Preparation,characterization and pharmacokinetics of fluorescence labeled propylene glycol alginate sodium sulfate

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate(PSS), in rat plasma. Fluorescein isothiocyanate(FITC) was selected to label PSS, and 1, 6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS(F-PSS) through a reductive amination reaction. F-PSS was identified by UV-Vis, FT-IR and 1H-NMR spectrum. The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney(MDCK) cells. The results indicated that the labeling efficiency of F-PSS was 0.522% ± 0.0248% and the absolute bioavailability was 8.39%. F-PSS was stable in MDCK cells without obvious cytotoxicity. The method was sensitive and reliable; it showed a good linearity, precision, recovery and stability. The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples.     

Preparation,Characterization and Pharmacokinetics of Fluorescence Labeled Propylene Glycol Alginate Sodium Sulfate

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate （PSS）,in rat plasma.Fluorescein isothiocyanate （FITC） was selected to label PSS,and 1,6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS （F-PSS） through a reductive amination reaction.F-PSS was identified by UV-Vis,FT-IR and ^1H-NMR spectrum.The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney （MDCK） cells.The results indicated that the labeling efficiency of F-PSS was 0.522%±0.0248% and the absolute bioavailability was 8.39%.F-PSS was stable in MDCK cells without obvious cytotoxicity.The method was sensitive and reliable; it showed a good linearity,precision,recovery and stability.The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples.     

Effects of Nitrogen and Phosphorus on the Growth of Levanderina fissa: How It Blooms in Pearl River Estuary

Effects of nitrogen(N) and phosphorus(P) from different sources and at different concentrations on the growth of Levanderina fissa(= Gyrodinium instriatum) were studied in laboratory conditions. The findings might explain the recurrent blooms of this species in Pearl River Estuary, China. Results showed that nutrient limitation significantly inhibited the growth of L. fissa. The values of specific growth rate(μmax) and half-saturation nutrient concentration(KS) were 0.37 divisions/d and 8.49 μmol L~(-1) for N, and 0.39 divisions/d and 1.99 μmol L~(-1) for P, respectively. Based on KS values, dissolved inorganic N level in PRE was sufficient to support the high proliferation of L. fissa, while dissolved inorganic P concentration was far lower than the minimum requirement for its effective growth. L. fissa was not able to utilize dissolved organic N(DON) compounds such as urea, amino acids, and uric acid. However, it grew well by using a wide variety of dissolved organic P(DOP) sources like nucleotides, glycerophosphate, and 4-nitrophenylphosphate. The results from this study suggested that the ability in DOP utilization of L. fissa offers this species a competitive advantage in phytoplankton communities. The high level and continuous supply of DIN, enrichment of DOP, together with warm climate and low salinity in the Pearl River Estuary provided a suitable nutrient niche for the growth of L. fissa, and resulted in the recurrent blooms in the estuary.     

α-Glucosidase Inhibitory Activities of Lutein and Zeaxanthin Purified from Green Alga Chlorella ellipsoidea

α-Glucosidase inhibitors are used therapeutically to treat type-2 diabetes mellitus. Through a bioassay-guided fractionation technique, three carotenoids,(all-E)-lutein,(all-E)-zeaxanthin and(9-Z)-zeaxanthin, were purified from the green alga Chlorella ellipsoidea, in which(all-E)-lutein and(9-Z)-zeaxanthin had potent α-glucosidase inhibitory activity. IC_(50) values of(all-E)-lutein and(9-Z)-zeaxanthin were 70 and 53.5 μmol L~(-1) against Saccharomyces cerevisiae α-glucosidase, respectively, with non-competitive inhibition. In addition, IC_(50) values of(9-Z)-zeaxanthin against Bacillus stearothermophilus and rat-intestinal α-glucosidase were 805.1 and 671.2 μmol L~(-1), respectively. The K_i values of(all-E)-lutein and(9-Z)-zeaxanthin against S. cerevisiae α-glucosidase were 78.1 and 16.5 μmol L~(-1), respectively. Therefore, C. ellipsoidea carotenoids might be utilized as a novel candidate to prevent type-2 diabetes mellitus related disorders in food and medical industry.     

Purification and Characterization of an Alkaline Protease from Micrococcus sp.Isolated from the South China Sea

Protease is wildly used in various fields,such as food,medicine,washing,leather,cosmetics and other industrial fields.In this study,an alkaline protease secreted by Micrococcus NH54PC02 isolated from the South China Sea was purified and characterized.The growth curve and enzyme activity curve indicated that the cell reached a maximum concentration at the 30 th hour and the enzyme activity reached the maximum value at the 36 th hour.The protease was purified with 3 steps involving ammonium sulfate precipitation,ion-exchange chromatography and hydrophobic chromatography with 8.22-fold increase in specific activity and 23.68% increase in the recovery.The molecular mass of the protease was estimated to be 25 k Da by SDS-PAGE analysis.The optimum temperature and p H for the protease activity were 50℃ and pH 10.0,respectively.The protease showed a strong stability in a wide range of pH values ranging from 6.0–11.0,and maintained 90% enzyme activity in strong alkaline environment with p H 11.0.Inhibitor trials indicated that the protease might be serine protease.But it also possessed the characteristic of metalloprotease as it could be strongly inhibited by EDTA and strongly stimulated by Mn~(2+).Evaluation of matrix-assisted laser desorption ionization/time-of-flight MS(MALDI-TOF-TOF/MS) showed that the protease might belong to the peptidase S8 family.     

Toxicity of five phenolic compounds to brine shrimp Artemia sinica (Crustacea: Artemiidae)

The acute toxicity of five phenolic compounds each to 15 d old Artemia sinica was determined in this study. The brine shrimp A. sinica was hatched from the encysted dry eggs(Bohai Bay Brand) produced by Dongying Ocean Artemia Co., Ltd., China at 27℃± 1℃ in pre-filtered(through pores of 0.45 μm in diameter) and autoclaved seawater(salinity 31, pH 7.5–8.0) in a cilindroconical glass beaker(2000 mL in volume) under continuous illumination(provided by a side set 20 W fluorescent lamp) with slight aeration. Ten Artemia individuals from the same batch of the hatched were cultured in 10 mL toxicant solution prepared with seawater(salinity 31, pH 7.5–8.0) at room temperature(about 20℃) to determine 24 h, 48 h and 72 h medium lethal concentration(LC50) of 5 phenolic compounds each. It was found that the toxicity of n-heptylphenol was the highest followed by nonylphenol, t-butylphenol, 2,4-dichlorophenol and bisphenol A in order. The LC50 values of the 5 compounds were calculated with regression analysis. The real concentration(in μg- L1) of 5 phenolic compounds each in toxicant solutions was measured with GC/MS analysis. Significant loss of phenolic compounds caused by either adsorption or desorption was not found. The significant difference of LC50 values was found among the five compounds 3 exposure times each. The range between the highest no-observed-effect concentration(NOEC) and 100% death causing concentration of five phenolic compounds each was determined. The toxicity in term of 24 h LC50 value of n-HP was 9.10 times higher than that of BPA, 1.71 times higher than t-BP, 1.53 times higher than 2,4-DCP and 1.36 times higher than NP, respectively.     

Toxicity of Five Phenofic Compounds to Brine ShrimpArtemia sinica （Crustacea： Artemiidae）

The acute toxicity of five phenolic compounds each to 15 d old Artemia sinica was determined in this study. The brineshrimp A. sinica was hatched from the encysted dry eggs （Bohai Bay Brand） produced by Dongying Ocean Artemia Co., Ltd., China at27 °C ± 1 °C in pre-filtered （through pores of 0.45 μm in diameter） and autoclaved seawater （salinity 31, pH 7.5-8.0） in a cilindroconicalglass beaker （2000mL in volume） under continuous illumination （provided by a side set 20W fluorescent lamp） with slight aeration.Ten A rtemia individuals from the same batch of the hatched were cultured in 10 mL toxicant solution prepared with seawater （salinity31, pH 7.5-8.0） at room temperature （about 20°C） to determine 24h, 48h and 72h medium lethal concentration （LCs0） of 5 phenoliccompounds each. It was found that the toxicity of n-heptylphenol was the highest followed by nonylphenol, t-butylphenol,2,4-dichlorophenol and bisphenol A in order. The LC50 values of the 5 compounds were calculated with regression analysis. The realconcentration （in μg L-1） of 5 phenolic compounds each in toxicant solutions was measured with GC/MS analysis. Significant loss ofphenolic compounds caused by either adsorption or desorption was not found. The significant difference of LCs0 values was foundamong the five compounds 3 exposure times each. The range between the highest no-observed-effect concentration （NOEC） and 100%death causing concentration of five phenolic compounds each was determined. The toxicity in term of 24 h LC50 value of n-HP was 9.10times higher than that of BPA, 1.71 times higher than t-BP, 1.53 times higher than 2,4-DCP and 1.36 times higher than NP, respectively.     

Effects of Nitrogen Sources and Concentrations on the Growth of Different Phytoplankton Taxa

Effects of different nitrogen(N) compounds and concentrations on the growth of the three different phytoplankton taxa, Skeletonema costatum(Bacillariophyceae), Prorocentrum micans(Dinophyceae), and Chattonella marina(Raphidophyceae), were investigated. The Monod equation was applied to examine effects of N concentrations on the growth of algal cells. Results showed that nitrate(NO_3~-N) and urea served as good N sources for the three phytoplankton taxa. S. costatum grew well on all of the seven N sources. C. marina can effectively use the two amino acids, glycine(Gly) and serine(Ser), however cannot utilize alanine(Ala), threonine(Thr), and asparaginic acid(Asp). P. micans cannot grow in five amino acid substrates. All of the three phytoplankton taxa grew well under different proportions of urea-N, and C. marina grew significantly better in medium with both NO_3~-and urea-N. The values of maximum growth rate(μmax) and half-saturation nutrient concentration(KS) for NO_3~-N were 0.71 divisions d~(-1) and 53.55 μmol L~(-1) for S. costatum, 0.67 divisions d~(-1) and 23.31 μmol L~(-1) for P. micans, and 0.23 divisions d~(-1) and 17.57 μmol L~(-1) for C. marina, respectively. The results suggested that S. costatum had a high N demand for growth, and was capable of using wide ranges of N compounds. The strategy of N utilization for S. costatum may make this species an advantage in N-enriched sea areas especially the dissolved organic nitrogen(DON) rich coastal waters, which might be the reason why S. costatum widely distributes in the cosmopolitan coastal and estuarine sea areas.     

Antialgal and Antilarval Activities of Bioactive Compounds Extracted from the Marine Dinoflagellate Amphidinium carterae

With the global ban on the application of organotin-based marine coatings by the International Maritime Organization, the development of environmentally friendly, low-toxic and nontoxic antifouling compounds for marine industries has become an urgent need. Marine microorganisms have been considered as a potential source of natural antifoulants. In this study, the antifouling potential of marine dinoflagellate Amphidinium carterae, the toxic and red-tide microalgae, was investigated. We performed a series of operations to extract the bioactive substances from Amphidinium carterae and tested their antialgal and antilarval activities. The crude extract of Amphidinium carterae showed significant antialgal activity and the EC50 value against Skeletonema costatum was 55.4 μg m L~(-1). After purification, the isolated bioactive substances(the organic extract C) exhibited much higher antialgal and antilarval activities with EC50 of 12.9 μg m L~(-1) against Skeletonema costatum and LC50 of 15.1 μg m L~(-1) against Amphibalanus amphitrite larvae. Subsequently, IR, Q-TOFMS, and GC-MS were utilized for the structural elucidation of the bioactive compounds, and a series of unsaturated and saturated 16- to 22-carbon fatty acids were detected. The data suggested the bioactive compounds isolated from Amphidinium carterae exhibited a significant inhibiting effect against the diatom Skeletonema costatum and Amphibalanus amphitrite larvae, and could be substitutes for persistent, toxic antifouling compounds.     

Exploration of Antifouling Potential of the Brown Algae Laminaria ‘Sanhai’

Seaweeds are one of the largest producers of biomass in the marine environment.It has been well known that marine algae,especially brown algae was a rich source of biogenic compounds with antifouling potential that could be ideal alternatives of tributyltin(TBT).In this paper,antifouling potential of the brown algae Laminaria ‘sanhai' was explored.Firstly,the dried alga was extracted and the antialgal and antilarval activities were investigated.The EC_(50) and LC_(50) values of crude extract of Laminaria ‘sanhai' against diatom(Skeletonema costatum) and barnacle larval(Chthamalus challengeri) were 8.9 μg mL~(-1) and 12.0 μg mL~(-1) respectively.Then,guided by bioassay,the bioactive substances were isolated by liquid-liquid extraction.The antialgal and antilarval activities of isolated fraction were improved with the EC_(50) value of 7.4 μg mL~(-1) against S.costatum and LC_(50) value of 9.7 μg m L~(-1) against C.challengeri larvae.Identification by IR,Q-TOFMS and GC-MS of the isolated bioactive substances revealed the abundance of fatty acids.These fatty acids,most with 16,18 or 20 carbon atoms,contained myristic,hexadecanoic,oleic,linolenic,arachidonic and eicosapentaenoic acids.The results indicated that both the crude extract and the isolated bioactive substances had high antialgal and antilarval activities with no highlighted cytotoxicity which made the brown algae Laminaria ‘sanhai' a promising source of the environmentally friendly antifoulants.     

Evaluation ofAntitumor, Immunomodulatory and Free Radical Scavenging Effects of A New Herbal Prescription Seaweed Complex Preparation

Seaweed Complex Preparation （SCP） is a clinical traditional Chinese medicine preparation which is composed of seven traditional Chinese herbs, and it has been used for treatment of lung cancer, liver cancer and digestive cancer. However, little infor- mation is available about the pharmacodynamic basis. The antitumor, immunomodulatory and free radical scavenging effects of SCP were evaluated in this study. Transplanted tumor in vivo method was used to determine the antitumor effect. The effects on spleno- cyte proliferation and phagocytosis of macrophages in tumor-bearing mice were measured by the MTT method and the phagocytizing cock red blood cell （CRBC） method respectively. The scavenging activities of SCP on DPPH and hydroxyl radicals in vitro were investigated. It was found that the medium-dose and high-dose of SCP could significantly inhibit the growth of transplanted hepatic tumor of murine hepatocarcinoma cell line H22, and promote proliferation of splenocytes and phagocytosis of macrophages. SCP possessed noticeable scavenging activities on DPPH and hydroxyl radicals. The antiturnor effects of SCP might be achieved by im- proving immune system and scavenging free radicals, which is in accordance with the viewpoint of traditional Chinese medicine in promoting the body resistance and eliminating pathogenic factors for cancer treatment.     

Purification and Characterization of Angiotensin I Converting Enzyme Inhibition Peptides from Sandworm Sipunculus nudus

Three angiotensin I converting enzyme(ACE) inhibition peptides were isolated from sandworm Sipunculus nudus protein hydrolysate prepared using protamex. Consecutive purification methods, including size exclusion chromatography and reverse-phase high performance liquid chromatography(RP-HPLC), were used to isolate the ACE inhibition peptides. The amino acid sequences of the peptides were identified as Ile-Asn-Asp, Val-Glu-Pro-Gly and Leu-Ala-Asp-Glu-Phe. The IC_(50) values of the purified peptides for ACE inhibition activity were 34.72 μmol L~(-1), 20.55 μmol L~(-1) and 22.77 μmol L~(-1), respectively. These results suggested that S. nudus proteins contain specific peptides that can be released by enzymatic hydrolysis. This study may provide an experimental basis for further systematic research, rational development and clinical utilization of sandworm resources.     

An evaluation on using soil aggregate stability as the indicator of interrill erodibility

Aggregate stability is a very important predictor of soil structure and strength,which influences soil erodibility.Several aggregate stability indices were selected for estimating interrill erodibility of four soil types with contrasting properties from temperate and subtropical regions of China.This study was conducted to investigate how closely the soil interrill erodibility factor in the Water Erosion Prediction Project(WEPP) model relates to soil aggregate stability.The mass fractal dimension(FD),geometric mean diameter(GMD),mean weight diameter(MWD),and aggregate stability index(ASI) of soil aggregates were calculated.A rainfall simulator with a drainable flume(3.0 m long × 1.0 m wide × 0.5 m deep) was used at four slope gradients(5°,10°,15° and 20°),and four rainfall intensities(0.6,1.1,1.7 and 2.5 mm/min).Results indicated that the interrill erodibility(Ki) values were significantly correlated to the indices of ASI,MWD,GMD,and FD computed from the aggregate wet-sieve data.The Ki had a strong positive correlation with FD,as well as a strong negative correlation with ASI,GMD,and MWD.Soils with a higher aggregate stability and lower fractal dimension have smaller Ki values.Stable soils were characterized by a high percentage of large aggregates and the erodible soils by a high percentage of smaller aggregates.The correlation coefficients of Ki with ASI and GMD were greater than those with FDand MWD,implying that both the ASI and GMD may be better alternative parameters for empirically predicting the soil Ki factor.ASI and GMD are more reasonable in interrill soil erodibility estimation,compared with Ki calculation in original WEPP model equation.Results demonstrate the validation of soil aggregation characterization as an appropriate indicator of soil susceptibility to erosion in contrasting soil types in China.     

Antifouling potential of bacteria isolated from a marine biofilm

Marine microorganisms are a new source of natural antifouling compounds. In this study, two bacterial strains, Kytococcus sedentarius QDG-B506 and Bacillus cereus QDG-B509, were isolated from a marine biofilm and identified. The bacteria fermentation broth could exert inhibitory effects on the growth of Skeletonema costatum and barnacle larvae. A procedure was employed to extract and identify the antifouling compounds. Firstly, a toxicity test was conducted by graduated pH and liquid-liquid extraction to determine the optimal extraction conditions. The best extraction conditions were found to be pH 2 and 100% petroleum ether. The EC50 value of the crude extract of K. sedentarius against the test microalgae was 236.7 ± 14.08 μg mL-1, and that of B. cereus was 290.6 ± 27.11 μg mL-1. Secondly, HLB SPE columns were used to purify the two crude extracts. After purification, the antifouling activities of the two extracts significantly increased: the EC50 of the K. sedentarius extract against the test microalgae was 86.4 ± 3.71 μg mL-1, and that of B. cereus was 92.6 ± 1.47 μg mL-1. These results suggest that the metabolites produced by the two bacterial strains are with high antifouling activities and they should be fatty acid compounds. Lastly, GC-MS was used for the structural elucidation of the compounds. The results show that the antifouling compounds produced by the two bacterial strains are myristic, palmitic and octadecanoic acids.     

Phenylquinolinones with antitumor activity from the Indian Ocean-derived fungus Aspergillus versicolor Y31-2

Two phenylquinolinones,including one new compound(1) and a previously isolated compound(2),were isolated from the ethyl acetate extracts of the fungus Aspergillus versicolor Y31-2,which was obtained from seawater samples collected from the Indian Ocean.The structures of these compounds were established by spectroscopic analyses.4-(3-Hydroxyphenyl)-3-methoxyquinolin-2(1H)-one(1) exhibited moderate cytotoxicity against MCF-7(human breast carcinoma cell line) and SMMC-7721(human liver cancer cell line) cells with IC_(50) values of 16.6 and 18.2 μmol/L,respectively.To the best of our knowledge,this study represents the first reported account of the isolation of compounds 1 and 2 as the secondary metabolites of the seawater derived fungus Aspergillus versicolor from the Indian Ocean.     

Debris flows risk analysis and direct loss estimation: the case study of Valtellina di Tirano,Italy

andslide risk analysis is one of the primary studies providing essential instructions to the subsequent risk management process. The quantification of tangible and intangible potential losses is a critical step because it provides essential data upon which judgments can be made and policy can be formulated. This study aims at quantifying direct economic losses from debris flows at a medium scale in the study area in Italian Central Alps. Available hazard maps were the main inputs of this study. These maps were overlaid with information concerning elements at risk and their economic value. Then, a combination of both market and construction values was used to obtain estimates of future economic losses. As a result, two direct economic risk maps were prepared together with risk curves, useful to summarize expected monetary damage against the respective hazard probability. Afterwards, a qualitative risk map derived using a risk matrix officially provided by the set of laws issued by the regional government, was prepared. The results delimit areas of high economic as well as strategic importance which might be affected by debris flows in the future. Aside from limitations and inaccuracies inherently included in risk analysis process, identification of high risk areas allows local authorities to focus their attention on the “hot-spots”, where important consequences may arise and local （large） scale analysis needs to be performed with more precise cost-effectiveness ratio. The risk maps can be also used by the local authorities to increase population’s adaptive capacity in the disaster prevention process.     

Determination of Steroid Hormone Residues in Fish Tissues Using Gel Permeation Chromatography and Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

A highly sensitive method was developed for the simultaneous determination of 8 steroid hormones in high-fat fish tissues using ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The 8 steroid hormones were extracted from the tissues with diethyl ether.Differing from other common purification methods,the extract solutions were cleaned by gel permeation chromatography(GPC) using ethyl acetate-cyclohexane solution(1:1,v/v) as the mobile phase.The separation of target compounds was carried out by a BEH C18 column and a gradient elution consisting of acetonitrile and 0.2% aqueous formic acid(v/v).The compounds were detected under the multiple reaction monitoring(MRM) mode and quantified with external standard method.This method was validated with respect to linearity,specificity,accuracy and precision.A linearity with correlation coefficient larger than 0.995 was achieved in the range of 0.5 to 50 ng m L~(-1).The average recoveries at the spiked levels of 1.0,5.0,and 10.0 μg kg~(–1) varied between 81.7% and 90.8%,with the relative standard deviations(n=5) ranged from 3.50% to 10.0%.The limit of quantification(LOQ) for 8 steroid hormones ranged from 0.2 to 1.5 μg kg~(-1).It was concluded that this method can be successfully applied for the determination of 8 steroid hormones in complicated matrices including high-fat fish tissues.     

Vegetation change of ecotone in west of Northeast China plain using time-series remote sensing data

Multi-temporal series of satellite SPOT-VEGETATION normalized difference vegetation index (NDVI) and normalized difference water index (NDWI) data from 1998 to 2007 were used for analyzing vegetation change of the ecotone in the west of the Northeast China Plain. The yearly and monthly maximal values,anomalies and change rates of NDVI and NDWI were calculated to reveal the interannual and seasonal changes in vegetation cover and vegetation water content. Linear regression method was adopted to characterize the trends in vegetation change. The yearly maximal NDVI decreased from 0.41 in 1998 to 0.37 in 2007,implying the decreasing trend of vegetation activity. There was a significant decrease of maximal NDVI in spring and summer over the study period,while an increase trend was observed in autumn. The vegetation-improved regions and vegetation-degraded regions occupied 17.03% and 20.30% of the study area,respectively. The maximal NDWI over growing season dropped by 0.027 in 1998–2007,and about 15.15% of the study area showed a decreasing trend of water content. Vegetation water stress in autumn was better than that in spring. Vegetation cover and water content variations were sensitive to annual precipitation,autumn precipitation and summer temperature. The vegetation degradation trend in this ecotone might be induced by the warm-drying climate especially continuous spring and summer drought in the recent ten years.     

Iron Regulation of Wetland Vegetation Performance Through Synchronous Effects on Phosphorus Acquisition Efficiency

Iron-rich groundwater flowing into wetlands is a worldwide environmental pollution phenomenon that is closely associated with the stability of wetland ecosystems. Combined with high phosphorus(P) loading from agricultural runoff, the prediction of the evolution of wetland vegetation affected by compound contamination is particularly urgent. We tested the effects of anaerobic iron-rich groundwater discharge in a freshwater marsh by simulating the effect of three levels of eutrophic water on native plants(Glyceria spiculosa(Fr. Schmidt.) Rosh.). The management of wetland vegetation with 1–20 mg/L Fe input is an efficient method to promote the growth of plants, which showed an optimum response under a 0.10 mg/L P surface water environment. Iron-rich groundwater strongly affects the changes in ecological niches of some wetland plant species and the dominant species. In addition, when the P concentration in a natural body of water is too high, the governance effect of eutrophication might not be as expected. Under iron-rich groundwater conditions, the δ~(13)C values of organs were more depleted, which can partially explain the differences in δ~(13)C in the soil profile. Conversely, the carbon isotope composition of soil organic carbon is indicative of past changes in vegetation. The results of our experiments confirm that iron-rich groundwater discharge has the potential to affect vegetation composition through toxicity modification in eutrophic environments.     

An integrated approach to assess the ecological and chemical status of lakes with HOD/AHOD: a case study of two lakes

The thermal strati?cation of the lakes impedes the transfer of atmospheric oxygen into the lower layers of the lake. In lakes which are af fected by diverse anthropogenic in?uences, the increasing organic matter amounts lead to a sharp decrease in hypolimnetic oxygen amounts, aided by thermal strati?cation,and anaerobic conditions arise. The determination of hypolimnetic oxygen demand(HOD) and areal hypolimnetic oxygen demand(AHOD) and their monitoring represent an integrated approach to investigate the oxygenation of lakes, the nutrient conditions, and the physicochemical dynamics. In this study, two lakes dif fering by size and af fected by dif ferent anthropogenic sources, are investigated in this respect. At?rst, bathymetric studies were conducted to determine the depth, surface area, and volume relationships.Then, based on monitoring studies conducted in 2013 and 2014, the thermal strati?cation dynamics and layer properties were established using the relative thermal resistance to mixing(RTRM) index based on temperature and density pro?les. Following this, the oxygen depletion rates were determined by oxygen and temperature pro?ling in the hypolimnion. For the years of 2013 and 2014, the AHOD values for the Borabey Pond which is far from anthropogenic in?uences, were found to be 0.848 and 0.569 g O 2/(m·d), respectively.The AHOD values for the Porsuk Reservoir which was overburdened for years by industrial and domestic pollution were found to be 4.263 and 5.099 g O 2/(m·d), larger than its counterpart by almost sevenfold. The HOD and AHOD monitoring can be considered to a valuable tool for assessing the ecological and chemical status of lakes within Annex 5 of the Water Framework Directive and as an integrated approach to assess and monitor the status of lakes.