Trophoblast-like cells sorted from peripheral maternal blood using flow cytometry: A multiparametric study involving transmission electron microscopy and fetal dna amplification

Three monoclonal antibodies (MAbs) against trophoblast (GB17, GB21, and GB25) and flow cytometry were used to sort trophoblast-like cells (TLCs) from peripheral blood of pregnant women. Sorted TLCs were processed for electron microscopy and fetal DNA amplification of the Y-specific sequences from mothers carrying male fetuses. At the ultra-structural level, most of the nucleated cells had the morphology of leucocytes, suggesting maternal contaminants, and we did not find the characteristic features of the free inter-villous trophoblast cells. Nevertheless, polymerase chain reaction (PCR) analysis showed an amplification of Y-specific sequences in two out of three samples of sorted TLCs. These results suggest that besides the maternal leucocytes, sufficient trophoblast nucleated fetal cells can be obtained using cell enrichment by sorting. This sensitive method holds promise for non-invasive prenatal diagnosis of fetal sex and if sufficient Y(positive) nuclei are found, for the diagnosis of selected numerical chromosome abnormalities.     

Disposal of Mine Wastes Containing Pb and Zn Near the Ocean: An Assessment of Associated Environmental Implications in the Arctic

The past two decades have witnessed an increase in the exploration for, and development of, mineral resources at Arctic latitudes. Such effort has resulted in the commencement of production at a number of different localities in close proximity to the ocean. Since proximity to a source of transportation for the conveyance of concentrates to the market is a primary requirement governing the economic decisions for the mine to proceed, it is logical that ore deposits near a coastline are primary targets for development.

Closely tied to coastal mining developments is the need to dispose of tailings and waste rock in the most economical manner. Prior to the advent of heightened public awareness and environmental concern, and the enactment of pollution legislation, disposal practices were largely based upon convenience. Currently, however, proposed disposal and operating practices receive closer scrutiny, with a view to providing adequate protection for aquatic resources and habitat. This paper summarizes some of the features of three Arctic mines, wastes from which enter the marine environment, and compares the disposal practices used at each to measures of environmental change as indicated by metal concentrations in various media.     

Comparative Analysis of Ecophysiological Characteristics of Stephanodiscus hantzschii Grun. in the Periods of Its Bloom in Recreational Water Bodies

In the periods of summer and autumn bloom of the Stephanodiscus hantzschii Crun. in recreational water bodies, studies on the vertical distribution of chlorophyll a, its contents per unit biomass, efficiency in using photosynthetically active radiation (E_PhAR), and assimilative activity of microalgae were performed. The results confirmed the existence of two ecophysiological forms of St. hantzschii and provided evidence that both forms are typically autotrophic and can efficiently use low-intensity PhAR for photosynthesis.     

Tolerance of calluna vulgaris and peatland plant communities to sulphuric acid deposition

Critical loads offer a unique way of evaluating impacts of acid deposition by quantifying environmental sensitivity. The critical loads of acidity for UK peat soils have been based upon an arbitrary reduction in pH of 0.2 units. This chemical shift needs to be better related to adverse effects on sensitive biological receptors. It is known that effective precipitation pH equates closely to soil solution pH, and the latter is directly linkable to biotic effects of pH change. On continuation of a long-term experiment assessing impacts of simulated acid rain on peat microcosms in a realistic outdoor environment, Calluna vulgaris continued to flourish at acid deposition loads well above the existing critical load. Calluna plants were harvested and analysed, and acid deposition treatments to the microcosms continued to allow natural vegetation to regenerate. A diverse mixture of moorland plants and bryophytes established at acidity treatments well above the existing critical load, and only a very high acid load resulted in no natural regeneration. A critical effective rain pH value of 3.6 is suggested as a basis for setting critical loads. At this pH, Calluna grows well, and a healthy diverse vegetation community re-establishes when harvested. It is suggested that the peat critical load should be set at the acid load that, at any specific site, would result in a mean effective precipitation pH of 3.6.     

Methodological Basis of Ecological Safety Standards for the Technogenic Impact of Mineral Resource Exploitation

A new methodological approach to the development of biological and technological safety standards for the impact of underground mining on the natural biota is proposed.     

Biological productivity of bogs in the middle taiga subzone of Western Siberia

New experimental data on biological productivity of plant communities in oligotrophic and mesotrophic bogs of the middle taiga subzone over the past five years are presented. The relationship between net primary production and the stock of live phytomass is estimated. The stock of necromass in oligotrophic bog ecosystems increases from west to east, while the stock of live phytomass and net primary production decrease.     

Isotope tracing of submarine groundwater discharge offshore Ubatuba, Brazil: results of the IAEA–UNESCO SGD project

Results of groundwater and seawater analyses for radioactive (³H, ²²²Rn, ²²³Ra, ²²⁴Ra, ²²⁶Ra, and ²²⁸Ra) and stable (D and ¹⁸O) isotopes are presented together with in situ spatial mapping and time series ²²²Rn measurements in seawater, direct seepage measurements using manual and automated seepage meters, pore water investigations using different tracers and piezometric techniques, and geoelectric surveys probing the coast. This study represents first time that such a new complex arsenal of radioactive and non-radioactive tracer techniques and geophysical methods have been used for simultaneous submarine groundwater discharge (SGD) investigations. Large fluctuations of SGD fluxes were observed at sites situated only a few meters apart (from 0 cm d⁻¹ to 360 cm d⁻¹; the unit represents cm³/cm²/day), as well as during a few hours (from 0 cm d⁻¹ to 110 cm d⁻¹), strongly depending on the tidal fluctuations. The average SGD flux estimated from continuous ²²²Rn measurements is 17 ± 10 cm d⁻¹. Integrated coastal SGD flux estimated for the Ubatuba coast using radium isotopes is about 7 × 10³ m³ d⁻¹ per km of the coast. The isotopic composition (δD and δ¹⁸O) of submarine waters was characterised by significant variability and heavy isotope enrichment, indicating that the contribution of groundwater in submarine waters varied from a small percentage to 20%. However, this contribution with increasing offshore distance became negligible. Automated seepage meters and time series measurements of ²²²Rn activity concentration showed a negative correlation between the SGD rates and tidal stage. This is likely caused by sea level changes as tidal effects induce variations of hydraulic gradients. The geoelectric probing and piezometric measurements contributed to better understanding of the spatial distribution of different water masses present along the coast. The radium isotope data showed scattered distributions with offshore distance, which imply that seawater in a complex coast with many small bays and islands was influenced by local currents and groundwater/seawater mixing. This has also been confirmed by a relatively short residence time of 1–2 weeks for water within 25 km offshore, as obtained by short-lived radium isotopes. The irregular distribution of SGD seen at Ubatuba is a characteristic of fractured rock aquifers, fed by coastal groundwater and recirculated seawater with small admixtures of groundwater, which is of potential environmental concern and has implications on the management of freshwater resources in the region.     

Biological Activity in a Heavily Organohalogen-Contaminated River Sediment (8 pp)

Background, Aims and Scope Sediments of the Spittelwasser creek are highly polluted with organic compounds and heavy metals due to the discharge of untreated waste waters from the industrial region of Bitterfeld-Wolfen, Germany over the course of more than one century. However, relatively few data have been published about the chloroorganic contamination of the sediment. This paper reports on the content of different (chloro)organic compounds with special emphasis on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), and chlorobenzenes. Existing concepts for the remediation of Spittelwasser sediment include the investigation of natural attenuation processes, which largely depend on the presence of an intact microbial food web. In order to gain more insight in terms of biological activity, we analyzed the capacity of sediment microflora to degrade organic matter by measuring the activities of extracellular hydrolytic enzymes involved in the biogeochemical cycling of carbon, nitrogen, phosphorus and sulfur. Furthermore, the detection of physiologically active bacteria in the sediment, particularly of those known for their capability to reductively dehalogenate organochlorine compounds, illustrates the potential for intrinsic bioremediation processes. Methods PCDD/F and chlorobenzenes were analyzed by gas chromatography(GC)/mass spectrometry and GC/flame ionization detection, respectively. The activities of hydrolytic enzymes were determined from freshly sampled sediment layers using 4-methylumbelliferyl (MUF) or 7-amino-4-methylcoumarin-conjugated model compounds and kinetic fluorescence measurements. Physiologically active bacteria from different sediment layers were microscopically visualized by fluorescence in situ hybridization (FISH). Specific bacteria were identified by 16S rRNA gene amplification and sequencing. Results and Discussion The PCDD/F congener profile was dominated by dibenzofurans. In addition, the presence of specific tetra and pentachlorinated dibenzofurans supported the assumption that extensive magnesium production was one possible source for the high contamination. A range of other chloroorganic compounds, including several isomers of chlorobenzenes, hexachlorocyclohexane and 1,1,1-trichloro-2,2-bis (p-chloro-phenyl)ethane (DDT), was present in the sediment. Activities of extracellular hydrolytic enzymes showed a strong decrease in those sediment layers that were characterized by high contents of absorbable organic halogen (AOX), indicating disturbed organic matter decay. Interestingly, an abnormal increase of cellulolytic enzyme activities below the organochlorine-rich layers was observed, possibly caused by residual cellulose from discharges of sulfite pulping wastes. FISH revealed physiologically active bacteria in most sediment layers from the surface down to the depth of about 60 cm, including members of Desulfitobacterium (D.) and Sulfurospirillum. The presence of D. dehalogenans was confirmed by its partial 16S rRNA gene sequence. Conclusions Results of chemical sediment analyses demonstrated high loads of organochlorine compounds, particularly of PCDD/F. Several years after stopping the waste water discharge to Spittelwasser creek, this sediment remains a main source for pollution of the downstream river system by way of the ongoing mobilization of sediment during high floods. As indicated by our enzyme activity measurements, the decomposition potential for organic matter is low in organochlorine-rich sediment layers. In contrast, the comparably higher enzyme activities in less organochlorine-polluted sediment layers as well as the presence of physiologically active bacteria suggest a considerable potential for natural attenuation. Recommendations and Perspectives From our data we strongly recommend to explore the degradative capacity of sediment microorganisms and the limits for in situ activity towards specific sediment pollutants in more detail. This will give a sound basis for the integration of bioremediation approaches into general concepts to reduce the risk that permanently radiates from this highly contaminated sediment. Submission Editor: Dr. Henner Hollert (Henner.Hollert@urz.uniheidelberg.de)