Vanadium-basidiomycete fungi interaction and its impact on vanadium biogeochemistry.

Fungi are well known to strongly interact with metals, thereby influencing metal biogeochemistry in the terrestrial environment. To assess and quantify potential fungi-vanadium (V) interactions, Amanita muscaria, Armillaria cepistipes, Xerocomus badius and Bjerkandera adusta were cultured in media containing soluble V (VOSO4 or NaVO3) or solid-phase V of different chemical forms and oxidation state (V2O3, VO2, V2O5, or V-Ti magnetite slag). All fungi underwent physiological and structural changes, as revealed by alterations in FT-IR peak positions and intensities relative to the control, and morphological changes of mycelia, as observed by scanning electron microscopy. The diametric growth size generally decreased with decreasing oxidation state of V and with increasing concentrations of VOSO4 and NaVO3, implying that V toxicity is dependent on V speciation. The tolerance index, the ratio of treated and control mycelium (dry weight), shows different tendencies, suggesting additional factors influencing fungi weight, such as the formation of extrahyphal crystals. Vanadium accumulation from VOSO4 and NaVO3 medium in all fungi (up to 51.3 mg g-1) shows the potential of fungi to immobilise soluble V, thereby reducing its impacts on environmental and human health. Uptake and accumulation of V in slag was insignificant, reflecting the association of slag V with insoluble crystalline materials. The fungal accumulation of V in medium amended with V-oxides demonstrates the ability of fungi to solubilise solid-phase V compounds, thereby introducing previously immobile V into the V biogeochemical cycle and into the food chain where it may impact ecological and human health. A.muscaria lowered the pH of the medium substantially during cultivation, indicating acidolysis and complexolysis via excretion of organic acids (e.g. oxalic acid). Oxidation of VOSO4 was observed by a colour change of the medium to yellow during B. adusta cultivation, revealing the role of fungally-mediated redox transformation in V (im)mobilisation. The calculated removal efficiencies of soluble V were 40-90% for A. cepistipes and X. badius, but a much lower recovery (0-20%) was observed from V oxides and slag (0-20%) by all fungi. This suggests the probable application of fungi for bio-remediation of mobile/soluble V in contaminated soils but not of V incorporated in the lattice of soil minerals.

An optimised sequential extraction scheme for the evaluation of vanadium mobility in soils.

Reviewing the current state of knowledge about sequential extraction applied for soil vanadium (V) fractionation, we identified an urgent requirement of an sequential extraction (SE) specified for V. Namely, almost all previous SE extracted only 8.4%-48% of total V in soils (excluding residue). Thus, we proposed an eight-step SE for V fractionation in soils according to the knowledge gained from literature and our own dissolution experiments with model minerals. After extracting the mobilisable and adsorbed V with de-ionised water and 5mmol/L phosphate, 1mol/L pyrophosphate was applied to gather organic matter bound V which minimised the artefact dissolving Al and Fe (hydr)oxides occurred when using HNO3-H2O2 for extraction. Extraction with 0.4mol/L NH2OH⋅HCl was highly selective toward manganese oxides. Fractionation of different crystalline Al and Fe (hydr)oxides associated V with 1mol/L HCl, 0.2mol/L oxalate buffer and 4mol/L HCl at 95°C especially improved the extractability of V incorporated with crystalline phase associated V. The suitability of our new SE scheme was confirmed by its higher selectivity against the target phases and higher extraction efficiencies (55%-77% of total V) with model minerals and 6 soils of different properties than previous SE.

In situ chlorophyll fluorescence kinetics as a tool to quantify effects on photosynthesis in Euphorbia cyparissias by a parasitic infection of the rust fungus Uromyces pisi.

BACKGROUND: Photosynthesis is the key process for plant growth and development. The determination of chlorophyll fluorescence kinetics allows the quantification of effects on photosynthetic processes triggered by environmental stress factors such as, e.g., the infection by fungal phytopathogens. The technique is non-invasive, rapid and well suited for experimental field work. RESULTS: Healthy and Uromyces-infected plants of Euphorbia cyparissias were monitored directly in situ in the field using rapid fluorescence kinetics. Non-infected healthy plants show a typical maximum value for the relative variable fluorescence Fv/Fm of around 0.8 with occasional variation between the leaves from the plant top towards the base, while infected plants exhibited a strong gradient to low values at the base. The photosynthetic performance index (PI) showed a higher heterogeneity within the leaves in both plant types. CONCLUSIONS: The non-invasive and rapid measurement of chlorophyll fluorescence induction allows characterizing the photosynthetic capacity of healthy and infected plants and of parts of them directly in the field. The PI, is highly sensitive not only concerning infection, but also towards other local environmental influences.

Complete Genome Sequence of the Cyanogenic Phosphate-Solubilizing Pseudomonas sp. Strain CCOS 191, a Close Relative of Pseudomonas mosselii.

We sequenced the complete genome of the isolate Pseudomonas sp. CCOS 191. This strain is able to dissolve phosphate minerals and form cyanide. The genome sequence is used to establish the phylogenetic relationship of this species.

Uptake and distribution of stable strontium in 26 cultivars of three crop species: oats, wheat, and barley for their potential use in phytoremediation.

The main objective of this study was to investigate the accumulation and distribution of strontium (Sr) in 26 cultivars of wheat (Triticum aestivum L.), husk oat (Avena sativa L) and naked oat (Avena nuda), and barley (Hordeum vulgare L.) for their potential use in phytoremediation.Sr levels had no effect on the accumulation of shoot biomass at tillering or at maturity. Mean shoot Sr concentration of naked oat and barley at tillering was significantly (P<0.05) higher than that of wheat; Neimengkeyimai-1, a naked oat cultivar, had the highest Sr concentrations. At maturity, of four naked oat cultivars, Neimengkeyimai-1 had the highest Sr content at all measured Sr levels. Leaves had the highest Sr concentrations, followed by roots and straw, and then grain with the lowest. Mean enrichment coefficients from soil to shoots ranged from 0.521 to 1.343; the percentage of stable Sr removed from the soil to the shoots at harvest time was more than 1.4% after 120 days. Neimengkeyimai-1 could be used as a model for further research to find more effective cultivars; and naked oat plants could be selected for phytoremediation to clean up contaminated soil.

Early diagenetic processes generate iron and manganese oxide layers in the sediments of Lake Baikal, Siberia.

Distinct layers of iron(III) and manganese(IV) (Fe/Mn) oxides are found buried within the reducing part of the sediments in Lake Baikal and cause considerable complexity and steep vertical gradients with respect to the redox sequence. For the on-site investigation of the responsible biogeochemical processes, we applied filter tube samplers for the extraction of sediment porewater combined with a portable capillary electrophoresis instrument for the analyses of inorganic cations and anions. On the basis of the new results, the sequence of diagenetic processes leading to the formation, transformation, and dissolution of the Fe/Mn layers was investigated. With two exemplary cores we demonstrate that the dissolution of particulate Fe and Mn is coupled to the anaerobic oxidation of CH4 (AOM) either via the reduction of sulphate (SO4(2-)) and the subsequent generation of Fe(II) by S(-II) oxidation, or directly coupled to Fe reduction. Dissolved Fe(II) diffuses upwards to reduce particulate Mn(IV) thus forming a sharp mineral boundary. An alternative dissolution pathway is indicated by the occurrence of anaerobic nitrification of NH4(+) observed at locations with Mn(IV). Furthermore, the reasons and consequences of the non-steady-state sediment pattern and the resulting redox discontinuities are discussed and a suggestion for the burial of active Fe/Mn layers is presented.

Distribution and identification of culturable airborne microorganisms in a Swiss milk processing facility.

Airborne communities (mainly bacteria) were sampled and characterized (concentration levels and diversity) at 1 outdoor and 6 indoor sites within a Swiss dairy production facility. Air samples were collected on 2 sampling dates in different seasons, one in February and one in July 2012 using impaction bioaerosol samplers. After cultivation, isolates were identified by mass spectrometry (matrix-assisted laser desorption/ionization-time-of-flight) and molecular (sequencing of 16S rRNA and rpoB genes) methods. In general, total airborne particle loads and total bacterial counts were higher in winter than in summer, but remained constant within each indoor sampling site at both sampling times (February and July). Bacterial numbers were generally very low (<100 cfu/m(3) of air) during the different steps of milk powder production. Elevated bacterial concentrations (with mean values of 391 ± 142 and 179 ± 33 cfu/m(3) of air during winter and summer sampling, respectively; n=15) occurred mainly in the "logistics area," where products in closed tins are packed in secondary packaging material and prepared for shipping. However, total bacterial counts at the outdoor site varied, with a 5- to 6-fold higher concentration observed in winter compared with summer. Twenty-five gram-positive and gram-negative genera were identified as part of the airborne microflora, with Bacillus and Staphylococcus being the most frequent genera identified. Overall, the culturable microflora community showed a composition typical and representative for the specific location. Bacterial counts were highly correlated with total airborne particles in the size range 1 to 5 µm, indicating that a simple surveillance system based upon counting of airborne particles could be implemented. The data generated in this study could be used to evaluate the effectiveness of the dairy plant's sanitation program and to identify potential sources of airborne contamination, resulting in increased food safety.

Sediment porewater extraction and analysis combining filter tube samplers and capillary electrophoresis.

Careful extraction and analysis of porewater from sediment cores are critical for the investigation of small-scale biogeochemical processes. Firstly, small sample volumes and high spatial resolution are required. Secondly, several chemical species in the anaerobic porewater are sensitive to oxidation when brought in contact with ambient air. Here we present the combination of a special sampling technique and an analytical method for the porewater extraction of a varved sediment core from Lake Baldegg in central Switzerland, using MicroRhizon samplers and a portable capillary electrophoresis (CE) instrument. MicroRhizon filter tubes of 1 mm diameter and 20 mm length are suitable for fast retrieval of particle-free porewater samples directly from the sediment core. Since the time-span between sampling and analysis is less than 20 seconds, oxygen-sensitive Fe(ii) can be analyzed in one go together with Na(+), K(+), Ca(2+), Mg(2+), NH4(+), and Mn(ii) without splitting, acidification or dilution of the sample. The major inorganic cations and anions of the sediment porewater can be determined in less than 15 minutes. Detection limits are in the sub-micromolar concentration range. The capillary electrophoresis instrument used in this study requires sample volumes of only 20 µL. These remarkable small sample volumes allow the minimization of disturbance of the sediment cores and a high spatial resolution of the sediment profile, even in sediments with low water content. The equipment is inexpensive, easy to handle, fully portable and therefore suitable for environmental on-site applications.

Chemistry and bioactivity of an artificial adenosylpeptide B(12) cofactor.

Artificial influence: We describe a semi-artificial adenosylpeptide B(12) that behaves as a cofactor in B(12)-dependent enzymatic reactions and demonstrate that the peptide backbone influences its chemical properties and modulates its bioactivity in vitro and in vivo. Inhibition of the growth of L. delbrueckii is demonstrated, thus providing a potentially powerful approach for the development of antibacterial and antiproliferative compounds.

Coordination chemistry and biological activity of 5'-OH modified quinoline-B12 derivatives.

The consequences of structural modifications at the 5'-OH ribofuranotide moiety of quinoline modified B12 derivatives are discussed in regard of the coordination chemistry, the electrochemical properties and the biological behaviour of the compound.

Detection and differentiation of bacterial spores in a mineral matrix by Fourier transform infrared spectroscopy (FTIR) and chemometrical data treatment.

BACKGROUND: Fourier transform infrared spectroscopy (FTIR) has been used as analytical tool in chemistry for many years. In addition, FTIR can also be applied as a rapid and non-invasive method to detect and identify microorganisms. The specific and fingerprint-like spectra allow - under optimal conditions - discrimination down to the species level. The aim of this study was to develop a fast and reproducible non-molecular method to differentiate pure samples of Bacillus spores originating from different species as well as to identify spores in a simple matrix, such as the clay mineral, bentonite. RESULTS: We investigated spores from pure cultures of seven different Bacillus species by FTIR in reflection or transmission mode followed by chemometrical data treatment. All species investigated (B. atrophaeus, B. brevis, B. circulans, B. lentus, B. megaterium, B. subtilis, B. thuringiensis) are typical aerobic soil-borne spore formers. Additionally, a solid matrix (bentonite) and mixtures of benonite with spores of B. megaterium at various wt/wt ratios were included in the study. Both hierarchical cluster analysis and principal component analysis of the spectra along with multidimensional scaling allowed the discrimination of different species and spore-matrix-mixtures. CONCLUSIONS: Our results show that FTIR spectroscopy is a fast method for species-level discrimination of Bacillus spores. Spores were still detectable in the presence of the clay mineral bentonite. Even a tenfold excess of bentonite (corresponding to 2.1 × 1010 colony forming units per gram of mineral matrix) still resulted in an unambiguous identification of B. megaterium spores.

Detection of bacterial endospores in soil by terbium fluorescence.

Spore formation is a survival mechanism of microorganisms when facing unfavorable environmental conditions resulting in "dormant" states. We investigated the occurrence of bacterial endospores in soils from various locations including grasslands (pasture, meadow), allotment gardens, and forests, as well as fluvial sediments. Bacterial spores are characterized by their high content of dipicolinic acid (DPA). In the presence of terbium, DPA forms a complex showing a distinctive photoluminescence spectrum. DPA was released from soil by microwaving or autoclaving. The addition of aluminium chloride reduced signal quenching by interfering compounds such as phosphate. The highest spore content (up to 10(9) spores per gram of dry soil) was found in grassland soils. Spore content is related to soil type, to soil depth, and to soil carbon-to-nitrogen ratio. Our study might provide a basis for the detection of "hot spots" of bacterial spores in soil.

Limnological study on a newly built drinking water reservoir near Tirana, Albania.

Bovilla Lake is a reservoir constructed 12 years ago for supplying the city of Tirana (Albania) mainly with drinking water. It has a surface area of 4.6 km(2), a maximum depth of originally 60 m and is monomictic with a stratification period from early spring to end of October. The lake is oligotrophic with low nutrient concentrations (e.g. SRP in spring about 8 µg L( -1)) and minor oxygen depletion in the hypolimnion during thermal stagnation. The lake is highly turbid due to severe particle import by several rivers during rain periods. This led to a massive deposition of sediments, lifting the maximum depth to 45 m in 2008. Furthermore, the photic zone reached hardly more than 10 m. Algal species diversity is high; however, diatoms from the genus Cyclotella dominate most of the year both in numbers and biomass. Our study describes for the first time the hydrography and limnology of the Bovilla Reservoir.

Weathering-associated bacteria from the Damma glacier forefield: physiological capabilities and impact on granite dissolution.

Several bacterial strains isolated from granitic rock material in front of the Damma glacier (Central Swiss Alps) were shown (i) to grow in the presence of granite powder and a glucose-NH(4)Cl minimal medium without additional macro- or micronutrients and (ii) to produce weathering-associated agents. In particular, four bacterial isolates (one isolate each of Arthrobacter sp., Janthinobacterium sp., Leifsonia sp., and Polaromonas sp.) were weathering associated. In comparison to what was observed in abiotic experiments, the presence of these strains caused a significant increase of granite dissolution (as measured by the release of Fe, Ca, K, Mg, and Mn). These most promising weathering-associated bacterial species exhibited four main features rendering them more efficient in mineral dissolution than the other investigated isolates: (i) a major part of their bacterial cells was attached to the granite surfaces and not suspended in solution, (ii) they secreted the largest amounts of oxalic acid, (iii) they lowered the pH of the solution, and (iv) they formed significant amounts of HCN. As far as we know, this is the first report showing that the combined action of oxalic acid and HCN appears to be associated with enhanced elemental release from granite, in particular of Fe. This suggests that extensive microbial colonization of the granite surfaces could play a crucial role in the initial soil formation in previously glaciated mountain areas.

Effect of iron and phosphate on bacterial cyanide formation determined by methemoglobin in two-dimensional gradient microcultivations.

Cyanide formation by microorganisms is typically observed during early stationary growth phase and is facilitated by the presence of iron(III) and inorganic phosphate. Extracellular free cyanide in aqueous solutions might readily react with methemoglobin added and can be determined by UV/VIS spectroscopy. As alternative to existing methods, this provided the basis for an analytical method which has not been used previously for the determination of cyanide in bacterial cultivations. We successfully applied the technique to study the combined effect of both iron(III) and phosphate on the cyanide formation by Pseudomonas fluorescens, which we used as model organism known for its ability to form HCN. Information on the combined effect of iron and phosphate was obtained by using commercially available 24-well microtiter plates as two-dimensional gradient systems. After its reaction with methemoglobin, cyanide was measured reproducibly at the wavelength of 427 nm. We found a combined effect of both iron and phosphate. In the absence of inorganic phosphate, cyanide formation was stimulated considerably by increasing concentrations of iron(III), although the effect on the bacterial growth of P. fluorescens was almost insignificant. This suggests that iron is more important than inorganic phosphate for the cyanogenesis by P. fluorescens.

Host-plant selectivity of rhizobacteria in a crop/weed model system.

Belowground microorganisms are known to influence plants' performance by altering the soil environment. Plant pathogens such as cyanide-producing strains of the rhizobacterium Pseudomonas may show strong host-plant selectivity. We analyzed interactions between different host plants and Pseudomonas strains and tested if these can be linked to the cyanide sensitivity of host plants, the cyanide production of bacterial strains or the plant identity from which strains had been isolated. Eight strains (four cyanide producing) were isolated from roots of four weed species and then re-inoculated on the four weed and two additional crop species. Bacterial strain composition varied strongly among the four weed species. Although all six plant species showed different reductions in root growth when cyanide was artificially applied to seedlings, they were generally not negatively affected by inoculation with cyanide-producing bacterial strains. We found a highly significant plant species x bacterial strain interaction. Partitioning this interaction into contrasts showed that it was entirely due to a strongly negative effect of a bacterial strain (Pseudomonas kilonensis/brassicacearum, isolated from Galium mollugo) on Echinochloa crus-galli. This exotic weed may not have become adapted to the bacterial strain isolated from a native weed. Our findings suggest that host-specific rhizobacteria hold some promise as biological weed-control agents.

Influence of plant diversity and elevated atmospheric carbon dioxide levels on belowground bacterial diversity.

BACKGROUND: Changes in aboveground plant species diversity as well as variations of environmental conditions such as exposure of ecosystems to elevated concentrations of atmospheric carbon dioxide may lead to changes in metabolic activity, composition and diversity of belowground microbial communities, both bacterial and fungal. RESULTS: We examined soil samples taken from a biodiversity x CO2 grassland experiment where replicate plots harboring 5, 12, or 31 different plant species had been exposed to ambient or elevated (600 ppm) levels of carbon dioxide for 5 years. Analysis of soil bacterial communities in these plots by temporal temperature gradient gel electrophoresis (TTGE) showed that dominant soil bacterial populations varied only very little between different experimental treatments. These populations seem to be ubiquitous. Likewise, screening of samples on a high-resolution level by terminal restriction fragment length polymorphism (T-RFLP) showed that increased levels of carbon dioxide had no significant influence on both soil bacterial community composition (appearance and frequency of operational taxonomic units, OTUs) and on bacterial richness (total number of different OTUs). In contrast, differences in plant diversity levels had a significant effect on bacterial composition but no influence on bacterial richness. Regarding species level, several bacterial species were found only in specific plots and were related to elevated carbon dioxide or varying plant diversity levels. For example, analysis of T-RFLP showed that the occurrence of Salmonella typhimurium was significantly increased in plots exposed to elevated CO2 (P < 0.05). CONCLUSION: Plant diversity levels are affecting bacterial composition (bacterial types and their frequency of occurrence). Elevated carbon dioxide does not lead to quantitative alteration (bacterial richness), whereas plant diversity is responsible for qualitative changes (bacterial diversity).

Formation of water-soluble metal cyanide complexes from solid minerals by Pseudomonas plecoglossicida.

A few Pseudomonas species are able to form hydrocyanic acid (HCN), particularly when grown under glycine-rich conditions. In the presence of metals, cyanide can form water-soluble metal complexes of high chemical stability. We studied the possibility to mobilize metals as cyanide complexes from solid minerals using HCN-forming microorganisms. Pseudomonas plecoglossicida was cultivated in the presence of copper- and nickel-containing solid minerals. On powdered elemental nickel, fast HCN generation within the first 12 h of incubation was observed and water-soluble tetracyanaonickelate was formed. Cuprite, tenorite, chrysocolla, malachite, bornite, turquoise, millerite, pentlandite as well as shredded electronic scrap was also subjected to a biological treatment. Maximum concentrations of cyanide-complexed copper corresponded to a solubilization of 42% and 27% when P. plecoglossicida was grown in the presence of cuprite or tenorite, respectively. Crystal system, metal oxidation state and mineral hydrophobicity might have a significant influence on metal mobilization. However, it was not possible to allocate metal mobilization to a single mineral property. Cyanide-complexed gold was detected during growth on manually cut circuit boards. Maximum dicyanoaurate concentration corresponded to a 68.5% dissolution of the total gold added. These findings represent a novel type of microbial mobilization of nickel and copper from solid minerals based on the ability of certain microbes to form HCN.

Bioaerosol formation during grape stemming and crushing.

Indoor formation of airborne particles during pre-fermentation grape processing was assessed by particle counting using laser particle sizers. Particle numbers of four different aerodynamic size classes (0.3 to 0.5 microm, 0.5 to 1 microm, 1 to 5 microm, and >5 microm) were determined during unloading of harvest containers and subsequent grape stemming and crushing. Regarding these size classes, composition before grape handling was determined as 87.9%, 10.4%, 1.7%, and 0.1%, respectively, whereas the composition changed during grape handling to 50.4%, 15.2%, 33.0%, and 1.5%, respectively. Airborne bacteria and fungi originating from grape processing were collected by impactor and liquid impinger samplers. Grape handling resulted in a sixfold increase in total (biological and non-biological) airborne particles. The generation of bacterial and fungal aerosols was associated mostly with particles of aerodynamic diameters>5 microm (mainly 7 to 11 microm) as determined by flow cytometry. This fraction was increased 150fold in relation to background levels before grape crushing. Maximum concentrations of culturable bacteria reached 485,000 colony forming units (cfu/m3), whereas 146,000 cfu of fungi and yeasts were detected per cubic meter of air. Culturable Gram-negative bacteria occurred only in small numbers (180 cfu/m3). In relation to the total number of airborne particles emitted, culturable microorganisms comprised 0.1% to 0.2%. As soon as grape crushing was stopped, particle concentrations decreased rapidly either due to passive settling or due to air currents in the occupational indoor environment reaching background levels.

The quality of Albanian natural waters and the human impact.

Albania possesses a wealth of aquatic ecosystems, many of enormous natural and biological value, such as the Lakes Ohrid, Prespa and Shkodra, glacial lakes, river valleys, and coastal lagoons. Although many habitats are highly polluted by inorganic and organic wastes, detailed knowledge on the water quality is still lacking. For the first time, an environmental assessment of the water quality is presented and the main polluting sources identified. As a consequence, a systematic control and the establishment of routine monitoring of surface and groundwater is proposed, which elucidates the present environmental state and helps to develop new strategies of waste and wastewater management. It would help allow Albania to reach an international standard in environmental protection, as a part of UNECE Convention, the Mediterranean Action Plan, the MAP/UNEP Medpol Program and the Basel Convention.