Rongguo Su

Investigation of seasonal variability of CDOM fluorescence in the southern changjiang river estuary by EEM-PARAFAC

The southern Changjiang River Estuary has attracted considerable attention from marine scientists because it is a highly biologically active area and is biogeochemically significant. Moreover, land-ocean interactions strongly impact the estuary, and harmful algal blooms (HABs) frequently occur in the area. In October 2010 and May 2011, water samples of chromophoric dissolved organic matter (CDOM) were collected from the southern Changjiang River Estuary. Parallel factor analysis (PARAFAC) was used to assess the samples’ CDOM composition using excitation-emission matrix (EEM) spectroscopy. Four components were identified: three were humic-like (C1, C2 and C3) and one was protein-like (C4). Analysis based on spatial and seasonal distributions, as well as relationships with salinity, Chl a and apparent oxygen utilization (AOU), revealed that terrestrial inputs had the most significant effect on the three humic-like Components C1, C2 and C3 in autumn. In spring, microbial processes and phytoplankton blooms were also important factors that impacted the three components. The protein-like Component C4 had autochthonous and allochthonous origins and likely represented a biologically labile component. CDOM in the southern Changjiang River Estuary was mostly affected by terrestrial inputs. Microbial processes and phytoplankton blooms were also important sources of CDOM, especially in spring. The fluorescence intensities of the four components were significantly higher in spring than in autumn. On average, C1, C2, C3, C4 and the total fluorescence intensity (TFI) in the surface, middle and bottom layers increased by 123%–242%, 105%–195%, 167%–665%, 483%–567% and 184%–245% in spring than in autumn, respectively. This finding corresponded with a Chl a concentration that was 16–20 times higher in spring than in autumn and an AOU that was two to four times lower in spring than in autumn. The humification index (HIX) was lower in spring that in autumn, and the fluorescence index (FI) was higher in spring than in autumn. This result indicated that the CDOM was labile and the biological activity was intense in spring.

Identifying phytoplankton in seawater based on discrete excitation-emission fluorescence spectra.

The feasibility of utilizing discrete excitation‐emission spectra (DEEMs) to identify dominant groups of phytoplankton at both the genus and division levels was investigated. First, the characteristics of in vivo DEEMs were extracted using Coif2 wavelet. Second, optimal characteristic spectra of scale vectors (SOCS) and time‐series vectors (TOCS) were selected by Fisher linear discriminant analysis (FLDA). Third, the SOCS and TOCS were sorted using hierarchical cluster analysis (HCA), and a two‐rank database was established according to their discrimination ability. Fourth, the discrimination of phytoplankton was established by nonnegative least squares (NNLS). For single‐species samples, the correct identification ratios (CIRs) were 62.9%–100% at the genus level and 95.1%–100% at the division level. The dominant species in the mixtures had corresponding CIRs of 87.5% and 97.9%, and 23 dominant species were correctly identified. Prorocentrum donghaiense D. Lu, Thalassiosira nordenskioeldi Cleve, Chaetoceros socialis Lauder (bloom‐forming species with a density of about 107 cell·L−1), and Skeletonema costatum (Grev.) Cleve (a dominant species with a density of 104–106 cell·L−1 in seawater) were identified at the genus level. Other dominant species in seawater were identified at the division level if their density was 105–106 cell·L−1.

The assessment of the spatial and seasonal variability of chromophoric dissolved organic matter in the Southern Yellow Sea and the East China Sea.

Samples of chromophoric dissolved organic matter (CDOM) from the Southern Yellow Sea (SYS) and the East China Sea (ECS) were evaluated by fluorescent Excitation Emission Matrix (EEM) combined with Parallel Factorial Analysis (PARAFAC). Three terrestrial humic-like components (C1, C2 and C3) and one autochthonous protein-like component (C4) were identified. As for seasonal variations, CDOM displayed the following order on the whole: summer>spring>autumn. The C1, C2 and C3 components were mainly dominated by terrestrial inputs and their spatial distributions and temporal variations also can be influenced by primary productivity of phytoplankton, microbial activities and photobleaching. C4 was produced by phytoplankton and microorganisms and consumed by marine bacteria, and besides its distribution was attributed to the influence of riverine inputs. Terrestrial inputs were the dominant sources of CDOM in the SYS and ECS.

Characterization of Chromophoric Dissolved Organic Matter (CDOM) in the Bohai Sea and the Yellow Sea Using Excitation-Emission Matrix Spectroscopy (EEMs) and Parallel Factor Analysis (PARAFAC)

Compositions and concentrations of chromophoric dissolved organic matter (CDOM) have been determined in the Bohai Sea (BS) and the Yellow Sea (YS) in the summer and autumn in 2013 by excitation and emission matrix spectroscopy (EEMs) and parallel factor analysis (PARAFAC). The PARAFAC model identified three humic-like components (C1, C2, and C3) and one protein-like component (C4). CDOM exhibited higher fluorescence intensities in the coastal areas in both the summer and autumn. However, its distribution patterns were different in the two seasons. Based on spatial and seasonal distributions of four components, as well as correlations with salinity, chlorophyll a (Chl-a), and apparent oxygen utilization (AOU), the following assignments were made. The C1, C2, and C3 components were mainly dominated by terrestrial inputs and influenced by the primary productivity of phytoplankton in the summer as well. C4 was assigned to terrestrial and autochthonous origins and most likely represented a biologically labile component. Terrestrial inputs were the dominant source of CDOM in the BS and YS. The humification index (HIX) and biological index (BIX) suggested that CDOM in the BS was more stable than that in the YS, which had an increase in autochthonous production, and in the summer, CDOM was less stable with a higher CDOM autochthonous production compared with that in the autumn.

Real-time eutrophication status evaluation of coastal waters using support vector machine with grid search algorithm

The development of techniques for real-time monitoring of the eutrophication status of coastal waters is of great importance for realizing potential cost savings in coastal monitoring programs and providing timely advice for marine health management. In this study, a GS optimized SVM was proposed to model relationships between 6 easily measured parameters (DO, Chl-a, C1, C2, C3 and C4) and the TRIX index for rapidly assessing marine eutrophication states of coastal waters. The good predictive performance of the developed method was indicated by the R2 between the measured and predicted values (0.92 for the training dataset and 0.91 for the validation dataset) at a 95% confidence level. The classification accuracy of the eutrophication status was 86.5% for the training dataset and 85.6% for the validation dataset. The results indicated that it is feasible to develop an SVM technique for timely evaluation of the eutrophication status by easily measured parameters.

Assessing Phytoplankton Using a Two-Rank Database Based on Excitation-Emission Fluorescence Spectra

The feasibility of using a two-rank database of reference spectra based on in vivo fluorescence excitation-emission matrix (EEMs) spectra to assess dominant groups of phytoplankton was explored. Twenty-six species belonging to 20 genera of seven divisions were studied. First, fluorescent characteristics of these EEMs were extracted using Daubechies-7 wavelet analysis. Second, the optimal characteristic spectra of scale vectors (SOCS) and time-series vectors (TOCS) were selected; phytoplankton of different genera were classified using Fisher linear discriminant analysis. Third, SOCS and TOCS reference spectra databases were obtained by hierarchical cluster analysis. Using non-negative least squares as the method to assess the phytoplankton, a two-rank reference spectra database was established according to their respective ability to identify the 2818 single-species samples. Using this fluorimetric technique, the correct identification rates (CIRs) for single-species samples were 88.8–100% at the genus level and 98.8–100% at the division level. Dominant species in the 465 laboratory mixtures had corresponding CIRs of 85.6% and 96.1%. Moreover, 15 of the 19 species used as dominants were correctly identified at the genus level. In 27 natural seawater samples, Prorocentrum donghaiense, Thalassiosira nordenskioldi, and Chaetoceros socialis (bloom-forming species with a density of about 107 cell L−1), and Alexandrium sp. (dominant species with abundance of about 106 cell L−1) were qualitatively identified at the genus level; other dominant species, with densities of 105 to 106 cell L−1, were identified at the division level. The quantitative identification was relatively poor in the natural water samples, and several potential resolutions, including trying both new measuring methods and calculating methods, for future study are given.

Discrimination of marine algal taxonomic groups based on fluorescence excitation emission matrix, parallel factor analysis and CHEMTAX

An in vivo three-dimensional fluorescence method for the determination of algae community structure was developed by parallel factor analysis (PARAFAC) and CHEMTAX. The PARAFAC model was applied to fluorescence excitation-emission matrix (EEM) of 60 algae species belonging to five divisions and 11 fluorescent components were identified according to the residual sum of squares and specificity of the composition profiles of fluorescent. By the 11 fluorescent components, the algae species at different growth stages were classified correctly at the division level using Bayesian discriminant analysis (BDA). Then the reference fluorescent component ratio matrix was constructed for CHEMTAX, and the EEM-PARAFAC-CHEMTAX method was developed to differentiate algae taxonomic groups. The correct discrimination ratios (CDRs) when the fluorometric method was used for single-species samples were 100% at the division level, except for Bacillariophyta with a CDR of 95.6%. The CDRs for the mixtures were above 94.0% for the dominant algae species and above 87.0% for the subdominant algae species. However, the CDRs of the subdominant algae species were too low to be unreliable when the relative abundance estimated was less than 15.0%. The fluorometric method was tested using the samples from the Jiaozhou Bay and the mesocosm experiments in the Xiaomai Island Bay in August 2007. The discrimination results of the dominant algae groups agreed with microscopy cell counts, as well as the subdominant algae groups of which the estimated relative abundance was above 15.0%. This technique would be of great aid when low-cost and rapid analysis is needed for samples in a large batch. The fluorometric technique has the ability to correctly identify dominant species with proper abundance both in vivo and in situ.

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 mL−1. After purification, the isolated bioactive substances (the organic extract C) exhibited much higher antialgal and antilarval activities with EC50 of 12.9 μg mL−1 against Skeletonema costatum and LC50 of 15.1 μg mL−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.

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.

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 EC50 and LC50 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 EC50 value of 7.4 μg mL−1 against S. costatum and LC50 value of 9.7 μg mL−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.