The effects of nitrogen,phosphorus, vitamins and trace metals on the growth of the red tide organismProrocentrum Micans

Laboratory culture experiments showed that <100μ mol/L nitrate, amonium or mixture of amino acids promote the growth of the red tide organismProrocentrum micans Ehrenb, but that >100μmol/L of ammonium, or mixture of glycine and glutamate was harmful to growth, and that orthophosphate wasP. micans’ main phosphorous source in the ocean. Presence of 80μ mol/L EDTA, 0.5 to 1 μmol/L Fe³⁺, 1.0 to 20.0 μ mol/L Mn²⁺ 0.1 to 0.4 μmol/L Co²⁺ in the culture medium could improve the growth ofP. micans. Vitamin B₁ promoted growth, but vitamin B₁₂ and biotin did not. The estimated minimum cell quotas (q _o) for nitrogen and phosphorus being 0.74 pmole/cell and 0.045 pmole/cell show that phosphorus (more than nitrogen) limits the growth ofP. micans in the study area. This project was supported by the Natural Science Foundation of Zhejiang Province.     

Bioaccumulation of Pb<Superscript>2+</Superscript> and its effects on growth,morphology and pigment contents of <Emphasis Type="Italic">Spirulina</Emphasis> (<Emphasis Type="Italic">Arthrospira</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis>

A laboratory experiment was conducted to assess the bioaccumulation of Pb²⁺ and its effects on growth, morphology and pigment contents of Spirulina (Arthrospira) platensis. The specimen cultured in Zarrouk liquid medium was treated with various initial metal concentrations (0, 5, 10, 30, 50 and 100 μg mL⁻¹). The growth of S. platensis was adversely affected by Pb²⁺ at high concentrations (30, 50 and 100 μg mL⁻¹). However, at low concentrations (5 μg mL⁻¹), Pb²⁺ could stimulate its growth slightly. The pigment contents (chlorophyll α and β carotene) were decreased in a dose-dependent manner. The highest reductions (67% and 53% respectively in chlorophyll α and β carotene) were observed in 100 μg mL⁻¹ treatment group. The LC₅₀ (96 h) of Pb²⁺ was measured as 75.34 μg mL⁻¹. Apart from a few cases of filament breakages at elevated concentrations (50 and 100 μg mL⁻¹), morphological abnormalities are not specific. Metal bioaccumulation increased with Pb²⁺ concentrations, but decreased with exposure time. The maximum accumulated amount was 188 mg g⁻¹ dry weight. The bioconcentration factor (BCF) reached to a peak at day 2, followed by a gradual reduction for all the exposure concentrations. S. platensis is able to tolerate considerably high Pb²⁺ concentrations. Consequently it can be used as a potential species to remove heavy metal from contaminated waters.     

Effects of nutritional factors on the growth and heterotrophic eicosapentaenoic acid production of diatom <Emphasis Type="Italic">Nitzschia laevis</Emphasis>

The effects of several nutritional factors on the growth and eicosapentaenoic acid (EPA) production of diatom Nitzschia laevis were studied. 4 LDM (quadrupled concentration of the nutrient salt) was the optimal concentration of nutrient salt for the growth and EPA production of N. laevis. The growth of N. laevis was inhibited when the glucose concentration was either lower than 10 gL⁻¹ or higher than 15 gL⁻¹. Both sodium nitrate and urea were good nitrogen sources for the growth and EPA production, while ammonium chloride seriously decreased the dry cell weight (DW) and the EPA content. Silicate seriously influenced the growth of N. laevis. The maximum DW of 2.34 gL⁻¹ was obtained in the presence of 150 mgL⁻¹ Na₂SiO₃·9H₂O. The EPA content remained almost the same when the silicate concentration was lower than 150 mgL⁻¹; however, higher silicate concentrations resulted in a steady decrease of EPA content. Low medium salinity (⩽29) did not seem to influence the DW of N. laevis, and high salinity resulted in a decrease of DW. The highest EPA content (4.08%) and yield (110 mgL⁻¹) were observed at the salinity of 36 and 29, respectively.     

On the ultrastructure and classification of the hemocytes of penaeid shrimp,Penaeus vannamei (Crustacea, Decapoda)

The ultrastructure of hemocytes ofPenaeus vannamei was examined by transmission electron microscope. Three types of hemocytes were identified: hyaline cell, small-granule cell and large-granule cell. Hyaline cells are the smallest of the hemocytes, lack electron-dense granules, have average size of 11.11 μm (SE±0.50,n=10)×6.96μm (SE±0.55,n=10), comprise 25.8% (SE±2.87,n=4×10) of the hemocytes; small granule cells are the most abundant type of hemocytes, comprise 58.1% (SE±3.40,n=4×10) of the hemocytes, contain small electron—dense granules, have average size of 10.78 μm (SE±0.60,n=10)×8.63 μm (SE±0.44,n=10); and large-granule cells comprise 16.1% (SE±2.55,n=4×10) of the hemocytes, are filled with large electron dense granules, have average size of 12.51 μm (SE±0.63,n=10)×8.99μm (SE±0.71,n=10). This research was supported by the National Climbing Project B, PDB6-6-3. Contribution No. 3448 from the Institute of Oceanology, Chinese Academy of Sciences.     

Petroleum Hydrocarbon Distribution Features in Water and Sediment off Fujian Shore

This study on the distribution features of petroleum hydrocarbon in water and sedi-ment off the Fujian shore using data obtained from the baseline research on oceanic contamination in Fujian showed that:during the research preiod,petroleum hydrocarbon concentrations in water varied from 5.77μg/L to 37.28μg/L,averaged 14.48 μg/L;was lower in the wet seasson than in the dry season;and was highest in the Minjiang Estuary and Jiulong Estuary in both seasons ,The petorleum hydrocarbon concentrations in shore sediment varied from 14.48 mg/kg to 784.36 mg/kg,averaged 133.3 mg/kg ,and was closely related to sediment types (granularity).     

Preparation and characteristics of biosilica derived from marine diatom biomass of Nitzschia closterium and Thalassiosira

In this study, biosilica of high purity was successfully prepared from marine diatom( Nitzschia closterium and Thalassiosira) biomass using an optimized novel method with acid washing treatment followed by thermal treatment of the biomass. The optimal condition of the method was 2% diluted HCl washing and baking at 600°C. The SiO_2 contents of N. closterium biosilica and Thalassiosira biosilica were 92.23% and 91.52%, respectively, which were both higher than that of diatomite biosilica. The SiO_2 morphologies of both biosilica are typical amorphous silica. Besides, N. closterium biosilica possessed micropores and fibers with a surface area of 59.81 m~2/g. And Thalassiosira biosilica possessed a mesoporous hierarchical skeleton with a surface area of 9.91 m~2/g. The results suggest that the biosilica samples obtained in this study present highly porous structures. The prepared porous biosilica material possesses great potential to be used as drug delivery carrier, biosensor, biocatalyst as well as adsorbent in the future.     

The influence of Fe^2＋ on growth and development of cells enzymatically isolated from Porphyra yezoensis blades

Fe^2＋ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe^2＋ requirement in nori had not been seen. Porphyra yezoensis cells were separated enzymatically and cultured in a series of sterilized seawater media containing various concentrations of Fe^2＋. The growth development and cell were investigated in this work. Through this experiment, two biologically-meant concentration scales were found, one is low concentrations, 12.1-102.1μg/L, 10-100 times than that in seawater, favoring the development of isolated cells of Porphyra and the other was high concentrations, more than 10mg/L inhibiting the cell growth, leading to the deformity and shrinkage of the cells. At the concentration of 50 mg/L, the cells stopped growing and died eventually.     

Impacts of CO<Subscript>2</Subscript> enrichment on growth and photosynthesis in freshwater and marine diatoms

The physiological responses of Nitzschia palea Kvtzing, a freshwater diatom, to elevated CO2 were investigated and compared with those of a marine diatom, Chaetoceros muelleri Lemmermann previously reported. Elevated CO2 concentration to 700 μl/L increased the dissolved inorganic carbon （D!C） and lowered the pH in the cultures of N. palea, thus enhancing the growth by 4%-20% during the whole growth period. High CO2-grown N. palea cells showed lower levels of dark respiration rates and higher Ik values. Light-saturated photosynthetic rates and photosynthetic efficiencies decreased in N. palea with the doubling CO2 concentration in airflow to the bottom of cultures, although the doubling CO2 concentration in airflow to the surface cultures had few effects on these two photosynthetic parameters. N. palea cells were found to be capable of using HCO5 in addition to gaseous CO2, and the CO2 enrichment decreased their affinity for HCO5 and CO2. Although doubled CO2 level would enhance the biomass of N. patea and C. muelleri to different extents, compared with the marine diatom, it had a significant effect on the specific growth rates of N. palea. In addition, the responses of photosynthetic parameters of IV. palea to doubled CO2 concentration were almost opposite to those of C. muelleri.     

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.     

Effects of Mg2+, NaCl, citric acid, and other factors on synthesis and accumulation of β-carotene inDunaliella salina

The effect of Mg²⁺, NaCl and citric acid on the accumulation of β-carotene inDunaliella Salina was studied. The experimental results showed that 10.5 mmol/L Mg²⁺, 5 mol/L NaCl, 3 μmol/L citric acid, and CO₂ are favorable forDunaliella Salina cell growth and β-carotene accumulation. After 144 h culture under the above conditions, theDunaliella Salina biomass increased by 7.18 times; β-carotene reached 9.61%. Project 89023990 supported by the Shandong Natural Science Fund.     

The inhibitory degree between Skeletonema costatum and dinoflagllate Prorocentrum donghaiense at different concentrations of phosphate and nitrate/phosphate ratios

Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate (N/P) ratios. Experiments were conducted under P-limited conditions and the Lotka-Volterra mathematical model was used to simulate the growth of S. costatum and P. donghaiense in the bi-algal cultures. Both of these two species were inhibited significantly in bi-algal culture. The results of the simulation showed that the inhibitory degree of S. costaum by P. donghaiense was high when the concentration of PO4-P was low (0.1μmolL-1/2 d), but that of P. donghaiense by S. costaum was high with increased PO4-P supply (0.6μmolL-1/2 d). At low concen-tration of PO4-P (0.1μmolL-1/2 d), or high concentration of PO4-P (0.6μmolL-1/2 d) with high N/P ratio (160), the interactions be-tween S. costatum and P. donghaiense were dependent on the initial cell densities of both species. At high concentration of PO4-P (0.6μmolL-1/2 d) with low N/P ratio (25 or 80), S. costatum exhibited a survival strategy superior to that of P. donghaiense. The de-gree of inhibition of P. donghaiense by S. costaum increased with elevated N/P ratio when the medium was supplemented with con-centration 0.1μmolL-1/2 d of PO4-P. The degree of inhibition to P. donghaiense by S. costaum increased with elevated N/P ratio at low concentration of PO4-P (0.1 μmolL-1/2 d). This trend was conversed at high concentration of PO4-P (0.6μmolL-1/2 d). However, the degree of inhibition of S. costaum by P. donghaiense increased with the increased N/P ratio at different PO4-P concentrations (0.1μmolL-1/2 d and 0.6μmolL-1/2 d). These results suggested that both phosphate concentration and N/P ratio affected the competition between S. costaum and P. donghaiense: P. donghaiense is more competitive in environments with low phosphate or high N/P ratio and the influence of N/P ratio on the competition was more significant with lower phosphate concentration.     

ORGANIC GEOCHEMISTRY MARKERSOF CLIMATIC EVOLUTION IN LOESSREGION OF CHINA

Two hundred and thirty-five samples collected from Weinan loess section (L₂ – S₀) have been analyzed by TOC (total organic carbon) and pyrolysis hydrocarbons. The curve of TOC with depth has been set up and compared well with other climatic markers. The main periods of TOC variation of the last 0.14 Ma are 67.08 ka and 18.29 ka, which is near with the periods of the variation of sunshine, showing that the TOC variation may be controlled by the variation of 65°N sun-shine. The distribution of pyrolysis hydrocarbon and its parameters shows that there are two kinds of hydrocarbon associated with climatic change. One is called hydrocarbon pyrolysted directly from organism (pHC_2–1) which is in the range of T _max350°C – 500°C, and associated with atmospheric water. The other is called carbonate inclusion hydrocarbon (pHC_2–2) which is in the range of T _max 500°C – 600°C and formed by evaporation and associated with dry climate. The formula calculating humidity has been deduced Og=pHC_2–1/(pHC_2–1 + pHC_2–2). The range of humidity of the section is 47% – 77% and the reducing order is S_1–3, S_1–2, S₀, S_1–1, S_1–4, S_1–1, L_1–1, L_1–3, and L_1–5. And the trend of the humidity is still increasing today.     

In-situ studies of microzooplankton grazing pressure on phytoplankton in Jiaozhou Bay, China

Preliminary studies on microzooplankton grazing were conducted with dilution method in Jiaozhou Bay from summer 1998 to spring 1999. Four experiments were carried out at St. 5 located at the center of Jiaozhou Bay. Chlorophyll a concentrations were consistently dominated by netphytoplankton (net-, >20μm), except during the autumn 1998 cruise, when they were dominated by nanophytoplankton(nano-, 2–20μm). The contribution of picophytoplankton (pico-, <2μm) to total chlorophyll a concentrations (<200μm) varied considerably between cruises. Instantaneous growth coefficients(u) of phytoplankton varied from 0.098 to 1.947d⁻¹, with mean value of 0.902d⁻¹. Instantaneous coefficients(g) of microzooplankton grazing on phytoplankton ranged from 0.066 to 0.567d⁻¹, mean value of 0.265d⁻¹, which was equivalent to daily lose of 21.9% of the initial standing stock and 58.1% of the daily potential production. Project No KZCX3-SW-214 supported by Chinese Academy of Sciences.     

Effects of Body Weight and Water Temperature on Maximum Food Consumption of Juvenile Sebastodes fuscescens （Houttuyn）

Maximum rate of food consumption ( Cmax ) was determined for juvenile Sebastodes fuscescens (Houttuyn) at water temperature of 10, 15, 20 and 25℃. The relationships of Cmax to the body weight (W) at each temperature were described by a power equation: lnCmax = a b lnW. Covariance analysis revealed significant interaction of the temperature and body weight. The relationship of adjusted Cmax to water temperature (T) was described by a quadratic equation: Cmax = -0.369 0.456T - 0.0117T^2. The optimal feeding temperature calculated from this equation was 19.5 ℃. The coefficients of the multiple regression estimation relating Cmax to body weight (W) and water temperature (T) were given in the Table 2.     

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     

Effect of cytokinins and auxins on the growth of free-living conchocelis ofPorphyra yezoensis

IAA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis ofPorphyra yezoensis which showed that its growth in medium with cytokinins, IAA and NAA was more rapid than that in medium with non—phytohormones; that the optimal concentrations for promoting growth were 10 μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, IAA and cytokinins, NAA/ZA 1–1000/1 μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. IAA/ZA 0.1-1/0.1-1 μg/L; IAA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000 μg/L also promoted growth. Contribution No. 2335 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 94 from the Experimental Marine Biology Lab Qingdao, China. The research was supported by a grant from the Chinese Academy of Sciences.     

DYNAMIC CHANGES OF INORGANIC NITROGEN AND ASTAXANTI—IIN ACCUMULATION IN HAEMATOCOCCUS PLUVIALIS

This study on dynamic changes of culture color,astaxanthin and chlorophylls,inorganic N including N-NO3^-,N-NO2^- and N-NH4^ in batch culture of Haematococcus pluvialis exposed to different additive nitrate concentration showed(1)ast/chl ratio was over 0.8 for brown and red algae,but was usually less than 0.5 for green and yellow algae;(2)N-NO3^-,in general,was unstable and decreased,except for a small unexpected increase in nitrate enriched treatment groups;(3)measurable amounts of N-NO2^- and N-NH4^ were observed respectively with three change modes although no extemal nitrite and ammonia were added into the culture;(4)a non-linear correlation between ast/chl ratio(or color)changes and the levels of N-NO3^-,N-NO2^-,N-NH4^ in H.pluvialis culture;(5)up and down variation of the ast/chl ratio occurred simultaneously with a perceptible color change from yellow to brown(or red)when N-NO3^-,N-NO2^- and N-NH4^ fluctuated around 30,5,5μmol/L respectively;(6)existence of three dynamic modes of N-NO3^-,N-NO2^- and N-NH4^ changes,obviously associated with initial extemal nitrate;(7)the key level of total inorganic N concentration regulating the above physiological changes during indoor cultivation was about 50 μmol/L;and(8)0.5-10mmol/L of nitrate was theoretically conducive to cell growth in batch culture.     

Water quality and eutrophication in the Guangzhou Sea Zone of the Pearl River estuary

To gain a better understanding of water quality and eutrophication, we investigated the seasonal and spatial distribution of water quality at 17 stations in the Guangzhou Sea Zone (GZSZ). Nutrients, chlorophyll-a (Chl-a), salinity, chemical oxygen demand, and other physical and chemical parameters were determined in February, May, August and October from 2005 to 2007. The concentrations showed ranges of 93.2–530.4 μmol/L for dissolved inorganic nitrogen (DIN), 0.62–3.16 μmol/L for phosphate (PO₄-P) and 50–127 μmol/L for silicate (SiO₃-Si). The results indicated that DIN was strongly influenced by domestic sewage coming from Guangzhou City and NO₃-N was the main form of DIN in most areas, while concentrations of phosphate and silicate were generally controlled by Pearl River runoff, land-based sources along the land or islands, and algae assimilation. N/P ratios were very high in both dry season and wet season, and varied from 57 to 667, suggesting that P was potentially the most limiting nutrient in the GZSZ. The concentrations of Chl-a were 3–96 μg/L, and were highly correlated with the distribution characteristics of COD. The concentrations of nutrients have increased over the past two decades (1982–2007). This means it is necessary to control the input of nutrients to the GZSZ, especially that of phosphate.