Franco Baldi

Molecular characterization of an n-alkane-degrading bacterial community and identification of a new species, Acinetobacter venetianus.

Twenty-five bacterial strains isolated from the Venice lagoon and implicated in the degradation of n-alkanes, n-alkanols, n-alkanals and n-alkanoates were characterized in molecular and physiological terms. The isolates were grouped by amplified ribosomal DNA restriction analysis (ARDRA) into seven clusters, corresponding to seven species, six of which were identified on the basis of 16S rDNA sequencing. Genetic variability among strains was shown by random amplified polymorphic DNA (RAPD). Only strains of the new species Acinetobacter venetianus grew with n-alkanes (C10, C14 and C20) and their respective oxidation products as sole carbon sources. Strains of the other three species identified thrived on n-alkane oxidation products (n-alkanols, n-alkanals, n-alkanoates). The other three species were not able to grow on any of the substrates tested. Analysis of plasmid content showed that only A. venetianus strains harboured plasmids. These plasmids contained sequences homologous to the Pseudomonas oleovorans alkBFGH genes.

Mercury and methyl mercury in higher fungi and their relation with the substrata in a cinnabar mining area

One-hundred-ninety-five specimens of higher fungi and their substrata collected in the Hg mining area of M. Amiata and around Siena (Central Italy), were analyzed for their total Hg content. Whereas wood decomposers and many species of mycorrhizal fungi accumulated the metal at a very low rate, some mycorrhizal species and all the humus decomposers may accumulate up to 100 ..mu..g.g/sup -1/ d.w. of Hg and in the least contaminated sites, up to 63 times as much Hg as the substratum. Instead in mineralized areas the concentration factor rarely exceeded 1. The methyl mercury content of 35 species (almost all edible), ranged between 0.01 and 3.7 ..mu..g.g/sup -1/ d.w. The possible significance of these findings and the toxicologic implications for local consumers are considered.

Characterization by high performance liquid chromatography (HPLC) of the solubilization of phosphorus in iron ore by a fungus

SummaryThe value of iron ore is adversely affected by phosphorus in concentrations over 0.03% by weight. The present research concerns the use of metabolic products of aPenicillium-like fungus to leach insoluble phosphates (hydroxyapatite) from ores. Ion chromatography was used to measure metabolism of glucose into acidic fragments. The rate and products of glucose degradation depended on both the chemical composition of the growth medium (buffered or not) and incubation conditions (shaken or quiescent). The principal products were identified as oxalic acid and isomers of propylene dicarboxylic acid, mainly itaconic acid. Continued, slow metabolism of itaconic acid generates more oxalic acid. Aliphatic acids were not detected. Both iron ore phosphate and calcium phosphate were partially solubilized by either the spent broth or aqueous oxalic acid. Solubilization of ore phosphorus was greatly assisted by hydrochloric acid added to the spent broth in small increments. The data suggest biological alternatives to costly leaching procedures that use only mineral acids.

Heavy Metal Coprecipitation with Hydrozincite [Zn5(CO3)2(OH)6] from Mine Waters Caused by Photosynthetic Microorganisms

ABSTRACT An iron-poor stream of nearly neutral pH polluted by mine tailings has been investigated for a natural phenomenon responsible for the polishing of heavy metals in mine wastewaters. A white mineralized mat, which was determined to be hydrozincite [Zn5(CO3)2(OH)6] by X-ray diffraction analysis, was observed in the stream sediments mainly in spring. The precipitate shows a total organic matter residue of 10% dry weight and contains high concentrations of Pb, Cd, Ni, Cu, and other metals. Scanning electron microscopy analysis suggests that hydrozincite is mainly of biological origin. Dormant photosynthetic microorganisms have been retrieved from 1-year-old dry hydrozincite. The autofluorescent microorganisms were imaged by a scanning confocal laser microscope. A photosynthetic filamentous bacterium, classified asScytonema sp. strain ING-1, was found associated with microalga Chlorella sp. strain SA1. This microbial community is responsible for the natural polishing of heavy metals in the water stream by coprecipitation with hydrozincite.

Mercury pollution in marine sediments near a chloralkali plant: Distribution and availability of the metal

Eighty samples of sediments were collected from the continental shelf of the Northern Tyrrhenian Sea. High concentrations of Hg were found in sea-bottom sediments off the chlor-alkali plant of Rosignano Solvay (maximum 4.06 μg g−1 dry wt) and in two zones to the south. Because of the large differences in carbonate content and in grain size, Hg was determined in the finest particles of the sediments (15-2 μm), which were obtained by a sedimentation cylinder; concentrations of Hg were higher in the fine fraction and the enrichment factor (EF) was correlated with carbonate content. A sequential leaching method was applied to eight samples collected off the chloralkali plant in order to determine the availability of Hg bound to the inorganic or organic fractions of the sediment. The results show that this element is not easily available and that the sediment is a good sink. Hg availability is discussed.

Modulation of chromium(VI) toxicity by organic and inorganic sulfur species in yeasts from industrial wastes.

Two chromium(VI) resistant yeast strains (Candida sp. andRhodosporidium sp.) were isolated from industrial wastes. Four different yeasts, three from the Industrial Yeast Collection and one of pharmaceutical origin, were also studied in relation to chromate toxicity and its alleviation by sulfur species. The growth of yeasts from industrial wastes was inhibited by 50% by high concentrations of Cr(VI):Candida sp. by 4mm Cr(VI) andRhodosporidium sp. by 10mm Cr(VI) in Sabouraud Broth medium. The other Cr(VI)-sensitive yeasts were inhibited by 0.1mm Cr(VI). The general mechanism of chromium resistance inCandida sp. andRhodosporidium sp. was due to reduced uptake of chromium, but not to biological reduction from Cr(VI) to Cr(III). In Cr(VI)-sensitive yeasts, chromium was accumulated as much as 10-fold, as inSaccharomyces cerevisiae. Cr(VI) toxicity inCandida sp. was modulated from Cr(VI)-resistance to Cr(VI)-hypersensitivity depending on the addition of methionine, cysteine, sulfate and djenkolic acid. IfCandida sp. was grown in the presence of S-amino acids, especially methionine, it was more resistant than if the sulfur source was sulfate. When sulfate transport was enhanced by addition of djenkolic acid,Candida sp. became hypersensitive.Rhosporidium sp. was always resistant to Cr(VI) because sulfate transport was inefficient and it assimilated sulfur as S-amino acids. Cr(VI)-sensitive yeasts required larger amounts of S-amino acids, especially methionine, to tolerate Cr(VI) toxicity. Cysteine was toxic forC. famata 6016 above 50 μm,

Trace metal assessment in sediment, molluscs and reed leaves in the Bay of Follonica (Italy)

Abstract Marine surficial sediments, intertidal molluscs and reed leaves collected in the Bay of Follonica are slightly contaminated by Pb, Ni, Zn and Fe. The main pollution sources are the waste water and the atmospheric emission of industrial plants. Probably, the weathering of the pyrite deposits along the eastern coast of Elba Island and adsorption and coprecipitation with hydrous Fe-Mn oxides increase the trace metal concentrations in sediments from the Piombino Channel. Mussels and limpets are contaminated by Hg because of the higher background level of the northern Tyrrhenian Sea.

CHEMICAL LEACHING AND SPECIFIC SURFACE AREA MEASUREMENTS OF MARINE SEDIMENTS IN THE EVALUATION OF MERCURY CONTAMINATION NEAR CINNABAR DEPOSITS

This paper investigates mercury contamination in recent marine sediment of the Tyrrhenian continental shelf near the mouths of three rivers draining an area with cinnabar deposits (M. Amiata, Italy). By means of chemical leaching, two fractions of the total mercury were distinguished: a ‘non-leachable’ fraction, consisting of mercury held in relatively stable forms and a ‘leachable’ fraction, composed of forms that are more weakly bound to the sediments. The two Hg fractions are correlated to the organic matter content, the distribution of which is strictly dependent on the sediment surface area. In order to eliminate the effects of grain size variations among the samples, the Hg concentrations were normalized to the unit of surface area (ng/m2). Following this procedure, the ‘non-leachable’ Hg was found to be concentrated mostly in a belt along the coast, while the ‘leachable’ Hg fraction was shown to accumulate largely in the zones with high depositional dynamics near the mouths of the rivers. Hypotheses to account for the discrepancy between the low concentrations of ‘leachable’ Hg in the sediment of the outer continental shelf and the high concentrations in the fauna of the same area, reported in previous papers, are presented.

Biodiversity of prokaryotic communities in sediments of different sub-basins of the Venice lagoon

Microbial community structure and diversity in the wide and shallow Venice lagoon were assessed, prior to construction of mobile dams, at nine stations representative of four different sub-basins previously selected on the basis of international guidelines for sediment quality. The sediments were mainly anoxic and were colonized by microbial communities the species richness of which was quantitatively correlated with total elemental sulfur and acid-volatile sulfide. Automated ribosomal intergenic spacer analysis clustered the stations into three groups. One station for each group was hence analyzed in detail for bacterial and archaeal diversity by screening of 16S rRNA gene clone libraries. The dominance of Gammaproteobacteria clones (84% with a high proportion of Vibrionaceae, indicator of urban pollution) determined significant divergence of the station adjacent to industrial and metropolitan areas. Bacteroidetes were widespread, especially where prairies of aquatic plants are located. The other two analyzed stations were dominated by bacterial taxa implicated in the sulfur cycle: the anoxygenic photosynthetic Chromatiales, sulfate- and sulfur-reducing Desulfobacterales and Desulfuromonadales, and members of the Alpha- and Epsilonproteobacteria.

The role of reduced sulfur species in the coalescence of polysaccharides in the Adriatic Sea

Abstract Massive mucilage events occur in the Northern Adriatic Sea presumably by cytoplasmic excretions from deteriorated diatoms. During three such events in the summer of 1991, 1997 and 1998 the presence of reduced sulfur species (RSS) was determined in samples of macroaggregate using electrochemical methods (in-phase alternating current (AC) and linear sweep voltammetry). The detected levels of sulfur, expressed as equivalent to sulfide concentrations, were about 200 nM. In the same mucilage samples, concentrations of organic matter were determined in the range from 60 to 600 mg/l of total organic carbon (TOC). The physico-chemical properties of organic matter in the macroaggregates correspond to those of polysaccharides of very high molecular mass. Scanning confocal laser microscopy (SCLM) and fluorescent molecular probes for sugars (the lectins concanvaline A (Con-A)) showed the transformation of polysaccharide polymer structure resulting in the formation of very stable filaments and layers after the treatment of mucilage samples with sodium sulfide. Commercial polysaccharides of bacterial and algal origin (xanthan, carrageenans types I and II, dextran-T-500) have been used to simulate macroaggregate formation under laboratory conditions after treatment with sodium sulfide. Raman spectroscopy indicated that for all model polysaccharides used, sulfide interaction occurred, as evidenced by visible change of the O–H stretching region in the vibration spectra of the water molecules. Our data suggest that the aggregated polysaccharides from the Adriatic Sea are: (1) structurally affected by addition of sulfide as was the case for most of the model polysaccharides, and (2) the stabilizing effect of sulfide on the aggregated polysaccharides is due to the formation of sulfur-organic compounds.