Effect of heavy metals (Cu,Cd and Pb) on aspartate and alanine aminotransferase in Ruditapes philippinarum (Mollusca: Bivalvia)

The accumulation of cadmium, copper and lead and their effects on aspartate and alanine aminotransferases in digestive gland, gills, foot and soft body in the clam Ruditapes philippinarum were examined. The animals were exposed to different concentrations: Cd (200–600 μg·l⁻¹), Pb (350–700 μg·l⁻¹) and Cu (10–20 μg·l⁻¹) for 7 days. The highest concentrations were found in digestive gland for cadmium and copper, and in gills for lead, and the lowest values were observed in the foot. Aspartate aminotransferase activity (AST), in general, was not inhibited by cadmium, lead or copper during the exposure. Only in clams exposed to cadmium (600 μg·l⁻¹, 7 days) and copper (20 μg·l⁻¹, 5 days) were observed significant differences (P<0.05) in foot and gills, respectively, with respect to control. In the case of alanine aminotransferase activity (ALT), significant differences were observed for cadmium and lead in treated animals with respect to control. With regard to copper, a decrease in ALT was observed in gills and foot exposed to 20 μg·l⁻¹. A significant correlation (P<0.05) was observed between ALT and metal accumulation for cadmium, copper and lead in gills. In the case of soft body, only cadmium and lead showed a significant correlation. In summary, R. philippinarum can be considered a bioindicator species for cadmium and lead accumulation and ALT could be useful as biomarker of sublethal stress for these metals in soft tissues and gills. Only gills can be considered an adequate target tissue for copper.     

Biomarkers of toxicity in Clarias gariepinus exposed to sublethal concentrations of polycyclic aromatic hydrocarbons

Physiological, biochemical and histological indices in Clarias gariepinus broodstock, and teratogenic indices in embryos exposed to sublethal concentrations of naphthalene, phenanthrene and pyrene were investigated in 2014 using a static-renewal bioassay protocol. Phenanthrene (1.41 mg l^?1) was the most toxic, followed by pyrene (1.53 mg l^?1) and naphthalene (7.21 mg l^?1), based on 96 h LC50 values. Hepatosomatic indices were significantly higher in naphthalene- and pyrene-treated males compared with solvent controls, whereas fecundity in females was significantly lower by factors of 2.4 (naphthalene), 2.8 (phenanthrene) and 2.4 (pyrene), compared with controls. Catalase levels were lower in female phenanthrene-treated fish compared with controls. Histological alterations observed in PAH-treated fish include oedema, inflammatory cells, epithelial lifting and hyperplasia in the gills, vacuolation, haemosiderin pigments and sinusoidal congestion in the liver, and degenerated zona radiata in the ovary. Teratogenic effects were not observed, as evidenced by the lack of histological alterations in embryos spawned from pre-exposed broodstock. Sex-specific responses and the utility of biomarkers at cellular and individual levels of organisation are therefore demonstrated for holistic evaluations of polycyclic aromatic hydrocarbons in ecotoxicological studies.     

Effect of exposure to cadmium on the tropical freshwater prawn Macrobrachium rosenbergii

The effect of cadmium (Cd) on mortality, resistance and bioaccumulation in the tropical freshwater prawn Macrobrachium rosenbergii from Egypt were studied. Survival of prawns exposed to Cd doses over 60 μg l^?1 were significantly lower than that of prawns exposed to lower doses. After 96 h prawns exposed to >40 μg l^?1 of Cd had a greater reduction in total haemocyte count and phagocytic activity than those exposed to lower concentrations. Bioaccumulation of Cd in the gills, hepatopancreas and muscles was variable. Cadmium accumulated significantly in the gills and hepatopancreas, but Cd accumulation in the muscles increased only marginally. Macrobrachium rosenbergii manifested histopathological alterations in the gills, hepatopancreas and muscles when exposed to various concentrations of cadmium.     

Acute effects of Cu on oxygen consumption and 96 hr-LC50 values in the freshwater fish Tilapia sparrmani (Teleostei: Cichlidae) in Mooi River hard water,South Africa

The median lethal copper (Cu) concentration (96 hr-LC₅₀) values for acute Cu toxicity for Tilapia sparrmanii (live mass: 30 ± 8g) in Mooi River hard water of dolomitic origin at 20 °C, pH 7.9, was 68.1 μmol l^?1. At this 96 hr-LC₅₀ value the specific oxygen consumption rate (∈ O₂) decreased by 44.2 (± 2.1) % from a non-exposed value of 6.6 (±0.32) mmol O₂ kg^?1 hr^?1 to 3.63 (±0.23) mmol O₂ kg ^?1 hr^?1. At 46.4 μmol Cu l^?1, 100% of the exposed T. sparrmanii were still alive after 96 hours, but the ∈ O₂ decreased by a mean value of 1.65 (± 0.16) mmol O₂ kg^?1 fish hr^?1 or 25% (± 2.4). Contrary to Pb and Cd, Cu as CuCl₂ 2H₂O was not precipitated in hard water four days after it was dissolved. Thus T. sparrmanii and other cichlids are shown to be more than an order of magnitude more resistant to Cu as a toxicant than most salmonids.     

Effects of sublethal exposure to copper on diel activity of sea catfish,Arius felis

The effect of sublethal exposure to copper on the diel activity of sea catfish, Arius felis, was examined after 72 h static exposure to copper (0.0, 0.05, 0.1, or 0.2 mg l^–1 ). The locomotor behavior of 28 individual fish was electronically monitored in a 16-compartment rosette tank. Activity was quantified by the number of entries into compartments per hour. Controls and copper-exposed fish (0.05 mg l^–1) behaved similarly after treatment and displayed no significant effects of handling or copper exposure on diel activity. However, fish exposed to 0.1 and 0.2 mg l^–1 copper were hyperactive immediately following exposure; these fish had significantly greater numbers of entries into compartments per hour for entire 24-h monitoring periods (P < 0.01) and for the 12 h of photophase (P < 0.001) and of scotophase (P < 0.001) than fish in the other two experimental groups. The normal diel activity cycle was abolished. Additionally, exposure of fish to copper at 0.1 and 0.2 mg l^–1 resulted in significantly less variability in activity (P < 0.05) compared to control fish and those exposed to 0.05 mg Cu l^–1.     

The effect of sub-lethal exposure to copper and the time course of recovery in clean water on biochemical changes in juvenile fish (Acipenser persicus)

This study reports the 96-h LC₅₀ value and tissue copper (Cu) levels and biochemical changes in juvenile fish (Acipenser persicus) exposed to 0.026?mg/l ambient Cu for 1, 7 and 14?days. It then examined the recovery of the same parameters after placing the juvenile fish in clean water for a further period of 28?days. The intestine, kidney and gill Cu levels, plasma glucose, total protein, triglyceride, cortisol, triiodothyronine and thyroxine concentrations, liver protein contents, liver catalase, superoxide dismutase (SOD) and glutathione S-transferase activities were studied. The 96-h LC₅₀ value of Cu was 0.502?mg/l for juvenile A. persicus. The results indicate that Cu exposure produced significant accumulations of Cu in gills and kidney over the treatment time. Sublethal dose of Cu resulted in a short-term increase in plasma glucose, total protein and cortisol levels that decreased with time. After the 28-day recovery phase, there were significant differences in kidney Cu levels and triglyceride concentrations as well as SOD activities between recovery fish treatments and their control groups on day 42. The 28-day recovery phase caused significant decreases in total protein levels and SOD activities of Cu-exposed fish on day 42 compared to day 14. The results suggest that 28?days are insufficient for complete recovery to Cu exposure by juveniles and a longer period would be required for full recovery. Moreover, the study showed that the recovery phase following Cu exposure could change biochemical parameters to levels that are not close to those seen during exposure or control levels.     

Waterborne vs. dietary copper uptake in rainbow trout and the effects of previous waterborne copper exposure

Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to waterborne Cu (22 microg/l) in moderately hard water for up to 28 days. Relative to control fish kept at background Cu levels (2 microg/l), Cu-preexposed fish displayed decreased uptake rates of waterborne Cu via the gills but not of dietary Cu via the gut during 48-h exposures to (64)Cu-radiolabeled water and diet, respectively. At normal dietary and waterborne Cu levels, the uptake rates of dietary Cu into the whole body without the gut were 0.40-0.90 ng. g(-1). h(-1), >10-fold higher than uptake rates of waterborne Cu into the whole body without the gills, which were 0.02-0.07 ng. g(-1). h(-1). Previously Cu-exposed fish showed decreased new Cu accumulation in the gills, liver, and carcass during waterborne (64)Cu exposures and in the liver during dietary (64)Cu exposures. A 3-h gill Cu-binding assay showed downregulation of the putative high-affinity, low-capacity Cu transporters and upregulation of the low-affinity, high-capacity Cu transporters at the gills in Cu-preexposed fish. Exchangeable Cu pools in all the tissues were higher during dietary than during waterborne (64)Cu exposures, and previous Cu exposure reduced waterborne exchangeable Cu pools in gill, liver, and carcass. Overall, these results suggest a quantitatively greater role for the dietary than for the waterborne route of Cu uptake, a key role for the gill in Cu homeostasis, and important roles for the liver and gut in the normal metabolism of Cu in fish.     

Do Larvae and Ovipositing Chironomus riparius (Diptera: Chironomidae) Females Avoid Copper-Contaminated Environments?

Studies on the avoidance behavior of aquatic organisms to contaminants have confirmed that such behavior can be relevant in field situations. However, almost all toxicity tests involve the forced exposure of organisms to toxicants. In particular, despite the importance of Chironomus riparius Meigen larvae in sediment toxicity testing, only a few studies on avoidance behavior have been performed. This study investigated the ability of different life stages of C. riparius, including ovipositing females, first-, second-, and fourth-instar larvae, to avoid copper-contaminated environments. Ovipositing females were given a choice between a control and copper solution (1.3 mg Cu l^{? 1}). First-instar larvae were provided with a choice between a control and a copper (2.0 mg Cu l^{? 1})-spiked sediment. Both second- and fourth-instars were exposed to a copper gradient (0.38–3.4 mg Cu l^{? 1}) in a flow-through system. None of the life stages avoided copper, even though the highest concentrations caused lethal effects on midges. The avoidance behavior of C. riparius is not a sensitive endpoint to assess copper sublethal toxicity.     

The Effects of Acute Waterborne Exposure to Sublethal Concentrations of Molybdenum on the Stress Response in Rainbow Trout,Oncorhynchus mykiss

To determine if molybdenum (Mo) is a chemical stressor, fingerling and juvenile rainbow trout (Oncorhynchus mykiss) were exposed to waterborne sodium molybdate (0, 2, 20, or 1,000 mg l^-1 of Mo) and components of the physiological (plasma cortisol, blood glucose, and hematocrit) and cellular (heat shock protein [hsp] 72, hsp73, and hsp90 in the liver, gills, heart, and erythrocytes and metallothionein [MT] in the liver and gills) stress responses were measured prior to initiation of exposure and at 8, 24, and 96 h. During the acute exposure, plasma cortisol, blood glucose, and hematocrit levels remained unchanged in all treatments. Heat shock protein 72 was not induced as a result of exposure and there were no detectable changes in total hsp70 (72 and 73), hsp90, and MT levels in any of the tissues relative to controls. Both fingerling and juvenile fish responded with similar lack of apparent sensitivity to Mo exposure. These experiments demonstrate that exposure to waterborne Mo of up to 1,000 mg l^-1 did not activate a physiological or cellular stress response in fish. Information from this study suggests that Mo water quality guidelines for the protection of aquatic life are highly protective of freshwater fish, namely rainbow trout.     

Evaluation of lead and chromium tolerance and accumulation level in Gomphrena celosoides: a novel metal accumulator from lead acid battery waste contaminated site in Nigeria

Abstract

Biology, tolerance, and metal (Pb and Cr) accumulating ability of Gomphrena celosoides were studied under hydroponic conditions. The seedlings were raised in Hoagland’s solution containing different concentrations of Pb (0, 500, 1000, 1500, 2000, 3000, 4000, and 5000?mg l^?1) and Cr (0, 50, 100, 150, 200, 300, and 400?mg l^?1). Biomass and metal accumulation in different plant parts were determined at seven (7) and fourteen (14) days after stress. Antioxidant enzyme activities, protein, and proline contents were estimated in stressed and unstressed plants. Gomphrena celosoides was able to tolerate Pb and Cr concentrations up to 4000 and 100?mg l^?1, respectively in hydroponic solution. Metal accumulation was concentration and duration dependent with the highest Pb (21,127.90 and 117,985.29?mg kg^?1) and Cr (3130.85 and 2428.90?mg kg^?1) in shoot and root, respectively found in the plants exposed to 5000?mg l^?1 Pb and 400?mg l^?1 Cr for 14?days. Proline, antioxidant enzyme activities, and protein contents were the highest in plant exposed to higher Pb and Cr concentrations for 7 and 14?days. Gomphrena celosoides could be considered as Pb and Cr accumulator with proline and increase in antioxidant enzyme activities being the tolerance mechanisms.     

Uptake and accumulation of zinc in juvenile rainbow trout,Salmo gairdneri

Synopsis The toxicity of zinc to rainbow trout was determined and the 72 h median lethal concentration was found to be 2.00 mg l^–1 in freshwater, hardness 7.50 mg l^–1 as calcium. An insignificant increase in zinc concentration of internal tissues occurred in fish exposed to 1.52 mg l^–1 in freshwater for 72 h. However, there was a significant uptake of zinc by gills and the body surface. Fish exposed to 10 mg l^–1 zinc for 72 h in two-thirds sea water showed significant zinc uptake by liver, rectum and muscle, when compared to control fish. Drinking rate decreased from 1.43 to 0.26 ml kg^–1 h^–1 when zinc sulphate was added to freshwater. Trout adapted to two-thirds sea water showed no decrease in drinking, about 7 ml kg^–1 h^–1 when zinc was added to the water.     

Heavy metal concentrations in,and human health risk assessment of,three commercially valuable fish species in the lower Niger River,Nigeria

The concentrations of Fe, Mn, Ni, Pb and V in water, sediment and the gill, liver and muscle tissues of Synodontis resupinatus, Heterotis niloticus and Clarias gariepinus, all commercially important fish species of the lower Niger River, were investigated in 2015. Water, sediment and fish samples were collected for six months and heavy metals were determined using an Atomic Absorption Spectrometer. Fe ranked highest in water and sediment, with concentrations of 2.74 mg l^?1 and 61.60 mg kg^?1, respectively. Metals followed the magnitude of Fe > Mn > Ni > V > Pb in the water and Fe > Mn > V > Ni > Pb in the sediments. Metal concentrations were higher in the tissues of S. resupinatus compared with H. niloticus and C. gariepinus. Fe was also highest in the gills, liver and muscle of the three fish species. Its highest concentration of 132.97 mg kg^?1 dry weight was recorded in the gills of S. resupinatus. Bioconcentration factors of metals ranged from 8.79 for Mn in H. niloticus muscle to 67.99 for Ni in S. resupinatus gills. The fish species studied pose no health risk for all metals studied, because the target hazard quotient was less than 1 and the estimated daily intakes of the metals were below the reference doses.     

Recovery of fat snook, Centropomus parallelus (Teleostei: Perciformes) after subchronic exposure to copper

We studied the recovery of juvenile fat snook (Centropomus parallelus) after subchronic exposure to different concentrations of copper. Healthy juveniles (1.98 g) were exposed to 25 or 50 μg Cu/L for 30 days (12 replicates with 5 fish in each one), and recovery was observed at 0, 4, 10, and 30 days after exposure (3 replicates with 5 fish in each one). Copper genotoxicity in exposed individuals was observed using a micronucleus assay, and recovery was not observed even 30 days post-exposure. Copper accumulation was observed in fish exposed to 25 or 50 μg/L of copper in the gills (14.4 and 34.4 μg/g, respectively) and muscle (5.7 and 5.5 μg/g, respectively), and a return to normal copper levels (6.0 μg/g for gills and 2.5 μg/g for muscle) was observed 4 and 30 days post-exposure in the gills and muscle tissues, respectively. Glutathione S-transferase (GST) was 80% inhibited in individuals exposed to copper and returned to normal levels for fish exposed to basal concentrations within 10 days. Although copper accumulation in tissues dispersed 30 days post-exposure, no recovery from genotoxicity was observed during this time. Thirty days was not enough to recover juvenile fat snook following subchronic exposure to copper.     

Cd, Cu, Zn, Se, and Metallothioneins in Two Amphibians, Necturus maculosus (Amphibia, Caudata) and Bufo bufo (Amphibia, Anura)

The accumulation of cadmium, its affinity for metallothioneins (MTs), and its relation to copper, zinc, and selenium were investigated in the experimental mudpuppy Necturus maculosus and the common toad Bufo bufo captured in nature. Specimens of N. maculosus were exposed to waterborne Cd (85???g/L) for up to 40?days. Exposure resulted in tissue-dependent accumulation of Cd in the order kidney, gills > intestine, liver, brain > pancreas, skin, spleen, and gonads. During the 40-day exposure, concentrations increased close to 1???g/g in kidneys and gills (0.64?C0.95 and 0.52?C0.76; n?=?4), whereas the levels stayed below 0.5 in liver (0.14?C0.29; n?=?4) and other organs. Cd exposure was accompanied by an increase of Zn and Cu in kidneys and Zn in skin, while a decrease of Cu was observed in muscles and skin. Cytosol metallothioneins (MTs) were detected as Cu,Zn?Cthioneins in liver and Zn,Cu?Cthioneins in gills and kidney, with the presence of Se in all cases. After exposure, Cd binding to MTs was clearly observed in cytosol of gills as Zn,Cu,Cd?Cthionein and in pellet extract of kidneys as Zn,Cu,Cd?Cthioneins. The results indicate low Cd storage in liver with almost undetectable Cd in liver MT fractions. In field trapped Bufo bufo (spring and autumn animals), Cd levels were followed in four organs and found to be in the order kidney > liver (0.56?C5.0???g/g >0.03?C0.72???g/g; n?=?11, spring and autumn animals), with no detectable Cd in muscle and skin. At the tissue level, high positive correlations between Cd, Cu, and Se were found in liver (all r?>?0.80; ???=?0.05, n?=?5), and between Cd and Se in kidney (r?=?0.76; n?=?5) of autumn animals, possibly connected with the storage of excess elements in biologically inert forms. In the liver of spring animals, having higher tissue level of Cd than autumn ones, part of the Cd was identified as Cu,Zn,Cd?Cthioneins with traces of Se. As both species are special in having liver Cu levels higher than Zn, the observed highly preferential Cd load in kidney seems reasonable. The relatively low Cd found in liver can be attributed to its excretion through bile and its inability to displace Cu from MTs. The associations of selenium observed with Cd and/or Cu (on the tissue and cell level) point to selenium involvement in the detoxification of excessive cadmium and copper through immobilization.     

Embryonic and larval development of brown trout, Salmo trutta L.: exposure to aluminium, copper, lead or zinc in soft, acid water

Freshly fertilized ova, eyed ova and yolk-sac fry of brown trout, Salmo trutta L., were exposed to each of four trace metals (aluminium: 6000 nmol l^?1; copper: 80 nmol l^?1; lead: 50 nmol l^?1; zinc: 300 nmol l^?1) while held in flowing artificial soft-water media maintained at pH 4.5 or 5.6 and [Ca] 20 or 200 μmol l^?1. In continuous exposure from fertilization, survival of ova was severely affected at pH 4.5 and [Ca] 20 μmol l^?1, regardless of the presence of Cu, Pb or Zn; Al reduced embryonic mortality and improved hatching success. High ambient [Ca] at pH 4.5 increased egg survival. At ‘swim-up’, surviving fry exposed to Al or Pb had lower whole body Ca, Na and K content, irrespective of pH or ambient [Ca]. Cu reduced whole body Ca and K content at pH 5.6 and [Ca] 200 μmol^?1, and whole body Ca, Na and K content in the other media. Zn reduced whole body mineral content at pH 5.6 and [Ca] 20 μmol l^?1. Whole body Mg content was reduced by all trace metals at pH 5.6 and [Ca] 20 μmol l^?1, and by Cu at pH 5.6 and [Ca] 200 μmol l^?1. Al and Cu impaired skeletal calcification at pH 5.6 at both ambient [Ca]; Pb only at [Ca] 20 μmol I^?1. Zn enhanced calcification at pH 4.5 and [Ca] 200 μmol l^?1. In the absence of trace metals, low pH reduced body Ca, Na, K content and skeletal calcification at [Ca] 200 μmol l^?1. The uptake of Ca, Na and K, measured at regular intervals from hatching was impaired to the same extent by all treatments at pH 4.5, irrespective of ambient [Ca] or trace metal presence. At pH 5.6, irrespective of ambient [Ca], Al, Cu and Pb impaired Ca and K uptake. The rate of Na uptake was reduced by Al and Cu. Al-treated yolk-sac fry, exposed to low ambient [Ca] from 200–300° days post-hatch, suffered high mortalities regardless of pH. Ca, Na and K uptake was impaired by all treatments at pH 4.5, and by Al and Cu at pH 5.6 in a similar exposure period. The development of the early stages of brown trout in the presence of trace metals is discussed in relation to recruitment failure in areas of soft, acid water.     

Calcium‐dependent behavioural responses to acute copper exposure in Oncorhynchus mykiss

Using rainbow trout Oncorhynchus mykiss, the present study demonstrated that: (1) calcium (Ca) increased the range of copper (Cu) concentrations that O. mykiss avoided; (2) Ca conserved the maintenance of pre‐exposure swimming activity during inescapable acute (10 min) Cu exposure. Data showed that when presented with a choice of Cu‐contaminated water (ranging from 0 to 454 µg Cu l^?1) and uncontaminated water in a choice tank, O. mykiss acclimated and tested at low Ca concentration (3 mg Ca l^?1) avoided the 10 µg Cu l^?1 only. By contrast, O. mykiss acclimated and tested at high Ca concentration (158 mg Ca l^?1) avoided all the Cu concentrations ≥37 µg l^?1. The Cu avoidance was connected with increased spontaneous swimming speed in the Cu‐contaminated water. When subjected to inescapable Cu exposure (35 µg Cu l^?1), O. mykiss acclimated and tested at low Ca concentration reduced their spontaneous swimming speed, whereas no response was observed in O. mykiss acclimated and tested at high Ca concentration. Collectively, the data support the conclusion that in O. mykiss the behavioural responses to acute Cu exposure are Ca‐dependent.     

Exposure to waterborne copper and high temperature induces the formation of reactive oxygen species and causes mortality in the Amazonian fish <Emphasis Type="Italic">Hoplosternum littorale</Emphasis>

Hoplosternum littorale is an Amazon fish that lives in urban areas surrounded by polluted igarapés, where elevated copper concentrations eventually occur. The central goal of this study was to evaluate the associated effects of high temperature and copper contamination on survival time and biochemical responses of the Amazonian fish species H. littorale. We exposed fish to two nominal dissolved copper concentrations (50 and 500 µg l^?1) and combined temperatures of 28 and 34°C. Our findings showed that the combination of these variables affects the survival time of this species. The activity of the biotransformation enzymes ethoxyresorufin-O-deethylase and glutathione-S-transferase showed no alterations in fish within all treatments. The increase of reactive oxygen species and the decrease in potential total antioxidant capacity promoted the imbalance in the antioxidant system. An induction in superoxide dismutase activity occurred in fish exposed to copper concentrations of 50 and 500 µg l^?1 at both temperatures, suggesting liver impairments. Thus, we suggest that H. littorale is sensitive to copper, and this sensitivity is increased further with exposure to high temperatures, particularly in the survival time and reactive oxygen species formation of this fish species.     

Rapid In Vitro Propagation of Orchid Zygopetalum intermedium

The influence of plant growth substances, medium and potting mixture on protocorm development, differentiation, growth and establishment of Zygopetalum intermedium was assessed. Embryo from mature green but unripe capsule cultured on half strength Murashige and Skoog medium containing 1.5 g l^?1 AC with 0.25 mg l^?1 PBZ and 0.1 mg l^?1 of NAA swollen in 59.7 days, followed by formation of globular bodies in 64 days and protocorm development in 70.7 days. Nitsch medium in combination with 0.5 mg l^?1 of BAP and 0.25 mg l^?1 of Triacontanol resulted in shoot and root differentiation and maximum plant growth in vitro. Plantlets with 4–5 well-developed leaves with roots pre hardened in medium supplemented with 0.25 mg l^?1 each of PBZ and Triacontanol transferred to community pots filled with potting mixture of coco peat and tree fern (1:1) resulted in 72.3% survival ex vitro.     

Quantitative histopathology of Oreochromis niloticus gills after copper exposure

In order to test whether histopathological changes of gills demonstrated a good dose–response relationship with water copper levels, juvenile Nile tilapia Oreochromis niloticus, of both sexes and similar mass (36·3 + 7·7 g), were kept in dechlorinated tap water (temperature 25° C, range ±1° C; pH 6·5–7·5; hardness 74·5 mg l^?1 CaCO₃) and exposed to 40 and 400 μg l^?1 of copper. Gill samples were collected after 3, 7, 14 and 21 days. Six major histopathological changes (oedema, lifting, changes in filament epithelium thickness, lamellar fusion, vasodilatation and aneurisms) and three minor ones (proliferation of the lamellar epithelium, necrosis and adjacent lamellar fusion) were found and their prevalence estimated. The extent and severity of each histopathological change were used to develop a severity gradation scale (SGS). Semi‐quantitative analysis of the histopathological changes and measurements of gill copper deposition levels revealed a good dose– and time–response relationship. Oedemas and aneurisms were significantly correlated with acute exposure periods and lamellar fusion with chronic exposure. Epithelial lifting and changes in filament epithelial thickness were seen at lower and higher metal concentrations, respectively. The data also revealed that the SGS profile of each lesion was dependent of gill copper burden.     

Effects of copper sulphate on Tisbe holothuriae Humes (Copepoda) and development of tolerance to copper

A study was made of the toxicity of copper to the marine copepod Tisbe holothuriae Humes and the development of increased tolerance due to acclimation. In sublethal concentrations, copper causes a prolongation of maturation time and a reduction in offspring production. These effects increased proportionally to the copper concentration used, while the respiratory rate (measured at the highest concentration used: 0.008 mg l^–1 Cu) increased. A tendency for higher tolerance due to acclimation was also observed. The copper induced delay in maturation time observed in the F₂ generation, became less pronounced from F₃ onwards. In animals exposed at 0.008 mg l^–1 Cu the maturation time of F₃ was not statistically different from that of untreated animals. The LC50 48 h also presented an increase due to acclimation in animals exposed at 0.004 (F₅) and 0.008 mg l^–1 Cu (F₃ and F₅ generations), but this increase was not statistically significant.