Mining is responsible of releasing trace elements to the environment with potential negative effects on wildlife. Traditionally, wildlife exposure assessment has been developed by analyzing mainly environmental compartments or internal tissues. Nowadays, the use of non-destructive matrices such as hair or feathers has increased. Nevertheless, its use in free-living terrestrial mammals or in birds other than raptors or passerines is less frequent. The main objective of our study was to determine the potential for hair and feathers in a rabbit and bird species to be used as non-invasive proxy tissues for assessing internal metal concentrations at polluted sites from mining. We tested whether hair of European rabbit (Oryctolagus cuniculus) and feathers of red-legged partridge (Alectoris rufa) can be used as non-destructive biological monitoring tools of both essential (Cu, Zn) and non-essential (Pb, Cd, As) trace elements in a currently active copper mining site. We found significant different concentrations, particularly in non-essential elements, between reference area and mining site. Non-essential elements Pb and Cd showed higher correlations between tissues and hair/feathers, while few significant patterns were observed for essential elements such as Cu and Zn. Although feathers showed lower levels of correlation with internal tissues than hair, both could be useful as non-destructive biological monitoring tools. Further tissues, and more importantly, hair and feathers allowed discrimination between polluted and reference sites to indicate bioavailability and pollution status. In addition, hair and feathers can be used in monitoring pollution of an active mining site, being specially interesting for biomonitoring a certain period of time in the event of a particular episode of pollution, in addition to the chronic exposure. As occurred with hair in rabbits, feathers seem to be a good compartment to detect differences between a potential polluted area, such the surrounding of an active mine site, and a non-polluted area.

Mining is responsible of releasing trace elements to the environment with potential negative effects on wildlife. Traditionally, wildlife exposure assessment has been developed by analyzing mainly environmental compartments or internal tissues. Nowadays, the use of non-destructive matrices such as hair or feathers has increased. Nevertheless, its use in free-living terrestrial mammals or in birds other than raptors or passerines is less frequent. The main objective of our study was to determine the potential for hair and feathers in a rabbit and bird species to be used as non-invasive proxy tissues for assessing internal metal concentrations at polluted sites from mining. We tested whether hair of European rabbit (Oryctolagus cuniculus) and feathers of red-legged partridge (Alectoris rufa) can be used as non-destructive biological monitoring tools of both essential (Cu, Zn) and non-essential (Pb, Cd, As) trace elements in a currently active copper mining site. We found significant different concentrations, particularly in non-essential elements, between reference area and mining site. Non-essential elements Pb and Cd showed higher correlations between tissues and hair/feathers, while few significant patterns were observed for essential elements such as Cu and Zn. Although feathers showed lower levels of correlation with internal tissues than hair, both could be useful as non-destructive biological monitoring tools. Further tissues, and more importantly, hair and feathers allowed discrimination between polluted and reference sites to indicate bioavailability and pollution status. In addition, hair and feathers can be used in monitoring pollution of an active mining site, being specially interesting for biomonitoring a certain period of time in the event of a particular episode of pollution, in addition to the chronic exposure. As occurred with hair in rabbits, feathers seem to be a good compartment to detect differences between a potential polluted area, such the surrounding of an active mine site, and a non-polluted area. | This work was supported by funding from Fundación Migres through a collaboration project with Cobre Las Cruces, S.A. We would like to thank technicians from Fundacion Migres for their help during sampling. | Peer reviewed

Anthropogenic fibers, gathering synthetic fibers, artificial fibers and natural fibers are ubiquitous in the natural environment. Tremendous concentrations of anthropogenic fibers were previously measured in the tropical Saigon River (Vietnam), i.e. a river impacted by textile and apparel industries. In the present study, we want to examine the role of contrasted seasonal variation (e.g., dry and rainy seasons), via the rainfall and monthly water discharges, and of water's physico-chemical conditions on the concentrations of anthropogenic fibers in the surface water. The one year and half monthly survey evidenced that concentrations of anthropogenic fibers varied from 22 to 251 items L⁻¹ and their variations were not related to rainfall, water discharge or abiotic factors. However, their color and length distribution varied monthly suggesting variations in sources and sinks. Based on the 2017 survey, we estimated an annual emission of anthropogenic fibers from the river to the downstream coastal zone of 115–164 × 10¹² items yr⁻¹.

International audience | Anthropogenic fibers, gathering synthetic fibers, artificial fibers and natural fibers are ubiquitous in the natural environment. Tremendous concentrations of anthropogenic fibers were previously measured in the tropical Saigon River (Vietnam), i.e. a river impacted by textile and apparel industries. In the present study, we want to examine the role of contrasted seasonal variation (e.g., dry and rainy seasons), via the rainfall and monthly water discharges, and of water's physico-chemical conditions on the concentrations of anthropogenic fibers in the surface water. The one year and half monthly survey evidenced that concentrations of anthropogenic fibers varied from 22 to 251 items L-1 and their variations were not related to rainfall, water discharge or abiotic factors. However, their color and length distribution varied monthly suggesting variations in sources and sinks. Based on the 2017 survey, we estimated an annual emission of anthropogenic fibers from the river to the downstream coastal zone of 115e164 × 10 12 items yr-1.

The presence of microplastics in aquatic ecosystems has recently received increased attention. Small plastic particles may resemble natural food items of larval fish and other aquatic organisms, and create strong selective pressures on the feeding traits in exposed populations. Here, we examined if larval ingestion of 90 μm polystyrene microspheres, in the presence of zooplankton (Artemia nauplii, mean length = 433 μm), shows adaptive variation in the European whitefish (Coregonus lavaretus). A full-factorial experimental breeding design allowed us to estimate the relative contributions of male (sire) and female (dam) parents and full-sib family variance in early feeding traits, and also genetic (co)variation between these traits. We also monitored the magnitude of intake and elimination of microplastics from the alimentary tracts of the larvae. In general, larval whitefish ingested small numbers of microplastics (mean = 1.8, range = 0–26 particles per larva), but ingestion was marginally affected by the dam, and more strongly by the full-sib family variation. Microsphere ingestion showed no statistically significant additive genetic variation, and thus, no heritability. Moreover, microsphere ingestion rate covaried positively with the ingestion of Artemia, further suggesting that larvae cannot adaptively avoid microsphere ingestion. Together with the detected strong genetic correlation between food intake and microplastic intake, the results suggest that larval fish do not readily possess additive genetic variation that would help them to adapt to the increasing pollution by microplastics. The conflict between feeding on natural food and avoiding microplastics deserves further attention.

2020

Wastewater discharges from dairy industries can cause a range of harmful effects in aquatic ecosystems, including a decline in biodiversity due to species evasion. Therefore, it is important to know the purification potential of rivers for the removal of pollutants released in dairy wastewater (DWW). The hypothesis adopted in this work was that the release of DWW into stretches of the Ribeirão dos Pombos River (São Paulo State, Brazil) might trigger an avoidance response, resulting in fish migrating to other regions, with the response being greater when the self-cleaning potential of the river is smaller. Therefore, the goals of the present study were to: (i) investigate how land use and seasonality of the rainfall regime influence the quality of the water in different areas of the river (P1: river source; P2: urban region; P3: rural region); (ii) assess the potential of the river to purify DWW; and (iii) evaluate the potential toxicity and repellency of DWW to the freshwater fish Danio rerio, using acute toxicity (mortality) and non-forced avoidance tests, respectively. P1 was shown to be the most preserved area. The chemical composition of the river varied seasonally, with higher concentrations of Cl⁻ and SO₄²⁻ at P3 during the rainy period. The river purification potential for DWW was higher at P2, due to greater microbiological activity (associated with higher BOD). The DWW was more acutely toxic in water from P2. The avoidance response was strongly determined by the concentration of DWW, especially for water from P2. The high capacity for self-cleaning at P2 did not seem sufficient to maintain the stability of the ecosystem. Finally, the non-forced exposure system proved to be a suitable approach that can assist in predicting how contaminants may affect the spatial distributions of organisms.

Wastewater discharges from dairy industries can cause a range of harmful effects in aquatic ecosystems, including a decline in biodiversity due to species evasion. Therefore, it is important to know the purification potential of rivers for the removal of pollutants released in dairy wastewater (DWW). The hypothesis adopted in this work was that the release of DWW into stretches of the Ribeirão dos Pombos River (São Paulo State, Brazil) might trigger an avoidance response, resulting in fish migrating to other regions, with the response being greater when the self-cleaning potential of the river is smaller. Therefore, the goals of the present study were to: (i) investigate how land use and seasonality of the rainfall regime influence the quality of the water in different areas of the river (P1: river source; P2: urban region; P3: rural region); (ii) assess the potential of the river to purify DWW; and (iii) evaluate the potential toxicity and repellency of DWW to the freshwater fish Danio rerio, using acute toxicity (mortality) and non-forced avoidance tests, respectively. P1 was shown to be the most preserved area. The chemical composition of the river varied seasonally, with higher concentrations of Cl- and SO42- at P3 during the rainy period. The river purification potential for DWW was higher at P2, due to greater microbiological activity (associated with higher BOD). The DWW was more acutely toxic in water from P2. The avoidance response was strongly determined by the concentration of DWW, especially for water from P2. The high capacity for self-cleaning at P2 did not seem sufficient to maintain the stability of the ecosystem. Finally, the non-forced exposure system proved to be a suitable approach that can assist in predicting how contaminants may affect the spatial distributions of organisms. | Financial support for this work was provided by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant #14/22581–8). This study was financed in part by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES, Finance Code 001). CVMA is grateful to the Spanish Ministry of Science and Innovation for a Ramón y Cajal contract (RYC-2017-22324). This study was performed within the framework of the cooperation project MultiCecotox (COOPB20444; i-COOP2019 program from CSIC). | Peer reviewed

Marine fishes are exposed to great human-induced alterations due to the indiscriminate discharges into the sea, increasing marine pollution. For this study, 324 specimens of Sardina pilchardus from the Canary Islands were analized during a period of 2 years (June 2016 to May 2018). The concentration of 11 metals and trace elements (Al, B, Cd, Cr, Cu, Fe, Li, Ni, Pb, V and Zn) was determined in each individual using the Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) technique. Statistical analyses were carried out considering the following factors: oceanographic season, maturity of the gonads in the seasons, size of the specimens. Sardina pilchardus specimens captured in the hot season presented higher Ni, Li and Pb levels than the specimens caught in the cold season. Immature specimens had higher concentration in more metals than the mature specimens. This fact may be due to the fact that these specimens require a much higher metabolic rate due to their growth and do not detoxify like mature specimens. Significant differences were found in the concentration of metals in all the analyzes performed.

Marine fishes are exposed to great human-induced alterations due to the indiscriminate discharges into the sea, increasing marine pollution. For this study, 324 specimens of Sardina pilchardus from the Canary Islands were analized during a period of 2 years (June 2016 to May 2018). The concentration of 11 metals and trace elements (Al, B, Cd, Cr, Cu, Fe, Li, Ni, Pb, V and Zn) was determined in each individual using the Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) technique. Statistical analyses were carried out considering the following factors: oceanographic season, maturity of the gonads in the seasons, size of the specimens. Sardina pilchardus specimens captured in the hot season presented higher Ni, Li and Pb levels than the specimens caught in the cold season. Immature specimens had higher concentration in more metals than the mature specimens. This fact may be due to the fact that these specimens require a much higher metabolic rate due to their growth and do not detoxify like mature specimens. Significant differences were found in the concentration of metals in all the analyzes performed.

Marine fishes are exposed to great human-induced alterations due to the indiscriminate discharges into the sea, increasing marine pollution. For this study, 324 specimens of Sardina pilchardus from the Canary Islands were analized during a period of 2 years (June 2016 to May 2018). The concentration of 11 metals and trace elements (Al, B, Cd, Cr, Cu, Fe, Li, Ni, Pb, V and Zn) was determined in each individual using the Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) technique. Statistical analyses were carried out considering the following factors: oceanographic season, maturity of the gonads in the seasons, size of the specimens. Sardina pilchardus specimens captured in the hot season presented higher Ni, Li and Pb levels than the specimens caught in the cold season. Immature specimens had higher concentration in more metals than the mature specimens. This fact may be due to the fact that these specimens require a much higher metabolic rate due to their growth and do not detoxify like mature specimens. Significant differences were found in the concentration of metals in all the analyzes performed. | Sí

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and several other environment/food-borne toxic compounds induce their toxicity via the aryl hydrocarbon receptor (AhR). AhR is also modulated by various endogenous ligands e.g. highly potent tryptophan (Trp)-derivative FICZ (6-formylindolo[3,2-b]carbazole) and natural ligands abundant in the human diet e.g. polyphenols. Therefore, evaluating AhR species-specific responses is crucial for understanding AhR physiological functions, establishing risk assessments, and exploring the applicability of AhR mediators in drug and food industry towards human-based usages. We studied AhR transactivation of FICZ/TCDD in vitro in a time-dependent and species-specific manner using dioxin responsive luciferase reporter gene assays derived from rat (DR-H4IIE) and human (DR-HepG2) hepatoma cells. We observed for the first time that FICZ potency was similar in both cell lines and was 40 times higher than TCDD in DR-HepG2 cells. Depleting Trp-derivative endogenously produced ligands by using culture medium without Trp, resulted in 3-fold higher AhR activation upon adding FICZ in DR-H4IIE cells, in contrast to DR-HepG2 cells which revealed a fast degradation of FICZ induction from 10 h post-exposure to complete disappearance after 24 h. Seven polyphenols and a mixture thereof, chosen based on commercially recommended doses and adjusted to human realistic exposure, caused rat and human species-specific AhR responses. Two isoflavones (daidzein and genistein) induced rat AhR synergistic effects with FICZ and/or TCDD, while quercetin, chrysin, curcumin, resveratrol, and the mixture exerted a strong inhibitory effect on the human AhR. Strikingly, resveratrol and quercetin at their realistic nanomolar concentrations acted additively in the mixture to abolish human AhR activation induced by various TCDD concentrations. Taken together, these results illustrate the species-specific complexity of AhR transcriptional activities modulated by various ligands and highlight the need for studies of human-based approaches.

peer reviewed | Protected

Considering the increasing use of Lithium (Li) and the necessity to fulfil this demand, labile Li occurrence in the environment will be enhanced. Thus, additional research is needed regarding the presence of this element in marine environment and its potential toxic impacts towards inhabiting wildlife. The aim of the present study was to evaluate Li toxicity based on the exposure of Mytilus galloprovincialis to this metal, assessing the biochemical changes related with mussels’ metabolism, oxidative stress and neurotoxicity. For this, organisms were exposed to different Li concentrations (100, 250, 750 μg/L) for 28 days. The results obtained clearly demonstrated that Li lead to mussels’ metabolism depression. The present study also revealed that, especially at the highest concentrations, antioxidant and biotransformation enzymes were not activated, leading to the occurrence of lipid peroxidation and loss of redox homeostasis, with increased content in oxidized glutathione in comparison to the reduced form. Furthermore, after 28 days, higher Li exposure concentrations induced neurotoxic effects in mussels, with a decrease in acetylcholinesterase enzyme activity. The responses observed were closely related with Li concentrations in mussels’ tissues, which were more pronounced at higher exposure concentrations. Such results highlight the potential toxic effects of Li to marine species, which may even be higher under predicted climate changes and/or in the presence of other pollutants.

Considering the increasing use of Lithium (Li) and the necessity to fulfil this demand, labile Li occurrence in the environment will be enhanced. Thus, additional research is needed regarding the presence of this element in marine environment and its potential toxic impacts towards inhabiting wildlife. The aim of the present study was to evaluate Li toxicity based on the exposure of Mytilus galloprovincialis to this metal, assessing the biochemical changes related with mussels' metabolism, oxidative stress and neurotoxicity. For this, organisms were exposed to different Li concentrations (100, 250, 750 μg/L) for 28 days. The results obtained clearly demonstrated that Li lead to mussels' metabolism depression. The present study also revealed that, especially at the highest concentrations, antioxidant and biotransformation enzymes were not activated, leading to the occurrence of lipid peroxidation and loss of redox homeostasis, with increased content in oxidized glutathione in comparison to the reduced form. Furthermore, after 28 days, higher Li exposure concentrations induced neurotoxic effects in mussels, with a decrease in acetylcholinesterase enzyme activity. The responses observed were closely related with Li concentrations in mussels' tissues, which were more pronounced at higher exposure concentrations. Such results highlight the potential toxic effects of Li to marine species, which may even be higher under predicted climate changes and/or in the presence of other pollutants. | published

Antibiotics, such as sulfonamides (SAs), have recently raised concern as wastewater treatment plants (WWTPs) partly remove them, and thus, SAs continuously enter the aquifers. In this context, the aims of this work are to (1) investigate the temporal evolution of SAs and metabolites in an urban aquifer recharged by a polluted river; (2) identify the potential geochemical processes that might affect SAs in the river-groundwater interface and (3) evaluate the ecological and human health risk assessment of SAs. To this end, 14 SAs and 4 metabolites were analyzed in river and urban groundwater from the metropolitan area of Barcelona (NE, Spain) in three different sampling campaigns. These substances had a distinct behavior when river water, which is the main recharge source, infiltrates the aquifer. Mixing of the river water recharge into the aquifer drives several redox reactions such as aerobic respiration and denitrification. This reducing character of the aquifer seemed to favor the natural attenuation of some SAs as sulfamethoxazole, sulfapyridine, and sulfamethizole. However, most of the SAs detected were not likely to undergo degradation and adsorption because their concentrations were constant along groundwater flow path. In fact, the intensity of SAs adsorption is low as the retardation factors are close to 1 at average groundwater pH of 7.2 for most SAs.Finally, risk quotients (RQs) are used to evaluate the ecological and human health risks posed by single and mixture of SAs in river water and groundwater, respectively. Life-stage RQs of the SAs detected in groundwater for the 8 age intervals were low, indicating that SAs and their mixture do not pose any risk to human beings. Concerning the environmental risk assessment, SAs do not pose any risk for algae, fish and crustaceans as the RQs evaluated are further lower than 0.1.

Antibiotics, such as sulfonamides (SAs), have recently raised concern as wastewater treatment plants (WWTPs) partly remove them, and thus, SAs continuously enter the aquifers. In this context, the aims of this work are to (1) investigate the temporal evolution of SAs and metabolites in an urban aquifer recharged by a polluted river; (2) identify the potential geochemical processes that might affect SAs in the river-groundwater interface and (3) evaluate the ecological and human health risk assessment of SAs. To this end, 14 SAs and 4 metabolites were analyzed in river and urban groundwater from the metropolitan area of Barcelona (NE, Spain) in three different sampling campaigns. These substances had a distinct behavior when river water, which is the main recharge source, infiltrates the aquifer. Mixing of the river water recharge into the aquifer drives several redox reactions such as aerobic respiration and denitrification. This reducing character of the aquifer seemed to favor the natural attenuation of some SAs as sulfamethoxazole, sulfapyridine, and sulfamethizole. However, most of the SAs detected were not likely to undergo degradation and adsorption because their concentrations were constant along groundwater flow path. In fact, the intensity of SAs adsorption is low as the retardation factors are close to 1 at average groundwater pH of 7.2 for most SAs. Finally, risk quotients (RQs) are used to evaluate the ecological and human health risks posed by single and mixture of SAs in river water and groundwater, respectively. Life-stage RQs of the SAs detected in groundwater for the 8 age intervals were low, indicating that SAs and their mixture do not pose any risk to human beings. Concerning the environmental risk assessment, SAs do not pose any risk for algae, fish and crustaceans as the RQs evaluated are further lower than 0.1. | A.J acknowledges the support from the Secretary for Universities and Research of the Ministry of Economy and Knowledge of the Government of Catalonia and the Marie Sklodowska-Curie COFUND of the programme H2020 (BP3, contract number 801370). M.S.D.C acknowledges the Generalitat de Catalunya (Water and Soil Quality Unit, 2017-SGR-1404) and the Spanish Ministry of Science and Innovation (Project CEX 2018-000794-S). E.V.S would like to thank the European Commission, the Spanish Foundation for Science & Technology (FECYT) and State Research Agency (AEI)) for funding in the frame of the collaborative international consortium (URBANWAT) financed under the 2018 Joint call of the WaterWorks2017 ERA-NET Cofund. This ERA-NET is an integral part of the activities developed by the Water JPI. Additionally E.V.S also thank the Ministry of Science, Innovation and Universities, for funding the project UNBIASED (Ref: RTI 2018-097346-B-I00) under the 2018 call of the “Proyectos de I + D Retos Investigación. | Peer reviewed

The Tharsis mine is presently abandoned, but the past intense exploitation has left large dumps and other sulphide-rich mining wastes in the area generating acid mine drainages (AMD). The main goal of this work is to study the effect of hydrogeochemical processes, hydrological regime and the waste typology on the physicochemical parameters and dissolved concentrations of pollutants in a deeply AMD-affected zone. Extreme leachates are produced in the area, reaching even negative pH and concentrations of up to 2.2 g/L of As and 194 g/L of Fe. The results of the comparison of ore grades of sulphide deposits with dissolved concentrations in waters shows that Pb is the least mobile element in dissolution probably due to the precipitation of Pb secondary minerals and/or its coprecipitation on Fe oxyhydroxysulphates. Arsenic, Cr, and V are also coprecipitated with Fe minerals. Seasonal patterns in metal contents were identified: elements coming from the host rocks, such as Al, Mn and Ni, show their maximum values in the dry period, when dilution with freshwater is lower and the interaction of water-rock processes and evaporation is higher. On the other hand, As, Cr, Fe, Pb and V show minimum concentrations in the dry period due to intense Fe oxyhydroxysulphate precipitation. In this sense, large sulphide rich waste heaps would be a temporal sink of these elements (i.e. Pb, As, Cr and V) in the dry period, and a significant source upon intense rainfalls.

The Tharsis mine is presently abandoned, but the past intense exploitation has left large dumps and other sulphide-rich mining wastes in the area generating acid mine drainages (AMD). The main goal of this work is to study the effect of hydrogeochemical processes, hydrological regime and the waste typology on the physicochemical parameters and dissolved concentrations of pollutants in a deeply AMD-affected zone. Extreme leachates are produced in the area, reaching even negative pH and concentrations of up to 2.2 g/L of As and 194 g/L of Fe. The results of the comparison of ore grades of sulphide deposits with dissolved concentrations in waters shows that Pb is the least mobile element in dissolution probably due to the precipitation of Pb secondary minerals and/or its coprecipitation on Fe oxyhydroxysulphates. Arsenic, Cr, and V are also coprecipitated with Fe minerals. Seasonal patterns in metal contents were identified: elements coming from the host rocks, such as Al, Mn and Ni, show their maximum values in the dry period, when dilution with freshwater is lower and the interaction of water-rock processes and evaporation is higher. On the other hand, As, Cr, Fe, Pb and V show minimum concentrations in the dry period due to intense Fe oxyhydroxysulphate precipitation. In this sense, large sulphide rich waste heaps would be a temporal sink of these elements (i.e. Pb, As, Cr and V) in the dry period, and a significant source upon intense rainfalls. | This work was supported by the Spanish Ministry of Economic and Competitiveness through the projects CGL2016-78783-C2-1-R (SCYRE) and by H2020 European Institute of Innovation and Technology (EIT RawMaterials) through the projects Modular recovery process services for hydrometallurgy and water treatment (MORECOVERY). The authors thank to Prof. Yong Sik Ok (Associate Editor) and five different reviewers for their helpful comments that notably improved the quality of the manuscript. | Peer reviewed

Freshwater planarians have been gaining relevance as experimental animals for numerous research areas given their interesting features, such as high regeneration potential, shared features with the vertebrates’ nervous system or the range of endpoints that can be easily evaluated in response to contaminants. Ecotoxicological research using these animals has been steadily increasing in the past decades, as planarians’ potentialities for this research area are being recognized. In this work, we used polycyclic aromatic hydrocarbons (PAHs) as model contaminants and evaluated effects of exposure to phenanthrene, pyrene and benzo[a]pyrene (B[a]P) in planarians. The freshwater planarian Girardia tigrina was chosen and mortality, cephalic regeneration (during and post-exposure), behavioral endpoints and presence of PAHs in tissues, were evaluated. Mortality was only observed in planarians exposed to phenanthrene, with an estimated LC₅₀ of 830 μg L⁻¹. Results indicate that planarian behavioral endpoints were very sensitive in response to sub-lethal concentrations of PAHs, showing a greater sensitivity towards B[a]P and pyrene. Briefly, post-exposure locomotion and post-exposure feeding were significantly impaired by sub-lethal concentrations of all compounds, whereas regeneration of photoreceptors was only significantly delayed in planarians exposed to pyrene. Moreover, levels of PAH-type compounds in planarian tissues followed a concentration-dependent increase, showing uptake of compounds from experimental solutions. The present results highlight the importance of studying alternative and complementary endpoints, such as behavior, not only because these may be able to detect effects at lower levels of contamination, but also due to their ecological relevance. The simplicity of evaluating a wide range of responses to contaminants further demonstrates the utility of freshwater planarians for ecotoxicological research.

Freshwater planarians have been gaining relevance as experimental animals for numerous research areas given their interesting features, such as high regeneration potential, shared features with the vertebrates' nervous system or the range of endpoints that can be easily evaluated in response to contaminants. Ecotoxicological research using these animals has been steadily increasing in the past decades, as planarians' potentialities for this research area are being recognized. In this work, we used polycyclic aromatic hydrocarbons (PAHs) as model contaminants and evaluated effects of exposure to phenanthrene, pyrene and benzo[a]pyrene (B[a]P) in planarians. The freshwater planarian Girardia tigrina was chosen and mortality, cephalic regeneration (during and post-exposure), behavioral endpoints and presence of PAHs in tissues, were evaluated. Mortality was only observed in planarians exposed to phenanthrene, with an estimated LC50 of 830 μg L-1. Results indicate that planarian behavioral endpoints were very sensitive in response to sub-lethal concentrations of PAHs, showing a greater sensitivity towards B[a]P and pyrene. Briefly, post-exposure locomotion and post-exposure feeding were significantly impaired by sub-lethal concentrations of all compounds, whereas regeneration of photoreceptors was only significantly delayed in planarians exposed to pyrene. Moreover, levels of PAH-type compounds in planarian tissues followed a concentration-dependent increase, showing uptake of compounds from experimental solutions. The present results highlight the importance of studying alternative and complementary endpoints, such as behavior, not only because these may be able to detect effects at lower levels of contamination, but also due to their ecological relevance. The simplicity of evaluating a wide range of responses to contaminants further demonstrates the utility of freshwater planarians for ecotoxicological research. | published