Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus.

Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter-assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Impact of lakeshore development on green frog abundance

Many amphibian species exhibit metapopulation spatial dynamics and temporally are faced with local population extinction and re-colonization. These natural population fluctuations can exhibit stochastic effects when human-caused alteration and fragmentation of habitats occur during sensitive life-cycle events. In this study, we explored the effects of shoreline development on adult green frogs Rana clamitans melanota on lakes (n=24) of northern Wisconsin. We estimated green frog abundance using both auditory and direct observation surveys. The immediate shoreline habitat was mapped and placed into a Geographical Information System (GIS) for analysis. Adult green frog populations were significantly lower on lakes with varying degrees of shoreline house and cottage development than lakes with little or no development. A negative linear relationship existed between shoreline development densities and the number of adult green frogs. However, house and cottage densities alone did not directly explain this reduction. Analysis of variance (ANOVA) identified that the amount of suitable habitat, not development density, significantly affected green frog abundance. Therefore, greater development densities significantly decrease breeding habitat quality, resulting in lower adult frog abundance. These and other findings suggest that lakeshore development regulations are not protecting sensitive amphibian species.     

Numerical taxonomic studies in the generaTriticum L. andPisum L.

Summary Up till now numerical taxonomy has been applied to cultivated plants only to a limited extent though this method seems to be especially suitable for handling the great diversity of these plants. Moreover the theory of numerical taxonomy has not been studied sufficiently.Therefore studies with cultivated plants were begun with the aim to investigate the fitness of different numerical taxonomic techniques for their classification. Materials used: 1) 24 species ofTriticum and additionally 24 species ofAegilops 2) 1225Triticum taxa from Iran (mostlyT. aestivum) 3) 312 taxa ofPisum (mostlyP. sativum).Using 10 coefficients and 7 clustering methods inTriticum andAegilops in most cases a clear separation of both genera resulted as well as a species arrangement withinTriticum according to the genomes.The main aim when dealing with the wheat from Iran was the investigation of the influence of different character sets. At this the coefficient ofGower and the unweighted pair-group method using arithmetic averages (UPGMA) were applied. The separation of the different species is satisfying clear while the representatives of the different varieties ofT. aestivum could not always be found in one cluster. This effect increased when adding further characters sets.InPisum the influence of 11 similarity coefficients to the dendrogram formation has been investigated. With two-state characters the simple matching coefficient and with mainly multistate characters the Canberra metric gave best results (using UPGMA and information measures SUMRAT and SAMRAT).The hitherto existing results show the complicatedness of the evaluation of numerical taxonomic methods in their application to cultivated plants. Therefore generalizations are not yet possible.

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Numerisch-taxonomische Studien in den GattungenTriticum L. undPisum L.

     

Metal Enrichment in Zlatna, a Romanian Copper Smelting Town

A preliminary survey of metal concentrations in soil and vegetable samples was undertaken in the town of Zlatna, in western Romania that is dominated by a large copper smelter. One data set (town survey) consisted of soil samples taken from sites both in the centre of the town (near smelter and school grounds) and the outskirts and included those from roadside verges and wooded areas. The second data set consisted of soil samples taken from vegetable garden plots together with an associated sample of spring onion (Allium fistulosum). The aim of this study was to measure levels of the elements Cu, Zn, Pb, Cd and As in soil samples from Zlatna in general, and to assess their uptake into home grown vegetables and thus into the food chain. Concentrations of the elements of interest in soils and vegetables peaked southwest of the smelter and also in soils near the school in the town centre. Concentrations of elements in soils from the town, including those near the school, had the following ranges: Cu, 41–12,000, geometric mean 863 μg/g; Zn, 89–15,600, geometric mean 920 μg/g; Pb, 32–7860, geometric mean 740 μg/g; Cd, BDL–329.5, geometric mean 3.35 μg/g; As, 15–1440 geometric mean 223 μg/g; Thus, the residents of Zlatna are potentially exposed to levels of these elements that are higher than the recommended guideline values. Mean concentrations of the toxic elements in spring onions were: Cu, 10.2; Zn, 95; Pb, 11; Cd, 0.8; As 2.6 μg/g dry weight.     

Pre-extraction preparation (fresh, frozen, freeze-dried, or acetone powdered) and long-term storage of fruit and vegetable tissues: effects on antioxidant enzyme activity

Activities of the antioxidant enzymes ascorbate peroxidase, catalase, dehydroascorbate reductase, glutathione reductase, guaiacol peroxidase, monodehydroascorbate reductase, and superoxide dismutase were assayed in honeydew (Cucumis melo L.) fruit and spinach (Spinacia oleracea L.) leaves either as fresh, frozen to -80 degrees C, frozen in liquid nitrogen, freeze-dried, or acetone powder, representing the various ways tissues are treated prior to enzyme extraction. Treated tissues were analyzed following treatment or stored for up to 8 weeks at -80 degrees C. Enzyme activities in fruit frozen with or without liquid nitrogen and leaves frozen with or without liquid nitrogen or freeze-dried were equal to those of fresh tissue. Enzyme activities in freeze-dried or acetone-powdered fruit and leaves and in acetone-powdered tissues were significantly higher or lower than those in fresh tissue. Enzyme activities in both tissues frozen with or without liquid nitrogen and stored for 8 weeks at -80 degrees C changed little; those in freeze-dried and acetone-powdered tissues, however, significantly increased/decreased over the same period. Fresh tissue should be used in antioxidant enzyme assays, but if storage is necessary, tissues should be placed directly into a -80 degrees C freezer.     

Effect of climate change and nitrogen deposition on central-European forests: Regional-scale simulation for South Bohemia

The simulation of forest production until 2100 under different environmental scenarios and current management practices was performed using a process-based model BIOME-BGC previously parameterized for the main Central-European tree species: spruce, pine, beech and oak and adapted to include forest management practices. Climatic scenario HadCM3 used in the simulations was taken from the IPCC database created within the 3rd Assessment Report. It was combined with a scenario of CO₂ concentration development and a scenario of N deposition. The control scenario considered no changes of climatic characteristics, CO₂ concentration and N deposition. Simulation experiment was performed for the test region - South Bohemia - using a 1 km × 1 km grid. The actual data on the regional forest cover were aggregated for each grid cell in such a way that each cell represented an even-aged single-dominant species stand or non-forested area, and a standard management scenario depending on the stand age and species was applied to each cell. The effect of environmental variables was estimated as the difference of simulated carbon pools and fluxes in 2050 under environmental changes and under control scenario.The model simulation for the period to 2050 with only climate change under constant CO₂ concentration and N deposition indicated a small decrease of NPP (median values by species reached −0.9 to −1.7% for different species), NBP (−0.3 to −1.7%) and vegetation carbon (−0.3 to −0.7%), whereas soil C slightly increased. Separate increase of N deposition gave small positive effect on carbon pools (0.8-2.9% for wood C and about 0.5% for soil C) and more expressed effect on carbon fluxes (1.8-4.3% for NPP and 1.0-9.7% for NBP). Separate increase of CO₂ concentration lead to 0.6-2.4% increase of wood C pool and 0.1-0.5% increase of soil C. The positive effects of CO₂ concentration and N deposition were more pronounced for coniferous than for deciduous stands.Replacement of 0.5% of coniferous plantations every year by natural broadleaved stands evoked 10.5% of increase of wood carbon pool due to higher wood density of beech and oak compared to spruce and pine, but slightly decreased soil and litter carbon pools.