Fontinalis Antipyretica Research Articles

Relationship between potentially toxic elements and macrophyte communities in the Sava river

Freshwater ecosystems are at significant risk of contamination by potentially toxic elements (PTEs) due to their high inherent toxicity, their persistence in the environment and their tendency to bioaccumulate in sediments and living organisms. We investigated aquatic macrophyte communities and the concentrations of As, Cu, Cd, Cr, Pb, Zn, Ni and Fe in water and sediment samples to identify a pollution pattern along the Sava River and to investigate the potential impact of these PTEs on the diversity and structure of macrophyte communities. The study, which covered 945 km of the Sava River, showed a downstream increase in sediment concentrations of the analyzed elements. Both species richness and alpha diversity of macrophyte communities also generally increase downstream. Ordinary and partial Mantel tests indicate that macrophyte communities are significantly correlated with sediment chemistry, but only weakly correlated with water chemistry. In the lowland regions (downstream), beta diversity decreases successively, which can be attributed to an increasing similarity of environmental conditions at downstream sites. Species richness is relatively low at sites with low concentrations of Cr, Cd, Fe, and Cu in the sediment. However, species richness increases to a certain extent with increasing element concentrations; as element concentrations increase further, species richness decreases, probably as a result of increased toxicity. Some species that are generally more tolerant to high concentrations of PTEs are: Ceratophyllum demersum, Iris pseudacorus, Najas marina, Butomus umbellatus, Vallisneria spiralis, Potamogeton gramineus and Bolboschoenus maritimus maritimus. Potamogeton perfoliatus and the moss species Cinclidotus fontinaloides and Fontinalis antipyretica have narrow ecological amplitudes in relation to the concentrations of PTEs in the sediment.

Influence of storage method on the content of photosynthetic pigments of the aquatic moss Fontinalis antipyretica

Aquatic bryophytes are frequently used as bioindicators of water quality in rivers. Poor water quality increases physiological stress in moss, and stress levels can be estimated by measuring the concentrations of photosynthetic pigments and by calculating pigmentary indices. However, studies collecting many samples in a short time may need to store them until analysis. In the meantime, the pigments may suffer some degradation. Given the lack of studies on this problem for aquatic bryophytes, this one investigates the effect of three storage methods (refrigerated at 6 °C, frozen at − 20 °C and dried at room temperature) on the concentrations of chlorophyll a, chlorophyll b, pheophytin a and the following ratios: chlorophyll a/chlorophyll b, D430/D410, D665/D665a, D430/D665 and D480/D665 (where Dx is the absorbance at x nm) in the aquatic moss Fontinalis antipyretica. The results showed that refrigeration was the most suitable method, freezing might be suitable for some parameters, and drying was inadequate for this purpose. We recommend that all studies on photosynthetic pigments detail the time elapsed from collection to sample analysis, as well as the storage method used.

What potential do mosses have as biomonitors of POPs? A comparative study of hexachlorocyclohexane sorption

Persistent organic pollutants (POPs) pose a significant global threat to human health and the environment, and require continuous monitoring due to their ability to migrate long distances. Active biomonitoring using cloned mosses is an inexpensive but underexplored method to assess POPs, mainly due to the poor understanding of the loading mechanisms of these pollutants in mosses. In this work, Fontinalis antipyretica (aquatic moss) and Sphagnum palustre (terrestrial moss) were evaluated as potential biomonitors of hexachlorocyclohexanes (HCHs: α-, β-, γ-, δ-HCH), crucial POPs. Moss clones, grown in photobioreactors and subsequently oven-dried, were used. Their lipid composition and distribution were characterized through molecular and histochemical studies. Adsorption experiments were carried out in the aqueous phase using the repeated additions method and in the gas phase using an active air sampling technique based on solid-phase extraction, a pioneering approach in moss research. F. antipyretica exhibited greater lipid content in the walls of most cells and higher adsorption capacity for all HCH isomers in both gaseous and liquid environments. These findings highlight the need for further investigation of POP loading mechanisms in mosses and open the door to explore other species based on their lipid content.

A method for studying the influence of Mn oxyhydroxides on the trace element content of aquatic bryophytes

The main objective of this study was to develop and test a method of separating externally deposited Mn oxyhydroxides and co-precipitated elements from samples of aquatic moss (the moss Fontinalis antipyretica). The method, which uses 0.1 M hydroxylamine to dissolve the oxyhydroxides, was tested with samples collected in rivers with slightly acidic, well‑oxygenated waters, where high rates of Mn precipitation occur. The method was effective (it extracted up to 84 % of the Mn) and selective (Fe oxyhydroxides were not extracted). The elements Ba, Cd, Zn and Ni were associated with the Mn oxyhydroxides, while Al, As, Cr, Cu, Fe, Hg and Pb were not. Deposition of Mn therefore increased the concentration of some elements in the moss samples. However, as Mn precipitation depends on Eh and pH, which are independent of the concentrations of the elements in water, the relationship between water and moss element concentrations is not clear (i.e. the data are noisy). This is a problem in biomonitoring studies, which assume a close relationship between element concentrations in moss and water. The value of the proposed extraction method is that it can be used to correct the effect of Mn deposition. We present an example of this correction applied to the Cd concentrations in the test data. We found that the noise introduced by the Mn, including age-related effects (observed by comparing concentrations in 0–2.5 and 2.2–5.0 cm sections from the shoot apex), can be reduced. Additionally, the correction revealed recent increases in Cd concentrations in one site that were not observed in the uncorrected data. Another finding of interest was the low content of total Mn and different extractability (of most elements) observed in moss samples collected in alkaline waters. Finally, we discuss how future studies designed for different environmental scenarios can benefit from application of the proposed method.

Particulate matter from water moss of a large Siberian river: Morphometric, mineral, elemental and radionuclide composition

Particulate matter trapped by tufts of water moss Fontinalis antipyretica inhabiting fast flowing waters of the Yenisei River (Siberia, Russia) contaminated with artificial radionuclides has been studied as a potential monitor of radioactive releases to the river. Particulate matter, which was removed from wet tufts of water moss of the Yenisei by rinsing them in water, constituted at least 38% of bulk dry weight of the moss biomass sample and was similar in the contents of chemical elements, minerals, organic matter, and artificial radionuclides to bottom sediments of the Yenisei. Considerable bulk percentages of artificial radionuclides in the sample of water moss, 77% of 137Cs, 44% of 60Co, 41% of 152Eu, 55% of 154Eu, 66% of 241Am, and 34–36% of plutonium were associated with extracellular particles. The comparative study and correlation analysis suggested that 137Cs was mainly associated with mineral particles trapped by moss and that organic matter was responsible for binding plutonium in samples of water moss. Consequently, analysis of extracellular particles of water moss can provide data on contents and speciation of radionuclides transported by water current. Although a considerably high proportion of particulate matter had been washed out from tufts of water moss, some extracellular mineral particulate material and a large number of epiphytic diatoms remained attached to leaves of water moss. Our study proves that particulate matter trapped by water moss can be used as an informative monitor to trace radioactive pollutants transported by water current in running waters deficient in bottom sediments and potential biomonitors.

Assemblages of Trichoptera larvae on water moss in the middle reaches of the Yenisei River (Siberia, Russia).

This study provides first data on species composition of trichopterans inhabiting water moss, Fontinalis antipyretica Hedw., in the middle reaches of the Yenisei River. Analysis of Trichoptera larvae collected from samples of water moss in tail waters of the Krasnoyarsk Hydroelectric Plant in 2008-2021 revealed nine species of the order, which belonged to four families. Among them, six species belonging to the Limnephilidae family have not been mentioned in previous studies of the Yenisei invertebrates. Images of species are provided. One species, Apatania crymophyla McLachlan, was the most dense in all samples of water moss. The densities of A. crymophyla in samples of water moss in 2021 ranged within 1.2-7.0 thousand individuals / kg f.w. Other representatives of the order occurred in samples of water moss sporadically or consistently but in very low densities, which did not allow reliable quantification of their abundances. Consequently, numerous large clumps of water moss in the fast-flowing waters of the Yenisei favor an abundance of Trichoptera larvae, providing these insects refuge, food, and material for their cases. However, the reasons for the predominance of one species, A. crymophyla, are to be elucidated in further studies. The present study suggests that the contributions of Trichoptera and the total invertebrate community inhabiting water moss to biomass and energy budgets in the Yenisei have been usually underestimated before now. Hence, seasonal trends of species composition and abundance of invertebrates associated with water moss in the Yenisei River, including comparison with the entire zoobenthic community, are to be studied in the future.

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.

Influence of the storage procedure on the trace element content measured in the aquatic moss Fontinalis antipyretica Hedw

Aquatic bryophytes have been used as pollution biomonitors for decades. Despite this, sample collection and preparation methods have not been standardized, which makes it difficult to compare the results of different studies. Most times, the samples have to be stored before processing, for example, when many of them are collected in a short time, as occurs in extensive pollution studies. Storage must be done in a way that does not change the pollutant concentrations in the samples. We studied whether the concentrations of Al, As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, and Zn in the aquatic moss Fontinalis antipyretica were affected by three storage procedures: dry at room temperature, fresh (in refrigerator at 6°C), and frozen at -20°C. In addition, we evaluated whether the subsequent washing of the samples affected the concentrations of these elements differently depending on the storage method. Our results showed that the three methods were, in general, adequate since the concentrations did not change, and we did not observe differences between washed and unwashed samples either. Since the simplest method is refrigeration, we concluded that this is the best of them. However, the concentrations of Hg increased steadily over time in the fresh material, probably because of redistribution after volatilization from the basal parts of the mosses. We believe that the respiration of the plants lowered the concentrations of oxygen inside the hermetically sealed bags containing the samples, thus promoting the reduction of the Hg and its posterior volatilization and redistribution. We did not observe interactions between the storage method and the posterior washing of the samples. PRACTITIONER POINTS: We studied the influence of storage procedure on element content in aquatic mosses. The procedures were as follows: dried at room temperature, stored in refrigerator, and frozen. The procedures did not change the element contents, except for Hg in refrigerator. All the procedures seemed suitable, but refrigeration was the simplest one.

Diversity of Moss Species (Bryophyta) In Senggani Ravine Tourism Area, Tulungagung Regency

Moss plants (Bryophyta) are found in every habitat, and their presence in an ecosystem is controlled by environmental circumstances. The Senggani Ravine tourism area is a popular tourist attraction comprising a pine forest with extensive moss communities. This work aimed to assess the diversity of mosses (Bryophyta) in the Senggani Ravine tourism area for the first time. In June 2022, exploratory descriptive research of moss diversity was undertaken by a free walk around the Senggani Ravine tourism area from a predetermined position point (purposive sampling) using observation, documentation, literature study, and measurement of abiotic elements for data collection. Based on the results, twenty species of mosses were identified and can be divided into four classes, namely the Bryopsida, Polytrichopsida, Jungermanniopsida, and Marchantiopsida. The species identified were Barbulla indica, Fissidens purpusillus, Fissidens biformis, Fissidens biformis, Octoblepharum albidum, Rhizonium punctatum, Mnium hornum, Philonotis marchica, Fontinalis antipyretica, Hypnum cupressiform, Polytrichastrum formosum, Lejeunea flava, Lejeunea cavifolia, Bazzania prareupta, Bazzania vittata, Riccia junghuhniana, Marchantia emarginata, Marchantia polymorpha, Dumortiera hirsuta, and Lunularia cruciate. Abiotic factor measurements revealed that zone 3, which has a soil pH of 6, an air temperature of 24.1 C°, an 84% humidity level, and 200 Cd of light cm-1, is the most favorable area for moss growth. We can infer that the Senggani Ravine tourism area is still primarily undisturbed because the moss flora is still quite diverse and varied.

Macrophyte-Based Assessment of Upland Rivers: Bioindicators and Biomonitors

For the first time, a macrophyte-based assessment of ecological status was related to the accumulated heavy metals and trace elements (Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Zn) in aquatic plants. Three moss and two vascular plant species were applied as biomonitors: Fontinalis antipyretica Hedw., Leptodictyum riparium (Hedw.) Warnst., Platyhypnidium riparioides (Hedw.) Dixon, invasive Elodea canadensis Michx., and Myriophyllum spicatum L. Three streams were assessed as good at a high ecological status which correlated with low contamination based on calculated contamination factors (CFs) and metal pollution index (MPI). Two sites evaluated in moderate ecological status were revealed to be in heavy trace element contamination. The most significant was the accumulation of moss samples from the Chepelarska River under mining impact. Mercury exceeded the environmental quality standard (EQS) for biota in three of the studied upland river sites.

Ecological Preferences and Indication Potential of Freshwater Bryophytes-Insights from Croatian Watercourses.

A comprehensive survey of Croatian watercourses covering the whole of the national territory and investigating inherent watercourse heterogeneity was conducted to explore the ecological responses of the most frequent freshwater bryophytes with respect to water chemistry variables and land use within the catchment area. Direct multivariate ordination (CCA) of vegetation data paired with 18 environmental variables revealed that freshwater bryophytes and their assemblages were segregated along the gradients of water chemistry and the proportion of natural and urban area within the catchment. Generalized additive models (GAM) were employed to explore the ecological responses of individual species. The results showed that most of the investigated species preferred natural, clean, well-oxygenated watercourses, with low nutrient and organic matter content, as well as with low electrical conductivity. Species such as Palustriella falcata, Eucladium vertcillatum, Dichodontium flavescens and Jungermannia atrovirens had narrow ecological niches and were restricted to pristine watercourses, while the most frequent and widely distributed species, such as Fontinalis antipyretica, Rhynchostegium riparioides, Cratoneuron filicinum, Fissidens crassipes, Cinclidotus fontinaloides and C. riparius, had a wide ecological tolerance. Riccia fluitans and Leptodyctium riparium had wide ecological ranges, but with optima in hypereutrophic waters with high nutrient and organic content, as well as high electrical conductivity. Furthermore, these two species were frequently associated with a high share of intensive agriculture and a low share of natural land within the catchment.

Cyanotoxins associated with macrophytes in Berlin (Germany) water bodies – Occurrence and risk assessment

Fatal dog poisoning after uptake of neurotoxic cyanobacteria associated with aquatic macrophytes in Tegeler See (Berlin, Germany) raised concerns about critical exposure of humans, especially children, to cyanotoxins produced by macrophyte associated cyanobacteria during recreational activity. From 2017 to 2021 a total of 398 samples of macrophytes washed ashore at bathing sites located at 19 Berlin lakes were analysed for anatoxins, microcystins, and cylindrospermopsins, as were 463 water samples taken in direct proximity to macrophyte accumulations. Cyanotoxins were detected in 66 % of macrophyte samples and 50 % of water samples, with anatoxins being the most frequently detected toxin group in macrophyte samples (58 %) and cylindrospermopsins in water samples (41 %). Microcoleus sp. associated with the water moss Fontinalis antipyretica was identified as anatoxin producing cyanobacterium in isolated strains as well as in field samples from Tegeler See. Anatoxin contents in macrophyte samples rarely exceeded 1 μg/g macrophyte fresh weight and peaked at 9. 2 μg/g f.w. Based on established toxicological points of departure, a critical anatoxin content of macrophyte samples of 3 μg/g f.w. is proposed. Five samples, all taken in Tegeler See and all associated with the water moss Fontinalis antipyretica, exceeded this value. Contents and concentrations of microcystins and cylindrospermopsins did not reach critical levels. The potential exposure risks to anatoxins for children and dogs are assessed and recommendations are given.

Characterization and evolutionary diversification of the phospholipase D gene family in mosses.

Plant phospholipase D (PLD) exerts important roles in various biological processes, such as intracellular signaling and morphological development. Our knowledge about early land plant PLDs is still underdeveloped. In this study, we identified 84 PLD genes in six mosses, i.e., Physcomitrella patens, Ceratodon purpureus, Fontinalis antipyretica, Pleurozium schreberi, Sphagnum magellanicum, and Sphagnum fallax. These PLDs were classified into four clades (I–IV). We showed that PLD underwent rapid expansion in mosses. A total of six conserved domains and two core HKD motifs were detected. Structure analysis uncovered that the moss PLDs from within a clade generally exhibited similar exon-intron organization. Cis-elements prediction and expression analyses indicated that P. patens PLDs had key roles in stress responsiveness and plant development. Particularly, about half of the P. patens PLDs (e.g., PpPLD1, PpPLD2, and PpPLD5) were differentially expressed under biotic and abiotic stresses. We also determined the expression pattern of P. patens PLD genes in various tissues and at different stages of development. Although the moss, clubmoss, liverwort, and fern PLDs evolved largely under functional constraints, we found episodic positive selection in the moss PLDs, e.g., C. purpureus PLD2 and P. patens PLD11. We infer that the evolutionary force acting on the PLDs may have facilitated moss colonization of land. Our work provides valuable insights into the diversification of moss PLD genes, and can be used for future studies of their functions.

Suitability of aquatic mosses for biomonitoring micro/meso plastics in freshwater ecosystems

BackgroundMesoplastics (5–25 mm) and microplastics (0.001–5 mm) are emerging pollutants of great concern. However, reliable methods of monitoring these types of plastic in river ecosystems have not yet been established. The goal of this work was to evaluate, for the first time, the suitability of Fontinalis antipyretica as a biomonitor of meso- and micro-plastics in rivers. With this aim, native samples of the moss and devitalized moss clones, held inside the bags, were compared for the uptake of fluorescent polystyrene particles under laboratory conditions, and for retention of plastic debris in the field, in sites close to wastewater treatment plants.ResultsIn the laboratory experiment, the moss retained smaller microplastics, and a higher number of polystyrene meso and microplastics was counted in the moss bags than in the native moss.In the field study, the moss retained plastic debris chiefly in the form of fibres regardless of the capacity and flow rate of the wastewater treatment plants affecting each sampling site. The uniform morphology of moss clone seems to affect the retention of this type of pollutant. The FTIR analysis confirmed the particles entrapped by the moss bags as plastic, specifically polyethylene and polyamide type 6, among the most common plastic polymers detected in rivers.ConclusionsThe study findings highlighted the value of using uniform material, as the clone exhibited a greater accumulation efficiency with respect to the native moss. The mesh bags could act as selective filters and/or prevent the loss of adhering plastics. In the field, the bags favour plastic fibres retention despite the river flow. Finally, although FTIR is useful for the identification of plastic type, it is not very sensitive when small quantities of ground samples are used.

Environmental ranges discriminating between macrophytes groups in European rivers.

Riverine macrophytes form distinct species groups. Their occurrence is determined by environmental gradients, e.g. in terms of physico-chemistry and hydromorphology. However, the ranges of environmental variables discriminating between species groups (“discriminatory ranges”) have rarely been quantified and mainly been based on expert judgement, thus limiting options for predicting and assessing ecosystem characteristics. We used a pan-European dataset of riverine macrophyte surveys obtained from 22 countries including data on total phosphorus, nitrate, alkalinity, flow velocity, depth, width and substrate type. Four macrophyte species groups were identified by cluster analysis based on species’ co-occurrences. These comprised Group 1) mosses, such as Amblystegium fluviatile and Fontinalis antipyretica, Group 2) shorter and pioneer species such as Callitriche spp., Group 3) emergent and floating species such as Sagittaria sagittifolia and Lemna spp., and Group 4) eutraphent species such as Myriophyllum spicatum and Stuckenia pectinata. With Random Forest models, the ranges of environmental variables discriminating between these groups were estimated as follows: 100–150 μg L-1 total phosphorus, 0.5–20 mg L-1 nitrate, 1–2 meq L-1 alkalinity, 0.05–0.70 m s-1 flow velocity, 0.3–1.0 m depth and 20–80 m width. Mosses were strongly related to coarse substrate, while vascular plants were related to finer sediment. The four macrophyte groups and the discriminatory ranges of environmental variables fit well with those described in literature, but have now for the first time been quantitatively approximated with a large dataset, suggesting generalizable patterns applicable at regional and local scales.

Croatian freshwater bryoflora-diversity and distribution.

An extensive macrophyte field survey of running and standing waters was conducted from 2016 to 2021 at 786 sampling sites across Croatia as a part of the implementation of the Water Framework Directive. This survey is the first to present a comprehensive floristic catalogue of the freshwater bryoflora, along with an analysis of the distribution and diversity patterns on a national level. In all, 83 bryophyte species (68 mosses and 15 liverworts) were recorded in the 228 sites, with average species richness of 4.17 species per site. The most frequent species were Fontinalisantipyretica, Rhynchostegiumriparioides, Leptodictyumriparium and Cratoneuronfilicinum. The majority of the species encountered were rarely found, with over 70% of species recorded on less than 10 sampling sites and the majority of the species not being truly aquatic, rather being classified as facultative aquatics. The Dinaric Ecoregion, characterised by clean, cold, fast-flowing karstic rivers, especially in the Continental Subecoregion, supported higher freshwater bryophyte diversity than the lowland Pannonian Ecoregion, with mostly slow, eutrophic lowland watercourses with unstable sandy and gravelly alluvial sediments. Chorological comparison of Croatian eco- and subecoregions revealed the expected dominance of circumpolar and European elements, i.e. temperate chorotypes, as well as some biogeographical differences. The most frequent life forms were aquatic trailings and turfs. Amongst the recorded species, perennials and colonists were the most represented life strategies. The analysis of both the life-form and life-strategy spectra showed some differences amongst the Croatian regions, supporting the fact that the Dinaric Ecoregion provides more truly aquatic habitats and microhabitats suitable for the freshwater bryophytes, while in the Pannonian Ecoregion freshwater bryophytes dominantly inhabit the periodically submerged riparian zones, for example shaded lowland forest streams and rivulets or gently sloping margins of rivers and lakes.

Risk Assessment of Gypsum Amendment on Agricultural Fields: Effects of Sulfate on Riverine Biota.

Gypsum (CaSO4∙2H2O) amendment is a promising way of decreasing the phosphorus loading of arable lands, and thus preventing aquatic eutrophication. However, in freshwaters with low sulfate concentrations, gypsum‐released sulfate may pose a threat to the biota. To assess such risks, we performed a series of sulfate toxicity tests in the laboratory and conducted field surveys. These field surveys were associated with a large‐scale pilot exercise involving spreading gypsum on agricultural fields covering 18% of the Savijoki River (Finland) catchment area. The gypsum amendment in such fields resulted in approximately a four‐fold increase in the mean sulfate concentration for a 2‐month period, and a transient, early peak reaching approximately 220 mg/L. The sulfate concentration gradually decreased almost to the pregypsum level after 3 years. Laboratory experiments with Unio crassus mussels and gypsum‐spiked river water showed significant effects on foot movement activity, which was more intense with the highest sulfate concentration (1100 mg/L) than with the control. Survival of the glochidia after 24 and 48 h of exposure was not significantly affected by sulfate concentrations up to 1000 mg/L, nor was the length growth of the moss Fontinalis antipyretica affected. The field studies on benthic algal biomass accrual, mussel and fish density, and Salmo trutta embryo survival did not show gypsum amendment effects. Gypsum treatment did not raise the sulfate concentrations even to a level just close to critical for the biota studied. However, because the effects of sulfate are dependent on both the spatial and the temporal contexts, we advocate water quality and biota monitoring with proper temporal and spatial control in rivers within gypsum treatment areas. Environ Toxicol Chem 2022;41:108–121. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Interest of a multispecies approach in active biomonitoring: Application in the Meuse watershed

A biomonitoring approach based on a single model species cannot be representative of the contaminations impacts on the ecosystem overall. As part of the Interreg DIADeM program (“Development of an integrated approach for the diagnosis of the water quality of the River Meuse”), a study was conducted to establish the proof of concept that the use of a multispecies active biomonitoring approach improves diagnostic of aquatic systems. The complementarity of the biomarker responses was tested in four model species belonging to various ecological compartments: the bryophyte Fontinalis antipyretica, the bivalve Dreissena polymorpha, the amphipod Gammarus fossarum and the fish Gasterosteus aculeatus. The species have been caged upstream and downstream from five wastewater treatment plants (WWTPs) in the Meuse watershed. After the exposure, a battery of biomarkers was measured and results were compiled in an Integrated Biomarker Response (IBR) for each species. A multispecies IBR value was then proposed to assess the quality of the receiving environment upstream the WWTPs.The effluent toxicity was variable according to the caged species and the WWTP. However, the calculated IBR were high for all species and upstream sites, suggesting that the water quality was already downgraded upstream the WWTP. This contamination of the receiving environment was confirmed by the multispecies IBR which has allowed to rank the rivers from the less to the most contaminated. This study has demonstrated the interest of the IBR in the assessment of biological impacts of a point-source contamination (WWTP effluent) but also of the receiving environment, thanks to the use of independent references. Moreover, this study has highlighted the complementarity between the different species and has emphasized the interest of this multispecies approach to consider the variability of the species exposition pathway and sensibility as well as the mechanism of contaminants toxicity in the final diagnosis.

An efficient method to wash out the particulate matter trapped by aquatic mosses

Aquatic macrophytes capture trace elements from water and store them in their tissues, which make them good biomonitors for these substances. But they also capture suspended particulate matter that could bias the concentration measurements in the moss tissues. To avoid this problem, many biomonitoring studies using mosses include a washing step in the sample preparation procedure.However, no data is available on the quantities and variability of the particulate matter trapped by the moss, and the washing procedure has not been standardized between studies (washing time varied between 10 s to 30 min, divided in up to 7 washing steps). To study this, we extracted the particulate matter from samples of the moss Fontinalis antipyretica Hedw. collected in three rivers in NW Spain, washed them for 1, 2 and 5 min and collected the particulate matter extracted in each step. We also measured the contents of Al, As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn in the tissues of the washed and unwashed mosses. We made a second experiment washing the samples only for 15 + 45, 30 + 30 and 45 + 15 s to determine if shorter washing times were adequate for cleaning the moss. In a third experiment we estimated the contents of particulate matter in moss samples collected monthly over a two-year period in two rivers in the same area.The results showed that a) particulate matter can be as high as 40\\% of the total weight of the sample, and that there were large variations within and between sampling sites, and over time. These results showed the need of washing the samples prior to their analysis; b) that two washings are needed for the removal of most of the particulate matter, but that 30 + 30 s are long enough to clean the moss, and c) that a 20:1 (or larger) water:moss relation is recommended to avoid saturation of the washing water, which would reduce the efficiency of the procedure.We did not find significant losses of trace elements along the washing process, except for Cr in the three sampling sites, and Al, Fe and Pb in some of them. 1 min total washing time seemed the correct option both from the point of view of reducing the handling time of the samples and to prevent possible element losses.

Toxic effects of a mixture of five pharmaceutical drugs assessed using Fontinalis antipyretica Hedw.

The potential health risks associated with the pharmaceuticals released into the environment through effluents from sewage treatment plants have become a major cause for concern. Owing to the lack of effective indicators, monitoring the concentration of these pollutants in the aquatic environment is challenging. The aim of this study was to assess the toxicity of a mixture of five pharmaceutical drugs (paracetamol, carbamazepine, diclofenac, irbesartan, and naproxen) using the aquatic moss Fontinalis antipyretica as a bioindicator and bioaccumulator. We examined the effects of the drug mixture on the cellular antioxidant system, chlorophyll content, and morphological traits of F. antipyretica. The plant was exposed for 5 months to three concentrations of the mixture, including the environmental concentration (MX1), and 10- (MX10) and 100-times (MX100) the environmental concentration. The results showed that only carbamazepine and irbesartan were accumulated by the species. The bioconcentration level increased with exposure time, with the maximum uptake at the 4th month of exposure. The increase in bioaccumulation with exposure time was more evident in plants exposed to MX100. Analysis of the activity of antioxidant enzymes showed that superoxide dismutase (SOD, EC 1.15.1.1.) and catalase (EC 1.11.1.6.) were highly sensitive to the drug mixture. The activity of the enzymes was significantly higher in plants exposed to MX100; however, the activity of guaiacol peroxidase (GPX, EC 1.11.1.7.) was not significantly affected. Plants exposed to MX10 and MX100 had significantly lower total chlorophyll content and chlorophyll a/b ratio compared with those of plants in the control group; however, photosynthetic activity was restored after 5 months of exposure. The morphological characteristics of F. antipyretica were less sensitive to the treatment conditions.