Genetic Diversity of Lepilemur mustelinus ruficaudatus, a Nocturnal Lemur of Madagascar

The genetic polymorphism of natural populations of Lepilemur mustelinus ruficaudatus was studied by protein electrophoresis. We sampled blood from 72 individuals from four populations separated by geographic or anthropogenic barriers from southwestern Madagascar. Six out of 22 enzyme loci showed genetic variation with a degree of polymorphism of 0.273. The expected and observed degree of genetic heterozygosity over all loci is similar to that of other primates (H_e = 0.058, H_o = 0.036). The F-statistics revealed that the four subpopulations were similar with respect to gene structure (F_ST = 0.065, p = 0.016), but the genotypic structures within subpopulations were inconsistent with random mating. For the total of the four subpopulations the proportion of heterozygous individuals was significantly smaller than expected under random mating (F_IS = 0.373, F_IT = 0.414, p < 0.01). These results correspond closely to what is expected considering the low migration ability of individuals of L. m ruficaudatus leading to small and rather isolated inbred populations.     

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)     

Reversibility of stemflow-induced soil acidification in Swedish beech forest

Stemwater running down the trunks of beech (Fagus sylvatica L.) has an acidifying effect on soil near the stem. The deposition of acidifying substances may be two to four times higher close to the stem compared to in the stand in general. To study reversibility of soil acidification, 72 stumps of beech trees were chosen from five different year classes of felling (3, 6, 9, 14-15, 25-30), representing the years when stemflow ceased to affect the soil. The H(+) concentration (pH KCl) in the topsoil (0-5 cm) differed between the distances 10-30 cm and 230-250 cm from the stumps, the soil close to the stem being more acid. The longer the time since felling the smaller the differences in H(+) concentration became. This reduction in soil acidity amounted to ca. 50% after 15 years, and only small further changes occurred over the next ten years. This indicates that soil may not recover fully from acidification, or does so at a rather slow rate after the initial 15 years of recovery.     

Effects of Sphagnum moss and urban runoff on bioavailability of lead and zinc from acidic wetlands of the New Jersey pinelands

The effects of Sphagnum and urban runoff on the bioavailability of metals were tested by adding PbCl2 and ZnCl2 to laboratory microcosms constructed of peat substrate with or without live Sphagnum spp. and planted with Acer rubrum L. seedlings or Vaccinium macrocarpon Ait. shoots. We hypothesized that Sphagnum would increase bioavailability through its acidifying action, while urban runoff would decrease availability through increases in alkalinity. Metals were more available to the test plants treated with acidic swampwater than with alkaline runoff. Sphagnum moss caused increases in tissue concentrations in V. macrocarpon, but not in A. rubrum. The latter species took up more metals when grown on substrate from sites receiving runoff, whereas the former took up more metals from substrate from undisturbed sites, despite the lower substrate metal concentrations. Differences in uptake by the two species may reflect differences in their ability to root in the Sphagnum mat. The results demonstrate that Sphagnum spp. exerts strong species-specific effects on the uptake of metals by vascular plants, and that plant species native to acidic wetlands vary widely in their response to metals in the substrate.     

A study on the behaviour of pesticides and their transformation products in the Scheldt estuary using liquid chromatography-electrospray tandem mass spectrometry

Off-line solid-phase extraction (SPE) combined with liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS-MS) was used to study the estuarine behaviour of the polar pesticides, atrazine, chloridazon, diuron and metolachlor, and their transformation products (TPs), hydroxyatrazine (HA), desisopropylatrazine (DIA), desethylatrazine (DEA), 3,4-dichlorophenylmethylurea (DPMU) and monuron. The compounds were identified by comparing their LC retention times and product-ion spectra with those of standard solutions. In all but one case the detection limits of the method were sufficient to determine the compounds of interest over the entire salinity range in the estuary. The concentrations of the dissolved pesticides ranged from 70 ng l-1 for chloridazon to 1350 ng l-1 for diuron. The levels of TPs were 3-8% of the levels of their parent pesticide. The mixing plots of polar pesticides and their TPs indicated that TPs, which are present in fresh river water, are conservatively transported to the sea and that no additional amounts of TPs are formed during their transport through the estuary. The one exception was HA, of which approximately 10% of the amount transported to the North Sea is formed in the lower part of the estuary by photochemical oxidation of atrazine. The latter was concluded from the ratios of each analyte over the sum total of the parent pesticide and all TPs along the salinity gradient, which proved to be a useful tool for identifying such estuarine transformations.     

Fly-ash-induced oxidative stress and tolerance in Prosopis juliflora L. grown on different amended substrates

Field experiments were conducted to study the impact of metal accumulation on malondialdehyde (MDA), cysteine and non-protein thiol (NPSH) contents in the plants of Prosopis juliflora grown on the fly ash (FA) amended with soil, blue green algae (BGA) biofertilizer, farm yard manure, press mud and Rhizobium inoculation. The analysis of data revealed that the level of MDA, cysteine and NPSH was higher in the roots of the plant than leaves, which was found positively correlated with metal accumulation. An increase of 361.14, 64.25 and 305.62% in MDA, cysteine and NPSH contents, respectively was observed after 45 days in the roots of the plants grown in 100% FA as compared to 100% garden soil (GS). The level of MDA, cysteine and NPSH was found less in the plants grown on various amendments of FA showing ameliorating effect on the toxicity induced due to the accumulation of metals. The decrease in MDA, cysteine and NPSH contents was higher in Rhizobium-inoculated plants as compared to uninoculated plants grown on 100% FA. The results showed a high tolerance potential of the plant, which is further increased by inoculating the plant with FA-tolerant Rhizobium showing feasibility of using P. juliflora in environmental monitoring of FA landfills.