Protein-like Fluorophores Research Articles

Characterization of metal-binding behavior of DOM component structure in biochar: Analysis of binding sites, binding sequences and impact pathways

The binding of biochar-derived dissolved organic matter (BDOM) to heavy metals is a complex and dynamic process, with potentially different impact pathways. The outer electrons of various heavy metals can also affect their binding ability. This study comprehensively investigated the binding sites, sequences, constants, and influencing pathways of each component structure in BDOM through modern spectroscopic methods and multidimensional chemometrics. Here, the dominance of carboxyl and phenolic groups in the interaction of chromogenic dissolved organic matter (CDOM) with metal ions, which complex with BDOM, leads to an increase in the size of the BDOM molecule. In addition, Cu and Zn primarily bound to humic acids fluorophore, while Pb and Ni preferentially bound to protein-like fluorophore, likely due to the varying affinities of nitrogen- and carbon-based functional groups for different metals. Significant differences were observed in the interactions between different metal ions and fluorophores, involving both direct and indirect effects. Molecular perspectives and pathways suggested that the binding of BDOM to Zn was notably intricate, involving multiple conformational changes and impact pathways. These findings offer a crucial scientific foundation for a comprehensive comprehension of the interaction mechanisms between various metal ions and BDOM components.

Response of a source water quality through a heavy precipitation event: Nutrients, dissolved organic matter and their DBPs formation

To better understand the influence of heavy precipitation event on water supply security, the change of nutrients, dissolved organic matter (DOM) and their corresponding disinfection by-products (DBPs) generation in river were investigated through a heavy precipitation event. The results show that short-term sustained heavy precipitation causes additional load on the river. Significant temporal variations and increases in dissolved oxygen (DO), turbidity, dissolved organic carbon (DOC) and molecular weight of raw water were observed during the heavy precipitation, and dissolved organic nitrogen (DON) showed a remarkably decline with the continuation of precipitation. The molecular weight and aromaticity of DOM increased dramatically with the continuous superposition of precipitation and flow, and the proportion of hydrophobic fraction increased during the study period. Four different fluorescent components were identified, including humic-like (C1, C2 and C3), and protein-like fluorophore C4. Heavy precipitation events affected the mobilization of DOM, while the generation of DBPs increased by 4%–47% during the chlorination, and the elevated river level further increased the DBPs formation. In particular, the HOA fraction was the main contributor of DBPs (up to 70%). The risk of haloacetic acids (HAAs) generation caused by heavy rainfall was concentrated in the river runoff producing and regression periods for both hydrophobic and hydrophilic fractions of DOM. Humification index (HIX) was positively correlated with the yield level of TCM, DCAA, TCAA, and DCAN during chlorination. Chlorination disinfection of raw water during river runoff producing and regression period is likely to increase the haloacetonitriles formation potential (HANsFP), thus posing a vital threat to water supply safety, while chloramination disinfection may significantly relieve the threat. This study provides a theoretical basis for the direct/indirect link between short-term precipitation events and their impact on water quality.

Functional amyloid fibrils of biofilm-forming marine bacterium Pseudomonas aeruginosa PFL-P1 interact spontaneously with pyrene and augment the biodegradation

Bacteria thrive in biofilms embedding in the three-dimensional extracellular polymeric substances (EPS). Functional Amyloid in Pseudomonas (Fap), a protein in EPS, efficiently sequesters polycyclic aromatic hydrocarbons (PAHs). Present study reports the characterization of Fap fibrils from Pseudomonas aeruginosa PFL-P1 and describes the interaction with pyrene to assess the impact on pyrene degradation. Overexpression of fap in E. coli BL21(DE3) cells significantly enhances biofilm formation (p < 0.0001) and amyloid production (p = 0.0002), particularly with pyrene. Defibrillated Fap analysis reveals FapC monomers and increased fibrillation with pyrene. Circular Dichroism (CD), Fourier Transform Infrared Spectroscopy (FTIR), and X-ray Diffraction (XRD) unveil characteristic amyloid peaks and structural changes in Fap fibrils upon pyrene exposure. 3D-EEM analysis identifies a protein-like fluorophore in Fap fibrils, exhibiting pyrene-induced fluorescence quenching. Binding constants range from 5.23 to 7.78 M−1, with ΔG of −5.10 kJ mol−1 at 298 K, indicating spontaneous and exothermic interaction driven by hydrophobic forces. Exogenous Fap fibrils substantially increased the biofilm growth and pyrene degradation by P. aeruginosa PFL-P1 from 46 % to 64 % within 7 days (p = 0.0236). GC–MS identifies diverse metabolites, implying phthalic acid pathway in pyrene degradation. This study deepens insights into structural dynamics of Fap fibrils when exposed to pyrene, offering potential application in environmental bioremediation.

Compositional and optical characteristics of aqueous brown carbon and HULIS in the eastern Indo-Gangetic Plain using a coupled EEM PARAFAC, FT-IR and 1H NMR approach

This study provides insights into the fluorophoric composition of aqueous brown carbon (BrCaq) and chemically-separated humic-like substances (HULIS): neutral HULIS (HULIS-n; at pH = 7) and acidic HULIS (HULIS-a; at pH = 2) on a seasonal and day-night basis in the eastern Indo-Gangetic Plain (IGP), India. A coupled approach including excitation-emission matrix (EEM) fluorescence and parallel factor analysis (PARAFAC) model, Fourier-transformed infrared spectroscopy (FT-IR) and proton nuclear magnetic resonance (1H NMR) spectroscopy was employed to understand the links between structural, compositional and fluorophoric characteristics of BrCaq and HULIS fractions. HULIS fluorophores (HULISfluoro) with varying oxidation states transported from the northwest IGP were dominant during biomass burning seasons (post-monsoon and winter), while protein-like fluorophores (PRLISfluoro) from marine emissions showed large contributions during summer. HULIS-n moieties were mostly primary in nature with higher conjugation, while HULIS-a were associated with secondarily formed and aged species with a larger contribution from degradation products. A substantial presence of tyrosine-like proteins in both chemically-separated HULIS fractions indicated that atmospheric HULIS is not entirely humic or fulvic-like in the eastern IGP. Finally, the dominance of H-C-O groups across seasons suggested consistent fossil fuel signatures along with season-specific influence of photodegradable cellulose from marine organisms in the summer and biomass burning in the post-monsoon and winter.

Investigating the molecular weight distribution of atmospheric water-soluble brown carbon using high-performance size exclusion chromatography coupled with diode array and fluorescence detectors

Atmospheric brown carbon (BrC) contain amounts of organic species, but their molecular weight (MW) distributions is still poorly understood. This study applied high-performance size exclusion chromatography (HPSEC) coupled with a diode array detector (DAD) and fluorescence detector (FLD) to characterize the MW distributions of typical chromophores and fluorophores within water-soluble BrC. The investigation focused on the spring season, encompassing both typical urban and rural aerosols. Our results showed that chromophores (at 254 and 365 nm), and humic-like and protein-like fluorophores identified by excitation-emission matrix parallel factor analysis (EEM-PARAFAC) within BrC were broadly distributed along the MW continuum (∼50–20,000 Da). This suggests that BrC mainly comprises complex chromophores and fluorophores with heterogeneous molecular sizes. High-MW (HMW, >1 kDa) species (66%–74%) dominated the chromophores at 254 and 365 nm. However, the latter chromophores were enriched with more HMW species. This result suggested that the HMW chromophores might contribute more to BrC absorption at longer wavelengths. The PARAFAC-derived fluorescent components also exhibited different MW distributions. Three humic-like substances (HULIS) were all dominated by HMW fractions (51%–74%), but protein-like fluorescent component (PLOM) enriched low-MW (LMW, <1 kDa) species (60%–66%). Furthermore, the molecular size (i.e., weight-averaged and number-averaged MW) and the ratios between HMW and LMW species decreased in the order highly-oxygenated HULIS > less-oxygenated HULIS > PLOM, indicating that the fluorophores with longer Em were generally related to larger MW. To our knowledge, this is the first report on the molecular size of individual fluorescent components within aerosol BrC. The results obtained here enhanced our knowledge of heterogeneous composition, complex physicochemical properties, and potential atmospheric fates of aerosol BrC.

Source characterization of dissolved organic matter in the eastern Beagle Channel from a spring situation

Dissolved organic matter (DOM) was analyzed by its optical and fluorescent properties in order to study its distribution, source and transformation in relation to microbial abundances, chlorophyll and physicochemical gradients along a transect in the eastern Beagle Channel (BC) during austral spring 2019. Moreover, DOM evolution was followed during three tidal cycles in two fixed stations (F1, F2) separated by the Mackinlay Strait, a hydrographic frontier between water masses under stronger continental influence (inner sector) and that of less modified Sub-Antarctic waters (outer sector). The fluorescence signals of protein- and humic-like compounds were used as a proxy for labile and non-labile material, respectively, while the concentration of nano-, pico- and virioplankton populations, chlorophyll-a (Chl-a) and dissolved organic carbon (DOC) were used as a proxy for biological activity. Moreover, several spectroscopic indices, such as the indicator of microbially produced DOM (BIX), the Freshness index (FIX), an indicator of molecular weight, the Slope Ratio (SR) and an indicator of chromophoric DOM (CDOM), were used to trace the quality and origin of DOM. Chl-a concentration was generally low in the sampling area (< 1 μg L−1) but increased towards the end of the campaign and especially during the visit of F1 along with the concentration of picoeukaryotes and the protein-like fluorophores evidencing the onset of a bloom event. Protein-like fluorophores peaked in the euphotic zone of the inner sector of the BC while humic-like material of terrestrial origin was homogeneously distributed throughout the water column and showed a decreasing gradient towards the outer sector and Atlantic waters. Furthermore, the inner sector was characterized by higher amounts of CDOM, high molecular weight material and more recalcitrant DOM. Accumulation of autochthonous humic-like material in bottom waters of the inner sector along with enhanced values of FIX and BIX preceded by colored organic material suggest an efficient functioning of the microbial carbon pump fueled by terrestrial DOM sources in these waters.

Insights into dissolved organics in non-urban areas - Optical properties and sources

Brown carbon aerosols show obvious light absorption properties in the ultraviolet–visible (UV–Vis) range, which has an important impact on photochemistry and climate. In this study, experimental samples originated from the North slope of the Qinling Mountains (at two remote suburb sites) to study the optical properties of water-soluble brown carbon (WS–BrC) in PM2.5. The WS-BrC of TY (a sampling site on the edge of Tangyu of Mei county) has a stronger light absorption ability than CH (a rural sampling site, near the Cuihua Mountains scenic spot). The direct radiation effect of WS-BrC relative to elemental carbon (EC) is 6.67 ± 1.36% in TY and 24.13 ± 10.84% in CH in the UV range, respectively. In addition, two humic-like and one protein-like fluorophore components in WS-BrC were identified by fluorescence spectrum and parallel factor (EEMs-PARAFAC). Humification index (HIX), biological index (BIX) and fluorescence index (FI) together showed that the WS-BrC in the two sites may originate from fresh aerosol emissions. Potential source analysis of Positive Matrix Factorization (PMF) model show that the combustion process, vehicle, secondary formation and road dust are the main contributors to WS-BrC.

Key factors driving dissolved organic matter composition and bioavailability in lakes situated along the Eastern Route of the South-to-North Water Diversion Project, China

The Eastern Route of the South-to-North Water Diversion Project (SNWDP-ER) is a large scale multi-decade infrastructure project aiming to divert substantial amounts of water (≈45 billion m3 yr−1) to alleviate water shortage in comparatively arid regions of northern China. The project has ramifications for hydrological connectivity and biogeochemical cycling of dissolved organic matter (DOM) in regional lakes affected by the project. We carried out an extensive field sampling campaign along the SNWDP-ER in different hydrological seasons of 2018 and monthly observations in Lake Hongze and Lake Luoma from April 2018 to June 2021. We found the lakes connecting to the SNWDP-ER had higher mean DOC, specific UV absorbance, higher ratio of humic-like to protein-like fluorophores (Humic : Protein), and shallower spectral slope (S275–295) in the wet season compared to the wet-to-dry transition, and dry seasons. The southern lakes and Yangtze River had lower DOC concentration, bioavailable DOC (BDOC), and higher DOM aromaticity compared to the northern two downstream lakes. Ultrahigh-resolution mass spectrometry (FT-ICR MS) revealed higher relative abundance of CHO-containing and aromatic compounds in the Yangtze River and the southern three upstream lakes compared to the northern two lakes. The data from Lake Hongze and Lake Luoma, studied in different hydrological seasons, suggest that water delivery had high consistency in DOM composition and BDOC over the season. We conclude that positioning along the watercourse and seasonally variable hydrological conditions play an important role in influencing the DOM composition and bioavailability of key lakes connecting to the SNWDP-ER. Our results indicated that the water diversion project delivers water with low DOC concentration and higher aromaticity and thus is of higher quality since it has higher DOM removal potential during drinking water treatment.

A data-driven approach for understanding the structure dependence of redox activity in humic substances

Humic substances (HS) can facilitate electron transfer during biogeochemical processes due to their redox properties, but the structure-redox activity relationships are still difficult to describe and poorly understood. Herein, the linear (Partial Least Squares regressions; PLS) and nonlinear (artificial neural network; ANN) models were applied to monitor the structure dependence of HS redox activities in terms of electron accepting (EAC), electron donating (EDC) and overall electron transfer capacities (ETC) using its physicochemical features as input variables. The PLS model exhibited a moderate ability with R2 values of 0.60, 0.53 and 0.65 to evaluate EAC, EDC and ETC, respectively. The variable influence in the projection (VIP) scores of the PLS identified that the phenols, quinones and aromatic systems were particularly important for describing the redox activities of HS. Compared with the PLS model, the back-propagation ANN model achieved higher performance with R2 values of 0.81, 0.65 and 0.78 for monitoring the EAC, EDC and ETC, respectively. Sensitivity analysis of the ANN separately identified that the EAC highly depended on quinones, aromatics and protein-like fluorophores, while the EDC depended on phenols, aromatics and humic-like fluorophores (or stable free radicals). Additionally, carboxylic groups were the best indicator for evaluating both the EAC and EDC. Good model performances were obtained from the selected features via the PLS and sensitivity analysis, further confirming the accuracy of describing the structure-redox activity relationships with these analyses. This study provides a potential approach for identifying the structure-activity relationships of HS and an efficient machine-learning model for predicting HS redox activities.

Using a stable isotope tracing technique to elucidate the effect of substrate C/N ratio on the formation of different constituents of extracellular polymeric substances in an aerobic-anoxic sequencing batch reactor

Carbon to nitrogen (C/N) ratios in wastewater vary with time and sources, which greatly affect the production and composition of extracellular polymeric substances (EPS) in biological treatment processes. In this study, the EPS yield and their compositional changes were examined using stable isotopes (13C-glucose and 15NH4Cl) during the operation of aerobic-anoxic sequencing batch reactors (SBR) with different C/N ratios (C/N = 5, 10, and 15). Spectroscopic characterization revealed that humic-like aromatic compounds tended to be more enriched in loosely bound EPS (LB-EPS) at a lower C/N ratio, whereas protein-like fluorophores prevailed in tightly bound EPS (TB-EPS) (>90 %) regardless of the feed C/N ratio. Variations in isotopic enrichment revealed that the rates of substrate assimilation into TB-EPS were different between organic carbon and nitrogen, with an earlier replenishment of nitrogen regardless of C/N ratios. Yield estimation based on individual carbon and nitrogen isotopes indicated that maximum 17.8 % of organic carbon was converted into TB-EPS at a lower C/N ratio, whereas nitrogenous EPS exhibited higher yield coefficients with a wider range from 26.0 % (C/N = 5) to 44.0 % (C/N = 15). The results suggest that varying C/N ratios might exert more pronounced effects on the production of nitrogenous EPS constituents than their carbon counterparts. This suggested isotope labeling approach can be further applied to determine the mass balances among the substrate, biomass, and bound/soluble EPS within activated sludge systems.

Effects of desloratadine on activated sludge: Behaviour of EPS and sludge properties

Desloratadine (DESL), a second generation of antihistamines, is an important anti-allergic pharmaceutical used to treat allergic rhinitis, hay fever and urticaria. In this study, the overall performance, extracellular polymeric substances (EPS) production, and sludge properties were assessed in a sequencing batch reactor wastewater treatment process with activated sludge during 139 days, under the presence of DESL (1, 5, and 10 mg L−1). DESL at 10 mg L−1 impacted biomass activity decreasing the chemical oxygen demand removal (78%) and the ammonium removal (71%). The removal of DESL was of 63%. Tightly bound EPS (TB-EPS) was significantly higher (149.5 mg gMLVSS−1) at the end of operation. Peaks attributed to protein-like fluorophores clearly predominated along the experimental phases using three-dimensional excitation-emission matrix (3D-EEM) fluorescence. The peak locations and intensities in the EPS fluorescence revealed the difference in the chemical structures of the EPS caused by DESL exposure. Quantitative image analysis results clearly demonstrated the formation of large aggregates. Principal component analysis (PCA) showed a positive relationship between TB-EPS components, and large aggregates. Moreover, the results allowed to distinguish the different operational phases, emphasizing the effect of DESL on EPS and aggregates.

Measurement report: Optical properties and sources of water-soluble brown carbon in Tianjin, North China – insights from organic molecular compositions

Abstract. Brown carbon (BrC) aerosols exert vital impacts on climate change and atmospheric photochemistry due to their light absorption in the wavelength range from near-ultraviolet (UV) to visible light. However, the optical properties and formation mechanisms of ambient BrC remain poorly understood, limiting the estimation of their radiative forcing. In the present study, fine particles (PM2.5) were collected during 2016–2017 on a day/night basis over urban Tianjin, a megacity in northern China. Light absorption and fluorescence properties of water extracts of PM2.5 were investigated to obtain seasonal and diurnal patterns of atmospheric water-soluble BrC. There were obvious seasonal, but no evident diurnal, variations in the light absorption properties of BrC. In winter, BrC showed much stronger light-absorbing ability, with a mass absorption efficiency at 365 nm (MAE365) in winter (1.54±0.33 m2 gC−1) that was 1.8 times larger than MAE365 in summer (0.84±0.22 m2 gC−1). Direct radiative effects by BrC absorption relative to black carbon in the UV range were 54.3±16.9 % and 44.6±13.9 % in winter and summer, respectively. In addition, five fluorescent components in BrC, including three humic-like fluorophores and two protein-like fluorophores were identified with excitation–emission matrix fluorescence spectrometry and parallel factor (PARAFAC) analysis. The less oxygenated components contributed more to winter and nighttime samples, while more oxygenated components increased in summer and daytime samples. The higher humification index (HIX), together with lower biological index (BIX) and fluorescence index (FI), suggests that the chemical compositions of BrC were associated with a high aromaticity degree in summer and daytime due to photobleaching. Fluorescent properties indicate that wintertime BrC were predominantly affected by primary emissions and fresh secondary organic aerosol (SOA), while summer ones were more influenced by aging processes. Results of source apportionments using organic molecular compositions of the same set of aerosols reveal that fossil fuel combustion and aging processes, primary bioaerosol emission, biomass burning, and biogenic and anthropogenic SOA formation were the main sources of BrC. Biomass burning contributed much more to BrC in winter and at nighttime, while biogenic SOA contributed more in summer and during the daytime. In particular, our study highlights that primary bioaerosol emission is an important source of BrC in urban Tianjin in summer.

Effects of pyrolysis temperature on proton and cadmium binding properties onto biochar-derived dissolved organic matter: Roles of fluorophore and chromophore

Understanding the environmental behavior of biochar-derived dissolved organic matter (BDOM) is crucial for promoting the extensive utilization of biochar and meeting the carbon neutrality targets. However, limited studies focused on the binding mechanism of protons and Cd with DOM released from biochar produced at different pyrolysis temperatures. By combining excitation-emission matrix spectroscopy and parallel factor analysis, we found that the humic-like fluorophores in BDOM had higher aromaticity, molecular weight, and contents of carboxylic and phenolic groups relative to the protein-like fluorophores. Conversely, the protein-like fluorophores exhibited a stronger binding affinity for Cd than humic-like fluorophores. With the pyrolysis temperature increased from 300 °C to 500 °C, the quenching effects of Cd on the protein-like components were enhanced significantly. Their fluorescence intensities could be quenched up to 51.64%. The results of ultraviolet–visible absorbance spectroscopy and differential absorbance spectroscopy showed that the carboxylic-like and phenolic-like chromophores were involved in the protons and Cd binding process of BDOM. The binding ability of phenolic-like chromophores with Cd was reduced as a function of increasing pyrolysis temperature. These findings implied that these carboxylic and phenolic groups were mainly contained in the non-fluorescent components. Besides, protons and Cd could also induce inter-chromophore interactions in BDOM, and the interaction was proportional to the pyrolysis temperature. These results clearly demonstrated the pyrolysis temperature-dependent changes in the protons and Cd binding properties of BDOM. More importantly, the possible risk of Cd mobility caused by the protein-like components in BDOM cannot be ignored when the biochar was applied in contaminated soils. This research extends our knowledge of the application potentiality of biochar in heavy metal polluted soil.

Impact of permeate flux and gas sparging rate on membrane performance and process economics of granular anaerobic membrane bioreactors

This research investigated the impact of permeate flux and gas sparging rate on membrane permeability, dissolved and colloidal organic matter (DCOM) rejection and process economics of granular anaerobic membrane bioreactors (AnMBRs). The goal of the study was to understand how membrane fouling control strategies influence granular AnMBR economics. To this end, short- and long-term filtration tests were performed under different permeate flux and specific gas demand (SGD) conditions. The results showed that flux and SGD conditions had a direct impact on membrane fouling. At normalised fluxes (J20) of 4.4 and 8.7 L m−2 h−1 (LMH) the most favourable SGD condition was 0.5 m3 m−2 h−1, whereas at J20 of 13.0 and 16.7 LMH the most favourable SGD condition was 1.0 m3 m−2 h−1. The flux and the SGD did not have a direct impact on DCOM rejection, with values ranging between 31 and 44%. The three-dimensional excitation-emission matrix fluorescence (3DEEM) spectra showed that protein-like fluorophores were predominant in mixed liquor and permeate samples (67–79%) and were retained by the membrane (39–50%). This suggests that protein-like fluorophores could be an important foulant for these systems. The economic analysis showed that operating the membranes at moderate fluxes (J20 = 7.8 LMH) and SGD (0.5 m3 m−2 h−1) could be the most favourable alternative. Finally, a sensitivity analysis illustrated that electricity and membrane cost were the most sensitive economic parameters, which highlights the importance of reducing SGD requirements and improving membrane permeability to reduce costs of granular AnMBRs.

Repurposing an adenine riboswitch into a fluorogenic imaging and sensing tag.

Fluorogenic RNA aptamers are used to genetically encode fluorescent RNA and to construct RNA-based metabolite sensors. Unlike naturally occurring aptamers that efficiently fold and undergo metabolite-induced conformational changes, fluorogenic aptamers can exhibit poor folding, which limits their cellular fluorescence. To overcome this, we evolved a naturally occurring well-folded adenine riboswitch into a fluorogenic aptamer. We generated a library of roughly 1015 adenine aptamer-like RNAs in which the adenine-binding pocket was randomized for both size and sequence, and selected Squash, which binds and activates the fluorescence of green fluorescent protein-like fluorophores. Squash exhibits markedly improved in-cell folding and highly efficient metabolite-dependent folding when fused to a S-adenosylmethionine (SAM)-binding aptamer. A Squash-based ratiometric sensor achieved quantitative SAM measurements, revealed cell-to-cell heterogeneity in SAM levels and revealed metabolic origins of SAM. These studies show that the efficient folding of naturally occurring aptamers can be exploited to engineer well-folded cell-compatible fluorogenic aptamers and devices.

Variability of dissolved organic matter in two coastal wetlands along the Changjiang River Estuary: Responses to tidal cycles, seasons, and degradation processes

Dissolved organic matter (DOM) in the coastal tidal marsh–estuary systems are complex mixtures with different source materials that vary with hydrological regimes, seasons, and environmental conditions and are modified by removal processes including photochemical and microbial degradations. Here, monthly surveys of DOM and its optical properties (i.e., absorbance and fluorescence of DOM) covering a complete semi-diurnal tidal cycle were conducted in two coastal marshes with distinct hydrological regimes (i.e., one freshwater and one brackish marsh) in the Changjiang River Estuary (CRE). Four fluorescent components were identified by excitation–emission matrix fluoresces combined with parallel factor analysis (EEMs–PARAFAC) as two terrestrial humic-like components and two autochthonous protein-like components. Results indicated that ebbing waters draining the marshes were consistently enriched with highly absorbing, more humic and highly aromatic DOM compared to the flood tidewaters. On a seasonal basis, DOM dynamics were largely modulated by the marsh productivity and the seasonal Changjiang runoff. Protein-like fluorophores, however, demonstrated a constant, less variable pattern on both the tidal and seasonal timescales. Onsite water incubation experiments with photochemical and microbial alterations revealed that photochemistry was primarily responsible for the removal of optically active components in the marsh DOM pools whereas the impact of microbes was minor. Principal component analysis (PCA) illustrated the processes regulating the DOM dynamics at the marsh–estuarine interface and allowed a clear distinction of samples between the two marshes, in addition to the samples under the influence of episodic weather events (i.e., Super Typhoon Lekima in summer 2009 and the basin-wide severe flood event occurred in summer 2020). This study underscores the importance of world large-rivers such as Changjiang on estuarine marsh DOM dynamics and also highlights the substantial heterogeneity of the marsh DOM across a river-dominated estuary.

Elucidation of in-situ produced organic matrix effect on the solar photo/photocatalytic inactivation of E. coli

Solar photoinactivation and photocatalytic inactivation of E. coli in aqueous solution was investigated in various matrix conditions in the presence/absence of humic acid as representative of natural organic matter and inorganic components focusing on the spectroscopic evaluation of in-situ produced organic matrix. Characterization of organic matter with respect to E. coli inactivation under various experimental conditions was followed by the elucidation of specified UV–vis and fluorescence parameters as well as dissolved organic carbon content. Solar light initiated destruction of bacterial cells via lysis were visualized by the occurrence of protein-like fluorophores and fluorophores of microbial by-products as presented in excitation emission matrix fluorescence contour plots indicating a variety of changes in their shape and intensity with respect to increasing irradiation periods. Moreover, under similar experimental conditions, in the presence of humic matter these fluorophores were mainly quenched by humic-like fluorophores.E. coli inactivation kinetics obeyed first order kinetic model for both solar photoinactivation and photocatalytic inactivation conditions. Rate constants for photoinactivation of E.coli ranged from 0.184 to 0.0920 min−1, whereas photocatalytic inactivation rate constants varied from 0.248 to 0.0488 min−1. Both humic acid and water matrix components had inhibitory effects on inactivation efficiency which could be attributed to competitive reactions occurring between humic matter, bacterial cells, and bacteria derived organic material. Further evaluation of bacteria inactivation was followed by detection of organic (protein and carbohydrate) and inorganic (K+ leakage) constituents under specified conditions in correlation to the spectroscopic parameters.

Modelling of impact of presence/absence of suspended particulate organic matter from river and sea and effluent wastewater on fluorescence signal in the coastal area of Gapeau River.

Organic matter has an important role in biogeochemistry in aquatic environments. This study investigated impact of suspended particulate organic matter (SPOM) on fluorescence signal of mixtures of three water types (river water RW, sea water SW, effluent wastewater WW) using fluorescence (excitation-emission matrix, EEM) spectroscopy and parallel factor analysis (PARAFAC) and multilinear regression. Four irradiation experiments (Expt-1, Expt-2, Expt-3, and Expt-4) were conducted during different times of the year ( two in autumn, one in winter, and one in spring season). Samples were exposed to natural sunlight on laboratory rooftop in University of Toulon, France, with another set of samples kept in dark as control samples. Three component (C1, C2, C3) model was validated by split-half and Concordia from the whole EEM dataset of all irradiation experiments. No protein-like fluorophores was found. The study revealed the effect of SPOM presence/absence on fluorescence signal of DOM and on resulting parameters of multilinear regression MLR model and kinetic constant of these MLR parameters. Kinetic constant (k) for all MLR coefficients was in order of greatness as Expt-1 (SPOM of WW only in mixtures) > Expt-3 (SPOM of SW only in mixtures) > Expt-2 (SPOM of RW only in mixtures)> Expt-4 (SPOM of RW + SW + WW in mixtures) indicating that SPOM of WW is the most resistant to photodegradation. For dark control samples, only relative standard deviation RSD could be calculated from dataset. RSD values for C3 were the highest indicating its chaotic variations, and the lowest RSD values were found for both C1 and C2 for all experiments. Statistical differences has been found between control and irradiated experiments. These models developed in this study can be used to predict fluorescence signal of anthropogenic effluent DOM during its transport in river systems to coastal zone.

Characteristics of conservative and non-conservative CDOM of a tropical monsoonal estuary in relation to changing biogeochemistry

The absorbance and fluorescence characteristics of colored dissolved organic matter (CDOM) were investigated in the tropical monsoonal Godavari estuary over a year during 18 field surveys attempting to capture different river discharge scenarios ranging from nil to >5000 ms −1 and the accompanying environmental factors. Key biogeochemical constituents were also analyzed to explain the abundance and structure of CDOM and fluorescent DOM (FDOM) in terms of spatiotemporal controls. The high flow water of the monsoon season was rich in CDOM of spectral slope (S300−500) typical for river water but of lower fluorescence. The first flood water was most rich in CDOM of lowest S300−500, attributed to land washings during summer. The ratio between the UV–visible​ humic-like (A) and marine humic-like (M) fluorophores was linearly related to that of tryptophan protein-like (T) and Tyrosine protein-like (B) fluorophores and indicated linkage between terrestrial inputs and marine processes. The low and nil flow water was composed of seasonally and spatially variable CDOM but which was conservative against salinity (S) fitting to the model, aCDOM(350) (m−1) = 2.196-0.036*S. During the discharge season, pH held an inverse linear relationship to aCDOM(350): a350 (m−1) = 9.346-0.535*pH. The salinity co-varying (conservative) CDOM was linear versus silicate. A laboratory experiment showed that CDOM, FDOM and silicate were co-leached from the estuarine silts. During the non-discharge season unique, transient factors affected the CDOM e.g., (i) hot summer conditions in the upper estuary, (ii) benthic flux by bacterial metabolism from the consolidated silts after the flood season, and (iii) occasional anthropogenic input of petroleum hydrocarbons. The B:T fluorophore ratio was higher and linear versus NH4+ during no-discharge and is a potential index of the bacterial metabolism i.e., protein un-folding of the silt-held organic matter accumulated during the erstwhile flood season.

Fluorescence analysis allows to predict the oxidative capacity of humic quinones in dissolved organic matter: implication for pollutant degradation

Dissolved organic matter (DOM) controls the degradation and sequestration of aquatic pollutants and, in turn, water quality. In particular, pollutant degradation is performed by oxidant species that are generated by exposure of DOM to solar light, yet, since DOM is a very complex mixture of poorly known substances, the relationships between potential oxidant precursors in DOM and their oxydative capacity is poorly known. Here, we hypothesized that production of oxidant species could be predicted using fluorescence analysis. We analysed water samples from an alluvial plain by fluorescence spectroscopy; the three-dimensional spectra were then decomposed into seven individual components using a multi-way algorithm. Components include a protein-like fluorophore, e.g. tryptophan-like and tyrosine-like, three humic fluorophores, 2-naphthoxyacetic acid, and a by-product. We compared component levels with the ability of water samples to generate reactive species under solar light. The results show a strong correlation between reactive species production and the intensity of two humic-like fluorophores assigned to reduced quinones. Monitoring these fluorophores should thus allow to predict the ability of DOM degradation of pollutants in surface waters.