Vincenza Andreoni

Bioremediation and monitoring of aromatic-polluted habitats

Bioremediation may restore contaminated soils through the broad biodegradative capabilities evolved by microorganisms towards undesirable organic compounds. Understanding bioremediation and its effectiveness is rapidly advancing, bringing available molecular approaches for examining the presence and expression of the key genes involved in microbial processes. These methods are continuously improving and require further development and validation of primer- and probe-based analyses and expansion of databases for alternative microbial markers. Phylogenetic marker approaches provide tools to determine which organisms are present or generally active in a community; functional gene markers provide only information concerning the distribution or transcript levels (deoxyribonucleic acid [DNA]- or messenger ribonucleic acid [mRNA]-based approaches) of specific gene populations across environmental gradients. Stable isotope probing methods offer great potential to identify microorganisms that metabolize and assimilate specific substrates in environmental samples, incorporating usually a rare isotope (i.e., 13C) into their DNA and RNA. DNA and RNA in situ characterization allows the determination of the species actually involved in the processes being measured. DNA microarrays may analyze the expression of thousands of genes in a soil simultaneously. A global analysis of which genes are being expressed under various conditions in contaminated soils will reveal the metabolic status of microorganisms and indicate environmental modifications accelerating bioremediation.

Intrinsic bioremediability of an aromatic hydrocarbon-polluted groundwater: diversity of bacterial population and toluene monoxygenase genes.

The functional and phylogenetic biodiversity of bacterial communities in a benzene, toluene, ethylbenzene and xylene (BTEX)-polluted groundwater was analysed. To evaluate the feasibility of using an air sparging treatment to enhance bacterial degradative capabilities, the presence of degrading microorganisms was monitored. The amplification of gene fragments corresponding to toluene monooxygenase (tmo), catechol 1,2-dioxygenase, catechol 2,3-dioxygenase and toluene dioxygenase genes in DNA extracted directly from the groundwater samples was associated with the presence of indigenous degrading bacteria. Five months of air injection reduced species diversity in the cultivable community (as calculated by the Shannon-Weaver index), while little change was noted in the degree of biodiversity in the total bacterial community, as characterised by denaturing gradient gel electrophoresis (DGGE) analysis. BTEX-degrading strains belonged to the genera Pseudomonas, Microbacterium, Azoarcus, Mycobacterium and Bradyrhizobium. The degrading capacities of three strains in batch liquid cultures were also studied. In some of these microorganisms different pathways for toluene degradation seemed to operate simultaneously. Pseudomonas strains of the P24 operational taxonomic unit, able to grow only on catechol and not on BTEX, were the most abundant, and were present in the groundwater community at all stages of treatment, as evidenced both by cultivation approaches and by DGGE profiles. The presence of different tmo-like genes in phylogenetically distant strains of Pseudomonas, Mycobacterium and Bradyrhizobium suggested recent horizontal gene transfer in the groundwater.

Arsenic-resistant bacteria associated with roots of the wild Cirsium arvense (L.) plant from an arsenic polluted soil, and screening of potential plant growth-promoting characteristics

A rhizobacterial community, associated with the roots of wild thistle Cirsium arvense (L.) growing in an arsenic polluted soil, was studied by fluorescence in situ hybridization (FISH) analysis in conjunction with cultivation-based methods. In the bulk, rhizosphere, and rhizoplane fractions of the soil, the qualitative picture obtained by FISH analysis of the main phylogenetic bacterial groups was similar and was predominantly comprised of Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria. The arsenic-resistant isolates belonged to 13 genera, the most abundant being those of Bacillus, Achromobacter, Brevundimonas, Microbacterium, and Ochrobactrum. Most bacteria grew in the presence of high arsenic concentrations (over 100mM arsenate and 10mM arsenite). Most strains possessed the ArsC, ArsB and ACR3 genes homologous to arsenate reductase and to the two classes of arsenite efflux pumps, respectively, peculiar to the ars operon of the arsenic detoxification system. ArsB and ACR3 were present simultaneously in highly resistant strains. An inconsistency between 16S rRNA phylogenetic affiliations and the arsenate reductase sequences of the strains was observed, indicating possible horizontal transfer of arsenic resistance genes in the soil bacterial community. Several isolates were able to reduce arsenate and to oxidise arsenite. In particular, Ancylobacter dichloromethanicum strain As3-1b possessed both characteristics, and arsenite oxidation occurred in the strain also under chemoautotrophic conditions. Some rhizobacteria produced siderophores, indole acetic acid and 1-amino-cyclopropane-1-carboxylic acid deaminase, thus possessing potential plant growth-promoting traits.

Arsenic-resistant bacteria isolated from agricultural soils of Bangladesh and characterization of arsenate-reducing strains

Aims:  To analyse the arsenic‐resistant bacterial communities of two agricultural soils of Bangladesh, to isolate arsenic‐resistant bacteria, to study their potential role in arsenic transformation and to investigate the genetic determinants for arsenic resistance among the isolates.

Microbial transformations of arsenic: perspectives for biological removal of arsenic from water

Arsenic is present in many environments and is released by various natural processes and anthropogenic actions. Although arsenic is recognized to cause a wide range of adverse health effects in humans, diverse bacteria can metabolize it by detoxification and energy conservation reactions. This review highlights the current understanding of the ecology, biochemistry and genomics of these bacteria, and their potential application in the treatment of arsenic-polluted water.

Distribution of catabolic pathways in some hydrocarbon-degrading bacteria from a subsurface polluted soil

Enrichment cultures on naphtha solvent were used to select aromatic hydrocarbon-degrading bacteria from a BTEX (benzene, toluene, ethylbenzene, xylene)-contaminated subsoil obtained from beneath a paint factory located in Milan, Italy. Fifteen isolated strains were studied for their different biodegradative capacities. Among these, 13 were able to grow on naphtha solvent. Ten were identified as Pseudomonas putida and three as Pseudomonas aureofaciens. Two other degraders were identified as Pseudomonas aeruginosa and Alcaligenes xylosoxidans subsp. denitrificans. Further molecular characterization of the isolates was carried out by randomly amplified polymorphic DNA analysis to ascertain that all the studied strains belonged to different haplotypes. The isolates were characterized for the presence of genes encoding for toluene dioxygenase, xylene monooxygenase and catechol 2,3-dioxygenase by polymerase chain reaction analysis and by Southern analysis. P. putida strain CM23, which showed homology with xylA,M, xylE and todC1C2BA genes, possessed multiple pathways which enabled the strain to grow on benzene, toluene and m-xylene.

Biotransformation of ferulic acid and related compounds by mutant strains of Pseudomonas fluorescens

Pseudomonas fluorescens strain FE2 isolated in the presence of ferulic acid was able to grow on hydroxylated and methoxylated compounds bearing the hydroxyl group in the para position. By ethylmethansulphonate (EMS) and transposon mutagenesis, mutants unable to utilize ferulic acid have been selected. The metabolic characterization of the wild-type strain and its mutants indicates that ferulic acid was degraded through the formation of vanillic acid. Mutant FE2B in co-oxidation experiments with glutamate, is able to transform ferulic and dihydroferulic acid into vanillic acid, 4-hydroxycinnamic acid and 3 (4-hydroxyphenyl)-propanoic acid into 4-hydroxybenzoic acid, and 3-hydroxycinnamic acid into 3-hydroxybenzoic acid. The bioconversion of hydroxylated aromatic substrates by the FE2B mutants suggests that the presence of a hydroxyl group on the aromatic ring is required for deacetylase activity.

Metabolism of ferulic acid by a facultatively anaerobic strain of Pseudomonas cepacia

Summary A facultatively anaerobic strain of Pseudomonas cepacia capable of utilizing aerobically ferulic acid as the only carbon and energy source was isolated. From the anaerobic Pseudomonas cultures supplied with ferulic acid and yeast extract, we isolated and identified 3-methoxy-4-hydroxystyrene and 3-methoxy-4-hydroxyphenylpropionic acid. These substances were not further metabolized. Aerobically growing cells completely metabolized ferulic acid through the intermediary formation of 3-methoxy-4-hydroxystyrene and 3-methoxy-4-hydroxyphenylpropionic acid.

Anaerobic digestion of winery wastewaters derived from different wine making processes

Abstract Anaerobic digestion of winery effluents derived from two different wine making processes was compared in a laboratory scale upflow filter. White winery effluents (WWE) were more easily degradable (average soluble COD removal=92%) than red winery effluents (RWE) (average soluble COD removal = 85%). Differences in the reactor microbial populations for the two effluents were also observed. Both aceticlastic and hydrogenotrophic methanogens in the circulating fractions were significantly higher during the treatment of WWE than during that of RWE. With both wastewaters, the reactor promptly reacted to volumetric organic load (Bv) stress (tripling from 4 to 12 g COD L‐1 d‐1). Following the Bv perturbation, COD removal decreased to 50%. However, after two weeks the COD removal values were as before perturbation. Volatile fatty acids, especially total VFAs and butyrate measured on the second day after the imposed Bv perturbation, were good indicators of process imbalance. Coenzyme F420 determination and ...

Arsenic transforming abilities of groundwater bacteria and the combined use of Aliihoeflea sp. strain 2WW and goethite in metalloid removal

Several technologies have been developed for lowering arsenic in drinking waters below the World Health Organization limit of 10 μg/L. When in the presence of the reduced form of inorganic arsenic, i.e. arsenite, one options is pre-oxidation of arsenite to arsenate and adsorption on iron-based materials. Microbial oxidation of arsenite is considered a sustainable alternative to the chemical oxidants. In this contest, the present study investigates arsenic redox transformation abilities of bacterial strains in reductive groundwater from Lombardia (Italy), where arsenite was the main arsenic species. Twenty isolates were able to reduce 75 mg/L arsenate to arsenite, and they were affiliated to the genera Pseudomonas, Achromobacter and Rhodococcus and genes of the ars operon were detected. Three arsenite oxidizing strains were isolated: they belonged to Rhodococcus sp., Achromobacter sp. and Aliihoeflea sp., and aioA genes for arsenite oxidase were detected in Aliihoeflea sp. strain 2WW and in Achromobacter sp. strain 1L. Uninduced resting cells of strain 2WW were used in combination with goethite for arsenic removal in a model system, in order to test the feasibility of an arsenic removal process. In the presence of 200 μg/L arsenite, the combined 2WW-goethite system removed 95% of arsenic, thus lowering it to 8 μg/L. These results indicate that arsenite oxidation by strain 2WW combined to goethite adsorption is a promising approach for arsenic removal from contaminated groundwater.