[Fluorescence discrimination technique for phytoplankton based on the wavelet analysis].

Daubechies7 (db7) wavelet was selected to decompose the 3-D fluorescence spectra of 27 species of phytoplankton belonging to 22 generas of 6 divisions found in major lakes, then the scale vectors and time-series vectors were obtained as candidates for feature spectra. The third scale vector (Ca3) of db7 was chosen as feature spectra by Bayesian discriminant analysis, and the reference spectra were obtained via hierarchical cluster analysis to feature spectra. Based on the above data, a fluorescence discrimination technique was developed by multiple linear regression resolved by non-negative least squares. For single species algae cultures, the average correct discrimination ratio (CDR) was 98.6%, with the average relative content of 90.8% at division level. Furthermore, the noise immunity of reference spectra was tested by adding noise at different proportions. For the dominant division of laboratory mixed samples, the average CDR was 97.0%, with the average relative content of 67.7% at division level, and the average CDR of subdominant division was 90.7%, with the average relative content of 32.3%. The results showed that the technique is feasible to some extent.

[Estimation of the flux of inorganic nitrogen flowing into the East China Sea].

The flux of inorganic nitrogen flowing into the East China Sea was estimated based on the systematic analysis of all the pollution sources from 1980-2005. The result showed that the flux of inorganic nitrogen had been increasing from the early 1980s to the early 21st century. In detail, the flux was about 8.8 x 10(5) t x a(-1) in the early 1980s, and increased to about 2.6 x 10(6) t x a(-1) in the early of 21st century. The annual increasing rate was about 4.3%, and the mean flux was 1.8 x 10(6) t x a(-1). The flux of inorganic nitrogen of Yangtze River had also been increasing from early 1980s to the early 21st century. In detail, the flux was 4.0 x 10(5) t x a(-1) in the early 1980s, and increased to about 6.2 x 10(5) t x a(-1) in the middle 1980s, and was then kept at this value to the end of 1980s. After that, the flux value increased quickly from the early 1990s to 1.8 x 10(6) t x a(-1) in the early 21st century. Of all the sources, the proportion of land-source inorganic nitrogen was the largest, which was about 79%, among which, the river-source, the sewage-source and the non-point source accounted for 73%, 4% and 2%, respectively. Besides the land-source, the air-source and the mariculture-source accounted for 18% and 3%. The proportion of flux of Yangtze River in the river source was 76.5%.

[Studies on micelle behaviors of sophorolipid biosurfactant by steady-state fluorescence probe method].

The critical micelle concentration (CMC) of sophorolipid was determined by steady-state fluorescence probe method in which pyrene was used as fluorescence probe. Meanwhile, the changes in the sophorolipid CMC were examined in the addition of NaCl and aliphatic alcohol, respectively. The results showed that the CMC of sophorolipid was 1.3 x 10(-4) mol x L(-1) and had a slight decrease as NaCl concentration increased. However, the CMC of sophorolipid appeared to increase in the presence of aliphatic alcohol. The aggregation numbers and the micelle size of sophorolipid solution were investigated by fluorescence quenching and laser light scattering method. The aggregation numbers were 4-8 with the concentrations of sophorolipid in the range of 4 -8 CMC and the micelle mean diameter for sophorolipid with the concentration of 6 CMC was about 90 nm. These experimental results indicated that the micelle should be incompact.

[Research on the 3D discrete fluorescence spectrum technique for differentiation of phytoplankton population].

The present research was targeted to develop a fluorescence analyser for phytoplankton population which uses a series of LEDs as the light source. So the 3D discrete fluorescence spectra with 12 excitation wavelengths (400, 430, 450, 460,470, 490, 500, 510, 525, 550, 570 and 590 nm) were determined by fluorescence spectrophotometer for 43 phytoplankton species. Then, the wavelet, Daubechies-7 (Db7), and Bayes Classifier were applied to extract the characteristics for each classes from the 3D discrete fluorescence spectra. Lastly, the fluorescence differentiation method for phytoplankton populations was established by multivariate linear regression and non-negative least squares, which could differentiate phytoplankton populations at the levels of both divisions and genus. This method was tested: for simulatively mixed samples(the dominant species accounted for 70%, 80%, 90% and 100% of the gross biomass, respectively) from 32 red tide algal species, and the correct discrimination rates at the level of genus were 67.5%, 75.8%, 81.4% and 79.4%, respectively. For simulatively mixed samples (the dominant divisions algae accounted for 50%, 75% and 100% of the gross biomass, respectively) from 43 algal species, the discrimination rates at the level of division were 95.2%, 99.7% and 91.9% with average relative content of 38.1%, 63.2% and 90.5%, respectively.

Significant fluorescence enhancement by supramolecular complex formation between berberine chloride and cucurbit(n=7)uril and its analytical application.

The supramolecular interaction of cucurbit(n=7)uril (Q[7]) with berberine chloride (BER) has been studied in aqueous solution at pH 2.0 and room temperature by spectro-fluorimetry. The association constant of the complex was 2.07 × 10(6) L mol(-1) calculated by using a nonlinear least squares method. (1)H NMR spectra confirmed that a 1:1 stable complex is formed between Q[7] and BER. This work proposes a possible interaction mode, in which the guest BER is incorporated inside the hydrophobic cavity of the host Q[7] via the isoquinoline ring part of the guest molecule. Based on a significant enhancement of the fluorescence intensity of this supramolecular complex, a spectrofluorimetric method with high sensitivity and selectivity has been developed for the determination of BER in aqueous solution in the presence of Q[7]. The linear range of the method was from 7.43 to 11.2 × 10(3) ng mL(-1)with the detection limit 4.2 ng mL(-1). There was no interference from the compounds normally used in tablets, serum or urine constituents. The proposed method was applied to the determination of BER in tablets, serum and urine samples with satisfactory results and good consistency with those obtained by the pharmacopoeia method. This shows that it has promising potential for therapeutic drug monitoring and pharmacokinetics and for clinical application.

[Toxic effects of chloride tributyltin on the predominant phytoplankton species of China coastal sea].

Toxic effects of tributyltin (TBT) on the growth, peak diameter and photosynthetic activity (F(v)/F(m)) of fifteen predominant phytoplankton species in China coastal sea of Leptocylindru danicus, Skeletonema marinoi, Asterionella japonica, Bacilaria paxillifera, Chaetoceros debilis, Chaetoceros socialis, Thalassiosira, Amphidinium carterae, Karenia mikimotoi, Prorocentrum minimum, Gymnodinium sp., Scrippsiella trochoidea, Gymnodinium simplex, Prorocentrum triestinum and Heterosigma akashiwo were systematically studied. 96 h-EC50 of TBT were 136.42, 3.55, 3.15, 2.95, 1.90, 1.08, 0.70, 21.60, 7.71, 4.77, 3.81, 2.04, 0.53, 0.51, 1.76 microg/L, respectively. The no detectation effect concentration were 112.62, 1.19, 1.17, 3.70, 0.95, 1.07, 0.80, 6.28, 1.84, 0.99, 3.73, 1.63, 0.23, 0.16, 1.76 microg/L, respectively. The results indicated that the toxic effects of TBT were very different among different phytoplankton species and the resistance of Leptocylindru danicus was stronger than those of other microalgae. The TBT not only inhibited the growth of phytoplankton, but also changed the peak diameter and decreased the photosynthetic activity (F(v)/F(m)). Besides that, we infer that there may have a relationship between the toxicity of TBT and the composition of predominant phytoplankton population in harbour.

[Research on the 3D fluorescence spectra differentiation of phytoplankton by coiflet2 wavelet].

In the present paper, the authors utilize the wavelet base function coiflet2 (coif2) to analyze the 3D fluorescence spectra of 37 phytoplankton species belonging to 30 genera of 7 divisions, and these phytoplankton species include common species frequently causing harmful algal blooms and most predominant algal species in the inshore area of China Sea. After the Rayleigh and Raman scattering peaks were removed by the Delaunay triangulation interpolation, the fluorescence spectra of those phytoplankton species were transformed with the coiflet2 wavelet, and the scale vectors and the wavelet vectors were candidate for the feature spectra. Based on the testing results by Bayesian analysis, the 3rd scale vectors were the best feature segments at the division level and picked out as the fluorescence division feature spectra of those phytoplankton species, and the group of the 3rd scale vectors, the 2nd and 3rd wavelet vectors were the best feature segments at the genus level and chosen as the fluorescent genus feature spectra of those phytoplankton species. The reference spectra of those phytoplankton species at the division level and that at the genus level were obtained from these feature spectra by cluster analysis, respectively. The reference spectra base for 37 phytoplankton species was composed of 107 reference spectra at the division level and 155 ones at the genus level. Based on this reference spectra base, a fluorometric discriminating method for phytoplankton populations was established by multiple linear regression resolved by the nonnegative least squares. For 1 776 samples of single phytoplankton species, a correct discriminating rate of 97.0% at genus level and 98.1% at division level can be obtained; The correct discriminating rates are more than 92.7% at the genus level and more than 94.8% at the division level for 384 mixed samples from two phytoplankton species.

[Fluorescence characterization of dissolved organic matter in East China sea].

Sea water samples were collected in the East China Sea in March and April, 2005, and three-dimensional fluorescence of dissolved organic matter was measured by fluorescence excitation-emission matrix spectroscopy. The position, number and intensity of fluorescence peak in the spectra and the relations of the peaks were analyzed to determine the type, distribution and origin of the fluorescence dissolved organic matter. Seven types of fluorescence peaks were detected from the samples. There are protein-like fluorescence peaks B with Exmax/Emmax = 275/300 nm, D with Exmax/Emmax = 225/295-305 nm, T with Exmax/Emmax = 280/345 nm, and S with Exmax/Emmax = 225-240/320-350 nm, two humic-like peaks A with Exmax/Emmax = 250-255/410-455 nm and C 335-345/410-440 nm, and marine humic peak M with Exmax/Emmax = 305 nm/400-420 nm. Peaks B, S and A appeared in all surveyed area. Peaks T and D appeared in the north of the surveyed area. Peaks M and C only appeared in a few stations. In the surface layer, the source of the fluorescence dissolved organic matter might be the fresh water outflow of the Yangtze River, while the fluorescence dissolved organic matter in the middle layer had double sources from the Yangtze River and the phytoplankton. The good correlationships of different fluorescence peaks showed the same source or some relationship between the protein-like and the humic-like fluorescence dissolved organic matter.

[Role of irradiance on the seasonality of Skeletonema costatum Cleve blooms in the coastal area in East China Sea].

With field culture experiments and model calculations, the role of light on why in both spring and summer Skeletonema costatum Cleve blooms can occur in the coastal area in East China Sea were studied. The results show that the growth optimal light intensity (I(opt)) of S. costatum increases gradually to a maximum of 121.6 W x m(-2) with temperature (t) up to 25 degrees C, and a rapid decline in I(opt) accompanies further increases in t beyond 25 degrees C. Such an asymmetric inverse V-shape I(opt)-t curve can be well described by Blanchard's equation. By this equation, I(opt) of each month was calculated based on monthly sea surface temperature, and though changes about 2 months later than light intensity above sea surface (I(0)), I(opt) is closely correlated with seasonal I(0) and transparency of sea water (R2 = 0.907 +/- 0.115, p < 0.001), which indicates that S. costatum adapts to the seasonality of light in sea water in the coastal area by varying I(opt) seasonally. Such an adaptive strategy leads to thick (about 6 m) light-optimal water layers for growth from early spring to late summer. The high adaptability to light and light optimum characteristics in sea water is one of the important reasons for S. costatum blooms in the both seasons in the coastal area in ECS.

[Effects of irradiance on blooms of the dinoflagellate Prorocentrum donghaiense Lu in the coastal area in East China Sea].

With field culture experiments and model calculations, the natural-light-dependent growth and the optimal light layers in sea water for growth of red tide dinoflagellate Prorocentrum donghaiense Lu were studied in order to analyze the role of light on algal blooms in the coastal area in East China Sea in spring. The results show that the relationship of growth and light can be well described by Steele's equation, and the optimal light intensity (Iopt) of P. donghaiense is (38.2 +/- 3.8) W x m(-2), which is lower than Iopt for several other red tide algae (40-133 W x m(-2)), meaning that P. donghaiense may have an advantage when developing blooms in turbid environments where solar irradiance is easily attenuated. The optimal light layers for P. donghaiense growth are thicker offshore than inshore, and the thickness of optimal light layers in the subsurface water usually in 3-15 m in depth is about 5-10 m in the red tide area. The trade-off of light and nutrient fitness results in blooms in the so-called red tide area, and the light-optimum characteristic of the subsurface water is an important factor for the subsurface bloom development in spring.

[Excitation-emission-matrix spectral property of dissolved organic matter in seawater of Jiaozhou Bay, China].

Excitation-emission matrix spectroscopy (EEMs) was applied to measure the fluorescence properties of dissolved organic matter (DOM) in seawater collected from Jiaozhou Bay in June, 2005. The study areas include central Bay area, receptor areas of Dagu River, Licun River and Haibo River. The aim was to investigate the influence of discharges from several typical rivers to the receiving seawater. Dagu River discharges mostly freshwater, while the other two rivers discharges include more wastewaters from factories and sewages treatments. The results indicate that there is a redshift of the position of humic-like fluorescence peaks due to the discharges from sewages and wastewaters from factories. The intensities of protein-like and humic-like peaks were the strongest in the samples from area near Haibo River and Licun River, while the weakest fluorescence intensities of both protein-like and humic-like were observed in the samples collected from central bay area. This suggests that river inputs are the mains sources of DOM in seawater of Jiaozhou Bay, while DOM produced by bioactivity in situ dominates those in central Bay area. The relationships of protein-like and humic-like fluorescence intensities vary depending on the water samples. The DOM in seawater of Jiaozhou Bay may be polluted by protein-like organic matter produced by human activity, as indicated by higher ratios of protein-like and humic-like fluorescence intensities. The results suggest the potential utility of EEMs technique for trace DOM and land-based pollution sources.

Spatial distribution of dissolved Pb, Hg, Cd, Cu and As in the Bohai Sea.

Characteristics of the spatial distribution of selected dissolved heavy metals were analyzed during large scale surveys from August 12 to 25, 2003 in the Bohai Sea. Dissolved Pb was the only element with average concentrations higher than the grade-one sea water quality standard of China. The spatial distribution of dissolved Pb in surface water was similar to those of Cd, Cu and As, where the isopleths generally indicated decreasing values from the bays to the central areas. Only for Hg did the high concentrations not only appear in Liaodong Bay, Bohai Bay and Laizhou Bay, but also in the Central Area, viz. not only in inshore but also in offshore areas. Vertical distributions of dissolved Pb, Cd, Cu and As were largely uniform, while that of dissolved Hg increased with depth. We infer that the input of pollutants from land was the main influencing factor for the detected distribution patterns of dissolved heavy metals, followed by the dynamics of sea water, release from bottom sediments and biochemical processes. Comparing with historical data, average concentrations of dissolved heavy metals appear to decline in recent years.

The application and comparison of several chemometric methods of excitation-emission matrix spectra in studying the interactions of metal complexes with DNA.

The interactions of fs DNA and two metal complexes [Cu(phen)SO4] x 2 H2O and [Ni(phen)SO4] x 2 H2O were explored by several chemometric methods, including parallel factor (PARAFAC), singular value decomposition-least squares (SVD-LS), and singular value decomposition-nonnegative least squares (SVD-NNLS) of excitation-emission matrix spectra (EEMs). The applications of SVD-LS and SVD-NNLS in this domain have been discussed. Rayleigh scatter part is avoided by ordered zero and reconstructed by linear interpolation. The importance of avoiding Rayleigh scatter has also been discussed. All the three methods do well in qualitative analysis. SVD-LS does best in present small changes of ethidium bromide (EB). In order to get accurate results, PARAFAC and SVD-NNLS can be utilized together in quantitative analysis. All the three chemometric methods indicate that the DNA binding modes of [Cu(phen)SO4] x 2 H2O are hydrogen bond effect and intercalation, while intercalation is the only DNA binding mode for [Ni(phen)SO4] x 2 H2O. These results are verified by the electronic absorption and emission fluorescence spectra. Just like PARAFAC, both SVD-LS and SVD-NNLS are proven to be convenient and convincing in studying the interactions between nucleic acids and complexes.

[Exchange rates of dissolved nutrients at the sediment-water interface before and after diatom bloom in the East China Sea in spring].

Exchange rates of dissolved nutrients at the sediment-water interface were studied before and after diatom bloom from March to May in the East China Sea in 2005. Intact sediment cores which were sampled at 10 sites were incubated on board, and exchange rates were calculated in the function method. It was shown that sediment absorbed NO3(-) -N before bloom with the exchange rate ranging from - 1.33 mmol/(m2 d) to - 0.68 mmol/(m2 d). The sediment released NO3(-) -N after bloom with the exchange rate ranging from - 0.69 mmol/( m2 d) to 0.82 mmol/( m2 d). NH4(+) -N was almost released from sediment with the exchange rate ranging from -0.65 mmol/(m d) to 1.46 mmol/( m2 d), and the rate was higher before bloom than that after bloom. NO2(-) -N moved into sediment all along except at Zc17 with the exchange rate ranging from - 0.09 mmol/(m2 d) to 0.05 mmol/(m2 d), and the rate was a little higher after bloom than that before bloom. SiO3(2-) -Si was released from sediment all the time with the exchange rate ranging from 0.85 mmol/(m d) to 9.23 mmol/( m2 d), and the rate was higher after bloom than that before bloom. The sediment absorbed PO4(3-) -P before bloom with the exchange rate ranging from -0.06 mmol/(m2 d) to -0.01 mmol/(m2 d), but released PO4(3-) -P after bloom with the exchange rate ranging from 0 mmol/(m2 d) to 1.26 mmol/(m2 d). PO4(3-) -P and NO3(-) -N which were absorbed by sediment in spring restricted the diatom bloom, but their release from sediment after diatom bloom may make a great contribution to subsequent dinoflagellate bloom.

[Discrimination of Red Tide algae by fluorescence spectra and principle component analysis].

Fluorescence discrimination technology for 11 species of the Red Tide algae at genus level was constructed by principle component analysis and non-negative least squares. Rayleigh and Raman scattering peaks of 3D fluorescence spectra were eliminated by Delaunay triangulation method. According to the results of Fisher linear discrimination, the first principle component score and the second component score of 3D fluorescence spectra were chosen as discriminant feature and the feature base was established. The 11 algae species were tested, and more than 85% samples were accurately determinated, especially for Prorocentrum donghaiense, Skeletonema costatum, Gymnodinium sp., which have frequently brought Red tide in the East China Sea. More than 95% samples were right discriminated. The results showed that the genus discriminant feature of 3D fluorescence spectra of Red Tide algae given by principle component analysis could work well.

[Research on characterization analysis of synchronous fluorescence spectra of living phytoplankton].

The synchronous fluorescence spectra of five phytoplankton species growing under three temperatures (25, 20 and 15 degrees C)and three illuminations (7000, 4100 and 1100 Lx)were measured and processed by multinomial smoothness and autoscaling to obtain the characteristic spectra. Principal component analysis was used to obtain standard spectra. The analysis shows that at different temperatures, the characteristic spectra of Skeletonema costatuma, Isochrysis galbana, and Platymonas helgolanidica show high similarities, while the spectra similarities of Alexandrium tamarense and Gymnodinium stein are not as good as the above three species. The standard spectrum of Skeletonema costatuma, which belongs to Bacillariophyta, is quite different from those of Alexandrium tamarense and Gymnodinium stein, which belong to Dinophyta.

[Synthesis and DNA binding spectroscopic studies of Cu(II)-HAsp-Im complexes].

Two novel complexes of [Cu(HAsp)ImH2O]SO4 x 4 H2O and [Cu(Asp)Im(OH)] x 4 H2O (HAsp = Aspartic acid molecule, Asp = Aspartic acid ion, Im = Imidazole) were synthesized and characterized by elemental analysis, IR spectroscopy and TG-DTA. The interactions of the complexes with sperm DNA were studied by electronic absorption and fluorescence spectra. Results showed that the reactions of the two complexes with DNA are obviously different: for [Cu(HAsp)ImH2O]SO4 x 4 H2O, it is intercalation companied by electrostatic effect, while [Cu(Asp)Im(OH)] x 4 H2O mainly cooperates with the nitrogen atom of the DNA base pair, which induces the breakage of DNA double helix. The reasons for these differences in their DNA binding modes were also discussed.

[Ecological effect of No.0 diesel water accommodated fraction on marine algae].

With batch culture experiments in field and laboratory, the ecological effect of No. 0 diesel water accommodated fraction on marine algae was studied. A growth model of marine algae under grazing pressure and a model of growth effect on marine algae with different doses No.0 diesel water accommodated fraction were proposed. Based on the model and experiments, the growth effect of No.0 diesel water accommodated fraction on marine algae was studied. The results show that, the growth model of marine algae under grazing pressure is more suited for the marine ecological system than Logistic model. And the final biomass (B(f)) of marine algae with different doses No.0 diesel water accommodated fraction was calculated by the model with none-linear fitting software. The results also show that, under the field and laboratory conditions, lower doses No.0 diesel water accommodated fraction promotes the growth of marine algae, and the most promoting ratio are 180% and 120% respectively, however, higher doses hardly promotes but bates the growth of marine algae.

[Seasonal variation and spatial distribution of nutrients and their relationships with harmful algal blooms in coastal area of the East China Sea].

Based on the data from four cruises that carried out in the Changjiang River estuary and its adjacent areas from 2002-04 to 2003-02, the seasonal variation and spatial distribution of nutrients were analyzed, and the relationship between nutrients condition and the harmful algal blooms (HABs) was also discussed. Results showed that the annual average concentration of nutrient was (17.93 +/- 2.46) micromol x L(-1) for DIN, (0.59 +/- 0.11) micromol x L(-1) for PO4(3-) -P, (15.34 +/- 3.23) micromol x L(-1) for SiO3(2-) -Si, and the study area was in the state of eutrophication. The average concentration of nutrient showed a remarkable seasonal fluctuation with the higher value in autumn and winter and lower value in spring and summer. The spatial distribution of the nutrients was typically such that the concentrations of DIN, PO4(3-) -P and SiO3(2-) -Si decreased from inshore to offshore area, and the contours generally decreased rapidly in down-coast directions. DIN and SiO3(2-) -Si of the study area were largely contributed by Changjiang River diluted water and other terrigenous inputs, while PO4(3-) -P mainly by Changjiang River diluted water and the Taiwan Warm Current. The following HABs showed that its occurrences were usually laid in the areas with higher nutrients and lower total suspended particles (TSP).

[Study on the characters of phytoplankton chlorophyll fluorescence excitation spectra based on fourth-derivative].

Chlorophyll fluorescence excitation spectra of six phytoplankton species, belonging to Bacillariophyta and Dinophyta, were dealt by fourth-derivative analysis with the Matlab program. The results show that between 350 nm and 550 nm six fluorescence peaks were found in the fourth-derivative spectra, which are representatives of non-pigments, chlorophylls and carotenoides respectively. The method makes Bacillariophyta and Dinophyta more distinguishable when the fourth-derivative spectra are compared with the chlorophyll fluorescence excitation spectra. It can be used not only to discriminate the two groups of algaes, but also to reduce the effect of noise. The fluorescence peaks in the fourth-derivative spectra are proved to be stable.