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.

     

Tolerance of ciliated protozoan Paramecium bursaria （Protozoa,Ciliophora） to ammonia and nitrites

The tolerance to ammonia and nitrites in freshwater ciliate Paramecium bursaria was measured in a conventional open system. The ciliate was exposed to different concentrations of ammonia and nitrites for 2h and 12h in order to determine the lethal concentrations. Linear regression analysis revealed that the 2h-LC50 value for ammonia was 95.94 mg/L and for nitrite 27.35 mg/L using probit scale method (with 95% confidence intervals). There was a linear correlation between the mortality probit scale and logarithmic concentration of ammonia which fit by a regression equation y=7.32x-9.51 (R^2=0.98; y, mortality probit scale; x, logarithmic concentration of ammonia), by which 2 h-LC50 value for ammonia was found to be 95.50 mg/L. A linear correlation between mortality probit scales and logarithmic concentration of nitrite is also followed the regression equation y=2.86x 0.89 (R^2=0.95; y, mortality probit scale; x, logarithmic concentration of nitrite). The regression analysis of toxicity curves showed that the linear correlation between exposed time of ammonia-N LC50 valueand ammonia-N LC50 value followed the regression equation y=2 862.85e-0.08x (R^2=0.95; y, duration of exposure to LC50 value; x, LC50 value), and that between exposed time of nitrite-N LC50 value and nitrite-N LC50 value followed the regression equation y = 127.15e-0.13x (R^2=0.91; y, exposed time of LC50 value; x, LC50 value). The results demonstrate that the tolerance to ammonia in P. bursaria is considerably higher than that of the larvae or juveniles of some metozoa, e.g. cultured prawns and oysters. In addition, ciliates, as bacterial predators, are likely to play a positive role in maintaining and improving water quality in aquatic environments with high-level ammonium, such as sewage treatment systems.     

Acute toxicity and biodegradation of endosulfan by the polychaeta Perinereis aibuhitensis in an indoor culture

The polychaete Perinereis aibuhitensis, a key species in estuarine ecosystems, can improve the culture condition of sediment. Endosulfan is an organochlorine pesticide used globally to control insects and mites; however, it is a source of pollution in aquaculture as a result of runoff or accidental release. In this study, we evaluated the toxicity of endosulfan to polychaeta and its ability to improve polluted sediment. Specifically, the effects of a series of endosulfan concentrations (0, 1.25, 2.5, 5, 10, 15, and 20 mg/L) were investigated, and the results indicated that the 24-h median lethal concentration (24-h LC₅₀) was 55.57 mg/L, while the 48-h median lethal concentration (48-h LC₅₀) was 15.56 mg/L, and the safe concentration was about 1.556 mg/L. In a 30-d exposure experiment, the animal specimen could decompose endosulfan effectively while improving endosulfan-polluted aquatic sediment.     

Toxic effects of zinc on four species of freshwater fish

The toxic effects of Zn²⁺ on Silver Carp (Hypophthalmichthyps molitrix C. et V.), Big Head Carp (Aristrichthys nobilis Richardson), Grass Carp (Ctenopharyngodon ilellus C. et V.), and Blunt Snout Bream (Megalobrama aimbly-cephala Yih) are studied. The test results are: (1) There are linear correlations between 24h LC₅₀ and 48h LC₅₀ of Zn²⁺ for Silver Carp fingerling and temperature. 24h LC₅₀, 48h LC₅₀ and 96h LC₅₀ of Zn²⁺ for fry of the four species are also determined; (2) There are logarithmic correlations between the growth rates of the fry and the concentrations of Zn²⁺ and between expansion of fish egg membranes after absorbing water and concentrations of Zn²⁺; (3) The tolerance of fry of the four species to Zn²⁺ is in the following order: Grass Carp>Silver Carp>Blunt Snout Bream>Big Head Carp; (4) The safe concentrations of Zn²⁺ are: Big Head Carp: 0.008 mg/L, Grass Carp: 0.046 mg/L, Blunt Snout Bream: 0.010 mg/L, Silver Carp: 0.012 mg/L, Silver Carp fingerling: 0.09 mg/L.     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council     

Negative effects of Phaeocystis globosa on microalgae

The potential allelopathic effects of the microalga, Phaeocystis globosa Scherffel, on three harmful bloom algae, Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Chattonella ovata Hara et Chihara were studied. The growth of C. marina and C. ovata was markedly reduced when the organisms were co-cultured with P. globosa or cultured in cell-free spent medium. Haemolytic extracts from P. globosa cells in the senescence phase had a similar inhibitory effect on the three harmful bloom algae. However, P. globosa had less influence on the brine shrimp, Artemia salina. These results indicate that P. globosa may have an allelopathic effect on microalgae, which would explain the superior competitive abilities of P. globosa. Because the addition of the haemolytic toxins from P. globosa had similar effects on algae as spent media, these compounds may be involved in the allelopathic action of P. globosa.     

Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC₅₀ at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae. Supported by the Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-YW-208), National Natural Science Foundation of China (No. 40821004, U0733006) and National Basic Research Program of China (973 Program, No. 2001CB409700)     

Variations in retention efficiency of bivalves to different concentrations and organic content of suspended particles

Retention efficiencies (RE) of scallop (Chlamys farreri), oyster (Crassostrea gigas) and mussel (Mytilus edulis) in a flow-through system were measured to understand the short-term response to various particle and organic matter concentrations. By comparing the RE of C. farreri with that of C. gigas and M. edulis, we gained further knowledge on the feeding physiological characteristics of C. farreri and ascertained the possible cause of high summer mortalities of this species. The experimental feeding conditions included natural differences in the abundance and composition of suspended seston, as well as conditions in which seston abundance and composition were manipulated by adding natural silt or cultured microalgae. The results show that in natural sea water, the minimum particle size for maximal retention in M. edulis, C. gigas and C. farreri was approximately 4, 6, and 8 μm, respectively; the RE of 2-μm (equivalent spherical diameter) particles was 17%, 19%, and 8%, respectively; and the relative standardized RE was 58%, 49%, and 18%, respectively. In C. gigas and M. edulis, the minimal particle size for maximal retention did not change with food quality (organic content). C. farreri was more sensitive to fluctuations in particle concentration and organic content. With particle concentration increase, the minimal particle size for maximal retention in scallop shifted to large particles and the RE for 2-μm particle decreased from 8% in natural seawater to 1.6%–6% in silt-enriched groups. With organic content increase, the minimal particle size for maximal retention shifted from 8 to 5 μm in natural seawater.Variation in RE of C. farreri with food conditions and the relationship between lower RE and smaller particle size may hamper C. farreri from food taking due to the decrease in the size distribution of phytoplankton in Sungo Bay.     

Lytic Characteristics and Identification of Two Alga-lysing Bacterial Strains

Lytic Characteristics and Identification of Two Alga-lysing Bacterial StrainsBeaulieu, S. E., M. R. Sengco, and D. M. Anderson, 2005. Using clay to control harmful algal blooms: deposition and resuspension of clay/algal flocs. Harmful Algae . (4): 123-138…     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Study on the phytoplankton · in a large reservoir

A comprehensive study on the community structure and function of the phytoplankton in Taipinghu Reservoir, the largest reservoir (9400 ha) in Anhui Province, China, was carried out during 1985–1986. A total of 175 species of algae belonging to 8 phyla and 87 genera was noted. The composition of phytoplankton was dominated by species of Chlorophyta, Cyanophyta and Bacillariophyta. The species number, cell density and biomass of the three groups were respectively 88%, 86.5% and 78.9% of the total phytoplankton. The weighted annual average biomass was 1.52 mg/L and cell density was 1.43×10⁶ ind/L. The growth maximum was observed in summer. Diatoms were abundant in the region adjoining a river. A large number of flagellated algae such asEuglena andChromulina occurred in the artificial fish culture bay. It can be inferred from the algal composition and total nitrogen concentration (1.48 mg/L) that this reservoir is a mesotrophic water body that had undergone slight natural eutrophication. Presented are an equation for the relation between cell density and biomass of algae and a newly developed method based on the energy flow principle in ecology for theoretically estimating algal fishery potential. The algae volumes of 128 species were measured for calculating the biomass.     

Biochemical response of the mussel Mytilus coruscus (Mytiloida: Mytilidae) exposed to in vivo sub-lethal copper concentrations

Many aquatic organisms are negatively affected by exposure to high copper concentrations.We investigated the biochemical response of the mussel Mytilus coruscus(Mytiloida:Mytilidae) to copper exposure.In vivo bioassays using M.coruscus and different copper concentrations were conducted.The activity of six biomarkers,namely superoxide dismutase(SOD),catalase(CAT),acid phosphatase(ACP),alkaline phosphatase(AKP),glutamic-oxaloacetic transaminase(GOT) and glutamic-pyruvic transaminase(GPT) were measured.Survival rates decreased with increased copper concentrations and exposure times.The LC50 values at 48,72,and 96 h exposure were 0.48,0.37,and 0.32 mg/L,respectively.Within digestive glands,CAT activity increased with increasing Cu concentrations.The activity of AKP showed no significant change,while the remaining four enzymes showed decreasing activity with increasing Cu concentrations.Within the gills,AKP activity increased when the Cu concentration was 0.05 mg/L,but showed no significant changes at higher concentrations.Activity of CAT and ACP within gills tended to decrease with increasing Cu concentration.The activity of SOD and GPT decreased at an exposure concentration of 0.2 mg/L.GOT activity within gills decreased at 0.1 mg/L and increased at an exposure concentration of 0.2 mg/L.Within the adductor muscle,AKP activity increased at 0.05 mg/L but did not change at higher exposure concentrations.ACP activity within adductor muscle tissue showed no change,while activities of CAT,GOT and GPT decreased with increasing Cu concentrations.SOD activity within the adductor muscle tissue significantly decreased at the 0.02,0.05 and 0.2 mg/L exposure concentrations.Our results show tissue specific differences for the six biomarkers in for M.coruscus.Our findings provide the basis for the establishment of reference activity levels against which biomarker changes can be estimated,and are essential preliminary steps in development of in vivo bioassays.     

Mercury (Hg2+) effect on enzyme activities and hepatopancreas histostructures of juvenile Chinese mitten crab Eriocheir sinensis

We studied the effects of mercury (Hg²⁺) on antioxidant and digestive enzyme activities in terms of LC₅₀ value and on hepatopancreas histostructures of juvenile Chinese mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg²⁺ (0, 0.01, 0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) significantly increased in the concentrations of 0.01 and 0.05 mg/L, while that of enzyme decreased in 0.10, 0.20 and 0.30 mg/L treatments. Meanwhile, Hg²⁺ disrupted the histostructures of the hepatopancreas, causing decreases in activities of pepsin, tryptase, amylase, and cellulose, which are synthesized in the hepatopancreas. Moreover, as the Hg²⁺ concentration increased, the survival rate of the crabs decreased, worst at 56.57% in 0.30 mg/L. Therefore, although crabs are able to tolerate low levels of mercury pollution, high levels lead to cellular injury and tissue damage in hepatopancreas, which then loses some of its vital physiological functions such as absorption, storage, and secretion.     

Effect of nitrogen to phosphorus ratios on cell proliferation in marine micro algae

The ratio of nitrogen/phosphorus (N/P) is known to affect cell proliferation of some marine micro algae. We evaluated the effect of N/P ratios on the proliferation and succession of phytoplankton using five marine micro algae species. We used two sources of nitrogen, NH₄Cl (N₁) and urea (N₂), and a single source of phosphorous, NaH₂PO₄(P). The optimal N/P ratio that differed among the five species was affected by the source of nitrogen, being as follows (N₁/P, N₂/P in order): Thalassiosira sp. (30/1, 20/1), Heterosigma akashiwo (30/1, 30/1), Chroomonas salina (20/1, 30/1), Chaetoceros gracilis (40/1, 60/1), and Alexandrium sp. (10/1, 30/1). Thus, the source of nitrogen must be considered when analyzing the N/P ratio. Our results provide insight for predicting phytoplankton succession in coastal waters and may be used to forecast the potential risk of harmful algal blooms.     

Toxicity of five phenolic compounds to brine shrimp Artemia sinica (Crustacea: Artemiidae)

The acute toxicity of five phenolic compounds each to 15 d old Artemia sinica was determined in this study. The brine shrimp A. sinica was hatched from the encysted dry eggs(Bohai Bay Brand) produced by Dongying Ocean Artemia Co., Ltd., China at 27℃± 1℃ in pre-filtered(through pores of 0.45 μm in diameter) and autoclaved seawater(salinity 31, pH 7.5–8.0) in a cilindroconical glass beaker(2000 mL in volume) under continuous illumination(provided by a side set 20 W fluorescent lamp) with slight aeration. Ten Artemia individuals from the same batch of the hatched were cultured in 10 mL toxicant solution prepared with seawater(salinity 31, pH 7.5–8.0) at room temperature(about 20℃) to determine 24 h, 48 h and 72 h medium lethal concentration(LC50) of 5 phenolic compounds each. It was found that the toxicity of n-heptylphenol was the highest followed by nonylphenol, t-butylphenol, 2,4-dichlorophenol and bisphenol A in order. The LC50 values of the 5 compounds were calculated with regression analysis. The real concentration(in μg- L1) of 5 phenolic compounds each in toxicant solutions was measured with GC/MS analysis. Significant loss of phenolic compounds caused by either adsorption or desorption was not found. The significant difference of LC50 values was found among the five compounds 3 exposure times each. The range between the highest no-observed-effect concentration(NOEC) and 100% death causing concentration of five phenolic compounds each was determined. The toxicity in term of 24 h LC50 value of n-HP was 9.10 times higher than that of BPA, 1.71 times higher than t-BP, 1.53 times higher than 2,4-DCP and 1.36 times higher than NP, respectively.     

Algicidal activities of secondary metabolites of marine macroalgal-derived endophytic fungi

Red tide occurrs frequently and causes signi?cant damage to the environment and human health. As a result, development of new effcient and environment friendly red-tide microalgae inhibitors has gained increasing attention in recent times. Algicolous endophytic fungi with unique habitats are promising sources for active agents owing to their abundant secondary metabolites and distinguished activities. In this study, the algicidal activities of 49 marine macroalgal-derived endophytic fungi against phytoplankton Alexandrium tamarense, Prorocentrum donghaiense, Heterosigma akashiwo, and Chattonella marina were examined using 96-well microplate. Four fungal strains, including Aspergillus wentii(pt-1), A. ustus(cf-42),and A. versicolor(dl-29, pt-20), exhibited potent algicidal activities. A total of 32 pure compounds isolated from these fungi were noted to possess dif ferent degrees of algicidal activities. Of those, 11 compounds comprising ?ve anthraquinones, two terpenoids, and four steroids showed high 24-h inhibition rates for the four red tide algae, with 24 h EC_(50) values ranging from 0.01 to 14.29 μg/mL. Among them, compound1(1,5-dihydroxy-3-methoxy-7-methylanthraquinone) presented the strongest activity against H. akashiwo,and could decrease its chlorophyll a(Chl a) and superoxide dismutase contents and increase the soluble protein, malondialdehyde, and peroxidase contents. These results suggested that the identi?ed anti-algal compound might inhibit the growth of red tide algae by weakening photosynthesis(reducing Chl a content),destroying cell membrane, and damaging the antioxidant system.     

Effects of iron electrovalence and species on growth and astaxanthin production of Haematococcus pluvialis

To increase the cell concentration and the accumulation of astaxanthin in the cultivation of Haematococcus pluvialis, effects of different iron electrovalencies (Fe²⁺-EDTA and Fe³⁺-EDTA) and species (Fe-EDTA, Fe(OH)_x^32x and FeC₆H₅O₇) addition on cell growth and accumulation of astaxanthin were studied. Results show that different iron electrovalencies have various effects on cell growth and astaxanthin accumulation of H. pluvialis. Compared with Fe³⁺-EDTA, Fe²⁺-EDTA stimulate more effectively the formation of astaxanthin. The maximum astaxanthin content (30.70 mg/g biomass cell) was obtained under conditions of 18 μmol/L Fe²⁺-EDTA, despite the lower cell density (2.3×10⁵ cell/ml) in such condition. Fe³⁺-EDTA is more effective than Fe²⁺-EDTA in improving the cell growth. Especially, the maximal steady-state cell density, 2.9×10⁵ cell/ml was obtained at 18 μmol/L Fe³⁺-EDTA addition. On the other hand, all the various species of iron (EDTA-Fe, Fe(OH)_x^32x, FeC₆H₅O₇) are capable to improve the growth of the algae and astaxanthin production. Among the three iron species, FeC₆H₅O₇ performed the best. Under the condition of a higher concentration (36 μmol/L) of FeC₆H₅O₇, the cell density and astaxanthin production is 2 and 7 times higher than those of iron-limited group, respectively. The present study demonstrates that the effects of the stimulation with different iron species increased in the order of FeC₆H₅O₇, Fe(OH)_x^32x and EDTA-Fe. Supported by Xiamen Scientific and Technologic Projects (No. 3052Z20031086), Xiamen University Alumni Association Foundation in Singapore, and the First National College Students Innovative Experimental Project     

Studies on culture conditions of benthic diatoms for feeding abalone

Effects of salinity, pH, nitrogenous and phosphate nutrients on the growth rate of four species of benthic diatoms were studied by using the method of in situ optical density measurement. The optimal culture conditions for the four species of diatoms are as follows: forAmphora coffeaeformis, s 35; pH 8.5; NO₃ ⁻−N 1.54, NH₄ ⁺−N, 7.0; CO (NH₂)₂−N, 2.5 (mg/L); PO₄ ³⁻−P 1.13 (mg/L); forCocconeis scutellum varparva, s 30; pH 8.5; NO ₃ ⁻ −N 3.08, NH ₄ ⁺ −N 3.5, CO(NH₂)₂−N 5.0 (mg/L); pO ₄ ³⁻ −P 0.283 (mg/L); forNavicula corymbosa s 25; pH 8.0; NO ₃ ⁻ −N 1.54, NH ₄ ⁺ −N 3.5 CO(NH₂)₂−N 5.0 (mg/L); PO ₄ ³⁻ −P 0.565 (mg/L); forNavicula mollis. s 25; pH 8.0; NO ₃ ⁻ −N 1.54, NH ₄ ⁺ −N 1.75, CO(NH₂)₂−N 1.25 (mg/L); PO ₄ ³⁻ −P 0.141 (mg/L). Part of the results of this paper were exchanged in the Fourth Asian Aquaculture Forum. Oct. 16–20. 1995. Beijing, P. R. China.     

Exploration of Antifouling Potential of the Brown Algae <Emphasis Type="Italic">Laminaria</Emphasis> ‘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.     

Effects of nutrients and light intensity on the growth and biochemical composition of a marine microalga Odontella aurita

Algal biotechnology has advanced greatly in the past three decades. Many microalgae are now cultivated to produce bioactive substances. Odontella aurita is a marine diatom industrially cultured in outdoor open ponds and used for human nutrition. For the first time, we have systematically investigated the effects of culture conditions in cylindrical glass columns and fiat-plate photobioreactors, including nutrients （nitrogen, phosphorus, silicon, and sulfur）, light intensity and light path, on O. aurita cell growth and biochemical composition （protein, carbohydrate, β-1,3-glucan, lipids, and ash）. The optimal medium for photoautotrophic cultivation of O. aurita contained 17.65 mmol/L nitrogen, 1.09 mmol/L phosphorus, 0.42 mmol/L silicon, and 24.51 mmol/L sulfur, yielding a maximum biomass production of 6.1-6.8 g/L and 6.7-7.8 g/L under low and high light, respectively. Scale-up experiments were conducted with fiat-plate photobioreactors using different light-paths, indicating that a short light path was more suitable for biomass production of O. aurita. Analyses of biochemical composition showed that protein content decreased while carbohydrate （mainly composed of 15-1,3-glucan） increased remarkably to about 50% of dry weight during the entire culture period. The highest lipid content （19.7% of dry weight） was obtained under 0.11 mmol/L silicon and high light conditions at harvest time. Fatty acid profiles revealed that 80% were Cx4, C~6, and C20, while arachidonic acid and eicosapentaenoic acid （EPA） accounted for 1.6%-5.6% and 9%-20% of total fatty acids, respectively. High biomass production and characteristic biochemical composition profiles make O. aurita a promising microalga for the production ofbioactive components, such as EPA and D-1,3-glucan.