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Research Project

RESTORATION OF ABANDONED MINING SITES

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Heavy metal accumulation in plant species indigenous to a contaminated portuguese site: prospects for phytoremediation
Publication . Moreira, Helena; Marques, Ana P. G. C.; Rangel, António O. S. S.; Castro, Paula M. L.
Phytoremediation is a promising alternative to conventional soil clean-up methods; however, up to date, there is still not enough information on plant species suitable for application in this field of science. Therefore, plant screening on contaminated sites can lead to the identification of further species of interest. In the present study, pedological and botanical characteristics of an industrialised area known for its metal contamination, in special with Zn—Esteiro de Estarreja, in Portugal—were examined in a 1-year screening. Twenty-seven species were found, with a higher occurrence and variability in the summer/spring season. Zinc levels in the tissues of the collected plant samples ranged from 34 mg kg−1 in shoots to 2,440 mg kg−1 in roots of different species. Species as Verbascum virgatum, Hypochoeris radicata, Phalaris arundinacea, Conyza bilbaoana, Paspalum urvillei and Aster squamatus have shown high Zn shoot accumulation and bioconcentration factors (BCFshoots > 1) and high metal translocation factors (TF > 1). Others, namely Spergularia capillacea, excluded Zn from the shoot tissues and stored the metal at the root zone (BCFroots > 1), behaving as tolerant plants. Plants were also screened for arbuscular mycorrhizal fungi colonisation, and only few species showed mycorrhizal presence, namely C. bilbaoana, Hirschfeldia incana, Epilobium tetragonum, Conyza sumatrensis, Pteridium aquilinum, P. urvillei and A. squamatus. The present work showed important indigenous species that can cope with installed harsh conditions and with potential for utilisation in phytoremediation strategies, either through metal removal to aerial parts or through its immobilisation in the root zone.
Assessment of the plant growth promotion abilities of six bacterial isolates using Zea mays as indicator plant
Publication . Marques, Ana P. G. C.; Pires, Carlos; Moreira, Helena; Rangel, António O. S. S.; Castro, Paula M. L.
Zea mays, one of the most important cereals worldwide, is a plant not only with food and energy value, but also with phytoremediation potential. The use of plant growth promoting (PGP) rhizobacteria may constitute a biological alternative to increase crop yield and plant resistance to degraded environments. In search for PGP rhizobacteria strains, 6 bacterial isolates were isolated from a metal contaminated site, screened in vitro for their PGP characteristics and their effects on the growth of Z. mays were assessed. Isolates were identified as 3A10T, ECP37T, corresponding to Chryseobacterium palustre and Chryseobacterium humi, and 1ZP4, EC15, EC30 and 1C2, corresponding to strains within the genera Sphingobacterium, Bacillus, Achromobacter, and Ralstonia, respectively. All the bacterial isolates were shown to produce indole acetic acid, hydrogen cyanide and ammonia when tested in vitro for their plant growth promoting abilities, but only isolates 1C2, 1ZP4 and ECP37T have shown siderophore production. Their further application in a greenhouse experiment using Z. mays indicated that plant traits such as root and shoot elongation and biomass production, and nutrient status, namely N and P levels, were influenced by the inoculation, with plants inoculated with 1C2 generally outperforming the other treatments. Two other bacterial isolates, 1ZP4 and ECP37T also led to increased plant growth in the greenhouse. These 3 species, corresponding to strains within the genera Ralstonia (1C2), Sphingobacterium (1ZP4), and to a strain identified as C. humi (ECP37T) can thus be potential agents to increase crop yield in maize plants.
Bacteria immobilisation on hydroxyapatite surface for heavy metals removal
Publication . Piccirillo, C.; Pereira, S. I. A.; Marques, A. P. G. C.; Pullar, R. C.; Tobaldi, D. M.; Pintado, M. E.; Castro, P. M. L.
Selected bacterial strains were immobilised on the surface of hydroxyapatite (Ca-10(PO4)(6)(OH)(2) - HAp) of natural origin (fish bones). The capacity of the material, alone and in combination with the bacterial strains to act as heavy metal removers from aqueous streams was assessed. Pseudomonas fluorescens (S3X), Micro bacterium oxydans (EC29) and Cupriavidus sp. (1C2) were chosen based on their resistance to heavy metals and capacity of adsorbing the metals. These systems were tested using solutions of Zn(II), Cd(II) and in solutions containing both metals. A synergistic effect between the strains and HAp, which is effective in removing the target heavy metals on its own, was observed, as the combination of HAp with the bacterial strains led to higher adsorption capacity for both elements. For the solutions containing only one metal the synergistic effect was greater for higher metal concentrations; 1C2 and EC29 were the most effective strains for Zn(II) and Cd(II) respectively, while S3X was less effective. Overall, an almost four-fold increase was observed for the maximum adsorption capacity for Zn(II) when 10 was employed - 0.433 mmol/g in comparison of 0.121 mmol/g for the unmodified HAP. For Cd(II), on the other hand, an almost three-fold increase was registered with EC29 bacterial strain - 0.090 vs 0.036 mmol/g for the unmodified HAp. When the solutions containing both metals were tested, the effect was more marked for lower concentrations.
The effect of ectomycorrhizal fungi forming symbiosis with Pinus pinaster seedlings exposed to cadmium
Publication . Sousa, Nadine R.; Ramos, Miguel A.; Marques, Ana P. G. C.; Castro, Paula M. L.
Cadmium is one of the most toxic heavy metals and its accumulation in the upper layers of forest soils affects plants, microorganisms and their interactions. Adequate strategies for the reforestation of metal contaminated sites are of vital importance. The aim of this work was to evaluate the response of Pinus pinaster seedlings to Cd exposure and to assess the effect of inoculation with two selected ectomycorrhizal fungi, Suillus bovinus and Rhizopogon roseolus on that response. Seedlings were exposed to soil contaminated at 15 and 30 mg Cd kg−1. Shoot biomass of P. pinaster decreased ca. 36% when exposed to 15 mg Cd kg−1. Overall, colonization by S. bovinus significantly enhanced shoot development up to 30% in contaminated soil while colonization by R. roseolus produced no significant effect at both Cd concentrations tested and significantly increased the level of Cd in the shoots at both Cd concentrations. Metal accumulation in the shoots and roots of non-inoculated and S. bovinus-inoculated seedlings increased at the higher Cd levels whereas R. roseolus-inoculated seedlings were not sensitive to Cd variation in the soil. The results from our research show that inoculation with ECM fungi has a significant impact on metal uptake and development of P. pinaster seedlings; the differential response induced by the two tested species highlights the importance of selecting the appropriate strains for nursery inoculation, and, as such, this biological tool ought to be considered in reforestation processes of heavy metal contaminated areas by woody species.
Performance of an aerobic granular sequencing batch reactor fed with wastewaters contaminated with Zn2+
Publication . Marques, Ana P. G. C.; Duque, Anouk F.; Bessa, Vânia S.; Mesquita, Raquel B. R.; Rangel, António O. S. S.; Castro, Paula M. L.
The main aim of this study was to investigate the performance of an aerobic granular sludge sequencing batch reactor (AGS-SBR) receiving water streams supplied with different loads of Zn2+ (50 and 100 mg L-1) during an operation of 866 cycles (ca. 109 days). When the metal was not fed, chemical oxygen demand (COD), PO43- and NH4+ were efficiently removed, with efficiencies of 56, 23 and 72% respectively. DGGE profiles showed that Zn2+ supply negatively affected the bacterial diversity and community structure of the granules. Consequently, the shock loadings with Zn2+, particularly at the higher levels (100 mg L-1), affected the nutrient removal in the AGS-SBR, although the reactor still generally complied with admissible legal values concerning organic matter, nitrogen and Zn. Simultaneous removal of PO43- and TSS in such conditions needs further refining but the application of aerobic granular SBR in the treatment of Zn2+ contaminated wastewaters seems viable.

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Funding agency

Fundação para a Ciência e a Tecnologia

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Funding Award Number

SFRH/BPD/34585/2007

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