Acute toxicity of live and decomposing green alga Ulva (Enteromorpha) prolifera to abalone Haliotis discus hannai

From 2007 to 2009, large-scale blooms of green algae (the so-called “green tides”) occurred every summer in the Yellow Sea, China. In June 2008, huge amounts of floating green algae accumulated along the coast of Qingdao and led to mass mortality of cultured abalone and sea cucumber. However, the mechanism for the mass mortality of cultured animals remains undetermined. This study examined the toxic effects of Ulva (Enteromorpha) prolifera, the causative species of green tides in the Yellow Sea during the last three years. The acute toxicity of fresh culture medium and decomposing algal effluent of U. prolifera to the cultured abalone Haliotis discus hannai were tested. It was found that both fresh culture medium and decomposing algal effluent had toxic effects to abalone, and decomposing algal effluent was more toxic than fresh culture medium. The acute toxicity of decomposing algal effluent could be attributed to the ammonia and sulfide presented in the effluent, as well as the hypoxia caused by the decomposition process.     

Ulva and Enteromorpha (Ulvaceae, Chlorophyta) from two sides of the Yellow Sea: analysis of nuclear rDNA ITS and plastid rbcL sequence data

Ulvacean green seaweeds are common worldwide; they formed massive green tides in the Yellow Sea in recent years, which caused marine ecological problems as well as a social issue. We investigated two major genera of the Ulvaceae, Ulva and Enteromorpha, and collected the plastid rbcL and nuclear ITS sequences of specimens of the genera in two sides of the Yellow Sea and analyzed them. Phylogenetic trees of rbcL data show the occurrence of five species of Enteromorpha (E. compressa, E. flexuosa, E. intestinalis, E. linza and E. prolifera) and three species of Ulva (U. pertusa, U. rigida and U. ohnoi). However, we found U. ohnoi, which is known as a subtropical to tropical species, at two sites on Jeju Island, Korea. Four ribotypes in partial sequences of 5.8S rDNA and ITS2 from E. compressa were also found. Ribotype network analysis revealed that the common ribotype, occurring in China, Korea and Europe, is connected with ribotypes from Europe and China/Japan. Although samples of the same species were collected from both sides of the Yellow Sea, intraspecific genetic polymorphism of each species was low among samples collected worldwide.     

Dynamic Diurnal Changes in Green Algae Biomass in the Southern Yellow Sea Based on GOCI Images

Macroalgae blooms of Ulva prolifera have occurred every summer in the southern Yellow Sea since 2007, inducing severe ecological problems and huge economic losses. Genesis and secular movement of green algae blooms have been well monitored by using remote sensing and other methods. In this study, green algae were detected and traced by using Geostationary Ocean Color Imager(GOCI), and a novel biomass estimation model was developed from the relationship between biomass measurements and previously published satellite-derived biomass indexes. The results show that the green algae biomass can be determined most accurately with the biomass index of green algae for GOCI(BIGAG), which is calculated from the Rsurf data that had been atmospherically corrected by ENVI/QUAC method. For the first time, dynamic changes in green algae biomass were studied over an hourly scale. Short-term biomass changes were highly influenced by Photosynthetically Available Radiation(PAR) and tidal phases, but less by sea surface temperature variations on a daily timescale. A new parameter of biomass changes(PBC), calculated by the ratio of the biomass growth rate to movement velocity, could provide an effective way to assess and forecast green tide in the southern Yellow Sea and similar areas.     

Molecular phylogenetic analysis of attached Ulvaceae species and free-floating <Emphasis Type="Italic">Enteromorpha</Emphasis> from Qingdao coasts in 2007

Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer (ITS) 1, 5.8 S, and ITS 2, the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007, including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations. The result supported the monophyly of all free-floating Enteromorpha samples, implying the unialgal composition of the free-floating Enteromorpha, and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades, suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season. Supported by NSFC (40506030); the Innovative Key Project of the Chinese Academy of Sciences (KZCX2-YW-209); Science & Technology Project of Qingdao City (06-2-2-12-JCH)     

Bacterial diversity in surface water of the Yellow Sea during and after a green alga tide in 2008

From May to August 2008, a large "green tide", consisting of the alga Ulva (Enteromorpha) prolifera, occurred in the Yellow Sea, China, affecting the local marine ecosystem and human activities. We investigated the influence of the green tide on the microbial community in the surface seawater, at four sites from July to August 2008, using bacterial 16S rRNA gene clone libraries. We sequenced 228 clones of unique patterns identified by restriction fragment length polymorphism (RFLP) techniques. The results show that 228 sequenced clones fell into six bacterial phyla:Proteobacteria, Bacteroidetes, Cyanobacteria, Verrucomicrobia, Actinobacteria, and Planctomycetes. Alphaproteobacteria (33%), Gammaproteobacteria (25%), Bacteroidetes (23%) and Cyanobacteria (9%) dominated the assemblage. Comparison between samples collected in July (during the tide) and those collected in August (after the tide) showed that, in the microbial community, diversities of Alphaproteobacteria and Cyanobacteria increased after the tide, while those of Gammaproteobacteria and Bacteroidetes decreased. These results indicate that the green tide influenced the growth of some bacteria, and provide information for further studies on the interactions and relationships between U. prolifera and the bacterial community. This study suggests that microbial community analysis is a good approach to monitoring green tides.     

A New Molecular Label Applied to the Study of the Yellow Sea Green Tide

From 2007 to 2017, large-scale green tides occurred every year in the Yellow Sea of China, and Ulva prolifera was the main species leading to the green tide. In this study, we used the Polymerase chain reaction and 3' Rapid-amplification of c DNA ends technique to amplify the nr DNA-LSU and IGS sequences in U. prolifera and one species of Blidingia. These techniques showed 3259 bp of nrD NA-LSU and 3388 bp of IGS in U. prolifera and 3282 bp nr DNA-LSU and 3059 bp IGS in Blidingia sp. At the same time, tandem repeats, short dyads, palindromic and multiple simple repeat sequences in the IGS sequence were found by analyzing the structure of the IGS sequence in U. prolifera and Blidingia sp. G + C contents of the IGS sequence in U. prolifera and Blidingia sp. were 52.42% and 53.09%, respectively. We divided the U. prolifera into two types according to the morphological characteristics. Although the specimens of U. prolifera from the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea have different morphologies, their ITS and IGS sequences are almost identical. Therefore, the main species of the green tides in the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea are the same and have the same origin.     

DNA barcoding assessment of green macroalgae in coastal zone around Qingdao,China

An assessment with assistance of DNA barcoding was conducted on green macroalgae in coastal zone around Qingdao, China, during the period of April–December, 2011. Three markers were applied in molecular discrimination, including the plastid elongation factor tufA gene, the internal transcribed spacer (ITS) region of the ribosomal cistron and rubisco large subunit gene 3′ regions (rbcL-3P). DNA barcoding discriminated 8 species, excluding species of genus Cladophora and Bryopsis due to failures in amplification. We ascertained and corrected 4 species identified by morphological methods for effectively assisting the classification. The gene tufA presented more advantages as an appropriate DNA marker with the strongest amplification success rate and species discrimination power than the other two genes. The poorest sequencing success largely handicapped the application of ITS. Samples identified by tufA and rbcL as Ulva flexuosa were clustered into the clade of U. prolifera by ITS in the neighbor-joining tree. Confusion with discrimination of the complex of U. linza, U. procera and U. prolifera (as the LPP complex) still existed for the three DNA markers. Based on our results, rbcL is recommended as a preferred marker for assisting tufA to discriminate green macroalgae. In distinguishing green-tide-forming Ulva species, the free-floating sample collected from the green tide in 2011 was proved to be identical with U. prolifera in Yellow Sea for ITS and rbcL genes. This study presents a preliminary survey of green macroalgae distributed in the coastal area around Qingdao, and proves that DNA barcoding is a powerful tool for taxonomy of green macroalgae.     

Spatiotemporal variation in community structure of marine benthic ciliates in the Yellow Sea during and after macroalgal and giant jellyfish blooms

The annual bloom of the green macroalgal Ulva prolifera from May through July since 2008 and another of giant jellyfish Nemopilema nomurai from June through September have been frequent events in the Yellow Sea. However, the patterns of benthic ciliate communities during and after the blooms are still not known. In combination with analyses of benthic environmental factors, we investigated the distribution and community composition of benthic ciliates in the Yellow Sea in July and November 2011. In July, ciliates had high standing crops and diversity in the northern Yellow Sea, and in the inshore area off the southern Shandong Peninsula, where large numbers of green macroalgae accumulated. In November, the abundance, biomass and diversity of ciliates were high in the sea areas off the Shandong Peninsula and Changjiang estuary, where a large quantity of jellyfish occurred in August. Neither the abundance nor the biomass had significant difference between seasons, or between different compartments of the Yellow Sea. The species number, and both Margalef and Shannon-Wiener indices of ciliates were all significantly higher in November than in July. In both seasons, prostomateans and karyorelicteans consistently constituted the first and second most important ciliate groups in biomass; and carnivorous ciliates constituted the primary feeding type in terms of biomass as well as species richness, followed by bacterivores, algivores and omnivores. Compared with that in June 2007 when no macroalgae occurred, the percentage of small-sized bacterivores (e.g. Metacystis spp., Euplotes spp. and scuticociliates) increased in July 2011. The proportion of carnivorous ciliates increased in November, and this increased dominance of carnivorous ciliates may be a response to the increase in predominance of heterotrophic nanoflagellates, which might in turn be ascribed to an effect of green macroalgal and giant jellyfish blooms in the Yellow Sea.     

Antioxidant system responses in two co-occurring green-tide algae under stress conditions

Green tides have occurred every year from 2007 to 2014 in the Yellow Sea. Ulva prolifera(Müller) J. Agardh has been identified as the bloom-forming alga,co-occurring with U. intestinalis. We observed distinct strategies for both algal species during green tides. U. prolifera exhibited a high abundance initially and then decreased dramatically,while U. intestinalis persisted throughout. The antioxidant system responses of these two macroalgae were compared in the late phase of a green tide(in-situ) and after laboratory acclimation. Lipid peroxidation and antioxidant system responses differed significantly between the two. Malondialdehyde and hydrogen peroxide contents increased significantly in-situ in U. prolifera,but not in U. intestinalis. In U. prolifera,we observed a significant decrease in total antioxidant ability(T-AOC),antioxidant enzymes(SOD and Apx),and non-enzyme antioxidants(GSH and As A) in-situ. U. intestinalis showed the same pattern of T-AOC and SOD,but its Gpx,Apx,and GSH responses did not differ significantly. The results suggest that U. prolifera was more susceptible than U. intestinalis to the harsh environmental changes during the late phase of a Yellow Sea green tide. The boom and bust strategy exhibited by U. prolifera and the persistence of U. intestinalis can be explained by differences in enzyme activity and antioxidant systems.     

The distribution of green algal micro-propagules and macroalgae at the early stage of green tide in the coastal area of South Jiangsu Province in 2014

An integrated investigation was carried out during late April to late May of 2014 in the coastal area of South Jiangsu Province to understand the early development of green tide in the Yellow Sea and discover the temporal and spatial distribution of green algal micro-propagules and macroalgae at the early stage of green tide. The results showed that green algal micro-propagules distributed in all stations from late April to late May, and micro-propagules of high density( 300 ind. L~(-1)) concentrated in the adjacent Porphyra aquaculture area. Floating macroalgae were initially observed in the northern survey areas, and increased gradually. The dominant species was U. prolifera in the floating macroalgae, ranging between 90% and 100%. The seawater surface temperature and salinity in the above mentioned time period were suitable for the growth of macroalgae. This work can help government to strengthen management to reduce the blooms of macroalgae in the coastal area of South Jiangsu Province and also facilitate the decision-making for managers at the early stage of green tide.     

Tidal current observation in the southern Yellow Sea in the summers of 2001 and 2003

Direct current observations in the Yellow Sea interior are very scarce due to intense fishing and trawling activities. Most previous studies on tides in the area were based on coastal measurements or satellite altimeter sea levels and have not been rigorously compared with direct measurements. In this paper, tidal currents are studied with current profiles from three bottom-moored Sontek Acoustic Doppler Profilers （ADPs） deployed in the southern Yellow Sea in summer of 2001 and 2003. The measured current series were dominated by tidal currents. Maximum velocities are between 40-80 cm/s at the mooring stations. M2 current is the most dominant primary tidal constituent, while MS4 and M4 are the most significant shallow water tides with much smaller amplitudes than the primary tides.     

A LAYERED NUMERICAL MODEL FOR SIMULATING THE GENERATION AND PROPAGATION OF INTERNAL TIDES OVER CONTINENTAL SLOPE Ⅲ. NUMERICAL EXPERIMENTS AND SIMULATION

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Comparison of environmental responding strategies between Ulva prolifera and Sargassum horneri: an in-situ study during the co-occurrence of green tides and golden tides in the Yellow Sea,China in 2017

Journal of Oceanology and Limnology - Large-scale green tides in the Yellow Sea occurred for 13 consecutive years since 2007. The unusual co-occurrence of green tides and golden tides occurred in...     

Phylogenetic analyses of four species of Ulva and Monostroma grevillei using ITS, rbc L and 18S rDNA sequence data

Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-1 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulva prolifera (or U. pertusa) gene sequences from Qingdao and Nan’ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U. flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.     

A layered numerical model for simulating the generation and propagation of internal tides over continental slope III. Numerical experiments and simulation

The layered model in Part I was used to simulate the internal tide in a stratified, two layer, and rectangular sea area with step-like topography. The internal tide current velocities of the upper and lower layers and the interfacial elevations were computed and the effect of the upper layer water depth and density difference were studied. Numerical experiments verified that the model can correctly simulate internal tides. The model was also applied to the northwestern part of the South China Sea to simulate the internal tides there with real topography. The distributions of internal tide amplitude in interfaces were delineated. Contribution No. 3590 from the Institute of Oceanology, Chinese Academy of Sciences. Project 49676275 supported by NSFC and also supported by National Special Research Program 97-926-05-02.     

Flocculation of Ulva microscopic propagules using modified clay: a mesocosm experiment

Microscopic propagules of Ulva species(UMPs) spread on Subei Shoal are believed to play a significant role in the formation of the Yellow Sea green tide.Previous laboratory and ship-based studies indicated that modified clay(MC) can effectively remove UMPs from the water column and restrict their germination.To evaluate the effectiveness of this method under natural conditions,a mesocosm experiment was conducted on the Zhugensha sandbank on Subei Shoal.The results show that the suspended particles on the sandbank would partially disturb the flocculation process between the MC and UMPs.However,the MC at a proper dose could effectively remove UMPs from the water column and decrease the adhesion and germination of UMPs on the nylon ropes that comprised the mariculture rafts by75%.This method was proven a potential strategy to restrict the initial biomass accumulation of green algae on Subei Shoal.However,field trials in larger time-space scales are still needed to assess the efficiency of this method when used in the open sea.     

Mitochondrial DNA control region diversity and population structure of Pacific herring (Clupea pallasii) in the Yellow Sea and the Sea of Japan

To investigate the genetic variation and population structure of Pacific herring in the Yellow Sea and the genetic differentiation between the Yellow Sea and the Sea of Japan,fragments of 479-bp mitochondrial DNA control region were sequenced for 110 individuals collected from three different periods in the Yellow Sea and one locality in the Sea of Japan.High haplotype diversity and moderate nucleotide diversity were observed in Pacific herring.AMOVA and exact test of population differentiation showed no significant genetic differentiations among the three populations of the Yellow Sea and suggested the populations can be treated as a single panmictic stock in the Yellow Sea.However,a large and significant genetic differentiation(WST50.11;P50.00) was detected between the populations in the Yellow Sea and the Sea of Japan.The high sea water temperature in the Tsushima Strait was thought a barrier to block the gene exchange between populations of the two sea areas.The neutrality tests and mismatch distribution indicated recent population expansion in Pacific herring.     

The green tide in Yingkou,China in summer 2021 was caused by a subtropical alga—Ulva meridionalis (Ulvophyceae,Chlorophyta)

Journal of Oceanology and Limnology - The large-scale green tide caused by Ulva has occurred successively in the Yellow Sea since 2007, and new events of green tide also continued to appear in...     

An investigation of the properties of low-frequency internal waves in the northeastern China Seas

A simple and practical method for separating low-frequency internal waves from low-frequency barotropic waves was employed to analyze the observation data. Analysis of some data gathered in the northestern China Seas revealed strong semidiurnal internal tides and near-inertial internal waves at the stations in the East China Sea and near-inertial waves but no semidiurnal internal tides at the station in the centre of the Yellow Sea. The geographic properties of low-frequency internal waves in the region are discussed primarily on the basis of the mechanism of internal tide generation on the continental shelfbreak, and the mechanisms of local generation and global generation. Project supported by the National Natural Science Foundation of China.     

Identification of two Skeletonema costatum-like diatoms by fluorescence in situ hybridization

A harmful algae bloom (HAB) is a dense aggregation of algae in a marine or aquatic environment that can result in significant environmental problems. To forecast the occurrence of HAB, development of a rapid and precise detection method is urgently required. In this study, two Skeletonema costatum-like diatoms (SK-1 and SK-2), were identified morphologically under a light microscope, and detected using fluorescent in situ hybridization (FISH). Strain SK-1 was isolated from a frequently HAB affected area of the East China Sea, and strain SK-2 from an aquatic farm in Qingdao, China. Fluorescent DNA probes were designed that were complementary to the ITS sequence (including 5.8S rDNA) of strain SK-1. After hybridization, strong green fluorescence was observed in cells of strain SK-1 under an epifluorescence microscope; however, no such fluorescence was observed with strain SK-2, which indicates that probes hybridized only the DNA of the target strain, SK-1, in species-specific manner, and that the two strains do not belong to a same species. This finding was confirmed by ITS sequence analysis. The FISH technique used in this study was sensitive, simple, and rapid, and is a promising tool for detecting target HAB species in natural environments.