Xuelei Zhang

Effect of temperature, salinity and irradiance on growth and photosynthesis of Ulva prolifera

Intensive Pyropia aquaculture in the coast of southwestern Yellow Sea and its subsequent waste, including disposed Ulva prolifera, was speculated to be one of the major sources for the large-scale green tide proceeding in the Yellow Sea since 2007. It was, however, unclear how the detached U. prolifera responded and resumed growing after they detached from its original habitat. In this study, we investigated the growth and photosynthetic response of the detached U. prolifera to various temperature, salinity and irradiance in the laboratory. The photosynthetic rate of the detached U. prolifera was significantly higher at moderate temperature levels (14–27°C) and high salinity (26–32), with optimum at 23°C and 32. Both low (<14°C) and highest temperature (40°C), as well as low salinity (8) had adverse effects on the photosynthesis. Compared with the other Ulva species, U. prolifera showed higher saturated irradiance and no significant photoinhibition at high irradiance, indicating the great tolerance of U. prolifera to the high irradiance. The dense branch and complex structure of floating mats could help protect the thalli and reduce photoinhibition in field. Furthermore, temperature exerted a stronger influence on the growth rate of the detached U. prolifera compared to salinity. Overall, the high growth rate of this detached U. prolifera (10.6%–16.7% d–1) at a wide range of temperature (5–32°C) and salinity (14–32) implied its blooming tendency with fluctuated salinity and temperature during floating. The environmental parameters in the southwestern Yellow Sea at the beginning of green tide were coincident with the optimal conditions for the detached U. prolifera.

Community structure and controlled factor of attached green algae on the Porphyra yezoensis aquaculture rafts in the Subei Shoal, China

This study analyzes the community structure, the quantity changes of the algae and the effect of important environmental factors and estimates the total biomass of the attached green algae in the survey areas. The study uses data from surveys of the attached green algae on the Porphyra yezoensis aquaculture rafts and data regarding the environmental factors from October 2010 to April 2011 in the Subei Shoal. The attached green algae on the rafts included Ulva prolifera, Capsosiphon groenlandicus, U. linza, U. intestinalis, U. clathrata, and U. compressa. The biomass changes of the attached green algae exhibited an inverted parabola: the biomass was the highest (14 898 t) in April, and was the second highest (2 034 t) in November; it was lowest in February (only 729 t) and increased sharply from March to April. The species diversity differed significantly among the seasons. In September and October, when the P. yezoensis aquaculture rafts were initially set up, the attached green algae had a high biodiversity, while from December to the next February, a variety of green algae species coexisted on the rafts, although the biomass was low, and from March to April, as the biomass increased sharply, the species diversity dropped to the minimum. During this time, C. groenlandicus was apparently dominant with the maximum biomass proportion up to 80%, while the U. prolifera proportion increased exponentially to 20% to 40%. The water temperature had a direct regulating effect on the biomass and the species succession of the attached green algae. The estimation of the community dynamics and the biomass of the green algae provided the evidence needed to track the origin of the large-scale green tide in the southern Yellow Sea.

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.

The current situations of green macroalgae and micro-propagules in Pyropia aquaculture of the Subei Shoal in spring of 2013

To discover the distribution of green algal micro-propagules in the Subei Shoal and clarify the growth of green macroalgae attached on Pyropia aquaculture rafts, an integrated investigation in Pyropia aquaculture area and one cruise in the coastal area of the Subei Shoal were carried out from March to May in 2013. The results showed that green algal micro-propagules were discovered in seawater and sediment during March to May. The average quantity of micro-propagules was 267 ind./L in surface seawater and 43 ind./g in sediment. The biomass of attached green macroalgae increased in Pyropia aquaculture from March to May. Three species, including Ulva prolifera, Ulva linza and Blidingia sp. were found in Pyropia aquaculture rafts. The dominant specie was Blidingia sp. and the second was U. prolifera in spring. This study indicated that the micro-propagules and macroalgae were existed in the coastal area of the Subei Shoal at the early stage of green tide. This was the key point to the governance of green tide in China.

Comparative study of the hemolytic and cytotoxic activities of nematocyst venoms from the jellyfish Cyanea nozakii Kishinouye and Nemopilema nomurai Kishinouye

Two species of jellyfish, Cyanea nozakii Kishinouye and Nemopilema nomurai Kishinouye, have occurred offcoastal areas of the northeastern China Sea, Yellow Sea, and Bohai Sea in recent years. They influence marine ecosystem safety and fishery production, and also pose a risk to human health. The current study examined the hemolytic and cytotoxic activities of crude venoms extracted from the nematocysts of C. nozakii and N. nomurai. The results showed that there were more nematocysts on tentacles from C. nozakii than on tentacles of the same length from N. nomurai. The protein concentration per nematocyst extracted from N. nomurai was higher than that from C. nozakii. Both nematocyst venoms showed dose- and timedependent hemolytic activity on erythrocytes from chicken, pigeon, and sheep, with sheep erythrocytes being the most sensitive, with EC50 values of 69.69 and 63.62 μg/mL over a 30-min exposure with N. nomurai and C. nozakii nematocyst venoms, respectively. A cytotoxic assay of both jellyfish venoms on A431 human epidermal carcinoma cells resulted in IC50 values of 68.6 and 40.9 μg/mL after 24-h incubation, respectively, with venom from C. nozakii showing stronger cytotoxic activity than that from N. nomurai. The results of current study indicate that nematocyst venom from C. nozakii had stronger hemolytic and cytotoxic activities than that from N. nomurai and, thus, C. nozakii might be more harmful to the health of humans and other species than are N. nomurai when they appear in coastal waters.

Ethanol Extract from Ulva prolifera Prevents High-Fat Diet-Induced Insulin Resistance, Oxidative Stress, and Inflammation Response in Mice

Ulva prolifera is the major causative species in the green tide, a serious marine ecological disaster, which bloomed in the Yellow Sea and the Bohai Sea of China. However, it is also a popular edible seaweed and its extracts exerts anti-inflammatory and antioxidant effects. The present study investigated the effects of ethanol extract of U. prolifera (EUP) on insulin sensitivity, inflammatory response, and oxidative stress in high-fat-diet- (HFD-) treated mice. HFD-treated mice obtained drinking water containing 2% or 5% EUP. The results showed that EUP supplementation significantly prevented HFD-induced weight gain of liver and fat. EUP supplementation also improved glucose tolerance and insulin resistance in HFD-treated mice. Moreover, EUP supplementation prevented the increased expression of genes involved in triglyceride synthesis and proinflammatory genes and the decreased expression of genes involved in fatty acid oxidation in liver of HFD-treated mice. Furthermore, EUP supplementation decreased reactive oxygen species content, while increasing glutathione content and glutathione peroxidase activity in HFD-treated mice. In conclusion, our results showed that EUP improved insulin resistance and had antilipid accumulation and anti-inflammatory and antioxidative effects on HFD-treated mice. We suggested that U. prolifera extracts may be regarded as potential candidate for the prevention of nonalcoholic fatty liver disease.

Source of propagules of the fouling green macroalgae in the Subei Shoal, China

Since 2007, large-scale green tides dominated by Ulva prolifera consecutively bloomed in the Yellow Sea and caused great economic losses. The fouling U. prolifera on the Pyropia yezoensis aquaculture rafts in the Subei Shoal was regarded as the major source of the floating biomass. However, it was still unclear about the seed source of fouling green macroalgae attached on the rafts. In this study, the field surveys and the indoor experiments were conducted to reveal the source of propagules of the fouling green macroalgae on the rafts and to study the anti-fouling material for P. yezoensis aquaculture rafts which could possibly be a feasible strategy to control the green tides in the Yellow Sea. The results showed that (1) micro-propagules of several green macroalgal species, including U. prolifera, U. linza, U. compressa, U. flexuosa, and Blidingia sp. coexisted in the waters and sediments in the Subei Shoal and their proportion remarkably changed over time; (2) the bamboo poles with peeling treatment could significantly reduce the amount of U. prolifera micro-propagules attached. This study confirmed that the micro-propagules distributed in the Subei Shoal area were the precursors of the green tides, and provided a feasible method to control the Yellow Sea large-scale green tides at the beginning.

Effect of CO2 on growth and toxicity of Alexandrium tamarense from the East China Sea, a major producer of paralytic shellfish toxins

In recent decades, the frequency and intensity of harmful algal blooms (HABs), as well as a profusion of toxic phytoplankton species, have significantly increased in coastal regions of China. Researchers attribute this to environmental changes such as rising atmospheric CO2 levels. Such addition of carbon into the ocean ecosystem can lead to increased growth, enhanced metabolism, and altered toxicity of toxic phytoplankton communities resulting in serious human health concerns. In this study, the effects of elevated partial pressure of CO2 (pCO2) on the growth and toxicity of a strain of Alexandrium tamarense (ATDH) widespread in the East and South China Seas were investigated. Results of these studies showed a higher specific growth rate (0.31±0.05day-1) when exposed to 1000μatm CO2, (experimental), with a corresponding density of (2.02±0.19)×107cellsL-1, that was significantly larger than cells under 395μatm CO2(control). These data also revealed that elevated pCO2 primarily affected the photosynthetic properties of cells in the exponential growth phase. Interestingly, measurement of the total toxin content per cell was reduced by half under elevated CO2 conditions. The following individual toxins were measured in this study: C1, C2, GTX1, GTX2, GTX3, GTX4, GTX5, STX, dcGTX2, dcGTX3, and dcSTX. Cells grown in 1000μatm CO2 showed an overall decrease in the cellular concentrations of C1, C2, GTX2, GTX3, GTX5, STX, dcGTX2, dcGTX3, and dcSTX, but an increase in GTX1 and GTX4. Total cellular toxicity per cell was measured revealing an increase of nearly 60% toxicity in the presence of elevated CO2 compared to controls. This unusual result was attributed to a significant increase in the cellular concentrations of the more toxic derivatives, GTX1 and GTX4.Taken together; these findings indicate that the A. tamarense strain ATDH isolated from the East China Sea significantly increased in growth and cellular toxicity under elevated pCO2 levels. These data may provide vital information regarding future HABs and the corresponding harmful effects as a result of increasing atmospheric CO2.

Genetic Variations of the Parasitic Dinoflagellate Hematodinium Infecting Cultured Marine Crustaceans in China.

The parasitic dinoflagellate Hematodinium infects multiple cultured marine crustaceans and has resulted in significant economic losses to their aquaculture in China. Limited molecular data implied a close relationship among Hematodinium reported in China, whereas the genetic diversity and detailed genetic variation within Hematodinium remains unclear. In order to investigate the genetic diversity and composition of the parasitic dinoflagellate in China, the sequences of the internal transcribed spacer region (ITS1 - 5.8S - ITS2) were amplified from 104 infected hosts cultured in three geographic locations. Two strains were identified, Hematodinium perezi II was the prevalent strain observed in all three host taxa throughout the survey, the other novel one was detected from the single Exopalaemon carinicauda collected from Zhejiang province. Eight ITS haplotypes within H. perezi II were identified with a similarity of 99.8% -100%. The distinct haplotype compositions and AMOVA analysis suggested a significant genetic differentiation and population structure among the three geographic populations of H. perezi II. The novel strain, genetically affiliated with H. perezi, was quite divergent from the three known genotypes (I, II and III) of H. perezi, and further investigation is needed to determine the distribution and host range of this new strain.

Artificial seed germination and seedling production of Zostera marina L. by salinity manipulation

Vast declines in Zostera marina seagrass beds demand effective methods of rehabilitation. In this study, we developed a practical method by reducing salinity to induce seed germination followed with recovering salinity to facilitate seedling production of Z. marina. The results showed that Z. marina seeds collected from natural seawater (salinity 30) were induced to germinate at reduced salinities. Percent germination (GR) was higher and mean-time-to-germinate (MTG) was shorter at lower salinities. The highest GR and shortest MTG occurred at salinity 0 (deionized freshwater). After germination in freshwater, seeds could develop into seedlings at salinities 5–30 and continue the growth. Viability or development of germinated seeds was not significantly different during the 40 d of post-germination incubation at salinities 5–15 after 1–20 d of germination in freshwater. However, during the process of translating germinated seeds from salinity 0 and 5 to salinity 30, reducing the gradients of post-germination acclimation facilitated more seeds forming seedlings in less time. On average, after 60 d of static incubation, including 20 d in freshwater for germination followed with immediate shift to salinity 5 and increasing to salinity 30 at increment of 5 every two days until cultivation at constant salinity 30, 33% of Z. marina seeds produced healthy seedlings. The results indicate that the salinity-manipulation based method of artificial germination and seedling production is practical and effective in supporting rehabilitation of Z. marina bed.