Sublethal effect of silver and chromium in the green mussel Perna viridis with reference to alterations in oxygen uptake, filtration rate and membrane bound ATPase system as biomarkers

Perna viridis is an ideal animal for studying the impairment caused by the effects of heavy metals that are often detected in coastal areas. Preliminary bioassay tests revealed that the lethal (LC(100)), median lethal (LC(50)) and sublethal (LC(0)) concentration of silver and chromium to P. viridis were 6.5, 4.0, 2.0 mg l(-1) and 4.5, 2.5, 1.0 mg l(-1), respectively. Toxic effect of silver and chromium was evaluated in the green mussel P. viridis, with reference to oxygen consumption, filtration rate and ATPase system in laboratory experiments. These parameters were selected as the end point of sublethal stress. Oxygen consumption and filtration rates were calculated as a measure of decline in the dissolved oxygen level and algal concentration (feed) in the aquaria water, respectively. Silver and chromium affects both oxygen consumption and filtration rate significantly (P<0.01) at 96 h when compared to control. The activity of ATPases system in the gills, hepatopancreas, ovary and muscle of mussels were inhibited by silver and chromium indicating that metals exerted significant toxic effect. The inhibition of Na(+)K(+) ATPase, Ca(2+) ATPase and Mg(2+) ATPase in the mussels were significant (P<0.05) for silver and highly significant (P<0.01) for chromium, which indicates that chromium was more toxic to mussels when compared to silver. The assessment of oxygen consumption, filtration and ATPases system can thus be used as a valid biomarker in aquatic ecotoxicology studies.     

Sublethal Cd-induced cellular damage and metabolic changes in the freshwater crab Sinopotamon henanense

To explore whether sublethal cadmium (Cd) exposure causes branchial cellular damages and affects the metabolic activity in brachyuran crustaceans, the freshwater crab Sinopotamon henanense was exposed to 0.71, 1.43, and 2.86 mg/L Cd²⁺ for 3 weeks. Gill morphology, metabolic activity (activities of isocitrate dehydrogenase (IDH), cytochrome c oxidase (CCO) and lactate dehydrogenase (LDH), mRNA expression of CCO active subunit 1 (cco-1) and ldh, as well as ATP levels) in crab muscle were investigated. The results showed that sublethal Cd exposure caused profound morphological damages in the gills. The branchial epithelial cells were disorganized and vacuolized. Ultrastructurally, a decrease in number of apical microvilli, vacuolized mitochondria, and condensed chromatin were observed in gill epithelial cells. Correspondingly, the Cd exposure also induced downregulations of cco-1 and ldh mRNA expression and reduced activities of IDH, CCO, and LDH, in accordance with the lower ATP level in crab muscle. These results led to the conclusion that gill damage caused by sublethal Cd exposure could lead to an impairment of oxygen uptake of S. henanense, and the inhibition of metabolic activity decreases the oxygen demand of the crab and assists them to survive under the condition of lower oxygen availability. These effects add to our understanding on toxic effects of Cd and survival management of S. henanense subchronically exposed to sublethal Cd.     

Salinity effects on cadmium accumulation in various tissues of the tropical fiddler crab Uca rapax

Pollution of water and sediments by industrial discharges of heavy metals, mainly cadmium (Cd), threatens the biota of mangrove ecotones around the mouth of the Tuy river, in northern-central Venezuela. Short-term lethal Cd concentrations were determined in adult male Uca rapax collected from a clean site and kept in 75% seawater (SW). The 96-h LC50 obtained, 380 micromol (=43 mg) Cd liter(-1), is remarkably higher than values previously reported for other crustacean, mollusc or fish species. Tissue accumulation of Cd was examined in crabs acclimated to 125, 75 or 25% SW and exposed to sublethal CdCl2 levels ranging from 9.0 to 225 micromol liter(-1) (i.e. 1.0-25 mg Cd liter(-1)). The metal contents of carapace, gills, hepatopancreas, muscle and hemolymph rose pronouncedly at the highest concentration tested and particularly in the crabs treated in dilute (=25%) SW. In the latter, analyses of the relative distribution of whole-body Cd burden showed that about 80% is in the carapace, 2.4% in the gills, 4% in hepatopancreas, 3% in muscle, 9.3 in other viscerae, 1% is bound to the dried solids of hemolymph and ca. 0.2% appears to be in solution in this fluid. Exposure of crabs to Cd in dilute SW enhances the rates of entry or uptake of the metal, possibly mediated by Ca2+ transport mechanisms, and may exceed the detoxifying capacity of organs such as the hepatopancreas. Thus, Cd may be more harmful to crab populations experiencing frequent fluctuations of ambient salinity, such as are usual in mangrove swamps.     

Fluoride induced changes in protein metabolism in the tissues of freshwater crab Barytelphusa guerini

Exposure of the freshwater field crab Barytelphusa guerini to a sublethal concentration of NaF (30 mg litre(-1)) caused significant alterations in protein metabolism. After an exposure time of 15 days, the crab was found to have a marked depletion of total protein and free amino acid content. A significant elevation in transaminases, Aspartate aminotransferase (AAT) and Alanine aminotransferase (ALAT), and Glutamate dehydrogenase (GDH) activities was reflected in the free amino acid levels of the tissues.     

Impairments of cadmium on vitellogenin accumulation in the hepatopancreas of freshwater crab <Emphasis Type="Italic">Sinopotamon henanense</Emphasis>

During ovary maturation of crabs, vitellogenin (Vg), a precursor molecule of vitellin (Vn) needed for embryogenesis, can be produced in large quantities in the hepatopancreas and then transported to the ovary by the hemolymph. In the present study, effects of Cd on Vg accumulation in the hepatopancreas and Vg transportation of the freshwater crab Sinopotamon henanense were investigated. We also studied the impacts of Cd on the mRNA expression of genes involved in energy metabolism, protein metabolism, and metallothionein (MT) and glutathione (GSH) synthesis. After Cd treatment, the Vg concentration and the Vg mRNA expression in the hepatopancreas were downregulated. Pearson’s correlation coefficient showed that the Vg level in the hepatopancreas correlated positively with those of the ovary and hemolymph (correlation coefficients 0.844 and 0.749, respectively), suggesting that the Vg transport from the hepatopancreas to the ovary can be impaired by Cd. The levels of carbohydrate and protein in the hepatopancreas of Cd-exposed crabs were decreased, and an inhibited protein metabolism was also observed. Energy production related isocitrate dehydrogenase and cytochrome C oxidase mRNA expressions, and MT and GSH synthesis increased after 10 days of Cd treatment and decreased after 20 days. Cd also caused a time-dependent upregulation of malondialdehyde. Our findings showed that Cd decreased Vg accumulation in the hepatopancreas due to partially excessive energy consumption and an activated defense system in the hepatopancreas, suggesting a possible regulatory mechanism in S. henanense which is the competitive advantage of energy reserves in metabolic Cd stress responses over the high-energy flux during vitellogenesis to ensure a continuous supply of metabolic energy. Moreover, the damage of Vg accumulation in the hepatopancreas caused by Cd could lead to an insufficient accumulation of Vn in the ovary and cause a retardation of oocyte development.     

Organochlorinated contaminants in decapod crustaceans from the coasts of Brittany and Normandy (France)

The contamination and distribution of organochlorinated compounds were considered in three crustacean species (edible crab, Cancer pagurus; spider crab, Maja brachydactyla; velvet swimming crab, Necora puber) from five sites along the coasts of Brittany and Normandy (Western and North-Western France). PCBs (16 single congeners), pp'-DDE and HCB were measured in hepatopancreas, gonads and muscle: in all, 175 samples were analysed. The spider crab was the only species found in the five sampling sites, thus enabling comparison between areas. Specimens from Antifer were much more contaminated (summation operator 16 PCBs in hepatopancreas=2000-4000 ng g(-1) dry weight) than those from other sites (50-1000 ng g(-1) d.w.). Among all the three species, the spider crab appeared more contaminated by PCBs than the edible crab, by a factor 2-3, probably in relation with specific differences in their life cycle. There was no difference due to the gender of the species. Within the different analysed tissues, contamination levels increased from muscle to gonads and hepatopancreas in relation with the fat content. A very similar PCB composition was observed in all samples, PCB fingerprints being characterised by the relative importance of the more persistent PCB congeners: CB153, 138, 180, 187, and 118. Finally, these results were compared to recent food regulations first of maximum marker PCB intake and secondly of maximum dioxin-like PCB intake. By considering the muscle, all samples were far below the regulatory limits; for hepatopancreas and gonads, however, some samples were unfit for human consumption.     

Chronic effects of mercuric chloride on the activities of some enzymes in certain tissues of the fresh water murrel,

Alterations in the activities of some enzymes in the brain, gills, intestine, kidney, liver and muscles have been examined in the fresh water murrel, , after exposure to a sublethal concentration of mercuric chloride (3 μg/1) for 15, 30 and 60 days. The results revealed that after 15 days of exposure amino acid oxidase activity was elevated in brain and liver and inhibited in intestine. The activity of xanthine oxidase was increased in gills, and inhibited in kidney. Thirty days exposure produced significant inhibition in the activities of malate dehydrogenase in liver, glutamate dehydrogenase in gills and brain, aminoacid oxidase in gills, and xanthine oxidase in liver and intestine. In contrast, glutamate dehydrogenase in intestine, kidney and liver and aminoacid oxidase in brain and liver were elevated. After 60 days of treatment, a decrease in the activity of glucose-6-phosphatase was recorded in gills, intestine, kidney and liver. Hexokinase activity in kidney and liver, and malate dehydrogenase in all the six tissues were inhibited. Glutamate dehydrogenase activity in intestine, kidney and liver remained higher than in control fish. In brain, kidney and liver the activity of aminoacid oxidase was elevated, but in gills the enzyme activity decreased. Xanthine oxidase activity was inhibited in intestine and liver.     

Accumulation of organic and inorganic contaminants in shellfish collected in estuarine waters near Pensacola, Florida: contamination profiles and risks to human consumers

We conducted a screening level assessment of contaminants in blue crabs (Callinectes sapidus) and oysters (Crassostrea virginica) from bays and bayous in the Pensacola, FL area. Tissue samples were analyzed for 17 dioxins/furans, 12 dioxin-like PCB (DL-PCBs) congeners, mercury, and various metals. Contaminant levels were compared to screening values (SV) calculated using U.S. EPA recommendations for establishing consumption advisories. All sampling locations exceeded the SV (0.098pgg(-1)) for dioxins/furans/DL-PCBs, based on a Florida-specific consumption rate (46gday(-1)). Arsenic (inorganic), mercury, cadmium, and zinc levels exceeded SVs in samples from select locations, and with the exception of mercury, these locations were generally downstream of known contaminated areas. We also assessed potential human health risks from consumption of these species. Risks to human health were greatest from consumption of crab hepatopancreas, suggesting that consumption of hepatopancreas, whether directly or indirectly, from crabs collected anywhere in the Pensacola Bay region should be avoided.     

Chronic effects of mercuric chloride on the activities of some enzymes in certain tissues of the fresh water murrel, Channapunctatus

Alterations in the activities of some enzymes in the brain, gills, intestine, kidney, liver and muscles have been examined in the fresh water murrel, $\text{Channa}$$\text{punctatus}$, after exposure to a sublethal concentration of mercuric chloride (3 μg/1) for 15, 30 and 60 days. The results revealed that after 15 days of exposure amino acid oxidase activity was elevated in brain and liver and inhibited in intestine. The activity of xanthine oxidase was increased in gills, and inhibited in kidney. Thirty days exposure produced significant inhibition in the activities of malate dehydrogenase in liver, glutamate dehydrogenase in gills and brain, aminoacid oxidase in gills, and xanthine oxidase in liver and intestine. In contrast, glutamate dehydrogenase in intestine, kidney and liver and aminoacid oxidase in brain and liver were elevated. After 60 days of treatment, a decrease in the activity of glucose-6-phosphatase was recorded in gills, intestine, kidney and liver. Hexokinase activity in kidney and liver, and malate dehydrogenase in all the six tissues were inhibited. Glutamate dehydrogenase activity in intestine, kidney and liver remained higher than in control fish. In brain, kidney and liver the activity of aminoacid oxidase was elevated, but in gills the enzyme activity decreased. Xanthine oxidase activity was inhibited in intestine and liver.     

Metal accumulation in the greentail prawn, Metapenaeus bennettae, in Sydney and Port Hacking estuaries, Australia

Metal concentrations of the inshore greentail prawn, Metapenaeus bennettae, and surface sediments from locations within Sydney estuary and Port Hacking (Australia) were assessed for bioaccumulation and contamination. The current study aimed to assess metal concentrations in prawn tissue (tail muscle, exoskeleton, hepatopancreas and gills), relate whole body prawn tissue metal concentrations to sediment metal concentrations and animal size, as well as assess prawn consumption as a risk to human health. Metal concentrations were highest in sediment and prawns from contaminated locations (Iron Cove, Hen and Chicken Bay and Lane Cove) in Sydney estuary compared with the reference estuary (Port Hacking). Concentrations in sediments varied considerably between sites and between metals (As, Cd, Cr, Cu, Ni, Pb and Zn), and although concentrations exceeded Interim Sediment Quality Guideline-Low values, metals (As, Cd, Cr, Cu, Ni, Pb and Zn) were below Australian National Health and Medical Research Council human consumption guidelines in prawn tail muscle tissue. Metal concentrations in prawn tail muscle tissue were significantly different (p?≤?0.05) amongst locations for Pb, Zn and Cd, and metal concentrations were generally highest in gills tissue, followed by the hepatopancreas, exoskeleton and tail muscle. The exoskeleton contained the highest Sr concentration; the hepatopancreas contained the highest As, Cu and Mo concentrations; and the gills contained the highest Al, Cr, Fe and Pb concentrations. Concentrations of Pb, As and Sr were significantly different (p?≤?0.05) between size groups amongst locations.     

Sublethal effects of monocrotophos on locomotor behavior and gill architecture of the mosquito fish, Gambusia affinis

Subacute studies of monocrotophos [Dimethyl (E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] on mosquito fish, Gambusia affinis, were carried out in vivo for 24 days to assess the locomotor behavior, structural integrity of gill, and targeted enzyme acetylcholinesterase (AChE, EC: 3.1.1.7) interactions. Monocrotophos (MCP) can be rated as moderately toxic to G. affinis, with a median lethal concentration (LC(50)) of 20.49 +/- 2.45 mgL(-1). The fish exposed to sublethal concentration of LC(10) (7.74 mgL(-1)) were under stress and altered their locomotor behavior, such as distance traveled per unit time (m min(-1)) and swimming speed (cm sec(-1)) with respect to the length of exposure. Inhibition in the activity of brain AChE and deformities in the primary and secondary lamellae of gill may have resulted in failure of exchange of gases. The maximum inhibition of 95% of AChE activity was observed on days 20 and 24.Morphological aberrations in the gills were also studied during exposure to the sublethal concentration of monocrotophos for a period ranging from 8 to 24 days. The extent of damage in gill was dependent on the duration of exposure. The findings revealed that inhibition in brain AChE activity and structural alteration in gill were responsible for altering the locomotor behavior of exposed fish.     

Effects of cadmium on glutathione synthesis in hepatopancreas of freshwater crab, Sinopotamon yangtsekiense

Cadmium (Cd) is one of the most deleterious heavy metals in aquatic systems that could promote oxidative damage. To explore the effects of Cd exposure of a freshwater crab (Sinopotamon yangtsekiense) on hepatopancreatic glutathione (GSH) synthesis, crabs were exposed to the reagent with a dose range of 7.25-116.00 mg L(-1) for 48 h. The concentrations of GSH, oxidized glutathione (GSSG), NADPH and NADP(+), as well as the activities of enzymes involved in GSH synthesis, i.e. glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamylcysteine synthetase (gamma-GCS) were determined. Progressive depletion of cellular GSH content was observed with the increasing of Cd concentrations, while the level of GSSG remained constant. In response to Cd exposure, crabs showed significant induction of G6PD and NADPH, however, only up to moderate exposures. GR activity remained at a steady level at all exposure concentrations. The activity of gamma-GCS was significantly positively correlated with the Cd concentration. These results suggested that GSH synthesis could be activated against reactive oxygen species induced by lower Cd exposure; under the higher Cd exposure conditions, an inhibition of NADPH-dependant redox cycling and de novo GSH synthesis led to significant decrease in GSH content.     

Effects of Cadmium and Zinc on the feeding behaviour of two freshwater crustaceans: Atyaephyra desmarestii (Decapoda) and Echinogammarus meridionalis (Amphipoda)

Lethal and sublethal effects of Cadmium and Zinc on two freshwater crustaceans were investigated. The 96-h LC50 for Cadmium and Zinc were 42.20 microg l(-1) and 5.43 mg l(-1) for Atyaephyra desmarestii, and 36.17 microg l(-1) and 4.61 mg l(-1) for Echinogammarus meridionalis, respectively. An increase in metal concentrations at sublethal levels resulted in significant reductions of the feeding rate of both species. The LOECs in the chronic feeding assays for Zinc were 1.29 mg l(-1) for A. desmarestii and 0.4 mg l(-1) for E. meridionalis. For Cadmium the LOEC was 6.53 microgl(-1) for both species. The results suggest the potential use of these two crustacean species as test organisms in ecotoxicological tests. Feeding assays appear to be a rapid, cheap and effective tool to be used in biomonitoring studies of Portuguese freshwaters.     

A multi-biomarker approach to assess the impact of farming systems on black tiger shrimp (Penaeus monodon)

This study examined the advantages of the use of biomarkers as an early warning system by applying it to different shrimp farming systems in Soctrang and Camau provinces, main shrimp producers in Mekong River Delta, Vietnam. Shrimp were collected at 15 different farms divided into four different farming systems: three farms were converted from originally rice paddies into intensive shrimp farming systems (IS1, IS2, IS3); three farms were rice-shrimp integrated farming systems (RS4, RS5, RS6); three farms were intensive farming systems (IS7, IS8, IS9); six farms were extensive shrimp farming systems (From ES1 to ES6). Lipid peroxidation (LPO) and total glutathione (GSH) were measured as well as catalase (CAT), glutathione peroxidase (GPX), glutathione S-transferase (GST) and acetylcholinesterase activities (ACHE). Organ specificity was observed between gills and hepatopancreas with generally higher activity of GST in gills (GSTG) whereas the contrary was observed for LPO level in gills (LPOG). Hierarchical clustering and principal component analysis clearly indicated that shrimp reared in extensive culture system formed a distinct group from those reared in intensive or rice-shrimp integrated systems. CAT in gills (CATG), GPX in gills (GPXG) and hepatopancreas (GPXHP) and ACHE in muscle (ACHEM) of shrimp collected in extensive farms showed a general higher level than those in intensively farmed shrimp. On the contrary, we observed clear high levels of GSTG and GST in hepatopancreas (GSTHP) and LPOG and hepatopancreas (LPOHP) of shrimp sampled in intensive and rice-shrimp integrated systems. Thus, we propose that LPO and CAT, GPX, GST and ACHE can be used as a set of biomarkers for the assessment of health condition and can discriminate between shrimp cultivated in different farming systems. These findings provide the usefulness of integrating a set of biomarkers to define the health status of shrimp in different shrimp culture systems.     

Water quality and fish size affect toxicity of endosulfan, an organochlorine pesticide, to rainbow trout

The acute toxicity of endosulfan in juvenile rainbow trout (Oncorhynchus mykiss, 10.61+/-1.69 g) was evaluated in glass aquaria under static conditions. Nominal concentrations of endosulfan in the toxicity test ranged from 1.3 microg l(-1) to 29 microg l(-1). The concentrations of endosulfan that killed 50% of the rainbow trout within 24-h (24-h LC50), 48-h LC50, 72-h LC50, and 96-h LC50 were 19.78, 8.89, 5.28, and 1.75 microg l(-1), respectively. None of the unexposed control fish died, and the first fish died 4 h after exposure to 26.3 microg l(-1) of endosulfan. Survival of fish was significantly increased with increasing fish size and decreased with decreased fish size at the same temperature (p<0.001). Temperature also had a significant effect on survival of fish. Alkalinity at levels above 20 mg l(-1) as CaCO3 significantly increased survival of fish at 19.78 microg l(-1) of endosulfan. Increasing alkalinity from 20 mg l(-1) as CaCO3 to 42 or higher concentrations tested in this study (121 mg l(-1) as CaCO3) significantly increased survival of fish (p<0.01). Total hardness ranging from 55 mg l(-1) as CaCO3 to 126 mg l(-1) as CaCO3 did not affect survival of fish exposed to endosulfan. Endosulfan toxicity was found to be irreversible when fish were exposed to minimum concentrations of endosulfan tested. Histologically, fish gills had lamellar edema, separation of epithelium from lamellae, lamellar fusion, and swelling of the epithelial cells. Melanomacrophage centers were scattered throughout the trunk kidney, head kidney, and spleen. The liver of endosulfan-exposed fish had severe focal necrosis. None of these lesions were seen in unexposed control fish. Results indicate that alkalinity, temperature, and fish size affect endosulfan toxicity of rainbow trout.     

Alteration in some biochemical and enzymological parameters in the snake head fish , exposed chronically to quinalphos

The effect of chronic quinalphos exposure (0.025 mg/1) for 15 and 30 days on the levels of glucose, lactic acid and haemoglobin in the blood; glycogen and lactic acid contents of the liver and muscles; and the activities of hexokinase, lactate dehydrogenase, pyruvate dehydrogenase and succinate dehydrogenase in liver, kidney, intestine, brain, gills and muscles was examined. Blood glucose, lactic acid and haemoglobin levels decreased in quinalphos exposed fish. Glycogen content of liver and muscles increased but lactic acid decreased. Hexokinase was inhibited in intestine and muscles after 30 days of exposure but increase in enzyme activity was noted in gills. Lactate dehydrogenase activity was inhibited in all the six tissues. Pyruvate dehydrogenase activity of liver, kidney, gills and muscles was inhibited. However, in brain the enzyme activity was elevated. Succinate dehydrogenase activity was elevated in intestine and inhibited in other tissues.     

Effects of the anti-fouling herbicide Irgarol 1051 on two life stages of the grass shrimp, Palaemonetes pugio

This study investigated lethal and sublethal effects (glutathione, lipid peroxidation, cholesterol, and acetylcholinesterase) of the anti-fouling herbicide Irgarol 1051 on larval and adult grass shrimp (Palaemonetes pugio). The 96-hour LC50 test for larvae resulted in an estimated LC50 of 1.52 mg/L (95% confidence interval [CI] 1.26-1.85 mg/L). The adult 96-h LC50 was 2.46 mg/L (95% CI = 2.07-2.93 mg/L). Glutathione, lipid peroxidation, cholesterol and acetylcholinesterase levels were not significantly affected in adult grass shrimp by exposure of up to 3.00 mg/L irgarol. Lipid peroxidation and acetylcholinesterase levels in the larvae were significantly higher than controls in the highest irgarol exposures of 1.0 and 2.0 mg/L, respectively. Cholesterol levels were significantly reduced in larvae in all four irgarol concentrations tested while glutathione levels were not significantly affected in larvae. Both lethal and sublethal effects associated with irgarol exposure were only observed at concentrations well above those reported in the environment.     

Fenitrothion, an organophosphate, affects running endurance but not aerobic capacity in fat-tailed dunnarts (Sminthopsis crassicaudata)

We measured aerobic metabolism during cold exposure and exercise performance (run duration and oxygen consumption while running at 1 m s(-1)) in the fat-tailed dunnart Sminthopsis crassicaudata, a dasyurid marsupial, before and after ingestion of 30 mg kg(-1) of fenitrothion, an organophosphate (OP) pesticide. Running endurance of OP-exposed animals was less than half that of control animals over the first 3 days after dosing and 55% of control animal endurance on day 5 post-dose. Despite these declines, peak metabolic rate at this running speed (9.3 times basal metabolic rate; BMR) was unaffected by OP exposure. Peak metabolic rate (PMR) and cumulative oxygen consumption during a 1-h exposure to conditions equivalent to -20 degrees C did not differ between OP-treated and control dunnarts, with PMR averaging 11 times BMR. We conclude that fenitrothion-induced exercise fatigue is not due to limitations in oxygen or substrate delivery to muscle or in their uptake per se, but more likely relates to decreased ability to sustain high-frequency neuromuscular function. The persistence of locomotor impairment following OP exposure in otherwise asymptomatic animals emphasizes the importance of using performance-based measures when characterising sublethal effects of pesticide exposure in an ecological context.     

Alteration in some biochemical and enzymological parameters in the snake head fish Channapunctatus, exposed chronically to quinalphos

The effect of chronic quinalphos exposure (0.025 mg/1) for 15 and 30 days on the levels of glucose, lactic acid and haemoglobin in the blood; glycogen and lactic acid contents of the liver and muscles; and the activities of hexokinase, lactate dehydrogenase, pyruvate dehydrogenase and succinate dehydrogenase in liver, kidney, intestine, brain, gills and muscles was examined. Blood glucose, lactic acid and haemoglobin levels decreased in quinalphos exposed fish. Glycogen content of liver and muscles increased but lactic acid decreased. Hexokinase was inhibited in intestine and muscles after 30 days of exposure but increase in enzyme activity was noted in gills. Lactate dehydrogenase activity was inhibited in all the six tissues. Pyruvate dehydrogenase activity of liver, kidney, gills and muscles was inhibited. However, in brain the enzyme activity was elevated. Succinate dehydrogenase activity was elevated in intestine and inhibited in other tissues.     

Biochemical and locomotor responses of Carcinus maenas exposed to the serotonin reuptake inhibitor fluoxetine

The aim of this study was to assess the effects of the widely used anti-depressant fluoxetine on behaviour (locomotion), moulting, neuromuscular transmission, energy production and anti-oxidant defences’ efficiency of the epibenthic crab Carcinus maenas. Crabs were individually exposed to fluoxetine concentrations for 7 d. Effects on locomotion were assessed at the end of the exposure using an open field test adapted to C. maenas in the present study. Tissue samples were later collected to evaluate fluoxetine effects on physiological functions using the activity of key enzymes and other parameters as biomarkers, namely: N-acetyl-β-glucosaminidase (NAGase) in the epidermis (moulting) and the hepatopancreas; cholinesterases (ChE) in muscle (neuromuscular cholinergic transmission); NADP⁺-dependent isocitrate dehydrogenase (IDH) and lactate dehydrogenease (LDH) in muscle (energy production); glutathione S-transferases (GST) in hepatopancreas (biotransformation and oxidative stress system); glutathione reductase (GR), and glutathione peroxidade (GPx), total glutathione levels (TG) and lipid peroxidation levels in the hepatopancreas (anti-oxidant defences and oxidative damage). Because no information on C. maenas NAGase activity was previously available, its variation during the moult cycle was also investigated. The results showed that locomotion was significantly increased at fluoxetine concentrations equal or above 120 μg L⁻¹, with animals spending more time moving, walking longer distances than controls. Levels of NAGase activity were found to vary in relation to C. maenas moult cycle, but no alterations were observed after exposure to fluoxetine. Significant increases in the activity of ChE, GST and GR enzymes, and the levels of TG were found, with a lowest observed effect concentration (LOEC) of 120 μg L⁻¹. Effects on locomotion were significantly and positively correlated to those induced on ChE activity. The results raise concern when hypothesising conditions of chronic exposure in the wild.