978 resultados para sybr green fluorescence assay


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order to investigate the morphological response of freshwater green algae to elevated CO2 concentration, Chlamydomonas reinhardtii Dang and Scenedesmus obliquus Kutz were cultured with enriched CO2, and their microstructure and ultrastructure were examined by microscopy and electron microscopy. The effect of CO2 enrichment to 186 mumol/L, was insignificant on the shape and size of C. reinhardtii, but significant in reducing the volume of S. obliquus. High-CO2 increased the amount of chloroplast. The pyrenoids occurred in low-CO2-grown cells but not in high-CO2-grown ones and more starch granules were observed in the former.

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Photosynthetic responses of rewetted Nostoc flagelliforme to CO2, desiccation, light and temperature were investigated under emersed conditions in order to characterize its ecophysiological behaviour in nature. Net photosynthesis increased to a maximum rate at about 30 % water loss, then decreased, while dark respiration always decreased with the progress of desiccation. Light-saturated photosynthesis and dark respiration were significantly reduced at 8 degreesC, but remained little affected by changes of temperature within the range of 15-35 degreesC. Photosynthetic efficiency (alpha) was maximal at the beginning of desiccation and then reduced with increased water loss. Saturating irradiance for photosynthesis was about 194-439 mu mol quanta m(-2) s(-1), being maximal at about 30 % water loss. No photoinhibition was observed at irradiances up to 1140 mu mol m(-2) s(-1). Light compensation points were about 41-93 mu mol m(-2) s(-1). Photosynthesis of N. flagelliforme was CO2-limited at the present atmospheric CO2 level. The CO2-saturated photosynthesis increased with increase of irradiance (190-1140 mu mol m(-2) s(-1)) and temperature (8-25 degreesC) and decreased significantly with water loss (0-75 %). Photosynthetic affinity for CO2 was sensitive to temperature and irradiance. The CO2 compensation point (Gamma) increased significantly with increased temperature and was insensitive to irradiance. Desiccation did not affect Gamma values before water loss exceeded 70 %. Photorespiratory CO2 release did not occur in N. flagelliforme at the current atmospheric CO2 level.

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Green-lipped mussels (Perna viridis) were collected from a site in Hong Kong which is relatively free from polycyclic aromatic hydrocarbon (PAH) contamination, and maintained in situ at this and three other sites with different degrees of PAH contamination. The transplanted mussels were retrieved after a 30-day field exposure. DNA adducts in the gill tissues were quantified, and tissue concentrations of benzo[a]pyrene as well as total PAHs (with potential carcinogenicity) determined for individual mussels. Results indicate that (1) tissue concentration of PAHs and adduct levels in mussels collected from a single site can be highly variable; and (2) adduct levels were related to tissue concentrations of benzo[a]pyrene as well as total PAHs of individual animals.

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A new Enzyme ImmunoAssay (EIA) for PCDD/F TEQ measurement in extracts of environmental samples was described. The bioassay TEQ which derived from EIA and EROD were compared with each other and with results from chemical analysis. For all environmental samples, the EROD-TEQ is higher than the value from chemical analysis. However, the EIA-TEQ is much more identical with the value from chemical analysis. Our results indicate that the EIA assay is a complementary method to the EROD assay and should be useful as a rapid and sensitive screening tool for environmental samples in many situations. (C) 1999 Elsevier Science Ltd. All rights reserved

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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.

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Nostoc flagelliforme, which is distributed in arid or semiarid steppes of the west and west-northern parts of China, has been used by the Chinese as a food delicacy and for its herbal values for hundreds of years. However, the resource is being over-exploited and is diminishing, while the market demands are increasing with the economic growth. This review deals mainly with the Chinese studies on the ecology, physiology, reproduction, morphology and culture of this species in an attempt to promote research and development of its cultivation technology.

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A settlement inhibition assay using barnacle cyprid larvae, Balanus amphitrite, was developed with Cd2+ and phenol as standard reference toxicants. Mean percentage settlement of cyprid larvae showed a progressive reduction with increasing concentrations of Cd2+ and phenol. A significant reduction in settlement was found when cyprids were exposed to 0.1 mgL(-1) Cd2+ or 10 mgL(-1) phenol. The assay was used to assess the sublethal toxicity of three oil dispersants (Vecom B-1425 GL, Norchem OSD-570 and Corexit 9905) commonly used in Hong Kong waters. Results of this investigation show that the barnacle settlement inhibition assay can be incorporated into the battery of tests currently available for ecotoxicological assessment of marine contaminants. (C) 1997 Elsevier Science Ltd.

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The effects of sublethal concentrations of phenol and cadmium on the phototactic responses of the stage II nauplii of the barnacle Balanus amphitrite were investigated. Increased toxicant concentrations caused a reduction in phototactic responses. Balanus amphitrite nauplii exposed to nominal phenol concentrations of 100 ppm and higher for 1-12 h failed to exhibit phototactic responses, while longer exposure times of 24 and 48 h reduced the lowest observable effect concentration (LOECs) to 80 and 60 ppm, respectively. For cadmium, the LOECs, based on nominal concentrations, for B. amphitrite following 1, 6, 12, 24, and 48 h exposures were 20, 4.5, 4.0, 1, and 0.75 ppm, respectively. The LOECs can be significantly reduced by increasing the duration of exposure to the toxicants. A good relationship exists between the phototactic response and toxicant concentration as well as exposure time. Results of this study indicate that the toxicant-induced reduction in phototactic responses of barnacle larvae can be used in a sensitive, rapid screening test for ecotoxicological assessments. (C) 1997 by John Wiley & Sons, Inc.

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Studies on mixed mass cultivation of Anabaena spp. on a large scale (5170 m2) were conducted continuously for 3 years. Under the continental monsoon climate in northern subtropics (30-degrees-N, 115-degrees-E), 7-11 g dry weight m-2 day-1 of microalgal biomass on average was harvested in simple plastic greenhouses in the effective growth days during the warmer seasons. The maximum productivity was 22 g m-2 day-1 in the middle of summer. Observations on the productive properties of strains of Anabaena spp. indicated that they were different from and could compensate for each other in their productivities and adaptations to the seasonal changes. With different lining materials (PVC sheets, concrete, sand and soil) in the culture ponds, no significant variation of productivity was found, but bubbling with biogas in the middle of the day and the application of some growth regulating substances (2,4-D, NaHSO3 and extracts of oyster mushroom spawn) was able to improve the production. The cost of microalgal biomass in this way was around 0.75-1.0 US dollar(s) per kilogram.

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Toxic cyanobacteria (blue-green algae) waterblooms have been found in several Chinese water bodies since studies began there in 1984. Waterbloom samples for this study contained Anabaena circinalis, Microcystis aeruginosa and Oscillatoria sp. Only those waterblooms dominated by Microcystis aeruginosa were toxic by the intraperitoneal (i.p.) mouse bioassay. Signs of poisoning were the same as with known hepatotoxic cyclic peptide microcystins. One toxic fraction was isolated from each Microcystis aeruginosa sample. Two hepatotoxic peptides were purified from each of the fractions by high-performance liquid chromatography and identified by amino acid analysis followed by low and high resolution fast-atom bombardment mass spectrometry (FAB-MS). LD50 i.p. mouse values for the two toxins were 245-mu-g/kg (Toxin A) and 53-mu-g/g (Toxin B). Toxin content in the cells was 0.03 to 3.95 mg/g (Toxin A) and 0.18 to 3.33 mg/kg (Toxin B). The amino acid composition of Toxin A was alanine [1], arginine [2], glutamic acid [1] and beta-methylaspartic acid [1]; for Toxin B it was the same, except one of the arginines was replaced with a leucine. Low- and high-resolution FAB-MS showed that the molecular weights were 1,037 m/z (Toxin A) and 994 m/z (Toxin B), with formulas of C49H76O12N13 (Toxin A) and C49H75O12N10 (Toxin B). It was concluded that Toxin A is microcystin-RR and Toxin B is microcystin-LR, both known cyclic heptapeptide hepatotoxins isolated from cyanobacteria in other parts of the world. Sodium borohydride reduction of microcystin-RR yielded dihydro-microcystin-RR (m/z = 1,039), an important intermediate in the preparation of tritium-labeled toxin for metabolism and fate studies.