Evaluation of copper-induced DNA damage in Vitis vinifera L. using Comet-FISH

The contamination of soils and water with copper (Cu) can compromise the crops production and quality. Fungicides containing Cu are widely and intensively used in viticulture contributing to environmental contamination and genotoxicity in Vitis vinifera L. Despite the difficulty in reproducing field conditions in the laboratory, hydroponic solutions enriched with Cu (1, 10, 25 and 50 μM) were used in forced V. vinifera cuttings to evaluate the DNA damage in leaves of four wine-producing varieties (‘Tinta Barroca’, ‘Tinto Cão’, ‘Malvasia Fina’ and ‘Viosinho’). Alkaline comet assay followed by fluorescence in situ hybridisation (Comet-FISH) was performed with the 45S ribosomal DNA (rDNA) and telomeric [(TTTAGGG)_n] sequences as probes. This study aimed to evaluate the tolerance of the four varieties to different concentrations of Cu and to determine which genomic regions were more prone to DNA damage. The comet assay revealed comets of categories 0 to 4 in all varieties. The DNA damage increased significantly (p < 0.001) with the Cu concentration. ‘Tinto Cão’ appeared to be the most sensitive variety because it had the highest DNA damage increase in 50 μM Cu relative to the control. Comet-FISH was only performed on slides of the control and 50 μM Cu treatments. Comets of all varieties treated with 50 μM Cu showed rDNA hybridisation on the head, ‘halo’ and tail (category III), and their frequency was significantly higher than that of control. The frequency of category III comets hybridised with the telomeric probe was only significantly different from the control in ‘Malvasia Fina’ and ‘Tinta Barroca’. Comet-FISH revealed partial damage on rDNA and telomeric DNA in response to Cu but also in control, confirming the high sensitivity of these genomic regions to DNA fragmentation.

     

Variation in some chemical parameters and organic matter in soils regenerated by the addition of municipal solid waste

The organic fraction of a municipal solid waste was added in different doses to an eroded soil formed of loam and with no vegetal cover. After three years, the changes in macronutrient content and the chemical-structural composition of its organic matter were studied. The addition of the organic fraction from a municipal solid waste had a positive effect on soil regeneration, the treated soils being covered with spontaneous vegetation from 1 yr onwards. An increase in electrical conductivity and a fall in pH were noted in the treated soils as were increases in macronutrients, particularly N and available P and the different carbon fractions. Optical density measurements of the organic matter extracted with sodium pyrophosphate showed that the treated soils contained an organic matter with less condensed compounds and with a greater tendency to evolve than the control. A pyrolysis-gas chromatography study of the organic matter extracted with pyrophosphate showed large quantities of benzene both in the treated soils and control; pyrrole was also relatively abundant, although this fragment decreased as the dose rose. Xylenes and pyridine were present in greater quantities in the control and furfural in the treated soils. Three years after addition to the soil, the organic matter had a higher proportion of fragments derived from aromatic compounds and a smaller proportion derived from hydrocarbons. Similarity indices showed that, although the added and newly formed organic matter 3 yr after addition continued to differ from that of the original soil and to be more mineralizable, the transformations it has undergone made it more similar to the original organic matter of the soil than it was at the moment of being added.     

Prenatal diagnosis of thalassemia major by fetal blood analysis: Experience with 1000 cases

In this report we have summarized our experience with the prenatal diagnosis of β-thalassemia in 1000 pregnancies followed at least until 12 months after birth. In the majority of these cases, the thalassemia lesion was the nonsense mutation at the codon corresponding to amino acid 39, which produces the hematological phenotype of β-thalassemia. Fetal blood sampling was carried out by placental aspiration, by which a sufficient amount of fetal blood for analysis was obtained in the majority of cases (99 per cent). The fetal mortality associated with fetal blood sampling was 6·3 per cent. Those placental samples contaminated by maternal cells were successfully purified by Ørskov lysis. Fetal blood was analysed by globin chain synthesis on CM–52 columns, which gave reliable results. Two misdiagnoses (0·2 per cent) have been made of which one was due to a non-globin protein co-migrating with the β-chains while the other resulted from a misclassification of the type of thalassemia segregating in the family.     

Local changes in the composition and community structure of suprabenthic peracarid crustaceans on the bathyal Mediterranean: influence of environmental factors

Deep-water assemblages of suprabenthic peracarids were analyzed in the SW Balearic Islands (Algerian Basin, southwestern Mediterranean) between 249 and 1,622 m depth; the patterns of species composition, possible zonation, and trophic structure found in this area were compared with those exhibited by peracarids in the mainland side of the Catalan Sea slope (northwestern Mediterranean). One hundred and four peracarid species (plus one leptostracan) were identified on the Balearic Islands slope, amphipods being the most diversified taxon (45 species). On the Balearic slope, two distinct depth assemblages were distinguished: one at the upper slope (US), between 249 and 402 m depth and the second at the deep slope, between 543 and 1,620 m depth. A remarkable species substitution occurred at depths between 402 and 638 m. In the Catalan Sea, in addition to the US assemblage occupying depths between 208 and 408 m, a second boundary of faunal change was found around 1,250 m. Suprabenthos biomass increased from 242 to approximately 500 m. Suprabenthos attained the highest biomass values (100 g wet weight/10,000 m²) at intermediate depths between 504 and 1,211 m, as also occurred with the associated zooplankton collected with suprabenthos (peak biomass between 502 m and 898 m). Suprabenthos biomass did not show any significant correlation with any environmental water-column variable. In contrast, zooplankton (especially small fish and decapod crustaceans) showed a significant positive correlation with fluorometry and turbidity at different levels of the water column. The feeding guilds of species showed important differences between the two areas only on the US, with a higher abundance of deposit feeders in the Catalan Sea (20.4%) than in the Balearic Islands (4.2%). The low contribution of deposit feeders in the SW Balearic Islands may ultimately be a consequence of the lack of river discharges in this area.Communicated by S.A. Poulet, Roscoff     

Early life histories in the bryozoan<Emphasis Type="Italic"> Schizobrachiella sanguinea</Emphasis>: a case study

Mediterranean populations of Schizobrachiella sanguinea seem to span a wide range of contrasting benthic habitats. We have studied adult, larval, and recruit populations of S. sanguinea from sublittoral (approximately 10 m depth) communities at Blanes (Spain, NW Mediterranean). At the study site, the species colonises semi-obscure caves and, to a lesser extent, communities of sciaphilic algae. Our first goal was to show the periods of brooding, larval release, and recruitment. Plankton was collected about twice a week between July 1998 and June 2000 over an artificial reef. A total of 303 larvae were collected in 23 of the 102 days of sampling. At the study site the species shows a single, annual larval release period, which occurs from late March to June–July every year. Recruits were found from late May to late August 1999 and in late May 2000. A recruitment peak was observed in July 1999 and again in late May 2000. The presence of brooding adults corresponded to the periods of larval supply and recruitment. Our second goal was to investigate the causes of the variable distribution of the species along several communities in the north-west Mediterranean Sea. We describe adult distribution and the dynamics of larval supply, recruitment of early (15 days from settlement) and late (4 months from settlement) recruits. Patterns of larval presence and early recruit distribution along the communities on the reef did not reflect the adult distributions, whereas this was closely matched by the distribution of late recruits. Post-recruitment mortality strongly affected recruits of the PA (photophilic algae) and SA (sciaphilic algae) communities, whereas it was very low in the communities of SOC (semi-obscure caves). Mortality of recruits was density independent and colony survivorship in the SA communities, where recruitment was the highest, appeared strongly affected by competition with brown algae. Like the majority of cheilostomate bryozoans, S. sanguinea releases a coronate larva capable of active swimming and positive reaction to light. Such swimming abilities may allow larvae to actively select the substrate on which to settle and to avoid unsuitable substrates. It seems that larval behaviour does not explain the preference of S. sanguinea for sciaphilic habitats. High post-recruitment mortality due to various factors, especially competition with fast-growing algae, seems particularly relevant in determining the adult distributions observed.Communicated by S.A. Poulet, Roscoff     

The Role of a Salt Marsh Plant on Trace Metal Bioavailability in Sediments. Estimation By Different Chemical Approaches * (7 pp)

Goal, Scope and Background The presence or absence of vegetation can condition sediment characteristics. The main aim of this work was to investigate the influence of the sea rush Juncus maritimus on metal (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) availability to organisms living on or in estuarine sediments, from Douro River (NW Portugal), by comparing the characteristics and chemical behaviour of rhizosediments (collected within the plant assemblage) and those of sediment (collected around the plant). In order to evaluate whether and how sediment characteristics condition the role of plants on metal availability, sandy and muddy sediments colonised by J. maritimus were studied in parallel. Methods Metal availability was estimated by enzymatic digestion with pepsin (ED), which may provide an estimate of metal availability to organisms living at estuarine sediments. Nevertheless, since no consensus exists yet on the most suitable methodologies to estimate metal bioavailability in sediments, two more conventional approaches, BCR sequential extraction (SE) and AVS/SEM model, were also used, in parallel, and the information these approaches provided was compared with that provided by ED. Total-recoverable metal contents were determined by atomic absorption spectrophotometry after sediment digestion using a high-pressure microwave system. Results and Discussion Plants could concentrate metals around its roots and rhizomes. In addition, they were capable of oxidizing (release of oxygen by the roots) the anaerobic medium surrounding their roots in muddy sediment (reducing AVS). As sulphide oxidation renders metals (Cd, Cu, Ni, Pb and Zn) into more soluble forms, according to the AVS/SEM model, metals from muddy sites would be more available in rhizosediment than in sediment. The SE approach led to a similar conclusion. Nevertheless, the results provided by ED pointed at opposite conclusion, particularly for Cd and Zn, indicating less availability at rhizosediments than in the surrounding sediment. ED results were interpreted as a consequence of an enrichment of the rhizosediment in organic ligands exuded by the roots or liberated by dead plants. The effect of complexation of metals by organic compounds, which ED could not decompose/dissolve, seemed to overcome that caused by sediment oxidation. In general, a comparison of the information about metal availability by ED, SE, AVS/SEM, showed that it did not always match and in few cases it was even contradictory. Conclusion and Outlook Therefore, a thorough evaluation of the metal availability in sediments requires a combination of different chemical approaches, so as to take into consideration differences in ways of organism exposure (interstitial water and/or ingestion of sediment particles). - * The basis of this peer-reviewed paper is a presentation at the 9th FECS Conference on 'Chemistry and Environment', 29 August to 1 September 2004, Bordeaux, France.     

Study of metal fractionation in river sediments. A comparison between kinetic and sequential extraction procedures

The extraction kinetic of trace metals (Cd, Cu, Pb, and Zn) in river sediments by four extraction agents was studied. As extractants ammonium acetate, acetic acid, hydroxylamine and EDTA solutions were assayed. These reagents can leach the metals more or less selectively from several metal compartments of sediments. The metal leaching kinetic model permits classification of the metal species in labile and moderately-labile ones. The combination of two or more non specific reagents permits a high characterisation of metal distribution and leachability. The results obtained with this model in four river sediments were compared with data obtained by the SM&T sequential extraction procedure, in order to characterise the chemical nature of leached metal.     

Effect of alkalinity on the performance of a simulated landfill bioreactor digesting organic solid wastes

This study investigated the effects of alkalinity on the anaerobic treatment of the organic solid wastes collected from the kitchen of Engineering Faculty in Dokuz Eylul University, Izmir, Turkey and the leachate characteristics treated in three simulated landfill anaerobic bioreactors. All of the reactors were operated with leachate recirculation. One reactor was operated without alkalinity addition. The second reactor was operated by the addition of 3 g l-1 d-1 of NaHCO3 alkalinity to the leachate and the third reactor was operated by the addition of 6 g l-1 d-1 NaHCO3 alkalinity to the leachate. After 65 d of anaerobic incubation, it was observed that the chemical oxygen demand (COD), volatile fatty acids (VFA) concentrations, and biochemical oxygen demand to chemical oxygen demand (BOD5/COD) ratios in the leachate samples produced from the alkalinity added reactors were lower than the control reactor while the pH values were higher than the control reactor. The COD values were measured as 18900, 3800 and 2900 mg l-1 while the VFA concentrations were 6900, 1400 and 1290 mg l-1, respectively, in the leachate samples of the control, and reactors containing 3 g l-1 NaHCO3 and 6 g l-1 NaHCO3 after 65 d of anaerobic incubation. The total nitrogen (TN), total phosphorus (TP) and ammonium nitrogen (NH4-N) concentrations in organic solid waste (OSW) significantly reduced in the reactor containing 6 g l-1 NaHCO3 by d 65. The values of pH were 6.54, 7.19 and 7.31, after 65 d of anaerobic incubation, respectively, in the aforementioned reactors results in neutral environmental conditions in alkalinity added reactors. Methane percentage of the control, reactors containing 3 g l-1 NaHCO3 and 6 g l-1 NaHCO3 were 37%, 64% and 65%, respectively, after 65 d of incubation. BOD5/COD ratios of 0.27 and 0.25 were achieved in the 3 and 6 g l-1 NaHCO3 containing reactors, indicating a better OSW stabilization. Alkalinity addition reduced the waste quantity, the organic content of the solid waste and the biodegradation time.     

Spectrophotometric determination of the fungicide captan

A spectrophotometric method for the determination of the fungicide captan has been developed based on its reaction with thiosemicarbazide. The absorbance was measured at 315 nm. The effects of the proportion of water, thiosemicarbazide concentration, pH and temperature on this reaction was studied to select the best chemical conditions. The calibration graph was obtained between 1.2 and 30 ppm and the detection limit is 0.5 ppm. The RSD (n = 10) was 1.34%. Possible interference of various pesticides was examined. The method was applied to the determination of captan in agrochemical formulations. Results agree well with the labelled values and also with results that were obtained by a polarographic method.