Clementina Pozo
University of Granada, Microbiology, Faculty Member
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Research Interests: Biofilms, Biology, Filtration, Flavonoids, Medicine, and 15 moreEarth and Environmental Sciences, Biological Sciences, Anaerobic Digestion, Biofilm, Microbiology and Biotechnology, Bacteria, Degradation, Filter, Food Industry, Industrial Waste, Effluents, Chemical Oxygen Demand, Biofilter, Aeration, and Aromatic Compounds
Five strains of Rhizobium spp, one strain of Mesorhizobium loti and two strains of Sinorhizobium meliloti were tested for their ability to grow in chemically-defined medium lacking growth factor. Qualitative and quantitative production of... more
Five strains of Rhizobium spp, one strain of Mesorhizobium loti and two strains of Sinorhizobium meliloti were tested for their ability to grow in chemically-defined medium lacking growth factor. Qualitative and quantitative production of aspartic acid, serine, glutamic acid, glycine, histidine, threonine, arginine, alanine, proline, cysteine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine was determined by the use of mannitol as sole carbon source. Strains of Rhizobium spp. and Sinorhizobium sp. produced all the amino acids analysed with the exception of cysteine and high biological levels of serine, glycine and alanine were detected after 2 days of culture in mineral medium. Strain U226 of M. loti only produced small amounts of amino acids and glutamic acid, histidine, arginine, cysteine, methionine, lysine and phenylalanine was not liberated into the media.
Research Interests: Biochemistry, Carbon, Biology, Medicine, Rhizobium, and 15 moreTemperature, Amino Acids, Time Factors, Methionine, Amino Acid Profile, Sinorhizobium meliloti, Sinorhizobium, Glycine, Species Specificity, Growth Factor, Culture Media, Carbon Source, Glutamic Acid, Proteobacteria, and Medical biochemistry and metabolomics
Abstract The Aznalcollar accident, which occurred in 1998, spilled 36 × 105 m3 of pyritic sludge and 9 × 105 m3 of acidic water around an area of 43 km2 in the south of Spain. This spill is considered one of the most important... more
Abstract The Aznalcollar accident, which occurred in 1998, spilled 36 × 105 m3 of pyritic sludge and 9 × 105 m3 of acidic water around an area of 43 km2 in the south of Spain. This spill is considered one of the most important metal-mining associated accidents worldwide. In this study, two soil remediation techniques were evaluated: the addition of marble sludge (liming treatment, LS) and the mixing of recovered soils (RC) with contaminated soils (CT) (biopile treatment, BS). Both LS and BS significantly reduce the solubility of Cu, Zn, As, and Pb mainly due to the increase in pH and organic matter content, respectively. Soil basal respiration rate and the seed germination and root elongation bioassay with Lactuca sativa were used to evaluate the toxicity of the potential pollution in the sampled soils. Both bioassays showed that the CT soils exhibited the highest toxicity with a significant reduction in the toxicity of the amended soils (LS and BS). The abundance and structure of microbial communities in the soils were determined by qPCR and Illumina 16S rRNA sequencing, respectively. The absolute abundances of total bacterial and archaeal populations, ammonium oxidising bacteria, and denitrifiers in the CT soils were statistically lower than these found in the other three soils. Similarly, the structure of the bacterial community was highly different in the CT soils. Our results underline the persistence of the detrimental effect of pollutants in CT soils compared to the recuperated (RC) and amended soils (LS and BS). We also highlight the uses of liming or biopile as remediation techniques as satisfactory tools to reduce the impact of heavy metals in the contaminated Aznalcollar soils.
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Environmental Toxicology and Chemistry, Vol. 15, No. 7, pp. 11151118, 1996 ... 1996 SETAC Printed in the USA 0730-7268/96 $6.00 .00 ... STUDIES ON THE EFFECTS OF THE HERBICIDE SIMAZINE ON MICROFLORA OF FOUR AGRICULTURAL SOILS
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Abstract The goal of this research was to study the operation of an integrated fixed-film activated sludge system (IFAS), in anaerobic/anoxic/aerobic (A2O) configuration for the elimination of emerging pollutants, in particular of 27... more
Abstract The goal of this research was to study the operation of an integrated fixed-film activated sludge system (IFAS), in anaerobic/anoxic/aerobic (A2O) configuration for the elimination of emerging pollutants, in particular of 27 pharmaceutical active compounds (PhACs) (eight anti-inflammatory and/or analgesic pharmaceuticals (AIAPs), six antibiotics, four b-blockers, two antihypertensives/diuretics, three lipid regulators-bezafibrate and four psychiatric medications). Different operational conditions were analyzed in the biological reactor, controlling at all times both the amount of suspended and fixed biomass present. The A2O-IFAS system has obtained similar or higher removal efficiency (RE) in the elimination of organic matter, and especially of nutrients, nitrogen (N) and phosphorous (P), with respect to the conventional A2O system, working however with very low values of the sludge retention time (SRT) (4.0 days) and mix liquid suspended solid (MLSS) in the reactor (1822 mg L-1). In general, conventional activated sludge (CAS) and IFAS processes show the importance of operating at high SRT and MLSS concentration, that give low food/microorganisms (F/M) ratio, to increase the RE of several PhACs. However, the A2O-IFAS system, operating with low mixed liquor SRT and MLSS has achieved similar or better RE of PhACs, obtaining the highest average REs values (>80%) for fenofibrate, acetaminophen, ibuprofen, naproxen, clarithromycin and atenolol. According to the results obtained, due to the affordable cost and well-demonstrated performance, IFAS systems become one of the most promising technology for conventional wastewater treatment plants (WWTP) upgrading.
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Research Interests: Environmental Science, Microbial Ecology, Environmental Chemistry, Medicine, Chemical and Biological Engineering, and 15 moreBiological Sciences, Actinobacteria, Environmental Sciences, Cluster Analysis, Community Structure, CHEMICAL SCIENCES, Bacterial Diversity, Marine Sediment, Alphaproteobacteria, Anionic Surfactant, Agricultural Soil, Linear Alkylbenzene Sulfonate, Engineering System, DNA sequence, and Microcosm
Multi-contamination by organic pollutants and toxic metals is common in anthropogenic and industrial environments. In this study, the five fungal strains Chaetomium jodhpurense (MH667651.1), Chaetomium maderasense (MH665977.1),... more
Multi-contamination by organic pollutants and toxic metals is common in anthropogenic and industrial environments. In this study, the five fungal strains Chaetomium jodhpurense (MH667651.1), Chaetomium maderasense (MH665977.1), Paraconiothyrium variabile (MH667653.1), Emmia lacerata, and Phoma betae (MH667655.1), previously isolated in Tunisia, were investigated for the simultaneous removal and detoxification of phenanthrene (PHE) and benzo[a]anthracene (BAA), as well as heavy metals (HMs) (Cu, Zn, Pb and Ag) in Kirk’s media. The removal was analysed using HPLC, ultra-high performance liquid chromatography (UHPLC) coupled to a QToF mass spectrometer, transmission electron microscopy, and toxicology was assessed using phytotoxicity (Lepidium sativum seeds) and Microtox® (Allivibrio fisherii) assays. The PHE and BAA degradation rates, in free HMs cultures, reached 78.8% and 70.7%, respectively. However, the addition of HMs considerably affected the BAA degradation rate. The highest de...
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Abstract The Aznalcollar accident, which occurred in 1998, spilled 36 × 105 m3 of pyritic sludge and 9 × 105 m3 of acidic water around an area of 43 km2 in the south of Spain. This spill is considered one of the most important... more
Abstract The Aznalcollar accident, which occurred in 1998, spilled 36 × 105 m3 of pyritic sludge and 9 × 105 m3 of acidic water around an area of 43 km2 in the south of Spain. This spill is considered one of the most important metal-mining associated accidents worldwide. In this study, two soil remediation techniques were evaluated: the addition of marble sludge (liming treatment, LS) and the mixing of recovered soils (RC) with contaminated soils (CT) (biopile treatment, BS). Both LS and BS significantly reduce the solubility of Cu, Zn, As, and Pb mainly due to the increase in pH and organic matter content, respectively. Soil basal respiration rate and the seed germination and root elongation bioassay with Lactuca sativa were used to evaluate the toxicity of the potential pollution in the sampled soils. Both bioassays showed that the CT soils exhibited the highest toxicity with a significant reduction in the toxicity of the amended soils (LS and BS). The abundance and structure of microbial communities in the soils were determined by qPCR and Illumina 16S rRNA sequencing, respectively. The absolute abundances of total bacterial and archaeal populations, ammonium oxidising bacteria, and denitrifiers in the CT soils were statistically lower than these found in the other three soils. Similarly, the structure of the bacterial community was highly different in the CT soils. Our results underline the persistence of the detrimental effect of pollutants in CT soils compared to the recuperated (RC) and amended soils (LS and BS). We also highlight the uses of liming or biopile as remediation techniques as satisfactory tools to reduce the impact of heavy metals in the contaminated Aznalcollar soils.
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Four different laccase-producing strains were isolated from arid soils and used for bisphenol A (BPA) degradation. These strains were identified as Chaetomium strumarium G5I, Thielavia arenaria CH9, Thielavia arenaria HJ22 and Thielavia... more
Four different laccase-producing strains were isolated from arid soils and used for bisphenol A (BPA) degradation. These strains were identified as Chaetomium strumarium G5I, Thielavia arenaria CH9, Thielavia arenaria HJ22 and Thielavia arenaria SM1(III) by internal transcribed spacer 5.8 S rDNA analysis. Residual BPA was evaluated by HPLC analysis during 48 h of incubation. A complete removal of BPA was observed by the whole cell fungal cultures within different times, depending on each strain. C. strumarium G5I was the most efficient degrader, showing 100% of removal within 8 h of incubation. The degradation of BPA was accompanied by the production of laccase and dye decolorizing peroxidase (DyP) under degradation conditions. The presence of aminobenzotriazole (ABT) as an inhibitor of cytochrome P450s monooxygenases (CYP) demonstrated a slight decrease in BPA removal rate, suggesting the effective contribution of CYP in the conversion. The great involvement of laccase in BPA trans...
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A collection of twenty wild type strains of Rhizobium leguminosarum bv. viceae was obtained from five different agricultural locations in Granada (Spain) and tested with a commercial cultivar of faba bean (Vicia faba cv. Alborea) under... more
A collection of twenty wild type strains of Rhizobium leguminosarum bv. viceae was obtained from five different agricultural locations in Granada (Spain) and tested with a commercial cultivar of faba bean (Vicia faba cv. Alborea) under monoxenic conditions of culture in Leonard jars, in order to study the diversity of symbiotic efficiency among local isolates in terms of nodulation and
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Abstract Pharmaceutical active compounds (PhACs) are molecules used for the treatment of physiological disorders that can generate environmental imbalances when discharged into wastewater. In this work, the use of Penicillium oxalicum... more
Abstract Pharmaceutical active compounds (PhACs) are molecules used for the treatment of physiological disorders that can generate environmental imbalances when discharged into wastewater. In this work, the use of Penicillium oxalicum XD-3.1 as bioremediation tool for the elimination of PhACs in non-sterile real hospital wastewater in batch bench scale bioreactor and its impact on microbial community was evaluated. A quantitative analysis using UHPLC-QToF was performed under three different conditions to determine the elimination of the different (PhACs) by this ascomycetes fungus. On the other hand, the changes on microbial community over time were analysed using Illumina MiSeq sequencing platform and a phylogenetic microarray (PhyloChip). P. oxalicum XD-3.1 was able to reduce the concentration of the majority of the analysed PhACs in 24 h such as ketoprofen, naproxen and paracetamol. Moreover, P. oxalicum inhibited the native fungal populations present in the wastewater, including opportunistic human pathogens such as Mycosphaerella and Drechslera, as well as some bacterial human pathogens belonging to Clostridiaceae and Brucellaceae families. However, important degraders such as Pseudomonadaceae bacteria groups remained in the system, indicating a possible natural consortium formation between fungi and bacteria. These results indicate the possibility to use this system for the removal of PhACs in real applications.
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Emerging and unregulated contaminants end up in soils via stabilized/composted sewage sludges, paired with possible risks associated with the development of microbial resistance to antimicrobial agents or an imbalance in the microbial... more
Emerging and unregulated contaminants end up in soils via stabilized/composted sewage sludges, paired with possible risks associated with the development of microbial resistance to antimicrobial agents or an imbalance in the microbial communities. An enrichment experiment was performed, fortifying the sewage sludge with carbamazepine, ketoprofen and diclofenac as model compounds, with the aim to obtain strains with the capability to transform these pollutants. Culturable microorganisms were obtained at the end of the experiment. Among fungi, Cladosporium cladosporioides, Alternaria alternata and Penicillium raistrickii showed remarkable degradation rates. Population shifts in bacterial and fungal communities were also studied during the selective pressure using Illumina MiSeq. These analyses showed a predominance of Ascomycota (Dothideomycetes and Aspergillaceae) and Actinobacteria and Proteobacteria, suggesting the possibility of selecting native microorganisms to carry out bioreme...
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Non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics are two of the most employed drug groups around the world due to their use in the treatment of edema and pain. However, they also present an ecological challenge because they... more
Non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics are two of the most employed drug groups around the world due to their use in the treatment of edema and pain. However, they also present an ecological challenge because they are considered as potential water pollutants. In this work, the biodegradation of four NSAIDs (diclofenac, ibuprofen, naproxen and ketoprofen) and one analgesic (acetaminophen) at 50 µM (initial concentration) by Penicillium oxalicum, at both flask and bioreactor bench scales, was evaluated. An important co-metabolic mechanism as part of the global bioremediation process for the elimination of these drugs was observed, as in some cases it was necessary to supplement glucose to achieve a 100% removal rate: both individually and as a complex mixture. Identical behavior in the implementation of a fluidized bench-scale batch bioreactor, inoculated with pellets of this fungus and the complex mix of the drugs, was observed. The role of the cytochrome P450 ...
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Waste treatment and the simultaneous production of energy have gained great interest in the world. In the last decades, scientific efforts have focused largely on improving and developing sustainable bioprocess solutions for energy... more
Waste treatment and the simultaneous production of energy have gained great interest in the world. In the last decades, scientific efforts have focused largely on improving and developing sustainable bioprocess solutions for energy recovery from challenging waste. Anaerobic digestion (AD) has been developed as a low-cost organic waste treatment technology with a simple setup and relatively limited investment and operating costs. Different technologies such as one-stage and two-stage AD have been developed. The viability and performance of these technologies have been extensively reported, showing the supremacy of two-stage AD in terms of overall energy recovery from biomass under different substrates, temperatures, and pH conditions. However, a comprehensive review of the advantages and disadvantages of these technologies is still lacking. Since microbial ecology is critical to developing successful AD, many studies have shown the structure and dynamics of archaeal and bacterial com...
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Two microbial fuel cells were inoculated with activated sludge from Finland and operated under moderate (25 °C) and low (8 °C) temperatures. Operation under real urban wastewater showed similarities in chemical oxygen demand removal and... more
Two microbial fuel cells were inoculated with activated sludge from Finland and operated under moderate (25 °C) and low (8 °C) temperatures. Operation under real urban wastewater showed similarities in chemical oxygen demand removal and voltage generated, although moderate temperature supported higher ammonium oxidation. Fungi disappeared in the microbial fuel cell operated at temperature of 25 °C. Archaea domain was dominated by methanogenic archaea at both temperature scenarios. Important differences were observed in bacterial communities between both temperatures, however generating similar voltage. The results supported that the implementation of microbial fuel cells in Nordic countries operating under real conditions could be successful, as well as suggested the flexibility of cold-adapted inoculum for starting-up microbial fuel cells, regardless of the operating temperature of the system, obtaining higher COD removal and voltage generation performances at low temperature than ...
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Extractive membrane biofilm reactor (EMBFR) technology offers productive solutions for volatile and semi-volatile compound removal from water bodies. In this study, the bacterial strains Paenibacillus etheri SH7T (CECT 8558),... more
Extractive membrane biofilm reactor (EMBFR) technology offers productive solutions for volatile and semi-volatile compound removal from water bodies. In this study, the bacterial strains Paenibacillus etheri SH7T (CECT 8558), Agrobacterium sp. MS2 (CECT 8557) and Rhodococcus ruber strains A5 (CECT 8556), EE6 (CECT 8612) and EE1 (CECT 8555), previously isolated from fuel-contaminated sites, were tested for adherence on tubular semipermeable membranes in laboratory-scale systems designed for methyl tert-butyl ether (MTBE) bioremediation. Biofilm formation on the membrane surface was evaluated through observation by field-emission scanning electron microscope (FESEM) as well as the acute toxicity (as EC50) of the bacterial growth media. Moreover, extracellular polymeric substance (EPS) production for each strain under different MTBE concentrations was measured. Strains A5 and MS2 were biofilm producers and their adherence increased when the MTBE flowed through the inner tubular semiper...
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We report here the draft genome sequence ofPaenibacillus etherisp. nov. SH7T(= CECT 8558T= DSM 29760T), isolated from a hydrocarbon-contaminated soil pilot plant in Granada, Spain. The bacterium was isolated and sequenced due to its... more
We report here the draft genome sequence ofPaenibacillus etherisp. nov. SH7T(= CECT 8558T= DSM 29760T), isolated from a hydrocarbon-contaminated soil pilot plant in Granada, Spain. The bacterium was isolated and sequenced due to its methyl tert-butyl ether (MTBE)-degrading properties.
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Nine bacterial strains isolated from two hydrocarbon‐contaminated soils were selected because of their capacity for growth in culture media amended with 200 mg/L of one of the following gasoline oxygenates: Methyl‐tert‐butyl ether (MTBE),... more
Nine bacterial strains isolated from two hydrocarbon‐contaminated soils were selected because of their capacity for growth in culture media amended with 200 mg/L of one of the following gasoline oxygenates: Methyl‐tert‐butyl ether (MTBE), ethyl‐tert‐butyl ether (ETBE), and tert‐amyl methyl ether (TAME). These strains were identified by amplification of their 16S rRNA gene, using fD1 and rD1 primers, and were tested for their capacity to grow and biotransform these oxygenates in both mineral and cometabolic media. The isolates were classified as Bacillus simplex, Bacillus drentensis, Arthrobacter sp., Acinetobacter calcoaceticus, Acinetobacter sp., Gordonia amicalis (two strains), Nocardioides sp., and Rhodococcus ruber. Arthrobacter sp. (strain MG) and A. calcoaceticus (strain M10) consumed 100 (cometabolic medium) and 82 mg/L (mineral medium) of oxygenate TAME in 21 d, respectively, under aerobic conditions. Rhodococcus ruber (strain E10) was observed to use MTBE and ETBE as the so...
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Emerging water contaminants derived from unleaded gasoline such as methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE) and tert-amyl methyl ether (TAME), are in need of effective bioremediation technologies for restoring water... more
Emerging water contaminants derived from unleaded gasoline such as methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE) and tert-amyl methyl ether (TAME), are in need of effective bioremediation technologies for restoring water resources. In order to design the conditions of a future groundwater bioremediating biofilter, this work assesses the potential use of Acinetobacter calcoaceticus M10, Rhodococcus ruber E10 and Gordonia amicalis T3 for the removal of MTBE, ETBE and TAME in consortia or as individual strains. Biofilm formation on an inert polyethylene support material was assessed with scanning electron microscopy, and consortia were also analysed with fluorescent in situ hybridisation to examine the relation between the strains. A. calcoaceticus M10 was the best coloniser, followed by G. amicalis T3, however, biofilm formation of pair consortia favoured consortium M10-E10 both in formation and activity. However, degradation batch studies determined that neither consortium exhibited higher degradation than individual strain degradation. The physiological state of the three strains was also determined through flow cytometry using propidium iodide and 3'-dihexylocarbocyanine iodide thus gathering information on their viability and activity with the three oxygenates since previous microbial counts revealed slow growth. Strain E10 was observed to have the highest physiological activity in the presence of MTBE, and strain M10 activity with TAME was only maintained for 24 h, thus we believe that biotransformation of MTBE occurs within the active periods established by the cytometry analyses. Viable cell counts and oxygenate removal were determined in the presence of the metabolites tert-butyl alcohol (TBA) and tert-amyl alcohol (TAA), resulting in TBA biotransformation by M10 and E10, and TAA by M10. Our results show that A. calcoaceticus M10 and the consortium M10-E10 could be adequate inocula in MTBE and TAME bioremediating technologies.