Magdalena Grifoll
Universitat de Barcelona, Microbiology, Faculty Member
Poster presentado en el International Meeting on New Strategies in Bioremediation Processes BioRemid-2017, March 9th-10th, (2017)
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Aerobic degradation in the laboratory of C 11 C 14 linear alkylbenzenes by pure cultures of bacterial strains ( ) revealed that biodegradation of individual isomers increases when the phenyl group is closer to the end of the alkyl chain.... more
Aerobic degradation in the laboratory of C 11 C 14 linear alkylbenzenes by pure cultures of bacterial strains ( ) revealed that biodegradation of individual isomers increases when the phenyl group is closer to the end of the alkyl chain. This result contributes to the understanding of ...
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The application of Salmonella/microsomal mammalian tests to column chromatography fractions isolated from river and marine sediments collected in the vicinity of Barcelona city, Spain, demonstrated a positive response (TA98 + S9 mix)... more
The application of Salmonella/microsomal mammalian tests to column chromatography fractions isolated from river and marine sediments collected in the vicinity of Barcelona city, Spain, demonstrated a positive response (TA98 + S9 mix) among the polar fractions. Chemical analysis by high resolution gas chromatography coupled to negative ion chemical ionization mass spectrometry (HRGC-NICI MS) provided sensitivity and selectivity to detect several mutagenic chemical classes. Among them, nitrated PAHs, azaarenes, aromatic amines, anhydrides, and ketones were identified. A total of 116 compounds were tentatively identified, 22 for the first time, of which 16 possessed mutagenic activity. However, a lack of correlation between chemical composition and fraction mutagenicity in the medium polarity fractions, especially in the river sediment, was evidenced. The occurrence of multiple interactions between components in spiked organic extracts is demonstrated.
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Research Interests: Chemistry, Environmental Pollution, Multidisciplinary, Applied Environmental Microbiology, Pseudomonas, and 12 moreDibenzothiophene, Gas Chromatography, Polycyclic Aromatic Hydrocarbon, Polycyclic aromatic hydrocarbons (PAHs), Gas Chromatography/mass Spectrometry, Environmental Pollutants, Phenanthrene, Acenaphthene, Naphthalene, Fluorene, Oxidation-Reduction, and Molecular Structure
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Particulate (greater than 0.22 microns) and dissolved phases of water concentrates (600 mL) of Llobregat and Besos Rivers (Barcelona, Spain), were tested in the Salmonella/microsome assay, tester strains TA98 and TA100. Most of them... more
Particulate (greater than 0.22 microns) and dissolved phases of water concentrates (600 mL) of Llobregat and Besos Rivers (Barcelona, Spain), were tested in the Salmonella/microsome assay, tester strains TA98 and TA100. Most of them showed significant mutagenic activity. However, independently of the application of exogenous metabolic activation, the dimethylsulfoxide extracts of the particulate matter exhibited a stronger mutagenic activity than the dissolved phase. This indicated that both rivers are chronically polluted by frameshift and base-pair substitution mutagens and promutagens. In order to investigate their identity, a bioassay-directed column chromatography fractionation of the base-neutrals isolated from the dissolved and particulate phases of Besos river water (7 L) was carried out. The mutagenic activity (TA98) was higher in presence of S9 and was recovered in the more polar fractions, where several mutagenic agents were identified by capillary GC-MS in the negative ion chemical ionization mode (NICI). Among them, o-tolidine, nitroquinoline, nitroaniline, dichlorobenzidine and several aromatic quinones were candidates for fraction mutagenicity.
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Research Interests: Evolutionary Biology, Microbiology, Environmental microbiology, Carbon, Biology, and 15 moreMedicine, Fatty acids, Intact polar lipids, Cluster Analysis, Catalase, Genus, Alphaproteobacteria, Fatty Acid, Environmental Pollutants, New Genus, Culture Media, Biological Macromolecules, Carbon Source, Fuel Oils, and Molecular Sequence Data
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Research Interests: Chemistry, Kinetics, Applied microbiology, Multidisciplinary, Biodegradation, and 10 moreMycobacterium, Polycyclic Aromatic Hydrocarbon, Applied Microbiology and Biotechnology, Polycyclic aromatic hydrocarbons (PAHs), Analytical Method, Time Factors, Gas Chromatography/mass Spectrometry, Culture Media, Pyrene, and Oxidation-Reduction
Polar biotransformation products have been identified as causative agents for the eventual increase in genotoxicity observed after the bioremediation of PAH-contaminated soils. Their further biodegradation has been described under certain... more
Polar biotransformation products have been identified as causative agents for the eventual increase in genotoxicity observed after the bioremediation of PAH-contaminated soils. Their further biodegradation has been described under certain biostimulation conditions; however, the underlying microorganisms and mechanisms remain to be elucidated. 9,10-Anthraquinone (ANTQ), a transformation product from anthracene (ANT), is the most commonly detected oxygenated PAH (oxy-PAH) in contaminated soils. Sand-in-liquid microcosms inoculated with creosote-contaminated soil revealed the existence of a specialized ANTQ degrading community, and Sphingobium sp. AntQ-1 was isolated for its ability to grow on this oxy-PAH. Combining the metabolomic, genomic, and transcriptomic analyses of strain AntQ-1, we comprehensively reconstructed the ANTQ biodegradation pathway. Novel mechanisms for polyaromatic compound degradation were revealed, involving the cleavage of the central ring catalyzed by Baeyer–Villiger monooxygenases (BVMO). Abundance of strain AntQ-1 16S rRNA and its BVMO genes in the sand-in-liquid microcosms correlated with maximum ANTQ biodegradation rates, supporting the environmental relevance of this mechanism. Our results demonstrate the existence of highly specialized microbial communities in contaminated soils responsible for processing oxy-PAHs accumulated by primary degraders. Also, they underscore the key role that BVMO may play as a detoxification mechanism to mitigate the risk posed by oxy-PAH formation during bioremediation of PAH-contaminated soils.
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An Arthrobacter sp. strain, F101, able to use fluorene as the sole source of carbon and energy, was isolated from sludge from an oil refinery wastewater treatment plant. During growth in the presence of fluorene, four major metabolites... more
An Arthrobacter sp. strain, F101, able to use fluorene as the sole source of carbon and energy, was isolated from sludge from an oil refinery wastewater treatment plant. During growth in the presence of fluorene, four major metabolites were detected and isolated by thin-layer chromatography and high-performance liquid chromatography. 9-Fluorenol, 9H-fluoren-9-one, and 3,4-dihydrocoumarin were identified by UV spectra, mass spectrometry, and 300-MHz proton nuclear magnetic resonance. The fourth metabolite has been characterized, but precise identification was not possible. Since strain F101 is not able to grow with fluorenone, two different pathways of fluorene biodegradation are suggested: one supports cell growth and produces 3,4-dihydrocoumarin as an intermediate and probably the unidentified metabolite, and the other produces 9-fluorenol and 9H-fluoren-9-one and appears to be a dead-end route.
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A two-stage Partial Nitritation (PN)/Anammox process was carried out at lab-scale conditions to treat reject water from a municipal WWTP. PN was achieved in a granular SBR obtaining an effluent with a NH4(+)-N/NO2(-)-N molar ratio around... more
A two-stage Partial Nitritation (PN)/Anammox process was carried out at lab-scale conditions to treat reject water from a municipal WWTP. PN was achieved in a granular SBR obtaining an effluent with a NH4(+)-N/NO2(-)-N molar ratio around 1.0. The microbial characterization of this reactor revealed a predominance of Betaproteobacteria, with a member of Nitrosomonas as the main autotrophic ammonium oxidizing bacterium (AOB). Nitrite oxidizing bacteria (NOB) were under the detection limit of 16S rRNA gene pyrosequencing, indicating their effective inhibition. The effluent of the PN reactor was fed to an Anammox SBR where stable operation was achieved with a NH4(+)-N:NO2(-)-N:NO3(-)-N stoichiometry of 1:1.25:0.14. The deviation to the theoretical stoichiometry could be attributed to the presence of heterotrophic biomass in the Anammox reactor (mainly members of Chlorobi and Chloroflexi). Planctomycetes accounted for 7% of the global community, being members of Brocadia (1.4% of the total abundance) the main anaerobic ammonium oxidizer detected.
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Research Interests: Evolutionary Biology, Microbiology, Environmental microbiology, Carbon, Biology, and 15 moreMedicine, Fatty acids, Intact polar lipids, Cluster Analysis, Catalase, Genus, Alphaproteobacteria, Fatty Acid, Environmental Pollutants, New Genus, Culture Media, Biological Macromolecules, Carbon Source, Fuel Oils, and Molecular Sequence Data
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Research Interests: Biology, Environmental Chemistry, Environmental Pollution, Medicine, Biological Sciences, and 15 morePhylogeny, Environmental Sciences, Hydrocarbon, Bacteria, Community Structure, Anthracene, Polycyclic Aromatic Hydrocarbon, Seawater, Water Microbiology, Phenanthrene, Pyrene, Fluoranthene, Microbial Community Structure, Fuel Oils, and Medical and Health Sciences
Research Interests: Microbiology, Chemistry, Soil, Kinetics, Bioremediation, and 15 moreEnvironmental Chemistry, Medicine, Biodegradation, Soil contamination, Mycobacterium, Polycyclic Aromatic Hydrocarbon, Polycyclic aromatic hydrocarbons (PAHs), Phenanthrene, Biological activity, Soil Water, Metabolite, Agricultural Soil, Phthalic acid, High Molecular Weight, and Microcosm
Marine microbial consortium UBF, enriched from a beach polluted by the Prestige oil spill and highly efficient in degrading this heavy fuel, was subcultured in pyrene minimal medium. The pyrene-degrading subpopulation (UBF-Py) mineralized... more
Marine microbial consortium UBF, enriched from a beach polluted by the Prestige oil spill and highly efficient in degrading this heavy fuel, was subcultured in pyrene minimal medium. The pyrene-degrading subpopulation (UBF-Py) mineralized 31 % of pyrene without accumulation of partially oxidized intermediates indicating the cooperation of different microbial components in substrate mineralization. The microbial community composition was characterized by culture dependent and PCR based methods (PCR-DGGE and clone libraries). Molecular analyses showed a highly stable community composed by Alphaproteobacteria (84 %, Breoghania, Thalassospira, Paracoccus, and Martelella) and Actinobacteria (16 %, Gordonia). The members of Thalasosspira and Gordonia were not recovered as pure cultures, but five additional strains, not detected in the molecular analysis, that classified within the genera Novosphingobium, Sphingopyxis, Aurantimonas (Alphaproteobacteria), Alcanivorax (Gammaproteobacteria) and Micrococcus (Actinobacteria), were isolated. None of the isolates degraded pyrene or other PAHs in pure culture. PCR amplification of Gram-positive and Gram-negative dioxygenase genes did not produce results with any of the cultured strains. However, sequences related to the NidA3 pyrene dioxygenase present in mycobacterial strains were detected in UBF-Py consortium, suggesting the representative of Gordonia as the key pyrene degrader, which is consistent with a preeminent role of actinobacteria in pyrene removal in coastal environments affected by marine oil spills.
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The pyrene-degrading Mycobacterium sp. strain AP1 grew in nutrient-supplemented artificial seawater with a heavy fuel oil as the sole carbon source, causing the complete removal of all linear (C 12 to C 40 ) and branched alkanes from the... more
The pyrene-degrading Mycobacterium sp. strain AP1 grew in nutrient-supplemented artificial seawater with a heavy fuel oil as the sole carbon source, causing the complete removal of all linear (C 12 to C 40 ) and branched alkanes from the aliphatic fraction, as well as an extensive degradation of the three- and four-ring polycyclic aromatic hydrocarbons (PAHs) phenanthrene (95%), anthracene (80%), fluoranthene (80%), pyrene (75%), and benzo( a )anthracene (30%). Alkylated PAHs, which are more abundant in crude oils than the nonsubstituted compounds, were selectively attacked at extents that varied from more than 90% for dimethylnaphthalenes, methylphenanthrenes, methylfluorenes, and methyldibenzothiophenes to about 30% for monomethylated fluoranthenes/pyrenes and trimethylated phenanthrenes and dibenzothiophenes. Identification of key metabolites indicated the utilization of phenanthrene, pyrene, and fluoranthene by known assimilatory metabolic routes, while other components were coo...
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A novel biphasic system containing mineral medium and sand coated with a biologically weathered creosote-PAH mixture was developed to specifically enrich the high molecular weight polycyclic aromatic hydrocarbon (HMW PAH)-degrading... more
A novel biphasic system containing mineral medium and sand coated with a biologically weathered creosote-PAH mixture was developed to specifically enrich the high molecular weight polycyclic aromatic hydrocarbon (HMW PAH)-degrading community from a creosote-polluted soil. This consortium (UBHP) removed 70 % of the total HMW PAHs and their alkyl-derivatives in 12 weeks. Based on a combined culture-dependent/independent approach, including clone library analysis, detection of catabolic genes, metabolomic profiles, and characterization of bacterial isolates, 10 phylotypes corresponding to five major genera (Sphingobium, Sphingomonas, Achromobacter, Pseudomonas, and Mycobacterium) were pointed out as key players within the community. In response to exposure to different single PAHs, members of sphingomonads were associated to the utilization of phenanthrene, fluoranthene, benzo[a]anthracene, and chrysene, while the degradation of pyrene was mainly associated to low-abundance mycobacteri...
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Phytoremediation of organic contaminants mainly depends on plant-microbe interactions in the rhizosphere, where, plant roots stimulate microbial populations for enhanced the biodegradation in polluted soil. We conducted a green-house... more
Phytoremediation of organic contaminants mainly depends on plant-microbe interactions in the rhizosphere, where, plant roots stimulate microbial populations for enhanced the biodegradation in polluted soil. We conducted a green-house experiment to measure dissipation of polycyclic aromatic hydrocarbons (PAHs) in the rhizosphere of sunflower roots on a creosote-polluted soil (containing 21.75 mg·Kg-1 of 6 PAHs), and an in vitro production of root exudates with 50 seeds of sunflower cv. in sterile conditions in an inorganic salts solution used in mineralization experiments (MM; pH=5,7) to study its effect on the biodegradation of phenanthrene and pyrene (present in creosote). In this way, we may separate the chemical impact of the root exudates from any root surface phenomena indicating that sunflower root exudates have the potential to increase the degradation of xenobiotics by the growth of soil microorganisms. In addition, we performed a characterization of these exudates of sunflo...
A fluorene-utilizing microorganism, identified as a species of Pseudomonas, was isolated from soil severely contaminated from creosote use and was shown to accumulate six major metabolites from fluorene in washed-cell incubations. Five of... more
A fluorene-utilizing microorganism, identified as a species of Pseudomonas, was isolated from soil severely contaminated from creosote use and was shown to accumulate six major metabolites from fluorene in washed-cell incubations. Five of these products were identified as 9-fluorenol, 9-fluorenone, (+)-1,1a-dihydroxy-1-hydro-9-fluorenone, 8-hydroxy-3,4-benzocoumarin, and phthalic acid. This last compound was also identified in growing cultures supported by fluorene. Fluorene assimilation into cell biomass was estimated to be approximately 50%. The structures of accumulated products indicate that a previously undescribed pathway of fluorene catabolism is employed by Pseudomonas sp. strain F274. This pathway involves oxygenation of fluorene at C-9 to give 9-fluorenol, which is then dehydrogenated to the corresponding ketone, 9-fluorenone. Dioxygenase attack on 9-fluorenone adjacent to the carbonyl group gives an angular diol, 1,1a-dihydroxy-1-hydro-9-fluorenone. Identification of 8-hy...
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Cycling of pollutants is essential to preserve functional marine and terrestrial ecosystems. Progress in optimizing these natural biological processes relies on the identification of the underlying microbial actors and deciphering their... more
Cycling of pollutants is essential to preserve functional marine and terrestrial ecosystems. Progress in optimizing these natural biological processes relies on the identification of the underlying microbial actors and deciphering their interactions at molecular, cellular, community, and ecosystem level. Novel advances on PAH biodegradation are built on a progressive approach that span from pure cultures to environmental communities, illustrating the complex metabolic networks within a single cell, and their further implications in higher complexity systems. Recent analytical chemistry and molecular tools allow a deeper insight into the active microbial processes actually occurring in situ, identifying active functions, metabolic pathways and key players. Understanding these processes will provide new tools to assess biodegradation occurrence and, as a final outcome, predict the success of bioremediation thus reducing its uncertainties, the main drawback of this environmental biotechnology.
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ABSTRACT
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ABSTRACT Oxygenated polyaromatic hydrocarbons (oxy-PAHs) and nitrogen heterocyclic polycyclic aromatic compounds (N-PACs) are toxic, highly leachable and often abundant at sites that are also contaminated with a PAH. However, due to lack... more
ABSTRACT Oxygenated polyaromatic hydrocarbons (oxy-PAHs) and nitrogen heterocyclic polycyclic aromatic compounds (N-PACs) are toxic, highly leachable and often abundant at sites that are also contaminated with a PAH. However, due to lack of regulations and standardized methods for their analysis, they are seldom included in monitoring and risk-assessment programs. This intercomparison study constitutes an important step in harmonizing the analytical methods currently used, and may also be considered a first step towards certifying reference materials for these compounds. The results showed that the participants were able to determine oxy-PAHs with accuracy similar to PAHs, with average determined mass fractions agreeing well with the known levels in a spiked soil and acceptable inter- and intra- laboratory precisions for all soils analyzed. For the N-PACs, the results were less satisfactory, and have to be improved by using analytical methods optimized for these compounds in a more tailored way.