finally, simultaneous removal of metals from wastewater samples containing a mixture of multiple heavy metals was investigated. The removal efficiency of each heavy metal was found to be in the following order: Pb(II) > Co(II) > Cu(II) > Cd(II) > Cr(II).
Assuntos
Algoritmos , Biomassa , Chlorella/química , Metais Pesados/química , Microalgas/química , Modelos Químicos , Águas Residuárias/química , Purificação da ÁguaRESUMO
A reagentless bioactive paper-based solid-phase biosensor was developed for detection of acetylcholinesterase (AChE) inhibitors, including organophosphate pesticides. The assay strip is composed of a paper support (1 x 10 cm), onto which AChE and a chromogenic substrate, indophenyl acetate (IPA), were entrapped using biocompatible sol-gel derived silica inks in two different zones (e.g., sensing and substrate zones). The assay protocol involves first introducing the sample to the sensing zone via lateral flow of a pesticide-containing solution. Following an incubation period, the opposite end of the paper support is placed into distilled deionized water (ddH(2)O) to allow lateral flow in the opposite direction to move paper-bound IPA to the sensing area to initiate enzyme catalyzed hydrolysis of the substrate, causing a yellow-to-blue color change. The modified sensor is able to detect pesticides without the use of any external reagents with excellent detection limits (bendiocarb approximately 1 nM; carbaryl approximately 10 nM; paraoxon approximately 1 nM; malathion approximately 10 nM) and rapid response times (approximately 5 min). The sensor strip showed negligible matrix effects in detection of pesticides in spiked milk and apple juice samples. Bioactive paper-based assays on pesticide residues collected from food samples showed good agreement with a conventional mass spectrometric assay method. The bioactive paper assay should, therefore, be suitable for rapid screening of trace levels of organophosphate and carbamate pesticides in environmental and food samples.
Assuntos
Bebidas/análise , Técnicas Biossensoriais/métodos , Cromatografia em Papel/métodos , Contaminação de Alimentos/análise , Praguicidas/análise , Acetilcolinesterase/química , Acetilcolinesterase/metabolismo , Carbamatos/análise , Inibidores da Colinesterase/análise , Compostos Organofosforados/análiseRESUMO
There is an increasing interest in new strategies to rapidly detect analytes of clinical and environmental interest without the need for sophisticated instrumentation. As an example, the detection of acetylcholinesterase (AChE) inhibitors such as neurotoxins and organophosphates has implications for neuroscience, drug assessment, pharmaceutical development, and environmental monitoring. Functionalization of surfaces with multiple reagents, including enzymes and chromogenic reagents, is a critical component for the effective development of "dipstick" or lateral flow biosensors. Herein, we describe a novel paper-based solid-phase biosensor that utilizes piezoelectric inkjet printing of biocompatible, enzyme-doped, sol-gel-based inks to create colorimetric sensor strips. For this purpose, polyvinylamine (PVAm, which captures anionic agents) was first printed and then AChE was overprinted by sandwiching the enzyme within two layers of biocompatible sol-gel-derived silica on paper. AChE inhibitors, including paraoxon and aflatoxin B1, were detected successfully using this sensor by measuring the residual activity of AChE on paper, using Ellman's colorimetric assay, with capture of the 5-thio-2-nitrobenzoate (TNB(-)) product on the PVAm layer. The assay provided good detection limits (paraoxon, approximately 100 nM; aflatoxin B1, approximately 30 nM) and rapid response times (<5 min). Detection could be achieved either by eye or using a digital camera and image analysis software, avoiding the need for expensive and sophisticated instrumentation. We demonstrate that the bioactive paper strip can be used either as a dipstick or a lateral flow-based biosensor. The use of sol-gel-based entrapment produced a sensor that retained enzyme activity and gave reproducible results after storage at 4 degrees C for at least 60 days, making the system suitable for storage and use in the field.
Assuntos
Técnicas Biossensoriais/métodos , Neurotoxinas/análise , Inibidores da Colinesterase/química , Organofosfatos/química , Transição de Fase , Polivinil/química , Fitas Reagentes/química , Fitas Reagentes/metabolismoRESUMO
Monitoring extracellular metabolites of bacteria is very useful for not only metabolomics research but also for assessment of the effects of various chemicals, including antimicrobial agents and drugs. Herein, we describe the automated headspace solid-phase microextraction (HS-SPME) method coupled with gas chromatography-mass spectrometry (GC-MS) for the qualitative as well as semi-quantitative determination of metabolic responses of Escherichia coli to an antimicrobial agent, cinnamaldehyde. The minimum inhibitory concentration of cinnamaldehyde was calculated to be 2 g L(-1). We found that cinnamaldehyde was an important factor influencing the metabolic profile and growth process. A higher number of metabolites were observed during the mid-logarithmic growth phase. The metabolite variations (types and concentrations) induced by cinnamaldehyde were dependent on both cell density and the dose of cinnamaldehyde. Simultaneously, 25 different metabolites were separated and detected (e.g., indole, alkane, alcohol, organic acids, esters, etc.) in headspace of complex biological samples due to intermittent addition of high dose of cinnamaldehyde. The study was done using an automated system, thereby minimizing manual workup and indicating the potential of the method for high-throughput analysis. These findings enhanced the understanding of the metabolic responses of E. coli to cinnamaldehyde shock effect and demonstrated the effectiveness of the SPME-GC-MS based metabolomics approach to study such a complex biological system.
Assuntos
Escherichia coli/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Metaboloma , Compostos Orgânicos Voláteis/análise , Acroleína/análogos & derivados , Acroleína/farmacologia , Automação , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microextração em Fase Sólida , Compostos Orgânicos Voláteis/isolamento & purificação , Compostos Orgânicos Voláteis/metabolismoRESUMO
This paper reports the development of a method to control the flow rate of fluids within paper-based microfluidic analytical devices. We demonstrate that by simply sandwiching paper channels between two flexible films, it is possible to accelerate the flow of water through paper by over 10-fold. The dynamics of this process are such that the height of the liquid is dependent on time to the power of 1/3. This dependence was validated using three different flexible films (with markedly different contact angles) and three different fluids (water and two silicon oils with different viscosities). These covered channels provide a low-cost method for controlling the flow rate of fluid in paper channels, and can be added following printing of reagents to control fluid flow in selected fluidic channels. Using this method, we redesigned a previously published bidirectional lateral flow pesticide sensor to allow more rapid detection of pesticides while eliminating the need to run the assay in two stages. The sensor is fabricated with sol-gel entrapped reagents (indoxyl acetate in a substrate zone and acetylcholinesterase, AChE, in a sensing zone) present in an uncovered "slow" flow channel, with a second, covered "fast" channel used to transport pesticide samples to the sensing region through a simple paper-flap valve. In this manner, pesticides reach the sensing region first to allow preincubation, followed by delivery of the substrate to generate a colorimetric signal. This format results in a uni-directional device that detects the presence of pesticides two times faster than the original bidirectional sensors.
Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Organofosfatos/análise , Papel , Praguicidas/análise , Acetilcolinesterase/metabolismo , Animais , Electrophorus , Proteínas de Peixes/metabolismo , Indóis/química , Modelos Teóricos , Fatores de TempoRESUMO
This protocol represents a novel enzyme-luminescence method to detect dopamine sensitively and rapidly with high temporal resolution. In principle, dopamine is first oxidized with tyramine oxidase to produce H(2)O(2), and then the produced H(2)O(2) reacts with luminol to generate chemiluminescence in the presence of horseradish peroxidase (POD). We applied this method successfully to perform real-time monitoring of dopamine release from PC12 cells using a luminescence plate reader upon stimulation with several drugs (e.g., acetylcholine, bradykinin). The results indicated that the dopamine release from PC12 cells was modulated by these drugs in a way similar to that found by using several conventional analytical techniques, such as HPLC-electrochemical detector (ECD). Unlike other assays, this assay technique is simple, rapid, highly sensitive and thus useful for assessment of effects of drugs on the nervous system. The dopamine release assay takes only < or =1 h once reagent setup and culture plates' preparation are finished.