Ana P. Mucha

Macrobenthic community in the Douro estuary: relations with trace metals and natural sediment characteristics.

The relationship between macrobenthic community structure and natural characteristics of sediment and trace metal contamination were studied in the lower Douro estuary (Portugal, NW, Iberian Peninsula), using an innovative threefold approach (SQG, Sediment Quality Guidelines), metal normalization to Fe, and macrobenthic community structure. This study allowed detection of a clear signature of anthropogenic contamination, in terms of Zn, Cu, Pb, and Cr in the north bank of the estuary, which experiences high urban pressure. Using the SQG approach, metal concentrations above ERM (effects range-median) were observed only at one sampling station, but several stations had levels above ERL (effects range-low). The macrobenthic community had a low diversity, with only 19 species found in the entire estuarine area, dominated by opportunistic species. The granulometric distribution of the sediments (estimated from the combination of organic matter, Fe and Al) seemed to be the major structuring factor for the communities, establishing the natural macrobenthic distribution pattern. The metals (Zn, Cu, Pb, and Cr) seemed to act as a disturbing factor over the natural distribution, with deleterious consequences for the macrobenthic communities.

Can PAHs influence Cu accumulation by salt marsh plants

The presence of polycyclic aromatic hydrocarbons (PAHs) may change the mechanisms of metal uptake, thus influencing kinetics and extent of metal phytoextraction. Studies on the subject are scarce, particularly for salt marsh plants. The aim of this work was to investigate the effect of PAHs on the uptake of Cu by Halimione portulacoides, a plant commonly found in salt marshes. Experiments were carried out in the laboratory, either in hydroponics (sediment elutriate) or in sediment soaked in elutriate, which were prepared with sediment and water from a salt marsh of the Cavado river estuary (NW Portugal). Groups of H. portulacoides (grown in a greenhouse) were exposed to those media during six days. Cu2+ (as Cu(NO3)2), 10(2) and 10(4) microg l(-1), was added to the media as well as 1.6 microg l(-1) of the sixteen EPA priority PAHs (0.1 microg l(-1) of each PAHs). Cu was assayed in solutions, sediments and different plant tissues before and after experiments. After exposure, photosynthetic efficiency and levels of chlorophylls were also measured, indicating that plant stress indicators were identical in all plants independently of the media to which the plants were exposed. PAHs influenced both the soluble Cu fraction and Cu uptake by plants. The amounts of metal accumulated in both roots and stems were significantly higher when the 10(4) microg l(-1) of Cu enriched elutriate was amended with PAHs. Thus, results suggest that PAHs may modify Cu solubility, the Cu sorption by plants and/or the passive penetration of Cu into the root cells. Therefore, the combined effects of different types of pollutants should be taken in consideration when studying the remediation potential of plants, namely in terms of phytoextraction.

Bacterial community response to petroleum contamination and nutrient addition in sediments from a temperate salt marsh.

Microbial communities play an important role in the biodegradation of organic pollutants in sediments, including hydrocarbons. The aim of this study was to evaluate the response of temperate salt marsh microbial communities to petroleum contamination, in terms of community structure, abundance and capacity to degrade hydrocarbons. Sediments un-colonized and colonized (rhizosediments) by Juncus maritimus, Phragmites australis and Triglochin striata were collected in a temperate estuary (Lima, NW Portugal), spiked with petroleum under variable nutritional conditions, and incubated for 15 days. Results showed that plant speciation emerged as the major factor for shaping the rhizosphere community structure, overriding the petroleum influence. Moreover, when exposed to petroleum contamination, the distinct salt marsh microbial communities responded similarly with (i) increased abundance, (ii) changes in structure, and (iii) decreased diversity. Communities, particularly those associated to J. maritimus and P. australis roots displayed a potential to degrade petroleum hydrocarbons, with degradation percentages between 15% and 41%, depending on sediment type and nutritional conditions. In conclusion, distinct salt marsh microbial communities responded similarly to petroleum contamination, but presented different pace, nutritional requirements, and potential for its biodegradation, which should be taken into account when developing bioremediation strategies.

Potential of constructed wetlands microcosms for the removal of veterinary pharmaceuticals from livestock wastewater

The aim of the present work was to evaluate, at microcosm level, the capacity of constructed wetlands (CWs) to remove veterinary pharmaceutical compounds, from wastewater. Results indicated that CWs have potential to mitigate the release of veterinary drugs, namely enrofloxacin (ENR, a fluoroquinolone) and tetracycline (TET, tetracyclines family). Removal efficiencies of 94% and 98% where achieved for TET and ENR, respectively, when treating pigfarm wastewater effluent doped at 100 μg L(-1) drug level, along twelve weeks. Occurrence of adsorption of the drugs to CWs substrate may be the predominant mechanism for ENR, although for TET there are signs that degradation is also occurring.

Influence of surfactants on the Cu phytoremediation potential of a salt marsh plant

To assess the possible effect that surfactants commonly found in the aquatic environment may have on the remediation potential of the salt marsh plant Halimione portulacoides, a non-ionic (Triton X-100) and an anionic (sodium dodecyl sulfate, SDS) surfactants were used. Experiments were carried out in the laboratory, either in hydroponics (sediment elutriate) or in sediment soaked in elutriate, using sediment and water from an estuarine salt marsh (Cávado River, NW Portugal). Groups of H. portulacoides (grown in a greenhouse) were exposed for 6d to media with 0.16mM added Cu(II) in the absence and in the presence of each one of the two selected surfactants, at concentrations lower than the respective micellar critical concentration. Cu was determined in solutions, sediments and in different plant tissues before and after experiments. Plant photosynthetic efficiency did not indicate deletory effects of the exposure to the added pollutants. The non-ionic surfactant Triton X-100 and, to a lesser extent, the anionic surfactant SDS too, favored Cu accumulation in the plant roots but not Cu translocation, indicating that surfactants may favor Cu adsorption to the roots (phytostabilization). On the other hand, both surfactants favored Cu solubility from the sediment. Therefore, the presence of surfactants, which are frequently found in estuarine areas, as a result of urban and industrial effluent discharges, may condition metal distribution in those environments.

Influence of a salt marsh plant (Halimione portulacoides) on the concentrations and potential mobility of metals in sediments

Influence of Halimione portulacoides, commonly found in temperate salt marshes, on sediment metal contents, speciation and potential mobility in case of sediment re-suspension was evaluated. Both colonized and non-colonized sediments were studied for total Cd, Cu, Pb and Zn contents and metal fraction exchangeable to water collected in situ. Sediment elutriates, prepared with water collected from each site, were used to simulate a sediment re-suspension phenomenon. As the characteristics and degree of contamination of sediments may influence system behaviour, salt marshes of two Portuguese estuaries, Cavado (NW coast) and Sado (SW coast), were studied. Cu, Pb and Zn contents higher than ERL (quality guideline, effect range-low) were observed, indicating potential risks for living organisms. Strong Cu-complexing organic ligands, also determined in both water and elutriates, were higher in rhizosediment elutriates, at concentrations similar, or even higher, to those of Cu. Such ligands condition metals speciation in the water column and probably also metal bioavailability. From rhizosediment significant amounts of Cu and Zn were transferred to the aqueous phase, concentrations 2-8 times higher than concentrations present in water. In contrast, elutriates of non-colonized sediment removed metals from water, Cu and Zn levels in elutriates being 2-6 times lower than initial ones. Cd and Pb levels in water and elutriates were not measurable in most cases. Results clearly indicate that metals potential solubility in the rhizosphere of plants was markedly higher than that in the surrounding sediment. The obtained results indicated that H. portulacoides presence (and probably other salt marsh plants) may cause a marked increase in metals concentrations in dissolved phase (pore water or even water column if rhizosediment is re-suspended). As salt marsh plants may be abundant in temperate and subtropical estuaries and costal lagoons, this phenomenon should not be disregard in future studies towards the sustainable management of such environments.

Microbial community dynamics associated with veterinary antibiotics removal in constructed wetlands microcosms

This study aimed to evaluate the response of the microbial community from CWs microcosms tested for the removal of two veterinary antibiotics, enrofloxacin (ENR) and tetracycline (TET), from livestock industry wastewater. Three treatments were tested (control, ENR or TET (100 μg L(-1))) over 12 weeks in microcosms unplanted and planted with Phragmites australis. CWs removal efficiency was relatively stable along time, with removals higher than 98% for ENR and 94% for TET. In addition, CWs were able to reduce wastewater toxicity, independently of antibiotics presence. Despite no significant differences were observed in terms of microbial abundance, bacterial richness or diversity, analysis of similarities (two-way crossed ANOSIM) showed a significant effect of both time and treatments in bacterial community structure. This study points to CWs applicability for veterinary antibiotics removal from livestock wastewaters, showing that CWs microbial communities were able to adapt without significant changes in their diversity or depuration capacity.

Sediment quality in the Douro river estuary based on trace metal contents, macrobenthic community and elutriate sediment toxicity test (ESTT)

The aim of this study was to evaluate the sediment quality in the mesotidal Douro River estuarine environment, in order to identify areas where sediment contamination could cause ecosystem degradation. Samples were obtained in five locations and sediment characterised for grain size, total organic matter, total-recoverable metals (Al, Fe, Cu, Pb, Cr, Ni, Cd, Zn and Mn), as well as acid volatile sulfide (AVS) and simultaneously extracted metals (SEM). In situ effects were evaluated by examining the macrobenthic community structure. An elutriate sediment toxicity test (ESTT) was used to estimate the amount of metals and nutrients that could be exchanged with the water column through resuspension, and its positive or negative effects on the growth of the micro-alga Emiliania huxleyi in a 10 day test. Anthropogenic metal contamination was identified at the north bank of the Douro estuary, with deleterious effects on the macrobenthic community, namely decrease in number of species and diversity. This contamination could possibly also be toxic for water column organisms, in case of resuspension, as shown by the ESTT. Sediments from the salt marsh at the south bank showed an impoverished macrobenthic community and elutriate toxicity, which appeared to be due to anaerobic conditions. This study clearly shows the usefulness of the ESST approach to assess the biological effect of resuspension of estuarine sediments.

Biodegradation of the veterinary antibiotics enrofloxacin and ceftiofur and associated microbial community dynamics

Fluoroquinolones and cephalosporins are two classes of veterinary antibiotics arising as pollutants of emerging concern. In this work, the microbial degradation of two representative antibiotics of both these classes, enrofloxacin (ENR) and ceftiofur (CEF), is reported. Biodegradation of the target antibiotics was investigated by supplementing the culture medium with ENR and CEF, individually and in mixture. Microbial inocula were obtained from rhizosphere sediments of plants derived from experimental constructed wetlands designed for the treatment of livestock wastewaters contaminated with trace amounts of these antibiotics. Selected microbial inocula were acclimated during a period of 5months, where the antibiotics were supplemented every three weeks at the concentration of 1mgL-1, using acetate as a co-substrate. After this period, the acclimated consortia were investigated for their capacity to biodegrade 2 and 3mgL-1 of ENR and CEF. Complete removal of CEF from the inoculated culture medium was always observed within 21days, independently of its concentration or the concomitant presence of ENR. Biodegradation of ENR decreased with the increase in its concentration in the culture medium, with defluorination percentages decreasing from ca. 65 to 4%. Ciprofloxacin and norfloxacin were detected as biodegradation intermediates of ENR in the microbial cultures supplemented with this antibiotic, indicating that defluorination of at least part of ENR in these cultures is not an immediate catabolic step. Abiotic mechanisms showed high influence in the removal of CEF, affecting less ENR degradation. The acclimation process with the target antibiotics led to significant shifts in the structure and diversity of the microbial communities, predominantly selecting microorganisms belonging to the phyla Proteobacteria (e.g. Achromobacter, Variovorax and Stenotrophomonas genera) and Bacteroidetes (e.g. Dysgonomonas, Flavobacterium and Chryseobacterium genera). The results presented in this study indicate that biodegradation can be an important mechanism for the environmental removal of the tested compounds.

Development of autochthonous microbial consortia for enhanced phytoremediation of salt-marsh sediments contaminated with cadmium.

UNLABELLED Microbial assisted phytoremediation is a promising, though yet poorly explored, new remediation technique. The aim of this study was to develop autochthonous microbial consortia resistant to cadmium that could enhance phytoremediation of salt-marsh sediments contaminated with this metal. The microbial consortia were selectively enriched from rhizosediments colonized by Juncus maritimus and Phragmites australis. The obtained consortia presented similar microbial abundance but a fairly different community structure, showing that the microbial community was a function of the sediment from which the consortia were enriched. The effect of the bioaugmentation with the developed consortia on cadmium uptake, and the microbial community structure associated to the different sediments were assessed using a microcosm experiment. Our results showed that the addition of the cadmium resistant microbial consortia increased J. maritimus metal phytostabilization capacity. On the other hand, in P. australis, microbial consortia amendment promoted metal phytoextraction. The addition of the consortia did not alter the bacterial structure present in the sediments at the end of the experiments. This study provides new evidences that the development of autochthonous microbial consortia for enhanced phytoremediation of salt-marsh sediments contaminated with cadmium might be a simple, efficient, and environmental friendly remediation procedure. CAPSULE ABSTRACT Development of autochthonous microbial consortia resistant to cadmium that enhanced phytoremediation by salt-marsh plants, without a long term effect on sediment bacterial diversity.