Screening and identification of newly isolated Pseudomonas sp. for biodegrading the textile azo dye C.I. Procion Red H-3B.

AIM: To test the potential of a newly isolated strain of Pseudomonas sp., and its optimization for carrying out bioremediation of textile azo dye Procion Red H-3B. METHOD: The isolation of the bacterial strain was done from a textile waste dumping site, followed by screening techniques to study the decolourization of an azo dye. The isolated pure culture was selected by its ability to form clear zones. The biochemical tests gave partial confirmation of the isolates, and the phylogenic analysis made the complete confirmation by 16S rRNA sequencing. RESULT: The identified strain belongs to the genus Pseudomonas. The phylogenic analysis confirmed that the strain belongs to Pseudomonas stutzeri. The culture exhibited maximum decolourization at pH between 6 and 8, the optimum at pH 7·5 and 37°C temperature. A maximum of 96% discolouration was observed at 50 mg l-1 of initial dye concentration after 24 h of incubation period. At a dye concentration equally or greater than 600 mg l-1 , the colour removal was drastically decreased to 30%. The use of fructose at 1% (w/v) and peptone 0·5% (w/v) concentration for 24 h of incubation, as carbon and nitrogen source, showed luxuriant decolourization. The results showed that the Pseudomonas sp. holds immense potential in treating textile effluents containing the dye Procion red H-3B. CONCLUSION: Pseudomonas is a known organism in bioremediation of various textile dyes but not much has being reported about the role of P. stutzeri in the bioremediation of azo dyes. This study revealed the immense potential of this strain in degrading the azo dyes. SIGNIFICANCE AND IMPACT OF THE STUDY: The strain shows prospective for industrial application in the field of textile wastewater treatment. Bioremediation is comparatively cheaper and more effective treatment, thus holds promising future for a cleaner environment.

Electromagnetic dispersive solid-phase extraction based on polyaniline-coated magnetite/silica materials for the determination of Sudan red I in drinks.

By replacing the permanent magnet with an electromagnet, traditional magnetic solid-phase extraction was developed into electromagnetic dispersive solid-phase extraction. A simple operation of power on and off can realize the separation of adsorbents from solutions easily. The improvement makes it possible for the automation of the determination of Sudan Red I by high-performance liquid chromatography. After series of optimization, a trace amount of Sudan red I was well-determined, and excellent linearity was achieved in the range of 0.005 to 1 mg/L with the correlation coefficient (R2 ) = 0.997. The limit of detection with a signal-to-noise ratio of 3 was found to be 0.001 mg/L. The spiked recoveries of Sudan red I for the samples ranged from 93.6 to 104.9%. Moreover, the adsorbent could be recycled at least ten times. The results show that the electromagnetic dispersive solid-phase extraction combined with high-performance liquid chromatography is a rapid, eco-friendly, effective, and sensitive determination method with fascinating automation potential and high practical applicability.

A tunable assay for modulators of genome-destabilizing DNA structures.

Regions of genomic instability are not random and often co-localize with DNA sequences that can adopt alternative DNA structures (i.e. non-B DNA, such as H-DNA). Non-B DNA-forming sequences are highly enriched at translocation breakpoints in human cancer genomes, representing an endogenous source of genetic instability. However, a further understanding of the mechanisms involved in non-B DNA-induced genetic instability is needed. Small molecules that can modulate the formation/stability of non-B DNA structures, and therefore the subsequent mutagenic outcome, represent valuable tools to study DNA structure-induced genetic instability. To this end, we have developed a tunable Förster resonance energy transfer (FRET)-based assay to detect triplex/H-DNA-destabilizing and -stabilizing ligands. The assay was designed by incorporating a fluorophore-quencher pair in a naturally-occurring H-DNA-forming sequence from a chromosomal breakpoint hotspot in the human c-MYC oncogene. By tuning triplex stability via buffer composition, the assay functions as a dual-reporter that can identify stabilizers and destabilizers, simultaneously. The assay principle was demonstrated using known triplex stabilizers, BePI and coralyne, and a complementary oligonucleotide to mimic a destabilizer, MCRa2. The potential of the assay was validated in a 384-well plate with 320 custom-assembled compounds. The discovery of novel triplex stabilizers/destabilizers may allow the regulation of genetic instability in human genomes.

Development of standardized method for the quantification of azo dyes by UV-Vis in binary mixtures.

The azo dyes, Yellow 5 (Y5), Red 2 (R2) and Blue 1 (B1), quantified in solutions and in mixtures of binary dyes, were studied by means of UV-Vis spectroscopy. In this work was used a CIE algorithm developed in Visual Basic for Applications (VBA). The CIE algorithm is based on the tristimulus chromaticity diagram, as an alternative to the shielding effect that arises in dye mixtures, and it can also be applied to complex quantification methods such as HPLC (High Performance Liquid Chromatography). The results obtained through of the algorithm, showed a higher accuracy from 97 to 99% in relation with similar UV-Vis quantification methods. In contrast, linear methods only managed to reach an accuracy from 78 to 98%. Additionally, the algorithm yielded significant similar values to the UHPLC reference method. The results showed that the method CIE algorithm was accessible and reliable to quantify binary mixtures of the dyes used which suggests the possibility to apply this method on other dyes, within the limits of quantification obtained in this study (0.076-24.56 mg/L) and the pH values from 2 to 10.

Flower-like layered double hydroxide-modified stainless-steel fibers for online in-tube solid-phase microextraction of Sudan dyes.

Layered double hydroxides are a family of inorganic crystals that have gained a lot of attention due to its special structure and properties such as high porosity, large specific area, and excellent anion exchange ability. In this work, flower-like NiAl-layered double hydroxides with high specific area were in situ immobilized onto the stainless steel fibers by bioinspired polydopamine modification method and packed into poly (ether ether) ketone tube for online solid-phase microextraction with high performance liquid chromatography analysis. Thanks to the high specific surface area and excellent extraction ability of the NiAl-layered double hydroxides, the fibers showed excellent extraction performance to three Sudan dyes with enrichment factors between 260 to 650 folds. After optimization of the reaction and extraction conditions, an online solid-phase microextraction method was developed for determination of Sudan dyes in water samples and chili samples. The method has limits of detection of 0.01 to 0.02 ng/mL, good linearity and good reproducibility (≤1.45%).

Integrated metagenomic and metaproteomic analyses reveal potential degradation mechanism of azo dye-Direct Black G by thermophilic microflora.

Direct Black G (DBG) is a typical toxic azo dye with extensive applications but it poses a serious threat to the aquatic ecosystem and humans. It is necessary to efficiently and safely remove DBG from environments by the application of various treatment technologies. A thermophilic microflora previously isolated from the soil can effectively metabolize DBG. However, the molecular basis of DBG degradation by this thermophilic microflora remains unknown. In this study, metagenomic sequencing technology and qRT-PCR have been used to elucidate the functional potential of genes and their modes of action on DBG. A quantitative metaproteomic method was further utilized to identify the relative functional proteins involved. Subsequently, the possible co-metabolic molecular mechanisms of DBG degradation by candidate genes and functional proteins of the thermophilic microflora were illustrated. The combination of metagenomics and metaproteomics to investigate the degradation of DBG by a microflora was reported for the first time in recent literature; this can further provide a deep insight into the molecular degradation mechanism of dye pollutants by natural microflora.

Biochar-activated persulfate for organic contaminants removal: Efficiency, mechanisms and influencing factors.

Turning biomass into biochar as a multifunctional carbon-based material for water remediation has attracted much research attention. Sawdust and rice husk were selected as feedstock for biochar (BC) production, aiming to explore their performance as a catalyst to activate persulfate (PS) for degrading acid orange 7 (AO7). There was an excellent synergistic effect in the combined BC/PS system. Sawdust biochar (MX) showed a faster and more efficient performance for the AO7 degradation due to its abundant oxygen functional groups, compared to rice husk biochar (DK). In the BC/PS system, AO7 was well decolorized and mineralized. Based on the two-dimensional correlation analysis method, the azo conjugation structure and naphthalene ring of AO7 molecule changed first then benzene ring changed during the reaction. Moreover, AO7 decolorization efficiency increased with the increase of PS concentration and biochar dosage, and the deacrease of pH. Biochar deactivated after used twice. When the biochar reached its adsorption equilibrium of AO7, the AO7 could not be degraded in the BC/PS system. SO4- and OH participated in the reaction together and OH played the main role in activating PS to AO7 decolorization based on the radical scavengers experiment. All of results indicate using biochar to activate PS for degradation of AO7 contaminated water is a promising method.

Response of Trametes hirsuta to hexavalent chromium promotes laccase-mediated decolorization of reactive black 5.

The recalcitrant azo dyes combined with heavy metals constitute a major challenge for the bioremediation of industrial effluents. This study aimed to investigate the effect and mechanism of action of a white-rot fungus Trametes hirsuta TH315 on the simultaneous removal of hexavalent chromium [Cr(VI)] and azo dye (Reactive Black 5, RB5). Here, this study discovered that toxic Cr(VI) (1 mM) greatly promoted RB5 decolorization (from 57.15% to 83.65%) by white-rot fungus Trametes hirsuta with high Cr(VI)-reducing ability (>96%), resulting in the simultaneous removal of co-contaminants. On the basis of transcriptomic and biochemical analysis, our study revealed that the oxidative stress in co-contaminants mainly caused by Cr(VI), and a number of dehydrogenases and oxidases showed up-regulation in response to Cr(VI) stress. It was noteworthy that the oxidative stress caused by Cr(VI) in co-contaminants can both significantly induce glutathione S-transferase and laccase expression. Glutathione S-transferase potentially involved in antioxidation against Cr(VI) stress. Laccase was found to play a key role in RB5 decolorization by T. hirsuta. These results suggested that the simultaneous removal of co-contaminants by T. hirsuta could be achieved with Cr(VI) exposure. Overall, the elucidation of the molecular basis in details will help to advance the general knowledge about the fungus by facing harsh environments, and put forward a further possible application of fungi on environmental remediation.

Genome and transcriptome analysis of a newly isolated azo dye degrading thermophilic strain Anoxybacillus sp.

Understanding azo dye degrading enzymes and the encoding of their functional genes is crucial for the elucidation of their molecular mechanisms. In this study, a thermophilic strain capable of degrading azo dye was isolated from the soil near a textile dye manufacturing factory. Based on its morphological, physiological and biochemical properties, as well as 16S rRNA gene sequence analysis, the strain was identified as Anoxybacillus sp. PDR2. The decolorization ratios of 100-600 mg/L Direct Black G (DBG) by strain PDR2 reached 82.12-98.39% within 48 h of dyes. Genome analysis revealed that strain PDR2 contains a circular chromosome of 3791144 bp with a G + C content of 42.48%. The genetic basis of azo dye degradation by strain PDR2 and its capacity to adapt to harsh environments, were further elucidated through bioinformatics analysis. RNA-Seq and qRT-PCR technology confirmed that NAD(P)H-flavin reductase, 2Fe-2S ferredoxin and NAD(P)-dependent ethanol dehydrogenase genes expressed by strain PDR2, were the key genes involved in DBG degradation. The combination of genome and transcriptome analysis was utilized to explore the key genes of strain PDR2 involved in azo dye biodegradation, with these findings providing a valuable theoretical basis for the practical treatment of azo dye wastewater.

Mitigation of the inhibitory effects of co-existing substances on the Fenton process by UV light irradiation.

Co-existing substances (substances not targeted for degradation) can negatively affect wastewater treatment process performance. Here, we quantitatively evaluated the effects of propanal, a common co-existing substance, on the degradation of the azo-dye Orange II, a common pollutant, by the Fenton process to provide data for the development of measures to reduce the effects of co-existing substances on this wastewater treatment process. Inhibition rate (IR; ratio of the reaction rate constants obtained in the absence and presence of propanal) was calculated to examine the effects of propanal on the degradation of Orange II. The IRs for the Fenton process in the first phase and the second phase were 1.6 and 4.2, respectively. However, addition of ultraviolet irradiation to the Fenton process (i.e., the photo-Fenton process) resulted in a comparable IR for the first phase but a markedly lower IR for the second phase. We attributed this to the improvement of the photo-reduction reaction rate due to complexation of propanal with ferric ions, which compensated for the scavenger effects (the trapping of OH radicals) of propanal. Thus, ultraviolet irradiation reduced the inhibitory effects of propanal on the degradation of Orange II by the Fenton process.

Electrochemical detection and quantification of Reactive Red 195 dyes on graphene modified glassy carbon electrode.

Reactive Red 195 was detected from industrial waste samples electrochemically on graphene modified glassy carbon electrode (GCE), using both bare and surface changed GCE at different pH media from 1.0 to 13.0. The optimum pH was determined to be 4.0. RR 195 exhibited good linear responds at pH 4.0 on both electrode surfaces. Other parameters, such as accumulation potential, accumulation time, initial scan potential, pulse height, pulse width, and potential scan increment and scan rate are optimized and calibration plot was also derived on different concentrations of the dye. The stripping voltammetric behavior of dye exhibits very low limit of detection on graphene coated electrode (30 ppb). The adsorption of compound on GCE and graphene coated GCE are confirmed using atomic force microscopy studies.

Highly sensitive and rapid determination of sunset yellow in drinks using a low-cost carbon material-based electrochemical sensor.

Starting from simple graphite flakes, an electrochemical sensor for sunset yellow monitoring is developed by using a very simple and effective strategy. The direct electrochemical reduction of a suspension of exfoliated graphene oxide (GO) onto a glassy carbon electrode (GCE) surface leads to the electrodeposition of electrochemically reduced oxide at the surface, obtaining GCE/ERGO-modified electrodes. They are characterized by cyclic voltammetry (CV) measurements and field emission scanning electron spectroscopy (FE-SEM). The GCE/ERGO electrode has a high electrochemically active surface allowing efficient adsorption of SY. Using differential pulse voltammetry (DPV) technique with only 2 min accumulation, the GCE/ERGO sensor exhibits good performance to SY detection with a good linear calibration for concentration range varying 50-1000 nM (R2 = 0.996) and limit of detection (LOD) estimated to 19.2 nM (equivalent to 8.9 µg L-1). The developed sensor possesses a very high sensitivity of 9 µA/µM while fabricated with only one component. This electrochemical sensor also displays a good reliability with RSD value of 2.13% (n = 7) and excellent reusability (signal response change < 3.5% after 6 measuring/cleaning cycles). The GCE/ERGO demonstrates a successful practical application for determination of sunset yellow in commercial soft drinks. Graphical abstract.

Hydroxamic acid mediated heterogeneous Fenton-like catalysts for the efficient removal of Acid Red 88, textile wastewater and their phytotoxicity studies.

Heterogeneous Fenton-like catalyst and its industrial application are increasingly given importance for its non-selective mineralization of organic pollutants in broad pH range. Current study, utilized an aromatic hydroxamic acid derivative 2-hydroxypyridine-N-oxide (HpO), for the construction of iron-Hpo ligand catalyst supported on granular activated carbon (GAC). 8-Hydroxyquinoline and citric acid as non-hydroxamic aromatic and aliphatic Fenton-like catalysts were used for comparative evaluation of the efficiency with targeted catalyst (iron-HpO-GAC). This novel catalyst iron-HpO-GAC exhibits excellent efficiency in Acid Red 88 dye removal in the presence of hydrogen peroxide as oxidant at acidic, basic as well as at neutral conditions. Operational conditions for the catalytic oxidation including temperature, dye concentration, pH and catalyst dosage were systematically investigated and analyzed through kinetic studies. Thermodynamic analysis of the catalytic dye removal revealed that the system could oxidize pollutants faster with less activation energy requirement. Higher level of recyclability and stability of the catalyst with less iron leaching was achieved. Finally, the real time application of the catalyst was investigated through successful repeated treatment for actual industrial wastewater. The phytotoxicity assay (with respect to plant Phaseolus mungo) revealed that the degradation of Acid Red 88 and dye wastewater produced nontoxic metabolites which increases its potential application. This study emphasizes the viability of hydroxamate mediated efficient Fenton-like oxidation as a novel approach in designing economically viable pollutant removal technology.

Studies on the reaction mechanism of Cu/SiC catalytic oxidation for degradation of methyl orange in presence of microwave.

The removal of methyl orange (MO) in a copper-loaded silicon carbide (Cu/SiC) system under microwave (MW) irradiation was studied. Cu/SiC was synthesized by employing an impregnation method and the effects of parameters such as reaction time, catalyst dosage, hydrogen peroxide (H2O2) dosage, microwave power and pH on the rate of degradation of MO were also studied. The obtained results showed that almost complete degradation was obtained in the presence of Cu/SiC catalyst within 8 min of irradiation when 100 mL of MO solution (20 mg/L), 3 ml/L of H2O2, 2 g/L of catalyst dose, 600 W of MW power, and pH 7 were applied. The Cu-bearing catalyst with H2O2 formed a Fenton-like system and the rate of generation of hydroxyl radicals (·OH) was also accelerated by subjecting to MW. From the kinetic analysis, it is revealed that the degradation of MO using the MW-Cu/SiC-H2O2 system follows the pseudo-first-order.

Decolorization effect and related mechanism of atmospheric pressure plasma jet on Eriochrome Black T.

In this study, Eriochrome Black T (EBT) in water was decolorized by means of argon atmospheric pressure plasma jet (APPJ), which showed great decolorization performance. The results showed that the relatively high decolorization rate (approximately 80%) was obtained after plasma treatment for 6 min. Changes to some reactive oxygen and nitrogen species (RONS) in the liquid phase were detected. The contents of peroxide, HO·, O2 -·, and NO· in the plasma-treated EBT solution were much less than those in the activated water. The roles of H2O2 and HO· in the decolorization of EBT solution were explored by evaluating the effects of their scavengers, and by exploring the direct effect of H2O2. The results indicated that reactive oxygen species (ROS), especially HO· and O2 -·, played significant roles in the decolorization of the EBT solution. Analysis of degradation by-products indicated that plasma discharge could destroy the azo bond first and gradually break the aromatic rings of EBT molecules into small molecular compounds.

Pre-magnetization for enhancing the iron-catalyzed activation of peroxymonosulfate via accelerating the corrosion of Fe0.

Our findings proved that micron-scale zero-valent iron (mZVI) particles with pre-magnetization combined with peroxymonosulfate (PMS) can markedly enhance the removal of acid orange 7 (AO7). Investigation into the mechanism showed that PMS accelerated the corrosion of ZVI to release Fe2+ under acidic conditions, and the in-situ generated Fe2+ further activated PMS to produce SO4•- and •OH, resulting in AO7 removal. Further, the Lorentz force strengthened the convection in the solution and the field gradient force tended to move Fe2+ from a higher to a lower field gradient at the pre-magnetized ZVI (Pre-ZVI) particle surfaces, thus indicating that pre-magnetization promoted the corrosion of ZVI to release Fe2+, which resulted in the enhancement of PMS activation. Nano-scale ZVI (nZVI) was more effective than mZVI in activating PMS to degrade AO7, but the pre-magnetization effect on mZVI was better than on nZVI. AO7 removal increased with higher ZVI and PMS dosage, lower AO7 concentration, and acidic conditions (pH = 2, 3). This study helps to understand the reactive radicals-based oxidation process with application of pre-magnetized ZVI in activating PMS.

Co3O4/TiO2 hetero-structure for methyl orange dye degradation.

Advanced oxidation processes based on sulphate radical generated by peroxymonosulphate (PMS) activation is a promising area for environmental remediation. One of the biggest drawbacks of heterogeneous PMS activation is catalyst instability and metal ion leaching. In this study, a simple organic binder mediated route was explored to substitute Ti4+ ions into the Co3O4 host lattice structure to create a Co-O-Ti bond to minimise cobalt leaching during methyl orange degradation. The catalyst was characterised by X-ray diffraction, and scanning and transmission electron microscopy. The as-prepared catalysts with Co3O4:TiO2 ratio of 70:30 exhibited minimal leaching (0.9 mg/L) compared to other ratios studied. However, the pristine Co3O4 exhibited highest catalytic activity (rate constant = 0.41 min-1) and leaching (26.7 mg/L) compared to composite material (70:30 Co3O4:TiO2). Interestingly, the morphology of the composite and leaching of Co2+ ions were found to be temperature dependent, as an optimum temperature ensured strong Co-O-Ti bond for prevention of Co2+ leaching. The classical quenching test was utilised to determine the presence and role of radical species on methyl orange degradation. The fabricated catalyst also exhibited good catalytic activity in degrading mixed dyes and good recyclability, making it a potential candidate for commercial application.

Optimization and mechanism of Acid Orange 7 removal by powdered activated carbon coupled with persulfate by response surface method.

In this study, powder activated carbon (PAC) utilized to activate peroxydisulfate (PDS) was investigated for decolorization of Acid Orange 7 (AO7). The results indicated a remarkable synergistic effect in the PAC/PDS system. The effect of PAC, PDS dosages and initial pH on AO7 decolorization were studied and the processes followed first-order kinetics. Response surface method with central composite design (CCD) model was utilized to optimize these three factors and analyze the combined interaction. The optimum condition for the decolorization rate of AO7 was analyzed as the following: PAC (0.19 g/L), PDS (1.64 g/L), and initial pH (4.14). Cl- and SO4 2- showed a promoting effect on AO7 decolorization while HCO3 - had a slightly inhibiting effect. Quenching experiments confirmed that both sulfate and hydroxyl radicals were the oxidizing species, and the oxidation reaction occurred on the surface of PAC. The results of UV-vis spectrum with 100% decolorization rate and the 50% total organic carbon reduction indicated highly efficient decolorization and mineralization of AO7 in the PAC/PDS system. Finally, the recovery performance of PAC was studied and the result indicated PAC had poor reuse in reactivity.

Process optimization for effective bio-decolourization of reactive orange 16 using chemometric methods.

Azo group containing reactive dyes are most commonly used in textile and tannery industries due to its bright appearance and stable color. This study aims to investigate the decolourization of reactive orange 16 (RO16) dye by Pseudomonas aeruginosa 23N1 along with removal of chromate (Cr(VI)) and evaluation of optimal process condition. The regular two-level factorial design is used to screen out operational parameters and selects their levels for further optimization process through central composite design (CCD) based response surface methodology (RSM). The result revealed that glucose and peptone have a negative effect on the performance of dye decolourization. Bacteria exhibit high decolourization potential in yeast extract supplemented culture medium with no addition of external carbon sources. The percentages of decolourization obtained in model validated experiments are obtained as 95.0 ± 0.4% and 95.1 ± 0.5% for initial dye 50 mg/L and 150 mg/L, respectively, which exhibit satisfactory correlation with model predicted response. The simultaneous dye and Cr(VI) removal has been explored in this study. The decolourization of dye is only affected due to presence of high Cr(VI) concentration (>120 mg/L). Bacteria have shown satisfactorily decolourization for RO16 contaminated industrial wastewater. The strain 23N1 could be a good biological agent for decolourization of RO16 dye.

Development of aptamer fluorescent switch assay for aflatoxin B1 by using fluorescein-labeled aptamer and black hole quencher 1-labeled complementary DNA.

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins and draws great concern in health and food safety. A DNA aptamer against AFB1 having a stem-loop structure shows high binding affinity to AFB1 and promise in assay development for AFB1 detection. Based on the structure-switching property of the aptamer, we report an aptamer fluorescence assay for AFB1 detection. Aptamer with fluorescein (FAM) label at 5' end was used as affinity ligand, while its short complementary DNA (cDNA) with BHQ1 (black hole quencher 1) label at 3' end was used as a quencher. In the absence of AFB1, FAM-labeled aptamer hybridized with BHQ1-labeled cDNA, forming a duplex of cDNA and aptamer, resulting in fluorescence quenching of FAM. When AFB1 bound with aptamer, the BHQ1-labeled cDNA was displaced from aptamer, causing fluorescence restoration of FAM. We tested a series of FAM-labeled aptamers and BHQ1-labeled cDNAs with different lengths. The lengths of the aptamer stem and the cDNA, Mg2+ in binding buffer, and temperature had significant influence on the performance of the assay. Under optimized conditions, we achieved sensitive detection of AFB1 by using a 29-mer FAM-labeled aptamer and a 14-mer BHQ1-labeled cDNA, and the detection limit of AFB1 reached 0.2 nM. The maximum fluorescence recovery rate of FAM-labeled aptamer caused by AFB1 was about 69-fold. This method enabled the detection of AFB1 in complex sample matrix, e.g., diluted wine samples and maize flour samples. This aptamer-based fluorescent assay for AFB1 determination shows potential for broad applications. Graphical abstract ᅟ.