An enzyme-linked immunosorbent assay for rare minnow (Gobiocypris rarus) vitellogenin and comparison of vitellogenin responses in rare minnow and zebrafish (Danio rerio)

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to determine vitellogenin (Vtg) in rare minnow (Gobiocypris rarus) based on the separation and purification of rare minnow Vtg (r-Vtg) as well as the production of polyclonal antibody against r-Vtg in rabbits. Three different ELISAs for measuring r-Vtg were then compared: (1) indirect ELISA with the antibody against carp (Cyprinus carpio) Vtg (c-Vtg) (IC-ELISA); (2) competitive ELISA with the antibody against c-Vtg, and using r-Vtg for coating the plates and preparing standard curve (CC-ELISA); (3) competitive ELISA with the antibody against r-Vtg, and using r-Vtg for coating the plates and preparing standard curve (CR-ELISA). The result showed that the homologous CR-ELISA was the most sensitive among the three assays for quantifying r-Vtg. The sensitivities to 17 alpha-ethinylestradiol (EE2) Of rare minnow and zebrafish (Danio rerio) were compared upon the establishment of homologous competitive ELISA. The lowest observed effect concentrations (LOECs) to induce Vtg were found to be 0.8 ng EE2 l(-1) for rare minnow and 4 ng EE2 l(-1) for zebrafish respectively. Afterwards, CR-ELISA was applied to measure Vtg concentration in whole body homogenate (WBH) of juvenile rare minnow fed by three diets (tubifex from wastewater treatment plant, Artemia nauplii and commercial pellet food), and the agreements between bioassay and GC-MS analysis demonstrated that rare minnow was a sensitive fish model for assessing estrogenic effects of endocrine disrupting compounds in aquatic environment. (c) 2006 Elsevier B.V. All rights reserved.

Differential expression of three Paralichthys olivaceus Hsp40 genes in responses to virus infection and heat shock

Heat shock proteins (Hsps) are a family of highly conserved cellular proteins present in all organisms, mediating a range of essential housekeeping and cytoprotective functions as well-known molecular chaperons and recently as regulators of the immune response. By subtractive suppression hybridization, three Hsp40 homologues have been identified in the flounder (Paralichthys olivaceus) embryonic cells (FEC) after treatment with UV-inactivated turbot (Scophthalmus maximus L.) rhabdovirus (SMRV), termed PoHsp40A4, PoHsp40B6 and PoHsp40B11, whose encoded proteins all possess the conserved DnaJ domain, a signature motif of the Hsp40 family. Based on different protein structure and phylogenetic analysis, they can be categorized into two subfamilies, PoHsp40A4 for Type I Hsp40, PoHsp40B6 and PoHsp40B11 for Type 11 Hsp40. Further expression analysis revealed two very different types of kinetics in response either to heat shock or to virus infection, with a marked induction for PoHsp4OA4 and a weak one for both PoHsp40B6 and PoHsp40B11. A very distinct tissue distribution of mRNA was also revealed among the three genes, even between PoHsp40B6 and PoHsp40B11. This is the first report on the transcriptional induction of Hsp40 in virally stimulated fish cells, and the differential expressions might reflect their different roles in unstressed and stressed cells. (c) 2005 Elsevier Ltd. All rights reserved.

Responses of antioxidant system in Arabidopsis thaliana suspension cells to the toxicity of microcystin-RR

Microcystins are cyclic heptapeptide hepatoxins produced by many species of cyanobacteria. The toxic effects and mechanism of microcystins on animals have been well studied both in vivo and in vitro. It was also reported that microcystins had adverse effects on plants. However, to our knowledge, there is no information about the toxic effects and mechanism of microcystins on plant suspension cells. In this study, Arabidopsis thaliana suspension cells were exposed to a range dose of microcystin-RR. Lipid peroxidation, a main manifestation of oxidative damage, was studied and a time- and dose-dependent increase in malondiadehyde was observed. In contrast, glutathione (GSH) levels in the cells decreased after 48 h treatment with 1 and 5 mg/L of microcystin-RR. The activities of superoxide dismutase (SOD) and catalase (CAT) increased significantly after 48 h exposure to I and 5 mg/L of microcystin-RR, but glutathione S-transferase (GST) activity showed no difference compared with the control. These results clearly indicate that microcystin-RR is able to cause oxidative damage in A. thaliana suspension cells. Decrease of GSH content and increases of SOD and CAT activities reveal that the antioxidant system may play an important role in eliminating or alleviating the toxicity of microcystin-RR. The possible toxicity mechanism of microcystin-RR on the A. thaliana suspension cells is also discussed in this paper. (C) 2005 Elsevier Ltd. All rights reserved.

Humoral immune responses of the grouper Eninephelus akaara against the microsporidium Glugea epinephelusis

The humoral immune responses of grouper Epinephelus akaara to a natural infection with Glugea epinephelusis was studied by ELISA utilizing intact mature spores as the coated antigen. Results showed that a specific humoral immune response was elicited, but the intensity of infection (in terms of the number of cysts) was not related to the antibody level in naturally infected hosts. The differences in the antigenicity of intact mature spores and soluble spore proteins derived from cracked mature spores were also analyzed. Results suggested that similar antigen epitopes existed between the 2 groups. Additionally, antigen component patterns and the distribution of antigen with immunogenicity were investigated by using the western blot and the immunofluorescent antibody technique (IFAT). The new parasitic microsporidium has specific polypeptide patterns comparable to the reported fish microsporidians. The main antigenic substances are concentrated on the surface of spores, and are mostly located on the anterior and posterior end of the spore bodies. Most surface components of the G. epinephelusis spores are soluble, The potential role of the surface components in initiating infection was also discussed.

Biomarker responses and reproductive toxicity of the effluent from a Chinese large sewage treatment plant in Japanese medaka (Oryzias latipes)

The present study was conducted to assess the potential toxicity of the effluent from a large sewage treatment plant (GBD-STP) in Beijing. Japanese medakas (Oryzias latipes) at reproduction active period were exposed to a serial of graded concentrations of the effluent or 100 ng l(-1) of 17-alpha-ethinylestradiol (EE2, positive control). Growth, gonadosomatic index (GSI), hepatosomatic index (HSI), reproductive success, induction potency of vitellogenin (VTG) in male fish and that of 7-ethoxyresorufin-o-deethylase activity (EROD) in male fish liver were used as test endpoints. The growth suppression of fish was observed in a dose-dependent manner, resulting in significant differences in both body length and body weight of medaka above 5% effluent. This effluent can inhibit the growth of gonad of medakas and are more sensitive to male than to female. At exposure concentration of 40% and higher, there was an unexpected decrease of HSI values, which may be resulted from sub-lethal toxicity of effluent to fish liver. VTG of plasma in males were induced in all exposure concentration levels, but not in a dose-dependent manner. The concentration of 5% effluent would be the lowest observed adverse effect level (LOAEL) affecting reproductive success when examining fertile individuals, fecundity and fertilization rate. The overt CYP1A response and higher reproductive toxicity may be indicative of low process efficiency of this STP. (c) 2004 Elsevier Ltd. All rights reserved.

Acute biochemical responses of a submersed macrophyte, Potamogeton crispus L., to high ammonium in an aquarium experiment

Acute biochemical responses of Potamogeton crispus L. to high external ammonium were investigated in an aquarium experiment. Shoots of P. crispus were incubated in aquaria for 24 h or 48 h at five treatments of ammonium-0, 1, 5, 10 and 20 mg/L NH4-N. Soluble sugar content of the shoots declined markedly with increasing ammonium levels, whereas soluble amino acid content increased dramatically. Responses of two antioxidant enzymes as well as soluble protein content fit a lognormal distribution with increasing ammonium levels. High ammonium levels (NH4-N greater than or equal to 5 mg/L) caused significant acute biochemical changes in P. crispus, which potentially could lead to significant biochemical damage.

Responses of antioxidant systems in the hepatocytes of common carp (Cyprinus carpio L.) to the toxicity of microcystin-LR

The freshwater, bloom-forming cyanobacterium (blue-green alga) Microcystis aeruginosa produces a peptide hepatotoxin, which causes the damage of animal liver. Recently, toxic Microcystis blooms frequently occur in the eutrophic Dianchi Lake (300 km(2) and located in the South-Westem of China). Microcystin-LR from Microcystis in Dianchi was isolated and purified by high performance liquid chromatography (HPLC) and its toxicity to mouse and fish liver was studied (Li et al., 2001). In this study, six biochemical parameters (reactive oxygen species, glutathione, superoxide dismutase, catalase, glutathione peroxide and glutathione S-transferase) were determined in common carp hepatocytes when the cells were exposed to 10 mug microcystin-LR per litre. The results showed that reactive oxygen species (ROS) contents increased by more than one-time compared with the control after 6 h exposure to the toxin. In contrast, glutathione (GSH) levels in the hepatocytes exposed to microcystin-LR decreased by 47% compared with the control. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxide (GSH-Px) increased significantly after 6 h exposure to microcystin-LR, but glutathione S-transferase (GST) activity showed no difference from the control. These results suggested that the toxicity of microcystin-LR caused the increase of ROS contents and the depletion of GSH in hepatocytes exposed to the toxin and these changes led to oxidant shock in hepatocytes. Increases of SOD, CAT and GSH-Px activities revealed that these three kinds of antioxidant enzymes might play important roles in eliminating the excessive ROS. This paper also examined the possible toxicity mechanism of microcystin-LR on the fish hepatocytes and the results were similar to those with mouse hepatocytes. (C) 2003 Elsevier Science Ltd. All rights reserved.

Photosynthetic responses to inorganic carbon in Ulva lactuca under aquatic and aerial states

Intertidal macroalgae experience continual alternation of photosynthesis between aquatic state at high tide and aerial state at low tide. The comparative photosynthetic responses to inorganic carbon were investigated in the common intertidal macroalga Ulva lactuca L. along the coast of Shantou between aquatic and aerial state. The inorganic carbon dissolved in seawater at present could fully (at 10 degreesC or 20 degreesC) or nearly (at 30 degreesC) saturate the aquatic photosynthesis of U. lactuca. However, the aerial photosynthesis was limited by current ambient atmospheric CO2 level, and such a limitation was more severe at higher temperature (20degrees - 30degrees T) than at lower temperature (10 T). The carbon-saturated maximal photosynthesis of U. lactuca under aerial state was much greater than that under aquatic state at 10 degreesC and 20 degreesC, while the maximal photosynthesis under both states was similar at 30 degreesC. The aerial values of K-m (CO2) for photosynthesis were higher than the aquatic values. On the contrary, the values of apparent photosynthetic CO2 conductance under aerial state were considerably lower than that under aquatic state. It was concluded that the increase of atmospheric CO2 would enhance the primary productivity of U. lactuca through stimulating the photosynthesis under aerial state during low tide.

Effects of CO2 enrichment on the bloom-forming cyanobacterium Microcystis aeruginosa (Cyanophyceae): Physiological responses and relationships with the availability of dissolved inorganic carbon

Microcystis aeruginosa Kutz. 7820 was cultured at 350 and 700 muL.L-1 CO2 to assess the impacts of doubled atmospheric CO2 concentration on this bloom-forming cyanobacterium. Doubling Of CO2 concentration in the airflow enhanced its growth by 52%-77%, with pH values decreased and dissolved inorganic carbon (DIC) increased in the medium. Photosynthetic efficiencies and dark respiratory rates expressed per unit chl a tended to increase with the doubling of CO2. However, saturating irradiances for photosynthesis and light-saturated photosynthetic rates normalized to cell number tended to decrease with the increase of DIC in the medium. Doubling of CO2 concentration in the airflow had less effect on DIC-saturated photosynthetic rates and apparent photosynthetic affinities for DIC. In the exponential phase, CO2 and HCO3- levels in the medium were higher than those required to saturate photosynthesis. Cultures with surface aeration were DIC limited in the stationary phase. The rate of CO2 dissolution into the liquid increased proportionally when CO2 in air was raised from 350 to 700 muL.L-1, thus increasing the availability of DIC in the medium and enhancing the rate of photosynthesis. Doubled CO2 could enhance CO2 dissolution, lower pH values, and influence the ionization fractions of various DIC species even when the photosynthesis was not DIC limited. Consequently, HCO3- concentrations in cultures were significantly higher than in controls, and the photosynthetic energy cost for the operation of CO2 concentrating mechanism might decrease.

Comparison of compensatory growth responses of juvenile three-spined stickleback and minnow following similar food deprivation protocols

The compensatory growth responses of individual juveniles of two co-existing species were compared after identical periods of starvation to determine inter-specific similarities and differences. The carnivorous stickleback Gasterosteus aculeatus was compared with the omnivorous minnow Phoxinus phoxinus. Both species experienced 1 or 2 weeks of starvation before being re-fed ad libitum. The two species differed in their response to the starvation periods, with minnows showing a lower weight-specific loss. Both species showed compensatory responses in appetite, growth and to a lesser extent, growth efficiency. Minnows wholly compensated for 1 and 2 weeks of starvation. At the end of the experiment, sticklebacks starved For 2 weeks were still showing a compensatory response and had nut achieved full compensation. The compensatory responses of the sticklebacks showed a lag of a week before developing in the re-feeding phase, whereas the response of the minnows was immediate. Analysis of lipid and dry matter concentrations suggested that the compensatory response restored reserve lipids while also bringing the fish back to the growth trajectory of continuously fed fish. (C) 2001 The Fisheries Society of the British Isles.

Cell responses of Dunaliella salina FACHB 435 (green alga) to microgravitational stimulation by clinorotation

Clinorotation experiments were established to simulate microgravity on ground. It was found that there were obvious changes of Dunaliella salina FACHB435 cells and their metabolic characteristics during clinorotation. The changes included the increases of glycerol content, the rate of H+ secretion and PM H+-ATPase activity, and the decrease of ratio of the plasma membrane (PM) phospholipid to PM protein. These results indicated that microgravity was a stress environment to Dunaliella salina. It is deduced that it would be possible to attribute the effect of microgravity on algal cells to the secondary activation of water stress.

IMPROVED PEAK POWER METHOD FOR MEASURING FREQUENCY RESPONSES OF PHOTODETECTORS IN A SELF-HETERODYNE SYSTEM

An improved peak power method for measuring frequency responses of photodetectors in a self-heterodyne system consisting of a distributed Bragg reflector laser is proposed. The time-resolved spectrum technique is used to measure the peak power of the beat signal and the intrinsic linewidth of heat signal for calibration. The experimental results show that the impact of the thermal-induced frequency drift, which is the main reason for producing an error in measurement by conventional peak power method and spectrum power method, can be removed.

Subtraction of scattering parameters for adiabatic intrinsic responses of semiconductor lasers

Thermal effects will make chip temperature change with bias current of semiconductor lasers, which results in inaccurate intrinsic response by the conventional subtraction method. In this article, an extended subtraction method of scattering parameters for characterizing adiabatic responses of laser diode is proposed. The pulsed injection operation is used to determine the chip temperature of packaged semiconductor laser, and an optimal injection condition is obtained by investigating the dependence of the lasing wavelength on the width and period of the injection pulse in a relatively wide temperature range. In this case, the scattering parameters of laser diode are measured on adiabatic condition and the adiabatic intrinsic responses of packaged laser diode are first extracted. It is found that the adiabatic intrinsic responses are evidently superior to those without thermal consideration. The analysis results indicate that inclusion of thermal. effects is necessary to acquire accurate intrinsic responses of semiconductor lasers. (C) 2008 Wiley Periodicals, Inc.

Measurement of small-signal and large-signal responses of packaged laser modules at high temperature

In this paper, the pulsed injection method is extended to measure the chip temperature of various packaged laser modules, such as the DFB laser modules, the FP laser modules, and the EML laser modules. An optimal injection condition is obtained by investigating the dependence of the lasing wavelength on the width and period of the injection pulse in a relatively wide temperature range. The small-signal frequency responses and large-signal performances of packaged laser modules at different chip temperature are measured. The adiabatic small-signal modulation characteristics of packaged LD are first extracted. In the large-signal measurement, the effects of chip temperature, bias current and driving signal on the performances of the laser modules are discussed. It has been found that the large-signal performances of the EML modules depend on the different red-shift speeds of the DFB and EAM sections as chip temperature varying, and the optimal characteristics may be achieved at higher temperature.

Improved optical heterodyne methods for measuring. frequency responses of photodetectors

An improved optical self-heterodyne method utilizing a distributed Bragg reflector (DBR) tunable laser and an optical fiber ring interferometer is presented in this paper. The interference efficiency can be increased by 7 dB compared with the scheme using the conventional Mach-Zehnder interferometer. The unsteady process that the beating frequency experiences in each tuning period is investigated. According to the measurement results, the wavelength and optical power of the tunable laser will be steady when the square-wave frequency is lower than 300 kHz. It has been shown that when a square-wave voltage is applied to the phase section of the tunable laser, the laser linewidths vary in a wide range, and are much larger than that under dc voltage tuning. The errors caused by the variations in the linewidth of the beat signal and optical power can be eliminated using the proposed calibration procedures, and the measurement accuracy can, therefore, be significantly improved. Experiments show that the frequency responses obtained using our method agree well with the data provided by the manufacturer, and the improved optical self-heterodyne method is as accurate as the intensity noise technique.