Santiago Andrade

Distribution of Heavy Metals in Surface Sediments from an Antarctic Marine Ecosystem

The concentrations of lead, cadmium, copper,chromium, iron, manganese and zinc in surfacesediments collected from Potter Cove, in the 25de Mayo Island (King George Island), Antarctica, andits drainage basin, were measured by atomic absorptionspectroscopy. The obtained results were use todetermine the areal and vertical distribution of themetals of in the Cove and potential sources of thesemetals to this environment. The geochemical datasuggest that most of the metals found in Potter Coveconstitute a redistribution of autochthonous materialswithin the ecosystem. Therefore, the metalconcentrations can be considered to be present atnatural background levels in surface sediments.

Heavy metal concentration in mangrove surface sediments from the north-west coast of South America

Mangrove ecosystems are coastal estuarine systems confined to the tropical and subtropical regions. The Estero Salado mangrove located in Guayaquil, Ecuador, has suffered constant disturbances during the past 20 years, due to industrial wastewater release. However, there are no published data for heavy metals present in its sediments and the relationship with anthropogenic disturbance. In the present study, metal concentrations were evaluated in surface sediment samples of the mangrove, showing that B, Cd, Cu, Pb, Se, V, and Zn levels exceeded those declared in international environmental quality standards. Moreover, several metals (Pb, Sn, Cd, Ag, Mo, Zn and Ni) could be linked to the industrial wastewater present in the studied area. In addition, heavy metal levels detected in this mangrove are higher than previous reports on mangrove sediments worldwide, indicating that this mangrove ecosystem is one of the most disrupted on earth.

Changes in bacterial community structure associated with coastal copper enrichment

Marine bacterial communities isolated from the water column, sediment, the rock surface, and the green seaweed Ulva compressa were studied in an intertidal ecosystem. The study area included a coastal zone chronically affected by copper mine waste disposals. Bacterial community composition was analyzed by terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA genes, and multivariate analyses of T-RFLP data sets were used for comparisons. Results showed that diversity and richness indexes were not able to detect differences among compartments. However, comparisons within the same compartment clearly showed that copper enrichment was associated with changes in the composition of the bacterial communities and revealed that the magnitude of the effect depends on the compartment being considered. In this context, communities from sediments appeared as the most affected by copper enrichment. The present study also demonstrated that intertidal bacterial communities were dominated by Gammaproteobacteria, Firmicutes, and Actinobacteria and the changes in these communities were mainly due to changes in their relative abundances.

Changes in epiphytic bacterial communities of intertidal seaweeds modulated by host, temporality, and copper enrichment.

This study reports on the factors involved in regulating the composition and structure of bacterial communities epiphytic on intertidal macroalgae, exploring their temporal variability and the role of copper pollution. Culture-independent, molecular approaches were chosen for this purpose and three host species were used as models: the ephemeral Ulva spp. (Chlorophyceae) and Scytosiphon lomentaria (Phaeophyceae) and the long-living Lessonia nigrescens (Phaeophyceae). The algae were collected from two coastal areas in Northern Chile, where the main contrast was the concentration of copper in the seawater column resulting from copper-mine waste disposals. We found a clear and strong effect in the structure of the bacterial communities associated with the algal species serving as host. The structure of the bacterial communities also varied through time. The effect of copper on the structure of the epiphytic bacterial communities was significant in Ulva spp., but not on L. nigrescens. The use of 16S rRNA gene library analysis to compare bacterial communities in Ulva revealed that they were composed of five phyla and six classes, with approximately 35 bacterial species, dominated by members of Bacteroidetes (Cytophaga-Flavobacteria-Bacteroides) and α-Proteobacteria, in both non-polluted and polluted sites. Less common groups, such as the Verrucomicrobiae, were exclusively found in polluted sites. This work shows that the structure of bacterial communities epiphytic on macroalgae is hierarchically determined by algal species > temporal changes > copper levels.

Novel polymerase chain reaction primers for the specific detection of bacterial copper P-type ATPases gene sequences in environmental isolates and metagenomic DNA.

Aims:  In the last decades, the worldwide increase in copper wastes release by industrial activities like mining has driven environmental metal contents to toxic levels. For this reason, the study of the biological copper‐resistance mechanisms in natural environments is important. Therefore, an appropriate molecular tool for the detection and tracking of copper‐resistance genes was developed.

Abundance and diversity of copper resistance genes cusA and copA in microbial communities in relation to the impact of copper on Chilean marine sediments.

Microorganisms have developed copper-resistance mechanisms in order to survive in contaminated environments. The abundance of the copper-resistance genes cusA and copA, encoding respectively for a Resistance Cell Nodulation protein and for a P-type ATP-ase pump, was assessed in copper and non-copper-impacted Chilean marine sediment cores by the use of molecular tools. We demonstrated that number of copA and cusA genes per bacterial cell was higher in the contaminated sediment, and that copA gene was more abundant than cusA gene in the impacted sediment. The molecular phylogeny of the two copper-resistance genes was studied and reveals an impact of copper on the genetic composition of copA and cusA genes.

Mercury distribution in estuarine environments from Argentina: the detoxification and recovery of salt marshes after 15 years

Total Hg contents from abiotic (surface sediments and suspendedparticulate matter) and biological (crabs, fishes and halophytes)compartments from Bahía Blanca estuary and Mar Chiquita CoastalLagoon, Argentina, have been monitored since the 1980s. At BahíaBlanca estuary, high Hg concentrations were recorded during the early1980s in surface sediments (0.34 ± 0.22 μg/g) andsuspended particulate matter (0.19 ± 0.10 μg/g). Fishspecies, Mustelus schmitti (0.89 ± 0.29 μg/g), Paralichthys brasiliensis (0.85 ± 0.18 μg/g) and Micropogonias furnieri (0.37 ± 0.11 μg/g) also presentedhigh Hg concentrations. The large industrial nucleus located within theestuary has been identified as the main metal source for this environment.Hg contents from the same area during 1996–1998 were significantlylower: surface sediments (0.164 ± 0.023 μg/g), suspendedparticulate matter (0.048 ± 0.0017 μg/g), fish Micropogonias furnieri (0.13 ± 0.02 μg/g) and crab Chasmagnathus granulata (0.334 ± 0.071 μg/g). This trendof environmental detoxification is probably related with (i) thetechnological changes incorporated by the local industry, (ii) a mostadequate management of industrial effluents, and (iii) the removal ofgreat sediment volume by dredging and refill.During the 1980s Mar Chiquita Lagoon Hg concentrations reached 0.08± 0.01 μg/g in surface sediments and 0.09 ±0.025 μg/g in suspended particulate matter, and 0.14 ±0.04 μg/g in the fish Basilichthys bonariensis and 0.22 ±0.08 μg/g in Paralichthys brasiliensis, and 0.08 ±0.01 μg/g in the crab C. granulata, Hg concentrations werelower than at Bahía Blanca. Remote Hg sources for this Coastal Lagoonand atmospheric and stream transport of Hg is proposed as major Hgsources, since no Hg point sources exists nearby. Mercury concentrationsrecorded in the 1996–1998 period were lower than those recorded inthe previous decade: surface sediments (0.019 ± 0.004 μg/g), suspended particulate matter (0.030 ± 0.008 μg/g), halophyte Spartina densiflora (0.013 ± 0.008 μg/g) or crab C. granulata (0.011 ± 0.009 μg/g).Both Hg bioaccumulation and biomagnification processes were verified inBahía Blanca estuary and in Mar Chiquita Coastal Lagoon. This apparentrecovery of both estuarine environments deserves to be carefully analyzed,in order to fully understand the foundations of these processes.

Physiological plasticity of Dictyota kunthii (Phaeophyceae) to copper excess

The brown alga Dictyota kunthii is one of the dominant species in the coastal areas of northern Chile affected by copper enrichment due to accumulated mining wastes. To assess its physiological plasticity in handling copper-mediated oxidative stress, 4-days copper exposure (ca. 100 μg/L) experiments were conducted with individuals from a copper impacted area and compared with the responses of plants from a non-impacted site. Several biochemical parameters were then evaluated and compared between populations. Results showed that individuals from the copper-impacted population normally displayed higher levels of copper content and antioxidant enzymes activity (catalase (CAT), ascorbate peroxidase (AP), dehydroascorbate reductase (DHAR), glutathione peroxidase (GP) and peroxiredoxins (PRX)). After copper exposure, antioxidant enzyme activity increased significantly in plants from the two selected sites. In addition, we found that copper-mediated oxidative stress was associated with a reduction of glutathione reductase (GR) activity. Moreover, metabolic profiling of extracellular metabolites from both populations showed a significant change after plants were exposed to copper excess in comparison with controls, strongly suggesting a copper-induced release of metabolites. The copper-binding capacity of those exudates was determined by anodic stripping voltammetry (ASV) and revealed an increased ligand capacity of the medium with plants exposed to copper excess. Results indicated that D. kunthii, regardless their origin, counteracts copper excess by various mechanisms, including metal accumulation, activation of CAT, AP, DHAR, GP and PRX, and an induced release of Cu binding compounds. Thus, plasticity in copper tolerance in D. kunthii seems constitutive, and the occurrence of a copper-tolerant ecotype seems unlikely.

Eukaryotic picophytoplankton community response to copper enrichment in a metal-perturbed coastal environment

Copper is an essential micronutrient, especially for photosynthetic organisms, but can be toxic at high concentrations. In the past years, coastal waters have been exposed to an increase in copper concentration due to anthropogenic inputs. One well known case is the Chañaral area (Easter South Pacific coast), where a long term coastal copper enrichment event has occurred. That event strongly affected benthic marine diversity, including microbial communities. In this work, microcosm experiments were carried out to address the changes on picophytoplankton community composition of the disturbed area, when challenged to copper additions. Eukaryotic picophytoplankton communities from two areas were analyzed: one in the most copper‐perturbed area and another at the north edge of the perturbed area. Flow cytometry data showed that 25 μg L−1 of copper addition exerted a positive effect in the growth kinetics on part of the eukaryotic picophytoplankton communities, independently of the site. 16S‐plastid terminal restriction fragment length polymorphisms analysis suggested that eukaryotic picophytoplankton display a short and directional response to high copper levels. Members of the Prasinophyceae class, a Coscinodiscophyceae diatom, as well as Phaeocystis, respond in a short time to the environmental disturbance, making them excellent candidates for further studies to evaluate phytoplanktonic species as sentinels for copper disturbances in coastal marine ecosystems.

Ostreococcus tauri Luminescent Reporter Lines as Biosensors for Detecting Pollution From Copper-Mine Tailing Effluents in Coastal Environments

Phytoplankton cells are excellent biosensors for environmental monitoring and toxicity assessments in different natural systems. Green algae, in particular, appear to be more responsive to copper (Cu) disturbances. This is interesting considering that Cu pollution in coastal environments has increased over the last century, with enormous repercussions to marine ecosystems. Unfortunately, no high-throughput method exists for the environmental monitoring of Cu toxicity in seawater. To assess potential uses as biosensors of Cu pollution, high-throughput screening was performed on five luminescence reporter lines constructed in the green algae Ostreococcus tauri RCC745. The reporter line expressing the iron storage ferritin protein fused to luciferase (Fer-Luc) was the most sensitive, responding to Cu concentrations in the µM range. Fer-Luc was also the most sensitive reporter line for detecting toxicity in mining-derived polluted seawater predominantly contaminated by soluble Cu. Nevertheless, the Cyclin-Dependent-Kinase A (CDKA) reporter was most suitable for detecting the toxicity of copper-mine tailing effluents containing other metals (e.g., iron). These results highlight that Ostreococcus biosensors can serve as a reliable, inexpensive, and automated, high-throughput laboratory approach for performing seawater analyses of coastal areas subjected to metal disturbances. When challenged with Cu, Ostreococcus tauri not only evidenced a rapid, transcriptional response for the tested genes, but also showed changes in a broad range of genes, especially as related to the stress response. Overall, the obtained results reinforce that a single biosensor is insufficient when dealing with complex mixtures of toxic compounds in natural environments.