Yuanrong Zhu

Characterization of Organic Phosphorus in Lake Sediments by Sequential Fractionation and Enzymatic Hydrolysis

The role of sediment-bound organic phosphorus (Po) on lake eutrophication was studied using sequential extraction and enzymatic hydrolysis by collecting sediments from Dianchi Lake, China. Bioavailable Po species including labile monoester P, diester P, and phytate-like P were identified in the sequential extractions by H2O, NaHCO3, and NaOH. For the H2O-Po, 36.7% (average) was labile monoester P, 14.8% was diester P, and 69.9% was phytate-like P. In NaHCO3-Po, 19.9% was labile monoester P, 17.5% was diester P, and 58.8% was phytate-like P. For NaOH-Po, 25.6% was labile monoester P, 7.9% was diester P, and 35.9% was phytate-like P. Labile monoester P was active to support growth of algae to form blooms. Diester P mainly distributed in labile H2O and NaHCO3 fractions was readily available to cyanobacteria. Phytate-like P represents a major portion of the Po in the NaOH fractions, also in the more labile H2O and NaHCO3 fractions. Based on results of sequential extraction of Po and enzymatic hydrolysis, lability and bioavailability was in decreasing order as follows: H2O-Po > NaHCO3-Po > NaOH-Po, and bioavailable Po accounted for only 12.1-27.2% of total Po in sediments. These results suggest that the biogeochemical cycle of bioavailable Po might play an important role in maintaining the eutrophic status of lakes.

Characteristics and degradation of carbon and phosphorus from aquatic macrophytes in lakes: Insights from solid-state (13)C NMR and solution (31)P NMR spectroscopy.

Water extractable organic matter (WEOM) derived from macrophytes plays an important role in biogeochemical cycling of nutrients, including carbon (C), nitrogen (N) and phosphorus (P) in lakes. However, reports of their composition and degradation in natural waters are scarce. Therefore, compositions and degradation of WEOM derived from six aquatic macrophytes species of Tai Lake, China, were investigated by use of solid-state (13)C NMR and solution (31)P NMR spectroscopy. Carbohydrates were the predominant constituents of WEOM fractions, followed by carboxylic acid. Orthophosphate (ortho-P) was the dominant form of P (78.7% of total dissolved P) in the water extracts, followed by monoester P (mono-P) (20.6%) and little diester P (0.65%). The proportion of mono-P in total P species increased with the percentage of O-alkyl and O-C-O increasing in the WEOM, which is likely due to degradation and dissolution of biological membranes and RNA from aquatic plants. Whereas the proportion of mono-P decreased with alkyl-C, NCH/OCH3 and COO/N-C=O increasing, which may be owing to the insoluble compounds including C functional groups of alkyl-C, NCH/OCH3 and COO/N-C=O, such as aliphatic biopolymers, lignin and peptides. Based on the results of this study and information in the literature about water column and sediment, we propose that WEOM, dominated by polysaccharides, are the most labile and bioavailable component in debris of macrophytes. Additionally, these WEOMs would also be a potential source for bioavailable organic P (e.g., RNA, DNA and phytate) for lakes.

Forms and Lability of Phosphorus in Algae and Aquatic Macrophytes Characterized by Solution 31P NMR Coupled with Enzymatic Hydrolysis

Solution Phosphorus-31 nuclear magnetic resonance (31P NMR) spectroscopy coupled with enzymatic hydrolysis (EH) with commercially available phosphatases was used to characterize phosphorus (P) compounds in extracts of the dominant aquatic macrophytes and algae in a eutrophic lake. Total extractable organic P (Po) concentrations ranged from 504 to 1643 mg kg−1 and 2318 to 8395 mg kg−1 for aquatic macrophytes and algae, respectively. Using 31P NMR spectroscopy, 11 Po species were detected in the mono- and diester region. Additionally, orthophosphate, pyrophosphate and phosphonates were also detected. Using EH, phytate-like P was identified as the prevalent class of enzyme-labile Po, followed by labile monoester- and diester-P. Comparison of the NMR and EH data indicated that the distribution pattern of major P forms in the samples determined by the two methods was similar (r = 0.712, p < 0.05). Additional 31P NMR spectroscopic analysis of extracts following EH showed significant decreases in the monoester and pyrophosphate regions, with a corresponding increase in the orthophosphate signal, as compared to unhydrolyzed extracts. Based on these quantity and hydrolysis data, we proposed that recycling of Po in vegetative biomass residues is an important mechanism for long-term self-regulation of available P for algal blooming in eutrophic lakes.

Characterization of plant-derived carbon and phosphorus in lakes by sequential fractionation and NMR spectroscopy.

Although debris from aquatic macrophytes is one of the most important endogenous sources of organic matter (OM) and nutrients in lakes, its biogeochemical cycling and contribution to internal load of nutrients in eutrophic lakes are still poorly understood. In this study, sequential fractionation by H2O, 0.1M NaOH and 1.0M HCl, combined with (13)C and (31)P NMR spectroscopy, was developed and used to characterize organic carbon (C) and phosphorus (P) in six aquatic plants collected from Tai Lake (Ch: Taihu), China. Organic matter, determined by total organic carbon (TOC), was unequally distributed in H2O (21.2%), NaOH (29.9%), HCl (3.5%) and residual (45.3%) fractions. For P in debris of aquatic plants, 53.3% was extracted by H2O, 31.9% by NaOH, and 11% by HCl, with 3.8% in residual fractions. Predominant OM components extracted by H2O and NaOH were carbohydrates, proteins and aliphatic acids. Inorganic P (Pi) was the primary form of P in H2O fractions, whereas organic P (Po) was the primary form of P in NaOH fractions. The subsequent HCl fractions extracted fewer species of C and P. Some non-extractable carbohydrates, aromatics and metal phytate compounds remained in residual fractions. Based on sequential extraction and NMR analysis, it was proposed that those forms of C (54.7% of TOC) and P (96.2% of TP) in H2O, NaOH and HCl fractions are potentially released to overlying water as labile components, while those in residues are stable and likely preserved in sediments of lakes. These results will be helpful in understanding internal loading of nutrients from debris of aquatic macrophytes and their recycling in lakes.

Cation-induced coagulation of aquatic plant-derived dissolved organic matter: Investigation by EEM-PARAFAC and FT-IR spectroscopy

Complexation and coagulation of plant-derived dissolved organic matter (DOM) by metal cations are important biogeochemical processes of organic matter in aquatic systems. Thus, coagulation and fractionation of DOM derived from aquatic plants by Ca(II), Al(III), and Fe(III) ions were investigated. Metal ion-induced removal of DOM was determined by analyzing dissolved organic carbon in supernatants after addition of these metal cations individually. After additions of metal ions, both dissolved and coagulated organic fractions were characterized by use of fluorescence excitation emission matrix-parallel factor (EEM-PARAFAC) analysis and Fourier transform infrared (FT-IR) spectroscopy. Addition of Ca(II), Fe(III) or Al(III) resulted in net removal of aquatic plant-derived DOM. Efficiencies of removal of DOM by Fe(III) or Al(III) were greater than that by Ca(II). However, capacities to remove plant-derived DOM by the three metals were less than which had been previously reported for humic materials. Molecular and structural features of plant-derived DOM fractions in associations with metal cations were characterized by changes in fluorescent components and infrared absorption peaks. Both aromatic and carboxylic-like organic matters could be removed by Ca(II), Al(III) or Fe(III) ions. Whereas organic matters containing amides were preferentially removed by Ca(II), and phenolic materials were selectively removed by Fe(III) or Al(III). These observations indicated that plant-derived DOM might have a long-lasting effect on water quality and organisms due to its poor coagulation with metal cations in aquatic ecosystems. Plant-derived DOM is of different character than natural organic matter and it is not advisable to attempt removal through addition of metal salts during treatment of sewage.

Using solid (13)C NMR coupled with solution (31)P NMR spectroscopy to investigate molecular species and lability of organic carbon and phosphorus from aquatic plants in Tai Lake, China.

Forms and labilities of plant-derived organic matters (OMs) including carbon (C) and phosphorus (P) were fundamental for understanding their release, degradation and environmental behaviour in lake ecosystems. Thus, solid 13C and solution 31P nuclear magnetic resonance (NMR) spectroscopy were used to characterize biomass of six aquatic plants in Tai Lake, China. The results showed that carbohydrates (61.2% of the total C) were predominant C functional group in the solid 13C NMR spectra of plant biomass, which may indicate high lability and bioavailability of aquatic plants-derived organic matter in lakes. There was 72.6–103.7% of the total P in aquatic plant biomass extracted by NaOH–EDTA extracts. Solution 31P NMR analysis of these NaOH–EDTA extracts further identified several molecular species of P including orthophosphate (50.1%), orthophosphate monoesters (46.8%), DNA (1.6%) and pyrophosphate (1.4%). Orthophosphate monoesters included β-glycerophosphate (17.7%), hydrolysis products of RNA (11.7%), α-glycerophosphate (9.2%) and other unknown monoesters (2.1%). Additionally, phytate, the major form of organic P in many lake sediments, was detected in floating plant water poppy. These inorganic P (e.g. orthophosphate and pyrophosphate) and organic P (e.g. diester and its degradation products) identified in plant biomass were all labile and bioavailable P, which would play an important role in recycling of P in lakes. These results increased knowledge of chemical composition and bioavailability of OMs derived from aquatic plants in lakes.

Molecular characterization of macrophyte-derived dissolved organic matters and their implications for lakes

Chemical properties of whole organic matter (OM) and its dissolved organic matter (DOM) extracted from three types of dominant macrophytes in Lake Dianchi were comparatively characterized using elemental analysis, UV, 3D-EEM and 13C NMR spectroscopy and their implications for lakes were discussed. Ratios of C/N and C/P were least in the floating water hyacinth and submerged sago pondweed, while total dissolved nitrogen (TDN) and phosphorus (TDP), dissolved organic nitrogen (DON) and phosphorus (DOP) were greatest in those species. In emergent species, C/N, C/P, DON and DON/TDN were less in leaves than in their corresponding shoots. The specific UV absorbance at 254nm (SUVA254) and 280nm (SUVA280) of extracts were in the range of 0.50-1.96L/mgC·m and 0.40-1.48L/mgC·m. Both SUVA values were greater in leaves than those in shoots. 3D-EEM spectra showed only a single fulvic-like fluorescence in leaves of emergent macrophytes. In contrast, protein-like peak were observed in spectra of floating and submerged species, as well as the shoot DOM of emergent species. Solid-state 13C NMR demonstrated that leaves had greater percentage of recalcitrant alkyl C and aromatic C, while shoots were rich in labile carbohydrates. The overall characterization works suggested that macrophyte-derived DOM has less aromatic constituents than do DOM in natural waters and soil leachates. Also OM and DOM derived from shoots had greater contents of protein-like and carbohydrate materials, while leaves were rich in aromatics. Floating and submerged plants possessed potential to not only accumulate excess N and P, but also for returning them to the lake. Shoots of riparian and emergent species were also an important source of nutrients. Thus, macrophyte biomass should be a great concern in nutrient regulation in Lake Dianchi.

Simulated bioavailability of phosphorus from aquatic macrophytes and phytoplankton by aqueous suspension and incubation with alkaline phosphatase

Bioavailability of phosphorus (P) in biomass of aquatic macrophytes and phytoplankton and its possible relationship with eutrophication were explored by evaluation of forms and quantities of P in aqueous extracts of dried macrophytes. Specifically, effects of hydrolysis of organically-bound P by the enzyme alkaline phosphatase were studied by use of solution 31P-nuclear magnetic resonance (NMR) spectroscopy. Laboratory suspensions and incubations with enzymes were used to simulate natural releases of P from plant debris. Three aquatic macrophytes and three phytoplankters were collected from Tai Lake, China, for use in this simulation study. The trend of hydrolysis of organic P (Po) by alkaline phosphatase was similar for aquatic macrophytes and phytoplankton. Most monoester P (15.3% of total dissolved P) and pyrophosphate (1.8%) and polyphosphate (0.4%) and DNA (3.2%) were transformed into orthophosphate (14.3%). The major forms of monoester P were glycerophosphate (8.8%), nucleotide (2.5%), phytate (0.4%) and other monoesters P (3.6%). Proportions of Po including condensed P hydrolyzed in phytoplankton and aquatic macrophytes were different, with the percentage of 22.6% and 6.0%, respectively. Proportion of Po hydrolyzed in debris from phytoplankton was approximately four times greater than that of Po from aquatic macrophytes, and could be approximately twenty-five times greater than that of Po in sediments. Thus, release and hydrolysis of Po, derived from phytoplankton debris would be an important and fast way to provide bioavailable P to support cyanobacterial blooming in eutrophic lakes.

Tissue Residue Guideline for ∑DDT for Protection of Aquatic Birds in China

ABSTRACT DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) is a chlorinated hydrocarbon insecticide that has been used worldwide. While the use of DDT has been phased out in many countries, it is still produced in some parts of the world for use to control vectors of malaria. DDE (1,1,-dichloro-2,2-bis(p-chlorophenyl)ethylene) and DDD (1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) are primary metabolites of DDT and have similar chemical and physical properties. DDT and its metabolites (DDE and DDD) are collectively referred to as ∑DDT. The lipophilic nature and persistence of the ∑DDT result in biomagnification in wildlife that feed at higher trophic levels in the food chain. Wildlife in aquatic ecosystems depend on aquatic biota as their primary source of food, which provide the main route of exposure to ∑DDT. Studies about effects of ∑DDT on birds were reviewed. The tissue residue guidelines for DDT (TRGs) for protection of birds in China were derived using species sensitivity distribution (SSD) and toxicity percentile rank method (TPRM) based on the available toxicity data. Risks of ∑DDT to birds were assessed by comparing the TRGs and ∑DDT concentrations in fishes from China. The tissue residue guideline for protection of birds in China is recommended to be 12.0 ng ∑DDT/g food.

Bioavailability and preservation of organic phosphorus in lake sediments: Insights from enzymatic hydrolysis and 31P nuclear magnetic resonance

Bioavailability and preservation of organic P (Po) in the sediment profiles (DC-1 and DC-2) from Lake Dianchi, a eutrophic lake in China, were investigated by a combination of enzymatic hydrolysis and solution 31P nuclear magnetic resonance (NMR) spectroscopy. Results showed that large of Po could be extracted by NaOH-EDTA (NaOH-EDTA Po), with little Po in residues after extraction with NaOH-EDTA. Bioavailability and preservation of NaOH-EDTA Po provide key information for biogeochemical cycling of Po in sediments. The details of P species and their bioavailability in NaOH-EDTA Po showed that 54.8-70.4% in DC-1 and 54.6-100% in DC-2, measured by 31P NMR, could be hydrolyzed by the phosphatase. Whereas, some proportion of NaOH-EDTA Po could not be hydrolyzed by the phosphatase, and decreased with sediment depth. Interaction between Po and other organic matter (e.g., humic acids) is likely an important factor for preservation of these Po in the sediment profiles. Simulation experiments of hydrolysis of model Po compounds adsorbed by minerals, such as goethite and montmorillonite, further indicated that adsorption to minerals protected some Po, especially phytate-like P, from enzymatic hydrolysis, thus preserving these forms of Po in sediments. Interactions of Po with organic matter and minerals in the sediments are two important factors determining biogeochemical cycling of Po in lakes. Intervention to break the cycle of FeP and bioavailable Po (e.g., labile monoester P) in the history of eutrophication is important way to control algal blooming.