Cytogenotoxicity Screening of Urban and Rural Marshes: An Integrated In Vivo Approach Coupling Fish and Plant-Based Tests Adapted for Low-Income Countries.

Effects of anthropogenic activities such as urbanization, population growth, and agriculture on water quality are major concerns particularly in low-income countries where water quality monitoring can be challenging. The purpose of the present study was to evaluate the cytogenotoxic potential of water from urban and rural Malagasy marshes, coupling a fish (Nile tilapia, Oreochromis niloticus) and a plant (Allium cepa) species as bioindicators. The fish and plants were exposed for 72 h to water sampled in the two locations investigated. Using the comet assay on fish erythrocytes, DNA strand breaks were assessed, while mitotic index and nucleolar alterations were estimated in cells of the plant root apex. Comet assays revealed significant DNA strand breaks to fish erythrocytes in both the marshes investigated while the mitotic index and nucleolar characteristics in the roots of A. cepa mainly highlighted potential cytotoxicity in the urban marsh. Our results demonstrate the advantages of coupling in vivo biological test systems to screen potential cytogenotoxicity of surface water in low-income countries where comprehensive data sets of aquatic contaminants are often lacking. Environ Toxicol Chem 2023;42:1266-1275. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Stannous chloride as a tool for mercury stripping in contaminated streams: Experimental assessment of toxicity in an invertebrate model species.

The chronic toxicity of an innovative Hg water treatment system using tin (Sn) (II) chloride (SnCl2) followed by air stripping was assessed through measurements of survival, growth, and reproduction rate in the freshwater cladoceran Ceriodaphnia dubia, a model species for toxicity testing. We first calculated the concentrations of Hg causing 25% reduction in survival and reproduction (Lethal or Inhibition Concentrations, or LC25 and IC25, for survival and reproduction, respectively) through exposure to aqueous Hg at concentrations ranging from 0 to 25,000 ng L-1. Then, we treated media (DMW and natural stream water) contaminated with Hg at LC25 and IC25 concentrations with SnCl2 at a Sn:Hg stoichiometric ratio of 8:1 and air stripping and exposed C. dubia to this Sn-amended media. Our results showed that Hg significantly affected survival, reproduction rates and impaired growth. SnCl2-treatment removed 100% of the Hg from the media at all concentrations tested with no deleterious effects on survival, growth and reproduction. Our results confirmed the efficacy of SnCl2 in removing aqueous Hg from stream water and showed that the added Sn did not impact C. dubia at the concentrations tested, supporting the suitability of SnCl2-based treatments in appropriate Hg-contaminated environments.

Diet variably affects the trophic transfer of trace elements in the oyster Crassostrea gigas.

Although it has been shown that trophic transfer of trace elements in oysters can be influenced by the diet, most of the studies investigating the ability of oysters to bioaccumulate trace elements from their diet are based on experiments using phytoplankton alone. Wild oysters feed also on large bacteria, ciliates or detritic organic matter. The present study aimed at examining the influence of food quality on the assimilation efficiency (AE) of trace elements in the Pacific cupped oyster Crassostrea gigas. Oysters were exposed via their food to the radiotracers of essential (57Co, 54Mn and 65Zn) and non-essential (110mAg, 241Am and 109Cd) trace elements under different diets (protozoan ciliates Uronema marinum and diatoms Thalassiosira pseudonana). Significant differences were found only for Ag and 241Am, with lower AEs measured in oysters fed with ciliates than in individuals fed with diatoms (Ag: 54 ± 3% vs. 67 ± 4% and 241Am: 62 ± 4% vs. 76 ± 4%). Interestingly, no significant difference was found among estimated depuration rates (kel) for all trace elements ingested with the two diets tested. These findings indicate that the differences observed are driven by the digestion process, presumably due to difference of bioavailability of trace elements dependent on the quality of the food ingested.

Comparing Trace Element Bioaccumulation and Depuration in Snails and Mayfly Nymphs at a Coal Ash-Contaminated Site.

We examined the bioaccumulation of essential (Cu, Fe, Se, and Zn) and nonessential (As and Hg) trace elements in 2 aquatic invertebrate species (adult snails and mayfly nymphs) with different feeding habits at the site of a coal ash spill. Differences in food web pathway, exposure concentrations, and biological processing affected bioaccumulation patterns in these species. Mayflies had higher body burdens, but snails had higher retention of most elements studied. Environ Toxicol Chem 2020;39:2437-2449. © 2020 SETAC.

Trophic transfer of trace elements in a euryhaline fish, the turbot Scophthalmus maximus: Contrasting effects of salinity on two essential elements.

Trace elements can be accumulated from coastal environment by aquatic organisms from their food and be transferred throughout the food webs. Studying the effects of salinity on the trophic transfer of trace elements in a euryhaline fish, able to deal with large variations in salinity, is therefore key to understand their dynamics in aquatic environments. In this context, we investigated the potential influence of salinity on the trophic transfer of two essential elements (Mn and Zn) in the euryhaline fish, the turbot Scophthalmus maximus using radiotracer techniques. After acclimation to three salinities (10, 25 and 38), turbots were fed with radiolabelled pellets (54Mn and 65Zn). Kinetic parameters of depuration were determined after a 21-d period and trophic transfer factors were calculated. Trophic transfer of Mn at the highest salinity was significantly lower than for the other conditions whereas salinity did not significantly influenced Zn trophic transfer. Differences in the processes involved in the regulation (homeostasis) of the two tested trace elements may explain the contrasting influence of seawater salinity for Mn and Zn.

Influence of food (ciliate and phytoplankton) on the trophic transfer of inorganic and methyl-mercury in the Pacific cupped oyster Crassostrea gigas.

Diet is an important route of mercury (Hg) uptake in marine organisms. Trophic transfer of Hg throughout the food webs may be influenced by various factors, including diet and Hg speciation. Bivalves such as oysters are widely used as bioindicators of trace element pollution such as Hg. Nevertheless, our current knowledge regarding their ability to accumulate Hg from their diet is mainly based on experiments performed using phytoplankton. In their natural environment, oysters feed on a variety of food items including ciliates, detritus, in addition to phytoplankton. The present study aimed at examining the influence of diet composition on the trophic transfer of inorganic Hg (iHg) and methylmercury (MeHg) in the Pacific cupped oyster Crassostrea gigas. The pulse-chase feeding method was used with two radiolabeled food items: a heterotrophic protist (Uronema marinum) and a phytoplanktonic diatom (Thalassiosira pseudonana). Depuration of dietary Hg in the oysters was followed for 50 d. Kinetic parameters including assimilation efficiency (AE) and efflux rate constant (ke) were calculated. Our results showed that oysters fed on ciliates assimilated 96 ±â€¯1% and 31 ±â€¯2% of the ingested MeHg and iHg, respectively whereas these elements were similarly assimilated in the oysters fed on phytoplankton (78 ±â€¯3% and 86 ±â€¯4% for MeHg and iHg, respectively). Mercury assimilation in oyster is thus diet dependent (significant differences in AE, p < 0.05), metal species-dependent and likely resulting from variations in Hg bioavailability in the two food items tested and a gut passage time-dependent of the ingested matrix.

Radiocesium accumulation in aquatic organisms: A global synthesis from an experimentalist's perspective.

A better understanding of the fate of radiocesium in aquatic organisms is essential for making accurate assessments of potential impacts of radiocesium contamination on ecosystems and human health. Studies of the accumulation of 134Cs, 136Cs and 137Cs in diverse biota have been the subject of many field investigations; however, it may often be difficult to understand all the mechanisms underlying the observations reported. To complement field investigations, laboratory experiments allow better understanding the observations and predicting dynamics of Cs within aquatic ecosystems by accurately assessing bioaccumulation of Cs in living organisms. The present review summarizes selected relevant laboratory studies carried out on Cs bioaccumulation in aquatic organisms over a period of more than 60 years. To date, 125 experimental studies have been carried out on 227 species of aquatic organisms since 1957. The present review provides a synthesis of the existing literature by highlighting major findings and identifying gaps of key information that need to be further addressed in future works on this topic. Thus, influences of some environmental parameters such as water chemistry both for marine and freshwater ecosystems, and biotic factors such as the life-stages and size of the organisms on radiocesium bioaccumulation should be examined and become priority topics for future research on Cs accumulation in aquatic organisms.

Allometric relationship in the bioaccumulation of radionuclides (134Cs &241Am) and delineation of contamination pathways (food and seawater) in bloody cockle Anadara senilis using radiotracer techniques.

The uptake and depuration kinetics of 134Cs and 241Am were investigated in the bloody cockle Anadara senilis exposed via seawater and food in controlled conditions, using animals of different weight groups in order to assess how their bioaccumulation is affected by allometry and, hence, the individual's age. This study is one of the few experiments investigating bioaccumulation capacities of radionuclides in a West-African bivalve. Results showed that allometric relationships were mainly dependent on the exposure pathway considered. Significant relationships with body weight of bloody cockles were found during the uptake from dissolved phase for both radionuclides; they followed inverse power functions: smaller cockles concentrated both radionuclides more than larger ones. In contrast, radionuclide absorption and assimilation efficiencies from water and food, respectively, did not show any significant relationship with weight: only slight variation was observed between small and large organisms for the retention of 241Am accumulated from food. A bioaccumulation model was used to assess the contribution of each pathway of exposure (food vs. water) in organisms grouped in small and large individuals. We found that, regardless of the size, 134Cs was mainly bioaccumulated through the dietary pathway. In the case of 241Am, the relative contribution of each pathway is weight-dependent: major contribution of dissolved pathway in smaller organisms and the major dietary contribution in larger organisms.

A study of the influence of brevetoxin exposure on trace element bioaccumulation in the blue mussel Mytilus edulis.

Marine organisms are exposed to and affected by a multitude of chemicals present in seawater and can accumulate in their tissues a wide range of contaminants as well as natural biotoxins associated with harmful algal blooms (HABs). Trace elements and biotoxins may modify physiological functions in exposed organisms, and studies have been conducted to better understand their respective kinetics and effects in marine species. Despite the increasing concern of concurrent toxic HABs and pollution events due to anthropogenic pressures and global change, very little information is available on their combined effects. Chemical interactions between biotoxins and trace elements have been reported, and exposure to certain biotoxins is known to modify ion transport pathways, suggesting that biotoxins have the potential to alter trace element uptake. Using specific and sensitive radiotracer techniques (radioligand receptor binding assay and γ-spectrometry), this laboratory study examined the influence of pre-exposure to the brevetoxins (PbTxs)-producing microalgae Karenia brevis on the bioaccumulation of selected non-essential (Cd) and essential (Co, Mn and Zn) trace elements in the blue mussel Mytilus edulis. PbTxs are a group of neurotoxins known to accumulate in bivalves but also to have lethal effects on a number of marine organisms including fish and mammals. We found that, over 23 days exposure to the radiotracers, the bioaccumulation of the dissolved essential trace elements Co, Mn and Zn in M. edulis was not significantly affected by pre-exposure to toxic K. brevis. In contrast, the uptake rate constant ku of Cd was significantly higher in the pre-exposed group (p < 0.05), likely caused by a decrease in mussel clearance rates after K. brevis exposure. These results suggest that the effects of algal toxin exposure on bioaccumulation of trace elements in mussels may be trace element-dependent.

Interspecific comparison of radiocesium trophic transfer in two tropical fish species.

The trophic transfer of radiocesium (134Cs) was investigated in two tropical fish, the silver moony Monodactylus argenteus and the spotted scat Scatophagus argus. Juveniles of both species were exposed to dietary 134Cs using the pulse-chase feeding methodology. The food was brine shrimp (Artemia salina) previously exposed to the dissolved radiotracer. Depuration kinetics of 134Cs were followed for 45 d. Results showed that Cs was similarly efficiently assimilated by both species (AE > 50%). The estimated trophic transfer factors in the two species ranked from 1 to 2, suggesting that 134Cs could be biomagnified in both omnivorous species. In complement, dissections of 7 body compartments were carried out at three different times in order to highlight 134Cs organotropism. 134Cs organotropism was similar in both species: more than 50% of 134Cs was quickly distributed in the muscles and skeleton (after 3 days of depuration), which is likely related to the analogous behavior between Cs and K, an essential element for muscle contractions and bone formation.

A double-tracer radioisotope approach to assess simultaneous bioaccumulation of caesium in the olive flounder Paralichthys olivaceus.

To better understand bioaccumulation of radiocaesium in the commercially important Japanese flatfish, Paralichthys olivaceus, the uptake and depuration kinetics of caesium via both seawater and food were assessed simultaneously using controlled aquaria. The pre-conditioned fish were exposed to radionuclides via the two different pathways (aqueous versus dietary) concurrently using two isotopes of caesium, 137Cs and 134Cs, respectively. Dissolved caesium uptake was linear and did not reach a steady state over the course of the 8-day exposure period. Consumption of 134Cs-labelled food led to higher bioaccumulation rates of radioactive Cs than via seawater exposure of 137Cs during uptake and following depuration, though the model-derived long-lived biological half-lives of both pathways was approximately 66 d. Further development of this method for assessing multiple radiocaesium bioaccumulation pathways simultaneously could lead to a promising new approach for studying Cs contamination in marine organisms.

The role of salinity in the trophic transfer of 137Cs in euryhaline fish.

In order to better understand the influence of changing salinity conditions on the trophic transfer of 137Cs in marine fish that live in dynamic coastal environments, its depuration kinetics was investigated in controlled aquaria. The juvenile turbot Scophthalmus maximus was acclimated to three distinct salinity conditions (10, 25 and 38) and then single-fed with compounded pellets that were radiolabelled with 137Cs. At the end of a 21-d depuration period, assimilation efficiencies (i.e. AEs = proportion of 137Cs ingested that is actually assimilated by turbots) were determined from observational data acquired over the three weeks. Our results showed that AEs of 137Cs in the turbots acclimated to the highest salinity condition were significantly lower than for the other conditions (p < 0.05). Osmoregulation likely explains the decreasing AE observed at the highest salinity condition. Indeed, observations indicate that fish depurate ingested 137Cs at a higher rate when they increase ion excretion, needed to counterbalance the elevated salinity. Such data confirm that ambient salinity plays an important role in trophic transfer of 137Cs in some fish species. Implications for such findings extend to seafood safety and climate change impact studies, where the salinity of coastal waters may shift in future years in response to changing weather patterns.

The role of marine biotoxins on the trophic transfer of Mn and Zn in fish.

Essential nutrients are critical for physiological processes of organisms. In fish, they are obtained primarily from the diet, and their transfer and accumulation are known to be impacted by environmental variables such as water temperature, pH and salinity, as well as by diet composition and matrices. Yet, prey items consumed by fish may also contain toxic compounds such as marine toxins associated with harmful algae. These biotoxins have the potential to affect essential trace element assimilation in fish through chemical interactions such as the formation of trace element-toxin complexes or by affecting general fish physiology as in the modification of ion-specific transport pathways. We assessed the influence of dietary exposure to brevetoxins (PbTxs), ichthyotoxic neurotoxins produced by the dinoflagellate Karenia brevis, on trophic transfer of two essential trace elements, Mn and Zn, in a fish model. Using ecologically relevant concentrations of PbTxs and trace elements in controlled laboratory conditions, juvenile turbots Scophthalmus maximus were given food containing PbTxs before or at the same time as a feeding with radiotracers of the chosen essential elements (54Mn and 65Zn). Treatments included simultaneous exposure (PbTxs + 54Mn + 65Zn) in a single-feeding, 3-week daily pre-exposure to dietary PbTx followed by a single feeding with 54Mn and 65Zn, and a control (54Mn and 65Zn only). After a 21-day depuration period, turbot tissue brevetoxin levels were quantified and assimilation efficiencies of 54Mn and 65Zn were assessed. PbTxs were found in turbot tissues in each exposure treatment, demonstrating dietary trophic transfer of these toxins; yet, no differences in assimilation efficiencies of Mn or Zn were found between treatments or the control (p > 0.05). These results indicate that, in our experimental conditions, PbTx exposure does not significantly affect the trophic transfer of Mn and Zn in fish.

Investigations of temperature and pH variations on metal trophic transfer in turbot (Scophthalmus maximus).

Studying dietary metal transfer kinetics is essential to gain a better understanding in global metal accumulation rates and its impacts in marine fish. While there exists a solid understanding on the influence of various biotic factors on this transfer, metal assimilation in fish might be also affected by abiotic factors, as has been observed in marine invertebrates. The present study therefore aims to understand the potential effects of two climate-related master variables, temperature and pH, on the assimilation efficiency (AE) of essential (Co and Zn) and non-essential (Ag) metals in the turbot Scophthalmus maximus using radiotracer tools. Juvenile turbots were acclimated for 8 weeks at two temperatures (17 and 20 °C) and pH (7.5 and 8.0) regimes, under controlled laboratory conditions, and then fed with radiolabelled shrimp (57Co, 65Zn and 110mAg). Assimilation efficiencies of Co and Ag in juvenile turbot, determined after a 21-day depuration period, were not affected by pre-exposition to the different environmental conditions. In contrast, temperature did significantly influence Zn AE (p < 0.05), while pH variations did not affect the assimilation of any of the metals studied. In fact, temperature is known to affect gut physiology, specifically the membrane properties of anterior intestine cells where Zn is adsorbed and assimilated from the ingested food. These results are relevant to accurately assess the influence of abiotic factors in AEs of metals in fish as they are highly element-dependent and also modulated by metabolic processes.

Dietary Zn and the subsequent organotropism in fish: No influence of food quality, frequency of feeding and environmental conditions (pH and temperature).

Trophic transfer of Zn in fish is affected by the type of food and environmental variables such as temperature. However, there is still a lack of knowledge regarding the effects of such factors on Zn organotropism. For this reason, a series of experimental studies have investigated how the distribution and the concentration of Zn is affected by some environmentally-relevant factors (food quality, food availability, water pH, and temperature) in turbot Scophthalmus maximus using radiotracer techniques. In three different experiments, Zn distribution in seven body compartments of juvenile turbot and the calculation of Zn concentration index (IC) for each compartment were compared. Its distribution as well as its concentration in the body compartments of juvenile turbots were not affected by the experimental conditions tested. This apparent consistency in the Zn organotropism can be explained by the ability of the fish to maintain Zn homeostasis at non-toxic Zn concentrations in their diet. These results are important to better understand the trophic transfer of Zn in fish under realistic environmental conditions.

Trophic transfer of essential elements in the clownfish Amphiprion ocellaris in the context of ocean acidification.

Little information exists on the effects of ocean acidification (OA) on the digestive and post-digestive processes in marine fish. Here, we investigated OA impacts (Δ pH = 0.5) on the trophic transfer of select trace elements in the clownfish Amphiprion ocellaris using radiotracer techniques. Assimilation efficiencies of three essential elements (Co, Mn and Zn) as well as their other short-term and long-term kinetic parameters in juvenile clownfish were not affected by this experimental pH change. In complement, their stomach pH during digestion were not affected by the variation in seawater pH. Such observations suggest that OA impacts do not affect element assimilation in these fish. This apparent pCO2 tolerance may imply that clownfish have the ability to self-regulate pH shifts in their digestive tract, or that they can metabolically accommodate such shifts. Such results are important to accurately assess future OA impacts on diverse marine biota, as such impacts are highly species specific, complex, and may be modulated by species-specific metabolic processes.

Comparing single-feeding and multi-feeding approaches for experimentally assessing trophic transfer of metals in fish.

Diet is an important pathway for metal uptake in marine organisms, and assimilation efficiency is one of the most relevant parameters to quantify trophic transfer of metals along aquatic food webs. The most commonly used method to estimate this parameter is pulse-chase feeding using radiolabeled food. This approach is, however, based on several assumptions that are not always tested in an experimental context. The present study aimed to validate the approach by assessing single-feeding and multiple-feeding approaches, using a model species (the turbot Scophthalmus maximus). Using the kinetic data obtained from the single-feeding experiment, the reconstruction of a multi-feeding experiment was tested for consistency with data provided by an actual multi-feeding performed under the same experimental conditions. The results validated the single-feeding approach. Environ Toxicol Chem 2017;36:1227-1234. © 2016 SETAC.

Bioconcentration of Ag, Cd, Co, Mn and Zn in the Mangrove Oyster (Crassostrea gasar) and Preliminary Human Health Risk Assessment: A Radiotracer Study.

Bioaccumulation kinetics of five dissolved metals were determined in the mangrove oyster Crassostrea gasar, using corresponding radiotracers ((54)Mn, (57)Co, (65)Zn, (109)Cd and (110m)Ag). Additionally, their bioaccessibility to human consumers was estimated. Results indicated that over a 14-day exposure (54)Mn and (57)Co were linearly concentrated in oysters whereas (109)Cd, (65)Zn and (110m)Ag were starting to saturate (steady-state not reached). Whole-body concentration factors at 14 days (CF14d in toto) ranged from 187 ± 65 to 629 ± 179 with the lowest bioconcentration capacity for Co and the highest for Ag. Depuration kinetics were best described by a double-exponential model with associated biological half-lives ranging from 26 days (Ag) to almost 8 months (Zn and Cd). Bioaccessible fraction of the studied elements was estimated using in vitro digestions, which suggested that oysters consumed seasoned with lemon enhanced the accessibility of Cd, Mn and Zn to human consumers, but not Ag and Co.

Differential bioaccumulation of (134)Cs in tropical marine organisms and the relative importance of exposure pathways.

Bioaccumulation of (134)Cs was determined in 5 tropical marine species: three bivalves (the oysters Isognomon isognomum and Malleus regula, and the clam Gafrarium pectinatum), one decapod (shrimp Penaeus stylirostris) and one alga (Lobophora variegata). Marine organisms were exposed to the radionuclides via different pathways: seawater (all of them), food (shrimp and bivalves) and sediment (bivalves). Our results indicate that the studied tropical species accumulate Cs similarly than species from temperate regions whereas retention capacities seems to be greater in the tropical species. Bioaccumulation capacities of the two oysters were similar for all the exposure pathways. The alga, and to a lesser extent the shrimp, concentrated dissolved Cs more efficiently than the bivalves (approx. 14 and 7 times higher, respectively). Assimilation efficiencies of Cs in bivalves and shrimp after a single feeding with radiolabelled food were comprised between 7.0 ± 0.4 and 40.7 ± 4.3%, with a variable retention time (half-life -Tb1/2- ranging from 16 ± 3 to 89 ± 55 d). Although the clam lives buried in the sediment, this exposure pathway resulted in low bioaccumulation efficiency for sediment-bound Cs (mean transfer factor: 0.020 ± 0.001) that was lower than the two oyster species, which are not used to live in this media (0.084 ± 0.003 and 0.080 ± 0.005). Nonetheless, Cs accumulated from sediment was similarly absorbed (61.6 ± 9.7 to 79.2 ± 2.3%) and retained (Tb1/2: 37 ± 2 to 58 ± 25 d) for the three bivalves species. Despite the poor transfer efficiency of Cs from food, the use of a global bioaccumulation model indicated that the trophic pathways was the main uptake route of Cs in the bivalves and shrimp. In shelled organisms, shells played a non-negligible role in Cs uptake, and their composition and structure might play a major role in this process. Indeed, most of the Cs taken up from seawater and sediment was principally located on the hard parts of the bivalves and shrimp, with the exception of G. pectinatum, where Cs was mainly distributed in the soft-parts.

Trophic transfer of (110m)Ag in the turbot Scophthalmus maximus through natural prey and compounded feed.

Industrial incidents can result in radionuclide release in the environment, among which (110m)Ag. Indeed, under particular circumstances, non-negligible amounts of (110m)Ag have been measured in the marine environment (as observed in Fukushima Dai-ichi incident). This element can therefore be accumulated by aquatic organisms through different pathways including the trophic transfer. The present study aimed at examining the variation of (110m)Ag assimilation efficiency (AE) by turbots, Scophthalmus maximus, when exposed through different feeds. Pulse-chase feeding experiments were carried out in mesocosms, using radiolabelled feeds (natural prey and commercial pellets). Depuration kinetics of (110m)Ag over 21 days were generally fitted by a two-component exponential model; the ingested radioelement was poorly assimilated by turbots regardless of the food item that was used (AE always <3%). Concentration and subcellular distribution of (110m)Ag in prey did not seem to influence its assimilation by turbot. These results suggest that physiological mechanisms could occur in fish that would prevent the transfer of (110m)Ag from gut lumen to internal organs (e.g. (110m)Ag neutralization in the lumen of the stomach, detoxification mechanisms occurring in the gut).