Xuexi Tang

Effects of two species of macroalgae—Ulva pertusa and Gracilaria lemaneiformis—on growth of Heterosigma akashiwo (Raphidophyceae)

The effects of fresh thalli, culture filtrate, water-soluble extract and dry powder of two species of macroalgae, Ulva pertusa (Chlorophyta) and Gracilaria lemaneiformis (Rhodophyta), on the growth of a bloom-forming microalga, Heterosigma akashiwo, were studied in co-culture under controlled laboratory conditions. Both fresh thalli and culture filtrate of U. pertusa and G. lemaneiformis, particularly in the form of fresh thalli, significantly inhibited microalgal growth; indeed, the microalga was completely killed during the course of the experiment. A clear concentration-dependent relationship was observed between the initial concentration of fresh thalli (either U. pertusa or G. lemaneiformis) and its inhibitory effect on H. akashiwo. Simultaneous nutrient assays showed that nitrate and phosphate were almost exhausted in G. lemaneiformis fresh thalli co-culture but remained well above nutrient limitation for microalgal growth in U. pertusa co-culture, in which the microalgal cells were completely killed. However, daily f/2 medium repletion would obviously alleviate the growth inhibition in G. lemaneiformis co-culture. Since the present study was carried out under controlled conditions, fluctuations in environmental factors (i.e., light, temperature, carbon limitation, bacterial presence and pH) were limited during the experiment. We thus concluded that allelopathy was the most likely explanation for microalgal growth inhibition in U. pertusa co-culture, while the combined roles of allelopathy and nutrient limitation were responsible for growth inhibition in G. lemaneiformis co-culture. Similarly, macroalgal water-soluble extracts and dry powders affected the co-cultured H. akashiwo greatly, with more obvious effects observed in water-soluble extract co-cultures. A dose-dependent relationship was also observed over the course of the experiment. It can be concluded that macroalgal thalli contain some bioactive compounds. The results of the present study suggest that U. pertusa and G. lemaneiformis, especially in the form of fresh thalli, effectively inhibit the growth of H. akashiwo and could thus be potential candidates for use in the control and mitigation of H. akashiwo blooms.

Comparative Studies on the Ecophysiological Differences of Two Green Tide Macroalgae under Controlled Laboratory Conditions

Yellow Sea green tides have occurred in coastal China almost every year from 2007 to 2011. Ulva prolifera (Müller) J. Agardh has been identified as the causative macroalgal species. U. intestinalis, however, has been observed in the bloom areas, co-occurring with U. prolifera, but it has not been found to be causative. The Yellow Sea green tide has shown consistent phases of development that match corresponding environmental changes. U. prolifera, not U. intestinalis, is dominant. Our experimental design was based on these observed phenomena, and the results of our field investigation indicated a close relationship between changes in principal environmental factors (irradiance, temperature, and salinity) and the development of each phase of the bloom. These main environmental factors were simulated to allow estimation and comparison of the physiological responses of U. prolifera and U. intestinalis. Ecophysiological differences were found between these two species. (1) More photosynthetic activity and plasticity were detected in U. prolifera. (2) U. prolifera was found to be more sensitive to dynamic environments, especially harsh and changing environmental conditions. U. intestinalis was found to be more stable, probably due to the higher stress tolerance given by its antioxidant system. (3) Markedly higher nutrient absorption activity was observed in U. prolifera. Comparisons of the ecophysiological traits of these two species in this present study may foster understanding of their natural ecological processes. Specifically, U. prolifera seemed to be more engaged with the ephemeral blooms, while U. intestinalis seemed to be directed toward persistence. This also suggests that the ecological success of U. prolifera may be inextricably linked to its higher capacity for photosynthesis, nutrient absorption, and nutrient assimilation.

Allelopathic effects of Ulva pertusa, Corallina pilulifera and Sargassum thunbergii on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense

The allelopathic effects of fresh tissue, dry powder and aqueous extracts of three macroalgae, Ulva pertusa, Corallina pilulifera and Sargassum thunbergii, on the growth of the dinoflagellates Heterosigma akashiwo and Alexandrium tamarense were evaluated using coexistence culture systems in which concentrations of the three macroalga were varied. The results of the coexistence assay showed that the growth of the two microalgae was strongly inhibited by using fresh tissue, dry powder and aqueous extracts of the three macroalga; the allelochemicals were lethal to H. akashiwo at relatively higher concentrations of the three macroalga. The macroalgae showing the most allelopathic effect on H. akashiwo and A. tamarense using fresh tissue were U. pertusa and S. thunbergii, using dry powder were S. thunbergii and U. pertusa, and using aqueous extracts were U. pertusa and C. pilulifera. We also examined the potential allelopathic effect on the two microalgae of culture filtrate of the three macroalga; culture medium filtrate initially exhibited no inhibitory effects when first added but inhibitory effects became apparent under semi-continuous addition, which suggested that continuous release of small quantities of rapidly degradable allelochemicals from the fresh macroalgal tissue were essential to effectively inhibit the growth of the two microalgae.

Effects of two polybrominated diphenyl ethers (BDE-47, BDE-209) on the swimming behavior, population growth and reproduction of the rotifer Brachionus plicatilis

Polybrominated diphenyl ethers (PBDEs) are new kinds of persistent organic pollutants (POPs) and their potential threats to the equilibrium and sustainability of marine ecosystems have raised worldwide concerns. Here, two kinds of PBDEs, tetra-BDE (BDE-47) and deca-BDE (BDE-209) were applied, and their toxic effects on the swimming behavior, population growth and reproduction of Brachionus plicatilis were investigated. The results showed that: (1) The actual concentrations of BDE-47 and -209 in the seawater phase measured by GC-MS (Gas Chromatography-Mass Spectrometer) were much lower than their nominal concentrations. (2) In accordance with the 24-hr acute tests, BDE-209 did not show any obvious swimming inhibition to rotifers, but a good correlation did exist between the swimming inhibition rate and BDE-47 concentration suggesting that BDE-47 is more toxic than BDE-209. (3) Both BDE-47 and -209 had a significant influence on the population growth and reproduction parameters of B. plicatilis including the population growth rate, the ratio of ovigerous females/non-ovigerous females (OF/NOF), the ratio of mictic females/amictic females (MF/AF), resting egg production and the mictic rate, which indicate that these parameters in B. plicatilis population were suitable for monitoring and assessing PBDEs. Our results suggest that BDE-47 and -209 are not acute lethal toxicants and may pose a low risk to marine rotifers at environmental concentrations for short-term exposure. They also accumulate differently into rotifers. Further research data are needed to understand the mechanisms responsible for the effects caused by PBDEs and to assess their risks accurately.

Ultraviolet irradiation induced oxidative stress and response of antioxidant system in an intertidal macroalgae Corallina officinalis L.

The response of the antioxidant defense system of an intertidal macroalgae Corallina officinalis L. to different dosages of UV-B irradiation was investigated. Results showed that superoxide dimutase (SOD) and peroxidase (POX) increased and then maintained at a relatively stable level when subjected to UV-B irradiation. Catalase (CAT) activity under medium dosage of UV-B irradiation (Muv) and high dosage of UV-B irradiation (Huv) treatments were significantly decreased. Ascorbate peroxidase (APX) activity first remained unaltered and then increased in Huv treatment. In addition, the assay on isozymes was carried out using non-denaturing polyacrylamide gel electrophoresis (PAGE). The activities of some SOD isoforms were altered by UV-B. Two new bands (POX V and POX VII) appeared upon exposure to all three UV-B dosages. CAT III activity was increased by low dosage of UV-B irradiation (Luv), whereas CAT III and CAT IV disappeared when the alga was exposed to Muv and Huv. Two bands of APX (APX VI and APX VII) were increased and a new band (APX X) was observed under Huv exposure. H2O2 and thiobarbituric acid reacting substance (TBARS) increased under Muv and Huv treatments. Overall, UV-B protection mechanisms are partly inducible and to a certain extent sufficient to prevent the accumulation of damage in C. officinalis.

Interaction between Chlorella vulgaris and bacteria: interference and resource competition

Research of interaction mechanism between Chlorella vulgaris and two bacterial strains (Z-QD08 and Z-QS01) were conducted under laboratory conditions. Growth rates of bacteria and C. vulgaris were tested under co-culture conditions to evaluate the effects of concentrations of C. vulgaris and bacteria on their interactions. To test whether the availability of inorganic nutrients, vitamins and trace metals affects the interactions between C. vulgaris and bacteria, experiments were performed with or without the culture medium filtrate of C. vulgaris or bacteria. The results showed that the growth of C. vulgaris was promoted at low concentrations of bacteria (5×106 cells/ml), and expressed a positive correlation with the bacteria density, whereas opposite trend was observed for treatments with high bacteria density (10×106 cells/ml and 20×106 cells/ml). The growth rate of bacteria decreased with the increasing concentrations of C. vulgaris. The growth of bacteria Z-QD08 was inhibited by C. vulgaris through interference competition, while the mechanism for interaction between bacteria Z-QS01 and C. vulgaris was resource competition. The influence of cell density on the interaction between microalgae and bacteria was also discussed. These experiments confirm some elements of published theory on interactions between heterotrophic bacteria and microalgae and suggest that heterotrophic bacteria play an important role in the development of blooms in natural waters.

Effects of Decabromodiphenyl Ether (BDE-209) on Inter-Specific Competition between Two Species of Marine Bloom-Forming Microalgae

Decabromodiphenyl ether (BDE-209), a new kind of persistent organic pollutants, was selected to investigate its influence on population growth and inter-specific competition between two species of marine bloom-forming microalgae, Heterosigma akashiwo and Karenia mikimotoi. (1)BDE-209 showed acute toxic effects on both microalgae and H. akashiwo was more sensitive from view of 96 h-EC50 and the ultrastructure variation. (2)The microalgal population growth patterns in mono-culture were density-dependent and the growth of both species in the normal co-culture was significantly depressed by competition (P<0.05) with different initial biomass ratios. BDE-209 exposure significantly changed the growth. (3) Lotka-Volterra competition model was used to simulate the interaction between the microalgae. BDE-209 exposure broke the competitive balance to make competition gradually shift in favor of H. akashiwo. Results suggested BDE-209 did have toxic effects on either microalgal growth or the inter-specific competition, which was quite different from previous reports. Further exploration of the mechanism is needed.

Allelopathic growth inhibition of Heterosigma akashiwo by the three Ulva spcieces (Ulva Pertusa, Ulva Linza, Enteromorpha intestinalis) under laboratory conditions

Allelopathic effects of several concentrations of fresh tissue, dry powder and dry tissue of three bloom-forming green macroalgae Ulva pertusa, Ulva linza and Enteromopha intestinalis on the red tide microalga Heterosigma akashiwo were evaluated in microcosms systems. The effects of macroalgae culture medium filtrate were investigated on H. akashiwo using initial or semi-continuous filtrate addition. Preliminary studies on the algicidal effects of one aqueous and four organic solvent extracts from the macroalgae on the microalga were carried out to confirm the existence of allelochemicals in the tissue of these macroalgae. The dry powder of U. pertusa was extracted with methanol, and the methanol extracts were partitioned to petroleum ether phase, ethyl acetate phase, butanol phase and distilled phase by liquid-liquid fractionation. The bioassays of the activity of every fraction were carried out on H. akashiwo. The resultant microcosms assay showed that the growth of H. akashiwo was strongly inhibited by using fresh tissues, dry powder or dry tissue of these three macroalgae, while aqueous and methanol extracts of both macroalgae had strong inhibitory effects on the growth of H. akashiwo, and the EC50 values for methanol extract of U. pertusa, U. linza or E. intestinalis were 0.016, 0.028×10−12 or 0.033×10−12, respectively. While the other three organic solvent extracts (acetone, ether and chloroform) had no apparent effect on its growth, this suggests that the allelochemicals from these three macroalgae had relatively high polarities. The activity of petroleum ether phase, ethyl acetate phase, butanol phase and distilled phase of U. pertusa methanol extract was carried out on H. akashiwo indicating that petroleum ether phase and ethyl acetate phase had stronger algicidal effect on H. akashiwo. The inhibition effect of the ethyl acetate phase was not as strong as that of petroleum ether phase, and effective concentration of petroleum ether phase was 17 mg/L for H. akashiwo. However, no significant algicidal effects were observed on the butanol phase and distilled water phase. These three macroalgae’s culture medium filtrate exhibited no apparent growth inhibitory effect on the microalga under initial filtrate addition whereas the growth of H. akashiwo was significantly inhibited under semi-continuous filtrate addition, which suggests that continuous release of small quantities of rapidly degradable allelochemicals from the fresh tissue of both macroalgae was effective in inhibiting the growth of H. akashiwo.

Using population demographic parameters to assess impacts of two polybrominated diphenyl ethers (BDE-47, BDE-209) on the rotifer Brachionus plicatilis.

Polybrominated diphenyl ethers (PBDEs) are highly persistent anthropogenic contaminants found in the environment, posing a risk to aquatic ecosystems, whereas there is a lack of data concerning their impacts on marine invertebrates. The objective of this study was to assess the relative lethal and sublethal aquatic toxicity effects of two PBDEs, BDE-47 and BDE-209 congeners, on marine zooplankton rotifer Brachionus plicatilis associated with PBDE concentrations and time of exposure. Gas chromatography-mass spectrometry (GC-MS) analyses were performed to determine actual PBDE concentrations. Rotifer population demographic parameters from life tables, including age-specific survivorship (lx), age-specific fecundity (mx), net reproductive rate (R0), intrinsic rate of increase (rm), finite rate of increase (λ), life expectancy (E0) and generation time (T), were used as measures of treatment effects. Results from this study have revealed increasingly intense negative effects on many of the rotifer demographic parameters with elevated PBDE concentrations. The population growth curves of B. plicatilis showed almost no lag phase and reached peak abundances within 11 days, while B. plicatilis exposed to BDE-209 had a lag phase of about 5 days. In addition, increased PBDE levels reduced the population abundances and peak population densities of B. plicatilis. The two PBDEs have caused the carrying capacity (K) suppressed and the negative influence turned more serious as the concentration rose. Results also revealed that the time to reach growth curve inflection point (Tp) was shortened by PBDEs to different degrees. This study not only indicated that life table demography and population growth curve studies were two important aspects used to evaluate toxicant PBDE effects, but also compared the two PBDE disruptions to the population growth and reproduction of the rotifer.

Acute and chronic toxic effects of Pb2 + on polychaete Perinereis aibuhitensis: Morphological changes and responses of the antioxidant system

Perinereis aibuhitensis was used to assess adverse biological effects caused by acute and chronic Pb²⁺ exposure in artificial seawater under controlled laboratory conditions. In 96-hr acute toxicity experiments, the morphological changes showed a positive time/dose-dependent tendency, and the 96-hr LC₅₀ value of Pb²⁺ was 686.41 mg/L. The responses of enzymatic and non-enzymatic antioxidants in tissues including catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA) and the content of total soluble protein (TSP), were investigated on days 1, 4, 7 and 10 after Pb²⁺ exposure under chronic toxicity testing. Results showed that the activation of the antioxidant system in P. aibuhitensis depended on the Pb²⁺ concentration and the duration of exposure time. Specifically, POD and SOD activities were induced on the first day of the exposure and decreased to the control level on day 10 after exposure. Therefore, these two indexes could be used to indicate oxidative stress associated with P. aibuhitensis exposure to Pb²⁺.