Metallothionein-like Protein Concentrations Research Articles

Partitioning of metals in the tissues and cytosolic fraction of Cerastoderma edule

The concentrations of Cd, Cu and Zn have been determined in the tissues and the cytosolic fraction of the common cockle, Cerastoderma edule, collected from sediments in the Tamar, Plym and Avon estuaries (South West, England). Metal concentrations in the tissues of C. edule from the Avon were lower than those from the Tamar and Plym, except for Cu in the digestive gland. Significant statistical relationships were only obtained between the total sedimentary metal concentrations and Cd in the body of C. edule and Cu in the digestive gland. The cytosolic fraction was extracted from each of the tissues and separated for protein analysis thereby allowing determination of the metal contents in high molecular weight (HMW) compounds, metallothionein-like proteins (MTLP) and very low molecular weight (VLMW) compounds. The digestive glands of C. edule from the Avon had relatively low concentrations of MTLP, whereas MTLP concentrations in the digestive gland of cockles from the Tamar and Plym were higher. The cytosolic fraction of C. edule had relatively low total Cd and Cu concentrations associated with MTLP, whereas Zn was preferentially associated with the HMW and the VLMW components. The results are relevant to metal distributions in C. edule and the role of cytosols in the management of metals by C. edule and other invertebrates.

Monitoring metallothionein-like protein concentrations and cholinesterase activity in tropical cup oysters as biomarkers of exposure to metals and pesticides in the southern Caribbean, Colombia.

Metallothionein-like protein concentrations (MT) and three functionally defined fractions of cholinesterase activity (ChE) were quantified in gill and digestive gland homogenates of tropical cup oysters from 5 nearshore locations in the Colombian Caribbean and correlated with sediment and tissue metal (9 metals) and pesticide (22 organophosphates, OPs, and 20 organochlorines-OCPs), as well as water physical-chemical parameters (salinity, pH, temperature, and dissolved oxygen). Tissue and sediment pesticide concentrations were below detection limits in all samples, whereas sediment and tissue metal concentrations exceeded environmental thresholds at several locations. Tissue MT and ChE biomarkers varied by a factor of 5-6 between locations. Inhibition of cholinesterase activity was negligible for all 5 sites, despite spatial-temporal variation in ChE activity, consistent with below-detection OP concentrations. Tissue MT and ChE biomarkers correlated with tissue and metal sediment concentrations, yet, statistically significant covariance between biomarkers and water chemistry parameters was also observed, indicating that both, metal concentrations and physical-chemical variables, are likely to be responsible for generating the observed spatial-temporal variations in biomarker patterns.

The countryside or the city: Which environment is better for the honeybee?

For a number of years, the decline of honeybee (Apis mellifera) in North America and Europe has been the subject of much debate. Among the many factors proposed by hundreds of studies to explain this phenomenon is the hypothesis that agricultural activities using pesticides contribute to the weakness of bee colonies. Moreover, while urban beekeeping is presently booming in several cities, we do not know if this environment is more beneficial for bees than the typical, rural area. In the summer of 2018, we sampled honeybees (foragers and larvae) in rural (Laurentians) and urban (city of Montreal) areas and compared them using the following biomarkers: carotenoids, retinoids, α-tocopherol, metallothionein-like proteins (MTLPs), lipid peroxidation, triglycerides, acetylcholinesterase activity (AChE) and proteins. Pesticides, pharmaceuticals and personal care products (PPCPs) and metals were also quantified in honeybees' tissues. Our result revealed that, globally, urban foragers had higher levels of insecticides and PPCPs and that metals were in greater concentrations in urban larvae. Compared to rural foragers, urban foragers had higher concentrations of MTLPs, triglycerides, protein and AChE activity. The multifactorial analysis confirmed that insecticides, some metals and PPCPs were the most influential components in the contaminant‒biomarker relationships for both foragers and larvae.

Detecting the effects of chronic metal exposure on benthic systems: Importance of biomarker and endpoint selection

Understanding metal toxicity to benthic systems is still an ecotoxicological priority and, although numerous biomarkers exist, a multi-biomarker and endpoint approach with sediment as the delivery matrix combined with life-history relevant exposure timescales is missing. Here we assess potential toxicity by measuring a suite of biomarkers and endpoints after exposing the ecologically important polychaete Alitta(Nereis)virens to sediment spiked with environmentally relevant concentrations of copper and zinc (and in combination) for 3, 6 and 9 months. We compared biomarker and endpoint sensitivity providing a guide to select the appropriate endpoints for the chosen time frame (exposure period) and concentration (relevant to Sediment Quality Guidelines) needed to identify effects for benthic polychaetes such as A. virens. Target bioavailable sediment and subsequent porewater concentrations reflect the global contamination range, whilst tissue concentrations, although elevated, were comparable with other polychaetes. Survival reduced as concentrations increased, but growth was not significantly different between treatments. Metabolic changes were restricted to significant reductions in protein after 9 months exposure across all copper concentrations, and reductions in lipid at high copper concentrations (3 months). Significant changes in feeding behaviour and increases in metallothionein-like protein concentration were limited to the medium and high copper and zinc concentrations, respectively, both after 6 months exposure. Despite data highlighting A. virens′ metal tolerance, DNA damage and protein concentrations are the most sensitive biomarkers. Copper and zinc cause biomarker responses at concentrations routinely found in coastal sediments that are characterised as low contamination, suggesting a reappraisal of the current input sources (especially copper) is required.

Selection of biochemical and physiological parameters in the croaker Micropogonias furnieri as biomarkers of chemical contamination in estuaries using a generalized additive model (GAM)

Biochemical and physiological parameters in the croaker Micropogonias furnieri were evaluated as biomarkers of chemical contaminants in estuaries. Juvenile croakers (10–20 cm total body length) were collected in summer and winter (2011 and 2012), in two sites at the Lagoa dos Patos estuary (southern Brazil). Fish were also collected in summer (2011 and 2012) in one site at the Barra do Chui estuary (southern Brazil). Tissue (gills, muscle or liver) samples were dissected and analyzed for contaminants [metals, polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCPs)] and selected biochemical and physiological parameters [metallothionein-like proteins (MTLP), acetylcholinesterase (AChE) activity, ethoxyresorufin-O-deethylase (EROD) activity, and lipid peroxidation (LPO)]. Additionally, water samples were collected for water chemical analyses (salinity, pH, temperature, dissolved oxygen, total alkalinity, and sulfate concentration). Data obtained were integrated and analyzed, using Principal Component Analysis (PCA) and Generalized Additive Model (GAM) approaches. Results showed that changes in concentrations of chemical contaminants and responses of biochemical and physiological parameters did not show any pattern according to the site, season and year of fish collection. However, they were influenced by fish body length and water temperature and salinity. Liver LPO and EROD activity were not responsive to PAHs. However, liver LPO was responsive to HCB, p,p-DDD, p,p-DDT and endosulfan sulfate. In turn, gill MTLP concentration, muscle AChE activity and liver EROD activity were responsive to non-essential metals (Pb, Cd and Ag). Considering that the ecotoxicological modeling approach adopted (GAM) accounted for biological, spatial and temporal variability of data associated with fish body size and site, season and year of fish collection, gill MTLP concentration, muscle AChE activity, as well as liver LPO and EROD activity can be considered as reliable biomarkers of fish exposure to chemical contaminants in estuaries.

Biomarkers of waterborne copper exposure in the Neotropical fish Prochilodus lineatus

The main goal of the present study was to investigate the effects of acute exposure to copper (Cu) using a Neotropical freshwater fish as sentinel species through multi biomarkers analysis at different biological levels. Juveniles of Prochilodus lineatus were kept under control condition (no Cu addition in the water) or exposed to environmentally relevant concentrations of waterborne Cu (5, 9 and 20μgL−1) for 96h. These concentrations were selected to bracket the current Brazilian water quality criteria for Cu in fresh water (9 and 13μgL−1 dissolved copper). Endpoints analyzed included ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, reduced glutathione (GSH) and metallothionein-like protein (MT) concentration, lipid peroxidation (LPO) level, tissue damage index, and incidence of free melano-macrophages (FMM) and melano-macrophage centers (MMC) in the liver. They also included DNA damage (frequency of nucleoids per comet class, number of damaged nucleoids per fish and DNA damage score) in erythrocytes, as well as muscle and brain acetylcholinesterase (AChE) activity and behavioral parameters (swimming distance and velocity, time spent swimming and swimming activity in the upper and lower layers of the water column). Fish exposed to any of the Cu concentrations tested showed increased liver MT concentration and LPO level, higher number of damaged nucleoids in erythrocytes per fish, and inhibited muscle AChE activity. Also, increased liver SOD activity was observed in fish exposed to 9 and 20μgL−1 Cu. Fish exposed to 5 and 9μgL−1 Cu spent lower amount of time swimming. Fish exposed to 9μgL−1 Cu showed increased swimming distance and velocity while those exposed to 20μgL−1 Cu had lower swimming distance and velocity, as well as, spent less time swimming in the lower layer of the water column when compared to those kept under control condition. These findings indicate that Cu exposure at environmentally relevant concentrations (below or close to the current Brazilian water quality criteria) induced significant biological (histological, biochemical and genetic) and ecological (swimming and exploratory abilities) damages in the Neotropical fish P. lineatus. They also suggest that MT concentration, DNA damage (comet assay), LPO (TBARS method), SOD and AChE activity, together with swimming behavior analyses are potential biomarkers to assess and monitor areas impacted by Cu in fresh water.

Biomarkers of waterborne copper exposure in the guppy Poecilia vivipara acclimated to salt water

The responses of a large suite of biochemical and genetic parameters were evaluated in tissues (liver, gills, muscle and erythrocytes) of the estuarine guppy Poecilia vivipara exposed to waterborne copper in salt water (salinity 24ppt). Activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase, and glutathione S-transferase), metallothionein-like protein concentration, reactive oxygen species (ROS) content, antioxidant capacity against peroxyl radicals (ACAP), and lipid peroxidation (LPO) were evaluated in liver, gills, and muscle. Comet assay score and nuclear abnormalities and micronucleated cell frequency were analyzed in peripheral erythrocytes. The responses of these parameters were evaluated in fish exposed (96h) to environmentally relevant copper concentrations (5, 9 and 20μgL−1). In control and copper-exposed fish, no mortality was observed over the experimental period. Almost all biochemical and genetic parameters proved to be affected by waterborne copper exposure. However, the response of catalase activity in liver, ROS, ACAP and LPO in muscle, gills and liver, and DNA damages in erythrocytes clearly showed to be dependent on copper concentration in salt water. Therefore, the use of these parameters could be of relevance in the scope of biomonitoring programs in salt water environments contaminated with copper.

Evaluation of the potential impact of polluted sediments using Manila clam Ruditapes philippinarum: bioaccumulation and biomarker responses

An assessment was made to monitor the short-term impact of heavily polluted sediments that may move out from the brackish man-made Lake Shihwa outside of the sea dike due to operations of a tidal power plant. Here, we exposed the Manila clam Ruditapes philippinarum collected from the western coast of Korea to natural sediment under lab condition for 96h. Sediments were collected from Lake Shihwa and outside of the sea dike representing polluted and reference conditions, respectively. The results of chemical analysis revealed that the concentrations of nonylphenol and heavy metals in water and sediment from the inner region of Lake Shihwa were significantly higher than those of reference sediments. After 48 and 96h of exposure, 30 specimens of clams were sampled from each experimental condition, and concentrations of nonylphenol and metals were measured in clams, water, and sediments. Several biomarkers, including concentrations of metallothionein-like proteins, and activities of the antioxidant enzymes glutathione S-transferase and catalase were determined in clams to characterize the effects of polluted sediments to clams. After 96h of exposure, R. philippinarum assimilated nonylphenol up to 71 times compared to initial concentrations. However, there was no apparent uptake of heavy metals into the clams. Additionally, antioxidant enzymes exhibited higher activities in clams exposed to the polluted sediment. The results of the present study with physiological responses in R. philippinarum suggest that sediment transportation caused by the operation of a tidal power plant in Lake Shihwa will have striking effects on benthic organisms in the adjacent coastal area.

Copper and zinc detoxification in Gammarus pulex (L.)

To negate the toxicity of labile intracellular metals, some aquatic organisms partition metals into specific subcellular locations for detoxification, namely the soluble heat-stable cytosol and insoluble metal-rich granules. The aim of the present study was to characterise these subcellular storage sites in the freshwater crustacean Gammarus pulex (Linnaeus) following in situ exposures upstream (Drym, low metal) and downstream (Relubbus, elevated metal) of copper- and zinc-rich inflows into the River Hayle (Cornwall, UK). In the cytosol of gammarids exposed at Relubbus, copper and zinc associated to a 7.5-kDa metallothionein-like protein (MTLP) that was largely absent from gammarids prior to exposure. Exposure at Relubbus caused MTLP concentrations to increase 4- to 5-fold between days 2 and 4, indicating an induction response to increased labile intracellular metal. On day 16, spherical calcium-rich granules (0.5-2.5 μmol l(-1)) were visualised and analysed in the posterior caeca of gammarids exposed at both sites. Following exposure at Relubbus, granules contained trace amounts of copper, but zinc was absent. Granules in gammarids exposed at Drym contained no detectable copper or zinc. Granule formation appeared to be independent of exposure. Within the posterior caeca, granules have been associated with calcium storage during the crustacean molt, rather than in detoxification of trace metals. However, the granular copper burden appeared to follow environmental Cu availabilities. Thus, we describe Cu sequestration within molt-cycle calcium storage granules. As both MTLP concentrations and granule formation in crustaceans are affected upon by molting, we hypothesise that detoxification might impact upon this existing process.

ASSESSMENT OF WATER QUALITY IN COASTAL WATERS OF FERNANDO DE NORONHA, BRAZIL: BIOMARKER ANALYSES IN AMPHISTEGINA LESSONII

Research Article| January 01, 2012 ASSESSMENT OF WATER QUALITY IN COASTAL WATERS OF FERNANDO DE NORONHA, BRAZIL: BIOMARKER ANALYSES IN AMPHISTEGINA LESSONII Martina de Freitas Prazeres; Martina de Freitas Prazeres Instituto de Ciências Biológicas, Universidade Federal do Rio Grande – FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, Rio Grande do Sul, Brazil. Caixa Postal 474 Search for other works by this author on: GSW Google Scholar Samantha Eslava Martins; Samantha Eslava Martins Instituto de Ciências Biológicas, Universidade Federal do Rio Grande – FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, Rio Grande do Sul, Brazil. Caixa Postal 474 Search for other works by this author on: GSW Google Scholar Adalto Bianchini Adalto Bianchini * Instituto de Ciências Biológicas, Universidade Federal do Rio Grande – FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, Rio Grande do Sul, Brazil. Caixa Postal 474 *Correspondence author: E-mail: adaltobianchini@furg.br Search for other works by this author on: GSW Google Scholar Journal of Foraminiferal Research (2012) 42 (1): 56–65. https://doi.org/10.2113/gsjfr.42.1.56 Article history received: 11 Jan 2011 accepted: 08 Aug 2011 first online: 03 Mar 2017 Cite View This Citation Add to Citation Manager Share Icon Share MailTo Twitter LinkedIn Tools Icon Tools Get Permissions Search Site Citation Martina de Freitas Prazeres, Samantha Eslava Martins, Adalto Bianchini; ASSESSMENT OF WATER QUALITY IN COASTAL WATERS OF FERNANDO DE NORONHA, BRAZIL: BIOMARKER ANALYSES IN AMPHISTEGINA LESSONII. Journal of Foraminiferal Research 2012;; 42 (1): 56–65. doi: https://doi.org/10.2113/gsjfr.42.1.56 Download citation file: Ris (Zotero) Refmanager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentBy SocietyJournal of Foraminiferal Research Search Advanced Search Abstract Symbiont-bearing foraminiferal assemblages from reef environments have proven useful as ecological indicators for water quality. Studies with other organisms have utilized cellular biomarkers to elucidate mechanisms of stress response. Our goal was to combine these approaches, focusing on populations of the symbiont-bearing foraminifer Amphistegina lessonii from Fernando de Noronha Archipelago (northeastern Brazil). Reef-rubble samples containing A. lessonii were collected at five sampling sites on the leeward side of the main island. Three sites were located inside and two outside the Fernando de Noronha National Marine Park (FNNMP) area. Foraminifers were evaluated regarding their density, degree of bleaching, antioxidant capacity against peroxyl radicals (ACAP), lipid peroxidation (LPO), protein carbonylation (PC), and concentrations of metallothionein-like proteins (MTs). Densities of A. lessonii were lower at sites located outside the FNNMP, compared with those inside the FNNMP, where the maximum average was 159±29 individuals/100 cm2. In individuals from sites located outside FNNMP, bleaching frequency reached 25%, though <2%; at FNNMP sites (<1.8%;). ACAP was higher (two-fold) in individuals collected at the FNNMP sites, which showed lower LPO and PC levels. MTs concentration did not differ significantly among sites. Zinc, copper, and lead concentrations were very high at stations located outside FNNMP. Dissolved organic carbon concentration was high at all sampling sites. Redundancy analysis revealed a strong correlation between biomarkers and water quality among sampling sites. These results show that the biomarker approach would be effective in identifying impacted areas and underlying causes, providing a reliable diagnostic for environment health. You do not have access to this content, please speak to your institutional administrator if you feel you should have access.

Biomarkers response to zinc exposure in the symbiont-bearing foraminifer Amphistegina lessonii (Amphisteginidae, Foraminifera)

The acute effects of zinc (Zn) were evaluated in the symbiont-bearing foraminifer Amphistegina lessonii from the Fernando de Noronha Archipelago (Northeastern Brazil). Foraminifers were acutely (48h) exposed to dissolved Zn concentrations ranging from 9.5 to 93.4μg Zn/l. Endpoints analyzed included mortality, visual alterations (white spots and dark-brown areas in the test), oxidative stress biomarkers (reactive oxygen species generation, lipid peroxidation and total superoxide dismutase activity), and concentration of metallothionein-like proteins in whole individuals after Zn exposure. No significant mortality was observed during the 48-h exposure period to waterborne Zn. However, a significant percentage of individuals showed visual alterations (white spots and/or dark-brown areas in the test) after 24 and 48h of Zn exposure. In fact, a significant positive correlation between this endpoint and dissolved Zn concentrations was observed for both times of exposure. Based on this endpoint, the 24-h and 48-h EC50 values and their corresponding 95% confidence intervals for total measured Zn concentrations were calculated as 112.2 (86.5–199.5) and 43.6 (34.9–57.3) μg Zn/l, respectively. Based on the dissolved Zn concentrations, they were 100.7 (75.3–175.9) and 38.2 (29.7–49.4) μg Zn/l, respectively. Therefore, a significant increase in Zn toxicity was observed with increasing time of exposure. After 48h of Zn exposure, whole body antioxidant capacity was lower in normal-appearing individuals than in those at the initial stage of bleaching. Increases in lipid peroxidation, metallothionein-like protein concentration and total SOD activity was observed at a greater extent in pale/partly-bleached individuals associated with an increased Zn toxicity measured as visual alterations. These findings suggest that an activation of some components of the antioxidant system occurred in A. lessonii to counteract the oxidative stress induced by Zn exposure, and consequently avoid a possible complete loss of the symbiont.

Mechanisms of waterborne Cu toxicity to the pond snail Lymnaea stagnalis: Physiology and Cu bioavailability

We examined the mechanisms of toxicity of waterborne Cu to the freshwater pond snail Lymnaea stagnalis. The snail is one of the most sensitive species to acute Cu exposure (96 h LC 50, LC 20: 24.9, 18.0 μg l −1); they are not protected by the water quality criteria of the US EPA. Tissue Na and Ca were also reduced by Cu in the acute exposure. In contrast, during 28 d chronic exposures to Cu in the presence of food, which resulted in higher DOC concentrations, there was no significant mortality but an inhibition of growth, which may reflect a re-allocation of resources to detoxification. Cu detoxification was evidenced in chronic exposure by increases in metallothionein-like protein concentrations and Cu binding to metal-rich granules, decreases in thiobarbituric acid-reactive substances, and changes in the subcellular distribution in the soft tissues. Our results demonstrated that apart from external Cu bioavailability, compartmentalization of metals within the cells can alter toxicity of Cu to the snails.

Variation of metallothionein-like protein and metal concentrations during the reproductive cycle of the cockle Cerastoderma glaucum from an uncontaminated site: A 1-year study in the Gulf of Gabès area (Tunisia)

Natural variation of metallothionein-like protein (MTLP) concentrations in the cockle Cerastoderma glaucum were analysed over a 1-year period in specimens collected from a relatively uncontaminated site in the Gulf of Gabès (Tunisia). Sampling was carried out from June 2004 to June 2005. At each sampling time, water temperature and salinity were measured. Levels of zinc, cadmium and copper were measured in the whole soft tissues of the cockle C. glaucum. MT-like protein and subcellular metal partitioning were measured in the digestive gland. A histological study was conducted simultaneously to follow the development of the gonad in relation to the reproductive cycle of this bivalve. Results showed a significant fluctuation in MTLP concentrations throughout the whole year, with a maximum value measured in summer and in the late winter. The relationship between MTLP and metal concentrations, abiotic (temperature and salinity) and biotic (mass of target tissues) and also maturity index (MI) factors were examined. Metal levels in the tissues are likely to change during the season, reflecting variability in metabolism and/or a change in tissue weight. However, possible seasonal changes in trace metal concentration are unlikely to entirely explain the observed pattern in the MTLP levels. The physiological changes caused by gonad development and food abundance (change in tissue mass) contribute to a significant variability in MTLP levels in the cockle C. glaucum and can interfere with the estimation of level changes induced by anthropogenic sources.

Differential roles of metallothionein‐like proteins in cadmium uptake and elimination by the scallop Chlamys nobilis

Metallothionein-like proteins (MTLPs) are important inducible ligands for Cd detoxification, but whether MTLPs are involved in Cd bioaccumulation remains unclear. The present study investigated the potential interrelationships between MTLP turnover and Cd biokinetics in the scallop Chlamys nobilis with or without dissolved Cd pre-exposure. After Cd pre-exposure, increased synthesis rate and standing stock concentration of MTLPs as well as increased dissolved Cd uptake rate, were found in C. nobilis. However, the level of newly sequestrated Cd by MTLPs in the pre-exposed scallops was similar to that of the controls, indicating that MTLPs had no effect on Cd uptake from the dissolved phase. Instead, Cd uptake rate was positively related to the Cd binding rate of cellular debris and organelles. In addition, no significant difference was observed in the Cd dietary assimilation efficiency of the scallops, showing that MTLPs played a minimal role in Cd uptake from the dietary phase. Conversely, the Cd efflux rate constant decreased by 68 to 75% in the pre-exposed scallops in comparison with that in the control. Correspondingly, progressive elevations of MTLP-bound Cd and standing stock MTLP concentration were observed in the pre-exposed individuals during the Cd elimination period. In the pre-exposed scallops, the faster de novo MTLP synthesis could remobilize and recycle more Cd from other intracellular pools, effectively reducing the Cd elimination rate. The present study thus showed the differential roles of MTLPs in Cd uptake and elimination in the bivalves.

Metallothionein-like proteins in the blue crab Callinectes sapidus: Effect of water salinity and ions

The effect of water salinity and ions on metallothionein-like proteins (MTLP) concentration was evaluated in the blue crab Callinectes sapidus. MTLP concentration was measured in tissues (hepatopancreas and gills) of crabs acclimated to salinity 30 ppt and abruptly subjected to a hypo-osmotic shock (salinity 2 ppt). It was also measured in isolated gills (anterior and posterior) of crabs acclimated to salinity 30 ppt. Gills were perfused with and incubated in an isosmotic saline solution (ISS) or perfused with ISS and incubated in a hypo-osmotic saline solution (HSS). The effect of each single water ion on gill MTLP concentration was also analyzed in isolated and perfused gills through experiments of ion substitution in the incubation medium. In vivo, MTLP concentration was higher in hepatopancreas than in gills, being not affected by the hypo-osmotic shock. However, MTLP concentration in posterior and anterior gills significantly increased after 2 and 24 h of hypo-osmotic shock, respectively. In vitro, it was also increased when anterior and posterior gills were perfused with ISS and incubated in HSS. In isolated and perfused posterior gills, MTLP concentration was inversely correlated with the calcium concentration in the ISS used to incubate gills. Together, these findings indicate that an increased gill MTLP concentration in low salinity is an adaptive response of the blue crab C. sapidus to the hypo-osmotic stress. This response is mediated, at least in part, by the calcium concentration in the gill bath medium. The data also suggest that the trigger for this increase is purely branchial and not systemic.

Decoupling of cadmium biokinetics and metallothionein turnover in a marine polychaete after metal exposure

This study investigated the kinetics of Cd bioaccumulation, detoxification, subcellular distribution, and efflux in the nereid polychaete Perinereis aibuhitensis after Cd pre-exposure. Cd pre-exposure increased the Cd body burden in the worms, but did not affect the overall Cd uptake and efflux rates and metallothionein-like protein (MTLP) concentrations. During short-term exposure to dissolved Cd, Cd in the cytosolic fraction increased after Cd pre-exposure, and this fraction also increased during the Cd efflux period, indicating that the insoluble fraction of Cd was presumably lost at a faster rate than the loss of cytosolic Cd. Even though the MTLP concentration remained comparable after Cd pre-exposure, both the MTLP synthesis rate and the degradation rate increased, thus leading to a high MTLP turnover in the Cd-exposed worms. However, Cd uptake and efflux into different protein size fractions did not follow the patterns of MTLP synthesis and degradation, strongly suggesting that Cd kinetics is decoupled from the MTLP kinetics in the worms. Our study adds to an increasing body of evidence on the complicated relationship between metal biokinetics and MTLP kinetics in different groups of marine invertebrates which have strong contrasts in their metal handling strategies.

The effect of in situ cadmium contamination on some biomarkers in Cerastoderma glaucum

The present study was designed to detect in situ effects of cadmium on marine organisms Cerastoderma glaucum from the gulf of Gabès (Tunisia). Six sampling stations were chosen: one site, relatively far from known local source of pollution, was considered as uncontaminated site and five sites clearly exposed to anthropogenic impact. Metallothionein like protein (MTLP) and sub-cellular metal partitioning were measured in cockles (C. glaucum) gills and digestive gland. Various biomarkers were also measured, including malondialdehyde (MDA) in the digestive gland and acetylcholinesterase activity (AChE) in the remainder. The health status of the cockles was assessed by using the condition index (CI). Significant differences between sites were noted for Cd concentrations, (CI) and also for the three studied biomarkers. Significant higher biomarkers response was measured in cockles from stations located in the northern part of the gulf, which are exposed directly to industrial and urban effluents, whereas the response of most biomarkers was minimal at the reference station. Positive and significant correlations were observed between MTLP and Cd concentrations in the digestive gland and in the gills. However, it must be noted that these correlations were more significant in the digestive gland, suggesting that compared to the gills, the digestive gland of C. glaucum is more suitable for monitoring metal pollution. The subcellular distribution of Cd showed that the soluble fraction was the major compartment for Cd storage, a pattern which is due to the role of MTLP in Cd detoxication. But at the most contaminated site (EH), cadmium in the digestive gland was preferentially accumulated in the insoluble fraction (P1) suggesting that the MTLP capacity in binding metals was not sufficient to avoid the binding of Cd to the insoluble fraction. Furthermore, the MTLP concentrations in the cockles from this site are lower than expected. So in highly polluted sites, MTLP in C. glaucum should not be used as a useful biomarker for metal pollution. The lipid peroxidation as presented by malondialdehyde levels, and MT-like protein concentrations increased in cockles exposed to cadmium contamination. We can, therefore, hypothesize that Cd could induce MTLP synthesis and MDA increase. While AChE had distinct and specific pattern showing that cadmium is not the only factor of the inhibition of cholinesterase activity. There are other polluting inputs engendering this inhibition.

Metallothionein-like Protein and Tissue Metal Concentrations in Invertebrates (Oligochaetes and Chironomids) Collected from Reference and Metal Contaminated Field Sediments

The relationships between metallothionein-like protein (MTLP) and tissue metal (Ni, Cd, Cu, Zn, Pb) concentrations were examined in Oligochaetes and Chironomid larvae collected from reference and metal contaminated sediments. Tissue metal concentrations, for example, Cd reaching 0.3 μmol/g in oligochaetes and Cu, reaching 0.8 μmol/g in chironomids collected from Great Lakes reference sites (Environment Canada) indicated the presence of bioavailable metal at some sites. The concentration of MTLP ranged from 3.3 (± 1.0) to 11.5 (± 4.5) nmol MTLP/g in oligochaetes, and 2.7 (± 1.0) to 8.0 (± 2.7) nmol MTLP/g in chironomids. Individually, tissue metal concentrations were relatively poor (r 2 = 0.002–0.52) predictors of MTLP, but when multiple metals were used the relationship became stronger (oligochaetes r 2 = 0.84, p < 0.001; chironomids r 2 = 0.61, p < 0.001), suggesting that MTLP was induced by multiple metals. Tissue metal and MTLP concentrations in invertebrates from the metal contaminated slips of Collingwood Harbour were not elevated above reference levels, indicating that only a fraction of the metal in the sediment is bioavailable. Chironomids collected from lakes contaminated by atmospheric fallout of smelting emissions (Rouyn-Noranda, QC) had elevated tissue metal concentrations but MTLP concentration did not consistently reflect elevated tissue metal. Overall, MTLP concentrations in invertebrates collected from metal-contaminated sites were not significantly higher than concentrations in reference site-collected invertebrates. The data presented here suggest that, at some sites, MTLP concentration in field-collected organisms would be a useful surrogate for tissue metal but, because of a lack of detailed taxonomy (organisms were designated as chironomids or oligochaetes) and the relatively small sample sizes of this study, no definitive conclusion regarding the suitability of MTLP concentration as a field-based biomarker of metal exposure can be made without further study

Multigenerational acclimation of Daphnia magna to mercury: Relationships between biokinetics and toxicity

We examined the effects of multigenerational exposure of mercury (Hg) on Hg toxicity and biokinetics in a population of Daphnia magna. After chronic Hg exposure at 3.8 microg Hg/L, the first generation (F0) adults had an elevated 24-h median lethal concentration (LC50) of Hg (76 microg/L) when compared to the control adults (56 microg/L). The dissolved influx rate of Hg was depressed significantly in the Hg-treated adults, which was accompanied by a reduced ingestion rate and enhanced induction of metallothionein-like proteins (MTLP). The second-generation (F1) juveniles originating from the control and exposed lines had no major differences in these parameters (except the dietary assimilation efficiency). Recovery from Hg stress enhanced the vulnerability of F1 adults to Hg toxicity, with a reduced 48-h LC50 (44 microg/L) and a decreased concentration of MTLP (80% of control). Nevertheless, Hg-treated F1 adults had similar tolerance (in terms of LC50s) as the control line, indicating that D. magna acclimated to Hg stress after the first generation of exposure. No major difference occurred in the Hg biokinetics and toxicity among different groups of F2 daphnids. However, the F2 neonates produced by the Hg-treated F1 adults had much higher 48-h LC50 (149 microg/L) and MTLP concentration (148% of control) when there was continuous Hg exposure after birth. We concluded that acclimation to Hg stress occurred quickly in D. magna, though animals recovering from Hg stress were more vulnerable to Hg toxicity. Both ingestion rate and MTLP may not be good biomarkers of Hg stress in the field, because acclimation can be achieved through multigenerational exposure to elevated Hg concentrations.

Effects of buthionine sulfoximine nifurtimox and benznidazole upon trypanothione and metallothionein proteins in Trypanosoma cruzi.

Proteins rich in sulfhydryl groups, such as metallothionein, are present in several strains of the parasite Trypanosoma cruzi, the etiological agent of Chagas' disease. Metallothionein-like protein concentrations ranged from 5.1 to 13.2 pmol/mg protein depending on the parasite strain and growth phase. Nifurtimox and benznidazole, used in the treatment of Chagas' disease, decreased metallothionein activity by approximately 70%. T. cruzi metallothionein was induced by ZnCl2. Metallothionein from T. cruzi was partially purified and its monobromobimane derivative showed a molecular weight of approximately 10,000 Da by SDS-PAGE analysis. The concentration of trypanothione, the major glutathione conjugate in T. cruzi, ranged from 3.8 to 10.8 nmol/mg protein, depending on the culture phase. The addition of buthionine sulfoximine to the protozoal culture considerably reduced the concentration of trypanothione and had no effect upon the metallothionein concentration. The possible contribution of metallothionein-like proteins to drug resistance in T. cruzi is discussed.