A method for the assessment of light-induced oxidative stress in embryos of fucoid algae via confocal laserscan microscopy

A method was developed for measurement of active oxygen production in embryonic stages of the brown seaweed Fucus spiralis, using the label CM-DCFH-DA. Active oxygen species convert the label into the green fluorescent CM-DCF (exc/em 488/530 nm) that is detected via confocal laserscan microscopy and quantitative image analysis. Loading of the label did not harm the embryos; loading efficiency was age-independent, and the esterases needed for conversion to CM-DCFH were not inhibited by the effective UV dose (2 W m⁻²) applied in the experiments. After correction for daily variation of the laser power, and calibration with DCF standard solutions, this automated analysis of confocal images rendered active oxygen concentrations in fucoid embryos (μM DCF). An experiment was designed for the assessment of active oxygen production following irradiance stress in the light-sensitive embryos. Dim-light-acclimated, 1-, 2- and 4-day-old embryos were transferred for 60 min to defined high-light conditions (4π-irradiance 300 μmol photons m⁻² s⁻¹), optionally without UV radiation, including UVA, or including UVA plus UVB. PSII yield measurements (PAM fluorometer) were carried out in order to assess the degree of photoinhibition under these light conditions. The imposed light stress initially caused a rapid decrease of the PSII yields (Φ_P). With increasing embryo age, minimum Φ_P values attained under light stress remained higher. Consequently, electron transport rates (ETR) would increase with embryo age, i.e., with the development of their photosynthetic apparatus. Active oxygen production increased with ETR, and when UVB was included, relatively greater amounts of active oxygen were produced. A slow, second-phase decrease of Φ_P under light stress that was proportional to active oxygen production indicated that some photooxidative damage was caused, in particular during UVB exposure. Recovery from light stress was a rapid process in the absence of UVB; in such cases Φ_P was almost restored to the initial values within 60 min. The relative state of recovery of Φ_P was correlated with both the effective UV dose and active oxygen production rate (DCF). Recovery was slowest in embryos exposed for 60 min to an experimental UVB dose, which was representative of a situation at low tide, on a sunny day. The results suggest that active oxygen may cause an in situ inhibition of growth of the earliest life stages of F. spiralis. Received: 26 January 2000 / Accepted: 4 September 2000     

The response of sea scallop (Placopecten magellanicus) veligers to a weak thermocline in 9-m deep mesocosms

We examined the vertical distributions of scallop (Placopecten magellanicus) veligers in deep (0.6 m diameter, 9.5 m deep) polyethylene mesocosms from December 1991 to January 1992. In the mesocosms temperature stratification varied from 0 to 1.5 °C. Profiles of vertical distribution revealed several repeated patterns. Peaks in veliger numbers often appeared at the water surface and just above the thermocline. Higher density patches were seen below the surface peaks, and revealed the presence of bio-convective cells. Distribution away from these discontinuities was usually even. Distribution of veligers was affected by thermoclines above 1.0 °C. Responses to thermoclines varied with larval age and time of day, and 28 to 30 d veligers passed in both directions through a 1.5 °C thermocline. We conclude that larval behaviour is a major determinant of whether veligers pass through a thermocline. Kinematic viscosity may play a role in perception of temperature changes. Two potential consequences of such behaviour are (1) remaining in more productive upper water layers, where feeding opportunities are enhanced, and (2) increased horizontal transport in the region of the thermocline, which may enhance recruitment. Received: 15 May 1996 / Accepted: 11 May 2000     

Monitoring method for airborne glymes and its application in fuel exhaust emission measurement.

A monitoring method based on solvent extraction of adsorbed target glymes followed by gas chromatograph-mass spectrometry GC-MS analysis was developed for seven glymes, namely ethylene glycol dimethyl ether, propylene glycol dimethyl ether, ethylene glycol diethyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, triethylene glycol dimethyl ether, and tetraethylene glycol dimethyl ether. The best recoveries of target glymes were achieved when using a combination of sample collection medium of graphitised carbon black (GCB) with a solvent mixture of methylene chloride and methanol (95/5, v/v). Method detection limits ranged from 1.5 microg/m3 for diethylene glycol diethyl ether to 13.2 microg/m3 for ethylene glycol diethyl ether based on a sample volume of 3.4 1. Using this method, diethylene glycol dimethyl ether and diethylene glycol diethyl ether were detected and measured successfully in diluted vehicle exhausts in diesel fuel engine tests.     

Risk assessment of etofenprox (vectron) on non-target aquatic fauna compared with other pesticides used as Simulium larvicide in a tropical environment

Within the rotational scheme developed by the Programme to fight the resistance of Simulium damnosum to chemical larvicides, there was an operational gap at discharges between 5 and 70 m³ s⁻¹ for the treatment of rivers where resistance to organophosphates was present. The use of permethrin and carbosulfan was precluded because of risk of environmental impact and, Bacillus thuringiensis ser. H-14 treatments were not envisageable due to cost and logistics constraints. Among the possible complementary groups of larvicides tested, the pseudo-pyrethroids, held promise, because of a mode of action similar to that of pyrethroids, but along with a usually lower toxicity for fish. Etofenprox, one of the pseudo-pyrethroids tested, shows a global detachment of non-target insects in 24 h close to that of pyraclofos, an organo-phosphorus compound (27 against 23%). In laboratory conditions, six times the operational dose which is 0.03 mg l⁻¹ 10 min, is needed to cause 50% mortality of Caridina sp. (a small shrimps species) and 30 times this same dose for 95% mortality. For fish species, a safety margin of 400–800 times the operational dose is observed for Oreochromis niloticus and 200–400 times for Tilapia zillii.     

Storage and distribution of trace metals and spheroidal carbonaceous particles (SCPs) from atmospheric deposition in the catchment peats of Lochnagar, Scotland.

Lochnagar is a remote mountain lake to the south-east of the Cairngorm region in Scotland. Its catchment receives anthropogenic trace metals solely from atmospheric deposition. Ten peat cores were taken from the catchment and analysis confirmed that they have been contaminated by trace metals. The peats have an high affinity for trace metals and this results in metal accumulation in the surface peat layers. The formation of trace metal sulphides may also reduce remobilisation. In this way, trace metals derived from atmospheric deposition have been scavenged and accumulated. In contaminated peat layers, 77.4% Hg, 89.6% Pb, 93.4% Cu, 72.4% Zn and 86.5% Cd of the total stored are from anthropogenic sources. The accumulated trace metals in the peats can potentially influence the lake system through erosion. Spheroidal carbonaceous particle (SCP) profiles were used to date the peat cores. By referring to the SCP profiles in the peats and comparing these with the trace metal profiles in the lake sediments, the mobility of trace metals in the catchment peats is confirmed.     

Fluoride accumulation in pasture forages and soils following long-term applications of phosphorus fertilisers.

Ingestion of soils with high fluoride (F) concentration may cause chronic fluorosis in grazing animals. Analysis of New Zealand pasture soils with long-term phosphorus (P) fertilisation histories showed that total surface soil (0-75 mm depth) F concentration increased up to 217-454 mg kg-1 with P fertiliser application. One-third to two-thirds of F applied in fertilisers resides in the top 75 mm soil depth. Pasture forage accumulation of F was low, and therefore, F intake by grazing animals through pasture consumption is expected to be much lower than F intake by soil ingestion. Ten annual applications of single superphosphate (30 and 60 kg P ha-1 year-1) to a Pallic Soil (Aeric Fragiaqualf) significantly increased total F and labile F (0.01 M CaCl2 extract) concentrations to 200 and 120 mm depths, respectively, of the 300 mm depth investigated. The mobility of F in the soil profile was similar to two other elements, P and cadmium derived from the fertiliser.     

Sampling optimization, at site scale, in contamination monitoring with moss, pine and oak.

With the aim of optimizing protocols for sampling moss, pine and oak for biomonitoring of atmospheric contamination and also for inclusion in an Environmental Specimen Bank, 50 sampling units of each species were collected from the study area for individual analysis. Levels of Ca, Cu, Fe, Hg, Ni, and Zn in the plants were determined and the distributions of the concentrations studied. In moss samples, the concentrations of Cu, Ni and Zn, considered to be trace pollutants in this species, showed highly variable long-normal distributions; in pine and oak samples only Ni concentrations were log-normally distributed. In addition to analytical error, the two main source of error found to be associated with making a collective sample were: (1) not carrying out measurements on individual sampling units; and (2) the number of sampling units collected and the corresponding sources of variation (microspatial, age and interindividual). We recommend that a minimum of 30 sampling units are collected when contamination is suspected.     

Response of foodborne Salmonella spp. marker strains inoculated on egg shell surfaces to disinfectants in a commercial egg washer.

The objective of this study was to determine the effectiveness of an iodine based disinfectant (IBD, Iocide, Biomedical Development Corporation, San Antonio, TX) on Salmonella enteritidis and S. typhimurium inoculated on egg shell surfaces under simulated industry egg processing conditions with a commercial egg washer used as the sanitizer delivery system. Re-circulated egg washer water containing 1.40-2.85 g/l total dissolved solids was obtained from a commercial egg processing. Sanitizing treatments consisted of distilled deionized water (DDW), IBD, and chlorine (CL; 200 ppm). All treatments (DDW, IBD and CL) significantly (p < 0.05) decreased Salmonella spp. populations on the shell compared to dry (no spray) egg controls. However, efficacy of egg sanitizers appeared to be dependent on the level of total dissolved solids in the egg wash water.     

The mutagenic activity of thirty polycyclic aromatic hydrocarbons (PAH) and oxides in urban airborne particulates

Data are presented on the mutagenicity of an organic extract of a composite sample of urban air particulates and of thirty PAH compounds in such samples, including four quinone derivatives, isolated quantitatively by thin-layer chromatography and identified by fluorescence or other spectral techniques. Mutagenic activity was determined by the Ames assay, using histidine auxotrophs of Salmonella typhimurium strains TA98, TA100, TA1537 and TA1538. All compounds were dissolved in dimethyl sulfoxide (DMSO) which was the least toxic of eight organic solvents tested.The mutagenic activity of a benzene extract of suspended particulates from the air of Hamilton, Ontario was significantly greater with strain TA98 than with strain TA100, suggesting the presence of more frame-shift acting mutagens. The mutagenic response of this extract was similar with and without S-9 activation.Mutagenic tests on the 30 PAH compounds indicated that only the benzo(a)pyrene quinones were direct acting mutagens. All of the chemical compounds were tested with and without S-9 activation. The following PAH showed unequivocal mutagenic activity, with S-9 activation: benz(a)anthracene, benzo-(a)pyrene, benzo(ghi)perylene, benzo(rst)pentaphene, benzophenanthrene, chrysene, 1, 2, 3, 4-dibenzanthracene, 2, 3, 6, 7-dibenzanthracene, and 3-methyl cholanthrene. The quinones of 1, 6-; 3, 6-; and 6, 12-benzo(a)pyrenes showed weak mutagenic activity but 3, 6-benzo(a)pyrene elicited also a photodynamic response.