Toward nitrogen recovery: Co-cultivation of microalgae and bacteria enhances the production of high-value nitrogen-rich cyanophycin.

The algal-bacterial wastewater treatment process has been proven to be highly efficient in removing nutrients and recovering nitrogen (N). However, the recovery of the valuable N-rich biopolymer, cyanophycin, remains limited. This research explored the synthesis mechanism and recovery potential of cyanophycin within two algal-bacterial symbiotic reactors. The findings reveal that the synergy between algae and bacteria enhances the removal of N and phosphorus. The crude contents of cyanophycin in the algal-bacterial consortia reached 115 and 124 mg/g of mixed liquor suspended solids (MLSS), respectively, showing an increase of 11.7 %-20.4 % (p < 0.001) compared with conventional activated sludge. Among the 170 metagenome-assembled genomes (MAGs) analyzed, 50 were capable of synthesizing cyanophycin, indicating that cyanophycin producers are common in algal-bacterial systems. The compositions of cyanophycin producers in the two algal-bacterial reactors were affected by different lighting initiation time. The study identified two intracellular synthesis pathways for cyanophycin. Approximately 36 MAGs can synthesize cyanophycin de novo using ammonium and glucose, while the remaining 14 MAGs require exogenous arginine for production. Notably, several MAGs with high abundance are capable of assimilating both nitrate and ammonium into cyanophycin, demonstrating a robust N utilization capability. This research also marks the first identification of potential horizontal gene transfer of the cyanophycin synthase encoding gene (cphA) within the wastewater microbial community. This suggests that the spread of cphA could expand the population of cyanophycin producers. The study offers new insights into recycling the high-value N-rich biopolymer cyanophycin, contributing to the advancement of wastewater resource utilization.

Amoxicillin degradation and high-value extracellular polymer recovery by algal-bacterial symbiosis systems.

Algal-bacterial symbiosis systems have emerged as sustainable methods for the treatment of new pollutants and the recovery of resources. However, the bio-refinery of biomass derived from microalgae is inefficient and expensive. In order to simultaneously degrade antibiotic and recover resources efficiently, two algal-bacterial symbiosis systems were constructed using Pseudomonas aeruginosa (alginate overproduction) and Bacillus subtilis (poly-γ-glutamic acid overproduction) with amoxicillin-degrading-microalga Prototheca zopfii W1. The optimal conditions for W1 to degrade amoxicillin are 35 °C, pH 7, and 180 rpm. In the presence of 5-50 mg/L of amoxicillin, W1-P. aeruginosa and W1-B. subtilis exhibit higher amoxicillin degradation and produce more extracellular polymers than W1 or bacteria alone. The metabolomic analysis demonstrates that the algal-bacterial symbiosis enhances the tolerance of W1 to amoxicillin by altering carbohydrate metabolism and promotes the production of biopolymers by upregulating the precursors synthesis. Moreover, the removal of amoxicillin (10 mg/L) from livestock effluent by W1-P. aeruginosa and W1-B. subtilis is greater than 90 % in 3 days, and the maximum yields of alginate and poly-γ-glutamate are 446.1 and 254.3 mg/g dry cell weight, respectively. These outcomes provide theoretical support for the application of algal-bacterial symbiosis systems to treatment of amoxicillin wastewater and efficient production of biopolymers.

Transition metal-doped germanium oxide nanozyme with enhanced enzyme-like activity for rapid detection of pesticide residues in water samples.

Designing highly active nanozymes for bioanalysis and environmental sensing remains a challenge. In this study, transition metal, palladium (Pd) and iron (Fe), doped germanium oxide (GeO2) nanozyme was designed and optimized. Compared with the pristine GeO2 nanozyme, the transition metal doped GeO2 nanozyme have lower Michaelis-Menten constants and higher catalytic activity, indicating that the Pd and Fe doped GeO2 nanozyme not only enhance their affinity for the substrate but also improve its catalytic activity. In addition, a colorimetric sensor based on the GeO2@Pd-H2O2-TMB system was constructed for the visual detection of simazine in water samples due to the good affinity between TMB and simazine. This sensor has good selectivity and sensitivity with a detection limit of 6.21 µM because of the highest catalytic performance of GeO2@Pd nanozyme. This study broadens the application of nanozymes in environmental field and other nanozymes can also be enhanced in activity by simple transition metal doping.

Efficient and recyclable degradation of organic dye pollutants by CeO2@ZIF-8 nanozyme-based non-photocatalytic system.

Advanced oxidation processes-based catalysis system as the most typical pollutant degradation technology always suffer from poor durability and photo-dependent. Inspired by the fact that some nanomaterials exhibit catalytic properties closer to natural enzymes, a high peroxidase-like activity and stability CeO2@ZIF-8 nanozyme was synthesized in this study for non-photodegradation of dyes pollution. Multiple characterization techniques were applied to prove the successful synthesis of the nanozyme. The influence of different parameters on the catalytic degradation of organic dye by nanozyme was investigated. This nanozyme achieved a maximum degradation efficiency of 99.81% for methyl orange and maintained its catalytic performance in repeated experiments. Possible degradation intermediates and pathways for methyl orange were then proposed. In addition, the CeO2@ZIF-8 loaded starch/agarose films were prepared for the portable and recyclable remediation of real dye wastewater, which maintained more than 80% degradation efficiency after 5 successive cycles. These results suggested that nanozyme based non-photocatalytic system is a potential catalyst for dye degradation and it opens a new avenue to develop high-performance and recyclable catalysts for pollutant remediation.

Alginate production of Pseudomonas strains and its application in preparation of alginate-biomass hydrogel for heavy metal adsorption.

In this study, 7 Pseudomonas strains were isolated from a wastewater treatment plant, and the alginate production of Pseudomonas strains under different environmental conditions was evaluated. Subsequently, alginate-biomass hydrogel beads were prepared using alginate and biomass of Pseudomonas, and their adsorption performances and mechanism to Pb2+ and Cd2+ were analyzed. The results show that weakly acidic pH and 37 °C is favorable for alginate synthesis of Pseudomonas strains, and P. alcaligenes YLS18 have the highest alginate yield (29.4 mg/g). The adsorption processes of Pb2+ and Cd2+ by hydrogel beads are well described by Langmuir model, indicating that the adsorption process is monolayer. Among the biomass of these strains, P. nitroreducens YLB32 shows the highest biosorption capacities, reaching 110.7 mg/g for Pb2+ and 54.3 mg/g for Cd2+ at pH 5. Alginate-biomass hydrogel beads obtain higher adsorption capacity to Pb2+ (184.0 mg/g) and Cd2+ (92.4 mg/g), and exhibit good reusability. The adsorption mechanism of Pb2+ and Cd2+ by hydrogel beads involves physical tapping of ions, electrostatic interactions, complexation, cation exchange and precipitation. These results provide strong support for promoting alginate recovery from activated sludge and for treating heavy metal wastewater.

Cyanophycin Granule Polypeptide: a Neglected High Value-Added Biopolymer, Synthesized in Activated Sludge on a Large Scale.

Recovery of microbial synthetic polymers with high economic value and market demand in activated sludge has attracted extensive attention. This work analyzed the synthesis of cyanophycin granule peptide (CGP) in activated sludge and its adsorption capacity for heavy metals and dyes. The distribution and expression of synthetic genes for eight biopolymers in two wastewater treatment plants (WWTPs) were analyzed by metagenomics and metatranscriptomics. The results indicate that the abundance and expression level of CGP synthase (cphA) are similar to those of polyhydroxyalkanoate polymerase, implying high synthesis of CGP in activated sludges. CGP in activated sludge is mainly polymerized from aspartic acid and arginine, and its secondary structure is mainly ß-sheet. The crude yields of CGP are as high as 104 ± 26 and 76 ± 13 mg/g dry sludge in winter and in summer, respectively, comparable to those of polyhydroxyalkanoate and alginate. CGP has a stronger adsorption capacity for anionic pollutants (Cr (VI) and methyl orange) than for cationic pollutants because it is rich in guanidine groups. This study highlights prospects for recovery and application of CGP from WWTPs. IMPORTANCE The conversion of organic pollutants into bioresources by activated sludge can reduce the carbon dioxide emission of wastewater treatment plants. Identification of new high value-added biopolymers produced by activated sludge is beneficial to recover bioresources. Cyanophycin granule polypeptide (CGP), first discovered in cyanobacteria, has unique chemical and material properties suitable for industrial food, medicine, cosmetics, water treatment, and agriculture applications. Here, we revealed for the first time that activated sludge has a remarkable ability to produce CGP. These findings could further facilitate the conversion of wastewater treatment plants into resource recycling plants.

Metagenomics and metatranscriptomics analyses of antibiotic synthesis in activated sludge.

The generic of antibiotics is considered to be a main reason for the generation of antibiotic resistance genes (ARGs) in wastewater treatment plants (WWTPs). However, little has been reported about the antibiotic biosynthesis by activated sludge. In this study, the distribution and expression of antibiotic biosynthetic genes (ABGs) in the floc sludge and biofilm from two WWTPs were deciphered using metagenomics and metatranscriptomics. The results showed that 2% of the community were in general well-linked to antibiotic production, indicating a non-negligible antibiotic synthetic ability of WWTPs. 93 ABGs belonging to 26 antibiotics were determined, among which aminoglycosides, ß-lactams, ansamycins, peptides, macrolides were majority. The relative abundances of detected ABGs had a large interval, ranging from 0.000006% to 0.042%. The predominant antibiotic types of synthetic genes with higher relative expression levels were monobactams, penicillin & cephalosporins and streptomycin, primarily belonging to ß-lactams and aminoglycosides. The hypothetical synthetic pathways of streptomycin synthesis and penicillin & cephalosporin synthesis were proposed. And the coexistence of ABGs and ARGs for these two antibiotics was also pronounced in activated sludge from meta-omics data. These findings for the first time demonstrated the antibiotic synthetic potential in activated sludges, revealing new sources of antibiotics and resistance genes in WWTPs, and thereby aggravating environmental pollution.

Quantitative proteomics and phosphoproteomics elucidate the molecular mechanism of nanostructured TiO2-stimulated biofilm formation.

With the increasing concerns regarding bacterial adaption to nanomaterials, it is critical to explore the main mechanism behind the adaptive morphogenesis of microorganisms. In this work, the biofilms formed from activated sludge exposed to 5 and 50 mg/L nTiO2 in the dark had increased biomass and selectively enriched pathogens. To further elaborate adaptive mechanism of biofilm formation induced by nTiO2, the protein response and protein phosphorylation modification of Escherichia coli K12 were determined using integrative systems biology analyses of proteomics and phosphoproteomics. Results identified that E. coli cultivated with nTiO2 considerably upregulated iron acquisition, and regulated protein phosphorylation states associated with of transcription and translation and biofilm formation relative to unexposed controls. Accordingly, bacteria increased siderophores and exopolysaccharide content (increased by about 57% and 231%, respectively), and enhanced resistance to transcriptional inhibitory antibiotics. Moreover, a dose of an iron chelator, i.e., deferoxamine mesylate salt, effectively retarded the biofilm development of bacteria exposed to 50 mg/L nTiO2. Overall, this work will provide a new insight for biofouling control, and contribute to an improved understanding of microbial adaption to nanomaterials.

A persistent luminescent nanobeacon for practical detection of lead ions via avoiding background interference.

Heavy metal ions are considered to be the most serious sources for water pollution. Accurate detection of metal ions is important for pollution control and ecological protection. Background interference is an inevitable obstacle in the fluorescent analysis of complex samples. Herein, a persistent luminescent nanobeacon is communicated to detect metal ions in real samples via avoiding background interference. The nanobeacon is constituted by persistent luminescence nanomaterials and metal-specific DNAzymes. As a proof of concept, Zn2GeO4: Mn persistent luminescence nanorods (PLNRs) was synthesized and functionalized with the 17E DNAzyme for lead ion (Pb2+) detection. As a result, in the luminescent manner, the nanobeacon could recognize Pb2+ selectively and detect it with high signal-to-background ratios (SBR) both in buffer and real samples, but the fluorescent SBR declined significantly when used in real samples. Thus, this persistent luminescent nanobeacon can achieve practical detection of metal ions via avoiding background interference. Compared to previous methods of improving signal-to-background ratio, this persistent luminescent nanobeacon is more accessible, and all DNAzyme-specific ions can be directly adapted.

Copper ion and G-quadruplex-mediated fluorescent sensor for highly selective detection of bleomycin in actual samples.

Improper dosage of Bleomycin (BLM) can easily lead to a series of side effects such as pulmonary fibrosis and pulmonary toxicity. Therefore, detecting the content of BLM in actual sample is very helpful to make full use of its therapeutic efficacy and reduce its toxicity. Herein, we constructed a copper ion and G-quadruplex mediated label-free sensor to detect BLM. The strategy mainly relies on the chelation of BLM to copper ions, which makes the copper ions lose the quenching ability to the fluorescent dye N-methylmesoporphyrin (NMM) after chelation. With the assistance of the G-quadruplex, the BLM content in the sample can be detected by observing the change in fluorescence. A good linear relationship can be clearly observed within the BLM concentration range of 0.1 nM-75 nM, and the limit of detection was derived as 0.1 nM. This sensor did not involve any labeling or addition of Fe2+, even in the presence of 10 different antibiotics and 11 different metal ions, it still has a good monitoring effect, and can be successfully applied to the detection of BLM in serum and wastewater. Thus, we believe that this work hold great potential in antibiotic monitoring and environmental protection.

Transformation and kinetics of chlorine-containing products during pyrolysis of plastic wastes.

Pyrolysis can not only effectively dispose of plastic wastes but also reclaim valuable chemicals and biochar. However, the production and release of second pollutants, particularly chlorine-containing products, have been neglected. The mechanism for the transformation of chlorine during the pyrolysis of plastic wastes remains unclear. Herein, a thermogravimetric Fourier transform infrared mass spectrometry technology was used to investigate the migration and transformation of substances during the pyrolysis of polyvinyl chloride (PVC) plastic from 200 °C to 900 °C with heating rates of 5, 50, 100, 150, and 200 K min-1. Results show the first stage of weight loss is at 200 °C-360 °C, where the dehydrochlorination of PVC mainly occurred, accompanied by the formation of conjugated double bonds and a small number of hydrocarbon compounds. The second stage of weight loss is at 360 °C-550 °C, where the breakage and rearrangement of the long polyethene chain may occur. Kinetics analysis shows the higher activation energy value is in the second stage, which indicates that the second stage reaction is less likely to occur and the Flynn-Wall-Ozawa method is more suitable for the study of plastic pyrolysis kinetics. This study suggests that second pollutants can be minimized during controllable pyrolysis.

Adsorption-improved MoSe2 nanosheet by heteroatom doping and its application for simultaneous detection and removal of mercury (II).

Water pollution arising from heavy metal ions continues to be a major environmental problem, which represents a serious threat to human beings and animals worldwide. New materials that can simultaneously detect and remove these toxic ions are urgently required. Herein, nitrogen and sulfur co-doped molybdenum selenide nanosheets (N, S-MoSe2) were prepared and found to be fluorescently responsive to mercury (II) with an improved adsorption capacity (208.33 mg g-1), thereby providing the possibility for the simultaneous detection and removal of mercury (II) in water samples. The great affinity was the result of the complexation of mercury (II) with Se and S atoms in N, S-MoSe2 as well as the electrostatic adsorption of cation mercury (II) on negatively charged N, S-MoSe2. Besides good sensitivity and selectivity toward mercury (II), N, S-MoSe2 displayed a relatively consistent performance under a wide pH range from 3 to 10. The removal efficiency reached 87.5% with fast adsorption kinetics, and N, S-MoSe2 could be reused after simple treatment. Thus, this work is expected to provide new material for the detection and removal of mercury (II) in an aqueous solution and offer an insight into the interaction between heavy metal ions and inorganic nanomaterials.

Graphene oxide-regulated low-background aptasensor for the "turn on" detection of tetracycline.

Tetracyclines (TC) are a common antibiotic for using in livestock breeding and healthcare; however, due to the inappropriate application of TCs, more than 75% of TCs are excreted and released into the environment in an active form through human and animal urine and feces, which results in high levels of TCs in the ecological system, causing adverse effects on the food safety and human health. Thus, the high-performance monitoring of TC pollution is necessary. In this work, a highly sensitive fluorescent aptasensor was developed that was based on graphene oxide (GO) regulation of low background signal and target-induced fluorescence restoration. In the absence of analyte, the DNA probe (TC aptamer) was adsorbed completely by GO and failed to enhance the fluorescence of SYBR gold (SG), thereby resulting in a low background signal. When the TC-included samples were added, the DNA probe formed an aptamer-TC complex, thereby separating from the surface of the GO and inducing the fluorescence of SG. Under optimal conditions, the proposed strategy could detect TC concentrations of less than 6.2 × 10-3 ng mL-1, which is four orders of magnitude better than the detection limit of the "turn off" mode (53.9511 ng mL-1). Moreover, this aptasensor has been used to detect TC from milk samples and wastewater samples, and its satisfactory performances demonstrate that the proposed strategy can be applied in practice for TC monitor in food safety and environmental protection. Therefore, we believe that this work is meaningful in pollution monitoring, environment restoration and emergency treatment.

Fluorescent and colorimetric dual-mode detection of tetracycline in wastewater based on heteroatoms-doped reduced state carbon dots.

A large amount of tetracycline (TC) persists in water, soil, food, and feed due to the overuse of antibiotics, causing serious environmental problems such as damage to ecosystems and posing risks to human health. Thus, there is an urgent need to find a method to detect TC that is practical, rapid, sensitive, and offers ready visualization of TC levels so that adequate remediation measures can be immediately implemented. Herein, we report a fluorescent and colorimetric dual-mode nanosensor for the detection of TC based on reduced state carbon dots (r-CDs). In the presence of TC, the emission fluorescence of r-CDs was quenched by the Förster resonance energy transfer mechanism to achieve high-sensitivity detection of TC with a low limit of detection (LOD) of 1.73 nM. Moreover, TC was also detected by r-CDs via a noticeable color change of the solution (from colorless to red) with the naked eye, having an LOD of 0.46 µM. Furthermore, r-CDs have excellent selectivity and sensitivity in detecting TC in wastewater, and therefore, have practical applications in wastewater treatment. The fluorescent and colorimetric dual-mode proposed in this work may offer a unique idea for the detection of TC, with great prospects for environmental wastewater applications.

Fluorometric determination of the breast cancer 1 gene based on the target-induced conformational change of a DNA template for copper nanoclusters.

The breast cancer 1 (BRCA1) gene is a tumor suppressor gene, whose mutation is closely related to breast cancer. Therefore, the sensitive detection of the BRCA1 gene is extremely important for human health, particularly for women. In this study, a label-free fluorescent method based on hairpin DNA-templated copper nanoclusters (CuNCs) was for the first time developed for the detection of the BRCA1 gene. In the absence of target DNA, the detection system showed a strong red emission and produced a high emission peak. However, in the presence of the BRCA1 gene, the DNA probe hybridized with the BRCA1 gene and conformation of the DNA probe changed. As a result, the amount of produced CuNCs decreased and a low emission peak was obtained. The fluorescence intensity of the detection system was linearly correlated with the concentration of the BRCA1 gene ranging from 2 nM to 600 nM. The detectable limit was 2 nM for the BRCA1 gene assay, which was comparable with those reported by other non-amplifying sensors. Moreover, the developed method showed satisfactory recoveries for the BRCA1 gene assay in the bovine serum. The DNA-templated CuNC-based fluorescent assay thus offered a promising platform for the diagnosis of a breast cancer biomarker.

Nanoparticles-EPS corona increases the accumulation of heavy metals and biotoxicity of nanoparticles.

Microbial extracellular polymeric substances (EPS) coating nanoparticles (NPs) surface can form NPs-EPS corona, which significantly affect the adsorption of NPs to toxic substances and alter the ecotoxicological effect of NPs. In this work, the EPS coronas on TiO2 NPs (TNPs) and CeO2 NPs (CNPs) were characterized and the adsorption characteristics of NPs with and without EPS corona to five heavy metals were investigated in single-metal and multiple-metal systems. The results of spectral analysis showed that NPs-EPS corona exhibited new crystalline phases and abundant functional groups. Moreover, 42 and 13 proteins were identified in the TNPs-EPS and CNPs-EPS coronas, respectively. The rates of Cd2+, Pb2+, Cu2+, Ni2+ and Ag+ adsorption by NPs-EPS corona increased to values that were 6.7-7.6, 4.4-5.1, 4.2-5.5, 3.9-4.9 and 8.5-8.8 times those of NPs without EPS corona, respectively, in single-metal system. NPs-EPS coronas are effective in absorbing Ag+, Pb2+ and Cu2+ compared with Cd2+and Ni2+ in multiple metal adsorption. These results indicated that NPs-EPS corona effectively adsorb and remove heavy metals by forming NPs-EPS-metal complexes and inducing precipitation. However, NPs-EPS corona can enhance the toxicity of NPs by accumulating highly-toxic heavy metals in aquatic environments.

Graphene biosensors for bacterial and viral pathogens.

The infection and spread of pathogens (e.g., COVID-19) pose an enormous threat to the safety of human beings and animals all over the world. The rapid and accurate monitoring and determination of pathogens are of great significance to clinical diagnosis, food safety and environmental evaluation. In recent years, with the evolution of nanotechnology, nano-sized graphene and graphene derivatives have been frequently introduced into the construction of biosensors due to their unique physicochemical properties and biocompatibility. The combination of biomolecules with specific recognition capabilities and graphene materials provides a promising strategy to construct more stable and sensitive biosensors for the detection of pathogens. This review tracks the development of graphene biosensors for the detection of bacterial and viral pathogens, mainly including the preparation of graphene biosensors and their working mechanism. The challenges involved in this field have been discussed, and the perspective for further development has been put forward, aiming to promote the development of pathogens sensing and the contribution to epidemic prevention.

Beyond native deoxyribonucleic acid, templating fluorescent nanomaterials for bioanalytical applications: A review.

Due to its unique programmability, nanosized structure, and biocompatible properties, deoxyribonucleic acid (DNA) has attracted increasing attention for the construction of versatile nanostructures and nanomaterials. This review summarizes the recent developments in DNA-templated fluorescent nanomaterials, including DNA-stabilized quantum dots (QDs) and DNA-templated metal nanoclusters (NCs), as well as their applications in bioanalysis. These fluorescent nanomaterials not only have good fluorescence properties but also exhibit excellent performance in DNA recognition, which greatly expands the range of their bioanalytical applications. Finally, we discuss some current challenges and future work in this field, with the goal of further promoting the potential applications and developments of DNA-templated fluorescent nanomaterials in biochemical analysis.

Recent progress in copper nanocluster-based fluorescent probing: a review.

Copper nanoclusters (CuNCs) are an attractive alternative to other metal nanoclusters. The synthesis of CuNCs is highly efficient and fast, with low-cost and without any complicated manipulation. Because of their tunable fluorescence and low toxicity, CuNCs have been highly exploited for biochemical sensing. This review (with 172 refs.) summarizes the progress that has been made in the field in the past years. Following an introduction into the fundamentals of CuNCs, the review first focuses on synthetic methods and the fluorescence properties of CuNCs (with subsections on the use of proteins, peptides, DNA and other molecules as templates). This is followed by a section on the use of CuNCs in fluorometric assays, with subsections on the detection of small molecules, proteins, nucleic acids, various other biomolecules including drugs, and of pH values. A further large chapter summarizes the work related to environmental analyses, specifically on determination of metal ions, anions and pollutants. Graphical abstract Schematic representation of the synthesis and potential applications of copper nanocluster (CuNCs) in biochemical analysis, emphatically reflected in some vital areas such as small molecule analysis, biomacromolecule monitoring, cell imaging, ions detection, toxic pollutant, etc.

High specific MNase assay for rapid identification of Staphylococcus aureus using AT-rich dsDNA substrate.

Micrococcal nuclease (MNase) is a nonspecific endo-exonuclease that digests single-stranded/double-stranded DNA and RNA. The existence of MNase can serves as an important diagnostic biomarker of Staphylococcus aureus (S. aureus) infection. However, most of the substrates in MNase-based sensors are single-stranded DNA, which could also be digested by exonuclease I or S1 nuclease and interfere the MNase detection. In this work, we developed a highly selective fluorescent method for MNase detection using a specific dsDNA and nucleic acid dye SYBR Green I (SGI) as the indicator. After rational design, an AT-rich dsDNA with 3' protruding termini was screened as the high-specific substrate of MNase assay and efficient enhancer of SGI. The AT-rich dsDNA substrate can resist the digestion of other exonuclease and greatly enhance the fluorescence of SGI. This high-specific substrate-based probe can detect MNase in buffer as well as biological sample with highly selectivity. Moreover, this method was also applied to monitor the MNase secreted by S. aureus. Thus, the proposed MNase-based assay has a strong potential to identify S. aureus in food safety and microbial infection due to its excellent analytical sensitivity and high selectivity.