Relationship between toxic and harmful microalgae and environmental factors in typical mariculture areas of East China Sea

Toxic and harmful algal blooms are usually more frequent in mariculture areas due to the abundant trophic conditions. To investigate the relationship between toxic and harmful microalgae and environmental factors, we set up 12 stations near three mariculture regions (Gouqi Island, Sandu Bay, and Dongshan Bay) in the East China Sea. We collected samples from all four seasons starting from May 2020 to March 2021. We identified 199 species belonging to 70 genera, of which 38 species were toxic and harmful, including 24 species of Dinophyceae, 13 species of Bacillariophyceae, and 1 species of Raphidophyceae. The species composition of toxic and harmful microalgae showed a predominance of diatoms in the summer (August), and dinoflagellates in the spring (May), autumn (November), and winter (March). The cell densities of toxic and harmful microalgae were higher in summer (with an average value of 15.34×10³ cells/L) than in other seasons, 3.53×10³ cells/L in spring, 1.82×10³ cells/L in winter, and 1.0×10³ cells/L in autumn. Pseudonitzschia pungens, Prorocentrum minimum, Paralia sulcata, and Prorocentrum micans were the dominant species and were available at all 12 stations in the three mariculture areas. We selected 10 toxic and harmful microalgal species with frequency >6 at the survey stations for the redundancy analysis (RDA), and the results show that NO ^?₃ , water temperature (WT), pH, DO, and NO ^?₂ were the main factors on distribution of toxic and harmful microalgae. We concluded that the rich nutrient conditions in the East China Sea mariculture areas increased the potential for the risk of toxic and harmful microalgal bloom outbreaks.

     

Screening for lipophilic marine toxins and their potential producers in coastal waters of Weihai in autumn, 2020

Lipophilic marine toxins (LMTs) produced by some microalgae in the sea could accumulate in shellfish and pose potential threats to the health of seafood consumers. Phytoplankton and shellfish samples were collected from coastal waters of Weihai in Shandong Peninsula, China in autumn, 2020, and screened for lipophilic marine toxins and their potential producers using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and high throughput sequencing of partial DNA (V4 region of the 18S rRNA gene) extracted from phytoplankton. Pectenotoxin-2 (PTX2), trace amounts of azaspiracid (AZA1 or AZA40), and 13-desmethyl spirolide C (13-DesMe-C) were detected in phytoplankton samples, while PTX2 and gymnodimine (GYM) were detected in shellfish samples. The toxin content in shellfish samples was much lower than the regulatory limit or values reported previously. Results suggest that lipophilic marine toxins should have low risk in coastal waters of Weihai in autumn. Based on the data of high throughput sequencing, the OTUs were assigned to 5 identified species of Alexandrium, including A. ostenfeldii capable of producing 13-DesMe-C and GYM. Two OTUs were found closely related to the toxic species in genus Dinophysis, but it is impossible to assign them to any identified species due to the low resolving power of the V4 region for Dinophysis. The OTUs could not be assigned to any identified species in the genus Azadinium, suggesting the existence of unidentified species in this region.

     

Revealing an Invasion Risk of Fish Species in Qingdao Underwater World by Environmental DNA Metabarcoding

Environmental DNA(eDNA)metabarcoding has emerged as a potentially powerful tool to monitor invasive fish species.As an alternative(or complementary)tool for biodiversity monitoring,e DNA metabarcoding had been used to detect species in aquariums,which represents an important transit avenue for introducing non-indigenous species with high population densities.In this study,eDNA metabarcoding as well as morphological characterization were used to reveal the diversity of non-indigenous species in a large aquarium at Qingdao Underwater World.Environmental DNA metabarcoding of 14 water samples at five locations from the Big Water Tank detected 24 non-indigenous species and four putative non-indigenous operational taxonomic units(OTUs).In contrast,only 20 non-indigenous species were observed by morphological characterization.Some species undetected by morphological characterization,such as Oreochromis niloticus(Linnaeus,1758),are highly adaptable to various environments and/or have invaded preferred regions where they threaten native aquatic species.eDNA metabarcoding also detected seven local fishes that were not identified by morphological characterization.However,analysis of OTU diversity among stations and sample replications revealed that eDNA varied within and/or between stations.Increasing sampling effort as well as negative controls are required to increase the detection rate of species and to eliminate false-positive OTUs.     

Will harmful dinoflagellate Karenia mikimotoi grow phagotrophically?

We studied the phagotrophic ability of dinoflagellate strain Karenia mikimotoi KM-Lü (isolated from the South China Sea), using fluorescent microspheres, bacteria isolated from the culture of K. mikimotoi and a marine microalgae Isochrysis galbana. We found that K. mikimotoi cultured under conditions of high light intensity could ingest fluorescent microspheres (diameters 0.5 and 2.0 μm) and fluorescence-labeled bacteria and microalgae. Under a low light intensity, however, only fluorescent microspheres (diameter 0.5 μm) and fluorescence-labeled microalgae were ingested. K. mikimotoi showed better growth by ingesting living marine bacteria or microalgae I. galbana than the controls, either in nutrient-depleted or nutrient-replete conditions. In nutrient-depleted conditions, the growth of K. mikimotoi was more significant with I. galbana as the prey item. In conclusion, the harmful dinoflagellate K. mikimotoi from the South China Sea has apparent phagotrophic ability, and some marine bacteria and microalgae may promote the growth of K. mikimotoi.     

Distributions of inorganic nutrients in the bohai sea of china

1　Introduction　TheBohaiSeaislocatedinthenorthernChinawithlongitudesofbetween 117°38′Eand 12 2°31′Eandlat itudesofbetween 37°0 8′Nand 4 1°0 2′N .Itisashal lowseawithanaveragewaterdepthof 18m (LiuandZhang ,2 0 0 0 ) .Severalbigrivers ,suchastheLiaoheRiver,theHaiheRiverandtheYellowRiver ,findtheirwaysintotheBohaiSeaandtransportlargeamountofnutrientsandsuspendedmattersfromthecontinentintothesea (Zhangetal.,1994 ;Zhang ,1996 ) .Duringthelasttwodecades ,marineenviron mentintheBohai…     

Molecular analyses of bacterioplankton communities with highly abundant Vibrio clades: a case study in Bohai Sea coastal waters

We investigated the bacterioplankton abundance,community composition and the associated Vibrio clades of natural seawater in Bohai Sea coastal waters.Seawater samples(10 L in triplicate)were collected at 0.5,3,and 5 m depths near the coastal aquaculture zone of the Bohai Sea on May 12,2016.Real-time PCR and 16 S rRNA gene amplicon high-throughput sequencing methods were employed by which 485 operational taxonomic units(OTUs)at a 97% sequence similarity level were generated.Gammaproteobacteria,Alphaproteobacteria,and Bacteroidetes were the most abundant groups,accounting for 49.5%,23.5%,and 18.9% ofthe total assemblage,respectively.Obvious variations in Pseudoalteromonas,Vibrio,and Octadecabacter,which were the most abundant genera,could be observed among different samples.Notably,the results of Vibrio-specific real-time PCR indicated that Vibrio had extremely high 16 S rRNA gene copy numbers.The 16 S rRNA gene sequencing results across all the samples also indicated that they occupied a large proportion of the total assemblage.Both the alpha diversity and major bacterioplankton group Pseudoalteromonas had significant correlations with the concentration of PO43-.Overall,studies on bacterioplankton communities with highly abundant Vibrio clades can provide interesting insight into the microbial function and health assessment of the Bohai Sea coastal ecosystem.     

Characteristics of phytoplankton community structure during and after a bloom of the dinoflagellate Scrippsiella trochoidea by HPLC pigment analysis

A bloom of the dinoflagellate Scrippsiella trochoidea was detected for the first time in inner Tolo Harbor, Hong Kong in 2 000. Water samples were collected at eight stations along a transect passing through a red tide patch for microscopic analysis of phytoplankton composition and high-performance liquid chromatography (HPLC) analysis of phytoplankton pigments. During the bloom, the density of dinoflagellates was 1.1×10⁶ cells L⁻¹ within the patch and 8.6×10⁵ cells L⁻¹ outside the patch where the phytoplankton community was dominated by diatoms. After the bloom the S. trochoidea began to decrease in density and was replaced by diatoms as the dominating bloom-causing organisms at all stations, and the density of dinoflagellates at most stations was less than 1.0 × 10⁶ cells L⁻¹. The status of S. trochoidea as the causative species of the bloom was indicated by the presence of peridinin, the marker pigment for dinoflagellates. The shift from dinoflagellates to diatoms was marked by the decline of peridinin and the prevalence of fucoxanthin. Phytoplankton pigment markers also revealed the presence of other minor phytoplankton assemblages such as cryptomonads and blue-green algal.     

Effects of co-existing microalgae and grazers on the production of hemolytic toxins in Karenia mikimotoi

Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought to cause mortality in fishes and invertebrates. We evaluated the composition of the hemolytic toxin produced by K. mikimotoi cultured in the laboratory using thin-layer chromatography.In addition, we evaluated the effect of co-occuring algae (Prorocentrum donghaiense and Alexandrium tamarense) and the cladoceran grazer Moina mongolica on hemolytic toxin production in K. mikimotoi. The hemolytic toxins from K. mikimotoi were a mixture of 2 liposaccharides and 1 lipid. Waterborne clues from P. donghaiense and A. tamarense inhibited the growth of K. mikimotoi but increased the production of hemolytic toxins. Conversely, K. mikimotoi strongly inhibited the growth of caged P. donghaiense and A. tamarense. In addition, the ingestion of K. mikimotoi by M. mongolica induced the production of hemolytic toxins in K. mikimotoi. Taken together, our results suggest that the presence of other microalgae and grazers may be as important as environmental factors for controlling the production of hemolytic substances. K. mikimotoi secreted allelochemicals other than unstable fatty acids with hemolytic activity. The production of hemolytic toxins in dinoflagellates was not only dependent on resource availability, but also on the risk of predation. Hemolytic toxins likely play an important role as chemical deterrents secreted by K. mikimotoi.     

Application of an integrated methodology for eutrophication assessment: a case study in the Bohai Sea

An integrated methodology for eutrophication assessment,which integrates both water quality indicators(causative factors) and ecological response indicators(effect factors),is described.It is then applied to rank the eutrophication status of the years 2007 and 2008 in the southwest Bohai Sea.The assessment model identified that north Bohai Bay and west Laizhou Bay were the two areas with the most serious eutrophication problems in the southwest Bohai Sea.In addition,compared to that in the west Laizhou Bay,the eutrophication conditions in the north Bohai Bay was more serious in both years.Eutrophication problems such as harmful algal blooms(HABs) and low dissolved oxygen(DO) events in north Bohai Bay were frequent.The integrated method outmatched the currently used Chinese nutrient index method by definitely identifying areas with the most serious eutrophication problems,while the nutrient index method gave ambiguous results between the two years.Inclusion of both causative factors and effect factors,combining concentration,spatial coverage and frequency of indicators,as well as use of multi-season monitoring datasets in the methodology result in a more accurate,representative and useful assessment.     

Biodiversity and distribution of benthic dinoflagellates in tropical Zhongsha Islands,South China Sea

Journal of Oceanology and Limnology - Benthic dinoflagellates have attracted increasing attention in recent years because of their toxicity and ability to form extensive harmful algal blooms....     

DISTRIBUTION AND POPULATION DYNAMICS OF PARACALANUS PARVUS,PRACALANUS CRASSIROSTRIS,AND ACARTIA BIFILOSA（COPEPODA,CALANOIDA）IN THE BOHAI SEA

Paracalanus parvus,Paracalanus crassirostris,and Acartia bifilosa are dominant and widely distributed in the Bohai Sea,and comprise an important part of zooplankton in terms of biomass as well as production rate.In order to understand their seasonal distribution and population dynamics,their stage-specific abundance in different months of the year were analyzed based on the never analyzed yet samples collected in 1959.The three species showed clear and remarkable seasonal variation in abundance,which maximized in spring and summer,when they formed high biomass patches or concentrations in the nearshore area.For Paracalanus parvus,two peaks were observed in the annual circle,one in June and the other in September,For Paracalanus crassirostris,one peak occurred in summer and a small one in December.The seasonal pattern of Acartia bifilosa was different in different regions.In Bohai Bay it had a two-peak pattern,with the first large peak occurring in May and the second one in October.In Laizhou Bay,a winter peak in December and January could be observed besides the spring one.The number of generations during the reproductive season for the three species was estimated based on the annual cycle in abundance and ambient temperature.     

Changes in nitrogen and phosphorus and their effects on phytoplankton in the Bohai Sea

Systematic studies of the changes in dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP) and their effects on phytoplankton over the last 30 years in the Bohai Sea are presented. The amount of sewage disposal, use of fertilizer and the Huanghe River runoff were found to have a significant influence on the DIN or DIP concentrations in the Bohai Sea over the last 30 years. Moreover, the changes in DIN and DIP resulted in changes in the limiting nutrients of phytoplankton in the Bohai Sea from nitrogen in the early 1980s to nitrogen-phosphorus in the late 1980s, and then to phosphorus after the 1990s. In addition, changes in nitrogen and phosphorus had a significant effect on the phytoplankton community structure. The half saturation constant (K _s) was used to evaluate the effect of nutrients on the phytoplankton community structure in the Bohai Sea over the last 30 years. Cell abundance percentages of dominant phytoplankton species with high K _s values for phosphorus and low K _s values for nitrogen have decreased since the 1980s, while those of dominant phytoplankton species with low K _s values for phosphorus and high K _s values for nitrogen increased during this period.     

Distribution patterns of phytoplankton in the Changjiang River estuary and adjacent waters in spring 2009

The Changjiang River estuary and adjacent waters are one of the most notable regions for red tides/harmful algal blooms in China's coastal waters.In this study,phytoplankton samples were collected and analyzed during the outbreak stage of red tides in May 2009.It was found that dinoflagellates,Prorocentrum donghaiense and Karenia mikimotoi,and diatoms,Skeletonema spp.and Paralia sulcata,were the major taxa dominating the phytoplankton community.Cluster analysis,non-metric multidimensional scaling(NMDS) and analysis of similarities(ANOSIM) was conducted on a data matrix including taxa composition and cell abundance of the phytoplankton samples.The analyses categorized the samples into three groups at a similarity level of 30%.Group Ⅰ was characterized by estuarine diatoms and distributed mainly in the highly turbid estuarine region.Group Ⅱ,which was dominated by the diatom Skeletonema spp.and represented the red tide of Skeletonema spp.,was situated around Group Ⅰ in the sea area west of 122°50'E.Group Ⅲ was characterized by a high proportion of dinoflagellates and was found further offshore compared with Groups Ⅰ and Ⅱ.Group Ⅲ was further divided into two subgroups(Ⅲ-S1 and Ⅲ-S2) at a similarity level of 40%.Group Ⅲ-S1 was characterized by the presence of the benthic diatom P.sulcata,representing phytoplankton samples collected either from the bottom or from the sea area affected by upwelling.GroupⅢ-S2 was dominated by dinoflagellates and represented red tides formed by P.donghaiense and K.mikimotoi.A gradual change of red-tide causative species was observed from the estuary to the offshore sea area,from diatoms to armored dinoflagellates and then unarmored dinoflagellates.Environmental factors associated with each group,and thus affecting the distribution of phytoplankton and red tides,are discussed.     

Laboratory study on the ecological impact of sophorolipid used for harmful algae elimination

We studied the role of sophorolipid in inhibiting harmful algae bloom (HAB). Different sophorolipid concentrations were tested on marine microalgae, zooplankton, fish, and bivalve (Mytilus edulis) in laboratory. The result shows that sophorolipid could inhibit the growth of algal species selectively. Among three algae species selected, Platymonas helgolandica var. tsingtaoensis was promoted with increasing sophorolipid concentration; Isochrysis galbana was inhibited seven days later in sophorolipid concentration below 40 mg/L; and Nitzschia closterium f. minutissima was inhibited obviously in only a high sophorolipid concentration over 20 mg/L. Therefore, sophorolipid in a low concentration at <20 mg/L could remove certain harmful algae species effectively without harming other non-harmful microalgae. For other animals, sophorolipid could inhibit the growth of ciliate Strombidium sp. by 50% at 20 mg/L sophorolipid concentration after 96 h. The concentration in 96-h LC₅₀ for Calanus sinicus, Neomysis awatschensis, Lateolabrax japonicus, and Paralichthys olivaceus was 15, 150, 60, and 110 mg/L, respectively. The 24 h LC₅₀ value for Artemia salina was 600 mg/L. The relative clearance rate of mussel Mytilus edulis decreased to 80%, 40%, and 20% of the control group after being exposed to 20, 50, and 100 mg/L sophorolipid for 24 h. Therefore, the toxicity for mitigation of harmful algae bloom at previously recommended concentration of 5–20 mg/L sophorolipid is low for most tested organisms in this reaserch.     

Increased diversity and environmental threat of harmful algal blooms in the Southern Yellow Sea,China

Journal of Oceanology and Limnology - Harmful algal blooms (HABs) in the Southern Yellow Sea (SYS) have shown a trend of increasing diversity and detrimental effects. While the Bohai Sea, East...     

Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

Overthelastseveraldecades,harmfulalgalblooms(HABs)havebecomeaseriousenvironmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.,) were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.     

Taxonomic distinctness of macrofauna as an ecological indicator in Laizhou Bay and adjacent waters

In this paper, we assessed the ecological and biodiversity status in the Bohai Sea through a quantitative survey on mac-rofaunal community at 25 stations in Laizhou Bay and adjacent waters in the autumn of 2006.We tested the robustness and effectiveness of taxonomic distinctness as an ecological indictor by analyzing its correlation with species richness and natural environmental variables and by analyzing other ecological indicators (Shannon-Wiener H' and W statistics from Abundance Biomass Comparison curve).Results so obtained indicated that the benthic environment of the study waters in general is not under major impact of anthropogenic disturbance, but some stations in Laizhou Bay and along the coast of the Shandong Peninsula and even in the central Bohai Sea might be moderately disturbed and showed signs of ecological degradation.The taxonomic distinctness measures △+ and Λ+ were independent of sampling effort and natural environment factors and were compliant to other ecological indicators.Further application of the taxonomic distinctness indicator to assess marine biodiversity and ecosystem health on a larger regional scale with historical data seems promising.     

The observed currents in summer in the Bohai Sea

A harmonic method was used to analyze the tidal currents observed in summer at 11 stations made from 1996 to 2001 in the Bohai Sea, China. Data was compared among different instruments and intervals. Elliptic elements were calculated based on harmonic constants, of which vertical distributions of the maximum speed and rotation direction were discussed for understanding the characteristics of diurnal and semi-diurnal tidal current components. The results indicate that the maximum speed of M2 tidal current component is much larger than that of K1; the rotation direction of M2 tidal current constituent is clockwise in the central part of the Bohai Sea and in the Laizhou Bay, but anticlockwise in the Liaodong Bay and Bohai Bay. For K1 tidal current constituent, it is clockwise in the central Bohai Sea but anti-clockwise in the Laizhou Bay and Liaodong Bay. The tidal currents in most stations in the Bohai Sea were regular semidiurnal except for those in the central Bohai Sea, being irregular semidiurnal.     

Physicochemical conditions in affecting the distribution of spring phytoplankton community

To better understand the physicochemical conditions in af fecting regional distribution of phytoplankton community, one research cruise was carried out in the Bohai Sea and Yellow Sea during 3 rd and 23 th May, 2010. The phytoplankton community, including Bacillariophyta(105 taxa), Pyrrophyta(54 taxa), Chrysophyta(1 taxon) and Chlorophyta(2 taxa), had been identified and clearly described from six ecological provinces. And, the six ecological provinces were partitioned based on the top twenty dominant species related with notable physicochemical parameters. In general, the regional distributions of phytoplankton ecological provinces were predominantly influenced by the physicochemical properties induced by the variable water masses and circulations. The predominant diatoms in most of water samples showed well adaptability in turbulent and eutrophic conditions. However, several species of dinoflagellates e.g., Protoperidinium conicum, Protoperidinium triestinum, Protoperidinium sp. and Gymnodinium lohmanni preferred warmer, saltier and nutrient-poor environment. Moreover, the dinoflagellates with high frequency in the Yellow Sea might be transported from the Yellow Sea Warm Current. The horizontal distribution of phytoplankton was depicted by diatoms and controlled by phosphate concentration, while the vertical distribution was mainly supported by light and nutrients availability in the subsurface and bottom layers, respectively.