COPPER ACCUMULATION AND TRANSPORT IN A MARINE FOOD CHAIN COMPOSED OF PLATYMONAS SUBCORDIFORMIS, BRACHIONUS PLICATILIS AND PENAEUS MONODON

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Relationship between heavy metal pollution and water productivity in Xiamen estuarine harbor

This paper is based on the results of simulation experiments and annual observations on the effect of Cr, Cu, Hg, and Zn on diatom (Chaetoceros sp.Skeletonema costatum andMelosira sulcata) growth. Laboratory experiments reveal many physico-chemical parameters play an important role in the toxicity and accumulation of metals in organisms. The mesotrophic level in the water environment is related to phytoplankton growth. A higher nutrient level can have a negative impact on diatom production. Several of the above metals coexist in the investigated region and exert mainly a negative effect on growth. This paper was published in Chinese in theOceanologia et Limnologia Sinica 17(3):173–183, 1986.     

Accumulation of petroleum hydrocarbons and heavy metals in clams （Ruditapes philippinarum） in Jiaozhou Bay, China

Accumulation and distributions of aliphatic and polyaromatic hydrocarbons (PAHs) and heavy metals were measured in tissues of the clam Ruditapes philippinarum collected from 5 sites in Jiaozhou Bay, Qingdao, China. The concentrations of total aliphatic hydrocarbon and PAHs ranged from 570 to 2 574 ng/gdw (gram dry weight) and from 276 to 939 ng/gdw, in the most and least polluted sites, respectively. The bio-accumulation of hydrocarbons and PAHs in the clams appeared to be selective. Aliphatic hydrocarbons were predominantly represented by short chain (<nC₂₃) n-alkanes, suggesting that petroleum hydrocarbons were likely the major contamination source. The selective uptake of 3 and 4 ring PAHs, such as naphthalene, fluorene, phenanthrene, fluoranthene and pyrene, by the clams was probably related to the physiological and bio-kinetic processes that were energetically favorable for uptake of compounds with fewer rings. Accumulation of the metals Cd, Cu, Zn, Pb, Cr, Hg, and As in the clam tissues also showed high variability, ranging from 0.043 to 87 μg/gdw. Among the 7 detected metals, Zn, Cd, Cu, and As had a particularly high potential of accumulation in R. philippinarum. In general, a positive correlation was found between the tissue concentrations and sediment concentrations of hydrocarbons and of some metals. Our study suggests that moderate contamination with polyaromatic hydrocarbons, and low to moderate contamination with metals, currently exists for clam R. philippinarum in Jiaozhou Bay, in comparison with other regional studies. A long-term monitoring program is certainly needed for assessment of the potential ecological influence and toxicity of these contaminants of R. philippinarum in Jiaozhou Bay.     

Effect of bis(tributyltin) oxide on reproduction and population growth rate of calanoid copepod Schmackeria poplesia

A full life-cycle toxicity test, combined with histology, on calanoid copepod Schmackeria poplesia was used to study the effect of bis(tributyltin) oxide (TBTO). The results indicate no sex-specific differences in TBTO toxicity. Long-term mortalities of the copepods exposed to concentrations higher than 20 ng TBTO L⁻¹ were significantly elevated compared with that of control, and larval development was inhibited when they were exposed to 40 and 60 ng TBTO L⁻¹. The percentages of ovigerous females were reduced compared with the control (P<0.01) after 24 days exposure to concentrations higher than 10 ng TBTO L⁻¹. Histological examinations suggest that exposure to TBTO might block the posterior end of the diverticula and inhibits the production of egg sacs. A modified Euler-Lotka equation was used to calculate a population-level endpoint, the intrinsic rate of natural increase (r _m), from individual life-table endpoints, i.e. mortality rate, time of release of first brood, sex ratio, the fraction of ovigerous females among all females as well as the number of nauplii per ovigerous female. Apart from the highest TBTO concentration (60 ng L⁻¹), where all females aborted their egg sacs, 20 ng TBTO L⁻¹ was the only concentration that significantly decreased r _m compared with that of control (an effect associated with decreased sex ratio). The results show that the S. poplesia is affected by prolonged exposure to low concentrations of TBTO. The full life-cycle toxicity test combined with histology experiments provides more integral understanding of the toxicity of endocrine disrupters.     

A model describing the time course and variability in toxicity of hydrophobic chemicals to aquatic organisms

The growth of Chlorella marine, Nannochloris oculate, Pyramimonaos sp., Platymonas subcordiformis and Phaeodactylum tricornutum exposed to chlorobenzene, 1,2-dichlorobenzene, 1,2,3,4-tetrachlorobenzene and pentachlorobenzene was tested. The Boltzman equation was used to describe organism growth. The time course for uptake of hydrophobic organic chemicals (HOCs) by aquatic organisms was expressed by incorporating growth and, if desired, the effect of metabolism into the HOC bioconcentration process. The probability of any given concentration of HOCs in the organisms causing a specified toxic endpoint was expressed with a modified Weibull distribution function. The combined bioconcentration and probability equations were tested with data for time course of incubation of algae exposed to chlorinated benzenes (CBs). A set of parameters, including the uptake rate constant k ₁, the elimination rate constant k ₂ and thereafter the bioconcentration factor on a dry weight basis, BCF _D, the critical HOC concentration in the organism resulting in a specified toxic endpoint, C _A ^* , and the spread factor, S, could be obtained by fitting only experimental data for percent growth inhibition(%)-time-CB exposure concentration. The average coefficients of variation within CBs were 15.2% for BCF _D, 21.0% for k ₁, 18.3% for k ₂, 8.1% for C _A ^* and 9.7% for S. The variability in toxicity (such as EC₁₀, EC₅₀, EC₉₀) derived from the model equations agreed well with those experimentally observed.     

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)     

Acute toxicities of potassium permanganate, formalin, and Lugol’s iodine solution to a marine ciliate, Pleuronema coronatum (ciliophora, scuticociliatida)

Acute toxicities of potassium permanganate, formalin, and Lugol’s iodine solution to a commonly occurred marine ciliate Pleuronema coronatum (Ciliophora, Scuticociliatida) were measured. Linear regression analysis of the results highlighted the close relationships between doses of the medicines and mortalities of the organisms, thus providing a capability to predict toxicity effects from the dose. Toxic effects of the medicines on the ciliates were described in the present paper, and the median lethal concentrations (LC₅₀ values) were given. Results of measurements indicated that 2 h-LC₅₀ and 12 h-LC₅₀ values of formalin on P. coronatum were 59.00×10⁻⁶ and 43.57×10⁻⁶, while those of Lugol’s solutions were 90.13 and 67.84×10⁻⁶ respectively. The tolerance of P. coronatum to formalin is apparently lower than that to Lugol’s iodine solution and potassium permanganate is a suitable medicine to kill ciliates in short time.     

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.     

Variation in Rubisco and other photosynthetic parameters in the life cycle of Haematococcus pluvialis

Cells of Haematococcus pluvialis Flot. et Will were collected in four different growth phases. We quantified the initial and total enzyme activity of ribulose-1,5-bisphosphate carboxylase (Rubisco) in crude extracts, and the relative expression of large-subunit ribulose-1,5-bisphosphate caboxylase / oxygenase (rbcL) mRNA. We measured the ratio of photosynthetic rate to respiration rate (P/R), maximal effective quantum yield of photosystem II (Fv/Fm), electron transport rate (ETR), actual photochemical efficiency of PSII in the light (ΦPSII), and non-photochemical quenching (NPQ). Green vegetative cells were found to be in the most active state, with a relatively higher P/R ratio. These cells also displayed the lowest NPQ and the highest Fv/Fm, ETR, and ΦPSII, indicating the most effective PSII. However, both Rubisco activity and rbcL mRNA expression were the lowest measured. In orange resting cysts with relatively lower P/R and NPQ, Rubisco activity and rbcL expression reached a peak, while Fv/Fm, ETR, and ΦPSII were the lowest measured. Taking into account the methods of astaxanthin induction used in industry, we suggest that Rubisco may participate in astaxanthin accumulation in H. pluvialis. A continuous and sufficient supply of a carbon source such as CO2 may therefore aid the large scale production of astaxanthin.     

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.     

Optimum temperature for the growth performance of juvenile orange-spotted grouper （Epinephelus coioides H.）

Effects of water temperature （17, 21, 25, 30 and 35℃） and body size （14.75-281.41 g initial body weight） on food consumption, growth, feed conversion, and dry matter content in orange-spotted grouper fed to satiation were investigated. The combined effect of temperature （T, ℃） and body weight （W, g） on maximum food consumption （Cmax, g/day） was described as： InCmax=-7.411＋0.828 InW＋0.317T4）.004 7T2, and the optimum feeding temperature was 33.9℃. The combined effect of temperature and body weight on growth （G） was described as： lnG=-4.461-0.2081nW＋0.394T-0.006 3T^2. The optimum growth temperature was 31.4℃, whereas overall growth rates were high at 25, 30 and 35 ℃. Feed conversion efficiencies （FCE, %）, increasing first and then decreasing with increasing temperature, averaged from 1.8 to 2.1 in terms of dry weight of food fish. The optimum temperature for FCE tended to be lower than that for growth or feeding. Dry matter content increased with both increasing water temperature （17, 25, 30 and 35℃） and body weight, and the combined effect of temperature and body weight on dry matter content （DM, %） was described as： lnDM =3.232＋0.01 4 lnW-0.004 4T＋0.001 2TInW.     

Distribution and accumulation characteristics of heavy metals in sediments in southern sea area of Huludao City, China

The southern sea area of the Huludao City, Liaoning Province might be polluted by heavy metals because it is close to the Jinzhou Bay, one of the heaviest sea area polluted by heavy metals in China. The undisturbed modern sediment core can be used to analyze the accumulation and source of the pollutants using 137 Cs and 210 Pb ex . Thirty-five samples of surface sediment and two core sediments were collected from the southern sea area of Huludao City. The concentrations of copper (Cu), lead (Pb), chrome (Cr), zinc (Zn), arsenic (As) and mercury (Hg) in the surface sediments as well as Cu, Pb, Zn, Cr, 137 Cs and 210 Pb ex in the core sediments were determined to research the spatial distribution and accumulation characteristics, and to analyze the sources and the potential risks of heavy metals. The results show that the pollution levels of Zn and Hg are serious, and 26 stations are at moderate or heavy ecological risks. The concentrations of the heavy metals increase from east to west, as well as from open sea to offshore marine area. The concentrations of heavy metals are not high in the sediments adjacent to the Jinzhou Bay, and the influence caused by the seawater exchange with the Jinzhou Bay is little. The concentrations of the heavy metals in the core sediments show low-high-low characteristic, and it coincides with the pollution history of Huludao City. The atmospheric deposition of heavy metals from the Huludao Zinc Plant is likely to be the main source of pollution without direct discharge of wastewater. The high concentrations of heavy metals appear on the upper sediment of 20 cm. The shallow sediment with high heavy metal contents might be exposed to surface when it was disturbed by the ocean engineering and big storm surge, then cause risk to the safety of aquaculture and human healthy.     

Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

Estradiol treatment during early life stages of tiger puffer Takifugu rubripes induces feminization in genetic males. However, the ovaries in genetic males may revert to testes once estradiol treatment is halted.Therefore studies should investigate molecular mechanisms underlying ovary-to-testis recovery in genetic males after treatment. In the present study, tiger puffer were exposed to 10, and 100 μg/L 17 p-estradiol(E_2)from 15 to 100 days post-hatching(dph), then gonad phenotypes and expression profiles of six sex-related genes(cyp19a,foxl2, dmrtl, amh, sox9a, and sox9b) were characterized after the exposure. Results showed that both 10 and 100 μg/L E_2 induced ovarian development in genetic males at 100 dph. However, all ovaries induced by 10 μg/L E_2 first developed into intersexual gonads and subsequently reverted to testes after the exposure. As for treatment of 100 pg/L E_2, while the rest of the ovaries maintained morphological stability,percentages of intersexual gonads reached 38%-57%, and none were reverted to testes. Increased mRNA levels of cyp19 a,foxl2 and sox9b and decreased mRNA levels of dmrtl, amh, and sox9a were observed during the ovarian development in genetic males. While contrary gene expression profiles were detected during ovary-to-testis transformation. The mRNA levels of all the six genes were increased during the development of intersexual gonads. These results indicated that up-regulation of dmrtl, amh and sox9a is associated with initial ovary-to-intersexual transformation, and suppression of foxl2, cypl9a and sox9b is essential for complete ovary-to-testis recovery in genetic males. This research will help to trace the molecular processes underlying gonadal transformation in teleosts.     

Effect of nitrate and phosphate on accumulation of β-carotene isomers inDunaliella salina

Dunaliella salina, a halotolerant unicellular green alga, can accumulate a large amount of β-carotene under certain environmental conditions. The isomers of β-carotene extracted fromD. salina cultured in medium with different nitrate and phosphate concentrations were analysed by HPLC with Alox-T alumina column. At least six isomers were found in different proportions depending on the culture media's nitrate and/or phosphate concentrations. Nitrate and/or phosphate deficiency was conducive to the accumulation of total cis isomers but not of all trans isomer. It is suggested that 1 mmol/L KNO₃ and 0.1 mmol/L KH₂ PO₄ are favourable for accumulation of total cis β-carotene. Contribution No. 2090 from the Institute of Oceanology, Chinese Academy of Sciences. This project was supported by the National Natural Science Foundation of China, Grant No. 38970587.     

Effects of calanoid copepod Schmackeria poplesia as a live food on the growth, survival and fatty acid composition of larvae and juveniles of Japanese flounder, Paralichthys olivaceus

Zooplankton constitutes a major part of the diet for fish larvae in the marine food web, and it is generally believed that copepods can meet the nutritional requirements of fish larvae. In this study, calanoid copepod Schmackeria poplesia, rotifer Brachionus plicatilis and anostraca crustacean Artemia sp. were analyzed for fatty acid contents, and were used as live food for culturing larval Japanese flounder, Paralichthys olivaceus. The total content of three types of HUFAs (DHA, EPA and ARA) in S. poplesia was significantly higher than that in the other two live foods (P<0.01). Three live organisms were used for raising larvae and juveniles of Paralichthys olivaceus respectively for 15 and 10 d. Then the growth, survival and fatty acid composition of the larvae and juveniles were investigated. The results showed that the larvae and juveniles fed with copepods (S. poplesia) had significantly higher growth rate than those fed with the other two organisms (P<0.01). The survival of the flounder larvae fed with copepods was significantly higher than that of the others (P<0.01), and the survival of the juvenile fish fed with copepods was higher than that fed with Artemia (P<0.05). The contents of three types of HUFAs (DHA, EPA and ARA) and the ratio of DHA/EPA in larval and juvenile flounder P. olivaceus were analyzed. The results showed that the contents of DHA, EPA and ARA in the larvae and juveniles fed with S. poplesia were higher than those fed with a mixed diet or Artemia only, and the ratio of EPA/ARA in larvae and juveniles of P. olivaceus fed with S. poplesia was lower than that in the case of feeding with a mixed diet or Artemia only. The present data showed that copepod is the best choice for feeding the larvae and juveniles of fish considering its effects on the survival, growth and nutrition composition of the fish.     

Effect of zinc on the growth and development of larvae of bay scallopArgopecten Irradians

The 48-h straight-hinge larvae ofArgopecten irradians were exposed for 9 days to various concentrations of zinc in seawater. The growth rate of larvae in the 50 ppb Zn mediums was 77% that of the controls, and nearly zero in the 200 ppb Zn mediums. A progressive decrease in growth and increase in larval deformity and mortality was observed with increasing zinc concentrations from 50 to 200 ppb. 50 and 100 ppb Zn resulted in delayed development of eye spot and metamorphosis and reduced numbers of larvae at both stages. All larvae subjected to higher zinc concentrations died before attaining the eye-spot stage. The zinc concentrations which caused 50% reduction in growth (G C₅₀) and 50% mortality (LC₅₀) were 109 and 120 ppb respectively. Growing in the medium with 100 ppb Zn, the larvae fed withIsochrysis galbana cultured in a medium with 100 ppb Zn showed more suppressed growth and development than those fed with normal food. It is suggested that zinc contaminated food has strong influence on the bay scallop larvae. Contribution No. 1616 from the Institute of Oceanology, Academia Sinica.     

Fractal dimensions of flocs between clay particles and HAB organisms

The impact of harmful algal blooms (HABs) on public health and related economics have been increasing in many coastal regions of the world. Sedimentation of algal cells through flocculation with clay particles is a promising strategy for controlling HABs. Previous studies found that removal efficiency (RE) was influenced by many factors, including clay type and concentration, algal growth stage, and physiological aspects of HAB cells. To estimate the effect of morphological characteristics of the aggregates on HAB cell removal, fractal dimensions were measured and the RE of three species of HAB organism, Heterosigma akashiwo, Alexandrium tamarense, and Skeletonema costatum, by original clay and modified clay, was determined. For all HAB species, the modified clay had a higher RE than original clay. For the original clay, the two-dimensional fractal dimension (D2) was 1.92 and three-dimensional fractal dimension (D3) 2.81, while for the modified clay, D2 was 1.84 and D3 was 2.50. The addition of polyaluminum chloride (PACl) lead to a decrease of the repulsive barrier between clay particles, and resulted in lower D2 and D3. Due to the decrease of D3, and the increase of the effective sticking coefficient, the flocculation rate between modified clay particles and HAB organisms increased, and thus resulted in a high RE. The fractal dimensions of flocs differed in HAB species with different cell morphologies. For example, Alexandrium tamarense cells are ellipsoidal, and the D3 and D2 of flocs were the highest, while for Skeletonema costatum, which has filamentous cells, the D3 and D2 of flocs were the lowest.     

Effect of meridional wind on gap-leaping western boundary current

Using a 1.5-layer reduced-gravity nonlinear shallow-water equation model, we studied the effect of the meridional wind on the western boundary currents (WBC) at critical states with hysteresis courses. The results of the simulation indicate that the WBC is prone to penetrating into the gap under northerly winds, and its path is more difficult to alter due to the larger interval between the two critical transition curves (C ₁ P and C ₁ L). For southerly winds, the WBC is prone to leaping across the gap, and its path is easier to alter due to the smaller interval between the two critical transition curves. The simulation results also indicate that the meridional winds over the southern region of the gap are the dominant factor determining the formation of the WBC. The dynamic mechanism influencing the transport of WBC near the gap is both Ekman transport and the blocking of Ekman transport. Ekman transport induced by northerly winds may reduce the transport of the WBC, causing the β-effect to dominate the meridional advection (promoting the penetration). Southerly winds, however, may enhance the transport of the WBC, causing the meridional advection to dominate the β-effect (promoting the leaping state). These results explain some structural features of the Kuroshio at the Luzon Strait.     

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.