C. Marisa R. Almeida

A review of plant–pharmaceutical interactions: from uptake and effects in crop plants to phytoremediation in constructed wetlands

Pharmaceuticals are commonly found both in the aquatic and the agricultural environments as a consequence of the human activities and associated discharge of wastewater effluents to the environment. The utilization of treated effluent for crop irrigation, along with land application of manure and biosolids, accelerates the introduction of these compounds into arable lands and crops. Despite the low concentrations of pharmaceuticals usually found, the continuous introduction into the environment from different pathways makes them ‘pseudo-persistent’. Several reviews have been published regarding the potential impact of veterinary and human pharmaceuticals on arable land. However, plant uptake as well as phytotoxicity data are scarcely studied. Simultaneously, phytoremediation as a tool for pharmaceutical removal from soils, sediments and water is starting to be researched, with promising results. This review gives an in-depth overview of the phytotoxicity of pharmaceuticals, their uptake and their removal by plants. The aim of the current work was to map the present knowledge concerning pharmaceutical interactions with plants in terms of uptake and the use of plant-based systems for phytoremediation purposes.

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.

Metal levels in sediments from the Minho estuary salt marsh: a metal clean area?

Total-recoverable metals (Al, Cr, Cu, Fe, Mn, Ni, Pb and Zn) in sediments from Minho estuary salt marsh were determined to evaluate possible increase in anthropogenic contamination by metals and to evaluate the possibility of this area to be considered a pristine area in terms of metals, which can be used as a reference site for other metal-contaminated national and international estuaries/salt marshes. This study revealed that the spatial distribution of metals in the salt marsh sediments was not homogeneous and that two sampling sites (sites 5 and 7) had indications of anthropogenic contamination. However, metal levels in these salt marsh sediments were lower than those observed in the wetlands of the main Portuguese estuaries. Comparison with Portuguese and international reference values used in the evaluation of the ecological quality of sediments, indicated that the sediments can be classified as “clean sediment” and that metal levels were lower or similar (only for Cu and Ni) to the values of ERL, which are the values that define the concentrations ranges that are rarely associated to adverse biologic effects in organisms. In addition, metal levels in the sediments were in chemical forms that were not easily available to organisms, indicating that these sediments probably will not have negative influences in the organisms living in the salt marsh, although no life-form ecological safety tests have been carried out. Therefore, the Minho estuary salt marsh area can probably be considered a pristine area in terms of metals and can be used as a reference for other metal-contaminated estuaries/salt marshes.

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 Phragmites australis for the removal of veterinary pharmaceuticals from aquatic media

The potential of Phragmites australis was evaluated for the removal of three veterinary drugs, enrofloxacin (ENR), ceftiofur (CEF) and tetracycline (TET), from aquatic mediums. Results showed that the plant promoted the removal of 94% and 75% of ENR and TET, respectively, from wastewater. Microbial abundance estimation revealed that microorganisms were not a major participant. Occurrence of drugs adsorption to plant roots was observed in small extension. Therefore, main mechanisms occurring were drug removal by plant uptake and/or degradation. Present results demonstrated the potential of P. australis-planted beds to be used for removal of pharmaceuticals from livestock and slaughterhouse industries wastewater.

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.

The Mammalian "Obesogen" Tributyltin Targets Hepatic Triglyceride Accumulation and the Transcriptional Regulation of Lipid Metabolism in the Liver and Brain of Zebrafish.

Recent findings indicate that different Endocrine Disrupting Chemicals (EDCs) interfere with lipid metabolic pathways in mammals and promote fat accumulation, a previously unknown site of action for these compounds. The antifoulant and environmental pollutant tributyltin (TBT), which causes imposex in gastropod snails, induces an “obesogenic” phenotype in mammals, through the activation of the nuclear receptors retinoid X receptor (RXR) and peroxisome proliferator-activated receptor gamma (PPARγ). In teleosts, the effects of TBT on the lipid metabolism are poorly understood, particularly following exposure to low, environmental concentrations. In this context, the present work shows that exposure of zebrafish to 10 and 50 ng/L of TBT (as Sn) from pre-hatch to 9 months of age alters the body weight, condition factor, hepatosomatic index and hepatic triglycerides in a gender and dose related manner. Furthermore, TBT modulated the transcription of key lipid regulating factors and enzymes involved in adipogenesis, lipogenesis, glucocorticoid metabolism, growth and development in the brain and liver of exposed fish, revealing sexual dimorphic effects in the latter. Overall, the present study shows that the model mammalian obesogen TBT interferes with triglyceride accumulation and the transcriptional regulation of lipid metabolism in zebrafish and indentifies the brain lipogenic transcription profile of fish as a new target of this compound.

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.

Advantages and limitations of the semi-quantitative operation mode of an inductively coupled plasma-mass spectrometer for multi-element analysis of wines

The suitability of the semi-quantitative mode of analysis of a Perkin-Elmer inductively coupled plasma quadrupole-mass spectrometer for multi-element characterization of fortified and table wines was tested. The samples, embracing red and white table, and Port wines, were first submitted to an UV-irradiation pre-treatment. The same samples were analysed, in parallel, by using the quantitative mode of the same equipment, and the results were compared. External calibration with aqueous standards was used in the quantitative mode and calibration with only one aqueous multi-element standard was used in the semi-quantitative mode. Thirty elements, including major, minor and trace elements (Al, Ba, Cd, Co, Cr, Cu, Fe, Ga, Li, Mn, Ni, Pb, Rb, Sr, V, Zn and the rare earth elements (REEs) La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu) were measured by the two modes of analysis. For most elements the semi-quantitative mode provided two to six times higher limits of detection than those obtained by the quantitative mode. The precision of the results were similar for both modes, with relative standards deviation between 0.5 and 5%, in many cases. Comparable values of concentration (differences <20%) were obtained by the two methodologies, except for Cd, Ga, Li and the REEs. A linear least-squares adjustment of the two global sets of results (semi-quantitative mode (y-axis) versus quantitative mode (x-axis)) yielded the equation: y=(0.93±0.01)x+(1±11) with a very high correlation coefficient of 0.9992 (n=40). Although the results obtained by the semi-quantitative mode were, on average, slightly lower than those given by the quantitative mode (slope <1), semi-quantitative analysis can be an important tool for comparison of elemental compositions of wines, including fortified wines. It gives rapid information about the concentration levels of a large number of elements, and presents some economic advantages relatively to the quantitative mode: it is faster and requires fewer reagents.