Effects of nitrate and phosphate on growth and C2 toxin productivity of Alexandrium tamarense CI01 in culture

Growth and C2 toxin productivity of a marine dinoflagellate, Alexandrium tamarense CI01 (ATCI01) which predominantly produces C2 toxin, were studied in unialgal batch cultures to optimize the concentrations of nitrate and phosphate for a maximal toxin yield. A range of start concentrations of the two major nutrients was determined in which algal growth was proportional to the nutrient concentrations used. The highest concentrations of nitrate and phosphate in this growth-enhancing range were 264 and 20 μM, respectively. In this concentration range, the C2 toxin yield (μg/l) and cellular toxin content (Qt, fmole per cell) reached a maximum at the lowest end of phosphate (5 μM) and the highest end of nitrate (264 μM). Further increase in the supply of nitrate continued to enhance the toxin yield. Our results indicated that the growth and toxin productivity of this algal strain in batch cultures had distinctly different optimal ranges of nitrate and phosphate concentrations. For a maximum toxin yield, a judicious use of phosphate under a nitrate-replete condition is called for.     

Nitrate and phosphate supplementation to increase toxin production by the marine dinoflagellate Alexandrium tamarense

Alexandrium tamarense toxins have great value in biotechnology research as well as important in connection with shellfish poisoning. The influence of nitrate or nitrate and phosphate supplementation on cell biomass and toxin content were investigated in batch cultures. When cultures at low nitrate (88.2 microM NaNO(3)) were supplemented with 793.8 microM NaNO(3) at day 10 the cell density and cellular toxin contents were increased by 6-29% and 20-76%, respectively, compared with controls, and maximal values were 43,600 cells/ml (day 38) and 0.91 pg/cell (day 31). Supplementation with nitrate at day 14 or with nitrate and phosphate at day 10/14 to the cultures did not increase the cell density compared with the non-supplemented middle nitrate or high phosphate (108 microM NaH(2)PO(4)) cultures, respectively, but increased the cellular toxin contents by an average of 52%. The results showed that supplementation with nitrate or with nitrate and phosphate at different growth phases of the cultures increased toxin yield by an average of 46%. Supplementation with nitrate at selected times to maintain continuous low level of nitrate might contribute to the effective increase of toxin yield of A. tamarense.     

Size effects of suspended particles on gill damage in green-lipped mussel Perna viridis

The green-lipped mussels Perna viridis were exposed to <500 μm suspended solids (SS) with concentrations of 0 (control), 250, 500, 750 and 1000 mg/L for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significantly higher damages to the cilia on the frontal surface of the gill filaments than that on the abfrontal surface in both demibranchs. Percent ciliary depletion varied with SS concentrations and time. No sign of recovery of the gill filaments was observed after the mussels were transferred to clean seawater. In a second experiment, mussels were exposed to SS with size range from <63, >125–<250 and >250–<500 μm at 600 mg/L, together with a control (0 mg/L) for 14 days, followed by transferring to clean, filtered seawater for 28 days. Results of scanning microscopy showed significant ciliary damages in both the ascending and descending lamellae under the three particle size groups as compared with the control. Percent depletion of frontal cilia was most serious for the >250–<500 μm size group and least for the <63 μm size group. However, percent depletion of abfrontal cilia was most serious for the >125–<250 μm size group and least for the <63 μm size group. No recovery of ciliary damages was observed. The effects of particle size of suspended sediments on the morphological damages of gill filaments in the green-lipped mussels were discussed.     

The high content of polyunsaturated fatty acids in Nannochloropsis limnetica (Eustigmatophyceae) and its implication for food web interactions, freshwater aquaculture and biotechnology

In the eustigmatophycean Nannochloropsis limnetica the content of polyunsaturated fatty acids (PUFA) is extremely high in comparison to different planktonic green algal taxa in freshwater ecosystems. The sums of n-6 and n-3 fatty acids in N. limnetica were ten-fold higher than in the other picoplankton Choricystis minor and Pseudodictyosphaerium jurisii, and higher than in the nanoplanktonic green algae Chlorella vulgaris, Monoraphidium neglectum and Scenedesmus obtusiusculus. The content of fatty acids in N. limnetica was highly variable under different culture conditions. The highest concentrations of PUFA in N. limnetica were found in non-aerated suspension cultures, with a high content of phosphate (40 mg l⁻¹ K₂HPO₄) in the culture medium: linoleic acid 22.19 mg g⁻¹ DW, arachidonic acid 10.52 mg g⁻¹ DW, and eicosapentaenoic acid 55.56 mg g⁻¹ DW. N. limnetica represent a high-quality food resource in freshwater food chains. Furthermore, cultures of this eustigmatophycean alga have a high potential for use in biotechnology and aquaculture.     

Distribution of dissolved free amino acids, dissolved inorganic nitrogen and chlorophyll a in the surface microlayer and subsurface water of the Yellow Sea, China

Concentrations of 14 different dissolved free amino acids (DFAA), dissolved inorganic nitrogen (DIN), and chlorophyll a were determined in the surface microlayer and subsurface waters at 41 stations in the Yellow Sea, China in April 2006. The concentrations of DFAA in the subsurface water ranged from 0.13 to 1.62 μM, with an average of 0.57±0.05 μM, while those in the surface microlayer varied between 0.22 and 2.6 μM, with an average of 0.94±0.08 μM. Major constituents of DFAA present in the study area were glycine, alanine, glutamic acid, serine and histidine. One-way analysis of variance (ANOVA) showed no significant difference in average mol fractions of DFAA between microlayer and subsurface water (F=0.0440, P=0.8355). Hierarchical cluster analysis of the station similarity based on the DFAA composition in both the surface microlayer and subsurface water separated three clusters of stations at the 70% Bray–Curtis similarity level. The average concentrations of chlorophyll a and DIN were 1.18 (0.34–4.44) μg L⁻¹ and 16.57 (3.98–49.59) μM in the subsurface water, and those in the surface microlayer were 1.30 (0.24–3.97) μg L⁻¹ and 18.56 (5.77–48.93) μM, respectively. Our results showed that concentrations of chlorophyll a (r²=0.7940, n=41, p<0.0001), DIN (r²=0.6939, n=41, p<0.0001) and DFAA (r²=0.2416, n=41, P=0.0011) in the microlayer were, respectively, correlated with their subsurface water concentrations, implying that there was a strong exchange effect between the microlayer and subsurface water. The enrichment factor of DFAA in the microlayer ranged from 0.47 to 2.24 with a mean of 1.88±0.16.     

低浓度硝态氮促进微囊藻累积多聚磷酸盐

2016年以来太湖总磷浓度高位波动而总氮浓度持续下降,藻细胞内源性磷释放是湖泊水体总磷的重要来源,而多聚磷酸盐作为藻细胞内磷的储存库,其含量变化会显著影响藻细胞内源性磷的释放量.针对上述现象,开展了不同硝态氮浓度影响野外水华蓝藻及实验室纯培养的铜绿微囊藻(Microcystis aeruginosa)利用磷特别是合成多...     

Changes in the yield of chlorophyll a from dissolved available inorganic nitrogen after an enrichment event—applications for predicting eutrophication in coastal waters

An understanding of the dynamic relationship between nitrogen supply and the formation of phytoplankton biomass is important in predicting and avoiding marine eutrophication. This relationship can be expressed as the short-term yield q of chlorophyll from dissolved available inorganic nitrogen (DAIN), the sum of nitrate, nitrite and ammonium. This paper communicates the results of a continuous culture nitrate enrichment experiment undertaken to investigate the cumulative yield of chlorophyll from DAIN (q). The purposes of the study were: to acquire a better understanding of the relationship between chlorophyll formation and DAIN; to obtain values that could be used in models for predicting eutrophication. The results of a time series experiment carried out using microplankton (all organisms <200 μm in size) indicate that the parameter q does not have a single value but is affected by the ecophysiological response of phytoplankton to changing nutrient status after an enrichment event. It is also dependent on changes in the allocation of nitrogen between autotrophs and heterotrophs. The value of yield obtained at the height of the bloom can be represented by q (max) (2.35 μg chl (μmol N)⁻¹). The post-bloom, steady state value of q can be represented by q_eq (0.95 μg chl (μmol N)⁻¹). The microcosm steady state yield was not significantly different from the median value obtained from synoptic studies of Scottish west coast waters. It is proposed that q_eq is the most appropriate value for assessing the general potential for eutrophication resulting from continuous nutrient enrichment into coastal waters. It is further proposed that q (max) be used for cases of sporadic enrichment and where a short burst of unrestricted growth may be detrimental.     

Dissolved inorganic nutrients and organic substrates in the River Warnow (North-Eastern Germany) – Utilisation by bacterioplankton

After 1990, external loads of Central European rivers with inorganic nutrients (nitrogen and phosphorus) and organic material were reduced because of changed environmental laws. However, in the eutrophic lowland River Warnow, North-Eastern Germany, nitrate concentrations remained high with 35–185 μmol l⁻¹ without a significant decrease since 1992. In contrast, phosphate concentrations, varying between 0.3 and 5.2 μmol l⁻¹ during the growth season 2002, decreased significantly over the years. However, its concentrations still exceeded 1 μmol l⁻¹ regularly in the growth seasons. This load led to a substantial accumulation of organic matter additional to high terrestrial inputs. Despite the high organic load, the remineralising bacteria were mainly inactive in River Warnow. Therefore, the composition of the dissolved organic material, especially its bioavailability, were investigated during the growth season 2002 and discussed with other potential controlling factors. River Warnow carried a high load of dissolved organic carbon (14 mg l⁻¹), especially of humic substances (5.5 mg C l⁻¹). Bacterial abundance (12×10⁶ ml⁻¹) as well as production (1.7 μg C l⁻¹ h⁻¹) depended on temperature. During late spring and summer at constantly higher temperatures, bacterial production correlated positively to readily utilisable substrates and to humic compounds. Thus, the bacterial community in River Warnow may be well adapted or contain enough species using the available amino acids and carbohydrates as well as humic matter compounds. However, calculated from protozoan biomass, grazing may control bacterial biomass and perhaps community composition profoundly, what lead to the low percentages of active bacteria.     

Microcosm experiments on the effect of nutrient enrichment and a soil layer on the development of freshwater plankton

We investigated the response of phytoplankton and zooplankton to experimental alteration of nitrate and phosphate levels in outdoor enclosures. Experiments were conducted in summer and winter and in the absence and presence of a layer of soil. The tubs (12 in all) except the two plain water controls were manured initially with a mixture of fresh cowdung (50g 1⁻¹), mustard oil cake (25 g l⁻¹) and poultry wastes (25 g l⁻¹; mostly excreta), prior to enrichment. Water samples were collected from the experimental tubs twice a week to measure selected physico-chemical and biological variables. Water temperature in the summer experiments ranged from 20–30 °C and during the winter experiments from 11–15 °C. The pH values ranged from 8.0 to 9.5 and the dissolved oxygen levels from 8.2–10.0 mg l⁻¹. The levels of soluble reactive phosphorus and nitrate nitrogen ranged from undetectable levels to 1800 μg l⁻¹ and 6000 μg l⁻¹, respectively. The increase in chlorophyll-a following enrichment was rapid (3–7 days) during summer, but slower in winter (7–14 days). The predominant phytoplankton species observed in the tubs belong to the genera Sphaerocystis, Chlorella, Scenedesmus, Cosmarium, Ulothrix, Zygnema, Gonium and Pandorina. The rotifer species observed were Brachionus calyciflorus, Rotaria neptunia, Lecane bulla, L. luna, L. unguitata, Euchlanis dilatata, Asplanchna intermedia, Pseudoharringia spp., Eosphora spp., Lepadella ovalis, Epiphanes brachionus, Hexarthra mira and Cephalodella gibba. The cladocerans observed were Macrothrix spp. and Alona spp.     

Substrate influence and temporal changes on periphytic biomass accrual and metabolism in a tropical humic lagoon

We performed a field experiment in a tropical humic coastal lagoon to evaluate periphyton biomass accrual and metabolism on three different substrates (1) plastic ribbons, (2) green and (3) senescent leaves of the emergent macrophyte Typha domingensis) over 30 days. The contribution of autotrophic biomass decreased as total biomass increased over the time. Mean periphytic ash free dry weight ranged from 0.8 to 5.6 mg cm⁻², but periphyton chlorophyll a concentrations presented shorter amplitudes, which oscillated from 0.12 to 0.44 μg cm⁻² throughout the experiment. Periphyton metabolism was overall heterotrophic on all substrates, especially on senescent leaves. Our data show that substrate type influenced both biomass accrual and periphyton net productivity and respiration rates throughout periphyton development and highlighted the dominance of heterotrophic metabolism. The periphyton respiration may be subsidized by both water- and substrate-derived allochthonous energy pathways, shedding light on the role of periphytic assemblages to the carbon cycling, as a source of CO₂ to the system.     

Effects of some environmental parameters on the reproduction and development of a tropical marine harpacticoid copepod Nitocra affinis f. californica Lang

The effects of salinity, temperature, and light conditions on the reproduction and development of harpacticoid copepod, Nitocra affinis f. californica under controlled laboratory conditions were determined. Seven different salinity levels (5, 10, 15, 20, 25, 30, 35 ppt), four temperatures (20, 25, 30, 35 °C), three different light intensities (25, 56, 130 μmol m⁻² s⁻¹) and photoperiods (24 h:0 h, 1 h:23 h, 12 h:12 h LD cycle) were employed in this study. The highest (p < 0.05) overall reproduction and fastest development time were achieved by copepods reared under 30–35 ppt salinity. The optimum temperature required for the maximum reproduction was 30 °C while under 30 °C and 35 °C the copepod development time was shortest (p < 0.05) compared to other temperature levels. The overall reproduction was highest (p < 0.05) and development rate of N. affinis was shortest (p < 0.05) under lowest light intensity (25 μmol m⁻² s⁻¹). Continuous light (24 h:0 h LD) inhibited the egg production while, continuous darkness (1 h:23 h LD) and 12 h:12 h LD significantly favoured the overall reproductive activity of the female. Photoperiods 1 h:23 h and 12 h:12 h LD yielded highest total (p < 0.05) offspring female⁻¹ coupled with highest (p < 0.05) survival percentage. This study illustrated that although N. affinis can tolerate wide range of environmental conditions, prolonged exposure to subnormal environments affect its reproduction and development. This study showed that this species can be mass cultured for commercial purposes and has a potential to be used for toxicity studies due to its high reproductive performance fast development and a wide range of tolerance to environmental conditions.     

Changes in phytoplankton following nitrate and phosphate additions to large enclosures in Marion Lake,British Columbia

The addition of nitrate, phosphate and a nitrate phosphate combination to replicated wooden enclosures in Marion Lake resulted in significant increases in primary productivity and algal standing crop in each of the three types of treated enclosures. This indicated that no single nutrient is likely to simultaneously limit the growth of all phytoplankton species present at any one time in the lake epilimnion. The relevance of this to the detergent controversy is discussed. Canadian Contribution to the International Biological Programme No. CCIBP 208.     

Molecular ecological responses of dinoflagellate, Karenia mikimotoi to environmental nitrate stress

Karenia mikimotoi is one of the most important harmful algal species in the Chinese coastal waters, and which produce hemolytic toxins and ichthyotoxins, resulting in devastating economic losses. Previous studies demonstrated that the increase of nitrate concentration could promote the growth and reproduction of K. mikimotoi. However, the intrinsic mechanisms regarding the effects of nitrate on the K. mikimotoi photosynthesis, nucleic acid replication and differential protein expression remain to be elucidated. Our study demonstrated that nitrate stress inhibited growth of K. mikimotoi (p < 0.01). Algal chlorophyll fluorescence intensity varied slightly while algal cell cycle succession was significantly retarded by nitrate stress (p < 0.05). Sixteen proteins were detected only in nitrate-limited cultures which related to nitrate transport, signal transduction, amino acid metabolism, DNA repair and hemolysin manufacture. Eleven proteins were detected only in nitrate-replete sample and were related to photorespiration, reproduction and growth, assistance of protein modification, cytoskeleton stability and signal transduction. Based on analysis of differential proteomic functional annotations, we hypothesized a proteomic response mechanism of K. mikimotoi to environmental nitrate stress.     

Sulfur geochemistry of hydrothermal waters in Yellowstone National Park, Wyoming, USA. II. Formation and decomposition of thiosulfate and polythionate in Cinder Pool

Cinder Pool is an acid-sulfate-chloride boiling spring in Norris Geyser Basin, Yellowstone National Park. The pool is unique in that its surface is partially covered with mm-size, black, hollow sulfur spherules, while a layer of molten sulfur resides at the bottom of the pool (18 m depth). The sulfur speciation in the pool was determined on four different days over a period of two years. Samples were taken to evaluate changes with depth and to evaluate the importance of the sulfur spherules on sulfur redox chemistry. All analyses were conducted on site using a combination of ion chromatography and colorimetric techniques.Dissolved sulfide (H₂S), thiosulfate (S₂O₃²⁻), polythionates (S_xO₆²⁻), and sulfate were detected. The polythionate concentration was highly variable in time and space. The highest concentrations were found in surficial samples taken from among the sulfur spherules. With depth, the polythionate concentrations dropped off. The maximum observed polythionate concentration was 8 μM. Thiosulfate was rather uniformly distributed throughout the pool and concentrations ranged from 35 to 45 μM. Total dissolved sulfide concentrations varied with time, concentrations ranged from 16 to 48 μM. Sulfate was relatively constant, with concentrations ranging from 1150 to 1300 μM. The sulfur speciation of Cinder Pool is unique in that the thiosulfate and polythionate concentrations are significantly higher than for any other acid-sulfate spring yet sampled in Yellowstone National Park. Complementary laboratory experiments show that thiosulfate is the intermediate sulfoxyanion formed from sulfur hydrolysis under conditions similar to those found in Cinder Pool and that polythionates are formed via the oxidation of thiosulfate by dissolved oxygen. This last reaction is catalyzed by pyrite that occurs as a minor constituent in the sulfur spherules floating on the pool's surface. Polythionate decomposition proceeds via two pathways: (1) a reaction with H₂S, yielding thiosulfate and elemental sulfur; and (2) by disproportionation to sulfate and thiosulfate.This study demonstrates that the presence of a subaqueous molten sulfur pool and sulfur spherules in Cinder Pool is of importance in controlling the pathways of aqueous sulfur redox reactions. Some of the insights gained at Cinder Pool may be relevant to acid crater lakes where sulfur spherules are observed and variations in polythionate concentrations are used to monitor and predict volcanic activity.     

垂向归纳模型下鄱阳湖丰、枯水期初级生产力特征及与环境因子相关性分析

于2014年1月(枯水期)、7月(丰水期)对鄱阳湖湖水进行采集,测定相应的理化参数、叶绿素a浓度和光合有效辐射,结合初级生产力垂向归纳模型估算浮游植物初级生产力,分析湖区初级生产力特征及与环境因子的相关性.结果表明,鄱阳湖枯水期浮游植物初级生产力波动范围为83.50~355.43 mg C/(m~3·d),平均值为193.33 mg C/(m~3·d),初级生产力空间分布特征主要受水体类型的影响,枯水期初级生产力与氮、磷营养盐浓度呈负相关,其中与铵态氮浓度呈显著负相关,枯水期不会出现营养盐限制现象;丰水期浮游植物初级生产力波动范围为113.80~1134.06 mg C/(m~3·d),平均值为412.12 mg C/(m~3·d),初级生产力空间分布主要受河流注入的影响,丰水期浮游植物初级生产力与总磷及悬浮物浓度呈显著正相关,由于悬浮物对浮游植物生长的促进作用大于抑制作用,鄱阳湖丰水期会出现磷营养盐的限制;鄱阳湖整体平均流速约为0.28 m/s,易于浮游植物的生长,南鄱阳湖平均流速约为0.21 m/s,而北鄱阳湖平均流速约为0.35 m/s,所以南鄱阳湖比北鄱阳湖更容易发生水体富营养化并暴发水华.     

Seasonal variations in phytoplankton biomass and primary production in the Ethiopian Rift Valley lakes Ziway, Awassa and Chamo – The basis for fish production

Seasonal variations in the biomass (Chl a) and primary production (¹⁴C-method) of phytoplankton were studied during 12 months of 2005 in the three Ethiopian Rift Valley Lakes (ERVL) Ziway, Awassa and Chamo. Chl a showed an average value of 40, 20, and 30 mg m⁻³ for the three lakes, respectively. Integrated areal primary production for the total phytoplankton (g C m⁻² d⁻¹) varied 2-fold in the three lakes but on different levels, from 0.67–1.8 in L. Ziway, 1.8–4.6 in L. Awassa, and 1.0–2.6 in L. Chamo. The overall photosynthetic efficiency of utilizing photosynthetically active radiation by the phytoplankton on molar basis (mmol C mol of photons⁻¹) resulted in an average value of 1.4 for L. Ziway, 3.5 for L. Awassa and 1.6 for L. Chamo. Among the different factors regulating phytoplankton primary productivity, light penetration and nutrients were the most important in the three lakes. The seasonal variations of incident radiation (most values between 5 and 7 E m⁻² h⁻¹) and water temperature (most values between 22 and 24 °C) were small and unlikely to result in the marked differences in phytoplankton primary production. Although relative increase in nutrient concentrations occurred following the rainy periods, the major algal nutrients were either consistently low (nitrate and/or silicate) or high (phosphate and/or ammonium) and remained within a narrow range for most of the study period in all the three lakes. Consequently, phytoplankton biomass and primary production seem to be maintained more by nutrient regeneration or turnover (facilitated by high temperature) than by allochthonous nutrient input. This would be coupled with wind-induced mixing that would play an important role in determining hydrographic characteristics (water column structure) and the associated redistribution of nutrients and phytoplankton, the availability of light and subsequently the spatial (vertical) and temporal patterns of phytoplankton production in these three ERVL. Phytoplankton production (PP) is regarded as a good predictor of fish yield in lakes and seasonal measurements of PP is a prerequisite for good such estimates.     

Bio-dependent bed parameters as a proxy tool for sediment stability in mixed habitat intertidal areas

The stability of cohesive and non-cohesive sediments in a mixed intertidal habitat within the Ria Formosa tidal lagoon, Portugal, was examined during two field campaigns as part of the EU F-ECTS project. The cohesive strength meter Mk III was used to determine critical erosion shear stress (τ_c) within a variety of different intertidal habitats and substrata, including Spartina maritima fields and Zostera noltii beds. The best predictor(s) for τ_c were derived from a range of properties measured for the surface sediments (chlorophyll a, colloidal carbohydrate, water, organic content, % fraction <63 μm, and seabed elevation). Pigment biomarkers were used to identify the dominant algal groups within the surface phytobenthic assemblage.Strong, seasonally dependent relationships were found between τ_c and habitat type, chl a, colloidal carbohydrate and bed elevation. Typically, critical erosion thresholds decreased seawards, reflecting a change from biostabilisation by cyanobacteria in the upper intertidal areas, to biostabilisation by diatoms on the bare substrata of the channel edges. In the late summer/early autumn, cyanobacteria were the main sediment stabilisers, and colloidal carbohydrate was the best bio-dependent predictor of τ_c across the entire field area. In the late winter/early spring, cyanobacterial activity was lower, and sediment stabilisation by Enteromorpha clathrata was important; the best predictor of τ_c was bed elevation. The implications and use of proxies for sediment stability are discussed in terms of feedback and sedimentation processes operating across the intertidal area.     

Lake Koronia, Greece: Shift from autotrophy to heterotrophy with cultural eutrophication and progressive water-level reduction

Lake Koronia, a Ramsar site, is shallow, polymictic, hypertrophic and until recently was aerially the fourth largest lake in Greece. Although exceeding 5 m in the past, lake depth has declined progressively from 3.8 m in 1980 to < 1 m in 1997, reducing surface area and water volume by 50% and 80%, respectively. Specific conductivityincreased from 1300 μS cm⁻¹ in 1977 to >6000 μS cm⁻¹ in 1991. Increased phosphate concentrations from the late 1970's (8–45 μg L⁻¹) to the late 1990's (100–1000 μg L⁻¹) document that the previously eutrophic system with a limited littoral zone switched to hypertrophy dominated by massive cyanobacteria blooms. Oxygen saturation of the water column increased progressively from about 80% in 1983 to full saturation about 1993, after which it decreased progressively to only 20% saturation in 1997. In spite of cyanobacteria dominance, community metabolism of the lake switched from progressively increasing autotrophy to rapidly advancing heterotrophy associated with progressive water-level reduction leading to fish extirpation in the lake.