Nuclear matrix of the most primitive eukaryote Archezoa

Accumulating evidence suggests that unicellular Archezoa are the most primitive eukaryotes and their nuclei are of significance to the study of evolution of the eukaryotic nucleus. Nuclear matrix is an ubiquitous important structure of eukaryotic nucleus; its evolution is certainly one of the most important parts of the evolution of nucleus. To study the evolution of nuclear matrix, nuclear matrices of Archezoa are investigated. Giardia lamblia cells are extracted sequentially. Both embedment-free section EM and whole mount cell EM of the extracted cells show that, like higher eukaryotes, this species has a residual nuclear matrix in its nucleus and rich intermediate filaments in its cytoplasm, and the two networks connect with each other to form a united network. But its nuclear matrix does not have nucleolar matrix and its lamina is not as typical as that of higher eukaryotes; Western blotting shows that lamina of Giardia and two other Archezoa Entamoeba invadens and Trichomonas vaginali all contain only one polypeptide each which reacts with a mammalia anti-lamin polyclonal serum and is similar to lamin B (67 ku) of mammlia in molecular weight. According to the results and references, it is suggested that nuclear matrix is an early acquisition of the eukaryotic nucleus, and it and the "eukaryotic chromatin" as a whole must have originated very early in the process of evolution of eukaryotic cell, and their origin should be an important prerequisite of the origin of eukaryotic nucleus; in the iamin (gene) family, B-type lamins (gene) should be the ancestral type and that A-type lamins (gene) might derive therefrom.

The nuclear matrix of Euglena gracilis (Euglenophyta): A stage of nuclear matrix evolution?

Euglena gracilis cell was extracted sequentially with CSK-Triton buffer, RSB-Magik solution and DNase-As solution. DGD embedment-free electron microscopy showed that in the extracted nucleus there was a residual non-chromatin fibrous network. That it could not be removed by hot trichloroacetic acid further supported the idea that it was a non-histone, non-chromatin fibrous protein network, and should be the internal network of the nuclear matrix. After the sequential extraction, the nuclear membrane was removed, leaving behind a layer of lamina; the chromatin was digested and eluted from the dense chromosomes and residual chromosomal structures that should be chromosomal scaffold were revealed. Western blot analysis with antiserum against rat lamins showed that nuclear lamina of the cell possessed two positive polypeptides, a major one and a minor one, which had molecular masses similar to lamin B and lamin A, respectively. Comparing these data with those of the most primitive eukaryote Archezoa and of higher eukaryotes, it was suggested that the lower unicellular eukaryote E. gracillis already had the nuclear matrix structure, and its nuclear matrix (especially the lamina) might represent a stage of evolutionary history of the nuclear matrix. (C) 2000 Editions scientifiques et medicales Elsevier SAS.

Mitigation of lake eutrophication: Loosen nitrogen control and focus on phosphorus abatement

w Traditionally, nitrogen control is generally considered an important component of reducing lake eutrophication and cyanobacteria blooms. However, this viewpoint is refuted recently by researchers in China and North America. In the present paper, the traditional viewpoint of nitrogen control is pointed out to lack a scientific basis: the N/P hypothesis is just a subjective assumption; bottle bioassay experiments fail to simulate the natural process of nitrogen fixation. Our multi-year comparative research in more than 40 Yangtze lakes indicates that phosphorus is the key factor determining phytoplankton growth regardless of nitrogen concentrations and that total phytoplankton biomass is determined by total phosphorus and not by total nitrogen concentrations. These results imply that, in the field, nitrogen control will not decrease phytoplankton biomass. This finding is supported by a long-term whole-lake experiment from North America. These outcomes can be generalized in terms that a reduction in nitrogen loading may not decrease the biomass of total phytoplankton as it can stimulate blooms of nitrogen-fixing cyanobacteria. To mitigate eutrophication, it is not nitrogen but phosphorus that should be reduced, unless nitrogen concentrations are too high to induce direct toxic impacts on human beings or other organisms. Finally, details are provided on how to reduce controls on nitrogen and how to mitigate eutrophication. (C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Assessment effects of cage culture on nitrogen and phosphorus dynamics in relation to fallowing in a shallow lake in China

Nitrogen and phosphorus dynamics in relation to fallowing in a fish cage farm was investigated in a shallow lake in China. Four sampling sites were set: beneath the cages, at the cage sides, and 50 and 100 m east of the cage farm. Total nitrogen (TN) and total phosphorus (TP) in lake water and sediment were analyzed during a 2-year rearing cycle. The cage culture had a fish yield of 16.3-39.2 tonnes in the study period. Based on the mass balance equation, 1533-3084 kg TN and 339-697 kg TP were contributed to the lake environment. Nitrogen and phosphorous concentrations showed greater increase in the first culture period than in the second rearing cycle. No obvious changes were found at the sampling sites 50 and 100 m east of the cages during the study periods. Main impacts were found close to the cages (beneath the cages and at the cage side); the sampling points at the cage side showed relatively high TN and TP sedimentation. After 3 months of fallowing, water TN and TP decreased significantly but the sediment TN and TP contents remained high. Therefore, recovery seems to happen during fallowing but attention should be paid to whether the culture continues to operate in the future.

Carbon and nitrogen metabolism of an eutrophication tolerative macrophyte, Potamogeton crispus, under NH4+ stress and low light availability

Submersed macrophytes in eutrophic lakes often experience high NH4+ concentration and low light availability in the water column. This study found that an NH4+-N concentration of 1 mgL(-1) in the water column apparently caused physiological stress on the macrophyte Potamogeton crispus; L The plants accumulated free amino acids (FAA) and lost soluble carbohydrates (SC) under NH4+ stress. These stressful effects of NH4+ were exacerbated under low light availability. Shading significantly increased NH4+ and FAA contents and dramatically decreased SC and starch contents in the plant shoots. At an NH4+-N concentration of 1 mg L-1 in the water column, neither growth inhibition nor NH4+ accumulation was observed in the plant tissues of P. crispus under normal light availability. The results showed that 1 mg L-1 NH4+-N in the water column was not toxic to P. crispus in a short term. To avoid NH4+ toxicity. active NH4+ transportation out of the cell may cost energy and thus result in a decline of carbohydrate. When NH4+ inescapably accumulates in the plant cell, i.e. under NH4+ Stress and shading, NH4+ is scavenged by FAA synthesis. (c) 2009 Published by Elsevier B.V.

Nitrogen dynamics in large shallow eutrophic Lake Chaohu, China

Temporal and spatial dynamics of nitrogen in lake and interstitial water were studied monthly in a large shallow, eutrophic lake in subtropical China from October 2002 to September 2003. The distribution of nitrogen was consistent with the idea that high nitrogen concentrations in the western part of the lake resulted from high levels of the nutrients from the surrounding cities through sewage-drainage systems. Nitrate was the predominant form of nitrogen in the overlying water, while ammonium was predominant in the interstitial water, indicating that strong oxidative nutrient regeneration occurred near the sediment-water interface. Nitrate could be an important dissolved inorganic matter source for phytoplankton, which in turn influenced the seasonal variations of nitrate concentrations in lake water. Significant positive correlation between ammonium fluxes and water temperature was observed and could probably be attributed to the intensified ammonification and nitrate reduction with increased temperature. Positive correlation between ammonium fluxes and algae biomass and Chl a concentrations may indicate that phytoplankton was an important factor driving ammonium fluxes in our study lake, and vice versa that higher fluxes of ammonium supported a higher biomass of the phytoplankton.

Effects of ammonium on growth, nitrogen and carbohydrate metabolism of Potamogeton maackianus A. Benn

Growth, nitrogen and carbohydrate metabolism in relation to eutrophication were studied for a submerged plant Potamogeton maackianus, a species common in East Asian shallow lakes. The plants were grown in six NH4+-N concentrations (0.05, 0.50, 1.00, 3.50, 5.00 and 10.00 mg/L) for six days. NH4+-N levels in excess of 0.50 mg/L inhibited the plant growth. The relationships between external NH4+-N availability and total nitrogen (TN), protein-N, free amino acid-N (FAA-N) and NH4+-N in plant tissues, respectively, conformed to a logarithmic model suggesting that a feedback inhibition mechanism may exist for ammonium uptake. The response of starch to NH4+-N was fitted with a negative, logarithmic curve. Detailed analysis revealed that the influx NH4+-N had been efficiently incorporated into organic-N and eventually stored as protein at the expense of starch accumulation. These data suggest that this species may be able to tolerate high levels of ammonium when dissolved oxygen is sufficient.

Differentiate performance of eight filter media in vertical flow constructed wetland: Removal of organic matter, nitrogen and phosphorus

A comparative study was conducted to reveal the differentiate effects of eight different filter media including gravel, zeolites, anthracite, shale, vermiculite, ceramic filter media, blast furnace steel slag and round ceramsite. The study mainly related to the eight different filter media's removal performances of organic matter, nitrogen and phosphorus in the vertical flow constructed wetland simulated system, which treating wastewater at hydraulic loading rate of 1000-2500 mm/d. The results indicated that the removal effects were closely related to the physical and chemical properties of medium materials. Anthracite-filled system had the highest removal rate for the total organic carbon (TOC), up to 70%, and the removal rates of other systems ranged from 20% to 30%. As for the five-day biochemical oxygen demand (BOD5), anthracite-filled and steel slag-filled systems had the highest removal rates, also up to 70%, as well as other systems all exceeded 50%. At the same time, for the total nitrogen (TN) and NH4(+)-N, the zeolites-filled and ceramic-filled systems had the best performances with the removal rates of more than 70%, the other way round, the removal rates of other systems were only about 20%. The distinguishable effects were also observed in removal performances of total phosphorus (TP) and total dissoluble phosphorus (TDP). The removal rates of TP and TDP in steel slag-filled systems were more than 90%, a much higher value, followed by that of the anthracite-filled system, more than 60%, but those of other systems being the less. Our study provided a potential mechanism to optimize the filter media design for the vertical flow constructed wetlands.

Icefish (Salangidae) as an indicator of anthropogenic pollution in freshwater systems using nitrogen isotope analysis

We investigated differences in delta N-15 of seston and icefishes from seven freshwater ecosystems with different trophic states in China. An increase of seston delta N-15 values was accompanied by an increase of total nitrogen and phosphorus concentrations. Significantly positive correlations were observed between delta N-15 of icefishes and delta N-15 of seston, total nitrogen and phosphorus concentrations. This study demonstrated that icefishes could be preferred indicators of anthropogenic contamination in test systems because they integrated waste inputs over long time periods and reflected the movement of waste through the pelagic food chain.

Meat and bone meal replacement in diets for juvenile gibel carp (Carassius auratus gibelio): effects on growth performance, phosphorus and nitrogen loading

A 11-week growth trial was conducted in a flow-through system with juvenile gibel carp Carassius auratus gibelio to evaluate the effects of gradual replacement of fish meal (FM) by meat and bone meal (MBM) on growth performance, phosphorus (P) and nitrogen (N) loading. Six isonitrogenous (crude protein: 410 g kg(-1)) and isoenergetic (gross energy: 18 kJ g(-1)) diets were formulated. FM was used as the control protein. In the other five diets, 20, 40, 60, 80 and 100% FM protein was substituted with MBM20, MBM40, MBM60, MBM80, MBM100, respectively. Total P content in the diets ranged from 16.0 to 28.3 g kg(-1) and the available P was 5.0-6.6 g kg(-1). The results showed that the best growth was achieved with fish fed on the control diet and MBM20. Final body weight, weight gain, feed efficiency, protein retention efficiency and energy retention efficiency decreased with increased dietary MBM. No significant differences were found in the feeding rate and hepatosomatic index between the groups. Apparent digestibility coefficient (ADC) of dry matter, protein and P decreased with increase in dietary MBM, while there were no significant differences in the ADC of energy. P and N retention decreased linearly while P and N loading increased linearly with the increased dietary MBM levels. No significant differences were observed in the activity of alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase, as well as pyruvate kinase in liver or in serum. Total superoxide dismutase activity in MBM20 was significantly higher than that of MBM100.

Distribution of nitrogen and phosphorus in sediment cores and overlying water of Donghu Lake

In this study, the seasonal, vertical distribution of various phosphorus and nitrogen forms in the sediment and overlying water of Donghu Lake were investigated. The concentration of total nitrogen (TN) in overlying water was high in spring and autumn, but that of NO3--N reached its peak in autumn. From summer to autumn and from winter to spring, the concentration of phosphorus in overlying water decreased, while it increased from autumn to winter. Vertical characteristic forms of phosphorus in sediment cores are total phosphorus (TP), labile phosphorus (LP), Fe-P and Al-P, obviously enriched in the surface layer (0-10 cm), but their concentrations are observably reduced along with the depth of sediment. The research is of important theoretical and practical value to understand the status and to control the developmental trend of eutrophication in Donghu Lake.

Changes in the patterns of inorganic nitrogen and TN/TP ratio and the associated mechanism of biological regulation in the shallow lakes of the middle and lower reaches of the Yangtze River

The changes of NH3-N, NO3-N, NO2-N and TN/TP were studied during growth and non-growth season in 33 subtropical shallow lakes in the middle and lower reaches of the Yangtze River. There were significant positive correlations among all nutrient concentrations, and the correlations were better in growth season than in non-growth season. When TP > 0.1 mgL(-1), NH3-N increased sharply in non-growth season with increasing TP, and NO3-N increased in growth season but decreased in non-growth season with TP. These might be attributed to lower dissolved oxygen and low temperature in non-growth season of the hypereutrophic lakes, since nitrification is more sensitive to dissolved oxygen and temperature than anti nitrification. When 0.1 mgL(-1)> TP > 0.035 mgL(-1), TN and all kinds of inorganic nitrogen were lower in growth season than in non-growth season, and phytoplankton might be the vital regulating factor. When TP < 0.035 mgL(-1), inorganic nitrogen concentrations were relatively low and NH3-N, NO2-N had significant correlations with phytoplankton, indicating that NH3-N and NO2-N might be limiting factors to phytoplankton. In addition, TN/TP went down with decline in TIP concentration, and TN and inorganic nitrogen concentrations were obviously lower in growth season than in non-growth season, suggesting that decreasing nitrogen (especially NH3-N and NO3-N) was an important reason for the decreasing TN/TP in growth season. The ranges of TN/TP were closely related to trophic level in both growth and non-growth seasons, and it is apparent that in the eutrophic and hypertrophic state the TN/TP ratio was obviously lower in growth season than in non-growth season. The changes of the TN/TP ratio were closely correlated with trophic levels, and both declines of TN in the water column and TP release from the sediment were important factors for the decline of the TN/TP ratio in growth season.

Sedimentation rates, nitrogen and phosphorus retentions in the largest urban Lake Donghu, China

Sediment core samples were collected in the largest urban Lake Donghu (Stations I and II) in China, and the activities of Pb-210, Ra-226 and Cs-137 were measured by gamma-ray spectrometry. The sedimentation rates, calculated by 210Pb constant rate of supply (CRS) model, ranged from 0.11 to 0.65 (average 0.39) cm(.)y(-1) at Station I, and from 0.21 to 0.78 (average 0.46) cm(.)y(-1) at Station II. Sedimentation rate calculated by Cs-137 as a time marker was 0.55 cm(.)y(-1) at Station II. Based on the average sedimentation rate, we obtained 769 and 147 t(.)y(-1) for nitrogen and phosphorus retentions in Lake Donghu sediments, respectively.

Effects of animal-plant protein ratio in extruded and expanded diets on nitrogen and energy budgets of juvenile Chinese soft-shelled turtle (Pelodiscus sinensis Wiegmann)

In this study, we investigated the effects of animal-plant protein ratio in extruded and expanded diets on nutrient digestibility, nitrogen and energy budgets of juvenile soft-shelled turtle (Pelodiscus sinensis). Four extruded and expanded feeds (diets 1-4) were formulated with different animal-plant protein ratios (diet 1, 1.50:1; diet 2, 2.95:1; diet 3, 4.92:1; diet 4, 7.29:1). The apparent digestibility coefficients (ADCs) of dry matter and crude lipid for diet 1 were significantly lower than those for diets 2-4. There was no significant difference in crude protein digestibility among diets 1-4. The ADC of carbohydrate was significantly increased with the increase in animal-plant protein. Although nitrogen intake rate, faecal nitrogen loss rate and excretory nitrogen loss rate of turtles fed diet 1 were significantly higher than those fed diets 2-4, nitrogen retention rate, net protein utilization and biological value of protein in these turtles were significantly lower than those fed diets 2-4. In addition, energy intake rate, excretory energy loss rate and heat production rate of turtles fed diet 1 were also significantly higher than those fed diets 2-4. Faecal energy loss was significantly reduced with the increase in the animal-plant protein ratio. The ADC of energy and assimilation efficiency of energy significantly increased with a higher animal-plant protein ratio. The growth efficiency of energy in the group fed diet 1 was significantly lower than those in the groups fed diets 2-4. Together, our results suggest that the optimum animal-plant protein ratio in extruded and expanded diets is around 3:1.

Determination of 266 pesticide residues in apple juice by matrix solid-phase dispersion and gas chromatography-mass selective detection

A macro matrix solid-phase dispersion (MSPD) method was developed to extract 266 pesticides from apple juice samples prior to gas chromatography-mass selective detection (GC-MSD) determination. A 10 g samples was mixed with 20 g diatomaceous earth. The mixture was transferred into a glass column. Pesticide residues were leached with a 160 mL hexane-dichloromethane (1:1) at 5 mL/min. Two hundred and sixty-six pesticides were divided into three groups and detected by GC-MSD under selective ion monitoring. The proposed method takes advantage of both liquid-liquid extraction and conventional MSPD methods. Application was illustrated by the analysis of 236 apple juice samples produced in Shaanxi province China mainland this year. (C) 2004 Elsevier B.V. All rights reserved.