The effect of different dietary iron levels on growth and hepatic iron concentration in juvenile gibel carp (Carassius auratus gibelio)

P>An 83-day growth trial was conducted using a flow-through system to examine the effects of different dietary iron levels on growth and hepatic iron concentration in juvenile gibel carp (Carassius auratus gibelio). Six purified diets supplemented with different levels of iron (0, 10, 30, 60, 100 and 200 mg kg(-1)) (as ferrous sulfate) were fed to triplicate groups of fish (initial weight 2.12 +/- 0.00 g per fish). The results showed that the addition of iron to the basal diet did not significantly affect the specific growth rate (SGR), feed efficiency (FE), survival, red blood cell amount (RBC), hemoglobin content (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) or mean corpuscular hemoglobin concentration (MCHC). Hepatic iron concentration and hematocrit (Hct) were significantly influenced by dietary iron level (P < 0.05). On the basis of the iron concentration for the maintenance of optimum hepatic iron concentration and Hct, it was concluded that the dietary iron concentration of juvenile gibel carp should be not less than 202 mg Fe kg(-1) diet.

Genome-wide identification and characterization of cytochrome P450 monooxygenase genes in the ciliate Tetrahymena thermophila

Background: Cytochrome P450 monooxygenases play key roles in the metabolism of a wide variety of substrates and they are closely associated with endocellular physiological processes or detoxification metabolism under environmental exposure. To date, however, none has been systematically characterized in the phylum Ciliophora. T. thermophila possess many advantages as a eukaryotic model organism and it exhibits rapid and sensitive responses to xenobiotics, making it an ideal model system to study the evolutionary and functional diversity of the P450 monooxygenase gene family. Results: A total of 44 putative functional cytochrome P450 genes were identified and could be classified into 13 families and 21 sub-families according to standard nomenclature. The characteristics of both the conserved intron-exon organization and scaffold localization of tandem repeats within each P450 family clade suggested that the enlargement of T. thermophila P450 families probably resulted from recent separate small duplication events. Gene expression patterns of all T. thermophila P450s during three important cell physiological stages (vegetative growth, starvation and conjugation) were analyzed based on EST and microarray data, and three main categories of expression patterns were postulated. Evolutionary analysis including codon usage preference, sit-especific selection and gene-expression evolution patterns were investigated and the results indicated remarkable divergences among the T. thermophila P450 genes. Conclusion: The characterization, expression and evolutionary analysis of T. thermophila P450 monooxygenase genes in the current study provides useful information for understanding the characteristics and diversities of the P450 genes in the Ciliophora, and provides the baseline for functional analyses of individual P450 isoforms in this model ciliate species.

Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry

Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced. and these sequence data were deposited in the GenBank. Eight oligonucleotide probes (DNA probes) were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order: PM18S02 > PM28S02 > PM28S01 > PM18S01, and that of the probes specific to T. pulchella was TP18S02 > TP28S01 > TP28S02 > TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02; TP18S02 and TP28S01, and the protocol, were also useful for the detection of algae in natural samples.

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.

Hydrolysis and coagulation behavior of polyferric sulfate and ferric sulfate

The hydrolysis behaviors of polyferric sulfate (PFS) and ferric sulfate (FS) under conditions similar to raw wastewater were investigated and the coagulation of biologically pretreated molasses wastewater using PFS and FS was evaluated by studying coagulation efficiency, zeta potential and microscopic surface morphology of flocs. Experimental results show that the hydrolysis behavior of PFS is different from that of FS on the basis of ferron assay. In the case of FS, fast-reacting Fe(III) polymers were the dominant polynuclear species while large fraction of slow-reacting iron polymers is present in PFS. Despite slightly fewer dosages of PFS required as compared to FS, there is no marked difference in the coagulation of molasses effluent between PFS and FS, especially at the optimum dosages. Both coagulants destabilize organic compounds predominantly through charge neutralization-precipitation mechanism. Hydrolysis rate of PFS in synthetic solution is appreciably different from that in raw wastewater. This may due to the effect of sulfate anion introduced as counter-ion as well as depolymerization of larger polymeric Fe(III) species by the organic ligands present in molasses effluent.

Mouse Toxicity of Anabaena flos-aquae from Lake Dianchi, China

Some species of the genera Anabaena can produce various kinds of cyanotoxins, which may pose risks to environment and human health. Anabaena has frequently been observed in eutrophic freshwater of China in recent years, but its toxicity has been reported only in a few studies. In the present study, the toxicity of an Anabaena flos-aquae strain isolated from Lake Dianchi was investigated. Acute toxicity testing was performed by mouse bioassay using crude extracts from the lyophilized cultures. The mice exposed to crude extracts showed visible symptoms of toxicity and died within 10-24 h of the injection. Serum biochemical parameters were evaluated by the use of commercial diagnostic kits. Significant alterations were found in the serum biochemical parameters: alkaline phosphatase (AKP), gamma-glutamyl transpepticlase (gamma-GT), aspartate amino transferase (AST), alanine amino transferase (ALT), AST/ALT ratio, total protein content, albumin content, albumin/globulin (A/G) ratio, blood urea nitrogen (BUN), serum creatinine (Ssr), and total antioxidative capacity (T-AOC). Histopathological observations were carried out with hematoxylin and eosin (HE) stain under light microscope. Severe lesions were seen in the livers, kidneys, and lungs of the mice injected with crude extracts. The alterations of biochemical parameters were in a dose-dependent manner, and the severities of histological lesions were in the same manner. Based on biochemical and histological studies, this research firstly shows the presence of toxin-producing Anabaena species in Lake Dianchi and the toxic effects of its crude extracts on mammals. (C) 2008 Wiley Periodicals, Inc. Environ Toxicol 24: 10-18, 2009.

Multiple cohorts of juvenile jack mackerel Trachurus japonicus in waters along the Tsushima Warm Current

The jack mackerel, Trachurus japonicus, has a prolonged spawning season and widely spread spawning grounds. The population in the coastal waters of Japan seems to be composed of several cohorts spawned seasonally from different waters. To understand its population structure along the Tsushima Warm Current, we analysed hatchdates and growth histories of fish from Kunda Bay, the southern, central and northern East China Sea (ECS), the southern Sea of Japan, and Maizuru Bay. Seven cohorts were detected from fish collected between June 2005 and June 2006 in Kunda Bay. Comparing hatchdate distributions and growth trajectories of the seven cohorts with those of the other five regional samples, we did not find that cohorts collected in Kunda Bay originated in the southern ECS. Therefore, these coastal waters of Japan appear to be significant spawning grounds for juvenile jack mackerel. (C) 2008 Elsevier B.V. All rights reserved.

The role of cell-surface-bound phosphatases in species competition within natural phytoplankton assemblage: an in situ experiment

Despite it is widely acknowledged that the ability to hydrolyze dissolved organic matter using extracellular phosphatases is diverse in fresh water phytoplankton, the competition within single species related to presence and quantity of cell-surface-bound phosphatases has not been examined in natural conditions yet. Here, we studied phytoplankton species competition in a freshwater reservoir during an in situ experiment. A natural plankton community, with the exclusion of large zooplankton, was enclosed in permeable dialysis bags inside two large containers of different bioavailable phosphate concentrations. Phytoplankton species biomass and the abundance of bacteria were determined in purpose to compare the development of enclosed microbial communities. Total and cell-surface-bound phosphatase activities in the phytoplankton were investigated using the Fluorescently Labelled Enzyme Activity (FLEA) technique that allows for direct microscopic detection of phosphatase-positive cells and, with image cytometry, enables quantification of phosphatase hydrolytic capacity. Production of extracellular phosphatases was not completely inhibited or stopped in the phosphate-enriched environment, phytoplankton cells only showed the activity less often. Under the phosphate-nonenriched conditions, the production of phosphatases was enhanced, but active species did not proliferate amongst phytoplankton assemblage. Further, specific growth rates of the phosphatase-positive species in the non-enriched environment were lower than the same phosphatase-positive species in phosphate-enriched environment. Interestingly, the phosphatase-positive cells of Ankyra ancora increased their size in both treatments equally, although the population in phosphate-enriched environment grew much faster and the cell-specific phosphatase activity was lower. We hypothesize that brand new daughter cells had sufficient phosphorus reserves and therefore did not employ extracellular phosphatases until they matured and needed extra bioavailable phosphorus to support their metabolism before cell division. Based on presented in situ experiment, we propose that the ability to hydrolyze organic polymers and particles with cell-surface-hound phosphatases is advantageous for longer persistence of given population in a phosphate-scarce environment; although phosphatase-positive species cannot dominate the reservoir phytoplankton solely because of specific phosphorus-scavenging strategy.

Effects of gibberellin A(3) on growth and microcystin production in Microcystis aeruginosa (cyanophyta)

Environmental. factors that affect the growth and microcystin production of microcystis have received worldwide attention because of the hazards microcystin poses to environmental safety and public health. Nevertheless, the effects of organic anthropogenic pollution on microcystis are rarely discussed. Gibberellin A(3) (GA(3)) is a vegetable hormone widely used in agriculture and horticulture that can contaminate water as an anthropogenic pollutant. Because of its common occurrence, we studied the effects of GA3 on growth and microcystin production of Microcystis aeruginosa (M. aeruginosa) PCC7806 with different concentrations (0.001-25mg/L) in batch culture. The control was obtained without gibberellin under the same culture conditions. Growth, estimated by dry weight and cell number, increased after the GA3 treatment. GA3 increased the amounts of chlorophyll a, phycocyanin and cellular-soluble protein in the cells of M. aeruginosa PCC7806, but decreased the accumulation of water-soluble carbohydrates. In addition, GA3 was observed to affect nitrogen absorption of the test algae, but to have no effect on the absorption of phosphorus. The amount of microcystin measured by enzyme-Linked immunosorbent assay (ELISA) increased in GA3 treatment groups, but the stimulatory effects were different in different culture phases. It is suggested that GA3 increases M. aeruginosa growth by stimulating its absorbance of nitrogen and increasing its ability to use carbohydrates, accordingly increasing cellular pigments and thus finally inducing accumulation of protein and microcystin. (C) 2007 Elsevier GmbH. All rights reserved.

The accumulation of alkylphenols in submersed plants in spring in urban lake, China

The concentrations of alkylphenols (APs) were investigated in water, sediments and submersed macrophytes from the Moon Lake, Wuhan city, China. The water samples contained APs, ranging up to 26.4 mu g l(-1) for nonylphenol (NP) and 0.68 mu g l(-1) for octylphenol (OP). APs were found in the sediment samples with concentrations ranging from 4.08 to 14.8 for NP and from 0.22 to 1.25 mu g l(-1) dry weight for OP. The samples from the site near former sewage inlet showed the highest concentrations of APs in both water and sediments. The results of distribution pattern and dynamics of NP and OP in submersed macrophytes of the Moon Lake showed that the two pollutants were all found in Myriophyllum verticillatum, Elodea nuttallii, Ceratophyllum oryzetorum, and Potamageton crispus collected from the Moon Lake. For NP, M. verticillatum had the highest capacity of accumulation, followed by E. nuttallii, C. oryzetorum and P. crispus. However the distribution pattern of OP differed from that of NP, and the highest amount of accumulation was observed in E. nuttallii, followed by M. verticillatum, P. crispus, and C. oryzetorum. The temporal pattern of APs was also observed in submersed macrophytes from March to May, and the highest accumulation period was in May. (c) 2008 Elsevier Ltd. All rights reserved.

Cloning, characterization, and gene expression analysis of a novel cadmium metallothionein gene in Tetrahymena pigmentosa

A novel cadmium-inducible metallothionein (MT) gene (Tpig-MT1) was cloned and sequenced from the ciliate Tetrahymena pigmentosa. The number of deduced amino acids is 118. The polypeptide possesses CCC and CC clusters characteristic of typical Tetrahymena Cd-inducible MTs. The structure of Tpig-MT1 is different from the reported Cd-MT in T. pyriformis, T. thermophila and T. pigmentosa. Tpig-MT1 contains two intragenic tandem repeats with 72.9% identity described as Tpig-MT1 (repeat A1) and Tpig-MT1 (repeat A2). The transcriptional response of Tpig-MT1 gene to different heavy metals (Cd, Cu, Zn, Hg, Pb) and oxidative stress (H2O2) was measured using real-time quantitative PCR. The results showed that the gene was quickly induced (1 h) by the five heavy metals and the order of expression level was Hg>Pb>Cd>Cu>Zn. The induction effect of H2O2 was 5-fold after about 15 min, but soon decreased to a non-significant level (30 min). The genetic diversity of Tetrahymena MT genes is discussed in relation to the unique structure of the Tpig-MT1 gene and other reported Cd-MT isoforms. (C) 2008 Elsevier B.V. All rights reserved.

Cytotoxicity evaluation of three pairs of hexabromocyclododecane (HBCD) enantiomers on Hep G2 cell

Hexabromocyclododecanes (HBCDs) are additive brominated flame retardants mainly used in plastics and textiles. At the present time, these compounds are found in almost all environmental and human samples. In order to evaluate the environmental safety and health risk of HBCDs, the enantiomerically pure alpha-, beta-, and gamma-HBCD were prepared using high performance liquid chromatography (HPLC) on a PM-P-CD column and the cytotoxicities of their enantiomers were evaluated in Hep G2 cells. Results from the 3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), resazurin reduction and lactate dehydrogenase (LDH) release assays showed a good agreement that the order of cytotoxicity was gamma-HBCD >= beta-HBCD > alpha-HBCD, and that significantly lower cell viability and higher LDH release were observed in all (+)-enantiomers ((+) alpha-, (+) beta- and (+) gamma-HBCD) than the corresponding (-)-forms ((-) alpha-, (-) beta- and (-) gamma-HBCD). Additionally, the formation of reactive oxygen species (ROS) induced by these HBCD enantiomers were detected. The positive correlation between the LDH release and ROS formation demonstrated that the toxic mechanism might be mediated by oxidative damage. These results suggest that environmental and human health risks of HBCDs must be evaluated at the level of individual enantiomers. (C) 2008 Published by Elsevier Ltd.

Effects of solar UV radiation on diurnal photosynthetic performance and growth of Gracilaria lemaneiformis (Rhodophyta)

Previous studies on diurnal photosynthesis of macroalgal species have shown that at similar levels of photosynthetically active radiation (PAR, 400-700nm) the photosynthetic rate is lower in the afternoon than in the morning. However, the impacts of solar ultraviolet radiation (UVR, 280-400nm) have been little considered. We investigated the diurnal photosynthetic behaviour of the economically significant red alga Gracilaria lemaneiformis in the absence or presence of UV-A+B or UV-B with a flow-through system. While UV-A and UV-B, respectively, inhibited noontime Pmax by 22% and 14% on the sunny days, UV-A during sunrise (PAR below about 50Wm-2) increased the net photosynthesis by about 8% when compared with PAR alone. UV-A + PAR also resulted in higher apparent photosynthetic efficiency in the morning than in the afternoon period than PAR alone. Nevertheless, integrated daytime photosynthetic production under solar PAR alone was higher than with either PAR + UV-A+B or PAR + UV-A. Relative growth rate in the long term (9 days) matched the integrated photosynthetic production in that UV-A led to 9-15% and UV-B to 19-22% reduction, respectively. UV-absorbing compounds were found to be higher in the thalli exposed to PAR+UV-A+B than under PAR alone, reflecting a protective response to UVR.

Effects of dietary manganese on growth and tissue manganese concentrations of juvenile gibel carp, Carassius auratus gibelio

A 68-day growth trial was conducted in a flow-through system to determine the effect of dietary manganese levels on growth and tissue manganese concentration of juvenile gibel carp (Carassius auratus gibelio). Seven purified diets containing 7.21, 8.46, 9.50, 10.50, 13.03, 19.72 and 22.17 mg manganese (as manganic sulfate) per kilogram diet were fed to triplicate groups of fish (initial weight 3.21 +/- 0.01 g). The results showed that dietary manganese levels did not significantly affect feed intake of the fish. Specific growth rate, feed efficiency, total hepatic superoxide dismutase activity, carcass and skeletal manganese concentration increased significantly with increased dietary manganese(P < 0.05) while condition factor decreased significantly(P < 0.05). It was concluded that dietary requirement of manganese was 13.77 mg Mn per kilogram diet. Carcass and skeletal manganese concentration could also be used to evaluate the manganese requirement. Total hepatic superoxide dismulase activity was not a sensitive indicator for dietary requirement.

First-principles LDA plus U and GGA plus U study of neptunium dioxide

We have performed a systematic first-principles investigation to calculate the electronic structures, mechanical properties, and phonon-dispersion curves of NpO2. The local-density approximation+U and the generalized gradient approximation+U formalisms have been used to account for the strong on-site Coulomb repulsion among the localized Np 5f electrons. By choosing the Hubbard U parameter around 4 eV, the orbital occupancy characters of Np 5f and O 2p are in good agreement with recent experiments [A. Seibert, T. Gouder, and F. Huber, J. Nucl. Mater. 389, 470 (2009)]. Comparing to our previous study of ThO2, we note that stronger covalency exists in NpO2 due to the more localization behavior of 5f electrons of Np in line with the localization-delocalization trend exhibited by the actinides series.