Potential phytoremediation system using macrophyte Limnobium laevigatum to remove in situ Cr from contaminated bottom sediments.

The contamination of aquatic environments with heavy metals is an important issue, and in turn, it is crucial to study remediation techniques that can be applied in situ. In this work, the use of a containment system with macrophytes Limnobium laevigatum is explored in the laboratory to evaluate the remotion of Cr in contaminated sediments. The roots of the plants were placed in contact with the bottom sediment through a containment system. The concentration of Cr in macrophyte and sediment samples exposed to different exposure times (1, 4 and 7 days) was determined by laser-induced breakdown spectroscopy technique. The initial concentration of Cr in the sediment was 112 ± 5 mg/kg and decreased by 65% to the control (p < 0.05) after 24 h of exposure. The removal continued throughout the study time until reaching values of 23 ± 1 mg/kg. In macrophytes, the Cr concentration increased from 20 ± 5 mg/kg to 2066 ± 216 mg/kg after seven days of exposure. The correlation coefficient between Cr concentrations in both matrices was -0.96. Finally, the bioaccumulation factor of Cr in L. laevigatum was 95.22 ± 8.51. Therefore, the system studied could be a potential tool to remedy the bottom sediments of streams and lakes contaminated with heavy metals in situ.

Interaction of Thalassia testudinum Metabolites with Cytochrome P450 Enzymes and Its Effects on Benzo(a)pyrene-Induced Mutagenicity.

The aim of the present work was to evaluate the effects of Thalassia testudinum hydroethanolic extract, its polyphenolic fraction and thalassiolin B on the activity of phase I metabolizing enzymes as well as their antimutagenic effects. Spectrofluorometric techniques were used to evaluate the effect of tested products on rat and human CYP1A and CYP2B activity. The antimutagenic effect of tested products was evaluated in benzo[a]pyrene (BP)-induced mutagenicity assay by an Ames test. Finally, the antimutagenic effect of Thalassia testudinum (100 mg/kg) was assessed in BP-induced mutagenesis in mice. The tested products significantly (p < 0.05) inhibit rat CYP1A1 activity, acting as mixed-type inhibitors of rat CYP1A1 (Ki = 54.16 ± 9.09 µg/mL, 5.96 ± 1.55 µg/mL and 3.05 ± 0.89 µg/mL, respectively). Inhibition of human CYP1A1 was also observed (Ki = 197.1 ± 63.40 µg/mL and 203.10 ± 17.29 µg/mL for the polyphenolic fraction and for thalassiolin B, respectively). In addition, the evaluated products significantly inhibit (p < 0.05) BP-induced mutagenicity in vitro. Furthermore, oral doses of Thalassia testudinum (100 mg/kg) significantly reduced (p < 0.05) the BP-induced micronuclei and oxidative damage, together with an increase of reduced glutathione, in mice. In summary, Thalassia testudinum metabolites exhibit antigenotoxic activity mediated, at least, by the inhibition of CYP1A1-mediated BP biotransformation, arresting the oxidative and mutagenic damage. Thus, the metabolites of T. testudinum may represent a potential source of chemopreventive compounds for the adjuvant therapy of cancer.

Growth of the aquatic macrophyte Ricciocarpos natans (L.) Corda in different temperatures and in distinct concentrations of aluminum and manganese.

One of the consequences of global mining is the exposure of metals into the environment, caused by the rupture of tailings dams. Excess of metals, such as aluminum (Al) and manganese (Mn) can cause serious damage to fauna and flora. The presence of these metals, associated with the temperature increase that occurs nowadays can potentially increase biochemical and metabolic rates in plant tissues and may affect growth. Therefore, the objective of this work was to evaluate the toxicity of the metals Al and Mn into the biomass' growth of the macrophyte Ricciocarpos natans, under two temperatures (25 and 27 °C). R. natans individuals (n = 10 ± 0.5 cm wide) were exposed during 30 days to Al (1.5; 2.5 and 5.0 mg L-1) and Mn (0.7; 1.5 and 3.0 mg L-1) at temperatures and photoperiod-controlled germination chambers. Fresh macrophyte masses were determined gravimetrically to determine the kinetic growth using a logistic model. With that, it was noticed that the presence of Al interfered negatively in the increase of the R. natans biomass, mainly in the highest concentrations and at 27 °C. Mn, on the other hand, affected the increase in biomass, mainly in the highest concentration. As a result, the growth coefficients (µ) changed, being up to 4 times lower in the Al bioassays and up to 2 times higher than the control in the Mn bioassays. However, the dry R. natans biomass individuals that were exposed to the treatments was reduced when compared to the control, except for the lower concentration of Mn. These results contribute to the understanding of the environmental changes that can occur due to metals contained in mining tailings in aquatic ecosystems and the influence of global warming on the metabolic processes of the growth of aquatic macrophytes.

Effect of the exogenous anthocyanin extract on key metabolic pathways and antioxidant status of Brazilian elodea (Egeria densa (Planch.) Casp.) exposed to cadmium and manganese.

Present study deals with the effect of 24 h pre-incubation with exogenous anthocyanins (ANTH), extracted from red cabbage leaves, on key metabolic processes (photosynthesis and respiration) and pro-/antioxidant balance in the aquatic macrophyte Egeria densa (Planch.) Casp., Hydrocharitaceae family, treated with Cd and Mn (in sulfate form) at a concentration of 100 µmol. After five days of metal treatments, Cd was accumulated and the damage caused to metabolic processes was stronger than Mn. In Cd-treated leaves, the protein level, chlorophyll concentration and maximal photochemical efficiency of PS II decreased twofold, and net-photosynthesis was significantly inhibited, whereas lipid peroxidation and H2O2 production increased. In turn, protective responses developed, including an increase in the total soluble thiols, alternative respiratory pathway capacity and the activity of superoxide dismutase and peroxidases. Pre-incubation in the ANTH-enriched extract caused an increase in foliar ANTH content, enhanced Cd and reduced Mn uptake into the tissue. A decrease in the level of oxidative reactions, an increase in the protein and chlorophyll concentration compared to the control values and a partial improvement of the photosynthetic parameters confirmed the ability of ANTH to reduce Cd-induced damage effects and to mitigate ROS-driven stress reactions. Stimulation of catalase and ascorbate peroxidase activity, an alternative respiration capacity and non-enzymatic antioxidant (carotenoids, ascorbate and proline) synthesis by ANTH were also revealed. These data suggest that ANTH-enriched extract from red cabbage leaves has a protective action against metal toxicity in Egeria plants.

Phytoextraction of Pb, Cr, Ni, and Zn using the aquatic plant Limnobium laevigatum and its potential use in the treatment of wastewater.

In order to study the bioaccumulation of Pb, Cr, Ni, and Zn and the stress response, the floating aquatic plant Limnobium laevigatum was exposed to increasing concentrations of a mixture of these metals for 28 days, and its potential use in the treatment of wastewater was evaluated. The metal concentrations of the treatment 1 (T1) were Pb 1 µg L-1, Cr 4 µg L-1, Ni 25 µg L-1, and Zn 30 µg L-1; of treatment 2 (T2) were Pb 70 µg L-1, Cr 70 µg L-1, Ni 70 µg L-1, and Zn 70 µg L-1; and of treatment 3 (T3) were Pb 1000 µg L-1, Cr 1000 µg L-1, Ni 500 µg L-1, and Zn 100 µg L-1, and there was also a control group (without added metal). The accumulation of Pb, Cr, Ni, and Zn in roots was higher than in leaves of L. laevigatum, and the bioconcentration factor revealed that the concentrations of Ni and Zn in the leaf and root exceeded by over a thousand times the concentrations of those in the culture medium (2000 in leaf and 6800 in root for Ni; 3300 in leaf and 11,500 in root for Zn). Thus, this species can be considered as a hyperaccumulator of these metals. In general, the changes observed in the morphological and physiological parameters and the formation of products of lipid peroxidation of membranes during the exposure to moderate concentrations (T2) of the mixture of metals did not cause harmful effects to the survival of the species within the first 14 days of exposure. Taking into account the accumulation capacity and tolerance to heavy metals, L. laevigatum is suitable for phytoremediation in aquatic environments contaminated with moderated concentrations of Cr, Ni, Pb, and Zn in the early stages of exposure.

Effect of operating temperature on anaerobic digestion of the Brazilian waterweed Egeria densa and its microbial community.

To develop an effective treatment for the globally invasive Brazilian waterweed Egeria densa, anaerobic digestion was observed at 37 °C, 55 °C, and 65 °C. The average methane production rate at 55 °C was 220 mL L-1 day-1, which was two-fold that at 37 °C and 65 °C. Volatile fatty acid accumulation was detected under thermophilic conditions; however, although there was methane production, the system did not shutdown. The microbial communities differed between mesophilic (37 °C) and thermophilic (55 °C and 65 °C) conditions. A bacterial community consisting of the phyla Bacteroidetes (43%), Firmicutes (37%), Proteobacteria (9%), Synergistetes (5%), Spirochaetes (1%), and unclassified bacteria (5%) were detected under mesophilic condition. In contrast, the phylum Firmicutes was dominant under thermophilic conditions. In the archaeal community, Methanosaeta concilii (40%), Methanolinea sp. (17%), and unclassified euryarchaeota (43%) were detected under mesophilic condition. Methanosarcina thermophila (87% at 55 °C, 54% at 65 °C) and Methanothermobacter thermautotrophicus (13% at 55 °C, 46% at 65 °C) were detected under thermophilic conditions. At both 37 °C and 55 °C, acetoclastic methanogenesis likely occurred because of the lower abundance of hydrogenotrophic methanogens. At 65 °C, the growth of the acetoclastic methanogen Methanosarcina thermophila was limited by the high temperature, therefore, acetate oxidation and hydrogenotrophic methanogenesis may have occurred.

Seasonal behavior of Thalassia testudinum (Hydrocharitaceae) metabolites.

The marine angiosperm Thalassia testudinum, commonly known as turtle grass, is a dominant seagrass that grows in the Caribbean Sea shelf associated to Syringodium filiforme. The hydroalcoholic extract of T. testudinum is rich in polyphenols; the most abundant metabolite in this extract is thalassiolin B, a glycosilated flavonoid with skin damage repairing properties, and antioxidant capacity among others. The present study aimed at generating information about the seasonal behavior of secondary metabolites, as well as to study the antioxidant capacity of the T. testudinum leaves extract, collected monthly during 2012 from the Northeast coastline of Havana, Cuba. For this study, spectrophotometric methods were used to determine the concentrations of polyphenols, flavonoids, anthocyanins, soluble carbohydrates and proteins, chlorophylls a and b, and antioxidant activity of the extracts. In general, results demonstrated seasonal variations of the analyzed parameters. Extracts prepared from the vegetal material collected in October and November showed the highest values of polyphenols (58.81 ± 1.53 and 52.39 ± 0.63 mg/g bs, respectivally) and flavonoids (44.12 ± 1.30 and 51.30 ± 0.67 mg/gdw, respectively). On the contrary, the lowest values of polyphenols were found in extracts of leaves collected in July and August (15.51 ± 0.84 and 13.86 ± 0.48 mg/g,respectively). In accordance with these results, the lower value of Inhibitory Concentration (IC50) was obtained to get a 50 % of maximal effect on free radical scavenging activity with the extracts prepared from leaves collected in October and November, and less significant IC50 was obtained from the extract prepared from leaves collected in August (5.63 mg/mL). A negative correlation (r= -0.694) was observed in this study between the content of polyphenols and the IC50 necessary to get the half of its antioxidant maximal effect. The high correspondence between the maximum values of polyphenols, flavonoids, carbohydrates and proteins in October and November, revealed a close relationship between these metabolites found in the extract of T. testudinum. Our hypothesis about the annual variation in the concentration of these metabolites was validated; and these results will support the correct harvesting of T. testudinum leaves for biotechnology and industrial purposes.

Time dependent uptake, bioaccumulation and biotransformation of cell free crude extract microcystins from Lake Amatitlán, Guatemala by Ceratophyllum demersum, Egeria densa and Hydrilla verticillata.

Recent studies evidence that macrophytes can uptake and bioaccumulate microcystins (MC) from contaminated environments, suggesting their use in phytoremediation. In the present study Ceratophyllum demersum, Egeria densa and Hydrilla verticillata were exposed to cell free crude extracts (CE) containing three MC congeners MC-LR, MC-RR and MC-YR at a total MC concentration of 104.4 ± 7.6 µg/L from Lake Amatitlán, Guatemala. Time dependent total glutathione (tGSH), glutathione disulfide (GSSG), disappearance of MC from exposure medium and macrophyte uptake as well as calculated uptake and biotransformation rates and bioconcentration factors (BCF) were monitored after 1, 4, 8 hours (h) and 1, 3, 7 and 14 days (d). Results showed that tGSH concentrations in all exposed macrophytes were enhanced by CE. Disappearance of 62.1 ± 13, 40.8 ± 3.1 and 37.8 ± 3.5 µg/L total MCs from exposure mediums with E. densa, H. verticillata and C. demersum were observed after 1 h. Followed by the total elimination of MCs in exposure medium from H. verticillata after 14 d. Highest MC bioaccumulation capacity (BCF), was observed in E. densa followed by C. demersum and H. verticillata. The here presented results imply the strong MC phytoremediation potential of the evaluated macrophytes.

Eutrophication threatens Caribbean seagrasses - An example from Curaçao and Bonaire.

Seagrass beds are globally declining due to human activities in coastal areas. We here aimed to identify threats from eutrophication to the valuable seagrass beds of Curaçao and Bonaire in the Caribbean, which function as nursery habitats for commercial fish species. We documented surface- and porewater nutrient concentrations, and seagrass nutrient concentrations in 6 bays varying in nutrient loads. Water measurements only provided a momentary snapshot, due to timing, tidal stage, etc., but Thalassia testudinum nutrient concentrations indicated long-term nutrient loads. Nutrient levels in most bays did not raise any concern, but high leaf % P values of Thalassia in Piscadera Bay (∼0.31%) and Spanish Water Bay (∼0.21%) showed that seagrasses may be threatened by eutrophication, due to emergency overflow of waste water and coastal housing. We thus showed that seagrasses may be threatened and measures should be taken to prevent loss of these important nursery areas due to eutrophication.

Direct contribution of the seagrass Thalassia testudinum to lime mud production.

Seagrass beds contribute to oceanic carbonate lime mud production by providing a habitat for a wide variety of calcifying organisms and acting as efficient sediment traps. Here we provide evidence for the direct implication of Thalassia testudinum in the precipitation of aragonite needles. The crystals are located internally in the cell walls, and as external deposits on the blade, and are similar in size and shape to the aragonite needles reported for modern tropical carbonate factories. Seagrass calcification is a biological, light-enhanced process controlled by the leaf, and estimates of seagrass annual carbonate production in a Caribbean reef lagoon are as significant as values reported for Halimeda incrassata. Thus, we conclude that seagrass calcification is another biological source for the aragonite lime mud deposits found in tropical banks, and that tropical seagrass habitats may play a more important role in the oceanic carbon cycle than previously considered.

Changes in trace metals in Thalassia testudinum after hurricane impacts.

Major hurricanes Emily and Wilma hit the Mexican Caribbean in 2005. Changes in trace metals in the seagrass Thalassia testudinum prior to (May 2004, 2005) and following passage of these hurricanes (May, June 2006) were determined at four locations along a ≈ 130 km long stretch of coast. Before the hurricanes, essential metals were likely limiting and concentrations of potentially toxic Pb were high in a contaminated lagoon (27.5 µg g(-1)) and near submarine springs (6.10 µg g(-1)); the likely sources were inland sewage disposal or excessive boat traffic. After the hurricanes, Pb decreased to 2.0 µg g(-1) in the contaminated lagoon probably through flushing. At the northern sites, essential Fe increased >2-fold (from 26.8 to 68.3 µg g(-1) on average), possibly from remobilization of anoxic sediments or upwelling of deep seawater during Wilma. Thus, hurricanes can be beneficial to seagrass beds in flushing toxic metals and replenishing essential elements.