PLASMA CORTISOL AND GLUCOSE CONCENTRATIONS IN THE STRIPED MULLET (MUGIL CEPHALUS L.) SUBJECTED TO INTENSE HANDLING STRESS

The plasma cortisol and glucose concentrations were determined in mature female striped mullet (Mugil cephalus L . ) subjected to short term intense handling stress . The results indicated that plasma cortisol levels reached a peak 20 min after stress and declined gradually afterwards . The highest concentration of plasma glucose was observed 30 min after stress . The present study showed that the rise of plasma glucose was associated with the plasma cortisol levels .     

ANNUAL CHANGES IN PLASMA LEVELS OF CORTISOL AND SEX STEROID HORMONES IN MALE RAINBOW TROUT, ONCORHYNCHUS MYKISS

The profiles of cortisol, testosterone, ll-ketotestosterone and 17α， 20β- dihydroxy-4-pregnene-3-one in mule rainbow trout reared under constant water temperature and natural photoperiod were determined by radioimmunoassay. Gonads of male rainbow trout reached maturity when the fish were two years old. Changes in the plasma levels of both sex steroid hormones and cortisol were closely related to the GSI. Plasma levels of testosterone, 11-ketotestostemne and 17a, 2013-dihydroxy 4-pregnene-3-one showed a clear peak in the annual breeding season, when the GSI reached their maxima. Plasma cortisol levels also showed clearly seasonal changes in both two- and three-year-old fish. The results suggest that the elevated plasma levels of cortisol may not just be due to stresses during the breeding season but have certain physiological functions in the reproduction of rainbow trout.     

Effect of hypoxia on the blood of large yellow croaker (Pseudosciaena crocea)

Large areas of hypoxic water have recently been reported in the East China Sea. It is hypothesized that hypoxia may be partially responsible for the decline of some fish stocks. We evaluated the effect of hypoxia on large yellow croaker (Pseudosciaena crocea). The fish were exposed to three concentrations of dissolved oxygen (DO; 1.5 mg/L and 2.0 mg/L, and 6.5 mg/L control). We collected blood after 6, 12, 24, 48, and 96 h exposure. There was a significant increase in red blood count, hematocrit, hemoglobin concentration, and mean corpuscular hemoglobin in the group exposed to 1.5 mg/L DO after 6 h or 12 h, and a delayed increase (only elevated after 48 h and 96 h) in these indices in the group exposed to 2.0 mg/L DO. Plasma glucose concentrations increased significantly in both hypoxic groups after 24 h. Furthermore, plasma lactate and lactate dehydrogenase activity increased significantly after the first 6 h exposure in both hypoxic groups. Our results suggest that large yellow croakers could not maintain the aerobic pathway and instead use anaerobic metabolism for survival when DO levels fall below 2.0 mg/L. We conclude that the occurrence of hypoxia (<2 mg/L DO) in the East China Sea could cause metabolic stress for large yellow croakers and may be partially responsible for the population decline of this species.     

Low temperature stress on the hematological parameters and <Emphasis Type="Italic">HSP</Emphasis> gene expression in the turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>

To study the effect of low temperature stress on hematological parameters and HSP gene expression in the turbot (Scophthalmus maximus), water temperature was lowered rapidly from 18 to 1°C. During the cooling process, three individuals were removed from culture tanks at 18, 13, 8, 5, 3, and 1°C. Blood samples and tissues were taken from each individual, hematological indices and HSP gene expression in tissues were measured. The red blood cell count, white blood cell count, and hemoglobin concentration decreased significantly (P < 0.05) as temperature decreased. Enzyme activities of plasma alanine transaminase and creatine kinase increased as temperature decreased, whereas aspartic transaminase and γ-glutamyl transpeptidase activities displayed no obvious changes above 1°C and lactate dehydrogenase activity increased first and then decreased. Blood urea nitrogen and uric acid levels were highest at 8°C, and creatinine concentration was highest at 3°C. The concentrations of plasma cortisol, cholesterol, and triglyceride all increased significantly (P < 0.05) as temperature decreased. The serum glucose concentration increased first and then decreased to the initial level. The HSP70 mRNA expression showed various patterns in different tissues, whereas HSP90 mRNA expression showed the same tendency in all tissues. Overall, these results indicate that temperature decreases in the range of 8 to 5°C may induce a stress response in S. maximus and that temperature should be kept above 8°C in the aquaculture setting to avoid damage to the fish.     

低氧胁迫对珍珠龙胆石斑鱼氧化应激及能量代谢的影响

【目的】探究保活运输中低氧胁迫对珍珠龙胆石斑鱼(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂)氧化应激的影响。【方法】诱导体质量为(498.2±27.3)g的杂交石斑鱼进入休眠状态,并放置在溶解氧为(1.74±0.21)mg·L^?1的低氧环境下,低氧胁迫0、6、12、24、48、72 h后,取样测定血清应激指标,肌肉、肝脏、鳃组织氧化应激指标及肌肉、肝脏能量利用指标。【结果】低氧胁迫过程中,杂交石斑鱼血糖含量在胁迫6 h时最大,72 h时恢复胁迫前水平;皮质醇含量在胁迫过程均显著高于胁迫前;肌肉超氧化物歧化酶(SOD)活性随胁迫时间的增加而上升,肝脏、鳃SOD活性先上升后下降;肌肉、肝脏过氧化氢酶(CAT)活性呈先升后降的趋势,鳃CAT活性在胁迫12、72 h显著低于胁迫前;肌肉、肝脏丙二醛(MDA)含量在胁迫过程中均增加,鳃MDA含量先降后升;肌肉谷胱甘肽还原酶(GSH)含量在胁迫12 h达到最大值,肝脏GSH含量呈先上升后下降的趋势,鳃中GSH含量在12 h时显著高于胁迫前,之后逐渐升高;总抗氧化能力(T-AOC)在肌肉、肝脏、鳃中均呈上升的趋势;肌肉和肝脏中糖原的含量显著减少,肌肉乳酸脱氢酶(LDH)活性显著增加,肝脏LDH先降后升。显著性水平均为0.05。【结论】低氧胁迫会刺激珍珠龙胆石斑鱼产生应激反应,影响体内正常的氧化应激系统,并对不同组织造成不同程度的氧化损伤。     

Mechanical Stress Induces Neuroendocrine and Immune Responses of Sea Cucumber(Apostichopus japonicus)

Grading procedure in routine sea cucumber hatchery production is thought to affect juvenile sea cucumber immunological response. The present study investigated the impact of a 3-min mechanical perturbation mimicking the grading procedure on neuroendocrine and immune parameters of the sea cucumber Apostichopus japonicus. During the application of stress, concentrations of noradrenaline and dopamine in coelomic fluid increased significantly, indicating that the mechanical perturbation resulted in a transient state of stress in sea cucumbers. Coelomocytes concentration in coelomic fluid increased transiently after the beginning of stressing, and reached the maximum in 1 h. Whereas, coelomocytes phagocytosis at 3 min, superoxide anion production from 3 min to 0.5 h, acid phosphatase activity at 0.5 h, and phenoloxidase activity from 3 min to 0.5 h were all significantly down-regulated. All of the immune parameters recovered to baseline levels after the experiment was conducted for 8 h, and an immunostimulation occurred after the stress considering the phagocytosis and acid phosphatase activity. The results suggested that, as in other marine invertebrates, neuroendocrine/immune connections exist in sea cucumber A. japonicus. Mechanical stress can elicit a profound influence on sea cucumber neuroendocrine system. Neuroendocrine messengers act in turn to modulate the immunity functions. Therefore, these effects should be considered for developing better husbandry procedures.     

Acclimation-dependent expression of heat shock protein 70 in Pacific abalone (Haliotis discus hannai Ino) and its acute response to thermal exposure

Heat shock protein 70 (Hsp70) is one important member of heat shock protein (Hsp) family that is responsible for various stresses, especially thermal stress. Here we examined the response of Hsp70 gene to both chronic and acute thermal exposure in Pacific abalone (Haliotis discus hannai Ino). For the chronic exposure, abalones were maintained at 8, 12, 20, and 30°C for four months and their mRNA levels were measured. The highest mRNA level of Hsp70 gene relative to actin gene was detected in the 30°C-acclimated group, followed by the 8°C-acclimated group and then the 12°C- and 20°C-acclimated groups. After the long-term acclimation, gills from each of the above acclimation groups were dissected and exposed to different temperatures between 8°C and 38°C for 30 min. Hsp70 expression in gills acclimated to different temperatures responded differentially to the same temperature exposure. The incubation temperature that induced maximum Hsp70 mRNA expression was higher in the higher temperature acclimation groups than lower temperature groups. Pacific abalones could alter the expression pattern of Hsp70 gene according to environmental thermal conditions, through which they deal with the stress of thermal variations.     

急冷与空气暴露联合胁迫对凡纳滨对虾生理代谢影响

【目的】探明无水保活流通过程中凡纳滨对虾(Litopenaeus vannamei)响应急冷(acute cold,AC)与空气暴露(waterless duration,WD)联合胁迫的生理代谢调节规律。【方法】研究模拟产业运输实际,分析联合胁迫(AC+WD)进程中凡纳滨对虾血液和肝胰腺典型生化指标,以及肝胰腺组织病理变化。【结果】AC胁迫后,凡纳滨对虾生理应激指标血清皮质醇含量由正常组(normal control,NC)的(326.90±4.61)pg/mL显著增加到AC组的(427.16±7.92)pg/mL(P<0.05),AC+WD组则随时间延长而逐渐降低。血清葡萄糖含量在联合胁迫6 h后达到最大值(26.31±1.05)mg/dL,乳酸在9 h达到峰值(7.90±0.11)mmol/L。AC及AC+WD组中血蓝蛋白、尿素氮和肌酐含量较NC组均发生不同程度的增加,而总蛋白含量先升后降,总胆固醇和甘油三酯含量呈胁迫时间依赖性下降。血清谷草转氨酶、谷丙转氨酶、酸性和碱性磷酸酶活力呈上升趋势,并在9 h达到峰值。以上生化指标在胁迫复苏组有所恢复,表明AC+WD双重胁迫引起凡纳滨对虾物质代谢紊乱,复苏后机体生理应激和代谢系统发生一系列代偿性调节,且胁迫进程中肝胰腺组织病理损伤特征进一步解释了凡纳滨对虾响应环境联合胁迫的结构基础。【结论】无水保活流通过程中双重胁迫时间依赖性改变凡纳滨对虾生理应激水平和物质代谢,在9 h时可明显造成肝胰腺组织结构损伤,进而影响机体存活。     

Study on application of polyenzyme method to offal of Harengula zunasi

The new polyenzyme method for making gravy fromHarengula zunasi offal involves protein enzymolysis with flavorase after proper alkaline and neutral protease levels were established by orthogonal trials to select the best hydrolytic conditions for processing offal with alkaline and neutral protease. The conditions for the polyenzyme method were pH of 7.0, temperature of 50°C, alkaline and neutral protease concentration of 1.5% respectively, hydrolysis time of 120 min, and flavorase concentration of 2.0%, for 60 min. The new gravy-making technology yields a nutritious and delicious gravy containing 40.3% of total essential amino acids, with delicious amino acids Glu, Asp, Gly, Ala, Pro and Ser comprising 49.5%, total and amino nitrogen being respectively 1.9 and 1.1 g/100 g (amino acid nitrogen being 61.0% of total nitrogen), The polyenzyme method was used to make 14.8% protein gravy fromHarengula zunasi offal. In addition, inorganic elements, the phosphorus content is the highest. This project was aided financially by the Guangdong Science and Technology Office (No. 2KM06002S).     

Physiological responses of Chinese longsnout catfish to water temperature

We evaluated the effect of water temperature on the growth and physiology of the Chinese longsnout catfish (Leiocassis longirostris Günther). The fish were reared at four temperatures (20, 25, 30, and 35°C) and sampled on days 7, 20, and 30. We measured plasma levels of insulin, free thyroxine (FT4), free 3,5,3′-triiodothyronine (FT3), lysozyme and leukocyte phagocytic activity. The optimum water temperature for growth was 27.7°C. The plasma levels of insulin and FT4 declined significantly (P<0.05) on day 30 at temperatures above 20°C. Lysozyme activity was significantly (P<0.05) lower at 25°C than at other temperatures. We conclude that final weight, insulin, FT4, and lysozyme were significantly affected by water temperature.     

Optimization of dilute acid hydrolysis of Enteromorpha

Acid hydrolysis is a simple and direct way to hydrolyze polysaccharides in biomass into fermentable sugars. To produce fermentable sugars effectively and economically for fuel ethanol, we have investigated the hydrolysis of Enteromorpha using acids that are typically used to hydrolyze biomass:H2SO4, HCl, H3PO4 and C4H4O4 (maleic acid). 5%(w/w) Enteromorpha biomass was treated for different times (30, 60, and 90 min) and with different acid concentrations (0.6, 1.0, 1.4, 1.8, and 2.2%, w/w) at 121°C. H2SO4 was the most effective acid in this experiment. We then analyzed the hydrolysis process in H2SO4 in detail using high performance liquid chromatography. At a sulfuric acid concentration of 1.8% and treatment time of 60 min, the yield of ethanol fermentable sugars (glucose and xylose) was high, (230.5 mg/g dry biomass, comprising 175.2 mg/g glucose and 55.3 mg/g xylose), with 48.6% of total reducing sugars being ethanol fermentable. Therefore, Enteromorpha could be a good candidate for production of fuel ethanol. In future work, the effects of temperature and biomass concentration on hydrolysis, and also the fermentation of the hydrolysates to ethanol fuel should be focused on.     

Protective effects of Ecklonia cava extract on the toxicity and oxidative stress induced by hair dye in in-vitro and in-vivo models

Oxidative hair dyes containing p-phenylenediamine(PPD) are reported to induce an allergic reaction by promoting oxidative stress when absorbed through the skin. Despite the associated risk, these hair dyes remain popular owing to their convenience and sharpness of color. This makes it important to minimize the cytotoxicity and oxidative stress induced by PPD-containing hair dyes. Ecklonia cava extract has been evaluated in different studies for its protective effects against external stress in fibroblasts and keratinocytes. Our study was aimed at using in-vitro and in-vivo models to investigate the extract's effects on cytotoxicity of and oxidative stress induced by PPD-containing hair dyes. Analysis of CIEL*a*b* Color space was first used to determine the range of E. cava extract that would not interfere with the coloring ability of the dye upon addition. Subsequently, the set ranges of E. cava extract(5% and 7%) were added to the hair dye and their toxicity assessed by evaluating the viability of fibroblasts and keratinocytes. The effects on developmental phenotypes and induction of oxidative stress by hair dye were evaluated and compared with those of hair dyes containing different contents of E. cava extract using an in-vivo zebrafish model. Our results showed that E. cava extract in hair dye could significantly decrease the cytotoxicity and levels of oxidative stress caused by hair dyes containing PPD in both in-vitro and in-vivo models.These results suggest that the addition of 7% E. cava extract to 250 μg/mL hair dye does not interfere with the coloring ability of the dye while showing significant protective effects against the hair dye. The study proposes that the use of E. cava extract as an adduct to hair dyes containing PPD reduces the cytotoxicity and oxidative stress induced by these hair dyes.     

Heat-shock protein 70 expression in shrimp Fenneropenaeus chinensis during thermal and immune-challenged stress

Using western immunoblotting we obtained heat-shock protein 70 (HSP70) induction data and distribution in different tissues from shrimpFenneropenaeus chinensis during thermal and immune-challenged stresses. This is probably the first report of the effects of various stressors on the expression of HSP70 in shrimp. HSP70 was prominently induced in hepatopancreas and gills, but not in muscle, eyestalk and hemolymph, when the shrimp were exposed to heat shock andVibrio anguillavium-challenged stresses. Cold shock and WSSV treatment had no significant effects on the levels of HSP70 expression in all tissues examined. HSP70 induction was greatest after 2 h exposure to heat shock stress, which was elevated after acute heat shock exposure of 10°C above ambient temperature. Supported by the National Key Fundamental Research Program “Study of the Major Diseases and Resistance Mechanism of Mariculture Organisms”, NSFC, No. 30140017, and the international cooperative program “Immunaqua” by the European Commission, No. ICA4-CT-2001-10023.     

Variations of antioxidant enzyme activity and malondialdehyde content in nemertean Cephalothrix hongkongiensis after exposure to heavy metals

The antioxidant enzyme activity and malondialdehyde (MDA) content of Cephalothrix hongkongiensis were studied to assess variations in the biochemical/physiological parameters of nemerteans under heavy metal stress. Worms were exposed to copper, zinc and cadmium solutions at different concentrations, and the activity of three antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX), and MDA content were measured. The results show that the activity of each enzyme changed immediately after exposure to heavy metals. CAT was invariably inhibited throughout the experimental period, while the SOD activity was significantly elevated by exposure to Cu²⁺ for 48 h, but then decreased. SOD was inhibited by Zn²⁺during the first 12 h of exposure, but activated when exposed for longer periods. Under Cd²⁺ stress, SOD activity decreased within 72 h. GPX activity varied greatly, being significantly increased by both Cu²⁺ and Zn²⁺, but significantly inhibited by Cd²⁺ in the first 12–24 h after exposure. MDA content increased on Cu²⁺ exposure, but normally decreased on Zn²⁺ exposure. MDA content followed an increase-decrease-increase pattern under Cd²⁺ stress. In conclusion, the antioxidant system of this nemertean is sensitive to heavy metals, and its CAT activity may be a potential biomarker for monitoring heavy metal levels in the environment.     

A successful microbound diet for the larval culture of Chinese shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselmis chuii, rotifer (Brachionus plicatilis) and Artemia sp. Larvae of 0 to 13d post-hatch (dph) were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L⁻¹ in tanks, each containing 50 L sterilized seawater with salinity 30–32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13 dph, which was not significantly different from that of larvae fed on live foods (63.55% ). The body length and development index (DI) of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13dph (P<0.05). The mean dry weight loss of the microbound diet was 9.2% after 90min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 5223% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

Numerical study on the bifurcation of the North Equatorial Current

A 1.5-layer reduced-gravity model forced by wind stress is used to study the bifurcations of the North Equatorial Current(NEC).The authors found that after removing the Ekman drift,the modelled circulations can serve well as a proxy of the SODA circulations on the σθ=25.0 kg m～-3 potential density surface based on available long-term reanalysis wind stress data.The modelled results show that the location of the western boundary bifurcation of the NEC depends on both zonal averaged and local zero wind stress curl latitude.The effects of the anomalous wind stress curl added in different areas are also investigated and it is found that they can change the strength of the Mindanao Eddy(ME),and then influence the interior pathway.     

Effect of dissolved oxygen on swimming ability and physiological response to swimming fatigue of whiteleg shrimp (Litopenaeus vannamei)

The swimming endurance of whiteleg shrimp(Litopenaeus vannamei, 87.66 mm ± 0.25 mm, 7.73 g ± 0.06 g) was examined at various concentrations of dissolved oxygen(DO, 1.9, 3.8, 6.8 and 13.6 mg L-1) in a swimming channel against one of the five flow velocities(v1, v2, v3, v4 and v5). Metabolite contents in the plasma, hepatopancreas and pleopods muscle of the shrimp were quantified before and after swimming fatigue. The results revealed that the swimming speed and DO concentration were significant factors that affected the swimming endurance of L. vannamei. The relationship between swimming endurance and swimming speed at various DO concentrations can be described by the power model(ν·tb = a). The relationship between DO concentration(mg L-1) and the swimming ability index(SA∫ 9000I), defined as SAI =vdt( cm), can be described as SAI = 27.947 DO0.137(R2 = 0.9312). The 0level of DO concentration directly affected the physiology of shrimp, and exposure to low concentrations of DO led to the increases in lactate and energetic substrate content in the shrimp. In responding to the low DO concentration at 1.9 mg L-1 and the swimming stress, L. vannamei exhibited a mix of aerobic and anaerobic metabolism to satisfy the energetic demand, mainly characterized by the utilization of total protein and glycogen and the production of lactate and glucose. Fatigue from swimming led to severe loss of plasma triglyceride at v1, v2, and v3 with 1.9 mg L-1 DO, and at v-11 with 3.8, 6.8 and 13.6 mg L DO, whereas the plasma glucose content increased significantly at v3, v4 and v5 with 3.8 and 6.8 mg L-1 DO, and at v5 with 13.6 mg L-1 DO. The plasma total protein and hepatopancreas glycogen were highly depleted in shrimp by swimming fatigue at various DO concentrations, whereas the plasma lactate accumulated at high levels after swimming fatigue at different velocities. These results were of particular value to understanding the locomotory ability of whiteleg shrimp and its physiological changes, further contributing to the improvement of capture and rearing technique.     

CO2协同低温有水对卵形鲳鲹麻醉保活效果的影响

【目的】探究二氧化碳CO₂麻醉协同低温对卵形鲳鲹有水保活的影响,延长卵形鲳鲹(Trachinotus ovatus)的保活时间。【方法】研究保活过程中CO₂质量浓度、麻醉时间、麻醉温度和鱼质量水比对卵形鲳鲹有水保活时长的影响;在最佳麻醉保活条件下,测定其血清生化指标、肌肉及肝脏氧化应激指标及代谢指标。【结果】卵形鲳鲹在16℃、100 mg/L CO₂水溶液中麻醉3 min其麻醉效果最佳。在15℃低温下进行有水保活,鱼水质量比为2∶1,存活时间最长,为(359±2)min,此条件下复苏率达100%。经过CO₂麻醉协同低温有水保活后的卵形鲳鲹血清谷草转氨酶(GOT)活性显著升高(P<0.05)、尿素氮(BUN)和甘油三酯(TG)含量上升,血糖含量下降。肌肉中的乳酸脱氢酶(LDH)活性显著上升(P<0.05),乳酸(LD)和糖原含量上升,过氧化氢酶(CAT)活性和丙二醛(MDA)含量下降;肝脏中的LDH活性显著上升(P<0.05)、CAT活性和LD的含量上升,MDA和糖原的含量下降。【结论】CO₂协同低温有水对卵形鲳鲹有较好的麻醉效果,有助于保活运输。在保活过程中卵形鲳鲹通过提升能量代谢水平和抗氧化能力应答环境应激,在一定程度上影响卵形鲳鲹保活过程的存活时间。     

EFFECTS OF FISH OIL,DHA OIL AND LECITIHIN IN MICROPARTICULATE DIETS ON STRESS TOLERANCE OF LARVAL GILTHEAD SEABREAM（SPARUS AURATA）

The effects of natural fish oil,DHA oil and soybean lecithin in microparticulate diets on stress tolerance of larval gilthead seabream(Sparus aurata)were investigated after 15 days feeding trials.The tolerance of larval gilthead seabream to various stress factors such as exposure to air(lack of dissolved oxygen),changes in water temperature(low)and salinity(high) were determined.This study showed that microparticulate diet with natural fish oil and soybean lecithin was the most effective for in-creasing the tolerance of larval gilthead seabream to various stresses,and that microparticulate diet with natural fish oil and palmitic acid(16:0)was more effective than microparticulate diet with DHA oil and soybean lecithin.     

Chromosome behavior of heat shock induced triploid in Fenneropenaeus chinensis

Triploidy was induced in Chinese shrimpFenneropenaeus chinensis by 30±0.5°C heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0°C. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs. Contribution No. 4407 from the Institute of Oceanology, Chinese Academy of Sciences. This work was supported by Project under Major State Basic Development Program (G1999012009) and Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX 2-211)