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Aims: Classification of European bog vegetation (Oxycocco-Sphagnetea class); identification of diagnostic species for the class and vegetation subgroups (orders and alliances); development of an expert system for automatic classification of vegetation plots; and production of distribution maps of the Oxycocco-Sphagnetea class and its alliances. Location: Europe. Methods: A data set of vegetation-plot records was compiled to include various bog types over most of the European continent. An unsupervised classification (beta-flexible linkage method, Sørensen distance measure) and detrended correspondence analysis (DCA) ordination were applied. Formal definitions of syntaxa based on species presence and covers, and respecting the results of the unsupervised classification, were developed and included in a classification expert system. Results: The Oxycocco-Sphagnetea class, its two orders (Sphagno-Ericetalia tetralicis and Sphagnetalia medii) and seven compositionally distinct alliances were formally defined. In addition to the syntaxa included in EuroVegChecklist, three new alliances were distinguished: Rubo chamaemori-Dicranion elongati (subarctic polygon and palsa mires); Erico mackaianae-Sphagnion papillosi (blanket bogs of the northwestern Iberian Peninsula); and Sphagno baltici-Trichophorion cespitosi (boreal bog lawns). The latter alliance is newly described in this article. Conclusions: This first pan-European formalized classification of European bog vegetation partially followed the system presented in EuroVegChecklist, but suggested three additional alliances. One covers palsa and polygon mires, one covers Iberian bogs with endemics and one fills the syntaxonomical gap for lawn microhabitats in boreal bogs. A classification expert system has been developed, which allows assignment of vegetation plots to the types described.

1. Evaluation of restoration activities is indispensable to assess the extent to which targets have been reached. Usually, the main goal of ecological restoration is to restore biodiversity and ecosystem functioning, but validation is often based on a single indicator, which may or may not cope with whole-ecosystem dynamics. Network analyses are, however, powerful tools, allowing to examine both the recovery of various biotic and abiotic properties and the integrated response at community and ecosystem level. 2. We used restoration sites where topsoil was removed from former intensively managed grassland and seeds were added. These sites were between 3 and 32 years old. We assessed how plants, soil biota, soil properties and correlation-based interactions between biotic communities and their abiotic environment developed over time and compared the results with (i) intensively managed (not restored), and (ii) well-preserved targeted semi-natural grasslands. 3. Plant, nematode, fungal and prokaryotic diversity and community structures of the restored grasslands revealed clear successional patterns and followed similar trajectories towards targeted semi-natural grasslands. All biotic communities reached targeted diversity levels no later than 18 years post-restoration. 4. Ecological networks of intensively managed and short-term (~4 years) restored grasslands were less tightly connected compared to those found in mid- and long-term (~18–30 years) restored and target grasslands. Restoration specifically enhanced interactions among biotic communities, but reduced interactions between biotic communities and their abiotic environment as well as interactions among abiotic properties in the short- and mid-term. 5. Synthesis and applications: Overall, our study demonstrated that topsoil removal and seed addition were successful in restoring diverse, tightly coupled and well-connected biotic communities above- and below-ground similar to those found in the semi-natural grasslands that were restoration targets. Network analyses proved to be powerful in examining the long-term re-establishment of functionally connected biotic communities in restored ecosystems. Thus, we provide an approach to holistically assess restoration activities by notably considering the complexity of ecosystems, much in contrast to most traditional approaches., Journal of Applied Ecology, 59 (9), ISSN:0021-8901, ISSN:1365-2664

This is a dataset of Ellenberg-type indicator values for European vascular plant species described in this article: Tichý L., Axmanová I., Dengler J., Guarino R., Jansen F., Midolo G., Nobis M.P., Van Meerbeek K., Aćić S., Attorre F., Bergmeier E., Biurrun I., Bonari G., Bruelheide H., Campos J.A., Čarni A., Chiarucci A., Ćuk M., Ćušterevska M., Didukh Y., Dítě D., Dítě Z., Dziuba T., Fanelli G., Fernández-Pascual E., Garbolino E., Gavilán R.G., Gégout J.-C., Graf U., Güler B., Hájek M., Hennekens S.M., Jandt U., Jašková A., Jiménez-Alfaro B., Julve P., Kambach S., Karger D.N., Karrer G., Kavgacı A., Knollová I., Kuzemko A., Küzmič F., Landucci F., Lengyel A., Lenoir J., Marcenò C., Moeslund J.E., Novák P., Pérez-Haase A., Peterka T., Pielech R., Pignatti A., Rašomavičius V., Rūsiņa S., Saatkamp A., Šilc U., Škvorc Ž., Theurillat J.-P., Wohlgemuth T. & Chytrý M. (2023) Ellenberg-type indicator values for European vascular plant species. Journal of Vegetation Science, 34, e13168. https://doi.org/10.1111/jvs.13168 The dataset contains: 1) Harmonized Ellenberg-type indicator values for light, temperature, moisture, reaction, nutrients, and salinity for European vascular plants. 2) Original indicator values from 13 datasets of Ellenberg-type indicator values: - Germany (Ellenberg & Leuschner 2010) - Austria (Karrer 1992) - Cantabrian Range (Jiménez-Alfaro et al. 2021) - Czech Republic (Chytrý et al. 2018) - European mires (Hájek et al. 2020) - France (Julve 2015) - Great Britain (Hill et al. 2000) - Greece (South Aegean) (Böhling et al. 2002) - Hungary (Borhidi 1995) - Italy (Guarino & La Rosa 2019, modified) - Saline habitats (Dítě et al. 2023) - Switzerland and the Alps (Landolt et al. 2010) - Ukraine (Didukh 2011) 3) Species nomenclature is standardized according to the Euro+Med PlantBase (http://europlusmed.org)., The final version of the paper (accepted 8th Dec 2022; published 22nd Dec 2022). Difference from version 1.0: Minor changes in newly added species values calculated from co-occurrence with other species in the EVA database. They were corrected using straight-line calibration.

It is generally assumed that restoring biodiversity will enhance diversity and ecosystem functioning. However, to date, it has rarely been evaluated whether and how restoration efforts manage to rebuild biodiversity and multiple ecosystem functions (ecosystem multifunctionality) simultaneously. Here, we quantified how three restoration methods of increasing intervention intensity (harvest only < topsoil removal < topsoil removal + propagule addition) affected grassland ecosystem multifunctionality 22 yr after the restoration event. We compared restored with intensively managed and targeted seminatural grasslands based on 13 biotic and abiotic, above‐ and belowground properties. We found that all three restoration methods improved ecosystem multifunctionality compared to intensively managed grasslands and developed toward the targeted seminatural grasslands. However, whereas higher levels of intervention intensity reached ecosystem multifunctionality of targeted seminatural grasslands after 22 yr, lower intervention missed this target. Moreover, we found that topsoil removal with and without seed addition accelerated the recovery of biotic and aboveground properties, and we found no negative long‐term effects on abiotic or belowground properties despite removing the top layer of the soil. We also evaluated which ecosystem properties were the best indicators for restoration success in terms of accuracy and cost efficiency. Overall, we demonstrated that low‐cost measures explained relatively more variation of ecosystem multifunctionality compared to high‐cost measures. Plant species richness was the most accurate individual property in describing ecosystem multifunctionality, as it accounted for 54% of ecosystem multifunctionality at only 4% of the costs of our comprehensive multifunctionality approach. Plant species richness is the property that typically is used in restoration monitoring by conservation agencies. Vegetation structure, soil carbon storage and water‐holding capacity together explained 70% of ecosystem multifunctionality at only twice the costs (8%) of plant species richness, which is, in our opinion, worth considering in future restoration monitoring projects. Hence, our findings provide a guideline for land managers how they could obtain an accurate estimate of aboveground‐belowground ecosystem multifunctionality and restoration success in a highly cost‐efficient way. © 2020 by the Ecological Society of America ISSN:1051-0761 ISSN:1939-5582

Aim The EUNIS Habitat Classification is a widely used reference framework for European habitat types (habitats), but it lacks formal definitions of individual habitats that would enable their unequivocal identification. Our goal was to develop a tool for assigning vegetation-plot records to the habitats of the EUNIS system, use it to classify a European vegetation-plot database, and compile statistically-derived characteristic species combinations and distribution maps for these habitats. Location Europe. Methods We developed the classification expert system EUNIS-ESy, which contains definitions of individual EUNIS habitats based on their species composition and geographic location. Each habitat was formally defined as a formula in a computer language combining algebraic and set-theoretic concepts with formal logical operators. We applied this expert system to classify 1,261,373 vegetation plots from the European Vegetation Archive (EVA) and other databases. Then we determined diagnostic, constant and dominant species for each habitat by calculating species-to-habitat fidelity and constancy (occurrence frequency) in the classified data set. Finally, we mapped the plot locations for each habitat. Results Formal definitions were developed for 199 habitats at Level 3 of the EUNIS hierarchy, including 25 coastal, 18 wetland, 55 grassland, 43 shrubland, 46 forest and 12 man-made habitats. The expert system classified 1,125,121 vegetation plots to these habitat groups and 73,188 to other habitats, while 63,064 plots remained unclassified or were classified to more than one habitat. Data on each habitat were summarized in factsheets containing habitat description, distribution map, corresponding syntaxa and characteristic species combination. Conclusions EUNIS habitats were characterized for the first time in terms of their species composition and distribution, based on a classification of a European database of vegetation plots using the newly developed electronic expert system EUNIS-ESy. The data provided and the expert system have considerable potential for future use in European nature conservation planning, monitoring and assessment. The previous versions of the expert system and related reports were produced within a contract from the European Environment Agency to Wageningen Environmental Research and Masaryk University. The opinions expressed are those of the contractor and do not represent the Agency's official position. EVA data management and preparation of this paper were supported by the Czech Science Foundation (project no. 19-28491X to MC, LT, IK, TP, CM, JDa, MH, PN, DZ, GB, AJ, AKu, ZL and DV). IB and JAC were supported by the Basque Government (project no. T936-16). TB, ET, and LK were supported by the Ministry of Science and Higher Education of the Russian Federation (TB and ET project no. AAAA-A18-118052590019-7; LK project no. AAAA-A19-119012490096-2).

Mutagenesis, 16 (5), ISSN:0267-8357, ISSN:1464-3804

The potent food mutagen and carcinogen 2-amino-3-methylimidazo[4,5- f]quinoline (IQ) and the structurally related heterocyclic aromatic amines 2-aminoimidazo[4,5-f]quinoline (demethyl-IQ) and 2-amino-1-methylimidazo[4,5-f]quinoline (iso-IQ) were assayed for genotoxicity in the wing somatic mutation and recombination test (SMART) as well as in the sex-linked recessive lethal (SLRL) test in Drosophila melanogaster. In addition, 3-methyl-2-nitroimidazo[4,5-f]quinoline (nitro-IQ), 2-nitrofluorene and 1,8-dinitropyrene were also assayed in the wing spot test. IQ was clearly mutagenic in the SLRL test with highest activity in spermatids. Iso-IQ was more active than IQ whereas demethyl-IQ was inactive in this test. The same pattern of results was obtained in the wing SMART: iso-IQ produced greater than 2-fold higher frequencies of spots than IQ and demethyl-IQ was clearly negative. In addition, nitro-IQ exhibited an approximately equal genotoxic activity as IQ. 2-Nitrofluorene and 1,8-dinitropyrene were both inactive in the wing spot test. These data provide good evidence for a correlation of genotoxic effects in germinal and somatic cells, and for the practical advantage of the wing spot test in Drosophila. Moreover, the results show structure-activity relationships among the heterocyclic aromatic amines and nitro compounds similar to those found in Salmonella.

In an era of the availability of very large databases, the problem of efficient methods to analyze such datasets remains. In large scale forest and landscape monitoring projects for instance, appropriate data mining techniques that can summarize the overall status of landscapes are necessary for planning and implementing follow-up management strategies. We consider a vegetation data set consisting of species data from more than 120 mires spread across Switzerland with a total of 20,134 plots on 2658 vascular and non-vascular plant species. Using species indicator values as proxy for site conditions, we propose some simple strategies for data mining involving multidimensional scaling and nonparametric probability density function estimation, both of which are known in the classical statistical literature. We show how commonly known techniques can be adapted in a novel and effective approach for the specific purpose of analyzing plant species occurrence in the plots to identify site conditions that influence species configurations and species diversity, thus providing important information concerning aspects of large scale vegetation structure. Our results indicate high variations among the mires with respect to site conditions that affect species assemblage and species diversity. While species indicator values continue to be popular as well as subject of much debate and research in the ecological community, our experience shows that careful methods of analysis at large landscape scales can reveal some powerful results, which can be taken up as the starting point for the next level of investigation.

In Switzerland, steppe vegetation is restricted to inner alpine dry valleys in the West (Valais) and the East (Grisons). They harbour many specialist and threatened species. In 2011, a long-term program was established to monitor changes in Swiss habitats of national importance. Here, we studied patterns of vascular plant diversity in a subset of 148 steppe plots of 10 m2 from this program. Plots were either grazed or abandoned. On average, we found 23.3 species per plot. The proportion of steppe specialists and of threatened species out of the total species richness was 29.3% and 12.2%, respectively. The total number of species increased with elevation and peaked at intermediate productivity levels. In contrast, the proportion of threatened and steppe specialist species was generally negatively related to higher productivity and wood cover – two factors considered as indicators of habitat degradation. Abandonment vs. pasture had no effects on total species richness. Productivity was positively related to ecological indicator values for moisture, competition and ruderality, as well as the community mean traits of plant height and specific leaf area, but negatively with the indicator value for light. Similarly, wood cover was positively associated with higher ecological indicator values for productivity, moisture and competition, as well as the community mean trait of plant height, but negatively with the indicator value for light and ruderality. The negative response of threatened and steppe specialist vascular plant species to habitat degrading factors suggests their proportional richness to be a better indicator of habitat quality than the total species richness. We conclude that management regimes which guarantee low productivity and low wood cover and avoid intensification by fertilization and irrigation are essential for the conservation of steppe habitats. Beside periodic clearing of woody species, low-intensity grazing might prevent shrub encroachment.

To assess whether short-term changes in mire vegetation can be detected using the phytosociological approach, paired vegetation releves from two surveys of 112 mire sites of Switzerland were assigned to phytosociological alliances through a numerical approach. About 30% of the plots were assigned to different alliances in the two surveys. These transitions were analysed based on species frequencies and interpreted ecologically using Landolt’s indicator values. Transitions between different alliances were more frequently related to the appearance of new species rather than to the disappearance of species. Transitions from and to peat bog communities were frequent. Many plots with fen vegetation were transformed into other wetland types. Fen-grassland increased in abundance, mainly at the cost of small-sedge fens. To re-establish the function of the Swiss mires as peat producers, we recommend to raise the mean summer water table to a maximum depth of 10 cm.

The mapping and monitoring of Swiss mires has so far relied on a classification system based on expert judgement, which was not supported by a quantitative vegetation analysis and which did not include all wetland vegetation types described in the country. Based on a spatially representative sample of 17,608 releves from 112 Swiss mires, we address the following questions: (1) How abundant are wetland vegetation types (phytosociological alliances) in Swiss mires? (2) How are they distributed across the country––is there a regional pattern? (3) How clearly are they separated from each other? (4) How clear and reliable is their ecological interpretation? Using published wetland vegetation releves and lists of diagnostic species for phytosociological units (associations and alliances) established by experts, we developed a numerical method for assigning releves to units through the calculation of similarity indices. We applied this method to our sample of 17,608 releves and estimated the total area covered by each vegetation type in Switzerland. We found that vegetation types not included in previous mapping were either rare in Switzerland (partly due to mire drainage) or poorly distinguished from other vegetation units. In an ordination, the Swiss mire vegetation formed a triangular gradient system with the Sphagnion medii, the Caricion davallianae and the Phragmition australis as extreme types. Phytosociological alliances were clearly separated in a subset of 2,265 releves, which had a strong similarity to one particular association, but poorly separated across all releves, of which many could not be unequivocally assigned to one association. However, ecological gradients were reflected equally well by the vegetation types in either case. Overall, phytosociological alliances distinguished until now proved suitable schemes to describe and interpret vegetation gradients. Nevertheless, we see the urgent need to establish a data base of Swiss wetland releves for a more reliable definition of some vegetation units.

Botanica Helvetica, 119 (2), ISSN:0253-1453, ISSN:1420-9063

Proanthocyanidins (PAs), also known as condensed tannins, are naturally occurring oligomers and polymers of flavan-3-ol monomer units widely found in the leaves, flowers, fruits, seeds, nuts and barks of many plants. Grape seed proanthocyanidins (GSPs) have been used as nutritional supplements, as antioxidants, in preventing atherosclerosis and cardiovascular diseases, and for dislipidemy treatment. The anthracycline antibiotic adriamycin (Doxorubicin, DXR) is a cancer chemotherapeutic agent that interferes with the topoisomerase II enzyme and generates free radicals. In the present study, GSPs (1.680, 3.375, or 6.750 mg/mL) alone were examined for genotoxicity, and combined with DXR (0.125 mg/mL) for antigenotoxicity, using the standard (ST) and high bioactivation (HB) versions of the wing somatic mutation and recombination test in Drosophila melanogaster. The results observed in both crosses were rather similar. GSPs themselves did not show genotoxicity at the doses used. GSPs suppressed the DNA damage induced by DXR in a dose-dependent manner. Comparison of the frequencies of wing spots in the marker-heterozygous (MH) flies and balancer-heterozygous (BH) flies from both crosses, indicated that induced recombination was the major response for the treatments with DXR alone. The co-treatments demonstrated that GSPs have some anti-mutagenic activity; however, anti-recombinagenic activity was the major response.

The wing Somatic Mutation and Recombination Test (SMART) in D. melanogaster was used to study genotoxicity of the medicinal plant Tabebuia impetiginosa. Lapachol (naphthoquinone) and β-lapachone (quinone) are the two main chemical constituents of T. impetiginosa. These compounds have several biological properties. They induce apoptosis by generating oxygen-reactive species, thereby inhibiting topoisomerases (I and II) or inducing other enzymes dependent on NAD(P)H:quinone oxidoreductase 1, thus affecting cell cycle checkpoints. The SMART was used in the standard (ST) version, which has normal levels of cytochrome P450 (CYP) enzymes, to check the direct action of this compound, and in the high bioactivation (HB) version, which has a high constitutive level of CYP enzymes, to check for indirect action in three different T. impetiginosa concentrations (10%, 20% or 40% w/w). It was observed that T. impetiginosa alone did not modify the spontaneous frequencies of mutant spots in either cross. The negative results observed prompted us to study this phytotherapeuticum in association with the reference mutagen doxorubicin (DXR). In co-treated series, T. impetiginosa was toxic in both crosses at higher concentration, whereas in the HB cross, it induced a considerable potentiating effect (from ~24.0 to ~95.0%) on DXR genotoxity. Therefore, further research is needed to determine the possible risks associated with the exposure of living organisms to this complex mixture., Genetics and Molecular Biology, 32 (2), ISSN:1415-4757, ISSN:1678-4685

As conservation of sensitive habitats is a high priority issue in European environmental policy, there is considerable interest in mapping and monitoring specific habitats of high conservation value. In this study, we discuss the potential of the Swiss mire monitoring program to monitor small area habitats in sufficient detail. The monitoring scheme combines nationwide probability sampling and predictive habitat mapping based on a field data sample. Thus, it is designed to identify spatiotemporal changes at the stand level and to derive hard statistics for the sub-national level. For feasibility reasons, the thematic focus is on semi-quantitative mean indicator values derived from vegetation records. These measures provide robust estimates of essential floristic site conditions. Regression models based on CIR aerial photographs are applied to continuously map respective measures across the sample mires. The present study explores the required investment of data for model-based mapping. Exemplary mapping r...

Propolis is a substance produced by honeybees (Apis mellifera L.). Its components are strong antioxidants and free radical scavengers. The aim of this study was to evaluate the protective effects of a water extract of Brazilian green propolis (WEP) combined with the antitumor agent doxorubicin (DXR) on Drosophila melanogaster wing cells through the somatic mutation and recombination test (SMART). Two different crosses were used: The standard (ST) cross and the high bioactivation (HB) cross. The HB cross is characterized by a constitutively enhanced level of cytochrome P450 which leads to an increased sensitivity to a number of promutagens and procarcinogens. Larvae obtained from these two crosses were chronically treated with different concentrations of WEP (12.5,25.0 and 50.0 mg/mL) alone or combined with DXR (0.125 mg/mL). The results obtained with the two different crosses were rather similar. Neither toxicity nor genotoxicity were observed in WEP treated series. Simultaneous treatment with different concentrations of WEP and DXR led to a reduction in the frequency of recombination compared to the treatment with DXR alone. This anti-recombinogenic effect was proportional to the concentrations applied, indicating a dose-response correlation and can be attributed to the powerful scavenger ability of WEP.

A Mandevilla velutina crude extract was investigated using the mouse micronucleus test (MNT) and the Drosophila melanogaster somatic mutation and recombination test (SMART) using standard (ST) and high bioactivation (HB) crosses. The MNT used 10 mg, 20 mg or 40 mg per 100 g of body weight (bw) of extract with and without 0.2 mg per 100 g bw peritoneal cyclophosphamide. There was no genotoxicity in the negative control or extract only groups and, compared to the cyclophosphamide control, there was a significant reduction in micronucleated polychromatic erythrocytes in all the groups given extract plus cyclophosphamide. For SMART larvae were fed 5 or 10 mg mL-1 of extract for seven days with and without 0.89 mg mL-1 of urethane given on day seven. The ST and HB flies showed no significant differences in spots between the negative control and the extract only groups. The number of urethane-induced spots was reduced by the highest concentration of extract for the ST flies and by both concentrations of extract for the HB flies. The results suggest that M. velutina extract is not genotoxic but is antigenotoxic., Genetics and Molecular Biology, 31 (3), ISSN:1415-4757, ISSN:1678-4685

The Drosophila melanogaster somatic mutation and recombination test (SMART) was used to assess the genotoxicity of surface (S) and bottom (B) water and sediment samples collected from Sites 1 and 2 on the Japaratuba River (Sergipe, Brazil), an area impacted by a petrochemical industrial complex that indirectly discharges treated effluent (produced water) into the river. The genotoxicity tests were performed in standard (ST) cross and high bioactivation (HB) cross flies and were conducted on samples taken in March (dry season) and in July (rainy season) of 2003. Mutant spot frequencies found in treatments with unprocessed water and sediment samples from the test sites were compared with the frequencies observed for similar samples taken from a clean reference site (the Jacarecica River in Sergipe, Brazil) and those of negative (ultrapure water) controls. While samples from the Japaratuba River generally produced greater responses than those from the Jacarecica River, positive responses were detected for both the test and reference site samples. All the water samples collected in March 2003 were genotoxic. In July 2003, the positive responses were restricted to water samples collected from Sites 1 B and 2 S in the ST cross. The genotoxicity of the water samples was due to mitotic recombination, and the samples produced similar genotoxic responses in ST and HB flies. The spot frequencies found in the July water samples were considerably lower than those for the March water samples, suggesting a seasonal effect. The only sediment samples that were genotoxic were from Site 1 (March and July) and from the Jacarecica River (March). The genotoxins in these samples produced both somatic mutation (limited to the Site 1 sample in HB flies) and recombination. The results of this study indicate that samples from both the Japaratuba and Jacarecica Rivers were genotoxic, with the most consistently positive responses detected with Site 1 samples, the site closest to the putative pollution source.

Neurocysticercosis, the most common parasitic disease of the central nervous system, is caused by cysticerci of the helminth Taenia solium, which is prevalent in developing countries and is reemerging in affluent societies. This helminth is associated with brain tumors and hematological malignancies in humans. In the present study, we analyzed the genotoxicity of vesicular fluid (VF) and a saline extract (SE) of T. solium metacestodes in the Drosophila melanogaster wing somatic mutation and recombination test (SMART). Third-instar larvae derived from standard and high bioactivation crosses were treated for approximately 48 hr with 12.5, 25.0, and 50.0 μg/ml of VF and SE of T. solium metacestodes. Negative (phosphate buffered saline) and positive (10 mM urethane) controls were also included. The results showed that the two test compounds were genotoxic in both crosses of Drosophila. Nevertheless, further research is needed to determine the genotoxic potential of specific compounds present in VF and SE and their role in the development of cancer. Environ. Mol. Mutagen., 2006. © 2006 Wiley-Liss, Inc.

The detection and evaluation of changes in vegetation patterns is a prerequisite for monitoring programs. The Swiss mire monitoring program aims to assess the changes in mire vegetation in order to examine the efficiency of the management measures. A promising way to explore and detect vegetation structure and vegetation change is the application of predictive vegetation mapping that combines image classification and predictive habitat distribution models. These models deal with predictor variables derived from remotely sensed spectral data and from environmental variables such as a digital surface model (DSM). Low accuracy of environmental data to predict vegetation at the local scale is due to the difficulties to capture dominant fine-scale enironmental gradients. Using high resolution spectral and topographical data sets of 50 cm pixel size and below, the study presented here aims to improve the simulation of local-scale vegetation properties.

Tamoxifen (TAM) is an anti-oestrogen used for treatment and prevention of human breast cancer, but it is also related to human endometrial and uterine cancer. The wing spot test in Drosophila melanogaster was employed to determine the genotoxic effects of TAM and 4-nitroquinoline-1-oxide (4-NQO), a carcinogen that produces adducts similar to TAM-DNA adducts detected in rodent liver and human liver microsomes. As Drosophila spp. have no oestrogen receptor, no effects can result in binding of TAM to a receptor. Chronic treatments with TAM citrate were performed with 3-day-old larvae of the standard (ST) and high bioactivation (HB) crosses of the wing spot test at concentrations of 0.66, 1.66 and 3.33 mM. In addition, the carcinogen 4-NQO was administered at 2.5 and 5.0 mM. Somatic spots on normal wings from marker-heterozygous flies and on serrate wings from balancer-heterozygous flies were scored to determine mutation and recombination events in somatic cells for each compound. The results showed genotoxic effects of TAM at 1.66 and 3.33 mM in the ST cross only and without a clear dose-response effect. This suggests a weak genotoxicity of this anti-oestrogen. The negative results obtained with TAM in the HB cross may indicate efficient detoxification of the compound by the increased xenobiotic metabolism present in this cross. As reported before, 4-NQO showed genotoxic effects in the ST cross with a clear dose-response effect. For the first time, we report enhanced effects of this compound in the HB cross. It is concluded that the genotoxicity of TAM in the Drosophila wing spot test is different from that of 4-NQO.

Stryphnodendron adstringens (Mart.) Coville, a medicinal plant that grows in the "cerrados" (a savanna ecosystem) of Brazil, popularly known as "Barbatimao," is an important source of tannins (polyphenols). In Brazil, it is used in industry (mainly as vegetable tanning) and also in traditional medicine for the treatment of various diseases. In the present study, a phytotherapeutic extract from S. adstringens stem bark was evaluated for mutagenic and recombinagenic effects using the wing spot test of Drosophila melanogaster (somatic mutation and recombination test, SMART), and for chromosome damage in germ cells using the Drosophila sex-chromosome loss test (ring-X loss). For SMART, the standard as well as the high bioactivation fly crosses were used; the latter cross is characterized by a high sensitivity to promutagens and procarcinogens. Third-instar larvae from these two crosses were treated for 48 hr with different concentrations (66%, 75%, and 100%) of the phytotherapeutic extract. The wings of the emerging adults were analyzed for the occurrence of different types of mutant spots. No statistically significant differences in spot frequencies between controls and treated series were observed. For the ring-X loss test, adult males were fed with the same concentrations of the extract as in the wing spot test. No statistically significant increases in ring-X losses were observed. The results of our experiments suggest that the phytotherapeutic extract from S. adstringens stem bark is not genotoxic in somatic and germ cells of D. melanogaster.

The wing spot test in Drosophila melanogaster is a suitable system for the analysis of genotoxic activity of compounds that need metabolic transformation to render them active. We have analysed the genotoxicity of three fungicides for which it was reported that the metabolic processes taking place in vivo may determine their activity. The compounds analysed are captan, maneb, zineb and ethylenethiourea (ETU) (a metabolic derivative of ethylenebisdithiocarbamates like maneb and zineb). We have also evaluated the ability of ETU to form genotoxic derivatives in vivo analysing this compound in combined treatments with sodium nitrite. Both standard and high bioactivation NORR strains have been used. Captan, usually considered a mutagen in vitro but a non-mutagen in vivo, gave negative results in the wing spot test with both crosses. Positive results were obtained for maneb in the standard cross and for ETU in both the standard and the high bioactivation cross. The genotoxicities of maneb and ETU were higher when treatments were made on media in which nitrosation is favoured. A low absorption of the fungicide and an inefficient availability of the compound in the target may explain negative results obtained with zineb in both crosses. The results obtained in this study with the wing spot test demonstrate once again the suitability of this in vivo assay, in which absorption, distribution and metabolism processes take place, for the evaluation of genotoxicity of compounds to which humans are exposed.

Mutagenesis, 17 (2), ISSN:0267-8357, ISSN:1464-3804

The modulating effects of tannic acid (TA) on somatic mutation and mitotic recombination induced by methylmethanesulfonate (MMS), nitrogen mustard (HN2), and mitomycin C (MMC) were evaluated in the standard (ST) cross of the wing spot test in Drosophila melanogaster using co- and posttreatment protocols. It was shown that TA alone did not modify the spontaneous frequencies of single and twin spots, which means that this polyphenol neither acts as a genotoxin nor exerts any antigenotoxic effect over spontaneous DNA lesions. However, the simultaneous administration of genotoxins with TA can lead to considerable alterations of the frequencies of induced wing spots in comparison to those with administration of the genotoxins alone. In fact, TA produced a significant increase in HN2-induced wing spots with enhancements between 90 and 160%. For MMS, the enhancement was 38% in the highest TA concentration tested. In contrast, a significant protective action of this polyphenol was observed in combined treatments with MMC (64 to 99% inhibition). Moreover, the data from TA posttreatments demonstrated that this agent is not effective in exerting protective or enhancing effects on the genotoxicity of MMS, HN2, or MMC. One feasible mechanism of TA action is its interaction with the enzyme systems catalyzing the metabolic detoxification of MMS and HN2, which may also be involved in the bioactivation of MMC.

Among the great variety of genotoxicity assays available, the wing spot test in Drosophila melanogaster has some characteristics that make it very suited for the screening of genotoxic activity, i.e., it is an easy and inexpensive assay using a eukaryotic organism in vivo. One of the most interesting characteristics of the assay is its capacity to detect genotoxic activity of promutagens without the necessity of an exogenous metabolic activation system. In this paper we present results obtained with a recently developed high bioactivation cross of the wing spot test (NORR cross). The positive results obtained with the five well-known procarcinogens 7,12-dimethylbenz[ a ]anthracene, N -nitrosopyrrolidine, p -dimethylaminoazobenzene, diethylnitrosamine and urethane clearly show that the NORR strains are similar to the other high bioactivation strains previously described, but they lack their methodological disadvantages. We have tested six insecticides, which are characterised by having contradictory results in other genotoxicity tests, using both the standard and the high bioactivation (NORR) cross. The six insecticides analysed are the pyrethroid allethrin, the methylenedioxyphenolic compound piperonyl butoxide, the chlorinated hydrocarbons dieldrin and endrin, and the organophosphates dimethoate and malathion. We obtained negative results for all six compounds. Our results show the suitability of the wing spot test for the evaluation of compounds at the first level of genotoxicity testing.

The fruit fly Drosophila melanogaster with its well developed array of genotoxicity test systems has been used in a number of studies on antigenotoxicity of various compounds and mixtures. In recent years, the newly developed Somatic Mutation and Recombination Tests (SMART) have mainly been employed. These one-generation tests make use of the wing or eye imaginal disc cells in larvae and have proven to be very efficient and sensitive. They are based on the principle that the loss of heterozygosity of suitable recessive markers can lead to the formation of mutant clones of cells that are then expressed as spots on the wings or eyes of the adult flies. We have employed the wing spot test with the two markers multiple wing hairs ( mwh , 3-0.3) and flare ( flr , 3–38.8). Three-day-old larvae, trans-heterozygous for these markers, are treated chronically or acutely by oral administration with the test compound(s) or complex mixtures. For antigenotoxicity studies, chronic co-treatments can be used, as well as separate pre-treatments with an antigenotoxic agent followed by a chronic treatment with a genotoxin. After eclosion, the wings of the adult flies are scored for the presence of single and twin spots. These spots can be due to different genotoxic events: either mitotic recombination or mutation (deletion, point mutation, specific types of translocation, etc.). The analysis of two different genotypes (one with structurally normal chromosomes, one with a multiply inverted balancer chromosome) allows for a quantitative determination of the recombinagenic activity of genotoxins. Results of two separate studies are presented: (1) instant coffee has antirecombinagenic but not antimutagenic activity in the wing spot test; and (2) ascorbic acid and catechin are able to protect against in vivo nitrosation products of methyl urea in combination with sodium nitrite.

The in vivo nitrosation capacity of third-instar larvae of Drosophila melanogaster was assessed using the wing somatic mutation and recombination test (SMART). Larvate derived from two different crosses, the standard cross (ST) and the high bioactivation cross (HB) both involving the recessive wing cell markers multiple wing hairs (mwh) and flare (flr3), were used. The HB cross is characterised by an increased cytochrome P450-dependent bioactivation capacity for promutagens and procarcinogens. The larvae were treated either with methyl urea, sodium nitrite or its combination. N-Nitrosomethylurea was used as a positive control. The wings of the resulting flies were analysed for the occurrence of mutant spots produced by various types of mutational events or by mitotic recombination. Methyl urea is negative in the ST and the HB cross, whereas sodium nitrite is weakly genotoxic in both crosses. However, the combination of both compounds produces highly increased frequencies of mutations and recombinations predominantly in the HB cross. The genotoxic effects produced by the combined treatments were considerably increased when mashed potatoes or an agar-yeast medium were used for the treatment instead of the standard instant medium. Treatment of larvae with the mixture resulting from the in vitro reaction of nitrosation precursors also resulted in high frequencies of induced spots comparable to those recorded with the potent genotoxin N-nitrosomethylurea. Further experiments showed that the genotoxic effect resulting from the in vivo exposure to nitrosation precursors can be reduced by co-treatment with catechin, a known nitrosation inhibitor. The present study demonstrates that the wing spot test is well suited for the determination of genotoxicity produced by in vivo nitrosation processes and for the study of their modulation by individual compounds or dietary complex mixtures.

Integerrimine (ITR), a pyrrolizidine alkaloid from Senecio brasiliensis, was tested for genotoxicity using the wing somatic mutation and recombination test (SMART) in Drosophila melanogaster. The compound was administered by chronic feeding (48 hours) of 3-day-old larvae. Two different crosses involving the markers flare (flr) and multiple wing hairs (mwh) were used, that is, the standard (ST) cross and the high bioactivation (HB) cross, which has a high cytochrome P450-dependent bioactivation capacity. In both crosses, the wings of two types of progeny were analyzed, that is, inversion-free marker heterozygotes and balancer heterozygotes carrying multiple inversions. ITR was found to be equally potent in inducing spots in a dose-related manner in the marker heterozygotes of both crosses. This indicates that the bioactivation capacity present in larvae of the ST cross is sufficient to reveal the genotoxic activity of ITR. In the balancer heterozygotes of both crosses, where all recombinational events are eliminated due to the inversions, the frequencies of induced spots were considerably reduced which documents the recombinagenic activity of ITR. Linear regression analysis of the dose response relationships for both genotypes shows that 85% to 90% of the wing spots are due to mitotic recombination.

An increasing number of studies have reported on forest declines and vegetation shifts triggered by drought. In the Swiss Rhone valley (Valais), one of the driest inner-Alpine regions, the species composition in low elevation forests is changing: The sub-boreal Scots pine (Pinus sylvestris L.) dominating the dry forests is showing high mortality rates. Concurrently the sub-Mediterranean pubescent oak (Quercus pubescens Willd.) has locally increased in abundance. However, it remains unclear whether this local change in species composition is part of a larger-scale vegetation shift. To study variability in mortality and regeneration in these dry forests we analysed data from the Swiss national forest inventory (NFI) on a regular grid between 1983 and 2003, and combined it with annual mortality data from a monitoring site. Pine mortality was found to be highest at low elevation (below 1000 m a.s.l.). Annual variation in pine mortality was correlated with a drought index computed for the summer months prior to observed tree death. A generalized linear mixed-effects model indicated for the NFI data increased pine mortality on dryer sites with high stand competition, particularly for small-diameter trees. Pine regeneration was low in comparison to its occurrence in the overstorey, whereas oak regeneration was comparably abundant. Although both species regenerated well at dry sites, pine regeneration was favoured at cooler sites at higher altitude and oak regeneration was more frequent at warmer sites, indicating a higher adaptation potential of oaks under future warming. Our results thus suggest that an extended shift in species composition is actually occurring in the pine forests in the Valais. The main driving factors are found to be climatic variability, particularly drought, and variability in stand structure and topography. Thus, pine forests at low elevations are developing into oak forests with unknown consequences for these ecosystems and their goods and services.

A groups of six chemical compounds was tested in parallel in two different somatic genotoxicity assays in Drosophila melanogaster, the wing somatic mutation and recombination test (SMART) and the white-ivory eye spot test. The wing spot test makes use of the wing cell markers multiple wing hairs (mwh) and flare (flr) and detects both mitotic recombination and various types of mutational events. The white-ivory eye spot test makes use of the white-ivory (wi) quadruplication and detects the somatic reversion of the recessive eye color mutation wi to the wild-type (w+). Three- or two-day-old larvae were fed chronically with the compounds ethylnitrosourea (ENU), N-nitrosopyrrolidine (NNP), caffeine (CAF), chromium (VI) oxide (CRO), potassium chromate (POC), and 2,4-dichlorophenoxyacetic acid (2,4-D). All six compounds are genotoxic to various degrees in the wing spot test. The percentage of the genotoxic activity that is due to mitotic recombination was between 84% and 91% for the hexavalent chromium compounds CRO and POC and about 68% for 2,4-D. In contrast, ENU and NNP showed only 46% and 25% recombinagenic activity, respectively. In the white-ivory eye spot test, the three compounds (CRO, POC, and 2,4-D) with high recombinagenic activity and CAF were clearly nongenotoxic, whereas only ENU and NNP gave a positive response. From these results, it is concluded that the spectrum of genotoxic events detected by the two assays is different. In particular, the white-ivory eye spot test appears not to detect mitotic recombination the way the wing spot test does.

The genotoxicity of three polycyclic aromatic hydrocarbons (PAHs) and of three of their nitro derivatives was evaluated in the wing Somatic Mutation And Recombination Test (SMART) in Drosophila melanogaster. Two crosses were used, i.e. the standard cross (ST) and the improved high bioactivation cross (HB) which is characterised by an increased sensitivity to the genotoxic effects of promutagens and procarcinogens. Larvae trans-heterozygous for the two recessive wing cell markers multiple wing hairs (mwh) and flare (flr3) were fed with the test compounds for 48 h. The wings of the surviving flies were analysed for the occurrence of single and twin spots. Naphthalene, 1-nitronaphthalene and 1,5-dinitronaphthalene proved to be more genotoxic in the HB cross than in the ST cross. Anthracene showed a clear genotoxic activity only in the HB cross whereas it was negative in the ST cross. 9-Nitroanthracene gave inconsistent results in both crosses. Phenanthrene was negative in the ST cross, but weakly positive in the HB cross. These results demonstrate that the genotoxic activity of these PAHs and their nitro derivatives can be detected with the somatic cells of the wing imaginal discs of larvae with high bioactivation capacity.