Papers by Cristina Becerra-Castro

The Science of the total environment, Jan 4, 2015
Some metals are nowadays considered environmental pollutants. Although some, like Cu and Zn, are ... more Some metals are nowadays considered environmental pollutants. Although some, like Cu and Zn, are essential for microorganisms, at high concentrations they can be toxic or exert selective pressures on bacteria. This study aimed to assess the potential of Cu or Zn as selectors of specific bacterial populations thriving in wastewater. Populations of Escherichia coli recovered on metal-free and metal-supplemented culture medium were compared based on antibiotic resistance phenotype and other traits. In addition, the bacterial groups enriched after successive transfers in metal-supplemented culture medium were identified. At a concentration of 1mM, Zn produced a stronger inhibitory effect than Cu on the culturability of Enterobacteriaceae. It was suggested that Zn selected populations with increased resistance prevalence to sulfamethoxazole or ciprofloxacin. In non-selective culture media, Zn or Cu selected for mono-species populations of ubiquitous Betaproteobacteria and Flavobacteriia,...

Environment International, 2015
The reuse of treated wastewater, in particular for irrigation, is an increasingly common practice... more The reuse of treated wastewater, in particular for irrigation, is an increasingly common practice, encouraged by governments and official entities worldwide. Irrigation with wastewater may have implications at two different levels: alter the physicochemical and microbiological properties of the soil and/or introduce and contribute to the accumulation of chemical and biological contaminants in soil. The first may affect soil productivity and fertility; the second may pose serious risks to the human and environmental health. The sustainable wastewater reuse in agriculture should prevent both types of effects, requiring a holistic and integrated risk assessment. In this article we critically review possible effects of irrigation with treated wastewater, with special emphasis on soil microbiota. The maintenance of a rich and diversified autochthonous soil microbiota and the use of treated wastewater with minimal levels of potential soil contaminants are proposed as sine qua non conditions to achieve a sustainable wastewater reuse for irrigation.
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ABSTRACT The aim of this study was to identify metal-tolerant plants with potential application i... more ABSTRACT The aim of this study was to identify metal-tolerant plants with potential application in phytoremediation strategies. For this we evaluated the distribution and chemical fractionation of heavy metals in soils and their accumulation or exclusion by native plant species growing in an abandoned Pb/Zn mine in NW Spain. Mine-soils (I = 0–20 cm, II = 20–40 cm) and shoots of the dominant plants were sampled at 39 sampling points in a (100 m × 100 m) regular grid at two sites (mine zone (MZ) and tailing ponds (TP)). Total metal contents in mine-soils presented a high heterogeneity and the principal contaminants were Cd, Hg, Pb and Zn (in agreement with the mineralogy of the site). Total concentrations varied from 2 to 95 mg Cd kg−1, 0.2 to 24.8 mg Hg kg−1, 46 to 6100 mg Pb kg−1, and 340 to 52,000 mg Zn kg−1. Soil metal distribution did not vary significantly with depth, but total and EDTA-extractable metal concentrations were higher in MZ than TP. On the other hand, the relative extractability of soil metals (EDTA-extractable concentration expressed as a % of total concentration) was higher in TP. The residual fraction was dominant for Co, Cr, Cu, and Ni, representing generally more than 50% of total concentration. Pb was preferentially associated with the oxidisable fraction, followed by the acid-extractable fraction. Both Cd and Zn were amply represented by the oxidisable, reducible and acid-extractable fractions. All the studied plant species were metal-tolerant (pseudometallophytes), although differences were found between species. Significant differences were found in plant accumulation of Cd and primarily, Zn. Cytisus scoparius and C. multiflorus were efficient metal-excluding species, and could be interesting candidates for phytostabilisation techniques and/or revegetation of heavily contaminated soils. Salix atrocinerea, presented significantly higher concentrations of Zn in its aerial biomass (543 ± 108 mg Zn kg−1) than the remaining species and bioconcentration factors (BCF) of up to 2.35 for Zn and 4.17 for Cd, respectively. This population could be interesting in the phytoextraction of low to moderately-contaminated soils.

Plant and Soil, 2013
ABSTRACT Background and aims Microbe-assisted phytoremediation is particularly effective for orga... more ABSTRACT Background and aims Microbe-assisted phytoremediation is particularly effective for organic pollutants. The leguminous shrub Cytisus striatus (Hill) Rothm. has been proposed as a candidate species for the rhizoremediation of hexachlorocyclohexane (HCH)-contaminated sites. The aim of this study was to improve the performance of this species using microbial inoculants. Methods C. striatus was grown in substrates contaminated with 0, 10 and 35 mg HCH kg−1 for 8 weeks. Plants were either not inoculated (NI), or inoculated with the endophyte Rhodococcus erythropolis ET54b and the HCH-degrader Sphingomonas sp. D4 (isolated from a HCH-contaminated soil) on their own or in combination (ET, D4 and ETD4). Results Inoculation with both bacterial strains (ETD4) resulted in decreased HCH phytotoxicity and improved plant growth. HCH-exposed plants inoculated with ETD4 presented a 120–160 % increase in root, and 140–160 % increase in shoot biomass, and led to a decrease in the activities of enzymes involved in anti-oxidative defence. APOD activity was reduced by up to 37 % in shoot tissues and 25 % in root tissues, and corresponding activities of SOD were reduced by up to 35 % and 30 %. HCH dissipation was enhanced in the presence of C. striatus but no significant effect of microbial inoculants was observed. Conclusions Inoculating C. striatus with this combination of bacterial strains is a promising approach for the remediation of HCH-contaminated sites.

Plant and Soil, 2014
ABSTRACT Aim Rhizobacteria can influence plant growth and metal accumulation. The aim of this stu... more ABSTRACT Aim Rhizobacteria can influence plant growth and metal accumulation. The aim of this study was to evaluate the effect of rhizobacterial inoculants on the Ni phytoextraction efficiency of the Ni-hyperaccumulator Alyssum pintodasilvae. Method In a preliminary screening 15 metal-tolerant bacterial strains were tested for their plant growth promoting (PGP) capacity or effect on Ni bioaccumulation. Strains were selected for their Ni tolerance, plant growth promoting traits and Ni solubilizing capacity. In a re-inoculation experiment five of the previously screened bacterial isolates were used to inoculate A. pintodasilvae in two contrasting Ni-rich soils (a serpentine (SP) soil and a sewage sludge-affected agricultural (LF) soil). Results Plant growth was greater in serpentine soil (where it grows naturally) than in the LF soil, probably due to Cd phytotoxicity. Rhizobacterial inoculants influenced plant growth and Ni uptake and accumulation, but the effect of the strains was dependent upon soil type. The increase in plant biomass and/or Ni accumulation significantly promoted shoot Ni removal. Conclusion One strain (Arthrobacter nicotinovorans SA40) was able to promote plant growth and phytoextraction of Ni in both soil types and could be a useful candidate for future field-based trials.

Environmental Pollution, 2013
The performance of Cytisus striatus in association with different microbial inoculant treatments ... more The performance of Cytisus striatus in association with different microbial inoculant treatments on the dissipation of the insecticide hexachlorocyclohexane (HCH) was studied. Two soils with different organic matter (A and B soil) content were spiked with 0 or 65 mg HCH kg(-1). Plants were either not inoculated (NI), or inoculated with the endophyte Rhodococcus erythropolis ET54b and the HCH-degrader Sphingomonas sp. D4 separately or in combination (ET, D4 and ETD4). Unplanted pots were also established. HCH phytotoxicity was more pronounced in the B soil. Soil HCH concentrations in unplanted pots were similar to initial concentrations, whereas concentrations were reduced after plant growth: by 20% and 8% in A and B soil, respectively. Microbial inoculants also modified HCH dissipation, although effects were soil-dependent. Inoculation with the combination of strains (ETD4) led to a significant enhancement in HCH dissipation: up to 53% in the A soil and 43% in the B soil.

International Journal of Phytoremediation
Bacterial strains were isolated from the rhizosphere of three populations of the Ni-hyperaccumula... more Bacterial strains were isolated from the rhizosphere of three populations of the Ni-hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum (A. pintodasilvae; M, S, and L), one population of Ni-hyperaccumulator A. serpyllifolium subsp. malacitanum (A. malacitanum; SB), and one population of the non-hyperaccumulator A. serpyllifolium subsp. serpyllifolium (A. serpyllifolium; SN). Isolates were characterized genotypically by BOX-PCR genomic DNA fingerprinting and comparative sequence analysis of partial 16S rRNA gene, and phenotypically by their Ni tolerance (0-10 mM), presence of plant growth promoting traits (indoleacetic acid (IAA)-, siderophore-, or organic acid-production, and phosphate solubilization) or capacity to produce biosurfactants. Among the collection of rhizobacteria, 84 strains were selected (according to their BOX-PCR profiles and phenotypic characteristics) to assess their ability to modify Ni extractability from Ni-rich (serpentine) soils. Metabolites produced b...

International journal of phytoremediation, Jan 1, 2011
Bacterial strains were isolated from the rhizosphere of three populations of the Ni-hyperaccumula... more Bacterial strains were isolated from the rhizosphere of three populations of the Ni-hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum (A. pintodasilvae; M, S, and L), one population of Ni-hyperaccumulator A. serpyllifolium subsp. malacitanum (A. malacitanum; SB), and one population of the non-hyperaccumulator A. serpyllifolium subsp. serpyllifolium (A. serpyllifolium; SN). Isolates were characterized genotypically by BOX-PCR genomic DNA fingerprinting and comparative sequence analysis of partial 16S rRNA gene, and phenotypically by their Ni tolerance (0-10 mM), presence of plant growth promoting traits (indoleacetic acid (IAA)-, siderophore-, or organic acid-production, and phosphate solubilization) or capacity to produce biosurfactants. Among the collection of rhizobacteria, 84 strains were selected (according to their BOX-PCR profiles and phenotypic characteristics) to assess their ability to modify Ni extractability from Ni-rich (serpentine) soils. Metabolites produced by 13 of the isolates mobilized soil Ni (originating from the rhizosphere of both Ni-hyperaccumulators and non-hyperaccumulator). In contrast, Ni extraction using culture medium filtrates which had supported the growth of 29 strains was significantly reduced. The remaining strains had no effect on Ni mobility. Bacterial induced Ni mobilization was not related to Ni resistance or the phenotypic traits tested. Isolates with potential use in phytoremediation techniques will be further studied in a plant-microorganism-soil system.

Journal of hazardous …, Jan 1, 2012
The plant-microorganism-soil system of three pseudometallophytes (Betula celtiberica, Cytisus sco... more The plant-microorganism-soil system of three pseudometallophytes (Betula celtiberica, Cytisus scoparius and Festuca rubra) growing in a Pb/Zn mine was characterised. Plant metal accumulation, soil metal fractions (rhizosphere and non-vegetated) and bacterial densities were determined. Total Cd, Pb and Zn in non-vegetated soils was up to 50, 3000 and 20,000 mg kg(-1) dry weight, respectively. The residual fraction dominated non-vegetated soils, whereas plant-available fractions became important in rhizosphere soils. All plant species effectively excluded metals from the shoot. F. rubra presented a shoot:root transport factor of ≤0.2 and this population could be useful in future phytostabilisation trials. Culturable bacterial densities and diversity were low (predominantly Actinobacteria). Rhizosphere soils hosted higher total and metal-tolerant bacterial densities. Seventy-four metal-tolerant rhizobacteria were isolated, and characterised genotypically (BOX-PCR, 16S rDNA) and phenotypically [Cd/Zn tolerance, biosurfactant production and plant growth promoting (PGP) traits]. Several isolates resisted high concentrations of Cd and Zn, and only a few presented PGP traits. Fourteen isolates were evaluated for promoting plant growth of two species (Salix viminalis and Festuca pratensis). Thirteen inoculants enhanced growth of F. pratensis, while only three enhanced growth of S. viminalis. Growth enhancement could not always be related to isolate PGP traits. In conclusion, some isolates show potential application in phytostabilisation or phytoextraction techniques.

International Journal of Phytoremediation, Jan 1, 2009
In this study we determine culturable microbial densities (total heterotrophs, ammonifiers, amylo... more In this study we determine culturable microbial densities (total heterotrophs, ammonifiers, amylolytics and cellulolytics) and bacterial resistance to Co, Cr, and Ni in bulk and rhizosphere soils of three populations of the Ni-hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum and the excluder Dactylis glomerata from ultramafic sites (two populations in Northeast (NE) Portugal (Samil (S), Morais (M)) and one population in Northwest (NW) Spain (Melide (L)). The relationship between bioavailable metal concentrations (H2O-soluble) and microbial densities were analysed. Significant differences in microbial densities and metal-resistance were observed between the two species and their three populations. The hyperaccumulator showed higher microbial densities (except cellulolytics) and a greater rhizosphere effect, but this was only observed in S and M populations. These populations of A. serpyllifolium also showed selective enrichment of Ni-tolerant bacteria at the rhizosphere where Ni solubility was enhanced (densities of Ni-resistant bacteria were positively correlated with H2O-soluble Ni). These rhizobacteria could solubilise Ni in the soil and potentially improve phytoextraction strategies.
Plant and soil, Jan 1, 2011

Ecosistemas, Jan 1, 2007
Aplicación de plantas hiperacumuladoras de níquel en la fitoextracción natural: el género Alyssum... more Aplicación de plantas hiperacumuladoras de níquel en la fitoextracción natural: el género Alyssum L. Las plantas metalofitas han desarrollado mecanismos biológicos que les permiten sobrevivir en suelos ricos en metales, tanto naturales (suelos serpentiníticos o ultramáficos) como antropogénicos. La mayoría consigue su tolerancia restringiendo fisiológicamente la entrada de metales a las raíces y/o el transporte hacia las hojas. Algunas especies, sin embargo, presentan mecanismos extremadamente especializados que les permiten acumular o "hiperacumular" metales (como Cd, Co, Ni y Zn) en sus hojas, hasta alcanzar concentraciones superiores al 2% de su materia seca: son las denominadas "plantas hiperacumuladoras". Las hiperacumuladoras de Ni son mucho más numerosas que las de otros metales. La mayoría de ellas pertenecen a la familia Brasicaceae, siendo Alyssum L. uno de los géneros más representados. La fitoextracción utiliza estas plantas para extraer los metales del suelo y acumularlos en la biomasa aérea. Tras su cosecha, los restos vegetales pueden ser reciclados o confinados de una forma poco costosa, lo que constituye una estrategia económica para la limpieza de suelos contaminados. Una importante limitación de la aplicación práctica de la mayoría de las especies hiperacumuladoras en la fitoextracción es su reducido tamaño y escasa biomasa. Optimizar las prácticas de manejo del suelo y la cosecha, con el objeto de incrementar su productividad y la concentración de metales en la biomasa, así como cultivar especies hiperacumuladoras mejoradas, podría ser, por tanto, una combinación clave en el desarrollo último de esta técnica.
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Papers by Cristina Becerra-Castro