Unravelling the metal uptake process in mosses: Comparison of aquatic and terrestrial species as air pollution biomonitors.

Transplanted mosses have been widely shown to be excellent tools for biomonitoring air pollution; however, it is not clear how the functional groups present on their surfaces affect the uptake of metal cations. In the present study, we examined differences in trace metal accumulation in two terrestrial and one aquatic moss species, and investigated whether the differences depended on their physico-chemical characteristics. In the laboratory, we determined C, N and H contents in their tissues and obtained the ATR-FTIR spectra (to identify the presence of functional groups). We also conducted surface acid-base titrations and metal adsorption assays with Cd, Cu and Pb. In the field, we exposed transplants of each species near different air-polluting industries, and determined the mosses enrichment of Al, Cd, Co, Cr, Cu, Fe, Ni, Pb and V. Laboratory results demonstrated higher metal uptake capacity in the terrestrial mosses Sphagnum palustre and Pseudoscleropodium purum, compared to that in the aquatic moss Fontinalis antipyretica, which can be attributed to a greater abundance of acidic functional groups (i.e. negatively charged binding sites) on the surface of the terrestrial mosses. The affinity of moss for certain elements depends on the abundance and nature of surface functional groups. Accordingly, the metal concentrations generally reached higher levels in S. palustre transplants compared to the other species, except for the uptake of Hg, which was higher in F. antipyretica. However, the findings also suggest an interaction between the type of environment (terrestrial or aquatic) and the moss characteristics that may influence the abovementioned trend. Thus, irrespective of the physico-chemical characteristics, metal uptake varied depending on the environment of origin of the mosses "i.e. atmospheric or aquatic". In other words, the findings suggest that species that accumulate more metals in terrestrial environments will accumulate lower amounts of metals in aquatic environments and vice versa.

Global decrease in heavy metal concentrations in brown algae in the last 90 years.

In the current scenario of global change, heavy metal pollution is of major concern because of its associated toxic effects and the persistence of these pollutants in the environment. This study is the first to evaluate the changes in heavy metal concentrations worldwide in brown algae over the last 90 years (>15,700 data across the globe reported from 1933 to 2020). The study findings revealed significant decreases in the concentrations of Cd, Co, Cr, Cu, Fe, Hg, Mn, Pb and Zn of around 60-84% (ca. 2% annual) in brown algae tissues. The decreases were consistent across the different families considered (Dictyotaceae, Fucaceae, Laminariaceae, Sargassaceae and Others), and began between 1970 and 1990. In addition, strong relationships between these trends and pH, SST and heat content were detected. Although the observed metal declines could be partially explained by these strong correlations, or by adaptions in the algae, other evidences suggest an actual reduction in metal concentrations in oceans because of the implementation of environmental policies. In any case, this study shows a reduction in metal concentrations in brown algae over the last 50 years, which is important in itself, as brown algae form the basis of many marine food webs and are therefore potential distributors of pollutants.

Using MixSIAR to quantify mixed contributions of primary producers from amino acid δ15N of marine consumers.

Estimations of the trophic position and the food web nitrogen baseline from compound-specific isotope analysis of individual amino acids (CSIA-AA) are challenged when the diet of consumer organisms relies on different proportions of vascular and non-vascular primary producers. Here we propose a method to infer such proportions using mixing models and the δ15N CSIA-AA values from marine herbivores. Combining published and new data, we first characterized CSIA-AA values in phytoplankton, macroalgae and vascular plants, and determined their characteristic ß values (i.e. the isotopic difference between trophic and source AA). Then, we applied MixSIAR Bayesian isotope mixing models to investigate the transfer of these isotopic signals to marine herbivores (molluscs, green turtles, zooplankton and fish), and their utility to quantify autotrophic sources. We demonstrated that primary producer groups have distinct δ15NAA fingerprints that can be tracked into their primary consumers, thus offering a rapid solution to quantify resource utilization and estimate ßmix values in mixed-sourced environments.

Analysis of intra-thallus and temporal variability of trace elements and nitrogen in Fucus vesiculosus: Sampling protocol optimization for biomonitoring.

To advance the methodological standardization of the biomonitoring technique using macroalgae, we comprehensively characterized the intra-thallus and temporal patterns of variation in concentrations of a wide set of elements (Al, As, Cd, Co, Cr, Cu, Fe, Hg, Ni, Pb, Zn, N) and δ15N signal in 6 consecutive Fucus vesiculosus apical dichotomous sections collected monthly over a four-year period (2015-2019) at 3 sites on the NW coast of Spain. The concentrations of Al, Co, Fe, Ni, Pb and Zn increased significantly from the youngest to the oldest dichotomies regardless of the sampling time and collection site; As, Cd, N and δ15N showed the opposite trend. Time series analysis revealed a significant and consistent seasonal variation of As, Cd, Co, Cu, Fe, Hg, Ni, Zn, N and δ15N concentrations, with maximum values in winter and minimum values in summer. We discussed the possible mechanisms driving these two sources of variation, and proposed an efficient and effective sampling strategy to minimize their impact in the results of biomonitoring studies, in which the part of the algal thallus selected for chemical analysis and the sampling frequency were carefully considered. This protocol will improve the conclusions and comparability of biomonitoring data from coastal environments.

Optimal number of Fucus vesiculosus samples to differentiate between sites affected by distinct levels of heavy metal contamination.

The presence of trace elements in marine habitats is a serious environmental problem which increasingly affects ecosystem and human health. The use of macroalgae as contamination biomonitors represents a valuable alternative approach to traditional physicochemical methods. The present study was carried out to determine the optimal number of samples of Fucus vesiculosus needed to detect statistically significant differences in the mean concentrations of Al, As, Cd, Co Cr, Cu, Fe, Hg, Ni, Pb, Zn, N and δ15N between two sampling sites affected by different levels of contamination. For this purpose, we plotted the density distributions of the concentrations of the different elements and examined the local variability at three sites. For sites with mean concentrations differing by more than 30 %, 20 samples were sufficient to detect significant differences for all of the elements, except Cr. The proposed methodology could be used in other studies in the absence of specific research on each species and region.

Sampling optimization for biomonitoring metal contamination with marine macroalgae.

The aim of the present study was to optimize the protocol for sampling marine macroalgae to be used to biomonitor heavy metal contamination in marine ecosystems. For this purpose, we collected 50 subsamples of the brown seaweed Fucus vesiculosus at random in each of three sampling sites (SS) and determined the concentrations of Al, As, Cd, Co, Cr, Cu, Fe, Hg, N, Ni, Pb, Zn and δ15N. We used semivariograms to explore the possible existence of spatial structure in the concentrations of the elements. Spatial structure was observed in 88% of the semivariograms studied, with element concentrations varying longitudinally and transversally along the SS. Using randomization techniques, we estimated that in each SS, a minimum of 30 evenly distributed subsamples should be collected within three bands parallel to the coastline (and also at different heights on the rocks if necessary), and analyzed in a single composite sample representative of the intra-SS variability.

Application of macroalgae analysis to assess the natural variability in selected pollution concentrations (N and Hg), and to detect sources of it in coastal environments.

Here we present a new method in which algae are used to detect sources of heavy metal and organic pollution in coastal areas. The procedure involves characterization of the natural range of concentrations of the elements in areas not affected by local sources of pollution and subsequent comparison of concentrations in the study site to these natural range levels. To develop the method, the concentrations of various elements were determined in specimens of the macroalgae Fucus vesiculosus collected at >150 sampling sites along the shoreline. The natural variability in the element concentrations in these zones was established by determining the differences in the tissue concentrations of the elements between pairs of samples separated by different distances. The method was then tested in the surroundings of possible sources of nitrogen and was found to be a tool for detecting sources of small scale nitrogen contamination and for monitoring and evaluating water quality.

Biomonitoring coastal environments with transplanted macroalgae: A methodological review.

The use of macroalgae transplants is a recent technique used in pollution biomonitoring studies in marine ecosystems. Only 60 articles published between 1978 and 2017 reported the use of this environmental tool for the active biomonitoring of inorganic pollutants and nutrients worldwide. In this review paper, we evaluated studies on this topic in relation to the development of methodological aspects of the technique and the degree of standardization of the protocols used. On the basis of findings of this review, we conclude that the technique is not yet standardized and that uniformisation of protocols is required to enable comparison of the results of different studies. We propose a new protocol for applying the technique, in which each suggestion has been carefully and rigorously compared with the relevant findings reported in the available literature.

Evaluation of the use of moss transplants (Pseudoscleropodium purum) for biomonitoring different forms of air pollutant nitrogen compounds.

We investigated whether three different types of moss transplants (devitalized moss bags with and without cover and auto-irrigated moss transplants) are suitable for use as biomonitors of the deposition of oxidised and/or reduced forms of N. For this purpose, we determined whether the concentration of atmospheric NO2 was related to the % N, δ(15)N and the activity of the enzyme biomarkers phosphomonoesterase (PME) and nitrate reductase (NR) in the tissues of moss transplants. We exposed the transplants in 5 different environments of Galicia (NW Spain) and Cataluña (NE Spain): industrial environments, urban and periurban environments, the surroundings of a cattle farm and in a monitoring site included in the sampling network of the European Monitoring Programme. The results showed that the moss in the auto-irrigated transplants was able of incorporating the N in its tissues because it was metabolically active, whereas in devitalized moss bags transplants, moss simply intercepts physically the N compounds that reached it in particulate or gaseous form. In addition, this devitalization could limit the capacity of moss to capture gaseous compounds (i.e. reduced N) and to reduce the oxidised compounds that reach the specimens. These findings indicate that devitalized moss transplants cannot be used to monitor either oxidised or reduced N compounds, whereas transplants of metabolically active moss can be used for this purpose. Finally, the NR and PME biomarkers should be used with caution because of the high variability in their activities and the limits of quantification should be evaluated in each case.

Study of temporal trends in mercury concentrations in the primary flight feathers of Strix aluco.

Temporal trends in Hg concentrations were determined in the primary flight feathers of 146 specimens of Strix aluco which had died in various Wildlife Recovery Centres in Galicia (NW Spain) between 1997 and 2014. The aim of the study was to determine whether standardization of a primary flight feather (or feathers) in this species is essential for identifying temporal trends in Hg concentrations. For this purpose, we had to first standardize the feather(s) analyzed to enable comparison of the levels of Hg detected in different feathers. The results show a high degree of both inter and intra-individual variability but despite that, it was possible to identify P5 as the most representative feather taking into account the amount of metal excreted in each feather and the intra-individual variability: its median was 133ng, which represents 15% (from 7% to 15%) of the total Hg present in all the primary feathers. However, this "standard feather" did not reveal any temporal trend in Hg concentrations for the study period. This lack of trend was found irrespective of the feather considered and it is expected that detection of any existing trend would also not depend on the feather considered. We conclude that use of any particular feather is not essential for identifying temporal trends in Hg concentrations, because the pattern will be identified regardless of the feather selected.