Influence of several non-nutrient additives on nonspecific immunity and growth of juvenile turbot, Scophthalmus maximus L.

The effects of three non-nutrient additives on nonspecific immunity and growth of juvenile turbot (Scophthalmus maximus L.) were studied in this feeding experiment. The five treatments are basal diet alone, basal diets containing three different additives [0.4 g kg(-1) of xylo-oligosaccharides (XOS), 1.3 g kg (-1) of yeast cell wall and 0.8 g kg (-1) of bile acids] individually or in combination. Two hundred and twenty-five turbots (average initial weight 151.3 +/- 11.3 g) were randomly allotted in five treatments with three replicates within each treatment in a 72-day period. Comparing with basal diet group, activities of C3, C4, phagocyte, lysozyme, specific growth rate and feed conversion rate in yeast cell wall, XOS and the combined groups was enhanced significantly (P < 0.05); however, these parameters in bile acid groups were increased slightly (P > 0.05) except for phagocyte (P < 0.05); superoxide dismutase activity in additive groups was not significantly increased (P > 0.05) except for the combined group (P < 0.05). In conclusion, supplementation of yeast cell wall and XOS enhanced the nonspecific immunity of juvenile turbot. Synergistic or additive effect of the three additives was not observed.

Domain analysis of the Edwardsiella tarda ferric uptake regulator

Recent studies have shown that the ferric uptake regulator (Fur) of Edwardsiella tarda (Fur(Et)) shares high sequence identity with the Escherichia coli Fur (Fur(Ec)) at the N-terminal DNA-binding region. In the present study, the functional importance of the C-terminal region of Fur(Et) was investigated. It was found that Fur(Et) bearing deletion of the C-terminal 12 residues still possesses most of the repressor activity, whereas Fur(Et) bearing deletions of the C-terminal 16 and more than 16 residues are severely affected in activity. Domain swapping analyses indicated that the chimeric Fur proteins (Et75Ec73 and Et75Vh74) consisting of the N-terminal 1-75 region of Fur(Et) fused to the C-terminal 76-148 region of Fur(Ec) and the C-terminal 76-149 region of the Vibrio harveyi Fur (Fur(Vh)), respectively, are fully active. C92 of Fur(Ec) and C137 of Fur(Vh), which are functionally essential in Fur(Ec) and Fur(Vh), respectively, are also essential in Et75Ec73 and Et75074, respectively. Further study identified an artificial Fur protein, EtMF54, which is composed of the N-terminal 49 residues of Fur(Et) and five artificial residues. Compared to Fur(Et), EtMF54 possesses partial Fur activity that is iron-dependent. These results (I) indicate that there exist certain functional/structural compatibilities among Fur(Et), Fur(Ec), and Fur(Vh) at the C-terminal region; (ii) provide insights to the potential location of the regulatory ion-binding site of Fur(Et).

Cys-92, Cys-95, and the C-terminal 12 residues of the Vibrio harveyi ferric uptake regulator (Fur) are functionally inessential

Ferric uptake regulator (Fur) is a global regulator involved in multiple aspects of bacterial life. The gene encoding the Vibrio harveyi Fur (Fur(vh)) was cloned from a pathogenic V. harveyi strain isolated from diseased fish. Furvh shares 77% overall sequence identity with the Escherichia coli Fur (Fur(Ec)) and could complement a mutant of Fur(Ec). Like Fur(Ec), Fur(Vh), possesses two cysteine residues at positions 92 and 95, yet unlike Fur(Ec), in which these cysteine residues constitute part of the metal ion coordination site and hence are vital to the repressor activity, C92 and C95 of Fur(Vh) proved to be functionally inessential. Further study identified a Vibrio Fur signature sequence, which is preserved in all the ten Vibrio Fur proteins that have been discovered to date but in none of the non-vibrio Fur proteins. Site-directed and random mutation analyses of the signature residues, the cysteine residues, and seven highly charged amino acid residues indicated that D9, H32, C137, and K138 of Fur(vh) are functionally important but D9, C137, and K138 can be replaced by more than one functional substitutes. Systematic deletion analysis demonstrated that the C-terminal 12 residues of Fur(Vh) are functionally inessential. These results (i) indicated that the activation mechanism, or certain aspects of which, of Fur(Vh) is possibly different from that of Fur(Ec); and (ii) suggested that it is not very likely that the C-terminal 12 residues play any significant role in the activation or stability of Fur(Vh); and (iii) provided insights into the potential function of the local structure involving C137 and K138.

Molecular analysis of the fur (ferric uptake regulator) gene of a pathogenic Edwardsiella tarda strain

The gene encoding the Edwardsiella tarda ferric uptake regulator (Fur(Et)) was cloned from a pathogenic E. tarda strain isolated from diseased fish. Fur(Et) shares 90% overall sequence identity with the Escherichia coli Fur (Fur(Ec)) and was able to complement the mutant phenotype of a fur(Ec)-defective E. coli strain. Mutational analysis indicated that C92S and C95S mutations inactivated Fur(Et) whereas E112K mutation resulted in a superactive Fur(Et) variant. Fur(Et) negatively regulated its own expression; interruption of this regulation impaired bacterial growth, altered the production of certain outer membrane proteins, and attenuated bacterial virulence.

Growth of Gracilaria lemaneiformis under different cultivation conditions and its effects on nutrient removal in Chinese coastal waters

Gracilaria lemaneiformis (Bory) Daws has been extensively cultivated as a source of commercial agar and the ecomaterials in Shenao Bay, Guangdong Province, Jiaozhou Bay, Shandong Province and other waters in China. This paper examines the in situ suspended farming of G. lemaneiformis using raft cultivation under different conditions and its effects on nutrient removal in the laboratory. The results showed that cultivated Gracilaria grew well in both Shenao Bay and Jiaozhou Bay. The biomass of Gracilaria increased from 50 to 775 g m(-1) (fresh weight) during 28 days, with special growth rate (SPG) 13.9% d(-1) under horizontal cultivation in Jiaozhou Bay. Light, temperature, nutrient supply, as well as cultivation treatments such as initial density, and depth of suspension seaweed were important to the growth of Gracilaria. The highest biomass production was observed in the horizontal culture condition (0.0 m) and 0.5-1.5 m deep layer in Jiaozhou Bay. However, the highest growth rate in Shenao Bay appeared under the lowest initial stocking density treatment. In the laboratory, the aquarium experiments (fish and seaweed culture systems) demonstrated that Gracilaria was able to remove inorganic nutrients effectively. The concentration of NH4+-N decreased by 85.53% and 69.45%, and the concentration of PO4-P decreased 65.97% and 26.74% in aquaria with Gracilaria after 23 days and 40 days, respectively. The results indicate that Gracilaria has the potential to remove excess nutrient from coastal areas, and the large-scale cultivation of G. lemaneiformis could be effective to control eutrophication in Chinese coastal waters. (c) 2005 Elsevier B.V. All rights reserved.

Numerical investigation of nutrient limitations in the Bohai Sea

River discharges are the important freshwater and nutrient sources for Bohai Sea (BS), and have a profound impact on the local marine environment. In this paper, the annual cycles of nutrient and phytoplankton dynamics in 1980s were reproduced using a coupled biogeochemical-physical model. Based on the validated simulations, the nutrient limitation characters were further investigated by running the model with the riverine nutrient altered, first enriching nitrogen and then phosphorus. It was found that although the riverine N:P ratios in Yellow and Haihe Rivers were much higher than the Redfield number, the nitrogen enrichment was still able to enhance the algae bloom in Laizhou and Bohai Bays. On the other hand, the response of algae growth to phosphorus enrichment was not thus obvious, which suggests that the local phytoplankton dynamics was characterized by the nitrogen limitation. Simulations also show that the nitrogen enrichment is generally accompanied by the phosphorus consumption, so a shift from nitrogen limitation to phosphorus limitation may occur if such a trend continues. (C) 2010 Elsevier Ltd. All rights reserved.

Relationships between nitrogen and phosphorus forms and ratios and the development of dinoflagellate blooms in the East China Sea

During late spring and early summer of 2005, large-scale (> 15 000 km(2)), mixed dinoflagellate blooms developed along the the coast of the East China Sea. Karenia mikimotoi was the dominant harmful algal bloom species in the first stage of the bloom (late May) and was succeeded by Prorocentrum donghaiense approximately 2 wk later. Samples were collected from different stations along both north-south and west-east transects, from the Changjiang River estuary to the south Zhejiang coast, during 3 cruises of the Chinese Ecology and Oceanography of Harmful Algal Blooms Program, before and during the bloom progression. Nitrogen isotope tracer techniques were used to measure rates of NO3-, NH4+, urea, and glycine uptake during the blooms. High inorganic nitrogen (N), but low phosphorus (P) loading from the Changjiang River led to high dissolved inorganic N:dissolved inorganic P ratios in the sampling area and indicate the development of P limitation. The rates of N-15-uptake experiments enriched with PO43- were enhanced compared to unamended samples, suggesting P limitation of the N-uptake rates. The bloom progression was related to the change in availability of both organic and inorganic N and P. Reduced N forms, especially NH4+, were preferentially taken up during the blooms, but different bloom species had different rates of uptake of organic N substrates. K mikimotoi had higher rates of urea uptake, while P. donghaiense had higher rates of glycine uptake. Changes in the availability of reduced N and the ratios of N:P in inorganic and organic forms were suggested to be important in the bloom succession. Nutrient ratios and specific uptake rates of urea were similar when compared to analogous blooms on the West Florida Shelf.

Nutrient characteristics in the Yangtze River Estuary and the adjacent East China Sea before and after impoundment of the Three Gorges Dam

From November 2002 to 2006, five cruises were undertaken in the Yangtze River Estuary and the adjacent East China Sea to compare the nutrient concentrations, ratios and potential nutrient limitation of phytoplankton growth before and after impoundment (June 2003) of the Three Gorges Dam (TGD). Concentrations of dissolved inorganic nitrogen (DIN), soluble reactive phosphorus (SRP) and total nitrogen (TN) exhibited an increasing trend from 2002 to 2006. In contrast, total phosphorus (TP) concentration exhibited a decreasing trend. The mean concentrations of DIN, SRP, and TN in the total study area increased from 21.4 mu M, 0.9 mu M, and 41.8 mu M in 2002 to 37.5 mu M, 1.3 mu M. and 82.2 mu M in 2006, respectively. while TP decreased from 2.1 mu M to 1.7 mu M. The concentration of dissolved reactive silica (DRSi) had no major fluctuations and the differences were not significant. The mean concentration of DRSi in the total study area ranged from 52.5 to 92.3 mu M. The Si:N ratio decreased significantly from 2.7 in 2002 to 1.3 in 2006, while TN: TP ratio increased from 22.1 to 80.3. The area of potential P limitation of phytoplankton growth expanded after 2003 and potential Si limitation appeared in 2005 and 2006. Potential P limitation mainly occurred in an area of salinity less than 30 after 2003, while potential Si limitation occurred where the salinity was greater than 30. By comparison with historical data, the concentrations of nitrate and SRP in this upper estuary during November 1980-2006 increased obviously after impoundment of TGD but DRSi decreased. Meanwhile, the ratios of N:P, Si:N and Si:P decreased obviously. (c) 2009 Elsevier B.V. All rights reserved.

Estuarine nutrient loading affects phytoplankton growth and microzooplankton grazing at two contrasting sites in Hong Kong coastal waters

To investigate the effects of enhanced nutrient loading in estuarine waters on phytoplankton growth and microzooplankton grazing, we conducted monthly dilution experiments at 2 stations in Hong Kong coastal waters with contrasting trophic conditions. The western estuarine station (WE) near the Pearl River estuary is strongly influenced by freshwater discharge, while the eastern oceanic station (EO) is mostly affected by the South China Sea. Growth rates of phytoplankton were often limited by nutrients at EO, while nutrient limitation of phytoplankton growth seldom Occurred at WE due to the high level of nutrients delivered by the Pearl River, especially in the summer rainy season. Higher chlorophyll a, microzooplankton biomass, phytoplankton growth and microzooplankton grazing rates were found at WE than at EO. However, the increase in chlorophyll greatly exceeded the increase in phytoplankton growth rate, reflecting different response relationships to nutrient availability. Strong seasonality was observed at both stations, with temperature being an important factor affecting both phytoplankton growth and microzooplankton grazing rates. Picophytoplankton, especially Synechococcus, also exhibited great seasonality at EO, with summer abundances being 2 or 3 orders of magnitude higher than those during winter, Our results confirm that in eutrophic coastal environments, microzooplankton grazing is a dominant loss pathway for phytoplankton, accounting for the utilization of >50%, of primary production on average.

Nutrient Dynamics in the Upwelling Area of Changjiang (Yangtze River) Estuary

Nutrient dynamics and its influence on the distribution of chlorophyll-a in the upwelling area of the Changjiang (Yangtze) River estuary were investigated in the spring (May) and summer (August) of 2004. In the spring, upwelling was apparent in the region of 122 degrees 20'-123 degrees 00' E, 31 degrees 00'-32 degrees 00' N and was associated with low temperature (16-21 degrees C), high salinity (24-33 practical salinity units [psu]), and low dissolved oxygen (2.5-6.0 mg L-1) in the upper 10 m of the water column. The spring upwelling increased the mixed-layer phosphate, nitrate, and silicate concentrations to roughly 1, 15, and 15 mu mol L-1, respectively, and improved the light transparency in the euphotic zone. This improvement in phytoplankton growing conditions was followed by an increase in chlorophyll-a concentrations. The summer upwelling was weaker and occurred over a smaller geographical area (122 degrees 20'-123 degrees 00' E, 31 degrees 15'-31 degrees 50' N). Strongly influenced by turbid Changjiang diluted water (CDW), it had little impact on the upper 10 m of the water column but instead increased nutrient concentrations at greater depths. The high concentration of particulates in the CDW reduced light transmission in the upper 10 m and, hence, limited phytoplankton growth throughout the water column. Chlorophyll-a concentrations in the summer upwelling area were roughly an order of magnitude lower than in the spring. Water clarity, as influenced by the CDW, appears to be the principal factor limiting the impact of upwelling on phytoplankton biomass in this area.

Biochemical compositions of two dominant bloom- forming species isolated from the Yangtze River Estuary in response to different nutrient conditions

Variations of cellular total lipid, total carbohydrate and total protein content of two dominant bloom-forming species (Skeletonema costatum and Prorocentrum donghaiense) isolated from the Yangtze River Estuary were examined under six different nutrient conditions in batch cultures. Daily samples were collected to estimate the cell growth, nutrient concentration and three biochemical compositions content during 7 days for S. costatum and the same sampling procedure was done every other day during 10 days for P. donghaiense. Results showed that for S. costatum, cellular total lipid content increased under phosphorus (P) limitation, but not for nitrogen (N) limitation; cellular carbohydrate were accumulated under both N and P limitation: cellular total protein content of low nutrient concentration treatments were significantly lower than that of high nutrient concentration treatments. For P. donghaiense, both cellular total lipid content and total carbohydrate content were greatly elevated as a result of N and P exhaustion, but cellular total protein content had no significant changes under nutrient limitation. In addition, the capability of accumulation of three biochemical constituents of P. donghaiense was much stronger than that of S. costatum. Pearson correlation showed that for both species, the biochemical composition of three constituents (lipid, carbohydrate and protein) had no significant relationship with extracellular N concentration, but had positive correlation with extracellular and intracellular P concentration. The capability of two species to accumulate cellular total lipid and carbohydrate under nutrient limitation may help them accommodate the fluctuating nutrient condition of the Yangtze River Estuary. The different responses of two species of cellular biochemical compositions content under different nutrient conditions may provide some evidence to explain the temporal characteristic of blooms Caused by two species in the Yangtze River Estuary. (C) 2008 Elsevier B.V. All rights reserved.

Causes and consequences of changes in nutrient structure in the Jiaozhou Bay

Concentrations and ratios of nutrients in Jiaozhou Bay, China, have changed much in the past decades, with trends indicating an increase in nitrogen and a decrease in silicate. Statistical analysis has shown that the long-term variations of nutrients are associated with agricultural activities, precipitation, and anthropogenic factors. Stoichiometric calculations indicate that the nutrient structure has become more and more unbalanced. There has been almost no possibility for nitrogen limitation since the 1980s, the probability of P limitation has increased, and the probability of Si limitation has also increased markedly from the 1980s to the 1990s. As a consequence of changes in nutrient structure, a decrease in the abundance of net phytoplankton was evident, whereas total chlorophyll a levels have remained roughly unchanged at around 3.55 mu g/L. Thus, it is likely that smaller species have taken the niche vacated by the larger species. Changes in phytoplankton size and species composition may ultimately lead to various functional and structural changes at the system level.

Evolution of nutrient structure and phytoplankton composition in the Jiaozhou Bay ecosystem

The inventories of nutrients in the surface water and large phytoplankton( > 69 pm) were analyzed from the data set of JERS ecological database about a typical coastal waters, the Jiaozhou Bay, China, from 1960s for N, P and from 1980s; for Si. By examining long-term changes of nutrient concentration, calculating stoichiometric balance, and comparing diatom composition, Si limitation of diatom production was found to be more possible. The possibility of Si limitation was from 37% in 1980s to 50% in 1990s. Jiaozhou Bay ecosystem is becoming serious eutrophication, with notable increase of NO2-N, NO3-N and NH4-N from 0.1417 mumol/L, 0.5414 mumol/L, 1.7222 mumol/L in 1960s to 0.9551 mumol/L, 3.001 mumol/L, 8.0359 mumol/L in late 1990s respectively and prominent decrease of Si from 4.2614 mumol/L in 1980s to 1.5861 mumol/L in late 1990s; the nutrient structure is controlled by nitrogen; the main limiting nutrient is probably silicon; because of the Si limitation the phytoplankton community structure has changed drastically.

Oceanic iron fertilization: one of strategies for sequestration atmospheric CO2

Carbon cycle is connected with the most important environmental issue of Global Change. As one of the major carbon reservoirs, oceans play an important part in the carbon cycle. In recent years, iron seems to give us a good news that oceanic iron fertilization could stimulate biological productivity as CO2 sink of human-produced CO2. Oceanic iron fertilization experiments have verified that adding iron into high nutrient low chlorophyll (HNLC) seawaters can increase phytoplankton production and export organic carbon, and hence increase carbon sink of anthropogenic CO2, to reduce global warming. In sixty days, the export organic carbon could reach 10 000 times for adding iron by model prediction and in situ experiment, i.e. the atmospheric CO2 uptake and inorganic carbon drawdown in upper seawaters also have the same magnitude. Therefore, oceanic iron fertilization is one of the strategies for increasing carbon sink of anthropogenic CO2. The paper is focused on the iron fertilization, especially in situ ocean iron experiments in order that the future research is more efficient.

Historical changes in nutrient structure and its influences on phytoplantkon composition in Jiaozhou Bay

Due to the influence of human activities, nutrient concentrations, nutrient ratios and phytoplankton composition have notably changed in Jiaozhou Bay, China since the 1960s. From the 1960s to the 1990s, nutrient concentrations have increased 1.4 times for PO4-P, 4.3 times for NO3-N, 4.1 times for NH4-N and 3.9 times for DIN. The atomic ratio of DIN:PO4-P increased very rapidly from 15.9 +/- 6.3 for the 1960s, to 37.8 +/- 22.9 for the 1990s. SiO3-Si concentration has remained at a very low level from the 1980s to the 1990s. The high ratio of DIN: PO4-P and low ratios of SiO3-Si:PO4-P (7.6 +/- 8.9) and SiO3-Si:DIN (0.19 +/- 0.15) showed the nutrient structure of Jiaozhou Bay has changed from more balanced to unbalanced during the last 40 years. The possibility that DIN and/or PO4-P as limiting factors of Jiaozhou Bay phytoplankton has been lessened or eliminated and that of SiO3-Si limiting has been increased. The changes in nutrient structure may have led to the decrease of large diatoms and a shift of phytoplankton species composition. It is likely that there is a trend from large diatoms to smaller cells in Jiaozhou Bay. (C) 2001 Academic Press.