The bioenergetics mechanisms and applications of sulfate-reducing bacteria in remediation of pollutants in drainage: A review.

Sulfate-reducing bacteria (SRB), a group of anaerobic prokaryotes, can use sulfur species as a terminal electron acceptor for the oxidation of organic compounds. They not only have significant ecological functions, but also play an important role in bioremediation of contaminated sites. Although numerous studies on metabolism and applications of SRB have been conducted, they still remain incompletely understood and even controversial. Fully understanding the metabolism of SRB paves the way for allowing the microorganisms to provide more beneficial services in bioremediation. Here we review progress in bioenergetics mechanisms and application of SRB including: (1) electron acceptors and donors for SRB; (2) pathway for sulfate reduction; (3) electron transfer in sulfate reduction; (4) application of SRB for economical and concomitant treatment of heavy metal, organic contaminants and sulfates. Moreover, current knowledge gaps and further research needs are identified.

A fluorescent DNA based probe for Hg(II) based on thymine-Hg(II)-thymine interaction and enrichment via magnetized graphene oxide.

The authors describe a fluorometric assay for the determination of Hg(II). A naphthalimide derivative is used as a label for a thymine (T) rich ssDNA, and graphene oxide magnetized with Fe3O4 nanoparticles acts as a quencher and preconcentrators. In the absence of Hg(II), the labeled ssDNA does not separate from the magnetized graphene oxide. As a result, fluorescence is fully quenched. In the presence of Hg(II), a T-Hg(II)-T link is formed dues to the highly affinity between T and Hg(II). Hence, fluorescence is restored. The assay has a linear response in the 1.0 to 10.0 nM Hg(II) concentration range, and a 0.65 nM detection limit. The method is selective and sensitive. It was applied to the analysis of spiked environmental water samples, and data agreed well with those obtained by atomic fluorescence spectrometry. Graphical abstract Strategy of a fluorescent probe for detecting Hg(II). The method has a 0.65 nM detection limit and is selective. MGO: magnetized graphene oxide, AHN: a fluorescent derivative of naphthalimide.

A Fluorescence Sensor for Lead (II) Ions Determination Based on Label-Free Gold Nanoparticles (GNPs)-DNAzyme Using Time-Gated Mode in Aqueous Solution.

This paper describes a label-free 17E DNAzyme-based time-gated fluorescence sensor for Pb2+ detection by unmodified gold nanoparticles (GNPs) and a terbium ternary complex. The fluorophore that used in this paper is a terbium ternary complex. Its signal can be measured in a time-gated manner which could eliminate most of the unspecific fluorescent background. It is well known that unfolded single-stranded DNA (ssDNA) could be adsorbed on GNPs while double-stranded DNA could not. The cleavage of the substrate by the 17E DNAzyme in the presence of Pb2+ causes the release of ssDNA from the 17E-17S duplex to be absorbed onto GNPs, preventing the aggregation of GNPs and then leading to a fluorescence decrease of terbium ternary complex. By means of this method, the authors have successfully detected Pb2+ over a range of 10 nM to 2500 nM with a detection limit of 1.7 nM. The sensor also exhibited good selectivity. The sensor provided a simple, cost-effective, rapid and sensitive measurement tool for Pb2+ detection.

An exploration of spatial human health risk assessment of soil toxic metals under different land uses using sequential indicator simulation.

A modified method was proposed which integrates the spatial patterns of toxic metals simulated by sequential indicator simulation, different exposure models and local current land uses extracted by remote-sensing software into a dose-response model for human health risk assessment of toxic metals. A total of 156 soil samples with a various land uses containing farm land (F1-F25), forest land (W1-W12) and residential land (U1-U15) were collected in a grid pattern throughout Xiandao District (XDD), Hunan Province, China. The total Cr and Pb in topsoil were analyzed. Compared with Hunan soil background values, the elevated concentrations of Cr were mainly located in the east of XDD, and the elevated concentrations of Pb were scattered in the areas around F1, F6, F8, F13, F14, U5, U14, W2 and W11. For non-carcinogenic effects, the hazard index (HI) of Cr and Pb overall the XDD did not exceed the accepted level to adults. While to children, Cr and Pb exhibited HI higher than the accepted level around some areas. The assessment results indicated Cr and Pb should be regarded as the priority pollutants of concern in XDD. The first priority areas of concern were identified in region A with a high probability (>0.95) of risk in excess of the accepted level for Cr and Pb. The areas with probability of risk between 0.85 and 0.95 in region A were identified to be the secondary priority areas for Cr and Pb. The modified method was proved useful due to its improvement on previous studies and calculating a more realistic human health risk, thus reducing the probability of excessive environmental management.

Biochar pyrolyzed from MgAl-layered double hydroxides pre-coated ramie biomass (Boehmeria nivea (L.) Gaud.): Characterization and application for crystal violet removal.

A novel biochar/MgAl-layered double hydroxides composite (CB-LDH) was prepared for the removal of crystal violet from aqueous solution by pyrolyzing MgAl-LDH pre-coated ramie stem (Boehmeria nivea (L.) Gaud.). Pyrolysis played dual role for both converting biomass into biochar and calcining MgAl-LDH during the pyrolysis process. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive X-ray analysis (EDS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared (FTIR) and zeta potential analysis were used to characterize the CB-LDH. The results of characterization suggested that the calcined LDH was successfully synthesized and coated on biochar. The resulted CB-LDH had higher total pore volume and more functional groups than the pristine biochar. Adsorption experimental data fitted well with the pseudo-second order kinetics model and the Freundlich isotherm model. The rate-controlled step was controlled by film-diffusion initially and then followed by intra-particle diffusion. Thermodynamic analysis showed that the adsorption of crystal violet was a spontaneous and endothermic process. The higher pH and temperature of the solution enhanced the adsorption performance. CB-LDH could also have excellent ability for the removal of crystal violet from the actual industrial wastewater and groundwater with high ionic strength. LDH adsorption, electrostatic attraction, pore-filling, π-π interaction and hydrogen bond might be the main mechanisms for crystal violet adsorption on CB-LDH. The results of this study indicated that CB-LDH is a sustainable and green adsorbent with high performance for crystal violet contaminated wastewater treatment and groundwater remediation.

Electrochemical sensor based on electrodeposited graphene-Au modified electrode and nanoAu carrier amplified signal strategy for attomolar mercury detection.

An electrochemical sensor was developed for attomolar Hg(2+) detection. Three single-stranded DNA probes were rationally designed for selective and sensitive detection of the target, which combined T-Hg(2+)-T coordination chemistry and the characteristic of convenient modification of electrochemical signal indicator. Graphene and nanoAu were successively electrodeposited on a glass carbon electrode surface to improve the electrode conductivity and functionalize with the 10-mer thymine-rich DNA probe (P1). NanoAu carriers functionalized with 29-mer guanine-rich DNA probe (P3) labeled methyl blue (MB-nanoAu-P 3s) were used to further strengthen signal response. In the presence of Hg(2+), a T-T mismatched dsDNA would occur between P1 and a 22-mer thymine-rich DNA probe (P2) on the electrode surface due to T-Hg(2+)-T coordination chemistry. Followed by adding the MB-nanoAu-P 3s for hybridization with P2, square wave voltammetry was executed. Under optimal conditions, Hg(2+) could be detected in the range from 1.0 aM to 100 nM with a detection limit of 0.001 aM. Selectivity measurements reveal that the sensor is specific for Hg(2+) even with interference by high concentrations of other metal ions. Three different environmental samples were analyzed by the sensor and the results were compared with that from an atomic fluorescence spectrometry. The developed sensor was demonstrated to achieve excellent detectability. It may be applied to development of ultrasensitive detection strategies.

Micellar-enhanced ultrafiltration for the solubilization of various phenolic compounds with different surfactants.

Micellar-enhanced ultrafiltration (MEUF) was applied to the separation of phenolic compounds p-nitrophenol (PNP), p-chlorophenol (PCP), p-cresol (PC) and phenol (P) from effluents using a hydrophilic polyethersulfone ultrafiltration membrane. Cationic cetylpyridinium chloride (CPC), nonionic TX-100 and anionic sodium dodecyl benzene sulfonate (SDBS) were chosen as the surfactants. Several important parameters, i.e. the separation efficiency, the distribution coefficient of phenolic compounds and the removal ratio of surfactants, were investigated. It was shown that the separation efficiency and the distribution coefficient of phenolic compounds ascended with the increasing surfactant concentration and could be arranged as the following order: PNP>PCP>PC>P. Moreover, in the case of phenolic compound separation, CPC achieved the highest treatment efficiency, and the separation efficiency of SDBS was a little lower than that of TX-100. The removal ratios of the same surfactant when treating different phenolic effluents were nearly similar. However, when treating the same phenolic compound, the sequence of the surfactant rejection was in the following order: TX-100>CPC>SDBS. These results indicate that CPC has a distinct superiority in the treatment of phenolic effluents via the MEUF process, and PNP easily solubilizes in the surface of the micelles.

Highly sensitive electrochemical sensor using a MWCNTs/GNPs-modified electrode for lead (II) detection based on Pb(2+)-induced G-rich DNA conformation.

A sensitive electrochemical lead ion (Pb(2+)) sensor based on carboxylic acid group functionalized multi-walled carbon nanotubes (MWNTs-COOH) and direct electrodeposited gold nanoparticles (GNPs) was developed for Pb(2+) detection. The DNA capture probe was self-assembled onto the surface of the modified electrode for hybridizing with the guanine-rich (G-rich) aptamer probe and for forming the DNA double helix structure. When Pb(2+) was added in, the DNA duplex unwound and formed a stabilized G-quadruplex (G4) due to the Pb(2+)-induced G-rich DNA conformation. Also, methylene blue (MB) was selected as the G4-binding indicator. Compared with previous Pb(2+) sensors, the proposed sensor had better sensitivity, because the modified MWCNTs/GNPs could provide a large surface area and good charge-transport capacity to dramatically improve the DNA attachment quantity and sensor performance. The sensor could detect Pb(2+) in a range from 5.0 × 10(-11) to 1.0 × 10(-14) M, with a detection of 4.3 × 10(-15) M.

Bioaccumulation of zinc, lead, copper, and cadmium from contaminated sediments by native plant species and Acrida cinerea in South China.

This research was conducted to search and identify spontaneously growing heavy metal-tolerant plant species that are potentially useful for phytoremediation in contaminated sediment. Five sites were selected for collection of plants growing on polluted shore (river bank) sediment of the Xiang River, China. The concentrations of Zn, Pb, Cu and Cd in plants, sediments, and grasshoppers were determined using flame atomic absorption spectrophotometer (AAS700, Perkin-Elmer, USA). Considering translocation factor and bioaccumulation factor, Rumex crispus (Polygonaceae), Rumex dentatus (Polygonaceae), and Lagopsis supina (Labiatae) could be potentially useful for phytostabilization of metals. R. crispus can be considered potentially useful for phytoextraction of Cd. In light of the biomagnification factors, grasshoppers are deconcentrators for Pb and Cd, microconcentrators for Zn and macroconcentrators for Cu to the plants, respectively. To the best of our knowledge, the present study is the first report on Zn, Pb, Cu and Cd accumulation in R. crispus and L. supina, providing a pioneer contribution to the very small volume of data available on the potential use of native plant species from contaminated sediments in phytostabilization and phytoremediation technologies.

Hydrolysis and acidification of waste-activated sludge in the presence of biosurfactant rhamnolipid: effect of pH.

In this investigation, the effect of pH (4.0-11.0) on waste-activated sludge (WAS) hydrolysis and acidification in the presence of a biosurfactant rhamnolipid (RL) were studied. The results showed that the hydrolysis and acidification of WAS in the presence of RL at alkaline pH values were more efficient than that at acidic and near-neutral pH values. After 6 h of hydrolysis, the soluble protein and carbohydrate were 1,654.7 and 675.9 mg/L (pH 11.0), and 825.6 and 376.0 mg/L (pH 7.0), whereas the values were only 315.0 and 84.0 mg/L at pH 4.0 and 164.1 and 32.0 mg/L for the blank, respectively. After 2 or 3 days of fermentation, the accumulated short-chain fatty acids (SCFAs) reached the highest and then decreased with a further increase in time at all investigated pH values. The analysis of SCFA compositions showed that acetic, propionic, and iso-valeric acids were the three main products at any pH value. A higher pH contributed to a greater proportion of acetic acid and a lesser proportion of iso-valeric acid; a lower pH resulted in a greater proportion of iso-valeric and lesser proportion of acetic acid in the initial fermentation. The proportions of acetic acid for the system with biosurfactant RL addition were 16.65, 36.33, and 62.94 %, respectively, at pH 4.0, 7.0, and 11.0 after 1 day. Correspondingly, the proportions were 40.34, 12.60, and 11.01 % for iso-valeric acid.

Microbial community analysis involved in the aerobic/extended-idle process performing biological phosphorus removal.

Recently, it has been found that biological phosphorus removal can be achieved in an aerobic/extended-idle (AEI) process using both glucose and acetate as the sole substrate. However, the microbial consortiums involved in glucose-fed and acetate-fed systems have not yet been characterized. Thus the aims of this paper were to investigate the diversities and dynamics of bacterial communities during the acclimation period, and to quantify polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs) in the systems. The phylogenetic analysis showed that the microbial communities were mainly composed of phylum Proteobacteria, Bacteroidetes, Chlorobi and another six kinds of unclassified bacteria. Fluorescence in-situ hybridization (FISH) analysis revealed that PAOs and GAOs accounted for 43 ± 7 and 16 ± 3% of all bacteria in the glucose-fed system, and 19 ± 4 and 35 ± 5% of total bacteria in the acetate-fed system, respectively. The results showed that the conventional PAOs could thrive in the AEI process, and a defined anaerobic zone was not necessarily required for putative PAOs growth.

Responses of Phanerochaete chrysosporium to toxic pollutants: physiological flux, oxidative stress, and detoxification.

The white-rot fungus Phanerochaete chrysosporium has been widely used for the treatment of waste streams containing heavy metals and toxic organic pollutants. The development of fungal-based treatment technologies requires detailed knowledge of the relationship between bulk water quality and the physiological responses of fungi. A noninvasive microtest technique was used to quantify real-time changes in proton, oxygen, and cadmium ion fluxes following the exposure of P. chrysosporium to environmental toxic (2,4-dichlorophenol and cadmium). Significant changes in H(+) and O(2) flux occurred after exposure to 10 mg/L 2,4-dichlorophenol and 0.1 mM cadmium. Cd(2+) flux decreased with time. Reactive oxygen species formation and antioxidant levels increased after cadmium treatment. Superoxide dismutase activity correlated well with malondialdehyde levels (r(2) = 0.964) at low cadmium concentrations. However, this correlation diminished and malondialdehyde levels significantly increased at the highest cadmium concentration tested. Real-time microscale signatures of H(+), O(2), and Cd(2+) fluxes coupled with oxidative stress analysis can improve our understanding of the physiological responses of P. chrysosporium to toxic pollutants and provide useful information for the development of fungal-based technologies to improve the treatment of wastes cocontaminated with heavy metals and organic pollutants.

Effect of dirhamnolipid on the removal of phenol catalyzed by laccase in aqueous solution.

In this study, the effects of three surfactants, i.e. the anionic biosurfactant dirhamnolipid (diRL), the cationic surfactant hexadecyltrimethyl ammonium bromide (CTAB), and the anionic surfactant sodium dodecyl sulfate (SDS), on the removal of phenol catalyzed by laccase were studied first. CTAB and SDS were detrimental, while diRL improved phenol removal and was selected for detailed research. The biosurfactant increased the activity of laccase and the removal of phenol with the increase of diRL concentrations from 10.6 to 318 µM. DiRL at 318 µM improved the removal when the initial concentrations of phenol were from 50 to 400 mg/l. In particular, the removal of phenol with 318 µM diRL was 4.3-6.4 folds that of the controls within 24 h when the initial concentration of phenol was 400 mg/l. The presence of diRL at 318 µM also caused the complete removal (above 98%) of phenol at concentrations from 50 to 400 mg/l after 24 h. The enhancement of phenol removal was over a wide range of pH and temperatures, and the highest removal efficiency was obtained at pH 6.0 and 50°C. The results suggest that diRL had potential application in the enhancement of phenols removal catalyzed by laccase in water treatment or remediation.

The isolation, identification of sludge-lysing thermophilic bacteria and its utilization in solubilization for excess sludge.

A novel strain of thermophilic bacteria with a highly efficient sludge dissolution performance was isolated from garden soil at 65 degrees C in this study. The colony morphology, physiological and biochemical characteristics of the strain were investigated. The results showed that the strain was Gram-positive, small rod-shaped, sporulating and secreted extracellular enzymes (protease and amylase). The 16S rDNA analysis demonstrated that this strain had not been previously reported. Therefore, it was labelled Bacillus thermophilic bacteria AT07-1 (registration number: FJ231108). To evaluate its capability for excess sludge solubilization, a pure culture of the strain was used in sludge solubilization tests; an enhanced solubilization process was subsequently obtained. After 36 h digestion, the protease activity in the inoculated system reached 0.37 U/ml, an increase of 0.16 U/ml compared with the non-inoculated system (0.21 U/ml). The solubilization rate for volatile suspended solids reached 46.45% in 48 h after inoculation with Bacillus thermophilic bacteria AT07-1, which was 10.24% higher than the non-inoculated system, and which could meet the standard of sludge stability suggested by the US Environmental Protection Agency.

A novel bifunctional europium chelate applied in quantitative determination of human immunoglobin G using time-resolved fluoroimmunoassay.

The authors demonstrate herein a novel time-resolved fluoroimmunoassay (TRFIA) protocol for quantification of human IgG with the new bifunctional chelate Eu(TTA)3(5-NH2-phen) (ETNP) labeling the goat anti-human IgG. The immunoassay was conducted by following the typical procedure for sandwich-type immunoreactions. Goat anti-human IgG was immobilized on aldehyde-modified glass slides. The human IgG analyte was first captured by the primary antibody and then sandwiched by a secondary antibody labeled with the chelate ETNP. The experimental procedure was simple to follow and gave desirable levels of sensitivity and low limits of detection. To the best of our knowledge, this is the first application of the new chelate, ETNP, in an immunoassay. In comparison to typical organic, fluorescent compounds and other lanthanide fluorescent chelates used in immunoassay, the detection sensitivity of our method using ETNP chelate in the solid phase was greatly improved and a concentration of human IgG about 5 µg/L could be detected under optimal conditions. The main result of this work shows that the new chelate ETNP can be applied as a powerful fluorescent labeling material for constructing ultrasensitive TRFIAs. The detection of human IgG, using ETNP as the chelate, is a model example of the effectiveness of this immunoassay. Many other types of antigen-antibody immunoassays should be possible using the protocol described herein.

An electrochemical DNA sensor based on a layers-film construction modified electrode.

This work developed a relatively inexpensive and layers-film construction electrochemical sensor for DNA recognition and its performance was investigated. The Fe(3)O(4) magnetic nanoparticles-cysteine were immobilized on the carbon paste electrode (CPE) surface using magnetic force. Multiwalled carbon nanotubes (MWCNTs), gold nanoparticles (GNPs), and chitosan (Chi) were used successively to coat on the electrode surface. The thiolated capture probe was assembled and competitively hybridized with the target nucleic acid and biotinylated response probe. The electrochemical behavior was analyzed by cyclic voltammetry and electrochemical impedance spectroscopy. In addition, the sensor performance was also analyzed by introducing the notion of detection efficiency. The experimental results showed that although the electron transfer capability of the CPE is less strong than that of a metal electrode used in the DNA sensor, the materials modified on the CPE could significantly improve the performance. A detection limit of 1 nM of target DNA and a sensitivity of 2.707 × 10(3) mA M(-1) cm(-2) were obtained. Although the resulting detection limit was not remarkable, further experiments could improve it.

Effects of continuous thermophilic composting (CTC) on bacterial community in the active composting process.

The method of continuous thermophilic composting (CTC) remarkably shortened the active composting cycle and enhanced the compost stability. Effects of CTC on the quantities of bacteria, with a comparison to the traditional composting (TC) method, were explored by plate count with incubation at 30, 40 and 50°C, respectively, and by quantitative PCR targeting the universal bacterial 16S rRNA genes and the Bacillus 16S rRNA genes. The comparison of cultivatable or uncultivatable bacterial numbers indicated that CTC might have increased the biomass of bacteria, especially Bacillus spp., during the composting. Denaturing gradient gel electrophoresis (DGGE) analysis was employed to investigate the effects of CTC on bacterial diversity, and a community dominated by fewer species was detected in a typical CTC run. The analysis of sequence and phylogeny based on DGGE indicated that the continuously high temperature had changed the structure of bacterial community and strengthened the mainstay role of the thermophilic and spore-forming Bacillus spp. in CTC run.

Mechanisms of efficient arsenite uptake by arsenic hyperaccumulator Pteris vittata.

Arsenate (AsV) and arsenite (AsIII) are two dominant arsenic species in the environment. While arsenate uptake is via phosphate transporter in plants, including arsenic hyperaccumulator Pteris vittata , AsIII uptake mechanisms by P. vittata are unclear. In this study, we investigated AsIII uptake by P. vittata involving root radial transport from external medium to cortical cells and xylem loading. In the root symplastic solution, AsIII was the predominant species (90-94%) and its concentrations were 1.6-21 times those in the medium. AsIII influx into root symplast followed Michaelis-Menten kinetics with K(m) of 77.7 µM at external AsIII concentrations of 2.6-650 µM. In the presence of metabolic inhibitor 2,4-dinitrophenol (DNP), arsenic concentrations in the root symplast were reduced to the levels lower than in the medium, indicating that a transporter-mediated active process was mainly responsible for AsIII influx into P. vittata roots. Unlike radial transport, AsIII loading into xylem involved both high- and low-affinity systems with K(m) of 8.8 µM and 70.4 µM, respectively. As indicated by the effect of 2,4-DNP, passive diffusion became more important in arsenic loading into xylem at higher external AsIII. The unique AsIII uptake system in P. vittata makes it a valuable model to understand the mechanisms of arsenic hyperaccumulation in the plant kingdom.

Effects of cake collapse caused by deposition of fractal aggregates on pressure drop during ceramic filtration.

A cake collapse model was developed by taking the combined effects of fractal dimension, relaxation ratio, coordination number, and aggregate diameter into consideration. The cake porosity including intraaggregate and interaggregate porosities was modeled successively by three typical coordination numbers (n = 6, 8, and 12). Accordingly, an inversion method made it possible to deduce the coordination number using the measured cake porosities, and the reverse-calculated value with minimum error and the corresponding relaxation ratios were applied as the parameters for the model. As a result, the profiles of intraaggregate and interaggregate porosities and cake porosity were respectively predicted in contrast to the integrated variation of the relaxation ratio and the fractal dimension. Furthermore, a comparison between the model predictions of the cake pressure drop gradients with and without aggregate compression was conducted to validate the presence of cake collapse. The results show that the predictions based on the proposed collapse model are in agreement with the experiments, and the coordination number is one of the key factors that must be incorporated into the cake collapse models.

A novel bifunctional europium complex as a potential fluorescent label for DNA detection.

A bifunctional europium complex of Eu(TTA)(3)(5-NH(2)-phen) using 2-thenoyltrifluoroacetonate (TTA) and 5-amino-1,10-phenanthroline (5-NH(2)-phen) as ligand reagents was applied in DNA detection assays for the first time. The complex has a long fluorescence lifetime, high fluorescence quantum yield, and is easy to label oligonucleotides for time-resolved fluorescence bioanalysis. A two-probe tandem DNA hybridization assay including capture DNA(1), probe DNA(2), and target DNA(3) was employed to detect microbial pathogens. The DNA sequences in the assay were designed using software Primer Premier 5.0 based on published specific nucleotide sequences of Staphylococcus aureus and Escherichia coli. 3'-Amino-modified capture DNA(1) was covalently immobilized on the common glass slide surface and the 5'-amino-modified probe DNA(2) was combined with the functionalized Eu(TTA)(3)(5-NH(2)-phen) via glutaraldehyde. The detection was done by monitoring the fluorescence intensity from the glass surface after the hybridization reaction with complementary target DNA(3). The optimal concentration of capture DNA(1) of 1.0 x 10(-6) mol l(-1) dropped onto the glass slides and optimal hybridization temperatures of 48 degrees C and 39 degrees C respectively for Staphylococcus aureus and Escherichia coli were obtained. The proposed DNA detection system showed higher sensitivity than such a complex doped nanoparticle-based detection system in our previous study for the better uniformity and dispersity of monomolecular labels. The sensing system presented a short hybridization time of 2 h, satisfactory stability, and high selectivity. The results demonstrate that this complex might be a potentially excellent dye in area of biochemical analysis.